Screening and characterization of Sahiwal cattle calves-origin lactic acid bacteria based on desired probiotic attributes for potential application.

Probiotics are living microorganisms that confer health benefits to host when administered in adequate amounts. To develop novel host-specific probiotic for their application as feed additive, the present study was undertaken to isolate and characterize probiotic strains of indigenous cattle-calves origin. A total of 55 colonies were isolated from 12 healthy calves, with 34 of the isolates being Gram-positive, catalase-negative and vancomycin-resistant. Furthermore, eleven isolates showed tolerance to acid (pH 2.0) and thirteen isolates tolerated bile salts (0.3%). Seven common acid and bile tolerance strains were further investigated for other probiotic attributes and displayed higher (p< 0.05) auto-aggregation and cell surface hydrophobicity values. Moreover, all seven isolates had potent antibacterial activity against pathobiont E. coli as well as significant co-aggregation capacity and enzyme activity. In vitro biosafety assessment revealed that all seven isolates were non-hemolytic, negative for mucin degradation and susceptible to most of the antibiotics. Based on the obtained findings, heatmap and principal component analysis identified four highly effective probiotic candidates confirmed by 16S rDNA sequencing as Limosilactobacillus reuteri SW23, Limosilactobacillus reuteri SW26, Limosilactobacillus reuteri SW27 and Enterococcus faecium SW28, respectively. Further studies on biosafety aspect are warranted for the application of these strains in animal as potential probiotics.HIGHLIGHTSL. reuteri SW23, L. reuteri SW26, L. reuteri SW28 and Enterococcus faecium SW28 were successfully isolated and identified from indigenous calves' feces.These microbes were characterized for potential probiotics attributes.Heatmap analysis and principal component analysis (PCA) was used along with probiotic attributes to select highly effective probiotic candidates.

Rumen microbial diversity, enteric methane emission and nutrient utilization of crossbred Karan-Fries cattle (Bos taurus) and Murrah buffalo (Bubalus bubalis) consuming varied roughage concentrate ratio.

Dietary mix and host species have both been shown to have a significant impact on rumen microbial diversity, enteric methane emission and animal performance. The goal of this study was to see how the roughage concentrate ratio 70:30 (Low concentrate; LC) vs 40:60 (High concentrate; HC) and the host species crossbred cattle vs buffalo affected rumen microbial diversity, enteric methane emissions and nutrient utilization. Dry matter intake (kg/d) and dry matter percent digestibility were considerably (p < 0.05) higher in the HC ration and buffalo compared to LC ration and crossbred cattle, respectively. Both dietary mix and host species had a substantial (p < 0.05) impact on intake of various nutrients, including organic matter (OM), crude protein (CP), ether extract (EE), neutral detergent fiber (NDF), and acid detergent fiber (ADF). Increased concentrate proportion in the ration improved nitrogen balance, resulting in increased average daily gain and considerably reduced methane (g/d) output (p < 0.05). Furthermore, 16S rRNA genes were sequenced using Oxford Nanopore Technology (ONT) and subsequently annotated using the Centrifuge workflow to uncover ruminal bacterial diversity. Firmicutes was considerably (p < 0.01) greater in the LC diet, whereas, Bacteroidetes was higher in the HC ration. Genus Prevotella dominated all rumen samples, and buffalo fed LC ration had significantly (p < 0.01) higher Oscillospira abundance. At the species level, simple sugar-utilizing bacteria such as Prevotella spp. and Selenomonas ruminantium predominated in the crossbred cattle, but fibrolytic bacteria such as Oscillospira guilliermondii were statistically (p < 0.01) more abundant in the buffalo. Overall, dietary mix and host species have both been shown to have a significant impact on rumen microbial diversity, enteric methane emission and animal performance, however, host species remained a major driving force to change ruminal community composition as compared to roughage concentrate ratio under similar environmental conditions.

Dietary supplementation of Aloe vera extract modulates rumen microbes and improves the functional food value of milk by altering phenolic content, antioxidant capacity, and fatty acid profile in lactating goats.

Twenty-four cross-breed (Alpine × Beetal) lactating goats (average body weight: 37.5 ± 2.7 kg and milk yield: 1.78 ± 0.31 kg/day) were chosen for the ninety days duration of the experiment and divided into three groups following a completely randomized design. Group I acted as control (T0) and received only a basal diet as per requirement, whereas group II (T1) and group III (T2) received basal diet added with Aloe vera extract at 2% and 4% of dry matter intake (DMI), respectively. Total phenolic compounds (TPC) and milk antioxidant status were higher (p < 0.05) in T1 and T2 than T0. Relative abundance of methanogen, protozoa, Butyrivibrio proteoclasticus, and Ruminococcus flavefaciens in the rumen were lower, while Butyrivibrio fibrisolvens population was higher (p < 0.05) in T2 and T1 compared to T0. Saturated fatty acids levels in milk were lower, whereas different polyunsaturated fatty acids levels were higher (p < 0.05) in T1 and T2 than T0. Nutritional indices of milk increased in both supplemented groups. Overall, it may be concluded that dietary supplementation of Aloe vera extract at 2% and 4% of DMI may enhance the functional characteristics of milk by boosting TPC and antioxidant status, as well as the FA profile and nutritional quality indices of milk fat.

Host-specific probiotics feeding influence growth, gut microbiota, and fecal biomarkers in buffalo calves.

The current study is aimed to evaluate the effect of host-specific probiotics on the gut microbiome, performance, and select fecal biomarkers of gut health in preruminant buffalo calves. Eight Murrah buffalo calves (3-5 days old; 32.52 ± 0.43 kg average body weight (BW)) were randomly allocated into two groups as follows; 1) Group I (n = 4) fed basal diet alone (CON); 2) Group II (n = 4) supplemented with a lyophilized probiotic formulation at a dose rate of 1 g/day/head (1 × 109 CFU/g) having Limosilactobacillus reuteri BF-E7 and Ligilactobacillus salivarius BF-17 along with basal diet (PF) for 30 days. Results revealed that final BW (kg), average daily gain (g/day), average dry matter intake (g/day), and structural growth measurements were significantly (P < 0.05) increased in the probiotics supplemented group (PF) compared to the control (CON). Fecal pH, fecal moisture, and fecal score were reduced (P < 0.05) in PF than in CON. Moreover, levels of fecal propionate, lactate, and ammonia altered positively in PF compared with CON. The relative abundance of Firmicutes tended to be higher (P = 0.10) in the probiotics fed group than CON. However, the relative abundance of Proteobacteria was significantly lower (P = 0.03) for calves fed probiotics on day 15. A trend was observed in Bacteroides (P = 0.07) and Lactobacillus (P = 0.08) abundances in the feces of the PF than in CON. Overall, it can be concluded that the administration of probiotic formulations significantly improved the performance and gut health of buffalo calves via modulating the gut microbiota composition.

Autochthonous Limosilactobacillus reuteri BFE7 and Ligilactobacillus salivarius BF17 probiotics consortium supplementation improves performance, immunity, and selected gut health indices in Murrah buffalo calves.

Probiotics have emerged as biotherapeutic adjuncts to combat neonatal calf gastrointestinal disorders. Therefore, they are considered a suitable alternative to antibiotics for maintaining a healthy and balanced gut microbiota. Hence, the current investigation was carried out to evaluate the effect of autochthonous probiotics on Murrah buffalo calves. Sixteen calves (5-7 days of age) were randomly divided into four groups. Group I served as control (CT), fed a basal diet with no supplementation. Groups II (LR), III (LS), and IV (CS) were supplemented with Limosilactobacillus reuteri BF-E7, Ligilactobacillus salivarius BF-17, and a consortium of both probiotic strains at a rate of 1x108 CFU/g/calf per day along with the basal diet, respectively. Two previously isolated potential probiotic strains, Limosilactobacillus reuteri BF-E7 and Ligilactobacillus salivarius BF-17, were found to be compatible in vitro. Dietary supplementation of probiotics for sixty days significantly increased (P<0.05) dry matter intake (DMI, g/d), average daily gain (ADG, g/d), net body weight gain (kg), feed conversion efficiency (FCE), and structural growth measurements as compared to control. Furthermore, a considerable (P<0.05) increase in the abundance of beneficial intestinal microbiota (lactobacilli and bifidobacteria) was observed along with improvement in fecal biomarkers like lactate and ammonia, immune status, and reduced fecal score. Upon comparative analysis among treatment groups, the results were found to be better in the probiotic consortium fed group compared to the LR and LS treated groups. The present findings conclusively deduced that autochthonous probiotic consortium might serve as potential candidate for fostering performance, immunity, and gut health biomarkers in Murrah buffalo calves.

Supplementation of Aloe vera extract in lactating goats' diet: effects on rumen fermentation efficiency, nutrient utilization, lactation performance, and antioxidant status.

The present work was conducted to investigate the effects of supplementing Aloe vera extract on rumen fermentation efficiency, nutrient utilization, lactation performance, and antioxidant status of goats. Twenty-four crossbreed lactating goats (Alpine × Beetal) were divided into three experimental groups (AV0, AV2, and AV4). AV0 had no supplementation, groups AV2 and AV4 received ready to feed aqueous extract of Aloe vera at 20 and 40 g/kg dry matter intake, respectively, along with basal diet and experiment lasted for 100 days. Average DMI did not vary (P > 0.05) among treatment groups; however, the metabolic bodyweight of AV4 was significantly lower (P < 0.05) than the AV0 and AV2 groups (AV0 = AV2 > AV4). Intake and digestibility of DM, OM, CP, NDF, ADF, and EE were unaffected (P > 0.05) by Aloe vera supplementation. The milk production, yield of milk fat, protein, lactose, and solid not fat (SNF) of goats in the AV4 group were significantly higher (P < 0.05) than other groups (AV4 > AV2 = AV0). The activity of superoxide dismutase and catalase enzymes and levels of plasma ferric reducing total antioxidant power were high (P < 0.01) in the Aloe vera supplemented group (AV4 = AV2 > AV0). There was no significant difference (P = 0.979) in the pH, acetic acid (P = 0.449), and butyric acid (P = 0.864) concentration of the rumen liquor among the treatment groups. The propionic acid concentration was similar between AV2 and AV4 and significantly higher (P = 0.024) than the AV0 group (AV4 = AV2 > AV0). Moreover, C2:C3 values were significantly lower (P = 0.037) in the AV4 group compared to the control (AV0). Thus, Aloe vera supplementation enhanced milk yield, propionic acid production, and antioxidant status without affecting nutrient utilization; however, results were better in the AV4 group. The inclusion of Aloe vera at 40 g/kg of DMI would improve the rumen fermentation efficiency, lactation performance, and overall health status of the dairy goats.

Optimizing fiber and protein levels in diet of lactating Murrah buffaloes to ameliorate heat stress: Effect on physiological status and production performance.

The objective of study was to assess the outcome of feeding six total mixed rations (TMR), differing in NDF and protein content, for their synergistic effect on ameliorating heat load of lactating Murrah buffaloes evident through improved physiological and production performance. Thirty six lactating Murrah buffaloes (587 ± 12.3, MY 9 ± 2.2, Parity 2.5 ± 1.5) were arranged in a 3 × 2 factorial design with three levels of dietary NDF (30, 34.5 and 37% dietary NDF) and two levels of metabolizable protein (MP; 7.0% and 8.4%). Buffaloes were fed either of six dietary treatments: 30%NDF; 7.0% MP (CF1, as recommended), 34.5%NDF; 7.0% MP (MF1), 37%NDF; 7.0% MP (HF1), 30%NDF; 8.4% MP (CF2), 34.5%NDF; 8.4% MP (MF2) and 37%NDF; 8.4% MP (HF2). TMR offered with maize silage and respective concentrate for 90 days feeding trial. Fortnightly feed samples and weekly milk samples collection was done for analyses. Metabolic trial conducted in mid of experiment for estimating nutrient digestibility. Throughout the trial, THI level (79.7-83.8) denoted that buffaloes were exposed to stressful environment. Higher MP in diet reduced pulse rate in buffaloes as compared with lower MP diet. Rectal temperature was lower in Murrah buffaloes fed MF2 diet whereas; minimum breathing rate was recorded for high protein fed group. The MF2 diet increased dry matter intake (kg/d) by 2.7%, milk yield (kg/d) by 8.3% and feed efficiency (milk/DMI) by 7.2% as compared with CF1 group indicating reduced heat load. Increase in protein intake along with improved protein digestibility in MF2 group was recorded. Measured 6%FCM and ECM (kg/d), milk fat (%) and total solid (%) were higher in MF2 treatment group. Results revealed that 34.5% NDF and 8.4% MP have a positive influence on amelioration of heat stress in present experimental conditions.

Effect of Milk Fermented with Lactobacillus acidophilus NCDC15 on Nutrient Digestibility, Faecal Biomarkers and Immune Response in Murrah calves

Abstract In neonates, rapid change in diet imbalances gut health allowing colonization of opportunistic pathogens that confer harmful effects on animal health causing reduced digestion and malabsorption of nutrients. In this milieu, probiotic feeding can be a promising approach in promoting animal health and stabilization of gastrointestinal microbiota. Hence, the present study was designed to investigate the effect of Lactobacillus acidophilus NCDC15 enriched fermented milk on nutrient digestibility, faecal biomarkers and immune response in Murrah buffalo calves. Twenty-four, neonatal calves (5-7 days) were randomly allocated into four groups for 90 days. The control group (CT) was provided a basal diet of calf starter and green fodder (maize and jowar), without any probiotic fermented milk (PFM) supplementation. Basal diet was supplemented with probiotic fermented milk at 100, 200 and 300 mL/calf/day, in PFM100, PFM200 and PFM300 groups, respectively. Nutrient digestibility remained similar among the groups. Faecal acetate was higher (P<0.05) in PFM200 and PFM300, while, faecal butyrate was increased (P<0.05) with all levels of probiotic supplementation than control. Faecal Lactobacillus and Bifidobacterium count were increased (P<0.05) with a concomitant reduction in coliform population (P<0.05) among all the treatments. Cell-mediated and humoral immune response were higher (P<0.001) in PFM200 and PFM300 than CT. Overall, it can be concluded that inclusion of Lactobacillus acidophilus NCDC15 in the form of fermented milk upto 300 mL/calf/day improved immunity and faecal biomarkers in Murrah buffalo calves without any adverse effect on nutrient utilization which may positively impact growth performance in Murrah buffalo calves.

Conjugated linoleic acid producing potential of lactobacilli isolated from goat (AXB) rumen fluid samples.

OBJECTIVE: The present investigation was aimed to explore the potential of lactobacilli for conjugated linoleic acid (CLA) production, isolated from rumen fluid samples of lactating goats. METHODS: A total of 64 isolates of lactobacilli were obtained using deMan-Rogosa-Sharpe (MRS) agar from rumen fluid of goats and further subjected to morphological and biochemical characterizations. Isolates found as gram-positive, catalase negative rods were presumptively identified as Lactobacillus species and further confirmed by genus specific polymerase chain reaction (PCR). The phylogenetic tree was constructed from the nucleotide sequences using MEGA6. RESULTS: Out of the 64 isolates, 23 isolates were observed positive for CLA production by linoleate isomerase gene-based amplification and quantitatively by UV-spectrophotometric assay for the conversion of linoleic acid to CLA as well as gas chromatography-based assay. In all Lactobacillus species cis9, trans11 isomer was observed as the most predominant CLA isomer. These positive isolates were identified by 16S rRNA gene-based PCR sequencing and identified to be different species of L. ingluviei (2), L.salivarius (2), L. curvatus (15), and L. sakei (4). CONCLUSION: The findings of the present study concluded that lactic acid bacteria isolated from ruminal fluid samples of goat have the potential to produce bioactive CLA and may be applied as a direct fed microbial to enhance the nutraceutical value of animal food products.

Presence of Acetamide in Milk and Beef from Cattle Consuming AFEX-Treated Crop Residues.

AFEX treatment of crop residues can greatly increase their nutrient availability for ruminants. This study investigated the concentration of acetamide, an ammoniation byproduct, in AFEX-treated crop residues and in milk and meat from ruminants fed these residues. Acetamide concentrations in four AFEX-treated cereal crop residues were comparable and reproducible (4-7 mg/g dry matter). A transient acetamide peak in milk was detected following introduction of AFEX-treated residues to the diet, but an alternative regimen showed the peak can be effectively mitigated. Milk acetamide concentration following this transition was 6 and 10 ppm for cattle and buffalo, respectively, but also decreased over time for cattle while tending to decrease (p = 0.08) for buffalo. There was no difference in acetamide concentration in the meat of cattle consuming AFEX-treated residues for 160 days compared to controls. Further investigation is necessary to determine the metabolism of acetamide in ruminants and a maximum acceptable daily intake for humans.

Isolation, characterization and conjugated linoleic acid production potential of bifidobacterial isolates from ruminal fluid samples of Murrah buffaloes.

In the present study, we investigated the potential of Bifidobacterium spp., isolated from ruminal fluid samples from buffaloes (Bubalus bubalis) for conjugated linoleic acid (CLA) production. A total of 294 isolates were obtained from 86 ruminal fluid samples using Bifidus Selective Medium (BSM) medium, and based on phospoketolase assay, 24 isolates were presumptively confirmed to be Bifidobacterium species. Further, the isolates were confirmed morphologically, biochemically and by PCR assays for genus specific (16s rDNA) and transaldolase genes. All 24 strains were positive for conversion of linoleic acid (LA) to CLA by spectrophotometric screening. Gas chromatographic analysis showed that the strains produced cis9, trans11 and tran10, cis12 CLA isomers in LA-supplemented deMan-Rogosa-Sharpe (MRS) broth. The strains were identified as B. thermophilum (n = 21) and B. pseudolongum (n = 3) based on 16 rDNA sequence analysis. The study shows that Bifidobacterium spp., present in the rumens of buffaloes produce CLA from LA and the strains may have the potential to be used as probiotics to enhance the nutraceutical value of ruminant food products.

Effect of ammonia fiber expansion on the available energy content of wheat straw fed to lactating cattle and buffalo in India.

The seasonal lack of availability of lush green forages can force dairy farmers in developing nations to rely on crop residues such as wheat and rice straw as the major feed source. We tested whether ammonia fiber expansion (AFEX) treatment of wheat straw would increase the energy available to Murrah buffalo and Karan-Fries cattle consuming 70% of their diet as wheat straw in India. Forty lactating animals of each species were blocked by parity and days in milk and randomly assigned to 1 of 4 treatment diets (n = 10). Treatments were a nutrient-rich diet with 0 to 20% straw (positive control; PC) and 3 high-straw diets with various levels of AFEX-treatment: (1) 70% untreated straw (no AFEX), (2) 40 to 45% untreated straw with 25 to 30% AFEX-treated straw (low AFEX), and (3) 20% untreated straw with 50% AFEX-treated straw (high AFEX). The AFEX-treated straw was pelleted. Urea was added to the no and low AFEX diets so they were isonitrogenous with the high AFEX diet. Animals were individually fed the PC diet for 14 d followed by 7 d of adaptation to treatments, full treatments for 28 to 35 d, and finally PC diets for 21 d. Compared with buffalo fed the PC diet, those fed high-straw diets consumed 29% less feed dry matter, put out 16% less milk energy, and lost 0.8 kg/d more body weight; the AFEX treatment of straw did not alter intake or milk production but greatly ameliorated the body weight loss (-1.0 kg/d for no AFEX and -0.07 kg/d for high AFEX). In Karan-Fries cattle, high-straw diets decreased dry matter intake by 39% and milk energy by 24%, and the high AFEX diet increased intake by 42% and milk energy by 18%. The AFEX treatment increased digestibilities of organic matter, dry matter, neutral detergent fiber, acid detergent fiber, and crude protein by 6 to 13 percentage points in buffalo and 5 to 10 points in cattle. In conclusion, AFEX treatment increased the digestibility and energy availability of wheat straw for lactating buffalo and cattle and has commercial potential to improve milk production and feed efficiency when high-quality forages or grains are not available.

Polyunsaturated Fatty Acids in Male Ruminant Reproduction - A Review.

Fatty acids such as n-3 and n-6 polyunsaturated fatty acids (PUFA) are critical nutrients, used to improve male reproductive performance through modification of fatty acid profile and maintenance of sperm membrane integrity, especially under cold shock or cryopreservation condition. Also, PUFA provide the precursors for prostaglandin synthesis and can modulate the expression patterns of many key enzymes involved in both prostaglandin and steroid metabolism. Many studies carried out on diets supplemented with PUFA have demonstrated their capability to sustain sperm motility, viability and fertility during chilling and freezing as well as improving testis development and spermatogenesis in a variety of livestock species. In addition to the type and quantity of dietary fatty acids, ways of addition of PUFA to diet or semen extender is very crucial as it has different effects on semen quality in male ruminants. Limitation of PUFA added to ruminant ration is due to biohydrogenation by rumen microorganisms, which causes conversion of unsaturated fatty acids to saturated fatty acids, leading to loss of PUFA quantity. Thus, many strategies for protecting PUFA from biohydrogenation in rumen have been developed over the years. This paper reviews four aspects of PUFA in light of previous research including rumen metabolism, biological roles, influence on reproduction, and strategies to use in male ruminants.

Dietary supplementation of Butyrivibrio fibrisolvens alters fatty acids of milk and rumen fluid in lactating goats.

BACKGROUND: Conjugated linoleic acid (CLA) isomers have high health amelioration potential and hence it is of great interest to increase the CLA content in dairy products. The present study was conducted to investigate the effect of administration of high CLA producing Butyrivibrio fibrisolvens In-1 on fatty acid composition of milk and rumen fluid in lactating goats. Four groups (n = 5) of lactating goats were assigned the following treatments: Control (C) (basal diet); T1 (basal diet + linoleic acid source), T2 (basal diet + suspension of Butyrivibrio fibrisolvens In-1, 10(9) CFU head(-1)) and T3 (basal diet + linoleic acid source + suspension of Butyrivibrio fibrisolvens In-1, 10(9) CFU head(-1)). RESULTS: Rumen liquor and milk samples were collected on days 0, 15, 30, 60 and 90 of the experiment and linoleic isomerase enzyme (LA-I) activity and fatty acid profiles were elucidated. Major effects of treatments were seen on day 30 of the experiment. Total CLA content of rumen fluid increased (P < 0.05) by 218.72, 182.26 and 304% whereas total saturated fatty acid (SFA) content was lowered (P < 0.05) by 6.1, 4.44 and 9.55% in T1, T2 and T3, respectively, as compared to control. Vaccenic acid in groups T2 and T3 increased (P < 0.05) by 66.67% and 105.7% as compared to control. In milk, total CLA increased by 2.03, 1.61 and 0.61 folds in T3, T2 and T1, respectively. Total monounsaturated fatty acid and polyunsaturated fatty acid content increased (P < 0.05) in group T3 by 14.15 and 37.44%, respectively. CONCLUSION: Results of the present study indicated that administration of B. fibrisolvens In-1 along with a linoleic acid (LA) source is a useful strategy to alter the biohydrogenation pattern in the rumen that subsequently decreased SFA content while increased CLA and unsaturated fatty acids in ruminant's milk.

Characterization of CLA-producing Butyrivibrio spp. reveals strain-specific variations.

Conjugated Linoleic Acid (CLA), a fatty acid with high nutraceutical value is produced in rumen by resident bacterial species, especially Butyrivibrio spp. The present study was undertaken to examine the diversity of indigenous Butyrivibrio spp. from rumen liquor of Indian ruminants. The isolates were screened for their CLA production capability at different level of linoleic acid (LA) (0, 200, 400, 600, 800 µg/ml) at different time intervals (0, 2, 4, 6, 12, and 24 h). A total of more than 300 anaerobic cultures were isolated and 31 of them were identified as Butyrivibrio spp. based on morphological, biochemical and molecular characterization. Further, molecular characterization revealed that a large portion (67.7 %) of isolated Butyrivibrio belonged to Butyrivibrio fibrisolvens (B. fibrisolvens) species which is considered to be the most active bacteria amongst the rumen bacteria populace in terms of CLA production. Bacterial isolate VIII (strain 4a) showed highest CLA production ability (140.77 µg/ml) when incubated at 200 µg/ml LA for 2 h, which is 240 % higher than the isolate XXVII, Butyrivibrio proteoclasticus (B. proteoclasticus) showing lowest CLA production (57.28 µg/ml) amongst the screened isolates. It was evident from the observations recorded during the course of experiments that CLA production ability is strain specific and thus did not follow a single pattern. CLA production also varied with time of incubation and concentration of free linoleic acid supplemented in the growth medium. The results of these findings put forward a strain that is high CLA producer and can be further exploited as an additive for enhancing meat and milk quality in ruminants.

Dietary Inorganic Chromium in Summer-Exposed Buffalo Calves (Bubalus bubalis): Effects on Biomarkers of Heat Stress, Immune Status, and Endocrine Variables.

The aim of this study was to evaluate the effect of different levels of inorganic chromium (Cr) on heat stress, immune response, and hormonal variation in Murrah buffalo calves during the summer season. Twenty-four growing Murrah buffalo calves were randomly allocated into four treatments for a period of 120 days. Feeding regimen was same in all the groups, except the buffalo calves in treatment groups were additionally supplemented with 0.5, 1.0, and 1.5 mg of inorganic Cr/kg dry matter. Buffalo calves were monitored daily for physiological variables and dry matter intake (DMI) and fortnightly for body weight change. Blood samples were collected at day 0, 15, 30, 45, 60, 75, 90, 105, and 120 and analyzed for heat shock protein 70 (Hsp 70), lymphocyte proliferation, neutrophil phagocytic activity, immunoglobulin, ferric reducing antioxidant power (FRAP) assay, insulin, cortisol and thyroid hormones, and Cr levels. Dietary Cr supplementation did not have any effect on DMI, growth performance, and physiological variables. However, lymphocyte proliferation, neutrophil phagocytic activity, plasma immunoglobulin, FRAP value, and plasma Cr concentration increased significantly (P < 0.05) with increase in levels of Cr. Adding Cr to the diet of summer-exposed buffalo calves did not show any effect on plasma levels of thyroid hormone, while concentration of insulin, cortisol, and Hsp 70 decreased (P < 0.05). Supplementation of inorganic Cr to the diet of buffalo calves reared under high ambient temperature improved heat tolerance, immune status without affecting nutrient intake, and growth performance.

Body condition, energy balance and immune status of periparturient Murrah buffaloes (Bubalus bubalis) supplemented with inorganic chromium.

Periparturient Murrah buffaloes were used to determine whether body condition, energy balance and immune status are affected by inorganic Cr supplementation. Twenty-four Murrah buffaloes were blocked into four groups having six animals in each group and fed for 60 days pre-partum to 150 days post-partum. Feeding regimen was same in all the groups except that these were supplemented with 0.0, 0.5, 1.0 and 1.5 of Cr per kilogram of dry matter (DM) in the four respective groups. Buffaloes were weighed at fortnightly intervals. Blood samples were collected from the jugular vein at days -60, -30, -15, -7, 0, 7, 15, 30, 60, 90, 120 and 150 of experimental feeding for the estimation of non-esterified fatty acid (NEFA), beta-hydroxybutyric acid (BHBA), Cr level, lymphocyte proliferation, neutrophil phagocytic activity, plasma total immunoglobulin (TIg), immunoglobulin G (IgG) and cortisol levels. Results revealed that with approaching parturition, dry matter intake (DMI), immune response and plasma Cr level decreased (P < 0.05) gradually and minimum values were observed on the day of parturition in all groups. In contrast, body condition score (BCS), plasma NEFA and BHBA concentrations showed increasing (P < 0.05) trends towards calving and level decreased after calving. Dietary Cr supplementation did not have any effect on DMI and BCS, but immune response and plasma Cr concentration showed a positive correlation with dietary Cr supplementation. Buffaloes supplemented with 1.5 mg/kg Cr had significantly (P < 0.05) low plasma NEFA and BHBA concentrations. The results of present findings indicated that dietary inorganic Cr supplementation reduced lipid mobilization and improved immune response in periparturient buffaloes.

Energy metabolites, lipid variables and lactation performance of periparturient Murrah buffaloes (Bubalus bubalis) fed on diet supplemented with inorganic chromium.

The aim of this study was to elucidate the effects of chromium (Cr) supplementation as inorganic Cr (CrCl3·6H2O) on energy balance, lipid peroxidation, and lactation performance in periparturient Murrah buffaloes. Twenty-four multiparous Murrah buffaloes according to lactation, parity, body mass, and expected calving date were divided equally. Experimental buffaloes were randomly assigned to four treatment diets: a control diet and three diets with an inorganic Cr supplementation at 0.5, 1.0, and 1.5 mg of Cr/kg dry matter (DM), respectively from 60 days before expected calving date until 60 days of lactation. Milk productions of buffaloes were recorded every day until 60 days in milk. Blood samples were collected at days -60, -45, -30,-21, -15, -7, -3, 0, 7, 15, 21, 30, 45, and 60 days relative to actual calving for determination of plasma glucose, nonesterified fatty acid (NEFA), thiobarbituric acid reactive substance (TBARS), total cholesterol, total protein, albumin, blood urea nitrogen (BUN), and minerals. Adding inorganic Cr to the diet of Murrah buffaloes increased milk yield. Percentage of fat and total solid yield increased significantly through the experiment in the Cr-supplemented group. At the day of calving, buffaloes showed a decrease in dry matter intake (DMI), plasma glucose, and zinc (Zn) and Cr concentrations. In contrast, plasma NEFA, TBARS, and copper (Cu) levels were found highest at the day of calving among all groups. Cr supplementation increased peripheral blood glucose concentration while decreased level of NEFA and TBARS was recorded in Cr-fed buffaloes. Supplemental Cr had no effect on plasma cholesterol, total protein, albumin, and BUN in periparturient period. Dietary Cr supplementation had positive effect on plasma Cr concentration, but the plasma concentration of Cu, Zn, and iron (Fe) was not affected by different dietary Cr level supplementation. The results suggest that dietary inorganic Cr supplementation improved milk yield by reducing negative energy balance and lipid peroxidation in buffaloes during periparturient period.

Effect of feeding inorganic chromium on growth performance, endocrine variables, and energy metabolites in winter-exposed buffalo calves (Bubalus bubalis).

We investigated the effect of chromium (Cr) supplementation on the growth performance, energy metabolites, and hormonal variation in winter-exposed buffalo calves. Twenty-four female buffalo calves were randomly allotted to four dietary treatments (n = 6) for a period of 120 days. Feeding regimen was the same in all the groups, except the animals in the four respective groups were additionally supplemented with 0.0, 0.5, 1.0, and 1.5 mg of Cr/kg DM in the form of CrCl3.6H2O. Calves were monitored daily for physiological variables and dry matter intake (DMI). Blood samples were collected at fortnightly intervals from each buffalo calves to measure concentrations of hormones (insulin, cortisol, and growth hormone), energy metabolites (glucose and non-esterified fatty acids), and plasma mineral levels. After 120 days of feeding trial, buffalo calves fed with Cr had lower (P < 0.05) circulating plasma concentrations of glucose, insulin, and cortisol hormones, whereas plasma thyroid hormone and non-esterified fatty acids concentrations were found similar (P > 0.05) among all the treatments. The results suggested that dietary Cr supplementation influenced plasma Cr levels without affecting the plasma concentrations of other trace minerals. However, physiological variables, nutrient intake, and growth performance of buffalo calves did not differ among all treatments (P > 005). In summary, the current study showed that supplementation of Cr at the level of 1.0 and 1.5 mg of Cr/kg DMI was more effective in improving glucose utilization by increasing potency of insulin hormone and reducing concentration of cortisol hormone. Results also suggested that supplemental Cr also improves blood plasma Cr levels.

Assessment of chromium content of feedstuffs, their estimated requirement, and effects of dietary chromium supplementation on nutrient utilization, growth performance, and mineral balance in summer-exposed buffalo calves (Bubalus bubalis).

This study was conducted to determine the chromium content of different feedstuffs, their estimated requirement, and effect of dietary Cr supplementation on nutrient intake, nutrient utilization, growth performance, and mineral balance in buffalo calves during summer season. Levels of Cr was higher in cultivated fodder, moderate in cakes and cereal grains, while straw, grasses, and non-conventional feeds were poor in Cr content. To test the effect of Cr supplementation in buffalo calves, 0, 0.5, 1.0, and 1.5 ppm of inorganic Cr were fed to 24 buffalo calves. Buffalo calves were randomly assigned to four treatments (n = 6) and raised for 120 days. A metabolic trial for a period of 7 days was conducted after 3 months of dietary treatments. Blood samples were collected at fortnight interval for plasma mineral estimation. The results suggested that dietary Cr supplementation in summer did not have any affects (P > 0.05) on feed consumption, growth performance, nitrogen balance, and physiological variables. However, dietary Cr supplementation had significant effect (P < 0.05) on balance and plasma Cr (ppb) levels without affecting (P > 0.05) balance and plasma levels of other trace minerals. The estimated Cr requirement of buffalo calves during summer season was calculated to be 0.044 mg/kg body mass and 10.37 ppm per day. In conclusion, dietary Cr supplementation has regardless effect on feed consumption, mass gain, and nutrient utilization in buffalo calves reared under heat stress conditions. However, supplementation of Cr had positive effect on its balance and plasma concentration without interacting with other trace minerals.