1991-1992 Annual Report - DigitalCommons@USU

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Annual Reports Western Dairy Center

1992

1991-1992 Annual Report 1991-1992 Annual Report

Various Authors

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Western Dairy Foods Research Center

Researching the Western U.S. Dairy Industry's Future

Annual Report 1991-1992

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WESTERN DAIRY FOODS RESEARCH CENTER

ANNUAL REPORT

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TABLE OF CONTENTS

WDFRC Activities Summary .............................................................................. 1

Operational Advisory Committee ....................................................................... 3

Principal Investigators ....................................................................................... 5

WDFRC Budget Activity .................................................................................... 6

Listing of Projects by Account Number. ............................................................. 8

Annual Project Reports by Research Area:

Product Quality and Nutrition Identifying batch of origin of finished cheese

made in continuous processes .................................................................... 15

PI: Lynn Ogden (BYU)

Utilization of acid whey as a substrate for the

production of food grade cellulases ............................................................ 17

PI: Michael Penner (OSU)

Comparative effects of whey protein concentrate

(WPC), lactose, salt, phosphate, and pH on cooked

yield, bind, and acceptability of turkey rolls and

boneless hams ............................................................................................ 18

PI: Daren Cornforth (USU)

Application of Fourier transform infrared technology

to milk and dalry products ........................................................................... 19

PI: Rodney Brown (USU)

Evaluation of iron-protein complexes in iron-fortified

dairy products ............................................................................................. 22

PI: Arthur Mahoney (USU)

Estimation of individual milk proteins and genetic

variants by multicomponent analysis of amino

acid profiles ................................................................................................. 25


• Curd Formation/Cheese Technology

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Improved control of cheese manufacture through vat monitoring ............... 27

PI: Gary Richardson (USU)

Comparison between 40 and 640 lb blocks of uniform cooling of 640 lb

blocks .......................................................................................................... 29

Pis: Conly Hansen (USU) and J. Antonio Torres (OSU)

Ultra-high Temperature Processing Function of whey proteins and lactose in age gelation of ultra-high

temperature sterilized milk concentrate ...................................................... 34

PI: Donald McMahon (USU)

Controlling age gelation of UHT sterilized milk concentrates ...................... 37


Whey Utilization Using whey for improvement of exposed subsoils

and sodic and saline-sodic soils ................................................................. 39

PI: Conly Hansen (USU)

Microbiology of Starter Cultures Cloning the nisin and other genes of lactic streptococci

into Leuconostoc species and amplification of nisin production .................. 41

Pis: Jeffery Broadbent (USU) and William Sandine (OSU)

Purification of a bacteriocin from Pediococcus

pentosaceus and genetic transfer of plasmid-borne determinants ............ 47

PI: Mark Daeschel (OSU)

Growth of Bifidobacteria in milk: Association with Streptococcus thermophilus

and Lactobacillus species as measured by genetic and

enzymatic probes ........................................................................................ 51

PI: William Sandine (OSU)

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• Microbiology of Starter Cultures (continued)

Influence of preadsorbed protein on adhesion of Usteria monocytogenes to

dairy food contact surfaces ......................................................................... 53


Characterization of milk proteolysis by lactococcal

starter culture strains using amino acid analysis ......................................... 55


Prediction and determination of the efficacy of

nisin in dairy foods ...................................................................................... 56


UF/RO Development of a process for production of UF milk retentate powder ...... 60


High yield, low moisture cheese from homogenized

UF milk ........................................................................................................ 62 • PI: Donald McMahon (USU)

Properties of low-fat yogurt manufactured from

ultrafiltered and ultra-high temperature treated milk ................................... 64

PI: Paul Savello (USU)

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APPENDICES

WDFRC Balance Sheet for FY88 to FY92

New Western Center for Dairy Proteins Research and Technology (WCDPRT) Projects 1992-1993 (approved by OAC, July 10, 1992)

Annual Financial Reports by Project

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WDFRC ACTIVITIES SUMMARY

The Western Dairy Foods Research Center (WDFRC) was very active during its operating year 1991-1992. These activities included:

• continuation of important dairy products/processing research projects;

• sponsoring lectureships at the American Dairy Science Association (ADSA) Annual Meeting held on the Utah State University campus in August, 1991;

• holding the WDFRC Annual Meeting in August, 1991 in coordination with the ADSA meeting;

• completing the review and extension process by the NDPRB;

• communicating to the dairy industry the emerging technologies and research results;

• reporting many project results of WDFRC projects at scientific meetings;

• Technical Advisory Committee (TAC) review of newly proposed research projects.

Twenty (20) research projects were active during this past year. Some research projects came to completion during this time. The five (5) research areas of the WDFRC had active projects. Many projects were actively continuing into the next operating year (1992-1993).

The WDFRC Annual Meeting was held on August 10, 1991 on the Utah State University campus. This meeting was coordinated with the ADSA's Annual Meeting, which was held on this same campus on August 12 to 15, 1991. The Operational Advisory Committee (OAC) of the WDFRC provided very significant input into the future direction and focus of the WDFRC. At this OAC meeting, update report of all WDFRC-sponsored projects were presented.

The Center obtained approval from the NDPRB for a four-year extension of its activities. This extension starts July 1, 1992 and will carry the Center forward to 1996. All supporters, researchers, and staff of the Center are proud of the work the Center has undertaken during its first five-year period. Everyone associated with the Center expects that research and transfer of results will continue to grow on the solid foundation that had been set during the initial years of Center operation.

A saddened event took place during the past year at the WDFRC. Dr. Arthur Mahoney passed away after a bout with cancer and other complications. Dr. Mahoney spearheaded and maintained significant progress in his Center-sponsored and NDPRB directed research involving the iron fortification of cheese. Phase I of this large research effort is complete. We are sure that Dr. Mahoney would have

• been most pleased with the progress and results of this project.

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The TAC reviewed sixteen (16) new research proposals during April and May, 1991 . Nine (9) proposals were eventually accepted by the Director and the OAC. This year the TAC was made up of three (3) dairy industry representatives, one (1) dairy researcher on the faculty of Oregon State University, one (1) NDPRB staff member, one (1) dairy researcher on the faculty of the University of Minnesota, and the WDFRC Director. The Director believes that exposing dairy researchers at other institutions (particularly at other NDPRB Dairy Centers) to the research ideas of WDFRC researchers can lead to meaningful collaborative efforts between Centers. Such collaboration among scientists can prove most beneficial to produce important and timely research results. It is fully expected that having another dairy research from another Dairy Center on the WDFRC TAC will prove to be an excellent vehicle to have researchers "hitchhike" and "brainstorm" new research ideas as well to pool expertise and equipment.

Researchers at the WDFRC were very successful in winning NDPRB Competitive Program research grants during 1992. Five (5) research projects were funded under the competitive program. Researchers at the WDFRC are also very active in pursuing (and winning!) research grants from other sources. Investigators at Utah State University continue to administer a State of Utah Center of Excellence in Dairy Foods Technology. The combined resources of these Centers prove to be an excellent leverage so that more research can be conducted, with results being transmitted/transferred to all supportive and contributing parties .

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OPERATIONAL ADVISORY COMMITTEE 1991-1992

Pursuant to the WDFRC proposal and contract with the National Dairy Promotion and Research Board, the voting members of the Operational Advisory Committee are:

Barbara Blakistone National Dairy Promotion and

Research Board 2111 Wilson Blvd., Suite 600 Arlington VA 22201 (703) 528-4800

Rodney J. Brown, Dean College of Agriculture Utah State University Logan UT 84322-4800 (801) 750-2215

Daniel F. Farkas, Head Department of Food Science

and Technology Wiegand Hall 240 Oregon State University Corvallis OR 97331 (503) 737-3131

Jeff Strandholm Kraft, Inc. Research and Development Div. 801 Waukegan Road Glenview IL 60025 (312) 998-3717

Raj G. Narasimmon Schreiber Foods, Inc. P.O. Box 19010 Green Bay WI 54305 (414) 437-7601

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Sheldon Pratt Oregon Dairy Products Commission 10505 S.W. Barbur Blvd. Portland OR 97219 (503) 229-5033

Paul A. Savello, Director Western Dairy Foods Res. Center Department Nutr. & Food Sciences Utah State University Logan UT 84322-8700 (801) 750-21 06

Harry Papageorge 1616 Farr West Drive Ogden UT (801) 782-9383

Douglas Willrett Marschall Products P.O. Box 592 Madison Wl53701 (608) 276-3600

Boyd Gardner Utah Dairy Commission 8460 North Highway 69 Honeyville UT 84314 (801) 279-8320

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OPERATIONAL ADVISORY COMMITTEE 1991-1992

Gale Moser United Dairymen of Idaho 1864 South Hulls Crossing Preston ID 83263 (208) 852-0560

Tom Holzinger Borden, Inc. 960 Kingsmill Parkway Columbus OH 43229 (614) 431-6623

(continued)

Donald McMahon, Acting Head Department of Nutr. & Food Sciences Utah State University Logan UT 84322-8700 (801) 750-2102

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PRINCIPAL INVESTIGATORS

Floyd W. Bodyfelt Mina R. McDaniel Dept. of Food Science & Technology Dept. of Food Science & Technology Oregon State University Oregon State University Corvallis OR 97331 Corvallis OR 97331

Rodney J. Brown Joseph McGuire Dept. of Nutrition & Food Sciences Dept. of Food Science & Technology Utah State University Oregon State University Logan UT 84322-8700 Corvallis OR 97331

Daren P. Cornforth Donald J. McMahon Dept. of Nutrition & Food Sciences Dept. of Nutrition & Food Sciences Utah State University Utah State University Logan UT 84322-8700 Logan UT 84322-8700

Mark A. Daeschel Lynn V. Ogden Dept. of Food Science & Technology Dept. of Food Science & Nutrition Oregon State University Brigham Young University Corvallis OR 97331 Provo UT 84602

• Bruce L. Geller Michael H. Penner Dept. of Microbiology Dept. of Food Science & Technology Oregon State University Oregon State University Corvallis OR 97331 Corvallis OR 97331

Conly L. Hansen Gary H. Richardson Dept. of Nutrition & Food Sciences Dept. of Nutrition & Food Sciences Utah State University Utah State University Logan UT 84322-8700 Logan UT 84322-8700

Jeff Broadbent William E. Sandine Dept. of Nutrition & Food Sciences Dept. of Microbiology Utah State University Oregon State University Logan UT 84322-8700 Corvallis OR 97331

Arthur W. Mahoney Paul A. Savello Dept. of Nutrition & Food Sciences Dept. of Nutrition & Food Sciences Utah State University Utah State University Logan UT 84322-8700 Logan UT 84322-8700

J. Antonio Torres Janine E. Trempy Dept. of Food Science & Technology Dept. of Microbiology Oregon State University Oregon State University

• Corvallis OR 97331 Corvallis OR 97331

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• WDFRC BUDGET ACTIVITY 1991-1992 FISCAL YEAR

INSTITUTIONAL SUPPORT Utah State University

4.2 FTE Dairy Research $201,000 Facilities* 250,000 20% indirect (NDPRB-permitted) 90,200

Oregon State University 0.95 FTE Dairy Research 59,000 Facilities* 230,000 20% indirect (NDPRB-permitted) 57,800

Brigham Young University 0.4 FTE Dairy Research 25,000 Facilities* 15,500 Scholarships/Research Assistantships 14,500 Direct Research Support 4,400 20% indirect (NDPRB-permitted) 11,780

TOTAL INSTITUTIONAL SUPPORT $959,180

• RESEARCH CONTRIBUTIONS:

NATIONAL DAIRY PROMOTION AND RESEARCH BOARD 400,000

REGIONAUINDUSTRY SUPPORT:

Utah Dairy Commission 50,000 United Dairymen of Idaho 50,000 Oregon Dairy Products Commission 40,000 Western Dairy Farmers' Promotion Assoc. 10,000

Kraft-General Foods, Inc. 5,000 Schreiber Foods, Inc. 5,000 Marschai-Rhone Poulenc, Inc. 5,000 Borden, Inc. 5,000 Western Dairymen Cooperative, Inc. 2,500

USDA-ARS 200,000

State of Utah Center of Excellence "Center for Dairy Foods Technology" 130,000

SUBTOTAL REGIONAUINDUSTRY SUPPORT 502,500

• FY92 TOTAL DAIRY RESEARCH CONTRIBUTIONS $902,500

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WDFRC BUDGET ACTIVITY 1991-1992 FISCAL YEAR

(continued)

FY91 BALANCE FORWARD

TOTAL AVAILABLE FUNDS FOR FY92 RESEARCH

FY92 COMMITTED FUNDS Western Dairy Foods Research Center USDA-ARS Support State of Utah Center of Excellence WDFRC Administrative

TOTAL FY92 COMMITTED FUNDS


* Estimated values

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(424,886) (200,000) (130,000) (50,000)

$41,616

$944,116

($804,886)

$139,230

• • • Western Dairy Center Listing of Funded Projects (Allocated) By Account Number

Project Funding Allocated Thru FY 1996

Project Project Account Proj Principal Research Research Start Ending Number Numb Investigator Area Title Date Date FY88 FY89 FY90 FY91 EY..22 FY93 FY94 FY95 FY96

547766 189 Hansen, C.L. Ultrafiltration Cogeneration of Biogas and 07/01/87 06/30/89 29589 29589 0 0 0 0 0 0 0 and Reverse Single Cell protein From Osmosis Ultrafiltration Permeate

and Whey

547767 188 Broadbent, J. Microbiology Cloning the Nisin and Other 07/01/87 06/30/93 41680 34440 32105 33711 35396

of Starter Genes of Lactic Cultures Streptococci into

Leuconostoc Species and Amplification of Nisin Production

547768 182 Mahoney, A.W. Product Quality Iron fortification of cheese 08/01/87 06/30/90 20944 21753 0 0 0 0 0 0 0 curd

547769 190 Richardson, G.H. Product Quality Acquisiton of Zymark II 07/01/87 06/30/90 5000 0 0 0 0 0 0 0 0 Robot for Laboratory Automation Studies

547778 191 General Expenses-NDPRB 0 0 27 0 0 0 0 0 0

547779 191 General Expenses-Non-NDPRB 0 0 0 67529 0 0 0 0 0

547780 181 Richardson, G .H. Curd Improving Yield and 07/01/87 06/30/91 22620 22620 22620 0 0 0 0 0 0

Formation/ Physical Properties of Cheese Tech. Mozzarella Cheese

547781 183 Ernstrom, C.A. Ultrafiltration Continuous Production of 09/01/87 08/31/89 8700 0 0 0 0 0 0 0 0 and Reverse Cottage Cheese From Osmosis Ultra-filtrated skim milk

Retenate

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• • • Project Funding Allocated Thru FY 1996

Project Project Account Proj Principal Research Research Start Ending Number Numb Investigator Area Title Date Date FY88 FY89 FY90 FY91 FY92 FY93 FY94 FY95 FY96

547782 184 Cornforth, D.P. Product Quality Evaluation of Milk Proteins 07/01/87 6/30/90 15327 16512 0 0 0 0 0 0 0 as Whitening Agents in Processed Meats and Poultry Products

547783 186 Olsen, R.L. Cuid Interaction of Protein and 07/01/87 06/30/89 21750 21410 0 0 0 0 0 0 0 Formation/ Polysaccharides in Chymosin Cheese Tech. and Acid Coagulation of Milk

547784 187 Richardson, G.H. Cuid Improved Control of Cheese 07/01/87 04/30/92 35300 14000 14000 0 0 0 0 0 0 Formation/ Manufacture Through Vat Cheese Tech. Monitoring

547785 191 Savello, P.A. Administrative Western Dairy Foods 07/01/87 06/30/92 20000 30000 50000 50000 50000 50000 50000 50000 50000 Expenses Research Center

Administrative Account

547802 192 Ernstrom, C.A. Cuid Effect of Milk Clotting 07/01/87 06/30/88 37660 0 0 0 0 0 0 0 0 Formation/ Enzymes on the Curing and Cheese Tech. Quality of Cheddar Cheese

547804 188 Sandine, W .E. Microbiology Cloning the Nisin and Other 07/01/87 06/30/93 44565 43033 45274 47537 49914 0 0 0 0 of Starter Genes of Lactic Cultures Streptococci into


547810 194 Sandine, W .E. Microbiology Characterization of 07/01/87 06/30/90 18110 18573 18573 0 0 0 0 0 0 of Starter Bacteriophage Receptor Cultures Sites of Lactic

Streptococci

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547811 195 Bodyfelt, F.W. Product Quality Rapid Assay for Heat 08/01/87 12/31/90 18285 3035 15260 0 0 0 0 0 0

Resistant Microbial Proteases in Raw Milk by a Simple Casein Denaturation Method

547812 196 Bodyfelt, F.W. Microbiology Production of Omega-3 Fatty 09/01/87 12/31/90 24350 5415 24890 0* 0 0 0 0 0

of Starter Acids by Genetically Cultures Altered Fungi and Lactic

Acid Bacteria

547813 197 McGuire, J. Product Quality Characterization of the 01/01/88 12/31/90 20910 31043 24643 0 0 0 0 0 0 Post-Absorbtive Behavior of B-Lactoglobulin For Control of Spore and Microbial Adhesion

547814 198 Sandine, W .E. Microbiology Studies on the Growth and 07/01/87 06/30/89 18110 18573 0 0 0 0 0 0 0 of Starter Survival of Bifidobacterium Cultures Species in Milk

547822 183 Ogden, L.V. Ultrafiltration Continuous Production of 09/01/87 08/31/89 0 8200 0 0 0 0 0 0 0

and Reverse Cottage Cheese From Osmosis Ultrafiltrated Skim Milk

Retenate

547823 199 Ogden, L.V. Product Quality Method for Identifying 07/01/88 06/30/90 0 6100 6100 0 0 0 0 0 0 Batch of Origin of Semi-continuous Cheese Processes

547825 201 Brown, R.J. Product Quality Application of Fourier 07/01/88 12/31/91 0 73350 25205 26366 0 0 0 0 0

Transform Infrared Technology to Milk and Dairy Products

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547827 267 Brown, R.I. Product Quality Estimation of Individual 07/01/88 12/31/92 0 145425 50594 51874 0 0 0 0 0

Milk Proteins and Genetic Variants by Multicomponent Analysis of Amino Acid Profiles

547828 204 Savello, P.A. Ultrafiltration Properties of Low-fat Yogurt 08/01/88 09/30/91 0 28500 29700 0 0 0 0 0 0

and Reverse Manufactured from Ultra-Osmosis filtered and Ultra-high Temp-

Treated Milk

547833 203 Daeschel, M.A. Microbiology Purification of a 07/01/88 12/31/91 0 16062 18100 20300 0 0 0 0 0

of Starter Bacteriocin From Cultures Pediococcus Pentosaceus and

Genetic Transfer of the Plasmid Borne Determinant

547834 202 Torres, J.A. Curd Cheddar Cheese Blocks: 07/01/88 12/31/90 0 26530 16212 0 0 0 0 0 0

Formation/ Effect of Cheese Composition Cheese Tech. and Cooling Method

547838 206 Hansen, C.L. Curd Comparison Between 40 and 11/01/88 12/31/91 0 2300 11000 4600 0 0 0 0 0 Formation/ 640 lb Blocks of Uniform Cheese Tech. Cooling of 640 lb Blocks

547839 200 McMahon, D.J. Ultrafiltration Variations in Casein 09/01/88 06/30/92 0 24936 26543 27877 0 0 0 0 0

and Reverse Composition of Milk High Osmosis Yield, Low Moisture Cheese

From Homogenized Milk

547841 208 Mahoney, A.W. Product Quality Evaluation of Iron-Protein 10/04/88 12/31/92 0 35505 l8o9oll38442ll41422l 0 0 0 0

Complexes in Iron-Fortified Dairy Products


Project Project Account Proj Principal Research Research Start Ending Number Numb Investigator Area Title Date Date FY88 FY89 EY..2Q EY..2.l. FY92 FY93 FY94 FY95 FY96

547848 210 Hansen, C.L. Curd A New Method for Measuring 03/01/89 03/01/91 0 23912 0 0 0 0 0 0 0 Formation/ Syneresis of Renneted Gels Cheese Tech. Applied to Development of

Cheese

547862 206 Torres, J.A. Curd Cooling Rate of Cheddar 11/01/88 12/31/91 0 11549 18320 2475 0 0 0 0 0 Formation/ Cheese: Comparison Between Cheese Tech. 40 and 640 lb Blocks of

Uniform Cooling of 640 lb Blocks

547863 209 McDaniel, M.R. Product Quality Optimization of the Sensory 07/01/88 06/30/90 0 32127 0 0 0 0 0 0 0 Qualities of Flavored Yogurt

547864 207 Daeschel, M.A. Microbiology Prediction and 07/01/88 QJlU/9.7 0 18230 21177 0 182401 0 0 0 0 of Starter Determination of the Cultures Efficacy of Nisin in Dairy

Foods

547867 211 McMahon, D.J. UHf Function of Whey Proteins 07/01/89 06/30/92 0 0 25000 25000 0 0 0 0 0 Processing and Lactose in Age Gelation

of Ultra-High Temperature Sterilized Milk Concentrate

547877 212 Torres, J.A. Product Quality Acid Whey Utilization: 07/01/89 06/30/91 0 0 l2o58ol[22797] 0 0 0 0 0 Functional Properties of a Food Grade Stabilizer Produced by Lactobacillus Plantarum from Acid Whey

547898 204 Savello, P.A. UF/RO Membrane fractionation of 07/01/90 06/30/91 0 0 0 33500 0 0 0 0 0 immunoglobulins from milk and whey

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• Account Proj Principal Number Numb Investigator

547899 210 Hansen, C.L.

547900 211 McMahon, D.J.

Research ~

UF/RO

UHf Processing

547901 181 Richardson, G.H. Product Quality

547902 198 Sandine, W.E. Microbiology

of Starter Cultures

547903 212 Penner, M. Product Quality

547910 243 Brown, R.I. Microbiology

of Starter Cultures

547936 183 Ogden, L.V. Product Quality

Research Title

Development of a process

for production of UF milk retentate powder

Controlling age gelation of

UHTsterilizedmilk concentrates

Causes and prevention of sticky texture in Mozzarella cheese

Growth ofbifidobacteria in

milk: Association with Streptococcus thermophilus and Lactobacillus species as measured by genetic and enzymatic probes

Utilization of acid whey as

a substrate for the production of food grade cellulases

Characterization of milk

proteolysis by lactococcal starter culture strains using amino acid analysis

Identifying Batch of Origin

of Finished Cheese Made in Continuous Processes

• Project Start Date

07/01/90

07/01/90

07/01/90

07/01/90

07/01/90

07/01/90

07/01/91

Project Ending Date

06/30/93

06/30/93

06/30/91

12/31/92

06/30/92

12/31/92

12/31/92

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• Project Funding Allocated Thru FY 1996

0 0 0 61405 64415 0 0 0 0

0 0 0 34440 38740 0 0 0 0

0 0 0 19380 0 0 0 0 0

0 0 0 1254531 26315 0 0 0 0

0 0 0 20583 0 0 0 0 0

0 0 0 22000 23100 0 0 0 0

0 0 0 0 12000 0 0 0 0



547950 241 Cornforth, D.P. Product Quality Comparative Effects of Whey 07/01/91 12/31/92 0 0 0 0 23990 0 0 0 0

Protein Concentrate (WPC), Lactose, Salt, Phosphate, and pH on Cooked Yield, Bind, and Acceptability of Turkey Rolls and Boneless Hams

547951 203 Daeschel, M. Microbiology Influence of Preadsorbed 07/01/91 06/30/94 0 0 0 0 30354 28399* 29544 0 0

of Starter Protein on Adhesion of Cultures Listeria monocytogenes

to Dairy Food Contact Surfaces

547952 263 Hansen, C.L. Whey Utilization Using Whey for Improve- 07/01/91 06/30/93 0 0 0 0 21000 14000 0 0 0 ment of Exposed Subsoils and Sorlie and Saline-Sodic Soils

***'lUI' AL AI.1.1)CATED FUNDS*** 402900 762722 524013 1635269 4248861 .22122 79544 50000 50000

lEGEND:

D - Increase in project funding approved for FY s

• - Decrease in project funding approved for FY s

L_2 - Extension (no cost) approved

!.. .... .)- Project revision approved

L_) - Projected funding 14

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Project Title:

Identifying Batch of Origin of Finished Cheese Made in Continuous Processes.

Period:

July 1991 to May 1992

Personnel:

L. V. Ogden, Associate Professor, Food Science and Nutrition, Brigham Young University; I. L. Holt, Associate Professor, Manufacturing Engineering and Technology, Brigham Young University; Salvador Uy Parco, Graduate Assistant, Food Science and Nutrition, Brigham Young University.

Funding Sources:

Western Dairy Foods Research Center, non-dairy board monies; Brigham Young University Department of Food Science and Nutrition .

. Objectives:

To develop a system to track cheese batches through pressing towers so that blocks exiting the tower can be reliably marked with its batch of origin .

Results:

On-line colorimetry was used to distinguish between alternately color-marked batches of cheddar cheese. Marker levels were selected that would minimize visual difference while providing sufficient instrumental discrimination between marked and unmarked batches. Control cheese contained 53.2 Jtl of double-strength annatto per kg of milk. Marked cheese contained an additional 6.6 Jtl of annatto per kg of milk. This is a level that was determined to be just noticeable visually in earlier work. A colorimeter was used to record color values of cheese blocks exiting the tower. A rolling t-test was used to compare color readings of 10 preceding blocks with 10 following blocks as each block was ejected from the tower. It was expected that the absolute value of the t-test statistic would be maximum when comparing samples that are in different batches. Therefore, the absolute value of the t statistic should be maximum at the seam between batches. In laboratory trial 81% of the seams were correctly identified. In two separate plant trials 74% of the seams were correctly identified. Similar experiments with tumeric as a marker did not produce better results.

Since 100% correct seam identification is necessary for the marker concept to work and since greater batch color differences are not expected to be acceptable, the color marker idea was shelved in favor of the following method of identification. It should

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be possible to identify the batch seam if the amount of cheese in the tower is known between batches. The seam between batches would correspond with the ejection of that much cheese. An ultrasonic sensor mounted at the top of the tower to measure the headspace distance between batches is being tested. Microwave and laser sensors are being investigated. Such sensors could not only track batches but also control cheese levels in the towers, a capability that does not now exist. A prototype system to provide both batch identification and tower level control is being designed.

Impact of Research

Inability to accurately identify the batch of origin of specific units of cheese for recall or downgrading is a problem in plants using cheddaring and/or block forming towers in cheese manufacturing. One plant manager estimated the unnecessary value reduction on his cheese to be $250,000 per year assuming that 6.5 million pounds of cheese was unnecessarily downgraded and an average reduction in value of $.04 per pound. Ability to identify batch of origin would prevent that loss .

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WDFRC Progress Report. 1992

Project Title: Utilization of Acid Whey as a Substrate for the Production of Food Grade Cellulases.

Personnel: M. H. Penner, Assistant Professor, Dept. Food Science and Technology, Oregon State University.

Funding: Western Dairy Foods Research Center

Objectives:

A. To demonstrate the feasibility of utilizing acid whey as the primary substrate for cellulase enzyme production by the mold Trichoderma reesei.

B. To demonstrate that acid whey has beneficial properties that make it an attractive substrate for enzyme production by acid tolerant fungi in general.

Results: Experiments with Trichoderma reesei growing on whey-based media show that the start of net protein production is earlier when using whey permeate as a substrate than when using pure whey. Protein production rates on the two whey-based substrates are approximately the same. Both whey and whey permeate provide sufficient nitrogen for growth and apparent maximum enzyme production. In pure whey-based media, a higher initial pH gives an earlier start of enzyme production. Size exclusion chromatography has been used to partially purify the enzyme fraction low molecular weight components, principally lactose and peptides. There is a lag period of 3-4 days prior to the start of protein/ enzyme secretion when Trichoderma is cultured on either of the whey substrates. This is longer than that observed on a cellulose-based medium. Maximum enzyme production occurs after approximately 10% of available lactose (inducer) is consumed from the whey-based media. Addition of 0.1% cellulose (solkafloc SW 40) has not significantly improved the induction.

Impact of Research: The research has demonstrated that Trichodenna is capable of growth and enzyme production on whey-based media. Quantification of enzyme yields and cost parameters are still being determined. The findings, when completed, will be published in an industry-oriented journal, such as the Journal of Industrial Microbiology. We anticipate that our publication will lead several biotechnology-oriented companies to consider using whey-based media in their fermentation/protein production systems.

Publications: A manuscript detailing this material will be submitted following our complete data analysis. Preliminary reports of. our studies may be obtained from M.H. Penner .

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Comparative Effects of Whey Protein Concentrate, Lactose, Salt, Phosphate and pH on Cooked Yield, Bind and Acceptability of Turkey

Rolls and Boneless Hams

Progress Report - June 1, 1992

Daren Cornforth Nutrition and Food Sciences

Utah State University

Phosphates are widely used in cooked meats to increase cooked yields and bind. The primary effect of phosphates is to increase meat pH, thereby increasing the water holding capacity of the muscle. However, phosphates are relatively expensive, and nutritional concerns have been expressed concerning the role of phosphates in decreasing absorption of calcium. The purpose of this work is to determine the feasibility of using high pH-whey protein concentrate, rather than phosphate, to increase the cooked yield of turkey rolls and ham. Preliminary work here has shown that 3% whey protein concentrate (WPC) increased the cooked yield of turkey rolls by about 2 % (from 95 to 97%), in the absence of phosphate. The appatatus of Dargan and Savello (1991) used for measurement of breaking strength of protein films has been adapted for measurement of bind strength of 1 em slices of turkey rolls. A CUI Advantage 386 computer has been purchased (with State of Utah funds), and is currently being programmed to rapidly collect and graph bind-strength of turkey rolls. (Bind strength is measured as the force in grams required to force a 2 em diameter rod through the center of a lcm thick meat slice. Data is collected continuously over the 2 minute period required to penetrate the slice, resulting in over 500 data points per graph; hence the need for the computer). The effects of pH adjustment (with phosphate or sodium hydroxide) on bind strength of turkey rolls is currently underway. A 6 month extension has been requested for completion of the project .

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Project Title: Application of Fourier Transform Infrared Technology to Infrared Technology to Milk and Dairy Products

Personnel: Rodney J. Brown, Dept. of Nutrition and Food Sciences, Utah State University.

lyan v. Mendenhall, Dept. of Nutrition and Food Sciences, Utah State University.

Steve Stembridge, Dept. of Nutrition and Food Sciences, Utah State University.

Funding: Western Dairy Foods Research Center

Objectives:

1.

2.

3.

4 .

5.

6.

Find a set of wavelengths in the infrared spectrum that respond to changes in fat, protein, and lactose concentrations.

Find a set of wavelengths in the infrared spectrum that do not respond to changes in saturation level, chain length, and level of free fatty acids.

_ Combine (1) and (2) to make a robust set of wavelengths common to all constraints.

Determine the individual spectra and common wavelengths to milk fat, protein and lactose.

Find a set of wavelengths common to the milk components (4) and to the robust set (3).

Statistically calibrate for testing samples of unknown composition using only this set of wavelengths (9) and milk samples chemically tested for fat, protein, lactose, and moisture. (Less than 1600 cm-1 wavenumber should be used if possible.)

7. Calibrate, using wavenumbers greater than 2700 cm-1, to determine saturation level of fat in dairy products (especially cheese).

Results:

To identify wavelengths which respond to changes in fat concentration, we prepared a series of milk samples where the fat level varied and all other components were held constant. The correlation coefcient for absorbance and fat concentration was calculated at each wavelength. A similar experiment was performed to determine wavelengths that linearly respond to changes in protein concentration. To determine wavelengths that linearly respond to changes in lactose concentration, a series of lactose solutions were prepared where the lactose concentration varied from 1 to 5.5%. We then selected wavelengths which were responsive to fat, protein, and lactose concentrations, and

• relatively unaffected by saturation, chain-length, and lipolysis. Nine calibration

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• standards (each composed of milk from a separate herd) were purchased. The FTIR was calibrated using these standards and Partial Least Squares (PLS) statistics. The concentration of fat, protein, and lactose in these samples was then predicted using the generated calibration equations. The standard deviations of difference between chemical and predicted values were below the AOAC recommended SD of .06%. This data was obtained with no homogenization or temperature control. A liquid ATR cell was used and 64 scans at 4 cm-1 resolution were averaged to obtain each sample spectrum. Addition of whey powder to non-fat dry milk and non-dairy fat to cheese were quickly and easily detected using FTIR. We are now comparing this method of infrared milk analysis with that used in commercial instruments.

Impact of Research:

This project was requested by one of the parties (Utah Dairy Commission) contributing to the funding of the Western Dairy Foods Research Center. They wanted a fast method for detecting non-dairy products, particularly fat, in products labeled or sold as dairy products. Their justication is that adulterated products now being sold as dairy products will be replaced by real dairy products if it is widely known that such a test exists. In accomplishing this end. we also provided an improved method for measuring fat, protein, lactose and water content in all dairy products. This should assist the dairy industry as use of milk components in all foods becomes a larger share of total milk utilization.

Publication:

• Mendenhall, I. V., R.J. Brown and R. Grappin. 1990. Alternative wavelengths for infrared

measurement of fat, protein, and lactose in milk. 85th American Dairy Science Association Meeting J. Dairy Sci. 73:Supp. 1, 92.

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Richardson, G. H., R. J. Brown, T.C J. Yuan and I.V. Mendenhall, 1990, Combining FTIR, conductance and reflective colorimetry for simultaneous estimation of milk components, abnormal milk, microbial and antibiotic contents in raw milk. XXIII International Dairy Congress. Montreal, Canada.

Mendenhall, I. V. and R. J. Brown, 1991, Spectroscopic detection of whey protein powder in non-fat dry milk. Amer. Chern. Soc. Annual Meeting, Atlanta, GA 201 :4-PAGFD.

Mendenhall, I. V. and R. J. Brown, 1991, Fourier transform infrared determination of whey powder in non-fat dry milk. J. Dairy Sci. 74(9):2896-2900.

Mendenhall, I. V., and R. J. Brown, 1991, Spectroscopic determination of whey powder in non-fat dry milk. 86th American Dairy Science Association Meeting J. Dairy Sci. 74:Supp. 1, 93.

Mendenhall, I. V. 1991, Rapid Determination of Milk Components and Detection of Adulteration using Fourier Transform Infrared Technology. Ph.D. Dissertation, Utah State University, Logan.

Brown, Rodney J., 1989, Infrared Analysis of Biological Substances. Patent in Process .

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Mendenhall, I. V. and R. J. Brown, 1991, Spectroscopic determination of a non-dairy fat in process cheese. In Preparation .

Mendenhall, I. V. and R. J. Brown, 1991, Robust infrared wavelengths for the determination of fat, protein, and lactose in milk. In Preparation .

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• Project Title:

Personnel:

Funding:

Objectives:

Evaluation of Iron-Protein Complexes in Iron-Fortified Dairy Products

Dr. Arthur W. Mahoney, Department of Nutrition and Food Sciences, Utah State University

Dr. Mohan I. Reddy, Dept. of Nutrition and Food Sciences, Utah State University.


The objective of this study is to determine the chemistry of iron-protein complexes in milk as related to cheese making. The mechanism and thermodynamics of iron-protein complex formation is necessary to understand the functional and stability characteristics of iron-milk protein complexes. Elucidation of the mechanism would provide the information to enable the dairy processor to use conditions and/or competitive reagents to maximize complex formation. We have reported earlier (i) the binding of Fe(lll) to different casein and whey protein fractions at pH 6.60, (ii) iron-induced conformational changes in proteins at pH 6.60, and (iii) effect of iron on chymosin hydrolysis and rennet clotting time of milk. In this report we present the effect of pH, ionic strength, and

• dephosphorylation on the binding of Fe(lll) to a 51-casein.

•

Results:

Effect of pH, Ionic Strength and Dephosphorylation on the Binding of Fe(lll) to a 8 1-Casein: Binding of Fe(lll) to a 51-casein was studied as a function of pH (5.6, 6.1, 6.6, 7.2, and 7.8), ionic strength (0.1 and 0.5), and dephosphorylation of protein using diafiltration method and the binding data analyzed by Scatchard equation. pH and ionic strength had no influence on the number of iron binding sites on the protein, which remained constant at n=20. However, binding affinity of Fe(lll) to protein decreased with an increase in pH from 5.6 to 7.8 and ionic strength from 0.1 to 0.5. Thus, from the practical point of view, the binding affinity of Fe(lll) increases as the pH of milk is lowered by microbial action during cheese making. Dephosphorylation of a51-casein decreased the number of iron binding sites on the protein (n=9) indicating that phosphoserine groups play a major role in the binding. In another experiment, partial displacement of protein-bound Ca(ll) by Fe( Ill) was observed for a51-casein. This indicates that Fe( Ill) successfully competes for some of the Ca(ll) binding sites on the protein. Free energy change (flG = -RT In K) calculated for the binding of Fe(lll) to a 51-casein under different pH and ionic strength conditions was negative and low in magnitude (-3.79 and -7.28 k 'Cal M-1 ) indicating that the binding of Fe(lll) to protein is instantaneous and thermodynamically favorable .

Visible-Difference Spectra of Fe(lll) - a 8 1-Casein Complexes: Difference absorption spectra of Fe(lll)-protein complexes in the visible region (350 to 650 nm) were

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carried out to determine the possible amino acid side chain groups involved in the binding of Fe(lll) to a51 -casein at different pHs (5.6 to 7.8). Negative absorption band at 420 nm, and positive absorption bands at 470, 490 and 560 nm were observed for the protein at all pHs. The model spectra of different amino acid-Fe(lll) complexes revealed that the absorption bands at 420 and 565 nm were due to phosphorylserine-Fe(lll) and carboxyi-Fe(lll) complexes, respectively, where as the bands at 470 and 490 nm were possibly due to a chelate site involving carboxyl, nitrogen and oxygen groups. The magnitude of 420 nm band decreased, where as that of 470, 490, and 565 nm bands increased with an increase in pH. This may indicate that the involvement of carboxyl groups in the complex formation increases as the pH is moved away from their isoelectric pH. Thus, phosphorylserines and carboxyl groups of Asp and Glu seem to play a major role in the binding of Fe(lll) by a51-casein.

Fe(lll)-lnduced Conformational Changes in a 5 1-Casein: Conformational changes in a51-casein, especially changes in the environment of aromatic side chains due to binding of Fe(lll) were monitored by following UV-difference spectra and fluorescence emission spectra. The UV-difference spectra of a51-casein induced by Fe(lll) at pH 5.6, 6.1, 6.6, 7.2, and 7.8 had absorption bands at 310 nm indicating the possible involvement of tryptophan residues in charge-transfer type complex formation with Fe(lll). Since it is characteristic of charge-transfer type complexes to quench tryptophan fluorescence in proteins, the accessibility of tryptophanyl residues in as1-casein in the absence and presence of Fe(lll) was followed by iodide quenching at pH 6.6. Intrinsic tryptophan fluorescence of a51-casein excited at 295 nm was quenched by iodide indicating the partial exposure of tryptophan residues in protein. Addition of Fe(lll) to protein caused further quenching of tryptophanyl fluorescence by iodide. This further supports the UV spectral data that binding of Fe(lll) to protein brought about changes in the immediate vicinity of tryptophan environment. This could be due to the involvement of tryptophan residues in charge-transfer type complex formation with Fe(lll), thus exposing them to a more polar environment. Addition of Ca(ll) to a51-casein induces progressive aggregation as shown by a net increase in fluorescence emission intensity and a blue shift in fluorescence maximum. However, Fe(lll) does not seem to induce any aggregation, instead seems to form soluble Fe(lll)-protein complexes.

Fluorescence emission of a51-casein (excited at 280nm) at different pHs (5.6 to 7.8) was followed as a function of Fe(lll) concentration (0 to 0.2 mM). Addition of increasing concentrations of Fe(lll) brings about a red shift of the emission maximum, together with a decrease in fluorescence intensity at all pHs. Fluorescence intensity of a51-casein as a function of Fe(lll) concentration was fitted to a modified Stern-Volmer plot and quenching constant (Ka) was calculated from the slope. Quenching constant was high at pH 5.6 (1 .1 x1 04 M-1) and decreased with an increase in pH (0.65x1 04 M-1 at pH7.8). This indicates that Fe(lll) binding had maximum effect on the conformation of protein at pH 5.6 than at pH 7.8. Since conformational changes in general affect functional properties of proteins, it was of interest to see if binding of Fe(lll) affects Ca(ll) sensitivity of a~1-casein; addition of Fe(lll) to protein increased its Ca(ll) sensitivity .

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Impact of Research:

Iron fortification would increase the iron intakes of people who consume large amounts of dairy products, and it would allow people concerned with their nutritute to consume larger amounts of dairy products to achieve greater calcium intakes. Thus, dairy products would be even more healthful in the diet if iron-fortified. This research will provide basic information on the mechanisms of iron binding to proteins in dairy products, information essential to industrializing the technology of fortifying dairy products with iron.

Publications:

Reddy, I. M. and Mahoney, A. W. (1992). Effect of FeCI3 on chymosin hydrolysis and rennet clotting time of milk. J. Dairy Sci. (In Press).

Reddy, I. M. and Mahoney, A. W. (1992). Diafiltration and visible spectroscopic study of the binding of iron(lll) to bovine a51-. B-, and K-caseins, bovine serum albumin, alactoglobulin, and a-lactalbumin at pH 6.6. J. Agric. Food Chern. (In Review).

Abstracts:

Reddy, I. M. and Mahoney, A. W. The effect of iron on the renneting of milk. J. Dairy Sci.74 (Suppl. 1), 100, D-57, 1991.

Reddy, I. M. and Mahoney, A. W. Binding of Fe(lll) to bovine a51-casein. J. Dairy Sci.74 (Suppl. 1 ), 100, D-58, 1991 .

Reddy, I. M. and Mahoney, A. W. A study of the interaction of Fe(lll) to bovine a51-casein using ultraviolet and fluorescence spectroscopy. J. Dairy Sci. 74 (Suppl. 1 ), 100, D-59, 1991.

Reddy, I. M. and Mahoney, A. W. Characterization of Fe(lll)- milk protein complexes. In Brief Communications and Abstracts of Posters of the XXIII International Dairy Congress, Vol. I, 253, P-471, 1990.

Papers Presented at Conferences:

Reddy, I. M. and Mahoney, A. W. Effect of Iron on the renneting of milk. Presented at 86th ADSA Annual Meeting, Utah State University, Logan, August 12-15, 1991.

Reddy, I. M. and Mahoney, A. W. Binding of Fe(lll) to bovine a51-casein. Presented at 86th ADSA Annual Meeting, Utah State University, Logan, August 12-15, 1991.

Reddy, I. M. and Mahoney, A. W. A study of the interaction of Fe( Ill) with bovine a51 -casein using ultraviolet and fluorescence spectroscopy. Presented at 86th ADSA Annual Meeting, Utah State University, Logan, August 12-15, 1991.

Reddy, I. M. and Mahoney, A. W. Characterization of Fe(lll) - milk protein complexes. Presented at The 23rd International Dairy Congress', Montreal , Canada, October 7-12, 1990 .

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Project Title:

Personnel:

Funding:

Objectives:

Estimation of Individual Milk Proteins and Genetic Variants by Multicomponent Analysis of Amino Acid Profiles

Rodney J. Brown, Dept. of Nutrition and Food Sciences, Utah State University.

Carol M. Hollar, Dept. of Nutrition and Food Sciences, Utah State University.


The overall objective is to determine proportions of specific milk proteins, groups of proteins and selected genetic variants of milk and other dairy products using information obtained from amino acid analysis, fast protein liquid chromatography and isoelectric focusing of samples. Completing the following specific objectives will enable the overall objective to be met.

1. Determine concentrations of groups of proteins in milk such as caseins or whey protein and the concentrations of specific milk proteins: a51-, a52-, ~-. and K-caseins, a-lactalbumin, ~-lactoglobulin, bovine serum albumin using amino acid analysis.

2. Separate genetic variants of specific milk proteins, and use amino acid analysis to quantify individual variants in a protein mixture.

3. Use techniques developed in 1-2 to analyze milk and other dairy products.

4. Determine mathematical procedures to obtain the most accurate and reproducible methods for estimating milk protein concentrations.

Results:

In the past year, whole casein from individual cows was separated into ~-casein, x:-casein, a5 1-casein and a52-casein fractions using isoelectric focusing (IEF) and cation-exchange and anion-exchange fast protein liquid chromatography (FPLC). ~-Casein A 1, A2 and B and K-casein A and 8 all were separated using IEF. ~-Casein A1, A2 and B were separated using cation-exchange FPLC and x:-casein A and B were separated with anion-exchange FPLC. Densitometric tracings were used to quantify th,e protein peaks from the stained bands on the IEF gel. The relative proportions of the peaks were used to calculate the concentration of the caseins and selected genetic variants present in the whole casein samples. The concentrations of the caseins and selected variants were also calculated

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using the FPLC procedures. ~-Casein genetic variants were expressed equally in heterozygous samples .

Impact of Research:

The goal of this project is to determine proportions of specific milk proteins (down to the level of specific genetic variants) or groups of proteins in milk and other dairy products from the information contained in a single amino acid analysis of a sample. As profit margins get tighter, the more closely milk supplied by farmers meets the requirements of dairy manufacturers becomes more important. Manufacturers prefer receiving milk that results in improved cheese yield and processing characteristics. Developing techniques to separate and quantify caseins and their genetic variants will help provide a more desirable milk supply to manufacturers.

Publications:

Hollar, C. H., R.J. Brown and J. F. Medrano. 1990. Quantification of casein genetic variants by RP-HPLC, PAGE, and amino acid analysis. 85th American Dairy Science Association Meeting J. Dairy Sci. 73:Supp. 1, 80.

Hollar, C. M., A J. R. Law, D. G. Dalgleish, and R. J. Brown, 1991, Separation of major casein fractions using cation-exchange fast protein liquid chromatography. J. Dairy Sci. 74:2403-2409 .

Hollar, C. M., A J. R. Law, D. G. Dalgleish, and R. J. Brown, 1991, Separation of major casein fractions using cation-exchange fast protein liquid chromatography. 86th American Dairy Science Association Meeting J. Dairy Sci. 74:Supp. 1, 91.

Hollar, C. M., A. J. R. Law, D. G. Dalgleish, J. F. Medrano, and R. J. Brown, 1991, Separation and quantification of (3-casein A 1, A2 and B using cation-exchange fast protein liquid chromatography. J. Dairy Sci. 74:3308-3313.

Hollar, C. M., A J. R. Law, D. G. Dalgleish, J. F. Medrano, and R. J. Brown, 1991, Separation of (3-casein A1, A2 and Busing cation-exchange fast protein liquid chromatography. 86th American Dairy Science Association Meeting J. Dairy Sci. 74:Supp. 1, 103 .

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Project Title:

Personnel:

Funding

Objectives:

Improved Control of Cheese Manufacture Through Vat Monitoring

G. H. Richardson, Professor, Dept. of Nutrition and Food Sciences, Utah State University

Don McMahan, Professor, Dept. of Nutrition and Food Sciences, Utah State University

Michael J. LeFevre, Research Assistant, Dept. of Nutrition and Food Sciences, Utah State University.

Western Dairy Foods Research Center and Utah Agricultural Experimental Station.

1. Determine the ability of the hot-wire system to detect differences in coagulation time and curd strength. Compare these measurements with other coagulation instruments. These data would aid in preventing high moisture cheese due to late cutting or product losses due to early cutting of the curd .

2. Determine abilities of chymosin, calcium salts, and lactic cultures in milk for Cheddar cheese to overcome the inability of milk from cows in late lactation to coagulate.

3. Use the same system to monitor the coagulation of milk for cottage cheese manufacture. Establish software that would be most helpful for the cheese industry.

Results:

ADSA abstract 1992 The effects of calcium, pH and chymosin changes on the coagulation properties of late-lactation milk M.J. LeFevre*, G.H. Richardson and D.J. McMahon, Western Center for Dairy Protein Research and Technology, Department of Nutrition and Food Sciences,Utah State University, Logan.

Ten late-lactation milk samples were selected based on their coagulation properties. Upon the addition of chymosin, four samples coagulated well and six samples coagulated poorly. Fat, protein, lactose, solids-not-fat, pH and SDS-PAGE data were collected on skimmed samples. Calcium, pH and coagulant levels were adjusted using a split-split plot factorial design The pH

• of each sample was adjusted to 6.6, 6.4 or 6.2 and CaCI2 was added at .02%,

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.01% and 0% (no addition). Chymosin was added to samples at .031 and .062 rennet units/ml and a Formagraph was used to record coagulation time, K1o • K2o and relative curd firmness 30 minutes after adding chymosin. From analysis of variance we observed that all main treatments had a significant effect (p<.05) on each of the four coagulation parameters tested. Many of the interactions were also significant including chymosin x pH and calcium x pH. However, protein and lactose contents as well as protein differences shown by electrophoresis could not be correlated with milk coagulability.

Other tests have also been completed using the hot wire instrument and the Formagraph to dermine the ability of the hot wire instrument to measure coagulation time and relative curd firming rate in acid coagulated milk substrate. Software has been written that can detect relative curd firming rate, cut-time and healing time of the curd as well as temperature and pH changes during during manufacture. Relavent information has been added to software to aid in cuttime prediction. Other information relating to cut-time and relative curd strength can be saved to a file for future evalutation and addition to the software.

Impact of Research:

The data generated form such an instrument can be useful to provide improved control to every cheese vat. A curd cut time, based coagulation, pH and temperature could decrease losses and improve cheese yield and quality. The ability to measure the length of heal time and the rates of change of pH and temperature would also benefit the cheese manufacturer. Software programs could be included that would provide more management guidance. Significant savings to the industry could result when enzyme coagulant and other additive costs can be reduced by fine tuning the process through continuous monitoring of the milk in the cheese vat.

Publications Lefevre, M.J. and Richardson, G.H. 1990. Monitoring cheese

manufacture using a hot wire probe. J. Dairy Sci. 73 (suppl. 1 ):74

M.J. LeFevre, G.H. Richardson and D.J. McMahon, Use of hot-wire viscometric measurements to predict physical curd firmness of renneted milk as a function of milk composition. J. Dairy Sci. 74 (suppl. 1 ):112

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PROJECT TITLE: Cooling rate of Cheddar cheese: Comparison between 40- and 640·lb blocks and uniform cooling of 640-

Personnel:

Objectives:

lb blocks

J. Antonio Torres, Associate Professor, Dept. of Food Science and Technology, Oregon State University

Floyd W. Bodyfelt, Professor, Dept. of Food Science and Technology,

Oregon State University

Conly L. Hansen, Associate Professor, Dept. of Nutrition and Food Sciences, Utah State University

Stirling Larsen, Research Assistant, Dept. of Nutrition and Food Sciences, Utah State University

Connie Kirby, Research Assistant, Dept. of Food Science and Technology, Oregon State University

Jorge 0. Bouzas, Research Assistant, Dept. of Food Science and Technology, Oregon State University

Ricardo Simpson, Research Assistant, Dept. of Food Science and Technology, Oregon State University

Silvana Roncagliolo, Computer Research Assistant, Dept. of Computer Science, Oregon State University

Carlos A. Kantt, Research Assistant, Dept. of Food Science and Technology. Oregon State University

1. To develop a computer program that calculates cooling rate as a function of block size.

2. To use a combined hea.t transfer and microbial model to determine microbial population changes in 640-lb Cheddar cheese blocks. The microbial model had been obtained in a related project on Cheddar cheese cooling ..

3. To determine if moisture gradients observed in 640-lb Cheddar cheese ~locks are

associated with thermal gradients.

4, To determine whether a controlled cooling of Cheddar cheese blocks before cheese aging, believed to be one of the last processing steps requiring tighter control, is possible in 640-

lb blocks.

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• Results:

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Excessively high non·starter microbial counts have been reported to lead to flavor and aroma defects. Microbial models developed in a related project using linear and non-linear statistical analysis methods were used to estimate the effect of cooling conditions on non-starter counts. A combined heat transfer and microbial model was used to predict non-starter populations in various location of a 640-lb blocks undergoing cooling. Large counts were predicted in most ~ocations except those close to the block surface. Compared to non-starter counts in a 40-lb block cooled individually, the predicted values were 3 decimal order of magnitudes higher. This points to an important advantage of cooling Cheddar cheese as blocks of smaller size prior to cheese aging. A suggested area of further study would be the determination of a block size and geometry still allowing appropriate cooling rates to control non-starter microbial populations and lowering the rate of starter die-off.

An attempt to cool Cheddar cheese blocks using heat pipes demonstrated that an excessive number of such devices would have to be inserted in a 640-lb block to cool the block within a reasonable amount of time. Since both the cooling model and the use of heat pipes to lower cheese temperature-within shorter times failed, a combined computer model implemented to use calculated temperatures to estimate sensory changes and chemical indicators in a Cheddar cheese block was not used to search for cooling conditions leading to an aged cheese with desirable and homogeneous characteristics. Previous efforts to analyze the situation of palletized 40-lb blocks had demonstrated the need for early temperature control, a condition not feasible for 640-lb blocks.

Previous research efforts had demonstrated the presence of moisture gradients locked into 640-lb Cheddar cheese blocks. Experiments were set up to cool experimental Cheddar blocks from the interior (using a cooling pipe) and from the outer surface as would occur in a commercial plant. The experiment demonstrated that the moisture gradient observed during cooling from the outer surface were reversed by cooling from the inside. This demonstrates that the moisture gradients have a thermal gradient origin and further confirm the need to achieve an early and preferably uniform cooling of the cheese mass.

In summary, the project hypothesis were confirmed: early temperature control is an important parameter to reduce product variability and achieve in a consistent manner a product with a specific range of flavor quality indicators. Early temperature control seems difficult to achieve in 640-lb blocks. Future research is suggested at identifying the opdmum block geometry and dimensions desired by dairy processors for more effective handling of cheese blocks.

Impact of Research:

Cheddar cheese is. the major type of cheese produced in the United States. Sales could be further enhanced if product variability could be reduced to reduce the presence in the market of cheese

• with less desirable flavor and aroma properties.

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Publications and Abstracts

Grazier, C., McDaniel, M.R., Bodyfelt, F. W., and Torres, J .A. 1991. Temperature effects on the on the development of Cheddar cheese flavor and aroma. J. Dairy Sci.:74:3656-3669.

Bouzas, J., Bodyfelt, F.W., and Torres, J.A. 1991. A potential analytical assessment of Cheddar cheese flavor defects. Int. Dairy 1.:263-271.

Bouzas, J., Kantt, C., Bodyfelt, F.W., and Torres, J.A. 1991. Simultaneous determination of sugars and organic acids in Cheddar cheese by high-performance liquid chromatography. 1. Food Sci. 56(1):276-278.

Bouzas, J., Kantt, C.A., Bodyfelt, F.W., and Torres, J.A. Characterizar:ion and interpretation of time·temperature effects on chemical changes occurring during Cheddar cheese aging. IN REVIEW.

Grazier, C.L., Simpson, R., Roncagliolo, S., Bodyfelt, F.W., and Torres, J.A. Temperature effects on non·starter bacteria populations during cooling and aging of Cheddar cheese blocks. IN REVIEW.

Torres, J.A, Bouzas, J., Larsen, S., and Hansen, C.L. 1992. Kinetic analysis and modeling of time-temperature effects on Cheddar flavor quality. Presented at the 53rd Annual Meeting of the Institute of Food Technologists, June 20-24, New Orleans, LA.

Bouzas, J., Kantt, C.A., Bodyfelt, F.W. and Torres, J.A. 1992. Characterization of timetemperature effects on chemical changes occurring during Cheddar cheese aging. Presented at the 53rd Annual Meeting of the Institute of Food Technologists, June 20-24, New Orleans, LA.

Grazier, C.L., Bodyfelt, F.W., McDaniel, M.R., and Torres, J.A. 1991. Temperature effects on the sensory characteristics of Cheddar Cheese. Annual Meeting of the American Dairy Science Association, August 12-15, Logan, UT.

Bouzas, J., Simpson, R., Ronca.gliolo, S., and Torres, J.A. 1991. Mathematical modeling, simulation and optimization of Cheddar cheese ripening. Annual Meeting of the American Dairy Science Association, August 12-15, Logan, UT.

Bouzas, J., Bodyfelt, F.W., and Torres, J.A. 1991. Objective assessment of Cheddar cheese flavor quality. Annual Meeting of the American Dairy Science Association, August 12-15, Logan, UT.

Grazier, C.L., Simpson, R., Roncagliolo, S., and Torres, J.A. 1991. Temperature effects on non-starter bacteria populations during cooling and aging of Cheddar cheese blocks. Annual Meeting of the American Dairy Science Association, August 12-15, Logan, UT .

Bouzas, J., Kantt, C.A., Bodyfelt, F.W., and Torres, J.A. 1991. Temperature effects on

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chemical characteristics during maturation of Cheddar cheese. Annual Meeting of the American Dairy Science Association, August 12-15, Logan, UT.

Grazier, C., Simpson, R., McDaniel, M.R., Bodyfelt, F.W., Torres, J.A. 1990. Temperature effects on the microbial and sensory properties of Cheddar cheese. Annual Meeting of the Institute of Food Technologists, June 17-20, Anaheim, CA.

Bouzas, J., Bodyfelt, F.W., and Torres, J.A. 1990. Rapid and simultaneous determination of sugars and organic acids in Cheddar cheese by HPLC. Annual Meeting of the Institute of Food Technologists, June 17-20, Anaheim, CA .

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Project Title: cooling rate of Cheddar cheese: Comparison between 40 and 640 lb blocks and uniform cooling of 640 lb blocks.

Personnel: Conly L. Hansen, Professor, Dept. of Nutrition and Food Sciences, Utah State University and J. Antonio Torres, Dept. of Food Science and Technology, Oregon State University

Objectives:

1. Characterize the link between moisture movement within large Cheddar cheese blocks to the cooling rate and temperature uniformity of the block.

2. Determine the thermal conductivity of fresh Cheddar cheese curd and compare the value with the thermal conductivity of aged Cheddar cheese

3. Develop a computer model to predict internal cheese block temperature as a function of position and time (T(r,z,t), T(a,y,z,t)) when exposed to various cooling situations.

Results:

Thermal conductivity was measured with a probe which measures the conductivity of a porous material into which the probe has been placed. The Conductivity Probe consisted of a stainless steel tube in which a heating element and thermocouple were installed, imbedded in epoxy.

The heater wire and thermocouple wires were connected to extension wires and a precision resister was installed as a reference resister •

The measurement was based on the following equation:

k = q X ln(t2/tl)/(4 X pi X (T2- Tl))

where q is the power in wattsjm required to heat the probe from Tl to T2°C in the time period t2-tl sec. Therefore, if the temperature rise for a known time interval after the start of heating is determined, along with the power used to heat the probe, then the conductivity k, can be calculated. The thermal conductivity of the cheese was found to be similar to that found in the literature and used in previous modeling experiments, or about 0.35 W/m2 °C.

An experiment was devised to prove a link between temperature and moisture movement. This experiment consisted of placing a 1.3 em ID copper pipe inside a 30.5 em OD cylindrical cheese block. The results were moisture movement from outside the block to inside or just opposite the movement of moisture in large blocks of cheese.

A complex (25 page) computer model using finite difference technique was developed. Both forward and backward iterative procedures were used. The model was used to predict temperature in cheese.

Significance:

One of the major contributions of this work was to prove a link between moisture movement in cheese and temperature. Temperature has been thought to be the driving force in moisture movement which causes a moisture gradient from the inside to the outside of large blocks of cheese but it had not been demonstrated before this project was completed.

Publications:

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Personnel:

Funding:

Objectives:

Function of Whey Proteins and Lactose in Age Gelation of Ultra-high Temperature Processed Milk Concentrate.

Donald J. McMahon Venkatachalam Narayanaswamy Bashjr Yousif Douglas Olson

Nutrition & Food Sciences Dept., Utah State University

Western Dairy Foods Research Center USDA Agricultural Research Service Utah Agricultural Experiment Station

The mechanism by which age gelation in UHT processed milk concentrates occurs is still unknown. There have been many factors implicated and at best an empirical approach is taken to extend shelf life of sterilized milk products. UHT processing promotes association between K-casein and ~-lactoglobulin. In milk concentrates the concentration of whey proteins and lactose are increased. Their role in the age gelation process will be studied in this project. The objectives of the project are to:

1. Determine the influence of lactose concentration of milk concentrates on age gelation.

2. Determine the fate of (3-lactoglobulin during storage of UHT processed milk concentrates.

3. Monitor changes in casein micelle structure during storage of UHT milk concentrates.

Results:

Objective 1 : Lactose

The work on this objective has been completed. Skim was concentrated 3X by ultrafiltration and extensively diafiltered with simulated milk ultrafiltrate to remove lactose. Lactose and sucrose were then added at levels of 3 and 6% and each of the concentrates UHT processed to 140°C for 4 s and stored at 4, 20 and 35°C. Shelf stability was monitored by measuring viscosity, extent of browning was measured using a color meter and proteolysis by electrophoresis. All samples at 4 and 20°C gelled but those at 35°C did not No browning was observed in the samples containing <.1% sugars or the samples containing 3 and 6% sucrose. We concluded that sugar-protein interactions through Maillard browning reactions of lactose with proteins are not a significant factor in causing age gelation. The' streaking observed in electrophoresis of samples stored at 35°C were also not related to sugar content and therefore must relate to protein-protein interactions that occur at temperatures above 20°C. ·

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• Objective 2: 6-lactoglobulin.

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Work on this objective was postponed and planned for completion contingent upon funding of the second half of this project.

Objective 3: Casein Micelle Structure

The microstructure of UHT milk concentrates was investigated using electron microscopy in conjunction with Dr. Miloslav Kalab, Centre for Food and Animal Research, Agriculture Canada, Ottawa, Ontario, Canada and Dr. Nabil Youssef, Electron Microscopy Laboratory, Biology Department, Utah State University. It was observed that when 3X skim milk concentrate is heated to 140°C for 4 s, about 60% of the whey proteins are denatured and the casein micelles undergo a large increase in size. This size increase is due to complexing of K-casein with denatured P-lactoglobulin followed by further aggregation of denatured Plactoglobulin onto the micelle surface.

When such UHT samples are stored and eventually age gel, it was observed that many of the micelles were connected by thin threads of material. The microstructure of such gels was completely different to rennet milk gels. When rennet is added to non-UHT milk the casein micelles collide and form a gel structure in which the micelles are in close contact. However, if UHT milk concentrates (such as 3X UF concentrates) are renneted the gels have less integrity and there is not the same level of intimate contact between micelles. It appears that the layers of P-lactoglobulin on the micelle surface interfere with aggregation.

Impact of Research:

The overwhelming success of dairy production in the United States has created a situation that calls for more attention to milk utilization and marketing. Development of new dairy products has now taken precedence over production of more milk.

International markets for U.S. dairy products could be developed if attention was directed to manufacture of stable products from our surplus dairy production. The competitive position of the U.S. would be enhanced by new and better quality products and our surpluses of dairy commodities would be reduced.

The major limitation to production of such a product is irreversible gelation after storage at higher-than-refrigerator temperatures over a long period of time. The gelling phenomena must be understood and means devised to prevent gel formation before rehydratable milk concentrates can be sold abroad.

Publications: Venkatachalam, N., D.J. McMahon, and P.A. Savello. The role of lactose in age

gelation of ultra-high temperature processed UF milk concentrates. J. Dairy Sci. (Submitted).

McMahon, D.J., B. H. Yousif and M. Kalab. 1992. Effect of whey protein denaturation on structure of casein micelles and their rennetability after ultrahigh temperature processing of milk . Int. Dairy J. (In press).

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Venkatachalam N., 1992. The role of lactose in the age gelation of ultra-high temperature processed concentrated milk, M.S. Thesis, Utah State University.

Yousif, B. 1991. Effects of heat treatment of ultrafiltered milk on its rennet coagulation time and whey protein denaturation, M.S. Thesis, Utah State University.

Olson, D.W. and D.J. McMahon. 1992. Zeta-potential of casein micelles as a factor in age gelation of ultra-high temperature sterilized concentrated milk. 87th American Dairy Science Association Meeting, J. Dairy Sci. 75{Supp. 1 }:

Venkatachalam, N. and D.J. McMahon.. 1992. The effect of microbial quality on the storage stability of ultra-high temperature processed skim milk concentrate. 87th American Dairy Science Association Meeting, J. Dairy Sci. 75(Supp. 1 }:

Venkatachalam, N. and D.J. McMahon.. 1992. Effect of ultrafiltration, diafiltration and ultra-high temperature processing on casein micelle size. 87th American Dairy Science Association Meeting, J. Dairy Sci. 75(Supp. 1}:

Olson, D.W. and D.J. McMahon. 1991. Zeta-potential of casein micelles in UHTtreated ultrafiltered skim milk. 86th American Dairy Science Association Meeting, . Dairy Sci. 74(Supp. 1 }:D155

Kalpalathika, P.V.M., N. Venkatachalam and D.J. McMahon. 1991. The effect of storage of UHT sterilized skim milk concentrates on their zeta-potential, particle size, browning and viscosity. 86th American Dairy Science Association Meeting, J. Dairy Sci. 74(Supp. 1 }:D156.

Venkatachalam, N. and D.J. McMahon. 1991. Effect of lactose concentration on age gelation of UHT sterilized skim milk concentrate. 86th American Dairy Science Association Meeting, J. Dairy Sci. 74(Supp. 1 }:D62 .

Yousif, B.H., D.J. McMahon, P.A. Savello, R.J. Brown and M. Kalab. 1990. Effects of heat treatment of concentrated milk on its rennet coagulation time, gel microstructure and whey protein denaturation. 85th American Dairy Science Association Meeting, J. Dairy Sci. 73 (Supp. 1 }:1 05 .

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Project Title:

Personnel:

Funding:

Objectives:

Controlling Age Gelation of UHT processed milk concentrates.

Donald J. McMahon Mohamed Mohamed Mrudula Kalpalathjka Nutrition & Food Sciences Dept., Utah State University

Western Dairy Foods Research Center USDA Agricultural Research Service Utah Agricultural Experiment Station

Age gelation of ultra-high temperature (UHT} milk concentrates has hindered the commercial use of milk concentration as a means of lowering transport costs. These has been some work conducted on developing stable UHT milk concentrates but at best, an empirical approach is taken to extend shelf life. Composition of milk, severity of heat treatment, sequence of operation, homogenization, use of additives such as polyphosphates and sucrose, total solids, and enzyme treatment have all been shown to affect shelf life of UHT milk C0!1centrates. The specific objectives of this project are to:

1. Determine influence of compositional changes of milk concentrates on age gelation after UHT processing .

2. Determine effects of process parameters of UHT heating on age gelation of milk concentrates.

3. Use the data collected above to determine optimum conditions for UHT processing of milk concentrates.

Results: Casein micelle size of UF concentrated milk was measured before and after UHT processing using photon correlation spectroscopy. Ultrafiltration and diafiltration (with Simulated Milk Ultrafiltrate) did not significantly affect particle size whereas UHT processing did increase the size of casein micelles. The size increased from 175 nm before UHT treatment to 355 nm in the undiafiltered sample and to 258 nm in the diafiltered samples. Upon prolonged storage after UHT processing, the undiafiltered sample formed a sediment at the bottom of the container while the diafiltered samples formed either a soft gel at the bottom of the container or a gel uniformly distributed throughout the container. This difference in the storage behavior was related to the difference in. size of the casein micelles immediately after processing.

Skim milk was concentrated by UF to 2X and 3X by reverse osmosis (RO) to 2X. Lactose was added to a part of the 2X UF concentrate (50 g/L) to provide a lactose concentration comparable to the 2X RO sample All the samples were UHT processed at 140°C for 4 s. Micellar zeta-potential and size, pH, viscosity, calcium

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• and phosphorus and SDS-PAGE data were collected on samples stored at 35°C, 20°C, and 4°C at an interval of 45 days for one year of storage. Gradual decrease in magnitude of micellar zeta-potential (-18 mV to -14 mV) and increase in size {320 nm to 520 nm) were observed in all samples stored at all temperatures, the changes being larger in samples concentrated to 3X by UF and 2X by RO. SDSPAGE electrophoresis patterns indicated occurrence of proteolysis in all samples irrespective of storage temperatures, with streaking observed, at 35°C. Soluble calcium decreased by 1 to 2 mM in 1 X and 2X UF samples, 4 mM in 3X UF samples and 5 mM. in 2X RO samples. However, the decease in soluble phosphorus content was minimal in all samples. 3X UF and 2X RO gelled after 9 months of storage at 35°C and 20°C. 2X UF samples remained stable at 20°C and 4°C for 12 months. Excessive browning occurred in samples stored at 35°C.

Impact of Research: The overwhelming success of dairy production in the United States has created a situation that calls for more attention to milk utilization and marketing. Development of new dairy products has now taken precedence over production of more milk. At the same time, we are becoming increasingly aware that a global approach must be taken in marketing our dairy products. Two marketing strategies that are of importance are Export and Military. In both cases, transportation costs are very high. By reducing the bulk of a product, such as by concentrating milk, transportation costs can be reduced and furthermore, for many military applications it would alleviate space storage limitations of supplying "fresh" milk to military personnel.

• International markets for U.S. dairy products could be developed if attention was directed to manufacture of stable products from our surplus dairy production. The competitive position of the U.S. would be enhanced by new and better quality products and our surpluses of dairy commodities would be reduced.

Specifically for this project, a way of producing rehydratable milk concentrates that can be stored at ambient temperatures is to be developed. This is needed to make U.S. dairy products more widely available on the global market. The major limitation to production of such a product is irreversible gelation after storage at higher-than-refrigerator temperatures over a long period of time. The gelling phenomena must be understood and means devised to prevent gel formation before rehydratable milk concentrates can be sold abroad.

Publications: McMahon, D.J., B.H.Yousif and M. Kalab. 1992. Effect of whey protein

denaturation on structure of casein micelles and their rennetability after ultrahigh temperature processing of milk . Int. Dairy J. {In press).

McMahon, D.J., P.A. Savello, R.J. Brown and M. Kalab. 1991. Effects of phosphate and citrate on gelation properties of casein micelles in renneted ultra-high temperature (UHT) sterilized concentrated milk. Food Structure 10 {1 ):27.-36.

Kalpalathika, P.V.M. and D.J. McMahon. 1992. Study of molecular zeta-potential and size and calcium and phosphorus contents of UHT processed skim milk concentrates during storage. 87th American Dairy Science Association

• Meeting, J. Dairy Sci. 75{Supp. 1):

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Project Title: Using Whey For Improvement of Exposed Subsoils and Sodic and Saline-Sodic Soils

Personnel: Conly L. Hansen, Professor, Dept. of Nutrition and Food Sciences, Utah State University

Objectives:

Main Objective

Determine if land application of whey is beneficial to soils, particularly sodic and saline-sodic soils.

Specific Objectives

-Determine the extent crops and soil removes nutrients and lactose from whey.

-Determine maximum safe whey land application rate.

-Determine if odor will be a problem in land application of whey and/or how to avoid an odor problem

-Determine value of whey for land treatment, ie from nutrients, pH adjustment, and soil tilth

Results:

Reclamation abstract

Cottage cheese production in the u.s. yielded approximately three million Mg of cottage cheese (acid) whey in 1991. Unmarketable whey is disposed of in sewage treatment facilities or on land. Environmental concerns and new laws make disposal even more costly and difficult. While much whey is applied to land for fertilizer or disposal purposes, acid whey has recently been used as a sodic soil amendment. The objectives of this research were to determine the effects of whey application on chemical properties and infiltration rates of sodic soils. Four (0, 2S, SO, and 100 mm) treatments of acid whey were applied to Freedom silt loam (fine-silty, mixed, mesic, Xerollic Calciorthids) in greenhouse lysimeters and to Declo loam (coarse-loamy, mixed, mesic, Xerollic Calciorthids) in field basins. Accumulative sodium removal at O.S pore volumes of leachate was 1.01, 1.14, 1.23, and 1.72 moles for the O, 2S, SO, and 100 mm lysimeter treatments, respectively. Whey applications lowered sodium adsorption ratio (SAR), exchangeable sodium percentage (ESP), and saturation paste pH in the lysimeter surface soil (0-1SO mm) and field soil surface (10-1SO mm). Saturated paste electrical conductivity (EC) was not significantly altered by whey application. In a greenhouse environment, 28 days after whey application, infiltration times in the 2S, SO, and 100 mm treatments were 324, 434, and 416 percent of the 0 mm treatment, respectively. Infiltration times for field basin 2S, SO, and 100 mm whey treatments were, 33, 31, and 26 percent of the 0 mm treatment 53 days after whey application, respectively. Barley (Hordeum vulgare L. 'Ludd') grain yield was 0.0, 0.0, 0.44, and 0.26 kg m·2 and total dry matter yield was 0.54, 0.72, 2.04, and 1.43 kg m·2 for the 0, 2S, SO, and 100 mm treatments, respectively. Excess salts and/or organic overloading in the 100 mm treatment appeared to inhibit initial plant growth. Results indicate acid whey is effective in reclaiming sodic soil by lowering ESP, SAR, and pH and by improving infiltration.

COD abstract

Cottage cheese reclamation, however,

(acid) whey is an effective amendment in sodic ·soil the high chemical oxygen demand (COD) of whey is of

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concern in land application. The objective of this research was to determine the fate of COD from cottage cheese whey applied to a sodic soil. Treatments of 0, 25, 50, and 100 mm (0, 20, 40 and 80 Mg COD ha"1

) of whey were applied to dry-unacclimated Freedom silt loam (fine-silty, mixed, mesic, Xerollic Calciorthids) in greenhouse lysimeters. The COD from lysimeter leachate at 1 m depth was monitored. Ninety days after whey application, total accumulative leachate COD for 0, 25, and 50 mm whey applications was not significantly different. Leachate COD concentrations from the 100 mm application reached 37 percent of the applied whey COD. Twenty eight days after whey treatment, infiltration was reduced in all whey-treated lysimeters, probably as a result of increased microbial activity. Barley (Hordeum vulgare L. 'Ludd') grain yield was 0.0, 0.0, 0.44, and 0.26 kg m·2 and total dry matter yield was 0.54, 0. 72, 2.04, and 1.43 kg m·2 for the O, 25, 50, and 100 mm treatments, respectively. Salts and/or organic overloading appeared to inhibit initial barley growth in the 100 mm treatment. Results indicate a single 100 mm application to be excessive in terms of organic matter and/or salts.

Significance:

Cottage cheese (acid) whey was shown to be a beneficial sodic soil amendment. Whey acidity, soluble ca, Mg, and K, and organic matter should all contribute to soil reclamation. Increasing the depth of whey application lowers the soil SAR, ESP, and saturation paste pH. Results indicate sodium removal is proportional to the amount of whey applied. The concentrated chemical oxygen demand of cottage cheese (acid) whey must be considered in land application. Increased microbial activity from whey organic matter may initially decrease infiltration rates. Given adequate resting periods, whey application followed by leaching appeared to improve infiltration. Crop yield is improved by whey application if COD or salt overloading does not inhibit growth. Depth of application should be limited by organic loading of soil and possible groundwater contamination from nitrates and other nutrients. The heavy chemical oxygen demand of a 100 mm application was excessive in greenhouse lysimeters while applications of 25 and 50 mm were not significantly different from the 0 mm whey treatment.

Publications:

1. Jones, S.B., c. w. Robbins, C.L. Hansen. Reclamation Using Cottage Cheese (Acid) Whey. and Rehabilitation. Submitted May 1992.

1992 Sodic Soil Arid Soil Research

2. Jones, S.B., C.L. Hansen, c. w. Robbins. 1992 Fate of Chemical Oxygen Demand from Cottage Cheese (Acid) Whey Applied to a Sodic Soil. Arid Soil Research and Rehabilitation. Submitted May 1992 .

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• Project Title:

Personnel:

Cloning the Nisin and Other Genes of Lactic . Streptococci into Leuconostoc Species and Amplification of Nisin Production

W. E. San dine, Principal Investigator, Dept. of Microbiology, Oregon State University

M. Levata-Jovanovic, Graduate Research Assistant, Dept of Microbiology, Oregon State University

H. A. Wyckoff, Graduate Research Assistant, Dept. of Microbiology, Oregon State University

Funding Sources: Western Dairy Foods Research Center

Objectives: 1. To produce and characterize lactose positive Leuconostoc transconjugants

obtained by conjugal matings between Lactococcus lactis and Leuconostoc spp.

2. To develop transformation and gene cloning systems in Leuconostoc.

3. To· introduce into Leuconostoc, plasmid-coded-protease genes from lactic streptococci.

• 4. To use the genetically constructed fast acid-producing Leuconostoc to produce different fermented dairy products such as cultured buttermilk,

cottage and Gouda cheese.

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5. To conjugally transfer plasmid-coded nisin genes from Lactococcus lactis to Leuconostoc.

6. To amplify nisin production by gene cloning techniques.

7. To. use nisin producing Leuconostoc in Swiss cheese manufacture to inhibit gas producing anaerobic spore formers such as Clostridium tyrobutyricum.

8. To study the inhibition of L. mpnocytogenes by nisin and. to use genetic engineering to maximize its useful application.

Results : Briefly reviewing progress reported in previous years; The results of a study demonstrating the efficacy of nisin inhibition of Listeria ' monocytogenes have been published (Benkerroum and Sandine;1988). We have published an article describing a genetic transformation system which is applicable to all species of dairy Leuconostoc and identified a

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series of cloning and shuttle vectors which are functional in these organisms (Wyckoff, Sandine, and Kondo ;1991) .

Previously we reported on the construction of a stable cloning vector for use in Leuconostoc. Presently, we are completing the final stage of construction which will produce a 3.5 kb plasmid containing 18 unique restriction sites and the resistance gene to the antibiotic chloramphenicol. We have used the published DNA sequences of the Streptococcus thermophilus lactose permease (lacS) and B-ga1actosidase (lacZ) to construct oligonucleotide primers and amplify the two genes on a contiguous DNA fragment using the polymerase chain reaction. Using unique restriction sites within the primers, we were able to clone the fragment into an E. coli cloning vector and subsequently subcloned it into vectors which are functional in lactic acid bacteria. Leuconostoc strains with these clones express high levels of Bgalactosidase but are not able to transport lactose into the cells. Efforts are continuing to achieve expression of both genes in Leuconostoc.

Impact of Research: The success of transforming and expressing the heterologous Streptococcus

thermophilus B-galactosidase in Leuconostoc shows that strains of these organisms can be developed which integrate characteristics of other starter bacteria. Such strains would be expected to carry out more reliable dairy fermentations or could be used to produce new specialty dairy products.

Publications:

Mahmoud, H. M. A., W. E. Sandine and J. K. Kondo. 1988. Lactose use by genetically manipulated Leuconostoc. J. Dairy Science. 71:Suppl 1 Abst D-4

Benkerroum, N. and W. E. Sandine. 1988. Inhibitory action of nisin against Listeria monocytogenes. J. Dairy Science. 71:3237-3245

Wyckoff, H. A., W.E. Sandine, and J. K. Kondo. 1989. Electro-transformation of Leuconostoc with plamid DNA. J. Dairy Science. 72:Suppl 1 Abst

Kondo, J. K., H. A. Wyckoff, and W. E. Sandine. 1989. Manipulation of Leuconostoc for improved milk fermentation. Soc. Ind. Microbiol. Annual Meeting Abst.

Wyckoff, H. A., W. E. Sandine, and J. K. Kondo. 1991. Transformation of D~iry Leuconostoc using plasmid DNA from Bacillus, Escherichia, and Lactococcus hosts. J. Dairy Science. 74:1454-1460 .

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• Project Title: Cloning the Nisin and other Genes from Lactococcus

into Leuconostoc Species and Amplification of Nisin

Personnel:

Funding:

Production.

William E. Sandine, Professor, Department of Microbiology, Oregon state University, Corvallis.

Jeffery R. Broadbent, Assistant Professor, Department of Nutrition and Food Sciences, Utah State University, Logan.

Herb Wycoff, Graduate Research Assistant, Dept. Microbiology, osu.

Hua Wang, Graduate Research Assistant, Dept. NFS, usu.

Yat-Chen Chou, Graduate research Assistant, Dept. NFS, USU.

Western Dairy Foods Research Center Utah Agricultural Experiment Station

Objectives: 1. To produce and characterize lactose positive Leuconostoc

transconjugants obtained by conjugal matings between • Lactococcus and Leuconostoc spp.

2. To develop transformation and gene cloning systems in Leuconostoc.

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3. To introduce into Leuconostoc, plasmid-coded protease genes from lactococci (e.g. L· lactis subsp. cremoris Wg2).

4. To use the genetically constructed fast acid-producing Leuconostoc (lactose positive and protease positive) to produce different fermented dairy products such as cultured buttermilk, cottage and Gouda cheese.

5. To conjugally transfer plasmid-coded nisin genes from L· lactis subsp. lactis 7962 to Leuconostoc and to study this plasmid at the molecular level.

6. To amplify nisin production by gene cloning techniques.

7. To use nisin producing Leuconostoc in Swiss cheese manufacture to inhibit gas producing anaerobic sporeformers such as Clostridium tyrobutyricum.

8. To study the inhibition of L· monocytogenes by nisin and to use genetic engineering techniques to maximize its useful application.

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Results, utah State University:

During the past year, our research has focused on objectives 5 and 6. We have studied nisin-sucrose conjugation in a lactococcal model system to develop methodology which will permit transfer into Leuconostoc spp. and other lactic acid bacteria. From these experiments we have developed an improved conjugation technique, DPC, which has allowed us to transfer nisin-sucrose genes into strains of ~. lactis subsp. cremoris and Streptococcus salivarius subsp. thermophilus. The streptococcal transconjugants expressed nisin immunity and sucrose fermenting ability (Sue+) but, unlike lactococcal transconjugants, they did not produce nisin (Nip+). DNA-DNA hybridizations, however, demonstrated that the n1.s1.n structural gene was present in streptococcal transconjugants. As we indicated last year, intergeneric transfer appeared to rely on the presence of the broad host-range conjugative plasmid p~1 (5) in streptococcal recipients. The ability of this plasmid to facilitate conjugation in trans was further demonstrated by Sue+ transfer from lactococcal donor DL16 (3) exclusively to recipients that contained pAMP1. Unfortunately, we were not able to detect n1.s1.n-sucrose transfer to Leuconostoc or Lactobacillus spp. recipients that contained pAMP1. Because of this difficulty, we have started to investigate protoplast fusion as a method to transfer nisin-sucrose genes into dairy lactic acid bacteria. We have been able to remove and regenerate the cell wall from lactococci and Leuconostoc mesenteroides subsp. dextranicum 181 (4) and fusion experiments are presently underway.

During the past year we completed our investigation into the location of the nisin genes in Lactococcus lactis subsp. lactis 7962, DL16, and 11454. As reflected in objective 5, we initially believed that Nip+Suc+ was encoded by a 14.5 MDa plasmid in strain 7962 (4). Several investigators had also associated Nip+Suc+ to a 29.1 MDa plasmid in strain 11454 ( 1, 2, 3) . Plasmid curing, conjugations, and DNA-DNA hybridizations demonstrated that the nisin structural gene was not associated with detectable plasmid DNA in any of our Nip+ strains.

As reported last year, we have cloned a 10 kb fragment that contained the nisin gene and introduced the recombinant plasmid, pJB100, into Lactococcus lactis subsp. lactis LM0230. Phenotypic analysis of electrotransformants did not reveal expression of nisin production or immunity, sucrose utilization, or reduced phage sensitivity. We conjugally introduced Nip+Suc+ into a strain that contained pJB100 to determine whether we might obtain a gene dosage effect, but we were not able to detect a change in the level of nisin production. We are continuing our efforts to clone, genes which will allow us to express nisin production or immunity in lactic acid bacteria .

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Impact of Research - Nisin:

Nisin, a peptide antibiotic produced by some strains of Lactococcus lactis subsp. lactis, is an effective inhibitor of Gram-positive bacteria. The antibiotic has been approved for use as a food preservative in over forty-five countries, including the United States. For years, investigators have been interested in transferring the nisin genes into other organisms used to manufacture fermented foods to enhance the shelf life of these products. Conjugation, a natural gene transfer process, may be useful to achieve this goal. Bacterial strains that are developed via conjugation contain genes that only come from other safe, food-grade lactic acid bacteria. Consequently, conjugally improved strains may face fewer FDA restrictions, with respect to industrial application, than strains which are improved through recombinant DNA technology.

Our studies of conjugation have yielded new methodology which has allowed us to investigate many of the parameters which affect transfer of the nisin genes. This work has led to improved transfer efficiency and the subsequent ability to transfer these genes into strains of Lactococcus lactis subsp. cremoris and Streptococcus salivarius subsp. thermophilus. These results indicate that we may be able to use conjugation to introduce the nisin genes into leuconostoc and other genera of lactic acid bacteria. We can foresee many applications for nisin-producing lactic organisms among dairy, food, and agricultural fermentations, to enhance the safety and shelflife of foods.

construction of rapid acid-producing leuconostoc. Leuconostoc spp. are slow acid-producers in milk and thus are

unable to produce appreciable amounts of the important flavor compound, diacetyl, in pure milk cultures. This shortcoming has been attributed to their limited ability to metabolize milk proteins and lactose. If the growth of Leuconostoc spp. is slow or inhibited, the fermented product lacks proper flavor. By increasing the levels of proteolysis and lactose utilization, through gene transfer from lactococci to Leuconostoc spp., we may be able to reduce or eliminate flavor defects in milks fermented with Leuconostoc spp. Genetically modified Leuconostoc spp. may also have application for the manufacture of specialty cheeses similar to varieties now imported, or the development of novel dairy products .

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Publications: Broadbent, J.R. and J.K. Kondo. 1990. Genetic construction of nisin-producing L· lactis ssp. cremoris and analysis of a rapid method for conjugation. Appl. Environ. Microbial. 57:517-524.

Broadbent, J.R., J.K. Kondo and W.E. sandine. 1992. Location of the nisin genes in the producer organism. Submitted to J. Food Protect.

Broadbent, J.R., J.K. Kondo and W.E. Sandine. 1992. Characteristics of nisin-sucrose conjugal transfer and intergeneric transfer of genes associated with nisin production. Submitted to Appl. Environ. Microbial.

Abstracts: Broadbent, J.R. and J.K. Kondo. 1990. Genetic construction of nisin-producing strains of Lactococcus lactis ssp. cremoris. J. Dairy Sci. 73 (Suppl. 1):72.

Wang, H., and J.K. Kondo. 1991. Study functional characteristics of cell transconjugants of Lactococcus lactis Dairy Sci. 74 (Suppl. 1):25.

References:

of the physical and clumping in Lac+

ssp. lactis ML3 . J.

1. LeBlanc, D.J., V.L. Crow, and L.N. Lee. 1980. Plasmid mediated carbohydrate catabolic enzymes among strains of Streptococcus lactis p. 31-41. In c. Stuttard and K.R. Rozee ( ed. ) , Plasmids and transposons: environmental effects and maintenance mechanisms. Academic Press Inc., New York, NY.

2. Gonzales, C.F., and B.S. Kunka. 1985. Transfer of sucrose-fermenting ability and nisin production phenotype among lactic streptococci. Appl. Environ. Microbial. 49:627-633.

3. Steele, J.L., and L.L. McKay. 1986. Partial characterization of the genetic basis for sucrose metabolism and nisin production in Streptococcus lactis. Appl. Environ. Microbial. 51:57-64.

4. Tsai, H.-J., and W.E. Sandine. 1987. Conjugal transfer of nisin plasmid genes from Streptococcus lactis 7962 to Leuconostoc dextranicum 181. Appl. Environ. Microbial. 53:352-357.

5. Vescovo, M., L. Morelli, v. Bottazzi, and M.J. Gasson. 1983. Conjugal transfer of broad-host-range plasmid pAM/31 into enteric species of lactic acid bacteria. Appl. Environ. Microbial. 46:753-755 .

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Project Title:

Personnel:

Funding Sources:

Objectives:

Final Report

Purification of a bacteriocin from Pediococcus pentosaceus arid genetic transfer of the plasmid borne determinants 7/1/88-12/31/91

Mark A. Daeschel, Principal Investigator, Dept. of Food Science and Technology, Oregon State University

Xintian Min~, Graduate student, Dept of Food Science and Technology, Oregon State University


The primary objectives of this study were twofold:

1. To purify the bacteriocin, Pediocin A, using protein purification methodology to a purity suitable for the production of polyclonal antibodies.

2. To genetically transfer the Pediocin A plasmid (p:MD136) into dairy fermentation strains via the current state of the genetic transfer systems that have been demonstrated with lactic acid

·bacteria

Potential Impact of the Research:

Modern genetics has provided the tools for constructing lactic acid bacteria with the ability

to enhance the quality of fermented dairy foods and to provide a higher degree of preservation.

Genetic transfer systems such as transformation, transduction and conjugation have been

demonstrated in lactic acid bacteria and are currently being used by researchers in dairy starter

cultures for strain improvement. These technologies will allow us to transfer into dairy cultures the

genes that encode for antagonistic substances (bacteriocins) that are active against a variety of

undesirable spoilage and pathogenic microorganisms that occur in fermented dairy foods. The

inhibition of undesirable microorganisms will allow for enhanced product quality and safety which

in the long run will enhance milk utilization.

Of current concern to cheese processors are the occurrence of microorganisms which cause

blowing faults (Clostridia tyrobutyricum) in Swiss-type cheese and cheeses. Bacteriocin producing

starter cultures is one _approach for controlling the incidence of such microorganisms in fermented

dairy foods. Certain bacteriocins from non-dairy lactic acid bacteria have been shown to inhibit

clostridia and Listeria as well as other pathogens such as Staphylococcus aureus.

The acquisition of bacteriocin producing ability by dairy starter cultures through genetic

47

biotechnology may allow the development of strains superior to those presently available.

• Results:

•

•

Initial studies were focused on concentration and purification of Pediocin A to the extent where it

could be incorporated into a selective medium suitable for the recovery of transformants. We have

not been able to demonstrate activity in culture supernatants in which Pediocin A producing

· Pediococcus pentosaceus has been cultivated. Ultrafiltration or dialysis concentration of

supernatants did not result in detection of Pediocin A activity when assayed by the well diffusion

method. However, activity is clearly seen with the deferred antagonism method using the same

medium (Trypticase Soy Broth). We also attempted to develop a selection system for recovering

potential Pediocin A transformants by utilizing the antagonistic may have on untransformed

recipients. Previous data (Daeschel, unpublished) has shown the frequency of (bac-) cells

harboring pMD 136 to range from .001% to 99.9% depending on temperature of cultivation.

Populations that were predominantly (bac-) after cultivation at 42 C became predominantly (bac+)

after several growth transfers at 37C. This may reflect the "self-selecting" of the bacteriocin

immu~ity genotype where plasmid (pMD136) cured cells may be vulnerable to the bacteriocin. We

attempted to utilize these observations to develop a selection system for Pediocin A transformants

based on their ability to predominate over untransformed cells after several growth cycles. We

were uP-successful in using this approach to select (bac+) transformants and decided to pursue a

different approach.

During the 2nd year we concentrated on optimizing electroporation transformation

procedures for Pediococci and Lactococci with the view of introducing the Pediocin A plasmid

since we are unable at that time to use Pediocin A as a selective agent. Two alternative approaches

are being explored. The first approach is to optimize electroporation conditions so that co

transformation ofpMD136 and a directly selectable marker (antibiotic resistant plasmid) could

occur. Co-transforrnants could then be individually screened for Pediocin A production. The

second approach is to clone into a directly selectable plasmid vector the Pediocin A genes for

production and immunity to Pediocin A. Both approaches are dependent on high transformation

frequencies. We have been able to transform 3 of 5 strains ofpediococci using the plasmid vector

pNZ12 (table 1). Transformation frequencies were in the range of 2-3 X 102 per ug of DNA. We

have also been able to obtain transformation frequencies of about 1 X 106 per ug of DNA with

Lactococcus lactis LM 2302 which will be used as a recipient for pMD 136- pNZ12 co

transformation experiments. The data from the L. lactis experiments may be useful for other

investigators and as such is presented in figures 1-4. Co-transformation experiments using pMD

136 and pNZ12 did not yield any selectable transforrnants carrying pMD136 .

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During the last year of the project we have redoubled our efforts toward isolating active

Pediocin A. Various media formulations were evaluated in both deferred antagonism and

supernatant assays. We still have been unable to make significant progress in isolating enough

active Pediocin A in sufficient quantities for use in plasmid transformation experiments. It appears

that Pediocin A may be a very fragile protein in terms of maintaining bioactivity. We believe that

physical manipulations may accelerate loss of activity.

The desired objectives of the project were not achieved to our satisfaction. Fundamental

problems impeded a successful conclusion to the project The inability to purify pediocin A and the

lack of a suitable transformant selection system were the two major problems. We believe we had

exhausted to our capability and expertise any feasible experimental avenues. We do not feel that a

continuation or renewal of this project by us or others would produce results directly applicable to

the dairy industry in a reasonable time period.

Table 1 . Transformation of Pediococcus by

electroporation

Strains Transformation frequency (CFU/ug DNA)

P. cerevisiae PC-39 0

P. pentosaceus B-11456

P. pentosaceus 257 45

P. pentosaceus 33314

P. pentosaceus FBB-61-2

49

200

0

200

300

••

I 0 I I I 4

g. 1 The effect of field. strength on transformation frequency

of L.lactis LM0230.

0'1 0

6.0

5.8

< z 0 5.6

-a- log CFU/ugDNA Ol ~ :::> u. 5.4 u Ol .2

5.2

5.0 0.2 . 0.4 0.6 0.8 1.0

o.o soo

•

Fig. 2

ig. 3 The effect of growth phase on transformation· frequency Fig. 4

of L.lactis LM0230.

< z 0 Ol

~ u. u Ol

.3

• 6

5 -a- LogCFU/ugDNA

4+-~~--~~~~~~~~~

0 200 400 600 800 1000

R (ohm)

The effect of resistence on transfom1ation frequency of

L.lactis LM0230.

-a- Log CFUtugDNA

The effect of glycine on transformation frequency of L.Lactis LM0230.

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Project Title:

Personnel:

Funding:

Objectives:

Generation of Molecular Probes for Bifidobacterial Species

Joseph W. Booth, Dept. of Biochemistry and Biophysics, Oregon State University

Janine E. Trempy, Dept. of Microbiology, Oregon State University

William E. Sandine, Dept. of Microbiology, Oregon State University


There is a great potential market in this country for the sale of fermented dairy products containing healthful bacterial cultures. The research we have proposed will enable accurate quantitation of viable cell counts of bifidobacteria in dairy products containing mixed cultures. We have proposed a molecular approach to the detection of bifidobacterial cells:

1. Development of an antibody against fructose-6-phosphate phosphoketolase. Fructose-6-phosphate phosphoketolase is an enzyme unique to bifidobacteria. Antibody production necessitates purification of the enzyme in quantities sufficient for injection into rabbits. With the antibody in hand, Western blots of colony lifts of plated cultures from fermented dairy products can be performed.

2. Development of a genomic probe against the gene for fructose-6-phosphate phosphoketolase. Achievement of a homologous genomic probe necessitates the cloning of the gene for fructose-6-phosphate phosphoketolase. E. coli cells containing the cloned gene can be identified from a library of cloned sequences by screening the library with the antifructose-6-phosphate phosphoketolase antibody.

Results:

The enzyme fructose-6-phosphate phosphoketolase has been purified to homogeneity as visualized by SDS polyacrylamide gel electrophoresis.

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• There are three steps in the purification scheme. After an initial precipitation with protamine sulfate to remove nucleic acid and unwanted crude protein, 20 ml of a 5 mg/ml extract are applied to a 2.6 x 100 em Sephacryl S-300 column. Fructose-6-phosphate phosphoketolasecontaining fractions are pooled and next applied to a Mono-0 FPLC column and eluted with a 100 mM NaCI to 500 mM NaCI gradient. Fructose-6-phosphate phosphoketolase elutes as two peaks from the Mono-Q column. The first peak contains the purified enzyme; the second peak contains enzyme contaminated with a cellular protein slightly smaller in size than fructose-6-p hosphate phosphoketolase.

Antisera against fructose-6-phosphate phosphoketolase has been generated by injection of a rabbit with the purified enzyme.

The N-terminal 30 amino acids of fructose-6-phosphate phosphoketolase have been determined by solid phase amino acid sequencing of the purified enzyme. A stretch of 5 amino acids with minimum degeneracy (48-fold) coding for a 15 nucleotide oligomer have been targeted as a potential probe for the fructose-6-phosphate phosphoketolase gene.

Impact of Research:

• Generation of molecular probes for the identification of bifidobacterial species will make possible the accurate determination of numbers of viable bifidobacterial cells in mixed cultures containing bifidobacteria and other lactic acid bacteria. This will facilitate marketing a variety of bifidobacterial-containing milk products of uniform quality where numbers of these bacteria are concerned.

Publications:

None

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Project Title:

Personnel:

Funding:

Objectives:

The Influence of Preadsorbed Protein on Adhesion of Listeria monocytogenes to Dairy Food Contact Surfaces

M.A. Daeschel, Associate Professor, Department of Food Science and Technology, Oregon State University

J. McGuire, Assistant Professor, Departments of Bioresource Engineering and Food Science & Technology, Oregon State University

H. AI-Makhlafi and C.K. Bower, Research Assistants, Department of Food Science and Technology, Oregon State University

Western Dairy Foods Research Center, US Agency for International Development, Public Health Service Institutional Grant, and Oregon Agricultural Experiment Station

· 1. record ~-lactoglobulin adsorption kinetic data on each of a series of silanized silicon surfaces that have been prepared to exhibit varying degrees of hydrophobic and hydrophilic character;

2. compare the measured adsorption kinetics to that predicted by a simple . molecular model of interfacial behavior to obtain a measure of the rate at which conformational changes take place on each surface;

3. prepare adsorbed ~-lactoglobulin layers of varying age on each type of surface, and quantify the adsorbed mass of each film;

4. expose the surfaces, with and without preadsorbed films, to Listeria monocytogenes in order to document the relationship between characteristics of each protein film and the extent and tenacity of resulting microbial adhesion; and

5. model the extent and tenacity of microbial adhesion as a function of contact surface hydrophobicity, or the nature of the preadsorbed film.

Results:

Project objectives 1 through 3 were· completed. The adsorption kinetics exhibited by a-lactalbumin (a-lac), ~-casein, and bovine serum albumin (BSA) at hydrophilic and hydrophobic silicon surfaces were recorded with ellipsometry as well, and interpreted with reference to a simple mechanism for irreversible protein adsorption. These additional tests were performed in response to TAC recommendations, and constitute a natural extension of past work that involved measurement of ~-lactoglobulin (~lg) adsorption kinetics on silicon surfaces varying in wettability. With regard to ~-lg, a model based on the mechanism described the data very well, enabling interpretation of the kinetic behavior in terms of contact surface hydrophobicity influences on rate constants affecting protein attachment and unfolding at the interface. In particular, both experimental and simulation results indicated that if the process of protein adsorption is resolved into two steps, the first being reversible adsorption defined by kinetic rate constants k1 and k_1 for attachment and detachment, the second being a conformational change defined by a kinetic rate constant s1 (resulting in conversion of reversibly adsorbed protein to an irreversibly adsorbed species), k1 and s1 increase with increasing solid surface hydrophobicity, while k_1 decreases. Quantitative consideration of possible mass transfer influences on the observed adsorption rates indicated that the experiments were not conducted in a transport-limited regime. In the present work, a-lac, ~-casein, and BSA adsorption kinetics were measured and interpreted with reference to the same model. To date, we have performed a number of kinetic experiments with each of the four proteins,

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but quantitative conclusions are still tentative. However, although molecularly dissimilar in several ways, differences in surface activity among these proteins at hydrophilic and hydrophobic interfaces can be at least qualitatively explained wjth reference to molecular flexibility and stability. These have been observed to be very important factors influencing a-lac, f)-casein, J3-lg and BSA interfacial behavior at air-water interfaces, and a-lac, J3-lg and BSA interfacial behavior at solid-water interfaces. f)-Casein has a largely unordered structure, but is distinctly amphiphilic; it adsorbed at a greater initial rate and in a greater amount than the other proteins on each type of surface. Results-to-date lead us to infer that it exhibits a higher affinity for hydrophilic surfaces. The globular proteins a-lac and BSA behaved in a manner more similar to that of J3-lg at each surface; however, the difference in a-lac adsorption to hydrophilic and hydrophobic surfaces was quite large, with a-lac adsorbing to a much greater extent on hydrophobic surfaces. a-Lac, a small and resilient protein, preliminarily appears to exhibit a higher initial adsorption rate, than does BSA, to hydrophobic surfaces as well. BSA consists of three large domains and nine subdomains. Its surface activity appears largely governed by relatively slow unfolding of one of these domains after adsorption. BSA did not appear to attain a plateau in adsorbed amount after eight hours of contact on either surface, although it did yield a greater adsorbed amount than a-lac in the same time period.

Impact of Research:

The ability of proteins to adsorb to food contact surfaces is well recognized. It is possible to capitalize on this phenomenon by specifically allowing antimicrobial or otherwise surface-passivating proteins to adsorb and provide an active deterrent to bacterial adhesion. Antimicrobial proteins are particularly attractive to use because of their known chemical and physical properties, their record of safe and efficacious use and their demonstrated lethality toward L. monocytogenes. The wide spectrum of food contact surfaces present in commercial settings presents a challenge to development of generic strategies that prevent adhesion and biofilm formation. Our approach includes quantitative evaluation of protein surface behavior as a function of molecular properties and surface hydrophobicity. Past research supports the importance of both hydrophobic interaction and adsorption competition on formation of an interfacial film. This work will go a long way toward allowing us to optimize noncovalent immobilization of passivating components in order to provide an effective and easily implemented barrier to Listeria adhesion.

Publications:

Daeschel, M.A., McGuire, J. and AI-Makhlafi, H. Antimicrobial activity of nisin adsorbed to hydrophilic and hydrophobic surfaces. J. Food Prot., in press.

Krisdhasima, V., McGuire, J. and Sproull, R. Surface hydrophobic influences on f)-lactoglobulin adsorption kinetics. J. Colloid Interface Sci., in press.

Suttiprasit, P., Krisdhasima, V. and McGuire, J. The surface activity of a-lactalbumin, f)-lactoglobulin and bovine serum albumin I. Surface tension measurements with single component and mixed solutions. J. Colloid Interface Sci., in press.

Suttiprasit, P. and J. McGuire. The surface activity of a-lactalbumin, f)-lactoglobulin and bovine serum albumin II. Some molecularinfluences on adsorption to hydrophilic and hydrophobic silicon surfaces. J. Colloid Interface Sci., in press .

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Project Title:

Personnel:

Funding:

Objectives:

Characterization of Milk Proteolysis by Lactococcal Starter Culture Strains Using Amino Acid Analysis

Rodney J. Brown, Dept. of Nutrition and Food Sciences, Utah State University.

Christina Beer, Dept. of Nutrition and Food Sciences, Utah State University.


1. Genetically construct lactococcal strains with different proteolytic (PrtP and PrtA) and lactose utilization capabilities.

2. Use amino acid analysis to study the interaction of milk proteolysis genes (PrtP and PrtA) of the genetically constructed strains.

3. Characterize growth and acid production of the genetically constructed strains and determine the effects that maintenance of the plasmids have on growth and acid production.

4. Examine the effect of different expression levels of proteinase genes (PrtP and · PrtA) and to try and improve flavor and texture characteristics in cheese, and examine the potential of these results for application in accelerated cheese ripening .

Results:

Twelve of the thirteen Lactococcus lactis ssp. lactis strains have been constructed. The last strain is under construction at this very moment. Amino acid analysis is being used to characterize the individual amino acid patterns when growing the cultures in different milk protein solutions. Cluster analysis for differentiating strains beyond what was possible by visual comparison of the amino acid analysis is the result.

Impact of Research:

Proteolysis from the bacterial starter cultures plays a significant role in the physical and organoleptic properties of cheese and other fermented dairy products. Improper proteolysis can result in a wide number of defects, including bitterness, texture, and body problems. We have ways to measure gross proteolysis but are very limited in techniques to profile or characterize proteolysis for individual bacterial strains. The proteinase system in Lactococci is very complex and phenotypic changes caused by genetic manipulation of a culture are impossible to measure with present proteolysis tests. This method would allow us to measure subtle differences in proteolysis as the genetic compliment of a culture is modified. An understanding of the interactions of the various genetic factors would allow for the development and optimization of cultures used in dairy fermentations. This project has the potential to enhance product quality, allow for the production of products with enhanced properties, and even allow the , development of new products by using bacterial strains with different proteolytic abilities.

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Project Title:

Personnel:

Funding Sources:

Objectives:

Final Report

Prediction and Determination of the Efficacy of Nisin in Dairy Foods

Mark A. Daeschel, Principal Investigator, Dept. of Food Science and Technology, Oregon State University

Floyd W. Bodyfelt, Principal Investigator, Dept of Food Science and Technology, Oregon State University

Dong-Sun Jung, Graduate Research Assistant, Dept of Food Science and Technology, Oregon State University

1. To determine what molecular components of milk can interact with nisin and affect its activity.

2. · To determine the efficacy of nisin on the inhibition of Listeria monocytogenes in selected

3.

milk and milk products and the subsequent impact on safety .

Documentation of how food grade emulsifiers enhance nisin activity against Listeria monocytogenes in high fat containing fluid milk.

Results:

This project provided information on the effect of dairy proteins on nisin activity. Betalactoglobulin was observed to provide a protective effect to bioassay indicator bacteria when exposed to nisin in the presence of the protein. It is hypothesized that Beta-lactoglobulin may reduce the activity of nisin by binding it and hence preventing it from inhibiting microbial cells.

More experiments were conducted to determine the effects that various components of milk have on the efficacy of nisin. Two approaches were used. 1) Determination of nisin activity after exposure of nisin to different types and concentration of dairy proteins and fats. A quantitative bioassay based on well diffusion was employed. 2) Effect of different dairy components on the ability of nisin to inhibit Listeria monocytogenes in fluid milk. The most significant effect observed was the reduction in nisin activity as the fat concentration was increased in fluid milk. Nisin activity as determined by bioassay was decreased by more than 90°/0 when added to high fat (11.5%) fluid milk. Concurrently, it was observed that nisin was less effective in inhibiting Listeria as fat concentration increased. A representative experiment is portrayed in the following figure. (Fig. 1)

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- 8 ca .... Cl) -(t) - 6 ..J - 50 U/ml Nisin --E 4 SO U/ml Nisin with Tween 80 -::J u. 0 2 C)

0 ..J 0

0 2 4 6 8 1 0 1 2 1 4 % Fat Content

Figure 1. Effect of % milkfat content of fluid milk on the ability of nisin to inhibit L. monocytogenes. Sterile milks containing 0, 10, and 50 U/ml nisin were inoculated with L. monocytogenes Jalisco at levels [log10 7.15 cfu/ml for 0 &/ml, 7.57 for 1 U/ml and 7.25 for 50 U/ml].

An additional project objective; "Documentation of how food grade emulsifiers enhance nisin activity against Listeria monocytogenes in high fat containing fluid milk" was formed on our experimental observation that the emulsifier Tween 80 appeared to "restore" nisin activity in milks containing high amounts of milkfat (4-13%) Fig. 2 and Table 1. Our working hypothesis to explain this observation is: We believe that nisin can bind to the surface of milkfat globules and when this occurs, nisin is no longer active or available to inhibit bacteria. The higher the fat content of milk, the less active or available to inhibit bacteria. The higher the fat content of milk, the less active nisin is. We believe the addition of the emulsifier Tween 80 to milk acts to displace proteins (nisin) from the milkfat globule surface resulting in a restoration of nisin activity.

The importance or implications of these observations to the dairy industry are both relevant and practical. Nisin has been approved as a GRAS additive in cheese spreads and will likely be approved for use in other dairy products. An understanding of how nisin interacts with dairy food components is paramount in order to optimize its efficacy and application in dairy foods.

Experiments were conducted to explore our hypothesis by documenting the adsorption or binding of nisin to milkfat globule and its subsequent desorption. Experimentally, we labelled the nisin molecule with a fluorescent probe and microscopically visualized its adsorption to milkfat globules. Desorption of nisin by Tween 80 was documented in the same manner. The date obtained was of a qualitative nature based on relative fluorescence and suggested that nisin does adsorb to the milkfat globule surface and Tween 80 disrupts the adsorption event.

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• TABLE 1. Effect of emulsifiers on nisin activity in half-and-half cream and skim milk

Sample treatment Inhibition zone dia. {nnn) % nisin activity (all contain 50 U/ml nisin)

skim milk 24.0 82.7 skim milk+ Tween 80 24.8 100 skim milk+lecithin 23.8 78.9 half+half cream (h+h) 16.2 19.6 (h+h)+ Tween 80 21.0 43.4 (h+h)+ lecithin 16.0 19.1 nisin 50 U/ml 24.8 100

(controls) do not contain nisin

skim milk 0 n/a (h+h) 0 n/a Tween80 0 n/a lecithin 0 n/a

n/a=not applicable

·-as 8 ... C1) -fh - 6 ..J - --a-- Nisin 0 U/ml E 4 Nisin 1 0 U/ml -:::> u.. • Nisin 50 U/ml ()

2 0)

0 ..J 0

0 2 4 6 8 1 0 1 2 1 4

o/o Fat Content

Figure 2. Effect of Tween 80 (0.2% vol/vol) on the efficacy of nisin (50 U/ml) in milks ..41111lith varying fat contentSterile milks were inoculated with L. nwnocyrogenes Jalisco at levels ~g10 7.20 cfu/ml for treaunents without Tween 80 and logw 7.60 cfu/ml with Tween 80].

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Impact of Research:

Nisin is a member of a group of potent antibacterial substances which are bacteriocins. It has been shown to be effective in inhibiting certain gram positive species but not gram negative bacteria, yeasts or fungi. Recent investigations has indicated that nisin and/or nisin producing streptococci are inhibitory toward Listeria monocytogenes, a food-borne pathogen of emerging

concern.

Nisin, after 25 years of safe use in many European countries was recently affirmed by the Food and Drug Administration (Federal Register, April 6, 1988) as GRAS for use as an antimicrobial agent to inhibit the outgrowth of Clostridium botulinum spores and toxin formation in certain pasteurized cheese spreads. The approval of nisin will justify increased research efforts of both an applied and basic nature on the antimicrobial properties of bacteriocins. It is this author's opinion that nisin will eventually be approved for use in other types of dairy foods once a sufficient body of scientific knowledge has been accumulated to justify approval. The use of nisin as an antimicrobial agent in dairy foods could enhance milk utilization by at least three

mechanisms:

1. Inhibition of spoilage microorganisms in dairy foods could minimize economic losses due to spoilage.

2.

3.

· Inhibition of pathogenic and toxigenic bacteria to provide consistently safe products. Contaminated products (such as with Listeria) can give rise to adverse publicity with subsequent sales loss .

Extension of the shelf-life of perishable dairy products.

Publications:

Daeschel, M. A., D. S. Jung and F. W. Bodyfelt. 1990. Influence of fat on the efficacy of nisin in inhibiting Listeria monocytogenes in fluid milk. Abstract in J. Dairy Sci. 73(6):Suppl. .

1.

Jung, D. S., F. W. Bodyfelt and M. A. Daeschel. 1992. Influence of fat and emulsifiers on the efficacy of nisin in inhibiting Listeria monocytogenes in fluid milk. J. Dairy Sci. 75:387-393.

Jung, D. S. Interactions and applications of nisin in food and beverage processing. Ph.D. dissertation. Dept. of Food Science and Tech., Oregon State University. (In preparation)

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Project Title: Development of a process for production of UF milk retentate

Personnel:

Funding:

Objectives:

Conly L. Hansen, Professor, Dept. of Nutrition and Food Sciences, Utah State University

Donald J. McMahon, Professor, Dept. of Nutrition and Food Science, Utah State University

Yehia A. El-Samragy, Visiting Professor, Dept. of Nutrition and Food Sciences, Utah State University

Western Dairy Foods Research Center and Utah Agricultural Experimental Station

1. Determining effects of heat treatment, pH and limited enzyme treatment of UF retentate on the chemical, physical and functional properties of its resultant spray dried, high protein, milk powder.

2. Determining effects of drying parameters, such as particle size, air temperature, solid concentration and foam spray, on properties of the retentate powder.

3. Evaluating product applications of the high protein milk powder.

Results:

ADSA abstract June 1992

Production of ultrafiltered skim milk retentate powder. 1. Composition and physical properties. Y. A. El-Samragy*, c. L. Hansen and D. J. McMahon. Utah State University, Department of Nutrition and Food Sciences, Logan 84322-8700

Raw skim milk retentate with 20% solids produced by UF was subjected to different heat treatments and pH adjustments prior to spray drying. The heat treatments were 65'c for 30 min, 75'c for 28 s, and 8s·c for 28 s. pH was adjusted to 6.4, 6.7, and 7.0. Retentate powders were analyzed for moisture, protein, lactose, fat, ash, titratable acidity, and pH. Physical property determinations included solubility index, dispersibility, viscosity, scorched particles, poured density, packed density, and water absorption isotherm. No interaction effects of heat treatment and pH adjustment were observed. Analysis of variance showed that pH adjustments had a significant effect on ash content (P < .01). pH adjustment affected only solution viscosity (P < .001) but heat treatment affected solubility and poured density (P < .05). pH adjustments and heat treatment had only minor effects on the measured properties. Results showed that by using UF to concentrate raw skim milk to 20% total solids,

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retentate powders with >60% protein can be produced which is nearly double the protein concentration of conventionally produced skim milk powder.

IFTEC abstract November 1992

Production of ultrafiltered skim milk retentate powder. 2. Functional properties-- Y. A. El-Samragy, c. L. Hansen & D. J. McMahon, Dept. of Nutr. & Food Sci. , Utah State Uni v. , Logan, Utah 84322-8700, USA

Skim milk retentate powder with modified functional properties was produced using UF and spray drying processes. Pre-treatments of different heat treatments and pH adjustments were applied prior to spray drying. The heat treatments were 65·c for 30 min, 75•c for 28 s, and 85·c for 28 s. pH was adjusted to 6.4, 6.7, and 7.0. The functional properties of the retentate powders were determined. Analysis of variance showed that pH adjustments had a significant effect on water holding capacity, acid gel strength (P < .001), emulsifying capacity and foaming capacity (P < .05). Heat treatment also significantly affected water holding capacity, acid gel strength, and emulsifying capacity (P < .001), but did not significantly affect foaming capacity. The interaction of heat treatment and pH adjustment only significantly affected foaming capacity at 5 and 10 min (P < .05) and at 15 min (P < .01) of whipping time. High protein skim milk powders with modified functional properties could be produced by using UF to concentrate raw skim milk as well as applying heat treatments and pH adjustment prior to spray drying.

Significance:

Results showed that high protein retentate powders with modified functional properties could be produced by using UF to concentrate skim milk as well as applying heat treatments and pH adjustment prior to spray drying. One more advantage for retentate powders compared with other milk powders is that the retentate powders permit desirable usage in many food systems such as yogurt, cheese, ice cream and bakery products at more economic transportation costs.

Publications:

1. Bond, s. A. and c. L. Hansen. 1991. Whippability of enzyme treated skim milk retentate powder. J. Dairy Sci. 74 (suppl. 1):95.

2. El-Samragy, Y. A., c. L. Hansen, and D. J. McMahon. 1992. Produc~i~n of ultr~filter~q ~~im m+lk .reten~ate powder. 1. Compos1t1on and phys1cal prope~~es. J. Da1ry Sc1. In Proces~.

3. El-Samragy, Y. A., C. L. Hansen, and D. J. McMahon. 1992. Production of ultrafiltered skim milk retentate powder. 2. Functional properties • J. Dairy ic.i. Hl rrocess •

r· r r' T '

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Project Title:

Personnel:

Funding:

Objectives:

High Yield, Low Moisture Cheese from Homogenized Milk

Donald J. McMahon Brian Orme Nutrition & Food Sciences Dept., Utah State University

Western Dairy Foods Research Center Utah Agricultural Experiment Station

Ultrafiltration (UF) of milk can be used for the manufacture of high moisture cheeses. Its economics lay mainly in increased retention of fat and protein. There are, however, some difficulties and complexities of making low moisture cheese using UF. High fat losses occur and it is difficult to remove moisture. The objectives of this project are:

1. Determine effects of homogenization treatment on fat losses from UF retentate curd.

2 .. Design a cheese making process so as to obtain cheese in the range of pH 5.0-5.4 and moisture content <40%.

3. Determine effects of milk heat treatment on moisture, texture and body of cheese made from UF retentate.

4. Provide a manufacturing procedure for making acceptable low moisture cheese from pre-fermented UF retentate that could be adopted for a continuous cheese making process.

Results:

The extent of fat-protein complexing was determined over a range of homogenization temperatures and pressures. Higher pressures produced more fat-protein complexing but resulted in poorer quality texture of cheese made from homogenized UF retentate. Coagulation trials were conducted to determine the best temperature and pH conditions for adding rennet to UF retentate. The relatively high viscosity of 5X UF retentate makes the even addition of rennet difficult. This can be overcome by lowering retentate pH, however, it was decided that a better approach to overcoming processing difficulties was to use a 4X UF retentate. Additional advantages gained by doing so, was that diafiltration was no longer required and the cheese curd could be cut using conventional cheese harps. Microbial growth rates of starter culture strains in 4X UF retentate were compared to their growth rates in milk and appropriate pairs of strains were , determined.

After much refining of the cheesemaking process, trials were conducted using 34% solids UF retentate at two homogenization temperatures, 40°C and 60°C and compared to a nonhomogenized control from the same batch of retentate.

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Homogenization pressure of BOO psi first stage and 400 psi second stage were used at both temperatures. Cheese curd was then made by renneting the retentate and adding a 55% overlay of water before cutting the curd. All samples had a pH of 5.05-5.15 after 1 day and the target moisture of <40% was obtained. The retentate homogenized at 60°C gave the highest yield but produced a cheese with a short texture. The nonhomogenized retentate cheese was also short and was crumbly. It had a much lower yield and there was excessive "oiling off" as typically occurs when renneting UF retentate. The retentate homogenized at 40°C produced a cheese with the best texture and gave a good yield.

Impact of Research:

The technology of making cheese by UF has been most successful in the production of high moisture cheeses in Europe. UF has been of only limited use in the manufacture of low moisture cheeses typically produced in the United States. Commercial systems utilizing UF have been capital-intensive and although providing a continuous operation have not shown the expected yield increase. Through the application of the results of this research it should be possible to make UF cheeses with increased yields and in less capital-intensive systems. The procedures developed could even be adapted to use in conventional cheese vats. Applying the results of the microbial growth studies would also help prevent strain dominance and thus reduce phage problems.

Publications: Orme, B.J. and D.J. McMahon. 1989. Effect of concentration on coagulation of

ultrafiltered milk. 84th American Dairy Science Association Meeting, J. Dairy Sci:Supp. 1, 135.

McMahon, D.J. and R.J. Brown. 1990. Development of surface functionality of casein particles as the controlling parameter of enzymic milk coagulation. Colloids and Surfaces. 44:263-279.

Orme, B.J., D.J. McMahon and R.K. Thunell. Variable growth and acid production by Lactococcus /actis strains in ultrafiltered concentrated whole milk. J. Dairy Sci. (Submitted) .

63

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Project Title:

Personnel:

Funding:

Use of Ultrafiltration and Different Heat Treatments on Yogurt Flavor and Physical Properties

Paul A. Saye!lo. Dept. of Nutrition and Food Sciences, Utah State University

Richard A. Dargan, Dept. of Nutriton and Food Sciences, Utah State University


Project Period: August 1, 1988 to September 30, 1991

Objectives:

1. To measure the effects on the yogurt properties of viscosity, gel strength, syneresis, and water holding capacity by ultrafiltering yogurt milk to different total milk solids levels and applying different heat treatments;

2. To measure the whey protein denaturation in the yogurt milk as a result of the different heat treatments;

3. To observe structural differences by scanning electron microscopy and transmission electron microscopy of the different-treated yogurt milks;

4. To measure the acceptability of yogurt flavor and body/texture by appropriate taste panel procedures;

5. To measure the effect on acidification time to desired gelation level by different heat treatments.

Results:

Objective 1: To measure the effects on the yogurt properties of viscosity, gel strength, syneresis, and water holding capacity (WHC) by ultrafiltering to different total milk solids levels and applying different heat treatments.

Materials and Methods: Skim milk, 1% fat milk, skim milk ultrafiltered to 12.5% solids, and skim milk ultrafiltered to 15% solids were ultra-high temperature (UHT) treated by means of an Alfa-Laval Sterilab to 220, 240, 260, or 28o·F for 4 sec. All milks except the 1% fat milk were also UHT treated for 8, 12, or 16 sec at each of the above temperatures. Control yogurts were prepared by vat pasteurization of aliquots of the same milks. Additional controls were prepared by vat pasteurization of skim milk with 2, 3, 4, or 5% added nonfat dry milk (NFDM) as a ''traditional process" comparison. Vat heat treatment was 182.F for 20 minutes. Forty minute vat heated samples were also prepared for 2, 3, 4, or 5% added NFDM and 12.5% UF solids skim milks .

64

Measurements of viscosity, gel strength, syneresis, and water holding capacity • were made on samples from all treatments at 7, 14, and 21 days.

•

Viscosity. Stirred viscosity measurements were made by Brookfield viscometer at 3 rpm with T-bar spindles. Following 30 seconds stirring, readings were taken every 30 seconds. An average of 4 readings was calculated for each sample. Four samples were tested per treatment.

Gel strength. A device was manufactured to give a relative measure of gel strength. A 5/8 inch shaft was lowered at a fixed speed into 1 00 ml samples on top of an analytical balance. Breaking force (gel strength) was taken to be the first point at which the balance reading stops increasing and decreases sharply. Four samples were determined per treatment.

Syneresis. Syneresis was measured by aspirating the whey off of the samples when inclined at a so· angle. Four samples were measured for each treatment.

Water holding capacity. Water holding capacity (WHC) was measured by centrifuging approximately 40 grams of sample at 10,500 rpm for 30 minutes at 1 o·c. The supernatant was removed by draining the tube in an inverted position for 1 0 min. WHC was expressed as percent pellet weight relative to sample weight. Two samples were measured for each treatment.

Results: Viscosity of yogurt increased with increased UF solids (Figure 1 ). Viscosity of UHT yogurt was greater than the viscosity of yogurt from vat heated skim milk yogurt with comparable total solids from added NFDM. Viscosity did not increase directly with increased UHT temperature. Rather, viscosity was greatest from lower (220 and 240"F) temperature treatments. AT 280"F, viscosity was reduced compared to other UHT temperatures. Viscosity tended to decrease with increasing UHT temperature at higher solids levels (12.5 and 15% UF solids). Increased holding time at 280"F increased viscosity.

Gel strength increased with increasing level of skim milk solids as a result of ultrafiltration. Gel strength was higher in yogurt made from UF/UHT milks than in vat heated yogurt made with added NFDM to even higher levels of solids (Figure 2). UHT yogurt from skim milk with 1% added milkfat had lower gel strength than yogurt from skim milk alone. Gel strength did not seem to improve directly with increased UHT temperature nor did holding time dramatically increase gel strength. Temperature, however, had more impact on gel strength than did holding time. 280"F had an unfavorable impact on gel strength at all solids levels. In most cases the lower temperatures (220 and 240"F) showed greatest gel strength. Twenty minute vat heat treatment of UF milks resulted in higher gel strength than any UHT treatment.

Higher UF solids decreased syneresis by more than would be expected by increased solids alone based on comparison to vat-heated yogurt milks with added NFDM (Figure 3). Addition of 1% milkfat also reduced syneresis.

• Increased UHT temperature increased syneresis in skim milk yogurt but the

65

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•

•

effect seemed to be overcome by the increased solids in the UF samples. Increased holding time had a pronounced effect on reducing syneresis of skim milk yogurt from the 2so·F treatment, but had no effect on the already low syneresis in the higher solids yogurts (Figure 4). Syneresis was higher on vat treated yogurt than UHT yogurt.

Water holding capacity increased with increased level of skim milk solids as a result of ultrafiltration. The added solids from UF had a greater impact on increasing WHC than did comparable solids levels from addition of NFDM in a traditional (vat) heating process (see Figure 5). Yogurt made from 1% fat milk also had greater WHC than did skim milk yogurt. WHC did not increase with increasing UHT temperature. Rather, lower (220 and 240.F) temperatures seemed to provide the greatest WHC. Increased holding time did not improve WHC and, in some cases, reduced it.

Conclusions: Using ultrafiltration to increase solids affords greater viscosity, gel strength, WHC, and lower syneresis in UHT yogurt as compared to vat heated yogurt with added NFDM to comparable solids levels. Increasing UHT temperature (especially 280.F) does not improve gel strength, viscosity, WHC, and syneresis. Rather, lower temperatures (220 and 240.F) provide the greatest viscosity, gel strength, WHC, and least syneresis (especially when the solids are increased by UF). Intermediate UHT treatment may provide comparable or better WHC, and improved syneresis as compared to vat-heated yogurt, but this effect becomes less distinguishable as solids are increased by UF (both heating methods are good). However, UHT treatment does not appear to be able to match the same levels of gel strength and viscosity that can be seen with vat heat treatment.

Objective 2: To observe structural differences by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) of the differently treated yogurt milks.

Electron microscopy (EM) of samples has been completed by Chico State University Electron Microscopy Facility. Specialized antibody (for K-casein and 13-lactoglobulin) and staining techniques were used to locate these milk proteins in the yogurt. The EM prints are being reviewed for future manuscript preparation

Objective 3: To measure acceptability of yogurt flavor and texture by appropriate taste panel procedures.

Taste and sensory panels were not performed.

Objective 4: To measure the effect on acidification time to desired gelation level by different heat treatments.

No significant differences were measured in acidification times as a result of different heat treatments .

66

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•

Objective 5: To measure whey protein denaturation in the yogurt milk as a result of the different heat treatments .

Materials and Methods: Yogurt milks were prepared as in Objective 1. Unfermented samples of all heat treatments were tested for whey protein denaturation (WPD). Milk samples were adjusted to pH 4.6 by addition of 0.1 N HCI. The precipitated casein was filtered out and the whey-protein-containing filtrate was collected. Kjeldahl nitrogen analysis was performed on the filtrate samples. The whey protein level found in the heat treated samples was compared to the whey protein level found in the raw milk of the same concentration. Whey protein denaturation was calculated as the disappearance of the whey protein in heat treated samples as compared to unheated controls as a result of whey protein-casein complexation.

Besylts: Yogurt milks with higher solids from UF had higher levels of WPD at each heat treatment (Figure 6). WPD increased with increasing UHT temperature and holding time. The effect of temperature was more dramatic at lower holding time (Figure 7). Temperature had a more dramatic effect on WPD than did holding time (except for holding time at 220.F). Vat heat treatment resulted in WPD that was at or near the same level achieved by the most severe UHT treatments. It is interesting to note, however, that the highest WPD in UHT treatments did not correlate with the best yogurt properties (see Objective 1 Results). In fact, inferior viscosity, gel strength, WHC, and syneresis resulted at 2ao·F .

Conclusions: Whey protein denaturation may not be a good indicator of UHT yogurt properties. Although UHT treatments involve high temperatures, the resulting WPD is not as great as seen with lower temperature, longer time vat heating methods.

Impact of Research: This research indicates combining the technologies of ultrafiltration and ultra-high temperature treatment of yogurt milk can produce comparable or superior yogurt than traditional (vat heat treatment) yogurt of comparable total solids levels. The research has thus far indicated that the highest UHT temperatures of yogurt milk do not yield improved product characteristics over "lower" UHT temperatures.

Increasing the total milk solids by ultrafiltration appears to be a good means for improved yogurt physical characteristics. Measurements of these yogurt flavor and texture attributes must still be studied .

67

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APPENDIX A

WDFRC BALANCE SHEET FOR FV88 TO FV92

WDFRC Proposed Project Allocations

WDFRC Actual Project Allocations

Total WDFRC Funds Spent

Balance (Actual Allocation -Funds Spent)

Returned (Unspent) Project Funds

Overspent Project Funds

FY88 to FY92 Allocated Project Funds To Be Used For Project Extensions Into FY93

TOTAL BALANCE FORWARD TO FY93

$2,749,790

2,753,449

2,436,488

$316,961

25,577

(817)

(263,803)

$77,918

•

•

•

APPENDIX 8

NEW WCDPRT PROJECTS 1992-1993

Purification of monospecific polyclonal antibodies from bovine cheese whey. Rodney J. Brown, Utah State University.

Bacteriophage-resistance gene replacement in Lactococcus lactis. Bruce Geller, Oregon State University.

Production of extracellular proteases of Brevibacterium linens for use in lowfat cheese. Bart Weimer, Utah State University.

Interactions between milk proteins, starter cultures, and hydrocolloidal milk fat replacers. Bart Weimer, Utah State University.

Effects of iron fortification on chemical, physical, microbiological and nutritional properties of yogurt. Donald J. McMahon, Utah State University.

Effects of whey proteins and lactose in age gelation of UHT-processed milk concentrate- Part 2. Donald J. McMahon, Utah State University.

Using a natural process to improve milk quality and extend milk shelf-life through the reduction in lipid oxidation and off-flavors with tocopherol (Vitamin E) supplementation to dairy cows. Gerald Schelling, University of Idaho.

Rheology and microstructure of Mozzarella cheese. Donald J. McMahon, Utah State University.

Extrusion processing of milk proteins. Conly L. Hansen, Utah State University .

•

APPENDIXC

ANNUAL FINANCIAL REPORTS BY PROJECT

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t UTAH STATE U N I V E R S I T Y • L 0 G A N, U T A H 8 4 3 2 2

OFFICE OF INTERNAL AUDITS UMC 1430 Telephone (801) 750-1084

To National Dairy Promotion and Research Board

October 8, 1992

QUALIFIED INTERNAL AUDITORS' STATEMENT

We have performed an accounting of the Annual Financial Report by Project, of

the Western Dairy Research Center, from July 1, 1987 to June 30, 1992 .

Utah State University October 8, 1992

• • • ANNUAL FINANCIAL REPORT BY PROJECT

Weste~~ Da±ry Research Ce~te~

Annual Report Ending: June 30, 1992, Final Project Term: 07/01/87 - 06/30/89

Dairy Center Director: Dr. Paul A. Savello Principal Investigator: HANSEN CONLY L

Project Title: COGENERATION OF BIOGAS & SINGLE CELL PROTEIN FROM ULTRAFILTRATION PERMEATE & WHEY

Project Status: Closed USU Project Number: 189 USU Account Number: 547766

==================================================================================================I I Budget I I Total Funds I I NDPRB Funds I I Local Funds I I

Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========1 1========1========1=========1 1========1========1=========1 1========1========1=========1 I salaries/ II I I II I I II I I I I Wages I I 50774 50774 0 16923 16923 0 33851 33851 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I I 2435 2435 0 812 812 0 1623 1623 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I I 4897 4963 -66 1632 1654 -22 3265 3309 -44 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I I 0 0 0 0 0 0 0 0 0 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel I I 966 966 0 322 322 0 644 644 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! I 106 40 66 35 13 22 71 27 44 ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I II I I I I I I II TOTALS I I 59178 59178 0 19724 19724 0 39454 39454 0 ==================================================================================================1 I


Wester~ Dairy Research Ce~ter


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: KONDO JEFFERY K

Project Title: CLONING THE NISIN & OTHER GENES OF LACTIC STREPTOCOCCI INTO LEUCONOSTOC SPECIES & AMI LIFICATION OF NISIN PRODUCTION

Project Status: Active USU Project Number: 188 USU Account Number: 547767

==================================================================================================1 I Budget I I Total Funds I I NDPRB Funds I I Local Funds I I

Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========1 1========1========1=========1 1========1========1=========1 1========1========1=========1 I Salaries; I I I I I I I I I I I I I I

Wages I I 88484 99084 -10600 63499 71105 -7607 24985 27979 -2993 -----------11--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I I 29442 10336 19106 21128 7417 13711 8314 2918 5395 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I I 45846 57719 -11873 32900 41421 -8520 12946 16298 -3353 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I I 12560 4321 8239 0 0 0 12560 4321 8239 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I I 1000 1382 -382 718 992 -274 282 390 -108 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! I 0 835 -835 0 599 -599 0 236 -236 ===========!I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

II I I II I I II I I II TOTALS I I 177332 173677 3655 118245 121534 -3289 59087 52142 6945 ==================================================================================================I I


Weste~~ DaXry Rese~ch Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: MAHONEY ARTHUR W

Project Title: IRON FORTICIATION OF CHEESE CURD


=========== Budget

Summary =========== Salaries/ Wages

Fringe

Supplies

Equipment

Travel

Publication! ===========!

I TOTALS I ============

======================================================================================!I I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I

========J========J=========I J========J========J=========J I========J========J=========J I I I II I I II I I II

35000 35000 0 23331 23331 0 11669 11669 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

4599 4599 0 3065 3065 0 1533 1533 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

2504 2504 0 1669 1669 0 835 835 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

100 100 0 67 67 0 33 33 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 42203 42203 0 28133 28133 0 14071 14071 0

=====================================================================================!I


Waste~~ DaXry Rese~ch Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: RICHARDSON GARY H

Project Title: ACQUISITION OF ZYMARK II ROBOT FOR LABORATORY AUTOMATION STUDIES


======~==== ======================================================================================I I Budget I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent I Balance IAllocatedJ Spent I Balance JAllocatedl Spent I Balance I I =========== 1========1========1=========1 1========1========1=========1 J========I========J=========I I Salaries/ I I I I I I I I I I I I I Wages I 1474 1474 0 983 983 0 491 491 0 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I 0 0 0 0 0 0 0 0 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I 1455 1446 9 970 964 6 485 482 3 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I 0 0 0 0 0 0 0 0 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 1963 1963 0 1309 1309 0 654 654 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! I 108 117 -9 72 78 -6 36 39 -3 ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

II I I II I I II I I II TOTALS I I 5000 5000 0 3334 3334 0 1667 1667 0 ==================================================================================================!I



Dairy Center Director: Dr. Paul A. Savello Principal Investigator: SAVELLO PAUL A

Project Title: DAIRY CENTER GENERAL EXPENSES FROM NDPRB FUNDS


================a===================================================•=============================J J

Budget I I Total Funds I I NDPRB Funds I I Local Funds I I Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent 1 Balance 1 1

===========I ========I======== I========= I I =••===== I======== I =======•= I I•======= I======== I========= I I Salaries/ I I I I I I I Wages I 0 0 0 0 0 0 0 0 0 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I 0 0 0 0 0 0 0 0 0 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I S upp 1 i e s I 0 0 0 0 0 0 0 0 o -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equ i pmert t I 0 0 0 0 0 0 0 0 0 ~----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 0 0 0 0 0 0 0 0 0 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! 27 27 0 27 27 0 0 0 o ===========I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I TOTALS 27 27 0 27 27 0 0 0 0 ====================~==•==•=========•==c===•=======================•=====•========================J I


Waste~~ Dairy Rese~ch Ce~te~



Project Title: DAIRY CENTER GENERAL EXPENSES FROM NON-NDPRB FUNDS


===========-======================================================================================!I Budget I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I =========== ========I========!========= I 1========1========1=========1 1========1========1=========1 I Salaries/ I I I I I I II I I I I Wages 18000 18000 0 0 0 0 18000 18000 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe 6395 6395 0 0 0 0 6395 6395 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 18135 18135 0 0 0 0 18135 18135 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 25000 25000 0 0 0 0 25000 25000 0 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel 0 0 0 0 0 0 0 0 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication 0 0 0 0 0 0 0 0 0 =========== -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I TOTALS 67529 67529 0 0 0 0 67529 67529 0 ==================================================================================================!I





Project Title: IMPROVING YIELD & PHYSICAL PROPERTIES OF MOZZARELLA CHEESE


Budget Summary

===========-======================================================================================I I I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========

Salaries/ Wages

Fringe

Supplies

Equipment

Travel

Publication ===========!

I TOTALS I ============

========1========1=========1 1========1========1=========1 1========1========1=========1 I I I II I I II I I II

48409 48409 0 32509 32509 0 15900 15900 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

13122 13122 0 8812 8812 0 4310 4310 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

5405 5405 0 3630 3630 0 1775 1775 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

400 400 0 0 0 0 400 400 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

435 435 0 292 292 0 143 143 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

197 197 0 132 132 0 65 65 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 67968 67968 0 45375 45375 0 22593 22593 0

=====================================================================================!I


Waste~~ D~y Research Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: ERNSTROM C ANTHON

Project Title: CONTINUOUS PRODUCTION OF COTTAGE CHEESE FROM ULTRA-FILTRATED SKIM MILK RETENATE



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========II======== I======== I========= II======== I======== I========= II======== I======== I========= I I Salaries/ II I I I I I I I I I I I I Wages I I 1950 1950 0 1950 1950 0 0 0 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I I 637 637 0 637 637 0 0 0 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I 5631 5631 0 5631 5631 0 0 0 0 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 1 0 o 0 0 0 0 0 0 0 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 435 435 0 435 435 0 0 0 0 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! 47 47 0 47 47 0 0 0 0 ===========! -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I 8700 8700 0 8700 8700 0 0 0 0 ============ =====================================================================================I I


Western.. Da.i.:ry Resea.:cc::h Cen..t.er


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: CORNFORTH DAREN P

Project Title: EVALUATION OF MILK PROTEINS AS WHITENING AGENTS IN PROCESSED MEATS & POULTRY PRODUCT:


==================================================================================================!I Budget I I Total Funds I I NDPRB Funds I I Local Funds I I

Summary !Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I ===========I I======== I======== I========= I I======== I=====::::== I========= I I======== I======== I========= II salaries/ II I I II I I II I I II Wages I I 24671 24671 0 16723 16723 0 7948 7948 0 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I I 1004 1004 0 680 680 0 323 323 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 1 I 5675 5675 0 3847 3847 0 1828 1828 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipm0nt I I 544 544 0 0 0 0 544 544 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I I 978 978 0 663 663 0 315 315 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication!! 201 201 0 136 136 0 65 65 0 ===========!I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I





Dairy Center Director: Dr. Paul A. Savello Principal Investigator: OLSEN ROBERT L

Project Title: INTERACTION OF PROTEIN & POLYSACCHARIDES IN CHYMOSIN & ACID COAGULATION OF MILK



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance 1 1 ===========I 1========1========1=========1 1========1========1=========1 1========1========1=========1 I Salaries/ I I I I II I I I I I I I I Wages I I 25652 25652 0 17670 17670 0 7982 7982 0 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I I 5714 5714 0 3936 3936 0 1778 1778 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I I 8538 8538 0 5881 5881 0 2657 2657 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I I 1352 1352 0 0 0 0 1352 1352 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I I 800 800 0 551 551 0 249 249 0 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! I 0 0 0 0 0 0 0 0 0 ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I






Project Title: IMPROVED CONTROL OF CHEESE MANUFACTURE THROUGH VAT MONITORING



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========I 1========1========1=========1 1========1========1=========1 1========1========1=========1 I Salaries/ I I I I II I I I I I I I I Wages I I 36200 34326 1874 27160 25754 1406 9040 8572 468 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I 1 9100 1069 8031 6827 802 6025 2273 267 2006 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Supplies I I 8000 24909 -16909 6002 18689 -12686 1998 6221 -4223 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I I 7000 500 6500 0 0 0 7000 500 6500 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel I I 3000 1850 1150 2251 1388 863 749 462 287 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! I 0 533 -533 0 400 -400 0 133 -133 ===========!I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

II I I I I I I I I I I I I TOTALS I I 63300 63186 114 42240 47032 -4792 21060 16155 4905 ==================================================================================================!I


Waste~~ D~y Rese~ch Ce~te~



Project Title: WESTERN DAIRY FOODS RESEARCH CENTER ADMINISTRATIVE ACCOUNT


==================================================================================================!I Budget I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I =========== ========1========1=========1 1========1========1=========1 I========!======== I=========! I Salaries/ I I I I I I I I I I I I Wages 85000 83714 1286 60052 59144 909 24948 24570 377 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe 28000 27305 695 19782 19291 491 8218 8014 204 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 29000 23346 5654 20488 16494 3995 8512 6852 1659 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 6000 6075 -75 0 0 0 6000 6075 -75 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel 49000 47845 1155 34618 33802 816 14382 14043 339 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication I 3000 3608 -608 2119 2549 -430 881 1059 -179 =========== I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I TOTALS I 200000 191893 8107 137060 131279 5781 62940 60613 2327 ==================================================================================================!I


Waste~~ Da~y Research Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: ERNSTROM C ANTHON

Project Title: EFFECT OF MILK CLOTTING ENZYMES ON THE CURING & QUALITY OF CHEDDAR CHEESE


=========== ======================================================================================1 I I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I

Budget Summary

=========== 1========1========1=========1 1========1========1=========1 1========1========1=========1 I I I I I I I I I I I I I I Salaries/

Wages I 27837 27837 o 18509 18509 o 9328 9328 o ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

I 541 541 o 360 360 o 181 181 o ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

I o o o o o o o o o ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

I o o o o o o o o o ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I

I 600 600 o 399 399 o 201 201 o ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication I 46 46 0 31 31 0 15 15 0 =========== I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I I TOTALS I I 29024 29024 0 19298 19298 0 9726 9726 0 ==================================================================================================!I

Fringe

Supplies

Equipment

Travel




Dairy Center Director: Dr. Paul A. Savello Principal Investigator: SANDINE W E

Project Title: CLONING THE NISIN & OTHER GENES OF LACTIC STREPTOCOCCI INTO LEUCONOSTOC SPECIES & AM: LIFICATION OF NISIN PRODUCTION



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========I ========I======== I========= II======== I======== I========= I I======== I======== I========= I I Salaries/ I I I I I I I I I I I II Wages I 129370 105384 23987 87608 71365 16243 41762 34019 7743 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I 36406 25155 11251 24654 17035 7619 11752 8120 3632 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I 61047 66298 -5251 41340 44896 -3556 19707 21401 -1695 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I 3500 3955 -455 0 0 0 3500 3955 -455 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 0 2265 -2265 0 1534 -1534 0 731 -731 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! 0 0 0 0 0 0 0 0 0 ===========I -------1 ------1 . -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I 230323 203056 27267 153602 134829 18773 76721 68226 8494 ============ =====================================================================================1 I


Weste~~ D8Xry Research Ce~te~



Project Title: CHARACTERIZATION OF BACTERIOPHAGE RECEPTOR SITES OF LACTIC STREPTOCOCCI



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I =========== ========1========1=========1 1========1========1=========1 1========1========1=========1 I Salaries/ I I I I I I I I I I I I Wages 0 27994 -27994 0 18599 -18599 0 9395 -9395 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe 0 3223 -3223 0 2142 -2142 0 1082 -1082 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Supplies 55272 21887 33386 36723 14542 22181 18549 7345 11204 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment 0 0 0 0 0 0 0 0 0

----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel 0 2169 -2169 0 1441 -1441 0 728 -728 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication 0 0 0 0 0 0 0 0 0

=========== -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I I I II I I II I I II

TOTALS 55272 55272 0 36723 36723 0 18549 18549 0 ============ =====================================================================================I I

-------------

• • • ~NNUAL FINANCIAL REPORT BY PROJECT

Waste~~ Dadry Research Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: BODYFELT FLOYD W

Project Title: RAPID ASSAY FOR HEAT RESISTANT MICROBIAL PROTEASES IN RAW MILK BY A SIMPLE CASEIN DE~ ATURATION METHOD


=========== ======================================================================================!I Budget

Summary I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I

=========== ========1========1=========1 1========1========1=========1 1========1========1=========1 I Salaries/ Wages

I I I I I I I I I I I I 14400 17623 -3223 9618 11770 -2153 4782 5853 -1070

----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe 5830 2999 2831 3894 2003 1891 1936 996 940 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 14350 13235 1115 9584 8840 745 4766 4395 370 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 0 0 0 0 0 0 0 0 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel 2000 1084 916 1336 724 612 664 360 304 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication 0 0 0 0 0 0 0 0 0 =========== I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I TOTALS I I 36580 34941 1639 24432 23337 1095 12148 11604 544 ==================================================================================================I I



Dairy Center Director: Dr. Paul A. Savello Principal Investigator: BODYFELT FLOYD W

Project Title: PRODUCTION OF OMEGA-3 FATTY ACIDS BY GENETICALLY ALTERED FUNGI & LACTIC ACID BACTERii



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========I I======== I======== I========= II======== I======== I========= II======== I======== I========= I I Salaries/ I I I I I I I I I I I I I I Wages I I 14400 21426 -7026 9556 14218 -4662 4844 7208 -2364 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I I 8705 3683 5022 5777 2444 3333 2928 1239 1689 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Supplies I I 29550 25318 4232 19609 16801 2808 9941 8517 1424 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I I 0 1791 -1791 0 0 0 0 1791 -1791 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel I I 2000 1584 416 1327 1051 276 673 533 140 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! I 0 0 0 0 0 0 0 0 0 ===========I I -------1 -~----1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

II I I II I I II I I II TOTALS I I 54655 53802 853 36269 34514 1755 18386 19288 -902 ==================================================================================================I I


Waste~~ Dairy Research Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: MCGUIRE JOSEPH

Project Title: CHARACTERIZATION OF THE POST-ABSORBTIVE BEHAVIOR OF B-LACTOGLOBULIN FOR CONTROL OF SI ORE & MICROBIAL ADHESION



Summary IAllocatedl Spent I Balance IAllocatedJ Spent I Balance IAllocatedl Spent I Balance I I ===========I ========I======== I========= II======== I======== I========= II======== I======== I========= I I Salaries/ I I I II I I II I I II Wages I 27360 33916 -6556 18268 22646 -4377 9092 11270 -2179 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I 9436 2363 7073 6300 1578 4723 3136 785 2350 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I 35800 36379 -579 23904 24290 -387 11896 12089 -192 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I 0 0 0 0 0 0 0 0 0 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 4000 3938 62 2671 2630 41 1329 1309 21 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! 0 0 0 0 0 0 0 0 0 ===========1 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I I 76596 76596 0 51143 51143 0 25453 25453 0 ==================================================================================================!I





Project Title: STUDIES ON THE GROWTH & SURVIVAL OF BIFIDOBACTERIUM SPECIES IN MILK

Project Status: Closed USU Project Number: 198 usu Account Number: 547814


Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I =========== I======== I======== I========= II======== I======== I========= II======== I======== I========= II Salaries; I I I II I I II I I II Wages I 14760 17873 -3113 9840 11916 -2075 4920 5957 -1037 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I 5923 106 5817 3949 70 3878 1974 35 1939 ----------- 1-------~1--------1---------1 1--------1--------1---------11--------1--------1---------1 I Supplies I 16000 18705 -2705 10667 12471 -1803 5333 6234 -902 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I 0 0 0 0 0 0 0 0 0

----------- 1--------1--------1---------1 1--------1--------1---------11--------1--------1---------1 I Travel I 0 0 0 0 0 0 0 0 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication I 0 0 0 0 0 0 0 0 0

=========== I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I I I I II I I II I I II

TOTALS I 36683 36683 0 24457 24457 0 12226 12226 0 ==================================================================================================1 I


Weste~~ D~y Rese~ch Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: OGDEN LYNN V

Project Title: CONTINUOUS PRODUCTION OF COTTAGE CHEESE FROM ULTRAFILTRATED SKIM MILK RETENATE


=========== ======================================================================================!I Budget I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I =========== ========I======== I========= I 1========1========1=========1 1========1========1=========1 I Salaries/ I I I I I I II I I I I Wages 6124 6124 0 4081 4081 0 2043 2043 0 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe 0 0 0 0 0 0 0 0 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 428 428 0 285 285 0 143 143 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 0 0 0 0 0 0 0 0 0 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel 1042 1042 0 694 694 0 348 348 0 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! 0 0 0 0 0 0 0 0 0 ===========! -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I I I TOTALS I 7594 7594 0 5061 5061 0 2533 2533 0 ==================================================================================================!I





Project Title: METHOD FOR IDENTIFYING BATCH OF ORIGIN OF SEMI-CONTINUOUS CHEESE PROCESSES


======~====

Budget Summary

=========== Salaries/ Wages

Fringe

Supplies

Equipment

Travel


I TOTALS I ============

======================================================================================I I I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance !Allocated! Spent I Balance IAllocatedl Spent I Balance I I

========1========1=========1 I======== I======== I========= I 1========1========1=========1 I I I II I I II I I II

6100 6100 0 0 0 0 6100 6100 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 6100 6100 0 0 0 0 6100 6100 0

=====================================================================================I I


Waste~~ DaXry Research Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: BROWN RODNEY J

Project Title: APPLICATION OF FOURIER TRANSFORM INFRARED TECHNOLOGY TO MILK & DAIRY PRODUCTS



Summary !Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I ===========I ========I======== I========= II======== I======== I========= I I======== I======== I========= II Salaries/ I I I I I I I I I I I I I Wages I 53593 45377 8216 41377 35034 6343 12216 10343 1873 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I 16078 14686 1392 12413 11338 1075 3665 3347 317 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I 3000 18387 -15387 2316 14196 -11880 684 4191 -3507 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I 50250 41015 9235 0 0 0 50250 41015 9235 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 2000 3865 -1865 1544 2984 -1440 456 881 -425 -----------1 --------1--------1---------11--------1--------1---------1 1--------1--------1---------1 I Publicationl 0 258 -258 0 199 -199 0 59 -59 ===========I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I 124921 123587 1334 57651 63752 -6100 67270 59836 7434 ============ =====================================================================================!I


Weeber~ Dairy Research Ce~ter



Project Title: ESTIMATION OF INDIVIDUAL MILK PROTEINS & GENETIC VARIANTS BY MULTICOMPONENT ANALYSIS OF AMINO ACID PROFILES


=======================================~==========================================================!I Budget I I Total Funds I I NDPRB Funds I I Local Funds I I

Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========I ========1========1=========1 1======~=1========1=========1 1========1========1=========1 I Salaries; I I I I I I I I I I I I I Wages I 59110 28833 30277 47199 23023 24176 11911 5810 6101 -----------1 --------1--------1---------1 1--------1--------1~--------1 1--------1--------1---------1 I Fringe I 17733 5270 12463 14160 4208 9952 3573 1062 2511 -----------1 --------1--------1---------11--------1--------1---------1 1--------1--------l---------1 I Supplies I 169050 147576 21474 134986 117839 17147 34064 29737 4327 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I 0 0 0 0 0 0 0 0 0 -----------1 --------1--------1---------11--------1--------1---------1 1--------1--------1---------l I Travel I 2000 1976 24 1597 1578 19 403 398 5 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! 0 259 -259 0 207 -207 0 52 -52 ===========! -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I 247893 183914 63979 197943 146855 51087 49950 37059 12892 ============ =====================================================================================!I




Dair; Center Director: Dr. Paul A. Savello Principal Investigator: SAVELLO PAUL A

Project Title: USE OF ULTRAFILTRATION & DIFFERENT HEAT TREATMENTS ON YOGURT FLAVOR & PHYSICAL PROPEF TIES


=========== ======================================================================================!I I Total Funds I I NDPRB Funds I I Local Funds I I Budget

Summary Allocated! Spent I Balance !Allocated! Spent I Balance IAllocatedl Spent 1 Balance 1 I =========== ========I========!========= I 1========1========1=========1 1========1========1=========1 I

I I I I I I I I I I I I Salaries/ Wages 32960 39325 -6365 22030 26285 -4254 10930 13040 -2111 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

8240 997 7243 5508 666 4841 2732 331 2402 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

17000 16370 630 11363 10942 421 5637 5428 209 ----------- --------1--------1---------1 1--------1-:------1---------1 1--------1--------1---------1 I

0 480 -480 0 0 0 0 480 -480 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 400 -400 0 267 -267 0 133 -133 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 628 -628 0 420 -420 0 208 -208 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II TOTALS 58200 58200 0 38901 38580 321 19299 19620 -321 ==================================================================================================1 I

Fringe

Supplies

Equipment

Travel

Publication ===========




Dairy Center Director: Dr. Paul A. Savello Principal Investigator: DAESCHEL MARK A

Project Title: PURIFICATION OF A BACTERIOCIN FROM PEDIOCOCCUS PENTOSACEUS & GENETIC TRANSFER OF THE PLASMID BORNE DETERMINANT


===========-======================================================================================!I Budget


Fringe

Supplies

Equipment

Travel

Publication I ======~====I

I TOTALS I ============

I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I

========1========1=========1 1========1========1=========1 1========1========1=========1 I I I II I I II I I II

23892 31379 -7487 15984 20993 -5009 7908 10387 -2478 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

9536 6475 3061 6380 4332 2048 3156 2143 1013 --------1--------1---------1 1--------1--------1---------1 1--------1~-------1---------1 I

17534 16112 1422 11730 10779 952 5804 5333 471 --------1--------1---------1 1--------1--------1---------11--------1--------1---------1 I

0 239 -239 0 0 0 0 239 -239 --------1--------1---------1 1--------1--------1---------11--------1--------1---------1 I

3500 257 3243 2342 172 2169 1159 85 1073 --------1--------1---------1 1--------1--------1---------1 1--------1---~----1---------1 I

0 0 0 0 0 0 0 0 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 54462 54462 0 36435 36275 160 18027 18187 -160

=====================================================================================!I


Weste~~ Da±ry Rese~ch Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: TORRES J ANTONIO

Project Title: CHEDDAR CHEESE BLOCKS: EFFECT OF CHEESE COMPOSITION & COOLING METHOD


===========-======================================================================================I I Budget I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I =========== 1========1========1=========1 1========1========1=========1 I========!========!========= I I Salaries/ I I I I I I I II I I II Wages I 15238 15238 0 10321 10321 0 4917 4917 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I 2493 2493 0 1689 1689 0 805 805 0 ----------- 1--------1--------1---------1 1--------1--------1---------11--------1--------1---------1 I Supplies I 13379 13379 0 9062 9062 0 4317 4317 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I 10180 10180 0 0 0 0 10180 10180 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 1451 1451 0 983 983 0 468 468 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication I 0 0 0 0 0 0 0 0 0 =========== I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I TOTALS I 42742 42742 0 22055 22055 0 20687 20687 0 ==================================================================================================I I


Weste~~ Dairy Research Ce~te~



Project Title: COMPARISION BETWEEN 40 & 640 LB BLOCKS OF UNIFORM COOLING OF 640 LB BLOCKS


=========== Budg~t


Fringe

Supplies

Equipment

Travel


I TOTALS I ============

===============================z==~===================================================l J

____ Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I I======== I======== I========= II======== I======== I========= I I======== I======== I========= II I I I II I I I I I I II I 12300 7949 4351 8763 5663 3099 3537 2286 1251 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

0 102 -102 0 73 -73 0 29 -29 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

4000 3990 10 2850 2843 7 1150 1148 3 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

0 2825 -2825 0 0 0 0 2825 -2825 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

1600 1742 -142 1140 1241 -101 460 501 -41 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 77 -77 0 55 -55 0 22 -22 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 17900 16687 1213 12752 9875 2877 5148 6812 -1664

=====================================================================================!I



Dairy Center Director: Dr. Paul A. Savello Principal Investigator: MCMAHON DONALD J

Project Title: VARIATIONS IN CASIN COMPOSITION OF MILK HIGH YIELD, LOW MOISTURE CHEESE FROM HOMOGENJ ZED UF MILK



Summary Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I =========== I======== I======== I========= II======== I======== I========= II======== I======== I========= I I Salaries/ I I I II I I II I I I I Wages I 30264 47808 -17544 23415 36989 -13574 6849 10819 -3970 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I 4842 6164 -1322 3746 4769 -1023 1096 1395 -299 ------------ 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 1 40950 22704 18246 31683 17566 14117 9267 5138 4129 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I 0 778 -778 0 0 0 0 778 -778 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 3300 504 2796 2553 390 2163 747 114 633 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication 1 0 644 -644 0 498 -498 0 146 -146 =========== I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I TOTALS I I 79356 78602 754 61398 60212 1186 17958 18389 -431 ==================================================================================================!I

• • • ANNUAL FINANCIAL REPORT BY' PROJECT

Waste~~ DaXry Rese~ch Ce~te~


Dairy Center Director: Dr. Paul A. Savello Principal Investigator: MAHONEY ARTHUR W

Project Title: EVALUATION OF IRON-PROTEIN COMPLEXES IN IRON-FORTIFIED DAIRY PRODUCTS



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========I I======== I======== I========= I I======== I======== I========= I I======== I======== I========= I I Salaries/ I I I I I I I I I I I I I I Wages 1 1 79545 78507 1037 60844 60050 794 18701 18457 244 -----------1 1-----~--1--------1---------1 1--------1--------1---------11--------1--------1---------1 I Fringe I 1 27090 5465 21625 20721 4180 16541 6369 128~ 5084 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Supplies I I 11125 13385 -2260 8510 10238 -1729 2615 3147 -531 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I I 0 0 0 0 0 0 0 0 0 -----------11--------1--------1---------11--------1--------1---------1 1--------1--------1--~------1 I Travel I I 5100 5852 -752 3901 4476 -575 1199 1376 -177 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! I 600 510 90 459 390 69 141 120 21 ===========I I -------1 ------1 -------1 I -------1 -------1 --~-----1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I I TOTALS I I 123459 103719 19741 94434 79334 15100 29025 24384 4641 =================================================================================·================1 l





Project Title: A NEW METHOD FOR MEASURING SYNERESIS OF RENNETED GELS APPLIED TO DEVELOPMENT OF CHEE~ E



Summary !Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I ===========II======== I======== I========= II======== I======== I========= II======== I======== I========= II Salaries/ I I I I I I I I I I I I I I Wages I I 3345 3345 0 2192 2192 0 1153 1153 0 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I I 95 95 0 62 62 0 33 33 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I I 1080 1080 0 708 708 0 372 372 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I 1 0 0 0 0 0 0 0 0 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I I 962 962 0 630 630 0 331 331 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! I 0 0 0 0 0 0 0 0 0 ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

II I I II I I II I I II TOTALS I I 5481 5481 0 3592 3592 0 1889 1889 0 ==================================================================================================I I


Wes~r~ Dairy Research Ce~ter



Project Title: COOLING RATE OF CHEDDAR CHEESE: COMPARISON BETWEEN 40 & 640 LB BLOCKS OF UNIFORM COC LING OF 640 LB BLOCKS


===========-======================================================================================I I Budget


Fringe

Supplies

Equipment

Travel

Publication! ===========I

I TOTALS I ============

I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I

========I========!========= I 1========1========1=========1 I======== I======== I========= I I I I II I I II I I II

20442 16182 4260 13569 10742 2828 6873 5440 1432 -~------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

1022 2614 -1592 678 1735 -1057 344 879 -535 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

12755 15834 -3079 8467 10511 -2044 4288 5324 -1035 --------1--------1---------1 1----~---1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------1 1-~------1--------1---------1 1--------1--------1---------1 I

1600 1759 -159 1062 1168 -105 538 591 -53 --------1--------1---------1 1--------l--------l---------ll--------l--------l---------l I

200 211 -11 133 140 -7 67 71 -4 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 36019 36601 -582 23909 24295 -386 12110 12305 -196

=====================================================================================!I



Dairy Center Director: Dr. Paul A. Savello Principal Investigator: MCDANIEL MINA R

Project Title: OPTIMIZATION OF THE SENSORY QUALITIES OF FLAVORED YOGURT



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========1 1========1========1=========1 1========1========1=========1 1========1========1=========1 I Salaries/ I I I I I I I I I I I I I I Wages I 20092 ~9668 424 13361 13079 282 6731 6589 142 ----------- 1--------1--------1---------11--------1--------1---------1 1--------1--------1---------1 I Fringe I 3885 6755 -2870 2584 4492 -1908 1301 2263 -961 ----------- 1--------1--------1---------1 1--------1--------1---------11--------1--------1---------1 I Supplies I 7050 4467 2583 4688 2971 1718 2362 1496 865 ----------- 1--------1--------1---------1 1--------1--------1---------11--------1--------1---------1 I Equipment I 0 0 0 0 0 0 0 0 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 1100 1237 -137 732 823 -91 369 414 -46 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication I 0 0 0 0 0 0 0 0 0 =========== I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I I 32127 32127 0 21364 21364 0 10763 10763 0 ==================================================================================================1 I





Project Title: PREDICTION & DETERMINATION OF THE EFFICACY OF NISIN IN DAIRY FOODS


Budget Summary

=~=====================c,:c=====c===c==~===========================================================l J

I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========

Salaries/ Wages

Fringe

Supplies

Equipment

Travel

Publication ===========

TOTALS ============

1========1========1=========1 1========1========1=========1 1========1========1=========1 I I I I I I I I I I I I I I

18574 27188 -8614 12354 18083 -5729 6220 9105 -2885 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

929 10133 -9204 618 6739 -6121 311 3393 -3082 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 l

34134 11283 22851 22703 7504 15198 11431 3779 7653 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------1 1--------1--------1---------J 1--------l--------1---------1 I

1500 737 763 998 490 508 502 247 256 --------1--------1---------1 1--------1--------1---------1 l--------l--------l---------1 I

0 0 0 0 0 0 0 0 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 55137 49340 5797 36672 32816 3855 18465 16524 1941

==================================•============~===•=================================1 I




Project Title: FUNCTION OF WHEY PROTEINS & LACTOSE IN AGE GELATION OF ULTRA-HIGH TEMPERATURE STERILJ ZED MILK CONCENTRATE


===========-======================================================================================!I Budget I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I =========== ========!========!=========!I======== I======== I========= I I======== I========!========= I I Salaries/ I I II I I II I I I I Wages 20400 24905 -4505 13566 16562 -2996 6834 8343 -1509 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe 6200 6125 75 4123 4073 50 2077 2052 25 ----------- --------1--------1---------11--------1--------1---------1 1--------1--------1---------1 I Supplies 19800 18009 1791 13167 11976 1191 6633 6033 600 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 0 480 -480 0 0 0 0 480 -480 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel 3600 0 3600 2394 0 2394 1206 0 1206 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! 0 481 -481 0 320 -320 0 161 -161 ===========! -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I 50000 50000 1 33250 32930 320 16750 17069 -319 ==================================================================================================!I

--------------------------------------------------




Project Title: ACID WHEY UTILIZATION: FUNCTIONAL PROPERTIES OF A FOOD GRADE STABILIZER PRODUCED BY LACTOBACILLUS PLANTARUM FROM ACID WHEY


·====~======

Budget Summary

============ Salaries/ Wages

Fringe

Supplies

Equipment

Travel


I TOTALS I ============

=~========·~~======================~============;;====================================I I ____ Tot a 1 Funds I I NDPRB Funds I I Loca 1 Funds I I

Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ========1========1=========1 1========1========1=========1 1========1========1=========1 I

I I I I I I I I I I I I 11220 14847 -3627 8497 11244 -2747 2723 3604 -880

--------1--------1---------1 1--------1-------~1---------1 1--------1--------1---------1 I 5857 3994 1863 4435 3025 1411 1422 969 452

--------1~-------1---------1 1--------1--------1---------1 1--------1--------1---------1 I 19000 22389 -3389 14389 16955 -2566 4611 5434 -822

--------1--------1---------1 1----~~--1--------1---------1 1--------1--------1---------1 I 5000 0 5000 0 0 0 5000 0 5000

--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I l8oo 2147 -347 1363 1626 -263 437 521 -84

--------1--------1---------1 1--------1--------1--------~1 1--------1--------1---------1 I 500 0 500 379 0 379 121 0 121

-------1 ------1 -------1 I -------1 -------1 -------~1 I -------1 ----~--1 --------1 I I I II I I II I I II

43377 43377 0 29063 32849 -3786 14314 10528 3786 =====================================================================================I I


Weste~~ Da±ry Rese~ch Ce~te~



Project Title: MEMBRANE FRACTIONATION OF IMMUNOGLOBULINS FROM MILK & WHEY


=========== ======================================================================================!I Budget I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent 1 Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I =========== I======== I======== I========= II======== I======== I========= II======== I======== I========= I I Salaries/ I I I I I I I II I I I I Wages 1 20800 20800 0 0 0 0 20800 20800 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I 6900 6900 0 0 0 0 6900 6900 0 ----------- 1--------l--------1---------1 1--------l--------1---------1 1--------1--------1---------1 I Supplies I 4347 4345 2 0 0 0 4347 4345 2 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I 0 0 0 0 0 0 0 0 0 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I 0 0 0 0 0 0 0 0 0 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! I 0 2 -2 0 0 0 0 2 -2 ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I I TOTALS I I 32047 32047 0 0 0 0 32047 32047 0 ==================================================================================================I I


Weste::r:n.. Da.i:ry Resea.::rc::h C::e:n..te::r



Project Title: DEVELOPMENT OF A PROCESS FOR' PRODUCTION OF UF MILK RETENTATE POWDER

Proj~ct Status: Active USU Project Number: 210 USU Account Number: 547899

=========== ==========~==================c:sc=•===================================================l I Budg~t I Total Funds I I NDPRB Funds I I Local Funds I I

Summary Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I =========== 1========1========1=•=======11====•===1========1=========1 1========1========1=========1 I Salaries/ I I I I I I I II I I I I Wages 1 70897 49577 21320 48191 33700 14492 22706 15878 6828 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Fringe I 16918 9717 7201 11500 6605 4895 5418 3112 2306 ----------- 1--------1--------1---------1 J--------1--------1---------1 1--------1--------1---------1 I Supplies I 34605 35051 -446 23522 23826 -303 11083 11226 -143 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment 1 2000 0 2000 0 0 0 2000 0 2000 ----------- 1--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel 1 1000 837 163 680 569 111 320 268 52 ----------- 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! 1 0 480 -480 0 326 -326 0 154 -154 ===========!I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

II I I II I I II I I II TOTALS I I 125420 95662 29758 83893 65025 18868 41527 30637 10890 ==================~===~===========================================================================1 I





Project Title: CONTROLLING AGE GELATION OF UHT STERILIZED MILK CONCENTRATES



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========I 1========1========1=========1 1========1========1=========1 1========1========1=========1 I sa 1 aries I I I I I I I I I I I I I I I Wages I I 35000 15717 19283 24109 10826 13283 10891 4890 6000 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I I 7140 15 7125 4918 10 4908 2222 5 2217 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Supplies I I 29000 14078 14922 19976 9698 10279 9024 4381 4643 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I I 2000 480 1520 0 0 0 2000 480 1520 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I I 0 342 -342 0 236 -236 0 106 -106 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! I 0 39 -39 0 27 -27 0 12 -12 ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I


• • • ANNUAL F~NANCIAL REPORT BY PROJECT

Westerr:L Da.i.ry Research Cer:Lter



Project Title: CAUSES & PREVENTION OF STICKY TEXTURE IN MOZZARELLA CHEESE


======~====================================•==•===================================================I I Budget I I Total Funds I I NDPRB Funds I I Local Funds I I

Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I ===========I ========I o:======•= I========= II =• .. ====:= I======== I========= II======== I======== I========= I I Salaries/ I I I I I I I II I I I I Wages I 14000 14438 -438 9380 9674 -294 4620 4765 -145 -----------1 --------1--------1---------1 1--------1-------~1---------1 1--------1--------1---------1 I Fringe I 2380 2794 -414 1595 1872 -277 785 922 -137 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I 3000 2281 719 2010 1528 482 990 753 237 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I 0 0 0 0 0 0 0 0 0 -----------1 --------1--------1---------1 1--------1--------1---------11--------1--------1---------1 I Travel I 0 0 0 0 0 0 0 0 0 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! 0 102 -102 0 68 -68 0 34 -34 ===========I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I II I I II I I II TOTALS I 19380 19615 -235 12985 13142 -157 6395 6473 -78 ============ ==========================~==========================================================I I




Dairy Center Director: Dr. Paul A. Savello Principal Investigator: SANDINE WILLIAM E

Project Title: GROWTH OF BIFIDOBACTERIA IN MILK: ASSOCIATION WITH STREPTOCOCCUS THERMOPHILUS & LAC1 OBACILLUS SPECIES AS MEASURED BY GENETIC & ENZYMATIC PROBES


==================================================================================================!I Budget


Fringe

Supplies

Equipment

Travel


I TOTALS I ============

I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I

========1========1=========1 1========1========1=========1 1========1========1=========1 I I I II I I II I I II

19935 16748 3187 13356 11221 2135 6579 5527 1052 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

6734 3807 2927 4512 2551 1961 2222 1256 966 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

22100 12617 9483 14807 8453 6354 7293 4164 3129 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

3000 303 2697 2010 203 1807 990 100 890 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 51768 33475 18293 34685 22428 12256 17083 11047 6037

=====================================================================================!I



Annual Report Ending: June 30, 1992, Final ·Project Term: 09/01/90- 06/30/92

Dairy Center Director: Dr. Paul A. Savello Principal Investigator: PENNER MICHAEL

Project Title: UTILIZATION OF ACID WHEY AS A SUBSTRATE FOR THE PRODUCTION OF FOOD GRADE CELULASES


=========== Budget


Fringe

Supplies

Equipment

Travel


I TOTALS I ============

======================================================================================I I I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance !Allocated! Spent I Balance IAllocatedl Spent I Balance I I

========1========1=========1 1========1========1=========1 I========(======== I========= I I I I I I I I II I I II

9261 8394 867 6205 5624 581 3056 2770 286 --------1--------1---------1 1--~~----1--------1---------1 1--------1--------1---------1 I

2972 4141 -1169 1991 2775 -783 981 1367 -386 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

7750 7448 302 5193 4990 203 2558 2458 100 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1~-------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

600 600 0 402 402 0 198 198 0 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 20583 20583 0 13791 13791 0 6792 6792 0

=====================================================================================!I

------------------------------------------------





Project Title: CHARACTERIZATION OF MILK PROTEOLYSIS BY LACTOCOCCAL STARTER CULTURE STRAINS USING AMJ NO ACID ANALYSIS



Summary !Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I ===========I ========I======== I========= II======== I======== I========= II======== I======== I========= II Salaries/ I I I II I I II I I I I Wages I 19650 5500 14150 13166 3685 9481 6485 1815 4670 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I 4750 0 4750 3183 0 3183 1568 0 1568 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Supplies I 15500 373 15127 10385 250 10135 5115 123 4992 -----------1 --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment I o 4343 -4343 0 0 0 0 4343 -4343 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel I 5000 0 5000 3350 0 3350 1650 0 1650 -----------1 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! 200 0 200 134 0 134 66 0 66 ===========I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I TOTALS I 45100 10215 34885 30217 3935 26282 14883 6281 8602 ============ =====================================================================================!I

• • • ANNUAL FINANCIAL REPORT BY PRc::::>JECT



Project Title: IDENTIFYING BATCH OF ORIGIN OF FINISHED CHEESE MADE IN CONTINUOUS PROCESSES



Summary !Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I =====:...::====II======== I======== I========= II====-==== I======== I========= II======== I======== I========= II Salaries/ I I I I II I I II I I II Wages I I 10000 3245 6755 6700 2174 4526 3300 1071 2229 ------·-----1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe I I 0 0 0 0 0 0 0 0 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies I I 1000 561 439 670 376 294 330 185 145 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Equipment I I 0 0 0 0 0 0 0 0 0 -----------1 1--------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Travel I I 1000 859 141 670 575 95 330 283 47 -----------11--------1--------1---------11--------1--------1---------11--------1--------1---------1 I Publication! I 0 0 0 0 0 0 0 0 0 ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I





Dairy Center Director: Dr. Paul A. Savello Principal Investigator: CORNFORTH DAREN P

Project Title: COMPARATIVE EFFECTS OF WHEY PROTEIN CONCENTRATE, LACTOSE, SALT, PHOSPHATE & PH ON COC KED YIELD, BIND & ACCEPTABILITY OF TURKEY ROLLS & BONELESS HAMS

Project Status: ACTIVE USU Project Number: 241 USU Account Number: 547950


Summary Allocated! Spent I Balance !Allocated! Spent I Balance !Allocated! Spent I Balance I I =========== ========1========1=========1 1========1========1=========1 1========1========1=========1 I Salaries/ I I II I I II I I I I Wages 17000 8383 8617 11390 5617 5773 5610 2766 2844 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe 2720 0 2720 1822 0 1822 898 0 898 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 4270 697 3573 2861 467 2394 1409 230 1179 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 0 0 0 0 0 0 0 0 0 ------------ --------1--------1---------1 1----~---1--------1---------1 1--------1--------1---------1 I Travel 0 150 -150 0 101 -101 0 50 -50 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication! 0 0 0 0 0 0 0 0 0 ===========! -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I I I I I I I I I I I I TOTALS I 23990 9230 14760 16073 6184 9889 7917 3046 4871 ============-=====================================================================================!I


Weste~~ Dairy Research Ce~te~



Project Title: THE INFLUENCE OF PREADSORBED PROTEIN ON ADHESION OF LISTERIA MONOCYTOGENES TO DAIRY I OOD CONTACT SURFACES

Project Status: ACTIVE USU Project Number: 203 USU Account Number: 547951

=========== Budget


Fringe

Supplies

Equipment -----------1 Travel I -----------1 Publication! ===========I

I TOTALS I ============

======================================================================================!I I Total Funds I I NDPRB Funds I I Local Funds I I Allocated! Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I

========1========1=========1 1========1========1=========1 1========1========1=========1 I I I II I I II I I II

29902 14400 15502 20034 9648 10386 9868 4752 5116 --------1--------1---------11--------1--------1---------1 1--------1--------1---------1 I

885 2048 -1163 593 1372 -780 292 676 -384 --------1--------1---------11--------1--------1---------1 1--------1--------1---------1 I

26966 13502 13464 18067 9046 9021 8899 4456 4443 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

1000 79 921 670 53 617 330 26 304 --------1--------1---------11--------1--------1---------11--------1--------1---------1 I

0 0 0 0 0 0 0 0 0 -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II 58753 30029 28724 39365 20120 19245 19388 9910 9479

=====================================================================================1 I


Waste~~ Dairy Research Ce~te~



Project Title: USING WHEY FOR IMPROVEMENT OF EXPOSED SUBSOILS, SODIC & SALINE-SODIC SOILS



Summary IAllocatedl Spent I Balance IAllocatedl Spent I Balance IAllocatedl Spent I Balance I I =========== ========I======== I========= I I======== I======== I========= I I======== I======== I========= II Salaries/ I I II I I II I I II Wages 15500 12893 2607 10385 8638 1747 5115 4255 860 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Fringe 0 177 -177 0 118 -118 0 58 -58 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Supplies 3500 58 3442 2345 39 2306 1155 19 1136 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Equipment 0 0 0 0 0 0 0 0 0 ----------- --------1--------1---------11--------1--------1---------11--------1--------1---------1 I Travel 2000 772 1228 1340 517 823 660 255 405 ----------- --------1--------1---------1 1--------1--------1---------1 1--------1--------1---------1 I Publication 0 0 0 0 0 0 0 0 0 =========== -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

I I II I I II I I II TOTALS I 21000 13900 7100 14070 9313 4757 6930 4587 2343 ==================================================================================================I I


W este::r-r1.. Da.i:J::"-y Resea.:r:-c:h C er1.. te::r-

S u. m ma.:r:-y Tot.a.l.s fo::r- Al.J... P::r-o .:jec::ts


Dairy Center Director: Dr. Paul A. Savello

Project Title: SUMMARY TOTALS FOR ALL WESTERN DAIRY CENTER PROJECTS

==================================================================================================1 I Budget I I Total Funds I I NDPRB Funds _____ ! I Local Funds I I

Summary !Allocated! Spent I Balance !Allocated! Spent I Balance IAllocatedl Spent I Balance I I ===========I I -------1 ------1 -------1 I -------1 -------1 --------1 I -------1 -------1 --------1 I

II I I II I I II I I II Grand Totals2,753,449 2,436,488 316,961 1,775,451 1,567,234 208,217 977,998 869,254 108,744 ============~·========•========•===•===========•==================================================I I

-------------------------------------------------------------------,

• • • Western Dairy Center Listing of Funded Projects By Account Number


Project Project Account Proj Principal Research Research Start Ending Number Num Investigator Area Title Date Date FY88 FY89 FY90 FY91 FY92 FY93 FY94 FY95 FY96

547766 189 Hansen, C.L. Ultrafiltration Cogeneration of Biogas and 07/01187 06/30/89 29589 29589 0 0 0 0 0 0 0 and Reverse Single Cell protein From Osmosis Ultrafiltration Permeate

and Whey

547767 188 Broadbent, J. Microbiology Cloning the Nisin and Other 07/01/87 06/30/93 41680 34440 32105 33711 35396 0 0 0 0



547768 182 Mahoney, A.W. Product Quality Iron fortification of cheese 08/01187 06/30/90 20944 21753 0 0 0 0 0 0 0 curd

547769 190 Richarctson, G. Product Quality Acquisiton of Zymark II 07/01187 06/30/90 5000 0 0 0 0 0 0 0 0 Robot for Laboratory Automation Studies

547780 181 Richardson, G. Curd Improving Yield and 07/01187 06/30/91 22620 22620 22620 0 0 0 0 0 0


547781 183 Emstrom, C.A. Ultrafiltration Continuous Production of 09/01187 08/31189 8700 0 0 0 0 0 0 0 0 and Reverse Cottage Cheese From Osmosis Ultra-filtrated skim milk

Retenate

547782 184 Cornforth, D.P. Product Quality Evaluation of Milk Proteins 07/01187 6/30/90 15327 16512 0 0 0 0 0 0 0 as Whitening Agents in Processed Meats and Poultry Products

547783 186 Olsen, R.L. Curd Interaction of Protein and 07/01/87 06/30/89 21750 21410 0 0 0 0 0 0 0 Formation/ Polysaccharides in Chymosin Cheese Tech. and Acid Coagulation of Milk



547784 187 Richardson, G. Curd Improved Control of Cheese 07/01187 04/30/92 35300 14000 14000 0 0 0 0 0 0

Formation/ Manufacture Through Vat Cheese Tech. Monitoring

547785 191 Savello, P.A. Administrative Western Dairy Foods 07/01187 06/30/92 20000 30000 50000 50000 50000 50000 50000 50000 50000

Expenses Research Center Administrative Account

547802 192 Ernstrom, C.A. Curd Effect of Milk Clotting 07/01187 06/30/88 37660 0 0 0 0 0 0 0 0

Formation/ Enzymes on the Curing and Cheese Tech. Quality of Cheddar Cheese

547804 188 Sandine, W.E. Microbiology Cloning the Nisin and Other 07/01187 12/31193 44565 43033 45274 47537 49914 0 0 0 0



547810 194 Sandine, W.E. Microbiology Characterization of 07/01187 06/30/90 18110 18573 18573 0 0 0 0 0 0

of Starter Bacteriophage Receptor Cultures Sites of Lactic

Streptococci

547811 195 Bodyfelt, F.W. Product Quality Rapid Assay for Heat 08/01/87 12/31/90 18285 3035 15260 0 0 0 0 0 0

Resistant Microbial Proteases in Raw Milk by a Simple Casein Denaturation Method

547812 196 Bodyfelt, F.W. Microbiology Production of Omega-3 Fatty 09/01187 12/31190 24350 5415 24890 0* 0 0 0 0 0

of Starter Acids by Genetically Cultures Altered Fungi and Lactic

Acid Bacteria

547813 197 McGuire, J. Product Quality Characterization of the 01/01/88 12/31/90 20910 31043 24643 0 0 0 0 0 0

Post-Absorbtive Behavior of B-Lactoglobulin For Control of Spore and Microbial Adhesion



547814 198 Sandine, W.E. Microbiology Studies on the Growth and 07/01/87 06/30/89 18110 18573 0 0 0 0 0 0 0 of Starter Survival of Bifidobacterium Cultures Species in Milk

547822 183 Ogden, L.V. Ultrafiltration Continuous Production of 09/01187 08/31189 0 8200 0 0 0 0 0 0 0 and Reverse Cottage Cheese From Osmosis Ultrafiltrated Skim Milk

Retenate

547823 199 Ogden, L.V. Product Quality Method for Identifying 07/01188 06/30/90 0 6100 6100 0 0 0 0 0 0 Batch of Origin of Semi-continuous Cheese Processes

547825 201 Brown, R.J. Product Quality Application of Fourier 07/01188 12/31191 0 73350 25205 26366 0 0 0 0 0


547827 267 Brown, R.J. Product Quality Estimation of Individual 07/01188 12/31193 0 145425 50594 51874 0 0 0 0 0


547828 204 Savello, P.A. Ultrafiltration Properties of Low-fat Yogurt 08/01188 09/30/91 0 28500 29700 0 0 0 0 0 0

and Reverse Manufactured from Ultra-Osmosis filtered and Ultra-high Temp-

Treated Milk




547834 202 Torres, J.A. Curd Cheddar Cheese Blocks: 07/01/88 12/31190 0 26530 16212 0 0 0 0 0 0




547838 206 Hansen, C.L. Curd Comparison Between 40 and 11/01/88 12/31/91 0 2300 11000 4600 0 0 0 0 0 Formation/ 640 lb Blocks of Uniform Cheese Tech. Cooling of 640 lb Blocks

547839 200 McMahon, D.J. Ultrafiltration Variations in Casein 09/01/88 06/30/92 0 24936 26543 27877 0 0 0 0 0 and Reverse Composition of Milk High Osmosis Yield, Low Moisture Cheese

From Homogenized UF Milk

547841 208 Mahoney, A.W. Product Quality Evaluation of Iron-Protein 10/04/88 12/31/92 0 35505 /80901138442//41422/ 0 0 0 0 Complexes in Iron-Fortified Dairy Products

547848 210 Hansen, C.L. Curd A New Method for Measuring 03/01/89 03/01/91 0 23912 0 0 0 0 0 0 0 Formation/ Syneresis of Renneted Gels Cheese Tech. Applied to Development of

Cheese

547862 206 Torres, J.A. Curd Cooling Rate of Cheddar 11/01/88 12/31/91 0 11549 18320 2475 0 0 0 0 0 Formation/ Cheese: Comparison Between Cheese Tech. 40 and 640 lb Blocks of



547864 207 Daeschel, M.A. Microbiology Prediction and 07/01/88 mn.wn 0 18230 21177 0 /8240/ 0 0 0 0 of Starter Determination of the Cultures Efficacy of Nisin in Dairy

Foods

547867 211 McMahon, D.J. UHT Function of Whey Proteins 07/01/89 06/30/92 0 0 25000 25000 0 0 0 0 0 Processing and Lactose in Age Gelation

of Ultra-High Temperature Sterilized Milk Concentrate



547877 212 Torres, J.A. Product Quality Acid Whey Utilization: 07/01189 06/30/91 0 0 l2o58oll22797l 0 0 0 0 0 Functional Properties of a Food Grade Stabilizer Produced by Lactobacillus Plantarum from Acid Whey

547898 204 Savello, P.A. UF/RO Membrane fractionation of 07/01190 06/30/91 0 0 0 33500 0 0 0 0 0 immunoglobulins from milk and whey

547899 210 Hansen, C.L. UF/RO Development of a process 07/01190 06/30/93 0 0 0 61405 64415 0 0 0 0 for production of UF milk retentate powder

547900 211 McMahon, D.J. UHf Controlling age gelation of 07/01190 06/30/93 0 0 0 34440 38740 0 0 0 0 Processing UHT sterilized milk

concentrates

547901 181 Richardson, G. Product Quality Causes and prevention of 07/01190 06/30/91 0 0 0 19380 0 0 0 0 0 sticky texture in Mozzarella cheese

547902 198 Sandine, W.E. Microbiology Growth ofbifidobacteria in 07/01190 12/31192 0 0 0 1254531 26315 0 0 0 0 of Starter milk: Association with Cultures Streptococcus thermophilus

and Lactobacillus species as measured by genetic and enzymatic probes

547903 212 Penner, M. Product Quality Utilization of acid whey as 07/01190 06/30/92 0 0 0 20583 0 0 0 0 0 a substrate for the production of food grade cellulases

547910 243 Brown, R.J. Microbiology Characterization of milk 07/01190 12/31193 0 0 0 22000 23100 0 0 0 0 of Starter proteolysis by lactococcal Cultures starter culture strains using

amino acid analysis



547951 203 Daeschel, M. Microbiology Influence of Preadsorbed 07/01/91 06/30/94 0 0 0 0 30354 28399* 29544 0 0 of Starter Protein on Adhesion of Cultures Listeria monocytogenes

to Dairy Food Contact Surfaces

547936 183 Ogden, L.V. Product Quality Identifying Batch of Origin 07/01/91 6/30/93 0 0 0 0 12000 0 0 0 0 of Finished Cheese Made in Continuous Processes

547950 241 Cornforth, D.P. Product Quality Comparative Effects of Whey 07/01/91 12/31/92 0 0 0 0 23990 0 0 0 0 Protein Concentrate (WPC), Lactose, Salt, Phosphate, and pH on Cooked Yield, Bind, and Acceptability of Turkey Rolls and Boneless Hams

547952 263 Hansen, C.L. Whey Utilization Using Whey for Improve- 07/01191 06/30/93 0 0 0 0 21000 14000 0 0 0 ment of Exposed Subsoils and Sodic and Saline-Sodic Soils

547958 199 Schelling, G.R. Directed Using a Natural Process to 07/01/92 06/30/94 0 0 0 0 0 19838 19438 0 0 Product Quality Improve Milk Quality and Extend Milk Shelf-Life Through the Reduction in Lipid Oxidation and Off-Flavors with Tocopherol (Vitamin E) Supplementation to Dairy Cows

547961 211 McMahon, D.J. Fundamental Function of Whey Proteins 07/01/92 06/30/94 Thermal Process and Lactose in Age Gelation of

0 0 0 0 0 22075 26000 0 0

UHT -Processed Milk Concen-trate - Part 2

547962 205 McMahon, D.J. Practical Effects of Iron Fortification 07/01/92 06/30/93 0 0 0 0 0 26700 0 0 0 Product Quality on Chemical, Physical, Microbiological Nutrition and Nutritional Properties of Yogurt

• Account Proj Number Num

Principal Investigator

547963 244 Weimer, B.

547964 244 Weimer, B.

547965 204 Geller, B.

547970 263 Hansen, C.L.

Research Area

Fundamental Starter Cultures

Practical Starter Cultures

Fundamental Starter Cultures

Practical Milk Proteins

Research Title

Project Start Date

• Project Ending Date

Interactions Between Milk 07/01/92 06/3 0/94 0 Proteins, Starter Cultures, and Hydrocolloidal Milk Fat Replacers

Production of Extracellular 07/01192 06/30/94 0 Proteases of Brevibacterium linens for Use in Lowfat Cheese

Bacteriophage-Resistance Gene Replacement in Lactococcus lactis

Extrusion Processing of Milk Proteins

07/01192 06/30/95 0

07/01/92 06/30/94 0

547978 200 McMahon, D.J. Practical Rheology and Microstructure 07/01192 06/30/94 0

547980 202 Hansen, C.

Brown, R.J.

Brown, R.J.

Cheese Techno!. of Mozzarella Cheese

Practical Development of High Protein04/0l/93 03/31/94 0 Cheese Techno!. Low-fat Fermented Foods

from Yogurt Cheese

Fundamental Whey Proteins

Fundamental Milk Proteins

Purification of Monospecific 06/01193 05/31/95 0 Polyclonal Antibodies from Bovine Cheese Whey

Milk Protein Interactions and 07/01193 06/30/96 0 Gelation During Thermal Processing

0

0

0

0

0

0

0

0

• Project Funding Allocated Thru FY 1996

0 0 0 42620 49276 0 0

0 0 0 35625 39293 0 0

0 0 0 40860 41571 44228 0

0 0 0 47200 29650 0 0

0 0 0 24600 43440 0 0

0 0 0 0 39242 0 0

0 0 0 44875 42360 0 0

0 0 0 0 60735 67925 68026

***TOTAL*** 402900 762722 492722l56774oll424886l396792 470549 162153 118026

LEGEND:


* - Decrease in project funding approved for FY s

_( __ ) - Extension (no cost) approved

L ..... ) - Project revision approved

( __ )- Projected funding

•

•

•

WESTERN DAIRY FOODS

RESEARCH CENTER

ANNUAL MEETING Oregon State University, Corvallis

July 9-1 0, 1992

•

•

•

TABLE OF CONTENTS

Meeting Agenda ............................................................................................................ 1

Operational Advisory Committee ............................................................................... 5

Principallnvestigators .................................................................................................. 7

WDFRC Budget Activity ............................................................................................... 8

Listing of Projects by Account Number ..................................................................... 9

Annual Project Reports by Research Area:

Product Quality and Nutrition

9:00a.m.

9:15a.m.

9:30a.m.

9:45a.m.

10:00 a.m.

10:15 a.m.

10:30 a.m .

Identifying batch of origin of finished cheese

made in continuous processes ............................................ 15


Utilization of acid whey as a substrate for the

production of food grade cellulases .................................... 1 7


Speaker: Soren Nordmark (M.S. program/OSU)

Comparative effects of whey protein concentrate

(WPC), lactose, salt, phosphate, and pH on cooked

yield, bind, and acceptability of turkey rolls and

boneless hams ........................................................................ 18


Application of Fourier transform infrared technology

to milk and dairy products ..................................................... 19


Evaluation of iron-protein complexes in iron-fortified

dairy products .......................................................................... 22


Speaker: Mohan Reddy (Post doctorai/USU)

Estimation of individual milk proteins and genetic

variants by multicomponent analysis of amino

acid profiles .............................................................................. 25


BREAK

Curd Formation/Cheese Technology • 10:45 a.m. Improved control of cheese manufacture through vat

monitoring ................................................................................ 27


11:00 a.m. Comparison between 40 and 640 lb blocks of uniform

cooling of 640 lb blocks ......................................................... 29


Ultra-high Temperature Processing 11:15 a.m. Function of whey proteins and lactose in age gelation of

ultra-high temperature sterilized milk concentrate .......... 34


11:30 a.m. Controlling age gelation of UHT sterilized milk

concentrates ............................................................................ 37


Whey Utilization 11 :45 a.m. Using whey for improvement of exposed subsoils

• and sodic and saline-sodic soils ......................................... 39

•


NOON LUNCH

Microbiology of Starter Cultures 1:30 p.m. Cloning the nisin and other genes of lactic streptococci

into Leuconostoc species and amplification of nisin

1:45 p.m.

production ................................................................................ 41


Purification of a bacteriocin from Pediococcus

pentosaceus and genetic transfer of plasmid-borne

determinants ............................................................................ 4 7


Speaker: Xintian Ming (Ph.D. program/OSU)

ii

•

•

•

2:00p.m . Growth of Bifidobacteria in milk: Association with

Streptococcus thermophilus and Lactobacillus

species as measured by genetic and

enzymatic probes .................................................................... 51


Microbiology of Starter Cultures (continued) 2:15 p.m. Influence of preadsorbed protein on adhesion of Listeria

2:30p.m.

2:45p.m.

UF/RO 3:00p.m.

3:15p.m.

3:30p.m.

3:45p.m .

monocytogenes to dairy food contact surfaces ................. 53


Speaker: Joseph McGuire (OSU)

Characterization of milk proteolysis by lactococcal

starter culture strains using amino acid analysis .............. 55


Prediction and determination of the efficacy of

nisin in dairy foods .................................................................. 56


Speaker: Dong-Sun Jung (Ph.D. program/OSU)

Development of a process for production of UF milk

retentate powder ..................................................................... 60


High yield, low moisture cheese from homogenized

UF milk ...................................................................................... 62


Speaker: Brian Orme (Ph.D. program/USU)

Properties of low-fat yogurt manufactured from

ultrafiltered and ultra-high temperature treated milk ....... 64


Final Comments- Paul Savello

iii

•

•

•

New WCDPRT Projects, 1992·93 ..................................................................... 68

1.

2.

3.

4.

5.

6.

7.

8.

Production of Extracellular Proteases of Brevibacterium linens for Use in Lowfat Cheese ................................................................................ 69

Bacteriophage-Resistance Gene Replacement in Lactococcus lactis .................................................................................................................... 73

Purification of Monospecific, Polyclonal Antibodies from Bovine Cheese Whey .................................................................................................... 76

Rheology and Microstructure of Mozzarella Cheese ....................................... 79

Function of Whey Proteins and Lactose in Age Gelation of UHT -Processed Milk Concentrate: Part 2 ................................................... 82

Extrusion Processing of Whey Proteins (Pending) ........................................... 84

Effects of Iron Fortification on Chemical, Physical, Microbiological and Nutritional properties of Yogurt .................................. 90

Effects of pH, Salts, and Thiolation on Thermal Gelation of Caseins Alone and in Mixed Protein-Protein and Protein-Polysaccharide Gels ........................................................................................ 95 ~

Interactions Between Milk Proteins, Starter Cultures, and Hydrocolloidal Milk Fat Replacers ................................................................. 98 ~

Interaction Between Milk Proteins During Thermal Processing and Its Effects on the Physicochemical Properties of Dairy Foods .................................................................................................................. 1 03

Summary Recommendations for 1992-93 Project ~J)J)r()\1Ell ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••.••.••••...••• 1 08

iv

•

•

•


1992 ANNUAL MEETING

LASELLS STUART CENTER, OREGON STATE UNIVERSITY

MEETING AGENDA

THURSDAY, July 9, 1992

8:30a.m. Opening remarks and introductions

Paul A. Savello, Director

Floyd Bodyfelt, Liaison

9:00 to 10:30 a.m. Oral progress reports

Product Quality and Nutrition 9:00a.m.

9:15a.m .

9:30a.m.

9:45a.m.

10:00 a.m.

10:15 a.m.

Identifying batch of origin of finished cheese made in continuous processes.


Utilization of acid whey as a substrate for the production of food grade cellulases.


Speaker: Soren Nordmark (M.S. program/OSU)

Comparative effects of whey protein concentrate (WPC),

lactose, salt, phosphate, and pH on cooked yield, bind, and

acceptability of turkey rolls and boneless hams.


Application of Fourier transform infrared technology to milk and dairy products.


Evaluation of iron-protein complexes in iron-fortified dairy products.


Speaker: Mohan Reddy

Estimation of individual milk proteins and genetic variants

by multicomponent analysis of amino acid profiles. PI: Rodney Brown (USU)

1

10:30 a.m. BREAK • Curd Formation/Cheese Technology

10:45 a.m.

11:00 a.m.

Improved control of cheese manufacture through vat

monitoring.


Comparison between 40 and 640 lb blocks of uniform

cooling of 640 lb blocks.


Ultra-high Temperature Processing 11:15 a.m.

11:30 a.m.

Whey Utilization

Function of whey proteins and lactose in age gelation of

ultra-high temperature sterilized milk concentrate.


Controlling age gelation of UHT sterilized milk

concentrates.


• 11 :45 a.m. Using whey for improvement of exposed subsoils and sodic

and saline-sadie soils.

•


NOON LUNCH

Microbiology of Starter Cultures

1:30 p.m. Cloning the nisin and other genes of lactic streptococci into

Leuconostoc species and amplification of nisin production.


1:45 p.m.

2:00p.m.

Purification of a bacteriocin from Pediococcus pentosaceus

and genetic transfer of plasmid-borne determinants.


Speaker: Xintian Ming (Ph.D. program/OSU)

Growth of Bifidobacteria in milk: Association with

Streptococcus thermophilus and Lactobacillus species as

measured by genetic and enzymatic probes .


2

---------------------------------------------

Microbiology of Starter Cultures (continued)

• 2:15p.m. Influence of preadsorbed protein on adhesion of Listeria

monocytogenes to dairy food contact surfaces.


Speaker: Joseph McGuire (OSU)

2:30p.m. Characterization of milk proteolysis by lactococcal starter

culture strains using amino acid analysis.


2:45p.m. Prediction and determination of the efficacy of nisin in dairy

foods.


Speaker: Dong-Sun Jung (Ph.D. program/OSU)

UF/RO 3:00p.m. Development of a process for production of UF milk

retentate powder.


3:15p.m . High yield, low moisture cheese from homogenized UF

• milk.


3:30p.m. Properties of low-fat yogurt manufactured from ultrafiltered

and ultra-high temperature treated milk.


3:45p.m. Final Comments - Paul Savello

4:00 p.m. ADJOURN FOR SALMON BAKE!!!

5:00 - 7:00 p.m. SALMON BAKE!!!!!!

• 3

7:00 p.m . •

•

•

Operational Advisory Committee Business Meeting

1 • Acceptance of 1991 OAC Meeting Minutes

2. Barbara Blakistone (NDPRB) Presentation

3. WCDPRT Financial Report

4. NDPRB Competitive Research Proposals

Approved

5. New (1992 to 1993) WCDPRT Research

Proposals 6. WCDPRT Structure/Research - 1993 to 1996

• Response to CMAB

• WCDPRT "Program Research" and Funding

• WCDPRT "Mini-Competitive Research" and Funding

7. Set Date for WCDPRT 1994 OAC Meeting (USU)

8. Other Business 9. Adjournment

4

•

•

•


OPERATIONAL ADVISORY COMMITTEE

Pursuant to the WDFRC proposal and contract with the National Dairy Promotion and Research Board, the voting members of the Operational Advisory Committee are:

Barbara Blakistone National Dairy Promotion and

Research Board 2111 Wilson Blvd., Suite 600 Arlington VA 22201 (703) 528-4800

Rodney J. Brown, Dean College of Agriculture Utah State University Logan UT 84322-4800 (801) 750-2215

Daniel F. Farkas, Head Department of Food Science

and Technology Wiegand Hall 240 Oregon State University Corvallis OR 97331 (503) 737-3131

Jeff Strandholm Kraft, Inc. Research and Development Div. 801 Waukegan Road Glenview IL 60025 (312) 998-3717

Raj G. Narasimmon Schreiber Foods, Inc. P.O. Box 19010 Green Bay Wl54305 (414) 437-7601

5

Sheldon Pratt Oregon Dairy Products Commission 10505 S.W. Barbur Blvd. Portland OR 97219 (503) 229-5033

Paul A. Savello, Director Western Dairy Foods Res. Center Department Nutr. & Food Sciences Utah State University Logan UT 84322-8700 (801) 750-2106

Harry Papageorge 1616 Farr West Drive Ogden UT (801) 782-9383

Douglas Willrett Marschall Products P.O. Box 592 Madison WI 53701 (608) 276-3600

Boyd Gardner Utah Dairy Commission 8460 North Highway 69 Honeyville UT 84314 (801) 279-8320

•

•

•


OPERATIONAL ADVISORY COMMITTEE (continued)

Gale Moser United Dairymen of Idaho 1864 South Hulls Crossing Preston ID 83263 (208) 852-0560

Tom Holzinger Borden, Inc. 960 Kingsmill Parkway Columbus OH 43229 (614) 431-6623

6

• WESTERN DAIRY FOODS RESEARCH CENTER

PRINCIPAL INVESTIGATORS

Floyd W. Bodyfelt Mina R. McDaniel Dept. of Food Science & Technology Dept. of Food Science & Technology Oregon State University Oregon State University Corvallis OR 97331 Corvallis OR 97331

Rodney J. Brown Joseph McGuire Dept. of Nutrition & Food Sciences Dept. of Food Science & Technology Utah State University Oregon State University Logan UT 84322-8700 Corvallis OR 97331

Daren P. Cornforth Donald J. McMahon Dept. of Nutrition & Food Sciences Dept. of Nutrition & Food Sciences Utah State University Utah State University Logan UT 84322-8700 Logan UT 84322-8700

Mark A. Daeschel Lynn V. Ogden Dept. of Food Science & Technology Dept. of Food Science & Nutrition Oregon State University Brigham Young University Corvallis OR 97331 Provo UT 84602

• Bruce L. Geller Michael H. Penner Dept. of Microbiology Dept. of Food Science & Technology Oregon State University Oregon State University Corvallis OR 97331 Corvallis OR 97331

Conly L. Hansen Gary H. Richardson Dept. of Nutrition & Food Sciences Dept. of Nutrition & Food Sciences Utah State University Utah State University Logan UT 84322-8700 Logan UT 84322-8700

Jeff Broadbent William E. Sandine Dept. of Nutrition & Food Sciences Dept. of Microbiology Utah State University Oregon State University Logan UT 84322-8700 Corvallis OR 97331

Arthur W. Mahoney Paul A. Savello Dept. of Nutrition & Food Sciences Dept. of Nutrition & Food Sciences Utah State University Utah State University Logan UT 84322-8700 Logan UT 84322-8700

J. Antonio Torres Janine E. Trempy Dept. of Food Science & Technology Dept. of Microbiology Oregon State University Oregon State University • Corvallis OR 97331 Corvallis OR 97331

7

•

•

•

WDFRC BUDGET ACTIVITY 1991-1992 FISCAL YEAR

NATIONAL DAIRY PROMOTION AND RESEARCH BOARD

REGIONAUINDUSTRY SUPPORT: Utah Dairy Commission United Dairymen of Idaho Oregon Dairy Products Commission Western Dairy Farmers' Promotion Association

Kraft General Foods, Inc. Schreiber Foods, Inc. Marschai-Rhone Poulenc, Inc. Borden, Inc.

State of Utah Center of Excellence "Center for Dairy Foods Technology"

TOTAL REGIONAUINDUSTRY SUPPORT

50,000 50,000 40,000 10,000

5,000 5,000 5,000 5,000

130.000

300,000

FY92 TOTAL DAIRY RESEARCH CONTRIBUTIONS


TOTAL AVAILABLE FUNDS FOR FY91 RESEARCH

FY92 COMMITTED RESEARCH FUNDS Western Dairy Foods Research Center State of Utah Center of Excellence Administrative

(424,886) (130,000)

(50.000)

400,000

$700,000

$41,416

$741,416

TOTAL FY92 COMMITTED RESEARCH FUNDS $604,886

FY93 BALANCE FORWARD $136,530

8

• • • Western Dairy Center Listing of Funded Projects By Account Number


Project Project Accmmt Proj Principal Research Research Start Ending ~ Numb Investigator Area Title Date Date .Enj ETI.2 EY.2Q EY.2l EY.2Z .EX2l FY94 FY95 .f)'M

547766 189 Hansen, C.L. Ultrafiltration Cogeneration of Biogas and 07/01/87 06/30/89 29589 29589 0 0 0 0 0 0 0 and Reverse Single Cell protein From Osmosis Ultrafiltration Permeate

and Whey

547767 188 Broadbent, J. Microbiology Cloning the Nisin and Other 07/01/87 06/30/93 41680 34440 32105 33711 35396



547768 182 Mahoney, A.W. Product Quality Iron fortification of cheese 08/01/87 06/30/90 20944 21753 0 0 0 0 0 0 0 curd

547769 190 Richardson, G.H. Product Quality Acquisiton of Zymark IT 07/01/87 06/30/90 5000 0 0 0 0 0 0 0 0 Robot for Laboratory Automation Studies

547780 181 Richardson, G.H. Oml Improving Yield and 07/01/87 06/30/91 22620 22620 22620 0 0 0 0 0 0


547781 183 Ernstrom, C.A. Ultrafiltration Continuous Production of 09/01/87 08/31/89 8700 0 0 0 0 0 0 0 0 and Reverse Cottage Cheese From Osmosis Ultra-filtrated skim milk

Retenate

547782 184 Cornforth, D.P. Product Quality Evaluation of Milk Proteins 07/01/87 6/30/90 15327 16512 0 0 0 0 0 0 0 as Whitening Agents in Processed Meats and Poultry Products

547783 186 Olsen, R.L. Oml Interaction of Protein and 07/01/87 06/30/89 21750 21410 0 0 0 0 0 0 0 Formation/ Polysaccharides in Chymosin Cheese Tech. and Acid Coagulation of Milk

9


Project Project Account Proj Principal Research Research Start Ending Number Numb Investigator Area Title Date Date FY88 fTI2 FY90 fY.2l EY..21 ~ FY94 FY95 .EY22

547784 187 Richardson, G .H. Cutd Improved Control of Cheese 07/01/87 04/30/92 35300 14000 14000 0 0 0 0 0 0 Formation/ Manufacture Through Vat Cheese Tech. Monitoring

547785 191 Savello, P.A. Administrative Western Dairy Foods 07/01/87 06/30/92 20000 30000 30000 50000 50000 50000 50000 50000 50000 Expenses Research Center

Administrative Account

547802 192 Emstrom, C.A. Cutd Effect of Milk Clotting 07/01/87 06/30/88 37660 0 0 0 0 0 0 0 0 Formation/ Enzymes on the Curing and Cheese Tech. Quality of Cheddar Cheese

547804 188 Sandine, W.E. Microbiology Cloning the Nisin and Other 07/01/87 06/30/93 44565 43033 45274 47537 49914 0 0 0 0 of Starter Genes of Lactic Cultures Streptococci into


547810 194 Sandine, W.E. Microbiology Characterization of 07/01/87 06/30/90 18110 18573 18573 0 0 0 0 0 0 of Starter Bacteriophage Receptor Culwres Sites of Lactic

Streptococci

547811 195 Bodyfelt, F.W. Product Quality Rapid Assay for Heat 08/01/87 12/31/90 18285 3035 15260 0 0 0 0 0 0 Resistant Microbial Proteases in Raw Milk by a Simple Casein Denaturation Method

547812 196 Bodyfelt, F.W. Microbiology Production of Omega-3 Fatty 09/01/87 12/31/90 24350 5415 24890 o• 0 0 0 0 0 of Starter Acids by Genetically Culwres Altered Fungi and Lactic

Acid Bacteria

547813 197 McGuire, J. Product Quality Characterization of the 01/01/88 12/31/90 20910 31043 24643 0 0 0 0 0 0 Post-Absorbtive Behavior of B-Lactoglobulin For Control of Spore and Microbial Adhesion

10


Project Project Account Proj Principal Research Research Start Ending ~ NYmb Investigator ~ Title Date Date FY88 FY89 EY.2Q EY.2.l EY.21 .EYll FY94 ~ FY96

547814 198 Sandine, W.E. Microbiology Studies on the Growth and 07/01/87 06/30/89 18110 18573 0 0 0 0 0 0 0

of Starter Survival of Bifidobacterium Cultures Species in Milk

547822 183 Ogden, L.V. Ultrafiltration Continuous Production of 09/01/87 08/31/89 0 8200 0 0 0 0 0 0 0

and Reverse Cottage Cheese From Osmosis Ultrafiltrated Skim Milk

Retenate

547823 199 Ogden, L.V. Product Quality Method for Identifying 07/01/88 06/30/90 0 6100 6100 0 0 0 0 0 0

Batch of Origin of Semi-continuous Cheese Processes

547825 201 Brown. R.I. Product Quality Application of Fourier 07/01/88 12/31/91 0 73350 25205 26366 0 0 0 0 0


547827 267 Brown, R.J. Product Quality Estimation of Individual 07/01/88 12/31/92 0 145425 50594 51874 0 0 0 0 0


547828 204 Savello, P.A. Ultrafiltration Properties of Low-fat Yogurt 08/01/88 09/30/91 0 28500 29700 0 0 0 0 0 0

and Reverse Manufactured from Ultra-Osmosis flltered and Ultra-high Temp-

Treated Milk




547834 202 Torres, J.A. Curd Cheddar Cheese Blocks: 07/01/88 12/31/90 0 26530 16212 0 0 0 0 0 0


11


Project Project Account Proj Principal Research Research Start Ending ~ Nl!mJ2 Investigator .Arg Title Date Date FY88 m2 EY..2Q .EY..2.l FY92 FY93 FY94 FY95 ~

547838 206 Hansen, C.L. Cutd Comparison Between 40 and 11/01/88 12/31/91 0 2300 11000 4600 0 0 0 0 0 Formation/ 640 lb Blocks of Uniform Cheese Tech. Cooling of 640 lb Blocks

547839 200 McMahon, D.J. Ultrafiltration Variations in Casein 09/01/88 06/30/92 0 24936 26543 27877 0 0 0 0 0 and Reverse Composition of Milk High Osmosis Yield, Low Moistme Cheese

From Homogenized Milk

547841 208 Mahoney, A.W. Product Quality Evaluation of Iron-Protein 10/04/88 12/31/92 0 35505 l8o9oll38442l~ 0 0 0 0 Complexes in Iron-Fortified Dairy Products

547848 210 Hansen, C.L. Cutd A New Method for Measuring 03/01/89 03/01/91 0 23912 0 0 0 0 0 0 0 Formation/ Syneresis of Renneted Gels Cheese Tech. Applied to Development of

Cheese

547862 206 Torres, J.A. Cutd Cooling Rate of Cheddar 11/01/88 12/31/91 0 11549 18320 2475 0 0 0 0 0 Formation/ Cheese: Comparison Between Cheese Tech. 40 and 640 lb Blocks of



547864 207 Daeschel, M.A. Microbiology Prediction and 07/01/88 Q~.O..U9.Z 0 18230 21177 0 182401 0 0 0 0 of Starter Determination of the Cultures EffiCacy of Nisin in Dairy

Foods

547867 211 McMahon, D.J. UHf Function of Whey Proteins 07/01/89 06/30/92 0 0 25000 25000 0 0 0 0 0 Processing and Lactose in Age Gelation

of Ultra-High Temperatme Sterilized Milk Concentrate

12


Project Project Account Proj Principal Research Research Start Ending Number Numb Investigator ~ ~ Date ~ FY88 FY89 FY90 .fX.2l ~ ~ .EX24 FY95 FY96

547877 212 Torres, J.A. Product Quality Acid Whey Utilization: 07/01/89 06/30/91 0 0 l2o58oll22797l 0 0 0 0 0 Functional Properties of a Food Grade Stabilizer Produced by Lactobacillus Plantarum from Acid Whey

547898 204 Savello, P.A. UF/RO Membrane fractionation of 07/01/90 06/30/91 0 immunoglobulins from milk

0 0 33500 0 0 0 0 0

and whey

547899 210 Hansen, C.L. UF/RO Development of a process 07/01/90 06/30/93 0 0 0 61405 64415 0 0 0 0 for production of UF milk retentate powder

547900 211 McMahon, D.J. UHf Controlling age gelation of 07/01/90 06/30/93 0 0 0 34440 38740 0 0 0 0 Processing UHT sterilized milk

concentrates

547901 181 Richardson, G.H. Product Quality Causes and prevention of 07/01/90 06/30/91 0 sticky texture in Mozzarella

0 0 19380 0 0 0 0 0

cheese

547902 198 Sandine, W.E. Microbiology Growth ofbifidobacteria in 07/01/90 12/31/92 0 0 0 1254531 26315 0 0 0 0 of Starter milk: Association with Cultures Streptococcus thermophilus

and Lactobacillus species as measured by genetic and enzymatic probes

547903 212 Penner, M. Product Quality Utilization of acid whey as 07/01/90 06/30/92 0 0 0 20583 0 0 0 0 0 a substrate for the production of food grade cellulases

547910 243 Brown, R.J. Microbiology Characterization of milk 07/01/90 12/31/92 0 0 0 22000 23100 0 0 0 0 of Starter proteolysis by lactococcal Cultures starter culture strains using

amino acid analysis

13

• Account Proj Principal Number Numb Investigator

54 7933 203 Daeschel, M.

547936 183 Ogden, L.V.

Research Area

Microbiology of Starter Cultures

Product Quality

Research Title

Influence of Preadsorbed Protein on Adhesion of Listeria monocytogenes to Dairy Food Contact Surfaces

Identifying Batch of Origin of Finished Cheese Made in Continuous Processes

• Project Project Start Ending Date ~ FY88

07/01/91 06/30/94 0

07/01/91 12/30/92 0

547950 241 Cornforth, D.P. Product Quality Comparative Effects of Whey 07/01/91 12/31/92 0 Protein Concentrate (WPC), Lactose, Salt, Phosphate, and pH on Cooked Yield, Bind, and Acceptability of Turkey Rolls and Boneless Hams

547952 263 Hansen, C.L. Whey Utilization Using Whey for Improve- 07/01/91 06/30/93 0 ment of Exposed Subsoils and Sodic and Saline-Sodic Soils


FY89 fY.2Q .EY.2.l ~ EY.2l FY94

0 0 0 30354 28399* 29544

0 0 0 12000 0 0

0 0 0 23990 0 0

0 0 0 21000 14000 0

***TOTAL*** 4022oo 762722 492722 ls6n4oll424ss61 93671 79544

LEGEND:


* - Decrease in project funding approved for FY s

L___l - Extension (no cost) approved

L .... J- Project revision approved

L_)- Projected funding

14

• ~ FY96

0 0

0 0

0 0

0 0

•

•

•

NEW WCDPRT PROJECTS, 1992-93

Eight (8) new projects are recommended for WCDPRT funding beginning

July 1, 1992. These projects were reviewed by the Technical Advisory

Committee (TAC) of the WCDPRT during March to June, 1992. Comments from

TAC members were compiled and submitted to the Principal Investigators for

their considerations.

This section presents for each recommended project:

• Brief summary of the research project,

• Specific objectives of research,

• Originally submitted budget,

• Recommended revised budget (where applicable),

• Statement of significance to the dairy industry,

• Copy of Director comments to Principal Investigator,

• Overall Director recommendation for WCDPRT funding .

68

•

•

•

Project Title: Production of Extracellular Proteases of Brevibacterium linens for Use in Lowfat Cheese

Principal Investigator: Bart C. Weimer, USU

9. Summary of Proposed Project: Consumers are demanding more lowfat and nonfat foods. Many food ingredient

companies have responded by developing fat substitutes made from protein, starch,

and pectin. The dairy industry has little experience using fat replacers in fermented

milk products.

Addition of fat replacers creates a new dimension to the microbiology of

fermented dairy products. Interactions between fat replacers and dairy cultures are

unknown. Functionality of fat replacers will be effected by the proteinases, peptidases,

and other metabolites the bacteria used during cheesemaking.

Lack of flavor development and hard body in reduced fat dairy products is a

common problem. Highly proteolytic organisms, like Brevibacterium linens, have been

used to accelerate the ripening of full fat cheese. They sometimes cause too much

proteolysis which result in strong flavored and soft bodied full fat cheeses. Use of this

organism in lowfat cheese may produce a product which has an acceptable level of

flavor and body. We will investigate the use of a whey-based medium to produce

enzymes that increase flavor development and body in Cheddar cheese made with fat

replacers .

69

------

4.1. Specific Objectives (State in outline form, objectives of the research and the hypothesis being tested)

Recently hydrocolloid fat replacers have been developed by many food

companies. Some fat replacers may have the capability to replace milk fat in

fermented dairy products. Documentation in the literature is scarce about

• interactions between bacteria used in fermented dairy products and fat replacers.

•

•

Flavor and body development is a common problem in lowfat cheese.

Proteolysis of milk proteins can increase flavor and body in cheese. B. linens is

used to accelerate ripening in full fat products. Use of B. linens may increase the

flavor and soften the body of lowfat cheese.

We propose to develop a whey-based medium for the growth of B. linens. Growth products from B. linens will be harvested and used, with fat replacers, in

determining manufacturering parameters for lowfat Cheddar cheese with

acceptable flavor and body. This will be achieved by completing the following

objectives:

1. Develop a whey-based medium for the production of extracellular proteases by Brevibacterium linens.

. 2. Determine the interaction of the rate of proteolysis from proteases produced by Brevibacterium linens in the presence of fat replacers and coagulating enzymes in milk.

3. Develop processing parameters required to produce Cheddar cheese with acceptable flavor and body using milk fat replacers and proteases from Brevibacterium linens.

1. Proposal Title: Production of extracellular proteases by Brevibacterium linens for use in lowfa t cheese.

July 1, 1992 through June 31, 1994

Salaries Year 1 Year2 Total Graduate Student 6,630 6,962 13,592 Graduate Student 10,000 10,500 20,500

Research Scientist (25%) 10,000 10,500 20,500 Lab Assistant 3,000 3,150 6,150

Fringe Benefits 3,700 3,885 7,585

Subtotal $33,330 $34,997 $68,327

Expenses Supplies 8,500 10,000 18,500

Travel 500 500 1,000 Publication Costs 1,000 2,000 3,000

Amino acid analysis 3,000 3,000 6,000

TOTAL $46,330 $50,497 $96,827

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•

•

4.4. Significance of Proposed Projects to the Dairy Industry (Clearly state how the project is to increase use of milk or dairy products by humans)

Sales of foods perceived as healthy has doubled between 1986 and 1990 and are

expected to double again by the year 1995 (Cheese reporter, Oct 18, 1991). Retail sales of

these foods amounted to $14.3 billion in 1990, up 16% from 1989. Lowfat and low

cholesterol foods accounted for 57.3% of those sales. Dairy products accounted for

34.3% of food retail sales, mostly from frozen yogurt and dairy desserts. Fat substitutes

are predicted to be the largest growth segment of the food ingredient industry in the

1990's (Cheese reporter, Oct 18, 1991). In a recent diet by Updike (The Morman Diet, 1991, CFI, Orem, UT) milk is

described as a health hazard. He wondered why people eat milk because of the fat

content and other safety problems. He reorganized the food groups to exclude dairy

and meat products. Propaganda of this sort is easier to disregard if acceptable lowfat i

products are available in the market place. Research of the type described in this

proposal would be useful to the dairy industry as a means to take advantage of the

rapidly expanding, lowfat food market. While the lowfat food market has been expanding in recent years natural cheese

products are missing in the market. Marketing lowfat dairy products will increase the

milk sales and consumer acceptability of non-traditional dairy products. Development

of more lowfat cheese products requires an understanding the functionality of fat

replacers and their interactions with starter bacteria during cheesemaking and

maturation .

71

•

•

•

The TA C evaluation of this proposal ranked 5th of 14 proposals considered. The T A C comments should be read carefully because there are contradictory interpretations of some ideas presented in the proposal.

The Director suggests the following budget changes:

Personnel Year 1 Year2 Total

Research Scientist (25%) 7,000 7,350 14,350 Graduate Student 10,000 10,500 20,500 Fringe (37 .5 and 38.0% 2,625 2,793 5,418 Laboratory Technician 3,000 3,150 6,150

Expenses (Approved as submitted)

TOTAL $35,625 $39,293 $74,918

The Director recommends funding of this project at the revised budget levels .

72

•

•

•

Project Title: Bacteriophage-Resistance Gene Replacement in Lactococcus lactis

Principal Investigator: Bruce L. Geller, OSU

9. Summary of Proposed Project

Recent work in my laboratory, funded by a grant from the National Dairy Research and Promotion Board, has discovered both a gene and a protein required for virus infections of the bacteria used as starter cultures in cheddar cheese manufacturing. Although we have already isolated virus-resistant strains with mutations in the required gene, the nature of the mutations makes them prone to revert back to the virus-sensitive state. The proposal before you now requests funding to create a virus-resistant strain that would be permanent and stable against reversion back to the virus-sensitive state.

To accomplish this, the project proposes to make a number of permanent, irreversible mutations in the gene already isolated from the on-going project. The natural gene in the bacterial chromosome would then be replaced with a mutated one. Each of these mutants would be evaluated for resistance to virus, cell viability, and starter culture characteristics necessary to make cheese .

1. Specific Objective

The primary objective of the proposed research is: To replace the wild-type chromosomal pip gene with a mutated copy, in order to create a stable, phageresistant strain.

Strategically, the intermediate aims necessary to attain the primary objective are:

a. To make' in vitro mutations in the cloned gene required for phage infections .

b. To insert the mutated copies of pip into the chromosome and exchange it with the wild-type gene.

c. To test the different mutants for viability, phage resistance, and growth characteristics favorable to cheese-making.

The hypothesis being tested is that the cloned gene product can be mutated in such a way that cell viability and growth characteristics favorable to cheesemaking are not compromised, and the resulting mutant is stably phage-resistant .

73

•

•

•

Budgec InformaCion

1. Proposal Title. Bacteriophage-resistance gene replacement in Lactococcus

lac tis

Salaries YL1 :YLZ. n:..2 total

Graduate student stipend 0 0 0 0

Postdoc fellow 0 0 0 0

Research technician 18,000 18,900 19,845 56,745

Lab helper 500 550 600 1,650

Fringe benefits 9,760 10,061 10,367 30,188

Graduate tuition 0 0 0 0

SUBTOTAL $ 28,260 29,511 30,812 88,583

Supplies 9,600 10,560 11,616 31' 776

Travel 0 1,260* 1,323* 2,583

Publication costs 0 500 800 1,300

Other expenses** 2.000 0 0 2,000

TOTAL $ 39,860 41,831 44,551 126,242

*The principal investigator's cost of attending the American Dairy Science Association annual meeting in Logan, Utah in 1991 was $ 1, 250. It is not anticipated that the costs will become less expensive by 1993 .

**The following equipment will be needed for the proposed project:

1. Automatic hand-held pipe tors, Pipetman models P-20, 200, 1000 $ 500. 600. 900.

2. Horizontal slab gel apparatus, submarine type 3. Electrophoresis power supply for slab gel

\

TOTAL $ 2,000.

The expected life-span of the above equipment is about 7-10 years. We anticipate that the equipment would continue to be used on dairy-related projects, even beyond the proposed funding period. Much of the equipment already in my lab was purchased with money from sources other than the Dairy Board, but has been shared gladly with dairy-related projects .

74

•

•

•

4. Significance of Proposed Project to the Dairy Industry.

Phage infection of cheese and other dairy fermentations is the most significant cause of ruined fermented milk products. Because nearly 30 % of the raw milk produced in the United States is used to make cheese, even a small percentage of the milk ruined by phage contamination represents a large quantity of lost milk, as well as lost profits and higher consumer costs. Often the product of failed cheese fermentations must be disposed of entirely, or sold at a small fraction of what high quality products command. Reduction or elimination of this problem would decrease the waste of milk, and through economic means, would increase the consumption by humans.

The proposed research would create new strains of lactic bacteria for starter cultures that are more phage-resistant than currently available. This strain improvement project differs significantly from those in the past, in that it proposes to use molecular genetic engineering techniques to introduce mutational changes not practically possible by conventional strain improvement technology.

The proposed research plans to take what we have learned from our basic research on phage infections and apply it to the development of new strains. The new strains will be tested for their growth characteristics necessary to make cheese. This testing includes the effects of the new strains on milk proteins, such as the casein proteolytic activities and coagulation times. In this way, the the aims of the proposed research are very much in keeping with the focus of the Western Center for Dairy Protein Research and Technology.·

The TAC evaluation of this proposal ranked 3rd of 14 proposals considered. The TAC comments are positive as to WCDPRT funding .

Budget line item changes are made as follows:

Travel costs are set at $1 000/yr maximum. Each year of the three (3) year project will have a travel allotment of $1000. The changes to each year's total budget request are:

Year 1: $40,860 Year 2: $41,571 Year 3: $44,228

TOTAL REQUEST: $126,659

The Director will recommend funding of this project at the new funding levels listed directly above .

75

•

•

•

Project Title: Purification of Monospecific, Polyclonal Antibodies from Bovine Cheese Whey

Principal Investigator: Rodney J. Brown, USU

9. Summary:

Milk is a unique biological fluid that provides complete nutrition to newborn mammals and is

a dietary source of many important nutrients in humans, both young and old. Most of the

constituents in milk have been used for food manufacture, such as cheese and yogurt. Recently,

a market has developed for lactoferrin, an iron binding protein in milk that has implications in

preventing disease. But, many valuable milk constituents have not been exploited for human

use. We propose to develop new technologies for isolation of polyclonal, monospecific

antibodies from cheese whey that could be marketed through medical and pharmaceutical

industries.

4.1 Specific Objectives

Immunochemical techniques have become very important diagnostic tools for the medical

industry. Physicians are using specific antibodies for diagnosing and following the progress of

many diseases. Medical researchers also rely heavily on immunochemical techniques for

researching such diseases as cancer and autoimmune deficiencies. A third area is disease

treatment using antibodies. Our objective is to produce these antibodies in milk and purify them

from cheese whey. This will provide a tool for producing antibodies for any medically important

human or nonbovine antigen.

Relationships between human and bovine species are distant enough to allow antibody

production in cattle against human antigens. Purified, monospecific, human antibodies now

being isolated from rabbit, goat, and sheep's blood could be isolated from cheese whey, in

essence making cattle "biological factories". This approach to producing "biopharmaceuticals"

would overcome problems inherent with drawing blood from animals and provide a large base of

antibody supply for the medical industry. This also would give dairy farmers a new market for

these milk constituents and increase the utilization of milk.

Our specific objectives of this project are to:

1. Optimize antibody titers in bovine milk by testing various immunization schemes.

2. Develop a purification method using affinity chromatography with immobilized antigen' for

purifying antibodies from cheese whey.

3. Develop pilot plant scale-up equipment as a model for full-scale, commercial production of . . 76

antibodies.

•

•

•

SECTION 3 Budget Information

1. Proposed Title: Purification of Monospecific, Poly clonal Antibodies from Cheese Whey

Duration of project: July 1, 1992 through June 30, 1995

Salaries Year 1 Year2 Year3 Total Research Scientist (75%) 30,000 31,500 33,000 94,500

Frin~e Benefits 11,100 11,655 12,210 34,965 Graduate Student 10,000 10,500 11,000 31,500

0 0

Subtotal $51,100 $53,655 $56,210 $160,965

Expenses Year 1 Year2 Year3 Total SuEElies 5,000 10,000 15,000 30,000 Travel I 2,000 4,000 6,000

Publication Costs2 1,000 2,000 3,000 ELISA Plate Reader 10,000 10,000

TOTAL $66,100 $66,655 $77,210 $209,965

I Travel costs are over $1000.00 because many trips to Chappel Cheese in Wayne County, Utah will need to be taken during the pilot plant scale-up. We also are planning to report this research at the ADSA annual meeting after each year. Each trip will cost approximately $1000.00.

2Journal of Dairy Science- $70.00/page

4.4 Significance of Proposed Project to the Dairy Industry

We expect the results from this research to verify our hypothesis that antibodies to specific

antigens can be isolated from cheese whey and purified for use in the medical and

pharmaceutical industries.

Immunochemical techniques have become very important diagnostic tools for the medical

industry. Doctors are using specific antibodies for diagnosing and following the progress of

many diseases. Medical researchers also rely heavily on immunochemical techniques for

looking at such diseases as cancer and autoimmune deficiencies. A third area is in disease

treatment. Our objective is to produce these antibodies in milk and purify them from cheese

whey. Relationships between human and bovine species are distant enough to allow antibody

production in cattle against human antigens. Purified, monospecific, human antibodies now

being isolated from rabbit, goat, and sheep's blood could be isolated from cheese whey, in

essence making cattle "biological factories". This approach to producing "biopharmaceuticals"

would overcome problems inherent with drawing blood from animals and provide a large base of

antibody supply for the medical industry. This also would give dairy farmers a new market for

these milk constituents and increase the utilization of milk .

77

•

•

•

The TAC evaluation of this proposal ranked 7th of 14 proposals considered. The TAC comments are brief but the high evaluation score indicates some broad acceptance.

The Director suggests that project objectives and budget revisions be made. The project should investigate objectives 1 and 2 during a two (2) year project. Objective 3 (design of a pilot plant) should be left for a future proposal pending significant positive results from this project's objectives 1 and 2.

The Director suggests budget revisions be made as follows:

Personnel

Research Scientist (75%) Fringe (37.5 and 38.0%)

Expenses

Supplies Travel Publication Costs ELISA

TOTAL

Year 1

21,000 7,875

5,000 1,000

10.000

$44,875

78

Year2

22,000 8,360

10,000 1,000 1,000

$42,360

Total

43,000 16,235

15,000 2,000 1,000

10.000

$87,235

•

•

•

Project Title: Rheology and Microstructure of Mozzarella Cheese

Principal Investigator: Donald J. McMahon, USU

9. Summary: Consumption of Mozzarella cheese has grown steadily at approximately 10%

per year for the past ten years. The physical properties of Mozzarella cheese, its melt

and stretch, has fueled this growth There have been a number of recent studies

(including those already conducted at Utah State University) of factors that affect

stretch and melt characteristics of Mozzarella cheese but our knowledge of why

Mozzarella cheese stretches and melts is still very limited. Part of this information

will be provided in a study being funded by the National Dairy Board on the

development of culture systems and the use of fat substitutes for the manufacture of

low fat Mozzarella. This proposal requests funding for two years to develop

experimental techniques and equipment to study the microstructure of Mozzarella

chees_e under melting conditions using electron microscopy. Upon successful

completion of this phase of the project, additional funding is sought for an

additional two years to apply these techniques to the study of the physical properties

of Mozzarella cheese.

An understanding of the process of cheese melting would enable us not only

to control the production of conventional Mozzarella cheese but also develop new

cheese products designed to satisfy consumer demands for functional cheeses. The

funding of this project would take the study of Mozzarella cheese to a more basic

level and provide measurements of the stretch and melt characteristics so that

observed changes in physical properties could be explained .

79

·-4.1 Specific Objectives

It is well known that the stretch and melt characteristics of Mozzarella cheese

are important aspects of its appeal to consumers. Although there have been a

number of recent investigations of the factors that affect these properties, our

knowledge of why Mozzarella cheese stretches and melts is still very limited.

An understanding of the process of cheese melting would enable us not only

to control the production of conventional Mozzarella cheese but also develop new cheese products designed to satisfy consumer demands for lowfat cheeses. The

specific objectives of this project are to:

A. Develop combined rheological and electron microscopic techniques for the measurement the melting and stretching phenomena of cheese.

B. Study the protein-fat interactions using the technique developed above to determine how stretch and melt properties are related.


• 1. Proposed Title: Rheology and microstructure of Mozzarella cheese


Item Year1 Year2 Year3 Year4 Total Research Associate $15000 $10000 $10000 $35000 Graduate Student $10000 $10000 $10000 $30000 Frin~e Benefits $0 $5700 $3800 $3800 $13300 Salaries Subtotal $0 $30700 $23800 $23800 $78300 Laboratory Supplies $10000 $10000 $4000 $4000 $28000 EM Time $3000 $2000 $5000 $10000 $20000 Technical Services Time $6000 $3000 $9000 Publications $800 $800 $1600 $3200 Travel $1000 $1000 $1000 $3000 EguiEment Lease/Purchase $15000 $15000 $30000 Su_EElies Subtotal $19000 $16800 $25800 $31600 $93200 Clerical Costs $380 $336 $516 $632 $1864 TeleEhone $190 $168 $258 $316 $932 Accountin~ Costs $380 $336 $516 $632 '$1864 Total $19950 $48340 $50890 $56980 $176160

• 80

• 4.4 Significance of Proposed Project to the Dairy Industry

Per capita consumption of Mozzarella cheese in the U.S. has increased from 0.4 pounds in 1960 to 6.4 pounds in 1989. Consumption of Italian-type cheese has soared 48 percent since 1985, with Mozzarella (57%) and Ricotta (36.4%) cheeses leading in this market segment. The continuing success of pizza has been the major factor in the growth and popularity of Italian cheeses. Even with this type of growth there are still a number of areas in cheese sales that could be filled with a lowfat or nonfat Mozzarella cheese. An understanding of the effects of modifying fat content or other parameters in Mozzarella cheese on the microstructure of the cheese is needed to evaluate methods for retaining the critical physical properties of this type of cheese. Until this is achieved the production of Mozzarella will remain on a trial and error basis.

The TAC evaluation of this proposal ranked 12th of 14 proposals considered. The TAC comments and questions whether the research could be done in a more streamlined fashion. The WCDPR T can fund the necessary amount to combine the main thrust of this

• proposal with the lowfat and nonfat research awarded by the NDPRB Competitive Research Program.

•

The Direct~r .suggests that yo':~ resubmit the necessary infonnation showing a linkage to the Competmve Research proJect. You should also submit a revised budget that indicates the minimal WCDPRT input into the linkage research .

81

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•

Project Title: Function of Whey Proteins and Lactose in Age Gelation of UHT-Processed Milk Concentrate: Part 2


SUMMARY

The mechanism by which age gelation in UHT sterilized milk concentrates occurs is still unknown.

There have been many factors implicated and at best an empirical approach is taken to extend shelf

life of sterilized milk products. UHT sterilization promotes association between K-casein and

~-lactoglobulin. In milk concentrates the concentration of whey proteins and lactose are increased.

Their role in the age gelation process will be studied in this project

.JUSTIFICATION

The overwhelming success of dairy production in the United States has created a situation that calls

for more attention to milk utilization and marketing. Development of new dairy products has taken

precedence over production of more milk .

International markets for U.S. dairy products could be developed if attention could be directed to

manufacture of stable products from our surplus dairy production. The competitive position of the

U.S. would be enhanced by new and better quality products and our surpluses of dairy

commodities would be reduced. Production of dairy products other than powdered milk, butter

and cheese with long shelf lives should be a major priority. Specifically, for this project, a way of

producing rehydratable milk concentrate which will not gel before reaching the intended consumers

is needed to make U.S. Dairy products more widely available on the world market. The major

limitation to production of such a product is irreversible gelation when exposed to higher-than

refrigerator temperatures over a long period of time. The gelling phenomena must be understood

and means devised to prevent gel formation before rehydratable milk concentrates made from

surplus U.S. milk can be sold abroad .

82

•

•

-----y.y,~~---------------------------------------------------SPECIFIC OBJECTIVES

The specific objectives of this project are: ·

1 . To determine the influence of lactose concentration of milk concentrates on age

gelation ..

2. To determine the role of whey proteins (especially f)-lactoglobulin) in the mechanism of

age gelation of UHT sterilized milk concentrates.

· 3. To monitor changes in casein micelle structure that occur during storage of UHT

sterilized milk concentrates and relate this their stability.

From the partial funding already provided by the Western Dairy Foods Research Center,

Objective 1. has already been completed. It was determined that lactose has plays neit!Ier a

protective nor detrimental role in age gelation. From our observations we concluded that

proteolysis has a significant effect on causing age gelation and that protein-protein

. interactions at high storage temperatures can interfer with the physico-chemical changes of

casein micelles that results in gelation. Further data will have been collected on Objective 3

by June 1992. This funding request is to complete Objective 2 using differential scanning

calorimetry and immuno-gold labeling electron microscopy .

BUDGET INFORMATION

Item Year1 Year2 Total Visiting Professor $8000 $8000 Graduate Student $10000 $5000 $15000 Salaries Subtotal $10000 $13000 $23000 Laboratory SU££lies $6000 $7000 $13000 EM Time $4000 $4000 $8000 Publications $500 $500 $1000 Travel $1000 $1000 $2000 Supplies Subtotal $11500 $12500 $24000 Clerical Costs $230 $250 $480 Accounting Costs $230 $125 $355 Telephone Costs $115 $125 $240 Total $22075 $26000 $48075

Project 15. Function of Whey Proteins and Lactose in Age Gelation ...

The TAC evaluation of this proposal ranked 4th of 14 proposals considered. The TAC, comments are sketchy but the overall high evaluation score indicates broad acceptance of the project concept Your previous clarification of the "Visiting Professor" budgeted line item has met with NDPRB approval.

• The Director will recommend funding of the project at the requested levels of $22,075 for 1992-93 and of $26,000 for 1993-94.

83

Project Title: Extrusion Processing of Whey Proteins (Pending)

• Principal Investigator: Conly L. Hansen, USU

•

•

SUMMARY OF PROPOSED PROJECT

Annual production of whey in the United States is more than 50 billion pounds. Over 50 percent of this production is still disposed as raw waste (Anon, 1991). This whey is highly polluting if discharged untreated and represents a serious environmental problem. Whey has a chemical oxygen demand of approximately 78,000 ppm, an indication of very high pollution potential (Williams, 1984). Disposal of whey as waste is not only costly but also represents loss of valuable proteins and other nutrients. Proteins can be recovered from whey by ultrafiltration or by pH adjustment. Annual availability of whey proteins in the U.S. is 500 million pounds. Extrusion technology has the potential of restructuring whey proteins and imparting unique characteristics to them for use in fabricated foods. Development of extruded whey protein food products will create new markets for whey proteins. The restructured whey proteins can be used as a nutritive ingredient in low fat products where texture is an important quality attribute.

Extrusion processing is done in a food extruder made up of a flighted single- or twin- screw that rotates in a tightly fitting stationary barrel. The process involves forcing moistened proteinaceous and/or starchy food materials to flow under a variety of process conditions and through a specially designed die opening at a predetermined rate. During this process the food ingredients are subjected to a number of unit operations (i.e. mixing, shearing, cooking, forming, texturizing, puffing, and drying) in a single energy efficient, rapid and continuous process (Harper, 1981). Extrusion cooking combines heat and mechanical shear to form a continuous viscous dough by destroying the native organized structure of the macromolecules of food ingredients. As explained by Harper (1986) "The laminar flow within the channels on the extrusion screw and die aligns the large molecules in the direction of flow, exposing bonding sites which leads to crosslinking and a reformed, expandable structure that creates the crunchy or chewy texture in fabricated foods." ·

NMProtein HYDRATION

CROSHINKS

~ SETliNG

_:;

,9~ UNFOLDING

!''"" ..__~ COOLING ~

ALIGNING SlliCTCHING

Schematic representation of protein restructuring process (Shen and Morr, 1979; Aguilera and Stanley, 1986)

84

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•

•

-Whey protein concentrate is produced commercially by ultrafiltration. Globular proteins

constitute 68% of whey proteins (Kosaric and Asher, 1985). The overwhelming success in applying extrusion technology to globular soy protein suggests the potential of restructuring whey protein by extrusion. The proposed project will determine operating parameters for extruding whey proteins to alter their characteristics so that they will be more widely used in fabricated food products. The focus of the project is development of new value-added, nutritional food products which will expand the horizon of whey proteins to nontraditional markets. The specific objectives are:

!.Determine effects of feed composition and process parameters including temperature, pH, screw speed, and extruder length/diameter (L/D) ratio on physical, chemical and functional characteristics of whey proteins extrudate.

2.Determine effects of moisture content of whey protein feed and process parameters on microstructure of whey proteins extrudate.

3.Investigate co-extrusion of whey protein with other proteinaceous and starchy ingredients.

4.Determine product applications of the high protein extrudate and co-extrudate in order to find new markets for whey proteins.

A billion pounds of protein are thrown into the ocean annually by Alaska's fishing and processing industries alone (Goldhor, 1990). This pollutes the receiving waters and is a loss of valuable protein. Despite its high nutritional value, by-product proteins remain largely unused for human consumption due to a lack of technology to transform the material into acceptable food products with desirable quality attributes. Per capita seafood consumption in the U.S. will increase by approximately 5 pounds by the year 2000. Each pound of increase requires the harvest of an additional 750 million pounds. Any increase in fish catch will result in oversupply of materials available for by-product use. Muscle proteins recovered from by-products does not have good textural attributes which limits market value. Mixtures of mechanically deboned fish protein and globular soy protein have been successfully extruded with improved textural attributes. Specific textural and nutritional characteristics have been imparted to the products by regulating appropriate composition and process variables (Murray and Stanley, 1980; Bhattacharya et al., 1988, 1990). These studies suggest the potential of applying extrusion technology to produce value-added food products from mixtures of fish and globular whey proteins. Manufacture of new value-added food products will create new opportunities to market whey proteins and will increase the income from milk made into cheese.

The proposed project is in the areas of thermal processing, whey utilization, and production of value added products. These areas have been identified as "Research Areas of Interest" by Wester Center for Dairy Protein Research and Technology .

85

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2.1 Specific objectives

Extrusion processing has found widespread application in the food industry for restructuring proteinaceous and starchy material into a variety of fabricated, cooked, and shaped products of varying texture. A myriad of fabricated, cooked, flavored and shaped products are manufactured using food extruders. Cooking extruders are used to produce breakfast cereals, noodles and pasta, biscuits, snack foods, confectionery, chewing gum, modified starches, dried soups, pet foods, crackers, bread crumbs, dried beverage powders, and texturized proteins (Hauck, 1980). The popularity of extrusion cooking stems from the flexibility it offers to produce food commodities of desired nutritional balance in an appealing form. The process is energy efficient, has low production cost, handles a variety of raw ingredients and processing conditions, offers better ingredient utilization with no or very little effluent, and is more versatile than other thermal processing methods (Harper, 1990). The high-temperature-processing capability of extruders with short residence times has been termed HTST processing. HTST processes have been beneficially used to denature enzymes, inactivate antinutritional factors, kill microorganisms and improve digestibility (Mustakes et al., 1964; 1970).

The major research in protein foods has been to find new sources to feed the world's growing population. Converting protein into foods that will be readily accepted has become important only in recent years. Globular proteins have been successfully texturized using food extruders. Textured soy protein is widely used as an extender of red meat products (Maurice et al. ,_ 1976). With recent developments in twin-screw extrusion it is possible to develop product specific characteristics in food extruders. Globular proteins constitute 68% of whey proteins (Korsaric and Asher, 1985). The overwhelming success in restructuring globular soy protein suggests the potential of modifying whey proteins by extrusion to develop product-specific characteristics. The primary objective of the proposed project is to develop extrusion technology for restructuring whey proteins for use in engineered foods. The specific objectives are:

!.Determine effects of feed composition and process parameters including temperature, pH, screw speed, and extruder length/diameter (L/D) ratio on physical, chemical and functional characteristics of whey proteins extrudate.

2.Determine effects of moisture content of whey protein feed and process parameters on microstructure of whey proteins extrudate.

3.Investigate co-extrusion of whey protein with other proteinaceous and starchy ingredients.

4.Determine product applications of the high protein extrudate and co-extrudate in order to find new markets for whey proteins .

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2.4 Significance of Proposed Project

In the dairy industry for every 100 pounds of whole milk processed into cheese, about 90 pounds of whey is produced which contain 5% lactose, 1 % protein and 1% mineral (Morris, 1979). Despite its high nutritional value, a significant portion of the whey remains unused for human consumption due to a lack in economically viable technology to transform whey into acceptable food products with desirable quality characteristics.

The demand for convenience-type foods that combine nutrition with desirable sensory attributes is increasing rapidly. American consumers are increasingly aware of the nutritional importance of proteins, vitamins and minerals, and are avoiding foods containing excess calories and saturated fats. Production of whey protein concentrate by ultrafiltration is an established segment of the dairy industry. Annual availability of whey proteins in the U.S. is 500 million pounds. Proteins recovered from whey can be used in food products. But foods without acceptable appearance, flavor and texture will not be successful, regardless of nutritional quality. The challenge, therefore, lies in fabricating foods that combine nutrition, desirable sensory attributes and convenience. Extrusion processing offers a unique method of producing nutritious foods that will be readily accepted. Whey proteins can be restructured by extrusion for use in fabricated food products. This versatile method has found widespread application in the food industry for restructuring proteinaceous and starchy materials into a variety of fabricated, cooked and shaped products of varying texture. The extrusion process when developed will expand the utilization of milk products in food and will generate additional income from milk used to make cheese.

A.billion pounds of protein are thrown into the ocean annually by Alaska's fish processing industries alone (Goldhor, 1990). This includes by-product proteins from fillet, surimi and canning operations, and fish that are too small or soft for fillet operation. Minced fish have been recovered from these sources. The product lacks textural characteristics and therefore has low value. The United States has the world's largest flatfish resource - 1.1 million pounds a year in the Gulf of Alaska alone (Anon, 1989). Arrowtooth flounder (Atheresthes stomias) which is about half of this resource has limited market because of softer texture compared to other fish specie. This species remains underutilized due to a lack in technology to transform the muscle proteins into food products with acceptable textural attributes. Extruded products with improved texture have been produced from mixtures of mechanically deboned fish protein and globular soy protein. Specific textural and nutritional characteristics have been imparted to the products by regulating appropriate composition and process variables (Murray and Stanley, 1980; Bhattacharya et al., 1988, 1990). Since globular proteins constitute 68% of whey proteins, value-added food products can be produced from mixtures of fish and whey proteins. Manufacture of such fabricated foods will lead to development of new markets for whey proteins.

Raw materials cost in the food industry may account for up to 75% of the cost of processed foods compared to 40% for the chemical industry. This makes efficient raw material utilization an economic necessity. Extrusion processing of whey protein for enhanced use in fabricated

food products w~ll improve the economic viability of the dairy industry by increased utilization of mil~. . The mcome fro~ milk will be significantly increased by bringing the presently underutlhzed or unu.sed portions of milk made into cheese into the human food chain. The proposed research wtlllay the groundwork for further studies on extrusion of UF milk retentate obtained from surplus dairy production and help develop an international market for U.S. dairy products.

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• • • SECTION 3 Budget Information

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Amount Requested From Western Center for Dairy protein Research and Technology

l Salaries I Year 1 I Year 2 I Year 3 I Total I Postdoctoral Fellow: 18,000.00 18,900.00 19,845.00 56,745,00 (U of A - Full Time)

Fringe Benefit (U of A) 5,256.00 5,519.00 5,795.00 16,570.00

Technician or graduate student 12,000 12,600 13,230.00 37,830.00 (USU - part time)

Laboratory Assistant 2,000.00 2,000.00 2,000.00 6,000.00

Q Fringe Benefit (USU) 4,080.00 4,284.00 4,498.00 12,862.00

Subtotal 41,336.00 43,303.00 45,368.00 130,007.00

Supplies 4,000.00 4,000.00 4,000.00 12,000.00

Travel (Including travel & lodging (3-4 months 10,000.00 7,500.00 7,000.00 24,500.00 in Alaska) for the P.I.) See letter attached

Publication Cost 1,000.00 1,500.00 2,500.00

Electron Microscopy 2,500.00 2,500.00 5,000.00

Total 57,836.00 58,303.00 57,868.00 174,007.00

Amount To Be Contributed by U of AK, Fishery Industrial Technology Center

Equipment Rental (Research Extruder) 20,000 20,000 20,000 60,000

The following is Board opinion of the proposals submitted:

• Extrusion Processing of Whey Proteins /Hansen

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The NDPRB Industry Advisory Committee's reaction to this project was not positive as they felt industry would pursue it if there was a need. And further, there is nothing novel here. That, coupled with the fact that most of the dollars for this project will be spent at a university with no dairy interests makes it difficult to approve. As per conversation with Dr. Hansen, though, I will make a few telephone calls before making a final decision.

Jrnsen I Extrusion Processing of Whey Prot~ ins . I T{le following has been discussed with Dr.~: Hansen: 1. lack of a twin-screw cooker ~~truder in the Utah and Western state area, 2. the strong need to minimize ftr,ding, particularly travel, outside the c ntinental United States to a university ~th no dairy interests, and 3. desirability of using skim milk powder in preference t~ whey. The Board agrees to reconsider a~ proposal that addresses issues 2 and 3.

I I

The TAC evaluation of this proposal ranked 11th of 14 proposals considered. The TAC comments indicate that questions arise as to the economic viability of the project and whether the project wi~ "lead to the sale of more milk" These comments are sufficiently strong (as you can seem the attached comments sheets) that greater justification will be required.

I understand that you are also addressing two (2) issues raised by NDPRB (per the June 16, 1992 communication from Barbara Blakistone). All of these issues (NDPRB and TAC) should be discussed in sufficient detail in order to justify the project.

One sol~tion is to scale down the objectives and time of the project. For example, a oneyear pr?Ject could show the applicability of using milk proteins (skim milk and whey) in producmg an acceptable extruded product, including a brief economic analysis of the impact the milk proteins play in the product. If these items (product and cost analysis) . prove successful, another longer-term project can be implemented-next year to really look at the technology.

The Director will not recommend full funding of the project as it now stands. Clarification of the key issues as presented by NDPRB and by TAC can be made and will be considered .

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Project Title: Effects of Iron Fortification on Chemical, Physical, Microbiological and Nutritional properties of Yogurt


--, --~------------ -------- ----------

9. Project Summary:

Consumption of yogurt is primarily by women, children and teenagers as a

luncheon food. However, it is these very populations while having high calcium

requirements are also frequently deficient in iron. Yogurt is an excellent food high in

calcium but lacking in iron. Iron-fortified yogurt can meet both the iron and calcium

needs of these populations and could be promoted as such.

Yogurt has the characteristics of high acidity and distinct flavor which make it

suitable for iron fortification. Its acidity will maintain iron in a soluble state for

absorption and its strong flavor will mask many significant low-grade off flavors that

may t;>e produced by iron. Furthermore, much of the yogurt consumed contains fruit,

the ascorbic acid of which can enhance absorption of the fortified iron.

Understanding how iron is bound in yogurt is essential to developing high

quality products with superior iron bioavailability. We will determine if the iron is

bound to the casein micelle and, if so, where, i.e., internally or on the surface. We will

determine if significant quantities of the fortified iron is bound to the fat globule

membrane or elsewhere. By elemental filtering electron microscopy, we can

determine where the fortified iron is bound. We will determine the responses of the

culture organisms to iron fortification, and we will optimize the iron levels and forms to

produce high quality yogurt.

· 4. · Si~nificance of Proposed Project to the Dairv Tndustrv

It is anticipated that fortifying yogurt with iron will be of value to the dairy industry in two ways; it could increase the amount of milk sold and provide another valueadded product for conversion of fluid milk.

I

Iron-fortified yogurt will be a new dairy product that can provide an effective means of combating iron deficiency particularly in women, young children, teenagers, and elderly people. The low pH of yogurt can enhance the absorbability of the iron

• and decrease any off flavors caused by auto-oxidation. ·

Costs for fortifying yogurt with iron are small. This means that iron-fortified yogurt could be promoted and sold at a significantly increased margin thereby further increasing the value of fluid milk. 90

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Objective A To produce low-fat and non-fat iron-fortified yogurt using FeCls. Fe-casein or Fe-w hey complex as the iron sources

Yogurt fortified with different iron sources (FeCI3, Fe-casein or Fe-Whey protein complex) will be made. Pasteurized and homogenized milk (skim and 2%) will be heat treated at 82°C for 30 min and 6% sugar, .04% stabilizer and NFDM will be added to the hot milk. The milk blend will then be cooled to 43-44°C. The iron sources and yogurt cultures (L. delbruekii ssp. bulgaricus and S. salivarius ssp. thermophilus) will be added to the milk blend and placed in a 45°C incubator until appropriate pH is reached. The yogurt will then be placed in a cold room and allowed to equilibrate for 4-5 days. Samples will be drawn at various times in the culturing, equilibration and storage processes as required in order to follow microbiological, chemical and quality changes throughout the manufacturing process.

Objective B To determine growth and viability of Lactobacillus delbruekii ssp. bulgaricus and Streptococcus salviarius ssp.thermophilus in iron fortified yogurt

It is important to determine if iron fortification affects growth and viability of lactic acid bacteria during yogurt culturing. Iron may enhance growth of some organisms while inhibiting that of others. The total numbers of viable organisms will be determined by counting an Elliker agar. Iron-fortified yogurt will be diluted in autoclaved 0.1% peptone (Richardson, 1985). One-tenth milliliter of each dilution will be spread over agar plates. The plates will then be incubated in an anaerobic environment (BBL Gas Pak, Becton Dickinson Microbiology System Cockeysville, MD) for 48 h at 37°C (Speck and Geoffrion, 1980; Hamann and Marth, 1984). Starter culture population will be monitored after 1, 10, 20, or 30 days of storage at 4°C. MRS agar (DeMan et al., 1960; Speck, 1976; Bracquard, 1981 ), Rogosa agar (Hamann and Marth, 1984) and Elliker agar (EIIiker et al., 1958) will be used to enumerate total numbers of starter bacteria. Rogosa agar and MRS agar will give the number of L. delbruekii ssp. bulgaricus. Therefore, the total number for S. salivarius ssp. thermophilus will be determined by subtracting the number of L. delbruekii bulgaricus from the total number of bacteria (Hamann and Marth, 1984).

Objective C To determine iron binding sites using Elemental Filtering Electron Microscopy

Where fortified iron is bound in dairy products is unknown especially in high acid foods. This information is important for predicting shelf stability of yogurt from microbiological and chemical oxidation perspectives. Iron that is tightly bound to proteins may not be available to culture or spoilage organisms and would not affect their growth. However, the bioavailability of this iron to humans would be high because of their proteolytic digestive enzymes unless it is bound to proteins that"'have poor gastrointestinal digestibility.

Elemental Filtering Electron Microscopy (EFEM) will be used to determine iron binding sites (e.g., casein micelle, surface of fat droplets, etc.). EFEM eliminates all

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inelastically scattered electrons, therefore, a spectroscopic image of the selected element (e.g., iron) can be constructed through electron energy-loss spectrometry. The element-spectroscopic image will enable us to locate the binding site or sites where the iron is bound .

Samples with various iron sources will be prepared as follows: yogurt and 50% glutaraldehyde will be mixed in a test tube. The mixture will first be incubated at room temperature for 1 0 min and then will be transfered to a 45°C water bath. The mixture will then be poured into a petri dish and allowed to solidify (alternatively, if the yogurt does not solidify properly with glutaraldehyde alone, it can be mixed with 3% agar and allowed to solidify at 45°C. Using a razor blade, the gel will be cut into small strips (.5 em length, 1-2 mm width). The small strips will then be placed in a vial containing 2.5% glutaraldehyde in phosphate buffer.

The sample will then undergo an additional fixation with osmium tetraoxide (Os04). Os04 acts as both fixative and electron stain. It covalently binds to unsaturated fat and phospholipids thereby stabilizing the fat structure.

After fixation with glutaraldehyde and Os04, the sample will undergo a dehydration step which involves gradual removal of water by dehydrating with alcohol or acetone and replacing all free water with a fluid that is miscible with both water and the embedding medium. The dehydration will be done gradually to prevent osmotic shock to the protein structure.

The sample will then be gradually treated with propylene oxide which acts as a transitional fluid because it is completely miscible with both alcohol and the embedding medium (Youssef, 1984). The propylene oxide is then replaced with embedding medium through infiltration. The embedding medium is a Standard Mollinhauer Araldite Mixture (EPON) which polymerizes upon heating at 60°C.

Objective D To determine the best procedure for making a high -quality ironfortified yogurt

The quality of iron-fortified yogurt will be determined by analyzing its chemical and physical properties, and conducting sensory evaluations.

Chemical Analysis:

I. Iron Analysis The ferrozine method (Carter, 1971; Stookey, 1970) will be used to quantitate

iron in the yogurt after complete wet-ashing with nitric acid and H202. Ferrozine reacts with the ferrous iron and produces color. A Beckman DU-8 spectrophotometer will be used to measure absorbance at 562 nm (Zhang and Mahoney, 1989).

II. Thiobarbituric Acid Test Lipid oxidation materials in yogurt will be quantitated as malondialdehyde using

a distillation method (Tarladgis et al., 1960). Beckman Du-8 spectro-photometer will be used to measure absorbance, 538 nm, of the thiobarbituric acid reactive substances .

92

Sensory Analysis:

• Yogurt quality will be evaluated for appearance, texture, oxidized off flavor, and

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yogurt flavor by about 10 dairy experts trained to detect iron-induced oxidized off flavor and metallic flavor.

Yogurts developed which give favorable sensory results in the expert panel will be further evaluated using lay panels with about 100 participants. The hedonic scale method will be used on all samples. Panelists will be asked to rate samples for taste, texture, appearance, preference and willingness to purchase. Data will be analyzed against frequency with which the panelists consume yogurt and their familarity with yogurt flavor.

Objective E To determine the bioavailability of iron in iron-fortified yogurt

To ensure that the iron is absorbable from the fortified yogurt, it is necessary to determine its absorption in animals. Iron-deficient animals will be used to determine its maximal absorbability and normal animals will be used to ensure that excess iron will not be absorbed since iron absorption is regulated by body iron stores. Irondeficient subjects requiring additional iron will absorb lots of iron and return quickly to normal iron status, while normal subjects with high iron stores should absorb very little iron ~rid avoid toxicity. We will use the animal methods described by Zhang and Mahoney (1989) who used this strategy for evaluating bioavailability of iron-fortified cheese. They determined the percentage of the iron consumed which was incorporated into hemoglobin, hemoglobin regeneration efficiency .

1. Proposed Title; Effects of iron fortification on chemical, physical, microbiological and nutritional properties of yogurt

Duration of project: May 1, 1992 through Apr 30, 1995

Salaries Year 1 Year2 Year 3 Total Sharareh Hekmat, MS. 12,000 12,600 13,230 37,830

Expenses Year1 Year2 Total Supplies 3,500 4,500 3,300 11,300

Travel 1,000 1,000 1000 3,000 Publication Costs 200a 900b 1 ,800b 2,900

~-(Electron Microscop~} 10,000 10,000 20,000 Total Budget Reguest $26,700 $29,000 $19,330 $75,030

a: visual preparation for ADSA meetings. b: visual preparation for ADSA meetings plus manuscript publication in JDSA, 1 paper year 2 and 2 papers year 3 .

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lli. Mahoney's project "Effects of Iron Fortification on Chemical, Physical, Microbiological, and Nutritional Properties of Yogurt" lists you as Co-Principal Investigator. As such, this memo informs you that I will request funding of this project by the WCDPRT at next week's OAC meeting.

The TAC evaluation of this proposal ranked 1st of 14 proposals considered. The TAC comments are included. The comments require only some minor clarification; the overall review of the proposals appears highly acceptable and worthy of funding.

I request the that this project be limited to two (2) years to investigate objectives A, B, C, and D only. Thesse objectives investigate product quality and process parameters in the manufacture of the yogurt product. Nutritional studies should be left to investigate if these four (4) objectives prove successful.

I recommend the proposed budgets for year 1 ($26,700) and year 2 ($29,000) be used to research objectives A, B, C, and D of the proposed project. The recommendation will include the effective dates of the revised project to be July 1, 1992 to June 30, 1994 .

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Project Title: Effects of pH, Salts, and Thlolation on Thermal Gelation of Caseins Alone and in Mixed Protein-Protein and Protein

Polysaccharide Gels, R.lu.i

Interactions Between Milk Proteins, Starter Cultures, and Hydrocolloidal Milk Fat Replacers, R.lu.i

Interaction Between Milk Proteins During Thermal Processing and Its Effects on the Physicochemical Properties of Dairy Foods

Principal Investigator: Rodney J. Brown and Bart C. Weimer, USU

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9. Summary of Proposed Project: Bovine caseins contribute excellent nutritional value to food products but their principal

value in most foods is their functional contributions. Use of caseins and caseinates in food formulations as functional proteins is increasing, eg., cheese analogues, meat products, coffee creamers, dietary formulations, etc. Gel forming ability of proteins is an important functional property in food systems. The ability of gels to act as a matrix for holding water, lipids, sugars, flavors, and other ingredients is useful in food applications and in new product development. Because of the high proline and low-sulfur amino acids content, caseins have a random coil structure with a low percent helix orB-sheet and as a consequence, it was thought that caseins do not form heat-set gels. Hence, thermal gelation of caseins received very little attention. However, limited research done in the past shows that B-CN and as-CN can form highly ordered gels at low protein concentrations in acid media (by means surface contacts only) without any conformational changes. Formation of reversible gels with moderate heating (50-60°C) has been observed in calcium caseinate dispersions which were formulated with high concentrations of protein and calcium. On cooling, the gel reliquefied. This type of gel is rare among globular proteins and is only possible because of the unique structural features of caseins, namely, random coil structure and the presence of ionic, hydrophobic, and ester phosphate groups in the polypeptide chain.

Thus, the rare gelation phenomenon of caseins needs further research to fully understand the mechanism of gelation at different pHs and salt concentrations and rheological properties of these gels. Also, a thorough understanding of the performance of caseins in mixed protein-protein, and protein-polysaccharide gels is needed for their effective utilization in food formulations. It is particularly important because of the increased use of polysacchrides in low-fat foods as fatreplacers. Therefore, this study will provide new insights into the heat-set gelation of caseins and most importantly, open up new avenues for the use of caseins in fruit-based gels under acid conditions, increased utilization of caseins (and milk) in new food formulations and for the preparation of new casein-based food ingredients for use in low-fat foods, and hence increased consumption of milk-based products by humans ....... , " ·' .,.. '· ·· ~ · · · 4.1 Specific Objectives

Because of the high proline and low-sulfur amino acids content, caseins have a random coil structure with a low percent helix or B-sheet and as a consequence, it was thought that caseins do not form heat-set gels. Hence, thermal gelation of caseins received very little attention. However, limited research done in the past shows that B-CN and as-CN can form gels in acid media (by means surface contacts only) without any conformational changes. Also, formation of reversible gels (gel reliquefies on cooling) with moderate heating (50-60°C) has been observed in calcium caseinate dispersions. This type of gel formation is rare among globular proteins and is only possible because of the unique structural features of caseins, namely, random coil structure and the presence of ionic, hydrophobic, and ester phosphate groups in the polypeptide chain.

Hypothesis. The rare gelation phenomenon of caseins needs further research to fully understand the mechanism of gelation at different pHs and salt concentrations and rheological properties of these gels. Also, a thorough understanding of the performance of caseins in mixed protein-protein, and protein-polysaccharide gels is needed for their effective utilization in food formulations. It is particularly important because of the increased use of polysacchrides in low-fat foods as fat-replacers. Therefore, this study will provide new insights into the heat-set gelation of caseins and most importantly, open up new avenues for the use of caseins in fruit-based gels under acid conditions, increased utilization of caseins (and milk) in new food formulations and for the preparation of new casein-based food ingredients for use in low-fat foods, and hence increased consumption of milk-based products by humans. Therefore, the specific objectives of this project are to:

' 1. Study the gelation of caseins (<Xsl-, <Xs2-, B- and K-) in water and buffer systems as a

function of different protein concentrations, temperatures, pHs, divalent cations (eg., ca2+), and structure stabilizing (Cl-, S04=) and structure destabilizing (SCN-, Cl04-) anions .

2. Study thiolation (covalent attachment of new -SH groups) of caseins on thermal gelation.

3. Study mixed casein-protein gelation with whey proteins, egg proteins, meat proteins, and gelatin. 96 '

4. Study mixed casein-polysaccharide gelation with K-carrageenan, alginate, and pectin.


1. Proposed Title: Effects of pH, Salts and Thiolation on Thermal Gelation of Caseins Alone and in Mixed Protein-Protein and Protein-Polysaccharide Gels.

• Duration of project: July 1, 1992 through June 30, 1995

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Salaries Year 1 Year2 Year3 Total Dr. Brown, Professor (5%) NoSal~ NoSal~ NoSal~ NoSal~ Dr. McMahon (10%) No Salary No Salary NoSal?JY No Salary Dr. Redd~, Research Asst. Prof. (100%) 36,000 37,800 39,690 113,490 Frin~e Benefits 13,320 14,175 15,083 42,578

Subtotal $49,320 $51,975 $54,773 $156,068 Expenses Year 1 Year2 Year3 Total SuEElies 7,000 4,000 3,000 14,000 Travel 1,000 2,000 2,000 5,000

Publication Costsl 500 1,000 1,500 3,000 -Other Expenses

Bohlin VOR Rheometer (lease) 10,000 10,000 10,000 30,000

TOTAL $67,820 $68,975 $71,273 $208,068

1 Proposed journal(s ) to which the project results/manuscripts will be sent and the corresponding

page charges: J. Dairy Sci., $70 per printed page .

4.4 Si~nificance of Proposed Project to the Dairy Industry

Bovine caseins contribute excellent nutritional value to food products but their principal value in most foods is their functional contributions. Use of caseins and caseinates in food formulations as functional proteins is increasing. Gel forming ability of proteins is an important functional property in food systems. The ability of gels to act as a matrix for holding water, lipids, sugars, flavors, and other ingredients is useful in food applications and in new product development Because of the high proline and low-sulfur amino acids content, caseins have a random coil structure with a low percent helix or B-sheet and as a consequence, it was thought that caseins do not form heat-set gels. Hence, thermal gelation of caseins received very little attention. However, limited research done in the past shows that B-CN and as-CN can form highly ordered gels at low protein concentrations in acid media (by means surface contacts only) without any conformational changes. This type of gel is rare and is only possible because of the unique structural features of caseins, namely, random coil structure and the presence of ionic, hydrophobic, and ester phosphate groups in the polypeptide chain.

Thus, the rare gelation phenomenon of caseins needs further research to fully understand the mechanism of gelation at different pHs and salt concentrations and rheological properti~s of these

·gels. Also, a thorough understanding of the performance of caseins in mixed protein-protein, and protein-polysaccharide gels is needed for their effective utilization in food formulations. It is particularly important because of the increased use of polysacchrides in low-fat foods as fatreplacers. Therefore, this study will provide new insights into the heat-set gelation of caseins and most importantly, open up new avenues for the use of caseins in fruit-based gels under acid conditions, increased utilization of caseins (and milk) in new food formulations and for the preparation of new casein-based food ingredients for use in low-fat foods, and hence increased consumption of milk-based products by humans.

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4.1 Specific Objectives

In response to consumer demand, hydrocolloid fat replacers have been developed by

many food companies. Some of these may have properties required to replace milk fat in dairy

products. We hypothesize that fat replacers interact with milk proteins and dairy starter cultures

helping to give fat-like characteristics to lowfat dairy products. To test this hypothesis, we will

study interactions of milk protein, dairy starter cultures and milk fat replacers that are claimed to

have milk fat mimetic properties. Our specific objectives are to:

1. Examine coagulation, protein binding, and stability of milk proteins in the presence of fat replacers

a. Quantify the amount of milk protein binding to fat replacer particles.

b. Examine the rennet and acid coagulation time and curd firmness in milk with fat removed and fat replacers added.

c. Determine the partitioning of fat replacers between curd and whey during coagulation and syneresis.

d. Check heat and pH stability of fat replacers in the presence of milk proteins.

2. Stability of fat replacers in dairy systems during growth of lactic starter cultures

a. Monitor growth and acid production rates of lactic acid bacteria in milk with fat replacers.

b. Measure the amount of enzymic (proteinase, peptidase and glucosidase) breakdown of milk proteins and fat replacers by lactic acid bacteria.

c. Determine the manufacturing time and the amount of starter culture required to produce lowfat cheese containing fat replacers .

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• 0 bjective 1. Examine coagulation, protein binding, and stability of milk proteins in the presence of fat replacers

a. Quantify the amount of milk protein binding to fat replacer particles.

The quantity of milk proteins adsorbed to the fat replacer particles will be determined by

homogenizing the particles into skim milk of known protein concentration. The particles and

bound protein will be removed by centrifugation and the serum analyzed for protein content.

The protein load (mg protein/ m2 surface area) will be calculated using particle surface area data

obtained with the Malvern Master Sizer laser light scattering instrument. The serum also will be

analyzed by electrophoresis or liquid chromatography to determine if individual milk proteins

were depleted during the homogenization processindicating preferential binding to the particle

surface. This data will be compared to data obtained when the same experiments are performed

using milk fat. Particle size distributions will be determined using a Malvern Master Sizer laser

light scattering instrument. Simplesse®, StellarTM, Paselli SA2®, and Slendid® will be analyzed

and compared to milk fat globule size distribution determined on the same instrument.

b. Examine the rennet and acid coagulation time and curd firmness in milk with fat removed and fat replacers added.

Milk will be obtained from the University Dairy Farm and skimmed by centrifugation. Milk

prote~n will be reconstituted with milk fat and fat replacers so that whole milk, 25% reduced fat

milk, 50% reduced fat milk, 75% reduced fat milk and skim milk are prepared. Rennet and acid

coagulation characteristics of these samples will be examined in duplicate using the Formagraph

• (Foss Electric, Hiller~d, Denmark). Coagulation time (R), curd firming rate, and fmal curd

firmness (at 3xR) will be determined. Glucono-0-lactone will be used as the acidifying agent.

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This will give five levels of fat reduction and four different fat replacers. Eighty formagraph

tracings will be analyzed.

c. Determine the partitioning of fat replacers between curd and whey during coagulation and syneresis.

Small eight liter vats of Cheddar cheese will be manufactured from traditionally standardized

milk, 25% reduced fat, 50% reduced fat, 75% reduced fat, and non-fat milk with fat replacers

added. Whey will be collected and analyzed for the presence of micron-sized particles using the

Master Sizer instrument mentioned above. Entrapment of the hydrocolloidal fat replacing

particles in the curd will be examined by electron micrographs of the finished cheese samples.

d. Check heat and pH stability of fat replacers in the presence of milk proteins.

Differential scanning calorimetry will be used. to determine how heat influences solutions of

fat replacers without any milk protein. Then fat replacers and cream will be added to SkiJ!l milk

making 75% reduced fat, 50% reduced fat, and 25% reduced fat milks and heat treated in the

same manner. Skim milk and whole milk also will be analyzed. Electron micrographs of heat

treated skim milk with fat replacers will be taken to determine any differences before and after

heat treatment. Stability with changing pH will be studied by increasing and lowering the pH of

skim milk, 75% reduced fat, 50% reduced fat, 25% reduced fat and whole milk from pH 4.5 to

8.0 using increments of .5 pH units. Particles~ distribution and electron microscopy will be

used to check the pH stability of the hydrocolloidal particles and compared to milk fat

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Objective 2. Stability of fat replacers in dairy systems during growth of lactic starter cultures

a. Monitor growth and acid production rates of lactic acid bacteria in milk with fat replacers.

Strains of Lactococcus lactis ssp. lactis, Lactococcus lactis ssp. cremoris, Streptococcus

salivarius ssp. thermophilus, and Lactobacillus will be grown in milk and milk with each fat

replacer. Fat replacers and cream will be added to skim milk to make 75% reduced fat, 50%

reduced fat, and 25% reduced fat milk media. Skim milk and whole milk will be used as

controls. Growth curves at 21 °C and 32°C for mesophilic strains and 42°C for thermophilic

strains, using cell count and pH curves, will be done to determine the effect of each fat replacer

on the growth and acid production of each bacteria.

b. Measure the amount of enzymic (proteinase, peptidase and glucosidase) breakdown of milk proteins and fat replacers by lactic acid bacteria.

Proteinase-deficient and proteinase-positive strains of Lactococcus lactis ssp. lactis,

Lactococcus lac tis ssp. cremoris, Streptococcus salivarius ssp. thermophilus, and Lactobacillus

will be grown the M17, M17 with glucose, or MRS overnight at21°C and 32°C for mesophilic

strains and 42°C for thermophilic strains. Whole cells and filter-sterilized spent media will be

added.to buffer solutions at pH 6.8 and 5.2 containing Simplesse® to determine the prot~olytic

activity of strains toward Simplesse®. Fat replacers or cream will be added to skim milk and

different buffers to make 75% reduced fat, 50% reduced fat, and 25% reduced fat milk substrates

at pH 6.8 and 5.2. Whole cells and filter-sterilized spent media will be added. Proteolysis will

be determined using o-phthaldialdehyde and amino acid analysis as described by Weimer (1989) .

Lactic acid bacteria will be screened for exo- and endo-~-glucosidase activities using

p-nitrophenyl-~-D-glucoside (PNPG) and p-nitrophenyl-~-D-cellobioside (PNPC) as described

by Ohmiya et al. (1985). If results warrant further investigation, Stellarn.\ Paselli SA2 ®,and

Slendid® or cream will be added to skim milk to make 75% reduced fat, 50% reduced fat, and

25% reduced fat milk media. Degradation of the fat replacers in milk will be determined by

measuring the amount of glucose released using glucose oxidase colorimetric regent as described

by Ohmiya et al. (1985).

Ohmiya, K., M. Shirai, Y. Kurachi, and S. Shimizu. 1985. Isolation and properties of~glucosidase fromRhuminococcus a/bus. J. Bact. 161:432.

Weimer, B.C., C.J. Oberg, L.V. Moyes, R.J. Brown, and G.H. Richardson. 1989. Comparison of classical ion exchange amino acid analysis and o-phthaldialdehyde to characterize proteolysis by Lactobacillus bulgaricus. J. Dairy Sci. 72:2872 .

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c. Determine the manufacturing time and the amount of starter culture required to produce lowfat cheese containing fat replacers.

Strains of Lactococcus lac tis ssp. lactis, Lactococcus lactis ssp. cremoris, Streptococcus

salivarius ssp. thermophilus, and Lactobacillus will be grown in milk and milk with each fat

· replacer with chymosin and microbial rennets added. Fat replacers and cream will be added to

skim milk to make 75% reduced fat, 50% reduced fat, and 25% reduced fat milk media. Skim

milk and whole milk will be used as controls. Growth curves at 21 °C and 32°C for mesophilic

strains and 42°C for thermophilic strains, using cell count and pH, will be done to determine the

effect of each fat replacer on the growth and acid production of each bacteria.

The culture, milk coagulant, and fat replacer combination showing the favorable results will

be selected and trial vats of cheese will be made. Taste panels will be conducted to determine

consumer acceptability of the lowfat cheese.


Sales of "healthy" foods doubled between 1986 and 1990 and are expected to double

again by the year 1995 (The Cheese Reporter, 1991). Fat substitutes are predicted to be the

largest growth segment of the food ingredient industry in the 1990's (The Cheese Reporter,

1991). Retail sales of these "healthy" foods amounted to $14.3 billion in 1990, up sixteen

percent from 1989. Lowfat, low-cholesterol foods accounted for $8.2 billion of the total. Dairy

products accounted for $4.9 billion, mostly from frozen yogurt and dairy desserts.

The cheese market also has been expanding in recent years. Yet, lowfat cheese products,

particularly natural cheese, have been slow to appear in the market place. Development of more

lowfat cheese products requires an understanding of fat replacers and their interaction with milk

proteins. The important question is if fat replacers can mimic milk fat. By finding some answers

to this question, the dairy industry have more more scientific approach to using fat replacers in

lowfat dairy products than simply " depending more on the skill and art of the formulator than

the science of food technology" (Glicksman, 1991). This is the type ofresearch currently needed

by the dairy industry to be more involved in the the rapidly expanding, lowfat food market.

The Cheese Reporter. 1991. Sales of reduced-fat, other 'healthy' foods to double. October 18. Glicksman, M. 1991. Hydrocolloids and the search for the "oily grail". Food

Technology 45(10):94 .

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1. Proposed Title: Interactions between milk proteins and hydrocolloidal milk fat replacers

July 1, 1992 through June 31, 1994

Salaries Year 1 Year2 Total Graduate Student 10,000 10,500 20,500 Graduate Student 10,000 10,500 20,500

Research Scientist (25%) 10,000 10,500 20,500 Fringe Benefits 3,700 3,885 7,585

I It

Subtotal $33,700 $35,385 $69,085

Expenses Supplies 8,500 10,000 18,500

Travel 1,000 1,000 3,000 Publication Costs 1,000 2,000 3,000

Electron microscopy 3,000 3,000 6,000

TOTAL $47,200 $51,385 $99,585

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9. Summary of Proposed Project:

Heat treatment of milk is a technologically important process. This heat treatment brings about a series of protein-protein interactions bytween whey proteins and caseins in the milk. One such interaction between B-LG and K-CN has been identified in model systems and in milk. This interaction inhibits the rennet coagulation of milk, and is presumed to inhibit the age gelation of UHT milk and improve the physical properties of the yogurt gel. It ~s suggested that the dissociation of K-CN/B-LG complex from the surface of the micelles influences the age gelation of UHT milk, however, the dissociation of the complex during the storage of UHT milk has not been demonstrated and the exact mechanism of age gelation has not been worked out. Similarly, a positive correlation between the degree of whey protein denaturation and physical properties of yogurt gel (syneresis and firmness) has been shown , but no data exist on the quantitative relationship between the degree of K

CN/B-LG complex formation and properties of the yogurt gel. Recently, researchers at Utah State University have observed that the yogurt made (without any stabilizers) with UF concentrated skim milk with 5% protein heated at l20°C for 4 or 16 sec had superior textural properties (syneresis and firmness) compared to that heated at 82°C for 20 min or 130 or 140°C for sec. Although, it is generally accepted that whey proteins undergo denaturation at all these temperatures, no quantitative information on the degree of interaction between B-LG and K-CN at different temperatures which . might affect the properties of yoghurt is available.

Therefore, the research outlined in this proposal is aimed at (i) developing a quantitative relationship between the heating temperature and degree of complex formation between K-CN and BLG in both model systems as well as milk system, (ii) comparing the degree of complex formation between B-LG and K-CN in milk with the properties of yoghurt, (iii) measuring the dissociation of the complex in UHT milk during storage and compare with age gelation, and (iv) following the age gelation phenomenon in UHT milk by Raman Spectroscopy in order to understand the structural and conformational changes that occur in milk proteins during storage. The first part of the research will provide answers to some puzzling questions on the role of heat-induced interaction of B-LG with K

CN on the textural properties of yoghurt, so that, by manipulating the process variables a good quality yoghurt could be prepared without the addition of stabilizers. The second part of the research will enable us to understand the mechanism of age gelation in UHT milk, so that, appropriate measures could be taken to prolong its shelf-life. Thus, this research will address some fundamental issues concerning the molecular mechanisms that take place during the thermal processing of milk and relate these to the properties of end product made with such milks. This information would enable the dairy processor to produce better quality products, which will certainly ~~hanc.e the c<;>~su~~r appeal,_ r~sultin.g in the increased consumption of milk and milk products .

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• 4.1 Specific Objectives

Heat treatment of milk is a technologically important process. This heat treatment brings about a series of protein-protein interactions. between whey proteins and caseins in the milk ... One such interaction between B-LG and lC-CN has been identified in model systems and in milk. This interaction inhibits the rennet coagulation of milk, and is presumed to inhibit the age gelation of UHT milk. and improve the physical properties of the yogurt gel. It is suggested that the dissociation of lC-CN/B-LG complex from the surface of the micelles influences the age gelation of UHT milk, however, the dissociation of the complex during the storage of UHf milk has not been demonstrated and the exact mechanism of age gelation has not been worked out. Similarly, a positive correlation between the degree of whey protein denaturation and physical properties of yogurt gel (syneresis and firmness) has been shown , but no data exist on the quantitative relationship between the degree of lCCN/B-LG complex formation and properties of the yogurt gel. It has been shown that only the extent of interaction of B-LG with lC-CN affects rennet clotting properties of heated milk. Recently, researchers at Utah State University have observed that the yogurt made (without any stabilizers) with UF concentrated skim milk with 5% protein heated at 120°C for 4 or 16 sec had superior textural properties (syneresis and firmness) compared to that heated at 82°C for 20 min or 130 or 140°C for sec. Although, it is generally accepted that whey proteins undergo denaturation at all

these temperatures, no quantitative information on the degree of interaction between B-LG and lCCN at different temperatures which might affect the properties of yoghurt is available.

• Specific objectives and the hypothesis. The research outlined in this proposal is aimed at

•

(i) developing a quantitative relationship between the heating temperature and degree of complex formation between lC-CN and B-LG in both model systems as well as milk system, (ii) comparing

the degree of complex formation between B-LG and lC-CN in milk with the properties of yoghurt,

(iii) measuring the formation and dissociation of B-LG/K-CN complex in UHT milk during storage and compare with age gelation, and (iv) following the age gelation phenomenon in UHf milk by Raman Spectroscopy in order to understand the structural and conformational changes that occur in milk proteins during storage. The first part of the research will provide answers to some puzzling questions on the role of heat-induced interaction of B-LG with lC-CN on the textural properties of yoghurt, so that, by manipulating the process variables a good quality yoghurt could prepared.without the addition of stabilizers. The second part of the research will enable us to understand the mechanism of age gelation in UHT milk, so that, appropriate measures could be taken to prolong its shelf-life. Thus, this research will address some fundamental issues concerning the molecular mechanisms (eg., interaction of B-LG with K-CN) that take place during the thermal processing of milk and relate these to the properties of end product made with such milks .

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1. Proposed Title: Interaction Between Milk Proteins During Thermal Processing and its Effect on the Physicochemical Properties of Dairy Foods.


Salaries Year 1 Year2 Year3 Total Dr. Brown, Professor (5%) NoSal~ NoSal~ NoSal~ NoSal~ Dr. McMahon, Asst. Prof. (10%) NoSal~ No Salecy NoSal~ No Salary Dr. Reddx. Research Asst. Prof.(lOO%) 36,000 37,800 39,690 113,490 Fringe Benefits 13,320 14,175 15,083 42,578

Subtotal $49,320 $51,975 $54,773 $156,068 Expenses Year 1 Year2 Year3 Total Supplies 5,000 6,000 2,000 13,000 Travel 1,000 1,000 1,000 3,000 Publication Costsl 500 1,000 1,500 3,000 Other Expenses

UHT maintainance cost 6,000 6,000 Raman Spectroscopy (lease) 25,000 25,000

TOTAL $55,820 $65,975 $84,273 $206,068

1 Proposed journal(s ) to which the project results/manuscripts will be sent and the corresponding page charges: J. Dairy Sci., $70 per printed page .

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Heat treatment of milk is a technologically important process. This heat treatment brings about a series of protein-protein interactions between whey proteins and caseins in the mill<:. .. One such interaction between B-LG and 11:-CN has been identified in model systems and in milk. This interaction inhibits the rennet coagulation of milk, and is presumed to inhibit the age gelation of UHT milk.and improve the physical properties of the yogurt gel. It is suggested that the dissociationof K-CN/B-LG complex from the surface of the micelles influences the age gelation of UHT milk, however, the dissociation of the complex during the storage of UHT milk has not been demonstrated and the exact mechanism of age gelation has not been worked out. Similarly, a positive correlation between the degree of whey protein denaturation and physical properties of yogurt gel (syneresis and firmness) has been shown , but no data exist on the quantitative relationship between the degree of K-CN/B-LG complex formation and properties of the yogurt

gel. It has been shown that only the extent of interaction of B-LG with K-CN affects rennet clotting properties of heated milk. Recently, researchers at Utah State University have observed that the yogurt made (without any stabilizers) with UF concentrated skim milk with 5% protein heated at l20°C for 4 or 16 sec had superior textural properties (syneresis and frrmness) compared to that heated at 82°C for 20 min or 130 or 140°C for sec. Although, it is generally accepted that whey proteins undergo denaturation at all these temperatures, no quantitative information on the degree of interaction between B-LG and K-CN at different temperatures which might affect the properties of yoghurt is available.

Therefore, the research outlined in this proposal is aimed at (i) developing a quantitative relationship between the heating temperature and degree of complex formation between K-CN and B-LG in both model systems and in a milk system, (ii) comparing the degree of complex formation between B-LG and k-CN in milk with the properties of yoghurt, (iii) measuring the dissociation of the complex in UHT milk during storage and compare with age gelation, and (iv) following the age gelation phenomenon in UHT milk by Raman Spectroscopy . The first part of the research will provide answers to some puzzling questions on the role of heatinduced interaction of B-LG with k-CN on the textural properties of yoghurt, so that, by manipulating the process variables a good quality yoghurt could preparedwithout the addition of stabilizers. The second part of the research will enable us to understand the mechanism of age gelation in UHT milk, so that, appropriate measures could be taken to prolong its shelf-life. Thus, this research will address some fundamental issues concerning the molecular mechanisms that take place during the thermal processing of milk and relate these to the properties of end product made with such milks. This information would enable the dairy processor to produce better quality products, which will certainly enhance the consumer appeal, resulting in increased consumption of milk and milk products.

4.5 Available Facilities and Equipment

Standard laboratory facilities and equipment are available with in the Department of Nutrition of and Food Sciences at Utah State University. These include Sorval high speed centrifuges, Beckman and Shimadzu Spectrophotometers, Fourier Transform Infrared Spectrophotometer, F.P.L.C. and H.P.L.C. systems, Differential Scanning Calorimeter, Malvern Photon Correlation Spectroscopy system, Malvern Master Sizer, Delsa Coulter Counter, Phast System Electrophoresis, Beckman Amino Acid Analyser, Viscometers, Circulating water baths, pH meters, balances and three cold room laboratories. Fluorescence Spectrophotometer is available in Animal, Dairy and Veterinary Science Department on the campus and is accessible to us. Laser-Raman Spectroscopy to monitor conformational and secondary structural changes in UHT milk will be acquired for this project on a lease-basis or purchased .

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Project # 16. Effects of pH, Salts, and Thiolation ... Project #18. Interaction Between Milk Proteins ...

The TAC evaluation of proposal #16 ranked 6th of 14 proposals considered. The TAC evaluation of proposal #18 ranked 9th of 14 proposals considered. TheNDPRB suggests combining these two proposals with part of Dr. Bart Weimer's project "Interactions Between Milk Proteins, Starter Cultures and Hydrocolloidal Milk Fat Replacers." Specifically, the NDPRB suggests that objective #1 of Dr. Weimer's proposal be combined with your proposals #16 and #18.

The Director suggests that you consider combining your two proposals with Dr. Weimer's proposal. There is sufficient overlap among the various proposals' objectives that a more streamlined project can be written and resubmitted.

The Director suggests that budget revisions be made as follows:

Personnel

Dr. Reddy Graduate Student Fringe (37 .5 and 38.0%)

Expenses

Supplies , Travel Publication Costs Electron Microscopy Raman Spectroscopy (lease)

TOTAL

Year 1

28,000 12,000 10,500

10,000 1,000

500 3,000

$65,000

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Year2

29,400 12,600 11,172

12,000 1,000 1,000 3,000

25.000

$95,172

Total

57,400 24,600 21,672

22,000 2,000 1,500 6,000

25.000

$160,172

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Summary Recommendations for 1992-93 Project Approval

1 . Production of Extracellular Proteases of Brevibacterium linens for Use in Lowfat Cheese

Year 1: $35,625 Year 2: $39,293

2. Bacteriophage-Resistance Gene Replacement in Lactococcus lactis

$74,918

Year 1: $40,860 Year 2: $41,571 Year 3: $44,228 $126,659

3. Purification of Monospecific, Polyclonal Antibodies from Bovine Cheese Whey

Year 1: $44,875 Year 2: $42,360

4. Rheology and Microstructure of Mozzarella Cheese

To be submitted for linkage to NDPRB Competitive project

5. Function of Whey Proteins and Lactose in Age Gelation of UHT -Processed Milk Concentrate: Part 2

Year 1: $22,075 Year 2: $26,000

6. Extrusion Processing of Whey Proteins (Pending)

7. Effects of Iron Fortification on Chemical, Physical, Microbiological and Nutritional properties of Yogurt

Year 1: $26,700 Year 2: $29,000

8. Effects of pH, Salts, and Thiolation on Thermal Gelation of Caseins Alone and in Mixed Protein-Protein and ProteinPolysaccharide Gels W1.l.a

Interactions Between Milk Proteins, Starter Cultures, and Hydrocolloidal Milk Fat Replacers W1.l.a

Interaction Between Milk Proteins During Thermal Processing and Its Effects on the Physicochemical Properties of Dairy Foods

Year 1: $65,000 Year 2: $95,172

TOTAL $190,260 $273,396 $44,228

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$87,235

$48,075

$55,700

$160,172

$552,759

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!~ WESTERN DAIRY FOODS RESEARCH CENTER

Office of the Director Department of Nutrition and Food Sciences Utah State University Logan, UT 84322-8700 Telephone(801) 750-2102· FAX (801) 750-2379

\

MEMORANDUM

DA1E: May 29, 1992

TO: WDFRC Operational Advisory Committee Members WDFRC Principal Investigators

FROM: Paul A. Savello, Director

RE: Information about WDFRC Annual Meeting

The WDFRC Annual Meeting will be held July 9 and 10, 1992 on the campus of Oregon State University, Corvallis, Oregon. Floyd Bodyfelt has been busy making some arrangements for this upcoming meeting. OAC members and Pis/staff who will be attending this meeting should make their own travel arrangements and accornodation reservations for the stay in Corvallis. Four (4) hotels/motels are listed with some information about the room charges. Floyd states that the first two hotels on the list are now being recommended along with the previous two (numbers 3 and 4) which were used by many during the 1990 Annual Meeting.

1. Grand Manor Inn 935 N. W. Garfield Ph: 503-758-8571 Single room starting at $53.00

2. Motel Orleans )j 7 935 N.W. Garfield Ph: 503-758-9125 /rft' /( ftt ;__::rf Single room $32 to double $38 --------3. Nendels Inn 1550 N.W. 9th Ph: 503-753-9151 Single room $46 to double $55

4. Shanico Inn 1113 N.W. 9th Ph: 503-754-7474 or 1-800-432-1233 Single room $35 to double $42

The Annual Meeting will begin at 8:30a.m. on Thursday, July 9 in the Agriculture Production Room of the LaSells Stuart Center (26th and Western Boulevard). The phone number of this Center is 503-737-2402.

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WES1ERN DAIRY FOODS RESEARCH CENTER

OPERATIONAL ADVISORY COMMITIEE MEETING

August 10, 1991

MINUTES

The following are the minutes of the Operational Advisory Committee (OAC) Meeting of the Western Dairy Foods Research Center held on August 10, 1991 on the campus of Utah State University.

Committee members in attendance:

Janet Williams (NDPRB) Rodney Brown (USU) W. Lee Reese (out-going Utah Dairy Commission member) Sheldon Pratt (Oregon Dairy Produts Commission) Jeff Strandholm (Kraft General Foods, Inc.) Raj Narasimmon (Schreiber Foods, Inc.) Gale Moser (United Dairymen of Idaho) Douglas Wilrett (Marschall-Rhone Poulenc, Inc.) Boyd Gardner (in-coming Utah Dairy Commission member) Harry Papageorge (NDPRB Research Committee) Clint Warby (Utah Dairy Commission) Paul Savello (Director, WDFRC)

Visitors were also in attendance and asked to participate in discussions during the meeting.

I. Five (5) new project recommendations (from the Technical Advisory Committee meeting of June 3, 1991) were considered by the OAC. Actions were taken by the OAC on these recommendations as follows:

1. Project Title: Evaluation of iron-protein complexes in iron-fortified dairy products. PI: Arthur Mahoney (USU)

Motion: By Gale Moser. OAC direct Director Savello to accept further explanation on approved project and to act on his discretion accordingly provided budget presented is not exceeded.

Second: Boyd Gardner

Action: Passed.

2. Project Title: Identifying batch of origin of finished cheese made in continuous processes. PI: Lynn Ogden (BYU)

Motion: By Rodney Brown. Approve project providing matching industry funds and others being obtained and TAC recommendations (of June 3, 1991) being met.

Second: Lee Reese

Action: Passed

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3. Project Title: The influence of preadsorbed protein on adhesion of Listeria monocytogenes of dairy food contact surfaces . PI: Mark Daeschel (OSU)

Motion: By Floyd Bodyfelt. Approve contingent upon notification from NDPRB that project is not duplication of other work in similar research area.

Second: Sheldon Pratt

Action: Passed with one (1) dissenting vote

4. Project Title: Using whey for improvement of exposed subsoils and sodic and saline-sodic soils. PI: Conly Hansen (USU)

Motion: By Gale Moser. Approve project as submitted.

Second: Rodney Brown

Action: Passed

5. Project Title: Comparative effects of whey protein concentrate (WPC), lactose, salt, phosphate and pH on cooked yield, bind and acceptability of turkey rolls and boneless hams. PI: Daren Cornforth (USU)

Motion: By Rodney Brown. Approve project contingent upon recommendations ofTAC (June 3, 1991) being met at discretion of Director.

Second: Gale Moser

Action: Passed

II. One (1) recommendation (from Technical Advisory Committee meeting of June 3, 1991) to approve increased funding level for existing WDFRC project was considered. The action taken on this recommendation by the OAC was a follows:

1. Project Title: Prediction and determination of the efficacy of nisin in dairy foods. PI: Mark Daeschel (OSU)

Motion: By Rodney Brown. Approve project extension and additional funding as requested.

Second: Floyd Bodyfelt

Action: Passed

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III. The future focus and direction of the WDFRC was discussed. Director Savello presented slides describing the strengths of the WDFRC. Savello explained that the NDPRB is urging the Dairy Centers to carefully determine the focused research area(s) that each Center will pursue in the future (extension) years of Center activities. Input regarding the focused research area(s) of the WDFRC from OAC members (and from visitors) was solicited by Director.

Warby: The people (i.e. researchers/staff) at the WDFRC should be evaluated to help determine what the focused research area(s) should be.

Williams: The Center needs to know who will be doing what type of research.

C.A. Brostrom (visitor): The quality/experiences of the people in the Center detennine the Center's focus. The personnel of the WDFRC has changed since its inception in 1987. These changes must be considered in determining the Center's focused research area(s).

Brown: A strength of the WDFRC is its research activities in milk proteins and how these proteins apply to dairy products.

Williams: The Center should carefully select a "main focus" and augment this with ancillary /support research areas.

Wilrett: Focusing research can also be accomplished by making project proposals within the Center more directed.

Motion: By Clint Warby. To have the WDFRC consider the focused research area by "milk proteins" with research conducted in thermal processing, microbiology, cheese, and new opportunities for use of milk products and to have Director prepare revised proposal to be submitted to NDPRB and OAC with focused area (or 2 to 3 areas) of research.

Second: A second was presented.

Action: Passed

IV. The OAC set the dates July 9 and 10, 1992 for the 1992 Western Dairy Foods Research Center Annual Meeting, to be held in Corvallis, Oregon on the campus of Oregon State University .

1991-1992 Annual Report - DigitalCommons@USU

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