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Biological Trace Element Research ISSN 0163-4984Volume 155Number 1 Biol Trace Elem Res (2013) 155:29-37DOI 10.1007/s12011-013-9728-2

Assessment of Chromium Content ofFeedstuffs, Their Estimated Requirement,and Effects of Dietary ChromiumSupplementation on Nutrient Utilization,Growth Performance, and Mineral Balancein Summer-Exposed Buffalo Calves(Bubalus bubalis)Muneendra Kumar, Harjit Kaur, AmrishTyagi, Veena Mani, Rijusmita SarmaDeka, et al.

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Assessment of Chromium Content of Feedstuffs, Their EstimatedRequirement, and Effects of Dietary Chromium Supplementationon Nutrient Utilization, Growth Performance, and Mineral Balancein Summer-Exposed Buffalo Calves (Bubalus bubalis)

Muneendra Kumar & Harjit Kaur & Amrish Tyagi &Veena Mani & Rijusmita Sarma Deka & Gulab Chandra &

Vijay Kumar Sharma

Received: 15 May 2013 /Accepted: 4 June 2013 /Published online: 21 August 2013# Springer Science+Business Media New York 2013

Abstract This study was conducted to determine the chro-mium content of different feedstuffs, their estimated require-ment, and effect of dietary Cr supplementation on nutrientintake, nutrient utilization, growth performance, and mineralbalance in buffalo calves during summer season. Levels ofCr was higher in cultivated fodder, moderate in cakes andcereal grains, while straw, grasses, and non-conventionalfeeds were poor in Cr content. To test the effect of Crsupplementation in buffalo calves, 0, 0.5, 1.0, and 1.5 ppmof inorganic Cr were fed to 24 buffalo calves. Buffalo calveswere randomly assigned to four treatments (n=6) and raisedfor 120 days. A metabolic trial for a period of 7 days wasconducted after 3 months of dietary treatments. Blood sam-ples were collected at fortnight interval for plasma mineralestimation. The results suggested that dietary Cr supplemen-tation in summer did not have any affects (P>0.05) on feedconsumption, growth performance, nitrogen balance, andphysiological variables. However, dietary Cr supplementa-tion had significant effect (P<0.05) on balance and plasmaCr (ppb) levels without affecting (P>0.05) balance andplasma levels of other trace minerals. The estimated Crrequirement of buffalo calves during summer season wascalculated to be 0.044 mg/kg body mass and 10.37 ppmper day. In conclusion, dietary Cr supplementation has re-gardless effect on feed consumption, mass gain, and nutrient

utilization in buffalo calves reared under heat stress condi-tions. However, supplementation of Cr had positive effect onits balance and plasma concentration without interactingwith other trace minerals.

Keywords Chromium . Buffalo calves . Growthperformance . Nutrient intake . Nitrogen balance .

Mineral balance

Introduction

The effects of heat stress are costly to dairy farmers due toreduced performance and animal production. Increased en-vironmental temperature elevates the respiratory rate andwater intake [1]; reduces the gut motility, rumination, andruminal contractions [2]; and depresses appetite [3], by hav-ing a negative effect on appetite center of the hypothalamus[4]. Buffaloes exhibit signs of great physical distress whenexposed to environmental stress as compared to other farmanimals [5]. This is due to fact that their body absorbs a greatdeal of solar radiation because of their dark skin and sparsecoat or hair. In addition to that, they possess a less efficientevaporative cooling system due to their rather poor sweatingability. There are opportunities to recover the losses due tohot weather by modifying environment, genetic develop-ment of breeds that are less sensitive to heat and nutritionalmanagement [6]. In domestic animals, Cr has been recog-nized as a newer essential trace mineral [1, 7] and suggestedto alleviate stress-associated effects [8–10]. Supplementationof Cr has been shown to improve performance of growingand lactating ruminants exposed to thermal stress [11].Considering these facts, the present study was conducted in

M. Kumar (*)Animal Nutrition Department, College of Veterinary Scienceand Animal Husbandry, DUVASU, Mathura, Indiae-mail: muneendra82@gmail.com

H. Kaur :A. Tyagi :V. Mani : R. S. Deka :G. Chandra :V. K. SharmaDairy Cattle Nutrition Division, National DairyResearch Institute, Karnal, India

Biol Trace Elem Res (2013) 155:29–37DOI 10.1007/s12011-013-9728-2

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buffalo calves to determine the effect of inorganic chromiumsupplementation on the nutrient utilization, performance,and nitrogen and mineral balance during summer season.

Materials and Methods

Selection of Animals and Their Management

The experiment was conducted in the cattle yard of NationalDairy Research Institute (NDRI), Karnal, Haryana. Karnal issituated on an altitude of 250 m above mean sea level,latitude and longitude position being 29° 42″ N and 79°54″ E, respectively. The climate of the Karnal is arid tosemiarid with average rainfall of 455 mm and average tem-perature 7 to 39 °C.

Prior to the commencement of the experiment, all proto-cols were reviewed and approved by the Institutional AnimalEthics Committee constituted as per the article number 13 ofthe committee for the purpose of control and supervision onexperiments on animals rules, laid down by the Governmentof India. In present study, 24 female buffalo (Bubalus buba-lis) calves (10 to 13 months and 180±5 kg body mass) wereselected from the herd of NDRI, Karnal, and randomlyassigned to four treatment groups (n=6), on body mass andage basis. The experimental animals were housed in separateconcrete floor pens. The duration of the experiment lasted120 days. Deworming of all the animals was done before thestart of the experiment and experimental animals were letloose every week for exercise. Experimental animals eitherreceived a basal diet devoid of supplemental Cr (control) orwere supplemented with 0.5, 1.0, and 1.5 ppm trivalent Crfrom an inorganic source (CrCl3·6H2O, molecular weight266.45, minimum assay 97 %; Hi-Media Laboratories Ltd,Mumbai, India). The nutrient requirements of calves weremet by feeding concentrate mixture and maize or jowarfodder [12]. To ensure that each calves consumed Cr, theCrCl3 was mixed with 50 g concentrate mixture, and scat-tered onto a small amount of green fodder before offering thediet. The experimental animals were daily monitored for drymatter intake (DMI) and physiological variables. Calveswere weighed and blood samples were collected at 0, 15,30, 45, 60, 75, 90, 105, and 120 days of Cr supplementation.

Metabolic Trials

A metabolic trial for a period of 7 days was conducted after3 months of dietary treatments. All the buffalo calves werehoused in individual shaded metabolic stalls with their re-spective dietary treatments and fresh drinking water wasprovided thrice a day. The diet was weighed daily and thenoffered to each animal with 4-day adaptation period followedby a 7-day collection period. Daily feeds offered and residue

left was recorded to determine the net feed intake. Feces andurine voided during 24 h was recorded for 7 days and dailyrepresentative samples were taken for estimation of chemicalcomposition, nitrogen, and mineral balance.

Proximate Analysis and Fiber Fraction

Samples of feed, residue, feces, and urine were analyzed forDM, organic matter (OM), total ash, ether extract, nitrogen(N) [13], and neutral detergent fiber (NDF) and acid deter-gent fiber (ADF) [14].

Trace Elements Estimation

For Cr estimation, collected samples of feeds and feces wereoven dried (at 80 °C for 16 h) and ground to pass through a 1-mm sieve. The ground samples (5 g) were placed in glazedceramic crucibles and ignited in a muffle furnace at 450 °C for4 h. The ash was treated with concentrated nitric acid under mildheat to oxidize the trivalent Cr (Cr3+) to hexavalent Cr (Cr6+) formore accurate detection. The acid-extracted cooled sampleswere filtered through ash less Whatman filter paper no 1. Thecrucibles were washed several times with deionized water andthe final volume was made up to 10 ml for the estimation of Cr.A solution of ammonium chloride (20 g/l) was added in thestandard and the samples to reduce interferences due to thepresence of Fe in the samples. Concentration of Cr in feed wasestimated in the atomic absorption spectrophotometer (Model Z-5000, Polarized Zeeman Atomic absorption Spectrophotometer,Hitachi High-Technologies Corporation, Tokyo, Japan).

Concentration of zinc (Zn), copper (Cu), iron (Fe), andmanganese (Mn) in feed, feces, urine, and plasmawas estimatedin the atomic absorption spectrophotometer (AAS, Model Z-5000, Polarized Zeeman Atomic absorption Spectrophotometer,Hitachi High-Technologies Corporation, Tokyo, Japan) afterdigestion in tri acid mixture (HNO3/HClO4/H2SO4− 2.5:1:1ratio). The procedure described in AAS manual (1988) forpreparation of stock and standard solutions and choice of instru-mental conditions were followed.

Climatic Data Recording

Microclimatic data, viz., dry bulb temperature (Cdb) andwet bulb temperature (Cwb) in degree Celsius, wererecorded at 7.30 A.M. and 2.30 P.M. with dry and wet bulbhygrometer (Zeal, UK) every day during experimental pe-riod using the equation (temperature humidity index(THI)=0.72×(Cdb+Cwb)+40.6) [15]. Daily relative humid-ity (RH) was calculated by difference of dry bulb temper-ature and wet bulb temperature.

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Physiological Variables

Respiration rate (RR) of the animals was recorded by ob-serving flank movement and one inward and outward move-ment was counted as one respiration and recorded/minute.To record the pulse rate (PR), coccygeal artery was palpatedand recorded/minute. Immediately after recording pulse rate,rectal temperature (RT) was recorded using a mercury inglass clinical thermometer (Qingdao Dacon Trading Co.Ltd., Shandong, China), inserted 7 cm in the rectum for atleast 2 min and the rectal mucosa was in contact with thebulb of the thermometer.

Blood Sampling and Biochemical Analysis

Blood samples were collected in sterile heparinized vacu-tainer (Becton Drive, Franklin Lakes, NJ, USA) tubes fromjugular vein puncture, posing minimum disturbance to theanimal on days 0, 15, 30, 45, 60, 75, 90, 105, and 120 post-supplementation of Cr. Day “0” represented the start ofexperiment. Immediately after collection, blood sampleswere centrifuged at 1,200×g for 20 min to separate theplasma for the analysis of trace mineral.

Statistical Analysis

Data were subjected to analysis of variance using the generallinear model procedure of the Statistical Software Package(SPSS for Windows, V19.0; SPSS Inc., Chicago, IL, USA)according to the following model:

Y ijk ¼ μþ Ti þ Dj þ T � Dð Þij þ eijk

Where

Yijk Dependent variableμ Overall mean of the populationTi Mean effect of the treatmentDj Mean effect of day of sampling (j=0, 15, 30, 45,

60, 75, 90, 105, and 120 days of dietary treatment)(T×D)ij Effect of the interaction between effect of Cr

supplementation and day of samplingeijk Unexplained residual element assumed to be

independent and normally distributed.

Individual animals were used as the experimental unit forall data. The pairwise comparison of means was carried outusing “Tukey’s honestly significant difference (HSD) test.”Significance was determined at P<0.05 and the values arepresented in the tables. Multiple regression analysis wasused to calculate the requirement of chromium element.

Y ¼ aþ b1X 1 þ b2X 2

Where

a ConstantY Average element intake (milligram)X1 Average body mass (kilogram)X2 Average element balance (milligram)b1 Element intake per kilogram body massb2 Element balance per unit element intake.

Equation:

Y summerð Þ ¼ aþ b1X 1 þ b2 X 2

¼ 0:254þ 0:012 x 232:81þ 14:96 x 0:487¼ 10:37mg=day ppmð Þ¼ 0:044 mg=kg bodymass

Results

Chromium in Feedstuffs

Table 1 summarizes the Cr content of different feedstuffs.Levels of Cr were found higher in cultivated fodder samples;however, cakes and cereal grains aremoderate source of Cr.Mostof the straw, grasses, and unconventional feeds tested containedless than 1.0 ppm Cr and considered poor source of Cr.

Climatic and Physiological Variables

During experimental period, THI and RH% was increasingfrom May and reached maximum in July (Fig 1). But meanTHI value throughout the experimental period of 120 dayswas 80.30±0.57. Mean value of THI recorded during exper-imental period indicated that calves were in moderate heatstress.

No significant impact (P>0.05) of Cr treatments wasdetected on the RR, PR, and RT across 120 days summerseason trial (Table 4).

The proximate composition, fiber fraction and mineralcontent of the basal diet are presented in Tables 2 and 3.The diet had adequate crude protein (CP) and total digestiblenutrients (TDN) to support maintenance and growth of buf-falo calves [12]. The total Cr content of the basal dietsformulated was determined to be 0.23 mg/kg DM, whereasconcentrate mixture, maize, and jowar fodder contains 0.22,0.28, and 0.19 mg Cr/kg DM, respectively.

Nutrient intake in term of DM, CP, digestible crude pro-tein (DCP), and TDN was not affected (P>0.05) by supple-mentation of inorganic Cr (Table 5). Similar to nutrientintake, digestibility coefficients of nutrients was also notaffected (P>0.05) by Cr supplementation (Table 6). Thetrend of performance expressed as body mass change during120 days in summer season was also found similar in all fourrespective groups, indicating that Cr supplementation did nothave any effect (P>0.05) on growth performance (Table 4).

Effect of Chromium on Growth Performance in Summer-Exposed Buffalo Calves 31

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Nitrogen Balance, Mineral Balance, and Plasma MineralStatus

Table 4 shows the effects of Cr supplementation on the Nbalance in buffalo calves fed with Cr. Results of present studyindicated that N balance in buffalo calves fed with Cr weresimilar (P>0.05) to non-Cr-fed buffalo calves and all buffalocalves were found to be in positive nitrogen balance. In thepresent study, Cr balance increased with increased (P<0.05)dietary Cr level (Table 5). Similarly, fecal and urinary excretionof Cr (milligram per day) was also increased with increase inthe level of supplemental Cr. Retention of Cr was significant

different (P<0.05) in four respective groups and found maxi-mum in 1.5 ppm Cr-fed calves.

No effect (P>0.05) of Cr supplementation up to 1.5 ppmin the diet of buffalo calves was evident on the utilization ofFe, Cu, and Zn. Intake, outgo, and retention of Fe, Cu, andZn were found similar in all four respective groups. Changein plasma Cr concentration in the present finding evidencedthat there was significant (P<0.05) effect of dietary Crsupplementation (Table 6). Plasma Cr level was found sig-nificant higher in 1.5 ppm-fed buffalo calves. However,plasma concentration of Cu, Zn, and Fe was found similar(P>0.05) in all four respective groups.

Table 1 Cr content (in parts per million) of different feedstuffs

Sample no. Feedstuffs Botanical name Cr content (ppm)

Roughage

1. Barley fodder Hordeum vulgare 1.25

2. Berseem fodder Trifolium alexandrinum 1.31

3. Jowar fodder Sorghum bicolor 1.00

4. Maize fodder Zea mays 1.72

5. Oat fodder Avena sativa 1.37

6. Doob grass Cynodon dactylon 0.65

7. Guinea grass Megathyrsus maximus 0.22

8. Wheat straw Triticum aestivum 0.37

9. Rice straw Oryza sativa 0.22

Grain and grain by-products

10. Barley grain Hordeum vulgare 0.75

11. Wheat grain Triticum aestivum 1.02

12. Maize grain Zea mays 0.82

13. Rice bran Oryza sativa 0.22

14. Wheat bran Triticum aestivum 0.65

15. Guar korma Cyamopsis tetragonoloba 0.97

Cake

16. Cotton seed cake Gossypium sp. 0.70

17. Ground nut cake (expeller extracted) Arachis hypogaea 0.90

18. Ground nut cake (solvent extracted) Arachis hypogaea 0.72

19. Mustard oil cake (expeller extracted) Brassica campestris 0.50

20. Mustard oil cake (solvent extracted) Brassica campestris 1.02

21. Soyabean meal Glycine max 1.10

Non-conventional feed resources

22. Babul pods Acacia nilotica 0.22

23. Vilayati babul Prosopis juliflora 0.25

24. Subabool leaf Leucaena leucocephala 0.62

25. Bakeria Platanthera bakeria 0.47

26. Ker Pogonia ophioglossoides 0.35

27. Kheemp Leptadenia pyrotechnica 0.62

28. Kheijri Prosopis cineraria 0.22

29. Mural Licum berberum 0.42

30. Punarava Boerhavia sp. 0.15

31. Sataveri Asparagus racemosus 0.40

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Estimated Cr Requirement

Table 6 represents Cr intake, outgo, and balances in buffalocalve during summer season in four respective groups. Crintake during summerwas 5.12, 9.08, 12.09, and 15.22mg/day,respectively. The multiple regression equation showed highcorrelation (R2=96.55 %) between the different variables se-lected (Table 7). The estimated requirement of Cr was calcu-lated to be 0.044 mg/kg body mass and 10.37 ppm per dayduring summer.

Discussion

Cr Content in Feedstuffs, Climatic, and PhysiologicalVariables

Cr content in different feed stuffs collected from differentlocations is in consistent with findings of other workers whoalso found higher Cr content in fodder than other feeds [16].Leaves have higher Cr content than other plant parts whichmight be the reason for the higher Cr content in fodders [17,18]. The information regarding Cr concentrations in feed-stuffs is scanty so the results cannot be discussed on that way.

* Fortnight

THI -Temperature humidity index

RH- relative humidity

Fig. 1 Temperature humidityindex and relative humiditychanges during experimentalperiod

Table 2 Ingredient and nutrient compositions of basal diet

Ingredients Content (% as DM)

Berseem fodder 19.8

Corn silage 14.3

Ground yellow maize 28.9

Groundnut cake 15.7

De-oiled mustard cake 5.2

Wheat bran 6.4

Rice bran 8.3

Dicalcium phosphate 0.5

Salt 0.3

Trace minerals and vitamins premixa 0.6

CrCl3·6H2Ob Variablec

a Premix composition per kilogram: vitamin A, 500,000 IU; vitamin D3,10,000 IU; vitamin E, 100 mg; Ca, 190,000; P, 90,000; Na, 50,000; Cu,300 mg; Fe, 3,000 mg; Mn, 2,000 mg; I, 100 mg; Co, 100 mg; Se, 1 mg;Mg, 19,000 mg; BHT antioxidant, 3,000 mgb CrCl3·6H2O (minimum assay 97 %; Hi-Media Laboratories Ltd,Mumbai, India)c Supplemental Cr diets (0.5, 1.0, and 1.5 ppm Cr) were obtained bysupplementing Cr with 0.5, 1.0, and 1.5 mg/kg dry matter in basal diet

Table 3 Chemical compositions of basal diet

Attributes Content (% as DM)

Dry matter, % 75.6

Organic matter, % 78.3

Crude proteins, % 17.5

Crude fiber, % 24.0

Total ash, % 9.5

NDF, % 38.6

ADF, % 25.9

Calcium, % 1.00

Phosphorus, % 0.41

Magnesium, % 0.23

Sodium, % 0.20

Chloride, % 0.41

Copper, mg/kg 23.22

Zinc, mg/kg 49.54

Manganese, mg/kg 63.83

Iron, mg/kg 304.6

Chromium, mg/kg 0.23

Effect of Chromium on Growth Performance in Summer-Exposed Buffalo Calves 33

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Analysis of Cr in feedstuffs is also a challenging task due tothe low levels of Cr and problems with Cr contamination offeed samples during collection and preparation of samplesfor analysis [19].

THI observed during trial period is considered as moder-ate stress. However, other researchers stated 25 °C ambienttemperature as the upper critical temperature and 72 as theupper critical THI [20–22]. In the present study, Cr supple-mentation at an inclusion rate 0.5, 1.0, and 1.5 mg/kg frominorganic Cr could not alter physiological variable. Findings

of present study on physiological variables are consistent withprevious finding with inorganic sources of Cr supplementationin transportation-stressed calves [23] who also did not reportany difference in PR, RR, and RT. In contrast to the presentfindings, supplementation of 1.0 ppm Cr in calves exposed to42 °C ambient temperature reported to have a lower RR, PR,and RT [24]. No effect of supplemental Cr on RR, PR, and RTin the present studymight be due to the fact that the calves werenot exposed to such a high ambient temperature which couldresult in changes in these physiological parameters.

Table 4 Effect of Cr supplementation on physiological variables, growth performance and nutrient intake

Parameters Supplemental Cr (mg/kg of DMI, ppm) SEM P value

0 0.5 1.0 1.5

No. of buffalo calves 6 6 6 6

Days in trial 120 120 120 120

Rectal temperature, °C 38.19 38.14 38.21 38.14 0.13 0.998

Pulse rate, min−1 57.68 59.62 60.52 58.78 0.48 0.886

Respiration rate, min−1 45.60 46.18 44.84 16.09 0.75 0.666

Initial body mass, kg 183.79 180.86 183.41 185.77 19.74 1.000

Final body mass, kg 236.72 233.38 236.21 239.77 19.92 1.000

Gain, kg 52.93 52.52 52.80 54.00 0.23 1.000

Daily gain (kg/day) 0.441 0.437 0.440 0.450 0.20 0.999

DM intake (kg/day) 5.25 5.13 5.23 5.31 0.37 1.000

CP intake (kg/day) 0.650 0.646 0.648 0.656 0.27 0.998

DCP intake (kg/day) 0.422 0.416 0.421 0.426 0.21 1.000

TDN intake (kg/day) 3.48 3.43 3.46 3.50 0.23 1.000

Gain/DM intake ratio 0.084 0.077 0.075 0.078 0.001 0.998

Dietary composition, nutrient intake, digestibility, and performance

Table 5 Effect of Cr supplementation on nutrient digestibility and nitrogen utilization

Parameters Supplemental Cr (mg/kg of DMI, ppm) SEM P value

0 0.5 1.0 1.5

Digestibility coefficients (%)

DM 69.59 69.81 69.45 68.23 2.49 1.000

OM 71.84 73.62 72.88 73.17 2.51 0.956

CP 65.50 65.76 65.17 65.72 2.17 0.999

EE 77.37 79.39 78.39 76.19 3.95 0.895

NDF 64.27 70.31 68.28 69.32 2.02 0.855

ADF 46.67 44.19 46.48 46.65 1.68 0.800

Nitrogen balance

N intake, g/day 107.31 104.33 106.05 108.22 2.7 0.788

Fecal N excretion, g/day 34.06 33.21 35.35 34.69 0.9 0.999

Urinary N excretion, g/day 31.17 27.71 29.85 29.19 1.63 1.000

N retention, g/day 42.09 43.41 40.85 44.34 1.60 0.888

N retention , % of absorbed N 39.22 41.60 38.51 40.97 1.81 0.999

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Nutrient intake and digestibility in Cr-fed buffalo calves isin agreement with findings of other worker who alsoreported similar nutrient digestibility in calves supplementedwith 0.2 and 0.4 ppm Cr as CrCl3 [25–28]. Similar to thepresent study, supplementation of 0.37 ppm Cr picolinate(CrP) in the diet of Holstein calve [29] and 0.5 ppm in lambs[30] did not improve the intake of DM. Holstein calvessupplemented with 1.0 ppm Cr from high Cr yeast have14 % high DM intake [31]. No information is available inthe literature on the effect of supplemental Cr on the perfor-mance of buffalo calves during summer season. But incalves, supplementation of 0.75 ppm inorganic Cr do nothave any effect on growth performance [26] and which wassimilar to findings of present study. No effect of supplement-ing chelated Cr or chromium nicotinate (CrNic) on growthperformance of transportation-stressed calves was observed

by many authors [31–35]. Similarly, no difference in averagedaily gain, DMI, and gain/feed ratio in calves followingdietary supplementation of 1.0 ppm Cr was also observedby other researchers [36–38]. In contrast to the present find-ings, beneficial effect on performance of calves with supple-mental trivalent Cr as Cr yeast, chelated Cr, or CrNic duringthe stress period have been recorded by various workers[33–35]. A possible explanation could be that pooled Cr inthe body of these buffalo calves was not depleted as a resultof high ambient temperature. Another possible explanation isthat the good conditioning of buffalo calves allowed them tobetter tolerate the stressors of summer.

Supplemental Cr as CrP did not influence any measure ofN balance [39]. However, our results are in contrast withthose who stated that supplementation of 37 μg Cr as CrCl3significantly increased (P<0.05) N retention in lambs [40].

Table 6 Effect of Cr supplementation on trace mineral utilization and plasma trace mineral level

Parameters Supplemental Cr (mg/kg of DMI, ppm) SEM P value

0 0.5 1.0 1.5

Cr balance

Cr intake, mg/day 5.12 9.08 12.09 15.22 0.87 0.001

Fecal Cr excretion, mg/day 4.61 8.22 10.99 13.63 0.78 <0.001

Urinary (mg/day) 0.32 0.48 0.57 0.74 0.09 0.040

Cr retention, mg/day 0.19 0.38 0.53 0.85 0.06 0.001

Cr retention, % of absorbed Cr 3.68 4.22 4.36 5.61 0.10 <0.001

Plasma Cr level, ppb 0.19 0.31 0.40 0.44 0.08 0.010

Cu balance

Cu Intake (mg/day) 113.07 111.60 112.83 114.54 9.70 0.999

Cu voided in feces and urine, mg/day 97.88 96.59 97.67 99.15 6.15 1.000

Cu retention, % of absorbed Cu 13.44 13.45 13.43 13.44 0.02 1.000

Plasma Cu level, ppm 0.72 0.69 0.79 0.79 0.07 0.868

Zn balance

Zn intake, mg/day 1,424.44 1,405.90 1,421.40 1,442.97 122.23 0.868

Zn voided in feces and urine, mg/day 1,067.50 1,053.00 1,065.52 1,081.56 92.08 0.999

Zn retention, % of absorbed Zn 25.08 25.12 25.05 25.06 0.09 0.999

Plasma Zn level, ppm 1.31 1.31 1.32 1.33 0.11 1.000

Fe balance

Fe intake (mg/day) 3,282.15 3,239.43 3,275.15 3,324.84 281.64 1.000

Fe outgo in feces and urine, mg/day 2,422.76 2,391.07 2,417.68 2,454.32 208.02 1.000

Fe retention, % of absorbed Fe 26.19 26.19 26.18 26.18 0.05 1.000

Plasma Fe level, ppm 1.97 1.95 1.88 2.02 0.05 0.865

Table 7 Multiple regressionanalysis of Cr requirement inbuffalo calves during summerseason

Cr intake (mg) (Y) Body mass (kg) (X1) Cr retained (mg) (X2) Constant (a)

Summer season (R2=96.55 %, N=24)

Coefficient 0.012 14.96 0.25

Average 10.37 232.81 0.48

Effect of Chromium on Growth Performance in Summer-Exposed Buffalo Calves 35

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This apparent discrepancy might be attributed to the chemicalform of the Cr used (i.e., organic CrP or Cr yeast vs. inorganicCrCl3) or differences in the Cr status of the studied animals.

Similar to present study, increased Cr intake, excretion,and retention was also reported by other worker in stress-exposed animals [41]. In opposite to findings of the presentstudy, CrCl3-supplemented heifer as compared to control andCr yeast-supplemented heifer have high intake of Zn, Cu, Fe,and Mn [25]. Previous studies indicated that the apparentabsorption coefficient of Cr from inorganic trivalent sourcesranged between 4 and 10 % whereas 10 to 25 % for naturallyoccurring sources such as brewer’s yeast [42]. Whereas,dose-dependent linear decrease in the apparent absorptionof Zn was observed in CrCl3-fed heifers [43]. Blood Crconcentration might reflect to a certain extent the intake ofthis element, but in cases of excessive Cr intake, it is inap-propriate to use the blood Cr concentration as an indicator ofCr status in animals [42]. Dose-dependent linear increase intissue Cr concentration was observed in slaughter studies andfound blood Cr level of 0.125, 0.197, 0.225, and 0.240 μg/l,respectively in control, 0.5, 1.0, and 1.5 mg dietary Cr-supplemented groups [44]. Findings of the present experi-ment indicated that supplementation of Cr did not affect(P<0.05) plasma concentration of Cu, Zn, and Fe. Similarto present finding, supplementation of 2 mg of Cr/kg of dietas high Cr yeast in steers did not affect tissue trace metalconcentrations [32]. Similarly, rats fed with Cr at 5 mg/kg ofdiet did not have any change in plasma concentration of Cu,Zn, and Fe [45].

Conclusion

Straw, grasses, and unconventional feeds tested in presentstudy were found less in Cr content and feeding of these feedstuffs does not met the Cr requirement specially duringsummer months. The results of the current study indicatedthat dietary supplementation of Cr, in the form of chloride,does not have beneficial impacts on physiological variables,nutrient intake, nutrient utilization, and growth performancein buffalo calves under heat stress conditions. Additionally,dietary Cr supplementation influences the plasma Cr levelwithout affecting balance and plasma levels of other traceminerals. The estimated Cr requirement of buffalo was cal-culated to be 0.044 mg/kg body mass and 10.37 ppm per dayduring summer. Altogether, findings suggested that supple-mental Cr level is not sufficient to enhance performance andto ameliorate adverse effect in buffalo calves during summermonths.

Acknowledgments I would like to express my gratitude to all thosewho gave me the possibility to complete this research work. I want tospecially thank Dr. Harjit Kaur, for his great efforts of supervising and

leading us, to accomplish this fine work. I am thankful to the NationalDairy Research Institute, Karnal, for the institutional fellowship awardfor carrying out this research work.

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