Effect of dietary palm oil fatty acids and buffer onphysiological performance of cows during the transition

period

Teama F. E. I., Osman S. F. and El-Tarabany A. A.,Department of Biological Applications, Radioisotopes Applications Division, Nuclear Research Center, Atomic EnergyAuthority, Cairo, Egypt, P.O.13759.

Key words: palm oil, buffer, transition period, cows

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1

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Abstract The main goal of the present study is to examine the physiological effect of supplemental palm oil fatty acids-ca salt and buffer on some biochemical and hormonal levels, milk yield and composition, for high yielding Holstein Friesian cows during transition period. To achieve this goal, thirty six (Holstein Friesian cows) aged 53±1.4 months and 739±19.86 kg body weight were used. The cows were randomly divided into three groups: 12 cow each, control group ( C ), the second group (T1) received 4% calcium salt palm oil fatty acid (Ca-POFA), the third group(T2) received Ca-POFA+ buffer (sodium bicarbonate) (0.5%) for three weeks before and after parturition (transition period). Blood samples were collected at 21, 14, 7 and 2 days before and 2, 7, 14 and 21 days after parturition to assess total protein, albumin, globulin, glucose, cholesterol, creatinine, total lipids and total cholesterol, in addition to cortisol and progesterone hormones. Milk yield were recorded for each animal and samples were taken afterparturition at different intervals (7, 14 and 21 day) to assess milk composition. A significant decrease of total protein, globulins, total lipids, cortisol and progesterone hormone levels in postpartum cows compared to pre-partum and this values were improved after (Ca-POFA) + buffer supplementation. On theother hand, increasing levels of serum albumin, cholesterol and creatinine were recorded in postpartum period which returned to the normal level as shown in albumin only after treatment with (Ca-POFA) + buffer. A continuous increasing of cholesterol and creatinine levels was observed as a result of supplement of (Ca-POFA) + buffer (sodium bicarbonate 0.5%). A significant, improvements in milk yield, 4% FCM and GE were determined after (Ca-POFA) + buffer supplementation. Whereas non significant changes were recorded in total solids, fat, protein and ash content of milk.

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In the present study, supplementation of cows in transition period with (Ca-POFA) + buffer (sodium bicarbonate 0.5%) can improved the level of some blood components and hormones as well as improve the milk yield which may reflects on the performance of dairy cows.

Introduction

Transition period is the most important step of life in the dairycow; it starts three weeks before calving and ends three weeks after calving. This period involves major changes in nutritional template, ethical and physiological patterns in cows. Performance and future health of dairy cows is based upon successful transfer of this period. Any disorders in this period may cause different diseases and a lot of economic losses (Drackley, 1999 and Badiey, 2009). It is the most critical period in a dairy cow's production cycle and lactation and any shortage of feed intake occurring during the final weeks of pregnancy whenthe nutrient demands for fetal growth and initiation of milk synthesis areincreasing result in higher energy requirements that can be met by energy dietary obtained from intake (Bell, 1995). A state of negative energy balance (NEB) has been occurred during the transition period (Grummer, 1995; Butler, 2001 and Kehrli et al., 2006), which characterized by dramatic changes in endocrine status and decrease in dry matter intake. These rapid changes allowing little time for the cow self recovery and influence metabolism and lead to mobilization of fat from adipose tissue and glycogen from the liver in addition to repair the cow for parturition and lactogenesis (NRC, 2001 and Grum et al., 1996). Fat feeding has become common practice in the dairy industry due to its ability to increase the energy content of the diet of lactatingdairy cows, therefore and hence, improve energy balance of dairy cows. In addition, inclusion of fat supplements into dairy rations reduces the incidence of metabolic disorders in dairy cattle (Grummer and Carroll, 1991) through positive effect on metabolic efficiency (Kronfeld et al., 1980). Feeding supplemental fat in the form of Ca salts of fatty acids (CSFA) makes the fat partially inert in the rumen and sometimes can prevent a depression in DMI. Feeding CSFA can allow specific fatty acids

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(FA) to escape ruminal biohydrogenation which are then available in the small intestine for absorption and utilization. Providing fat in the form of CSFA may reduce a deficiency and act on target tissues in addition to increasing the energy density of the diet ( Cullens, 2005). Feeding cows diet high in fermentable carbohydrates or lower ineffective fiber levels result in less chewing time and reduced production of saliva with its natural buffers. Ruminal pH varies considerably during the course of a day, and is particularly affected by the amount of fermentable carbohydrate in each meal. Shifts of 0.5 to 1.0 pH unit withina 24-hour period are common (Dado and Allen, 1993; Nocek et al., 2002). This represents a 5- to 10-fold change in hydrogen ion concentration in the rumen (pH is a log base 10 transformation of hydrogen ion concentration). Increasing the ruminal input of buffers from the diet or from saliva may help to prevent such a depression in ruminal pH. Shortage of ruminal buffers can be met by supplementing dietary buffers. Sodium bicarbonate is an important endogenous buffer of ruminal pH (Erdman, 1988)and it is also the most common buffer used in the dairy industry, sodium bicarbonate is produced naturally in cow's saliva but may be insufficient in rations of high acidity grass and maize silages and cereals. Sodium bicarbonate buffers maintain rumen pH at optimum level while improving rumen throughput. The goal of this study was to examine the physiological effects ofsupplemental palm oil fatty acids-Ca salt and buffer on some metabolites, some hormones and milk production or milk component for Frisian cows during transition period.

Materials and MethodsAnimals, Experimental Design and Rations: Thirty six Holstein multiparous, high yielding cows (900kg/season)at fifth parity aged (53 ±1.4) months with average body weight (739±19.86 kg). Cows were in the transition period (3 weeks before and after parturition). Animals owned by Modern Agriculture Investments Co., (Dina). Dina is located in a desert area at 80-km north on the western side of Cairo-Alexandria desert road. The farm exists at latitude 30° 30’N, longitude 30° 35’ E, and altitude 98 above sea level.

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Cows were housed in fan type open yard, and were semi-shaded with aluminum roof shade located in the middle of a coral with an area of 6m2 per cow under the shade and 18m2 per cow in the surrounding area. Duringpre and postpartum period, the cows were randomly divided into three equalgroups 12 cow each, control group (C) received 0% calcium salt palm oil fatty acids (Ca-POFA), group 2 (T1) received 4% (Ca-POFA) on the base of (DM), and group 3 (T2) received 4% (Ca-POFA)+ buffer (sodium bicarbonate 0.5% of TMR) purchased from (chichkulm, dis, tiarget, Turkey, imported by ElKiar for trading and engineering agencies, Alexandria, Egypt) for 3 weeks before and after parturition. The experimental period which extend from March to May. The cows were fed the same basal diet once a day with total mixed ration (TMR) formulated according to recommendations by NRC (2001) and Dina farms plan. Using (30kg/head/day),(TMR) composted of 12.50kg corn silage, 3.70 kg alfalfa hay(20 moisture), 6 kg crashed corn grains, 1.5kg soya bean meal, 1.2kg cotton seed cake, 2.5 flex seed cake, 2.60 kg glutofeed 25%.Chemical feed analysis during the experimental periods (Table 1).Palm Oil Fatty Acids calcium salt (Ca -POFA) composition: The composition of Ca-POFA is 78% brute fat, 3.5 % moisture, ashes12.5 (9% calcium), 6% methionin, 0% fiber. Fatty acids profile: 0.5% acid-insoluble ash, 1% myristic acid, 44% palmitic acid, 5% stearic acid, 40% oleic acid, 9.5 % linoleic acid. Energy value of (Ca-POFA) 33.25 MJ, (Magnapac is manufactured in Egypt by Sasam, Attaka Industrial Zone SuezGulf)Table (1): Chemical feed analysis during the experimental periods.Item Experimental periods

Prepartum PostpartumFeed analysis (%) ofDMDM 81 87CP 14 16.81NDF 52 42ADF 25 23.5CF 15.5 15.5NEI*(Meal/kg) 1.63 1.83

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KCaPMg

1.231.20.220.33

1.231.20.220.33

* NRC (2001).Blood samples and laboratory analysis: Fasting blood samples were taken at 21, 14, 7 and 2 day prepartumand 2, 7, 14 and 21 postpartum from the jugular vein using vacutainertubes centrifuged (1500rpm for 30 minutes) and sera were stored at -20 °Cuntil analysis. Serum samples were used to determine total protein,albumin, glucose, total cholesterol, total lipids and creatinine usingcommercial kits, purchased from (Bio-Diagnostic Company, Egypt).Cortisol and progesterone levels were determined by RIA technique usingsolid phase coated tubes kits purchased from Diagnostic SystemsLaboratories, (Inc., Webstir, Texas, USA). Collection of milk samples and measurement of milk composition Milk yield was recorded with electronic milk meters at Dina farm (Metatron P21, GEA Boenen, Germany) 3x daily 0600, 1400, and 2100. Approximately 300 mL of milk were obtained as proportional subsamples of total milk during day 7, 14, and 21. Milk yield data were stored electronically from all three groups. Total solids, fat, protein and ash contents of the milk were estimated according to Gupta et al. (1992). Solids-not-fat (SNF) content was calculated by subtracting fat content from total solids content, and lactose content was calculated by subtracting the sum of protein and ash contents from SNF content. Gross energy of milk was calculated by the following formula given by Tyrell andReid (1965): Gross energy of milk (kcal/kg) = 92.25F+49.15S-56.40 (Where, ‘F’ is Fat % and ‘S’ is SNF % in milk).Corrected level of fat, was calculated according to NRC (2001) as the following equation: 4% FCM = 0.4 x Milk + 15 x (Fat %/100) x Milk.

Statistical Analysis:

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Data were statistically analyzed using the General Linear ModelProcedure of GLM procedure by SPSS (SPSS version 8 for Windows; SPSS Inc.,Chicago, IL, USA). ANOVA model 2x3 was used to study the effect oftransition period (P), treatments (T) and the interactions between P andT. The model used is: Yij = ( +Pi + Tj + (PT)ij + eijk where (= theoverall mean; Pi = the fixed effect of the transition period (1=prepartum, and 2= postpartum ); Tj = the fixed effect of treatments (1=Ca-POFA and 2= Ca-POFA+buffer); (PT)ij=the interactions between the twofactors (P&T) and eijk = random error. Mlik data was analyzed using oneway ANOVA. Significance of the difference between the means was verifiedby Duncan’s new multiple ranges test (1955).

Results and Discussion1. Blood Metabolites

Total proteins, albumin and globulin concentrations were affectedsignificantly by the transition period , treatments and their interactions(p<0.001). The effect of two periods on mentioned metabolites showed that, the decrease in the percentages of change in total protein and globulin concentration in the postpartum as compared to pre-partum period by 16.74 and 41.84 %, respectively. While, the increase in the percentagesin albumin concentration was recorded in the postpartum as compared to pre-partum period by 18.45 % (Table 2).Our present results are in agreement with what observed by other authors in different studies. They attributed the low levels of plasma proteins observed during the first days after calving which to the shift of protein level in the last pregnancy period to face the fetal muscular development (El Sherif and Assad, 2001; Karapehlivan et al, 2007). The lower level of protein in postpartum cows may also due to the high energy need for milk synthesis inanimals during early lactation (Bremmer et al, 2000; Yokus et al, 2006). The improvements of treatments on the mentioned metabolites showed that, the increase of % of change in total protein concentration in cows treatedwith Ca- POFA and Ca- POFA + buffer as compared to control group by 4.98 and39.27 %, respectively. Also, the percentage of improvement was observed inglobulin concentrations in cows treated with Ca- POFA + buffer as compared

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to control group by 48.52 %, respectively. While, the increase in albumin concentration was recorded in cows treated with Ca- POFA and Ca- POFA + buffer as compared to control group by 23.6 and 29.2 %, respectively (Table 2). Previous data reported by Putnam and Varga (1998) exhibited the same observation and that improvement or increasing of protein levels may be attributed to the increase of nitrogen retention in late gestation in dairy cows. The authors believed that the rumen fermentation and microbial protein synthesis which favored by buffer may affect the protein levels in plasma specially in the prepartum phases where gastrointestinal tract fill is grater, turnover rate are slower, and ruminal retention times are longer (Hayirli et al. 2002). The interactions between transition periods and treatments showed that, the highest values of total protein and albumin were recorded in cows treated by Ca- POFA + buffer in postpartum period, while the lowest value was in controlgroup in the same period. Whereas, the highest value of globulin was observedin cows treated by Ca- POFA + buffer in pretpartum period, but, the lower valueof globulin was noted in cows treated by Ca- POFA in the postpartum period.Table 2: Influence of dietary Ca-POFA and buffer on total protein, albumin, globulin and glucose concentrations during pre- and postpartum ofthe transition period in dairy cows (M±SE).

ItemsBiochemical parameters

Total protein(g/dl)

Albumin(g/dl)

Globulin (g/dl) Glucose (mg/dl)

Periods (P)Prepartum 6.51A±0.01 2.71B±0.01 3.80A±0.008 53.47A±1.8Postpartum 5.42B±0.01 3.21A±0.01 2.21B±0.008 45.48B±1.8Significance 0.001 0.001 0.001 0.001Treatments(T)Control 5.22B±0.02 2.50B±0.01 2.72B±0.01 47.99C±2.2Ca- POFA 5.48B±0.02 3.09A±0.01 2.31C±0.01 49.71B±2.2Ca- POFA + buffer 7.27A±0.02 3.23A±0.01 4.04A±0.01 50.74A±2.2Significance 0.001 0.001 0.001 0.001P*T

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PrepartumControl 6.17ab±0.3 2.50d±0.02 3.67b±0.02 52.27 3.1±

Ca- POFA 6.16ab±0.3 2.76c±0.02 3.40c±0.02 53.81 3.0±

Ca- POFA + buffer 7.22a±0.4 2.76c±0.02 4.46a±0.02 54.35 3.2±PostpartumControl 4.28b±0.3 2.50d±0.02 1.78d±0.01 43.72 3.0±

Ca- POFA 4.65b±0.3 3.42b±0.02 1.23e±0.01 45.61 3.0±Ca- POFA + buffer 7.33a±0.4 3.70a±0.02 3.63b±0.01 47.13 3.2±Significance 0.001 0.001 0.001 0.29Means bearing different letters (A, B and C or a, b and c) in the same column within each classification differ significantly (P<0.001).Each figure represents the mean values of the samples collected at (21,14,7,2 and 2,7,14,21) during the transition period. Previous study was conducted by Grummer and Carroll, (1991) showed that concentrations of plasma glucose were not generally affected by supplementation of saturated or unsaturated fats (Moallem et al 2007). However, in this study, concentration of fasting serum glucose was affected significantly due to transition period and treatments (p<0.001). The effect of two periods on glucose showed that, the decrease in the level of postpartum as compared to pre-partum period by 14.94 %. While, the increase induced by treatments on glucose level was found in cows treated with Ca- POFA and Ca- POFA + buffer as compared to control group by 3.58 and 5.73 %, respectively. But interaction between periods and treatment (P*T) was not significant (Table 2). In a review article on dietary fat and adipose tissue metabolism, Chilliard (1993) noted that in52 comparisons study the difference in plasma glucose between control and fat-supplemented groups was (0 ± 0.03 g/L). The mammary glands have got high demand for glucose when itcoupled with insufficient carbohydrate supply causes blood levels ofglucose to drop and glucagon levels to increase. An increase in glucagon

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triggers lipase, which increases lipid breakdown for energy. When a cow isin negative energy balance resulting low insulin levels initiate themobilization of amino acids and lipid to compensate the decrease in energysupply for the cows (Fahey and Berger, 1988). Different influence ofPOFA application on metabolism of transion cows may result from thecontent of fatty acids in that supplements (McNamara et al. 2003). It wasshown that an application of rumen-protected fat in cows feeding causedthe decrease in glucose concentration in blood (Hammon et al. 2008) thisfinding came on contrary to our results. In the study made by Hammon et al.(2008) an increase in lactose yield after rumen-protected fat applicationwas observed. The authors concluded that the glucose sparing-effectallowed an enhanced lactose synthesis coincided with feeding rumen-protected fat. On the other hand, in the study Petit et al. (2004), anapplication of protected palm oil did not statistically influence theNEFA, glucose, cholesterol and HDL and LDL level. Using calcium salts oflong chain n-6 fatty acids (CaLFA), prilled fat and propylene glycol inthe transition period, higher yield and more profitable energy balance wasobserved in cows that were given supplements, as compared to control cows(Castañeda-Gutiérrez et al. 2009). Fat supplementation usually increases blood cholesterol (Garcia-Bojalil et al., 1998), but the type of fatty acid alters the effect. In this study there was a significant increase in total cholesterol concentrations during the postpartum period when compared to the prepartumperiod by 6.61 % (Table 3). The decrease in total cholesterol in late pregnancy has also been reported in cows and in other species: by Tainturier et al. (1984) in Friesan cows at prepartum period, Bekeová et al.(1987) before parturition Krajničáková et al. (2003) in goats. This is probably related to the role of the compound in ovary steroidgenesis, so that the total cholesterol concentrations are under the control of complexfactors. The administration of fed 4% POFA and 4% POFA + buffer caused a significant increases in serum cholesterol as compared with control groupsby 16.2 and 28.31%, respectively Also, the same result was observed in the effect of interaction between the period and treatment (P*T). These results did not agree with Tyagi, et al., (2010) who found that there

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were no difference in plasma cholesterol between the control and cows supplemented bypass fat. Previous studies have shown that postpartum serum cholesterol was in its lowest concentration at the first month after parturation, reachingits maximum concentration at about 5 months and decreased thereafter at late lactation. Both the increased nutrient demands of the rapidly growingfetus from the previous pregnancy and the utilization of cholesterol for steroidogenesis could be considered as contributing factors to a low levelin early lactation (Arave et al., 1975) which come in disagreement with the data of the present study.Table 3: Influence of dietary POFA and buffer on total cholesterol, total lipids and creatinine concentrations during pre- and postpartum of the transition period in dairy cows (M±SE).Items Cholesterol

(mg/dl)Lipids (mg/dl)

Creatinine (mg/dl) Periods (P)Prepartum 123.16B ±1.8 614.3A±19.4 1.56 B ±0.01Postpartum 131.30A ±1.8 576.0B±14.9 1.72 A ±0.01Significance 0.001 0.001 0.001Treatments (T)Control 110.80C±2.8 425.5C±18.2 1.06C±0.01Ca- POFA 128.75B±2.8 661.5B±18.2 1.90B±0.01Ca- POFA + buffer 142.17A±2.8 698.5A±18.2 2.02A±0.01Significance 0.001 0.001 0.001P*TPrepartumControl 113.0d±3.2 466.0e±26 0.96 0.02±Ca- POFA 118.1c±3.2 653.0d±26 1.74 0.02±Ca- POFA + buffer 138.3b±3.2 724.0a±26 2.00 0.02±Postpartum

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Control 108.6e±3.2 385.0f±25 1.05 0.02±Ca- POFA 139.3b±3.2 670.0c±25 2.06 0.02±Ca- POFA + buffer 146.0a±3.2 673.0b±25 2.05 0.02±Significance 0.01 0.01 0.31Means bearing different letters (A, B and C or a, b , c, d and f) in the same column within each classification differ significantly (P<0.01or 0.001).Each figure represents the mean values of the samples collected at (21,14,7,2 and 2,7,14,21) during the transition period. Total lipids concentrations decreased significantly by 6.23%, when compared prepartum period to the postpartum one (Table 3). Raphael et al. (1973) found that total lipids at 0 to 4 weeks postpartum averaged 292 mg/dl and increased to 508 mg/dl in weeks 5 to 10. Mc Namara et al. (1995)observed a minimum total lipid at parturition followed by an increase in postpartum and early lactation. This results disagree with data of the present study where total lipids decreased at postpartum than prepartum period. Milk fat production was positively correlated to plasma total lipids ( Hammon et al., (2009), they suggested that the cause was relatedto underfeeding in early lactation. Fatty acids of milk lipids (glycerides) arise from two distinct sources, from blood plasma lipids andfrom direct synthesis in the mammary gland. But there was a significant increase when the cows fed 4% POFA and 4% POFA + buffer by 55.46 and 64.15% respectively, and there is a significant interaction between the period and treatment (P*T). Fahey and Berger (1988) found that the high demand of energy for lactation and the mammary glands cause an increase inglucagon which triggers lipase, which increases lipid breakdown for energy. When, a cow is in negative energy balance the low insulin levels initiate the mobilization of amino acids and lipid to compensate the decrease in energy supply for the cows. Data of the present study shows a significant increase in totallipids in the treated group due to the feeding of palm oil fatty acids and buffer which may reduce the negative energy balance in postpartum cows. The elevated level of total lipids in late gestation is probably due

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to the reduced insulin-mediated inhibition of lipolysis observed in late pregnancy (Schlumbohm et al. 1997). Lipogenesis stimulated by insulin is also responsible for the increased values of total lipids observed in cowsduring early lactation. The significant post-partum increase in serum total lipids induces an increased FFA uptake by the liver from circulatingplasma, with increased triglyceride storage, as observed in cattle by Grummer (1993).

In the present study creatinine content of serum was statistically significant higher in postpartum than in prepartum period byabout 10.26%, with feeding on calcium salt fatty acids and buffer these increases were 79.25 and 90.6%, respectively and there is no interaction between the period and treatment. The quantity of creatinine formed each day depends on the total body content of creatine, which in turn depends on dietary intake, rate of synthesis of creatine, and muscle mass. It was shown that in dairy cows, plasma creatinine and muscle diameter decreased through the period from week 1 before calving to week 4 after calving. Moorby et al (2002) this is disagree with ore data.

2. Hormonal levels: Cortisol and progesterone levels were higher during the prepartum, and then dropped rapidly during the postpartum phases as shown in table (4) where it was 82.45% for cortisol and 88.24%for progesterone as compared to prepartum levels. This decrease was significant as affected by this period. These changes may explain their main role in lactogenesis in cows (Dann and Varga, 1999). The decrease in cortisol level in cows treated with Ca- POFA and Ca- POFA + buffer as compared to control group by 5.47 and 6.56 %, respectively was significant also, interaction between the period and treatment was significant. Cortisol is the major stress hormone produced by the adrenal glands in the ruminant. During the transition period, cortisol iselevated prior to, during and following parturition, indicating an increased release of the trophic hormone, adrenocorticotropin

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(ACTH) (Hunter et al., 1970). For many years glucocorticoids havebeen used in the medicine for their immunosuppressive effects. Because cortisol peaks are found at the time of parturition, the dairy cow’s immune system is compromised. Progesterone is the product of the corpus luteum and is delivered to the uterus via circulation. It is the essential steroid hormone for maintenance of pregnancy. From day 32 to 256 of gestation period plasma progesterone concentrations remained fairly constant, with a significant reduction just prior to parturition (Short 1958). Plasma progesterone levels in the pregnant dairy cow increases steadily until about day 250 ofgestation, when the peak concentrations becomes between 7 and 8 ng/ml. After ward, until the day of calving, the level drop to be between 3 to 4ng/ml while, plasma estrogens are concomitantly increasing. The last day prepartum, progesterone drops down to almost undetectable levels. Both progesterone and estrogen levels remain low from calving until about 3 weeks postpartum. The postpartum increase in progesterone, determined by aconcentration greater than 1 ng/ml, was not detected until about day 23 postpartum (Eley et al., 1981). Data of the present study showed a sharp drop of progesterone level by 88.24% in postpartum as compared to prepartum level and this decrease was significant (P<0.001) which confirmsthe results of the pervious studies.Table 4: Influence of dietary Ca-POFA and buffer on hormonal levels duringpre-and postpartum (the transition period) in dairy cows (M±SE).

Items Hormonal levels Cortisol (ng/ml) Progesterone (ng/ml)

Periods (P)Prepartum 18.63 A ±0.11

5.10A±0.02

Postpartum 3.27 B ±0.120.60B±0.02

Significance 0.001 0.001Treatments (T)Control 12.80 A ±0.13

3.23A±0.03

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Ca- POFA 12.10B±0.13 2.65B±0.03Ca- POFA + buffer 11.96B±0.13 2.67B±0.03Significance 0.001 0.001P*TPrepartumControl 20.8a±0.19 5.9a+0.04Ca- POFA 17.6b±0.19 4.7b+0.04Ca- POFA + buffer 17.5b±0.19 4.7b+0.04PostpartumControl 4.8c±0.18 0.56c±0.05Ca- POFA 2.6d±0.18 0.61c±0.05Ca- POFA + buffer 2.4d±0.18 0.65c±0.05Significance 0.05 0.001Means bearing different letters (A, B and C or a, b, c and d) in the same column within each classification differ significantly (P<0.05 or 0.001). Each figure represents the meanvalues of the samples collected at (21,14,7,2 and 2,7,14,21) during the transition period.

It was noticed that dietary fats typically increase concentrations of circulating cholesterol, the precursor of progesterone (Grummer and Carroll, 1991). Ruminants fed supplemental fat often have a slight increase in blood progesterone concentration (Staples et al., 1998). This finding disagree with our present results where during postpartum period, there is a significant decrease in progesterone level between control group and (T1) and (T2) by about 17.96% and 17.34% respectively, and a highly significant interaction between period and treatment (P*T). Hawkins et al. (1995); Staples and Thatcher, (2005) suggested that the increase seen in circulating progesterone when cows were fed supplemental fat was from a reduced rate of progesterone clearance rather than from an increased synthesis of progesterone. It was also shown that cows supplemented with rumen inert polyunsaturated fat hadgreater mean serum progesterone concentrations compared with control (Lopes et al., 2009; Salas-Razo et al., 2011). Collectively, the hormonallevels for both cortisol and progesterone were decreased in postpartum period compared to prepartum one and supplements of 4% POFA + buffer

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improved the decreasing of their levels in the postpartum period. The improvement of hormonal levels obtained in the present study after treatment may be attributed to the improvement in rumen fermentation process as well as microbial protein synthesis due to presence of buffer added during the treatment. 3. Milk yield, 4% FCM: Table (5) depicted that response to supplemental Ca-POFA, for average milk yield,4% fat corrected milk (FCM) were significantly higher (p<0.001) in group that were fed 4% Ca-POFA+ buffer (T2) (17.6 kg) milk yield than in control group by (3.53%) and by about 2.33% than the group of (T1) which fed Ca-POFA only. 4% FCM was increased by 5.26% in group (T2) than control group and 4.1 than the group of (T1) respectively. Gross energy (GE) was significantly higher (p<0.01) in cows treated by Ca-POFA + buffer as compared with control group by 1.4%. These results are similar to the data obtain by Shaver (1990) who observed an increase of daily milk yield when cows fed supplemental fat. King et al. (1990) He also reported that a supplement of predominantly saturated and unesterified long chain fatty acids in the ration of cows increased the yield of milk and 4% FCM by 3.3 and 6.3 kg /cow/day, respectively. Grummer(1995) indicated that average of fat-corrected milk response to supplementation with dry granular fats (average of supplementation 2.3 percent of DM) vs. tallow or vegetable oils (average supplementation 2.65 percent of DM) when diets already contained whole oilseeds was 1.1 vs. 0.1kg/d, respectively. Calcium salts of palm oil fatty acids (Ca-POFA) are now firmly established worldwide as the standard source for dairy energy supplements (Onetti and Grummer, 2004). An increase in milk production has also been reported by Palmquist (1984) and Sarwar et al. (2003) in cows supplemented with rumen-protected fat up to 6%. Milk-yield response to supplemental fat can be influenced by several factors, including basal diet, stage of lactation, energy balance, fat composition, and amount of supplemental fat. Kronfeld (1976) indicated that milk production reaches its maximum efficiency when FAs constitute 16 percent of metabolizable energy.

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The little improve in milk yield associated with supplemental fatcan largely be attributed to an improvement in energy balance (Grummer andCarrol, 1991). Maeng et al. (1993) also reported a significant (p<0.05) increase in milk production in cows supplemented with calcium salt of longchain fatty acids which corporate with the findings of the present study. On contrary to the present finding Johnson et al. (1988), Casper et al. (1990) and Elliot et al. (1993) reported that, no effect of supplemental fat on 4% FCM yield. Also, Han, et al., (2011) found that supplementationof palm oil by-pass fat for early lactating Holstein cows, the dry matter intake (DMI) and milk yield were not affected, however, the milk protein, milk fat, no-fat solid, and total solid increased numerically. This may bedue to that during the first two weeks, the cows did not acclimated to theration completely.Table (5): Mean ± SE. of milk yield and compositions of dairy cows as affected by palm oil fatty acids and buffer during postpartum period (21 day).

Parameters Experimental groups Significancevalues Control (C) Ca-POFA Ca-POFA+ buffer

Milkyield(kg)

17.0C ± 1.3 17.2B ± 1.1 17.6 A±1.2 0.0014% FCM(kg) 15.2B±1.4 15.4B ±1.2 16.0A±1.4 0.001G E (kcal/kg) 660.9C±12.4 655.9B±6.2 670.1A±7.2 0.01Milk compositions (%): Total solids 11.7 11.6 11.8 0.19Fat 3.3 3.3 3.4 0.23Proteins 3.5 3.5 3.5 0.21Lactose 4.22A 4.14B 4.22A 0. 01Ash 0.68 0.68 0.68 0.43SNF 8.40 8.30 8.40 0.18Means bearing different letters (A, B and C) in the same row within each classificationdiffer significantly (p<0.01or 0.001).

4. Milk composition: Table (5) showed that there was a significant decrease in lactose level by 1.9 % in group fed Ca-POFA (T1) than in cows in group (T2) fed Ca-POFA + buffer and control group. Hammon et al. (2008) observed an

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increase in lactose yield after rumen-protected fat application. In general milk composition was not affected due to supplementation of Ca-POFA in the ration of postpartum cows. Schneider et al. (1988), Jerred et al. (1990) and Klusmeyer et al. (1991) reported that milk protein percentage was not changed by addition of calcium salts of long chain fatty acids. which corporate to the findings of the present study.Similarly, Brzoska et al. (1999) reported that increasing amount ofcalcium salt of palm oil fatty acids had no negative effect on milk fat, protein and lactose contents of milk Of 83 treatment comparisons (fat supplementation vs. control) summarized by Wu and Huber (1994), milk protein production was unchanged or increased in 65 comparisons and decreased in 26. However, in 15 of the 26 comparisons in which protein production was decreased, milk production also was decreased. On contrary to present results, Sarwar et al. (2003) observed an increase in milk fatand protein percentage in buffalos as a result of fat supplementation in ration Solomon et al. (2000) observed an increased milk fat percentage incows with supplemental fat in the diets. On the other hand Palmquist, (1984) and Grummer and Carrol (1991) observed a reduction in milk fat percentage when unprotected tallow was fed to dairy cows which was attributed to reduced fiber digestion and altered acetate: propionate ratio. The increase in protein percentage in milk due to addition of inertfat might be due to reduction of usage amino acid as fuel and thus spared for casein synthesis (Sarwar et al., 2003). Therefore, the milk composition was not affected due to supplementation of palm oil calcium salt plus buffer.

Conclusion The present data indicated that supplementation of cows in transition period with (Ca-POFA) + buffer can improve the level of some blood components and hormones as well as improve the milk yield but not its composition which may reflect on the performance of dairy cows. More studies are required to evaluate the relationship between many levels of Ca-POFA or buffer and animals' productivity under several environmental conditions.

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