Non-encapsulated, encapsulated, and lyophilized probiotic Limosilactobacillus reuteri SW23 influenced the growth and gut health in calves.

The present study was conducted to assess the impact of non-encapsulated, air-dried microencapsulated, and lyophilized microencapsulated probiotics in indigenous cattle calves (Bos indicus). Twenty-four (5-7 days old) indigenous cattle calves were selected and assigned into four groups, with six calves in each as follows: control (CON), fed milk and basal diet alone, and treatment groups supplemented with non-encapsulated (NEC), air-dried microencapsulated (AEC) and lyophilized microencapsulated (LEC) probiotic L. reuteri SW23 at 108 CFU/head/day in skim milk as a carrier provided for 60 days. The animals were divided into four groups, adopting a complete randomized design, and the effects were considered significant at p ≤ 0.05. Probiotics supplementation increased (p < 0.05) body weight gain (kg), average daily gain, and structural growth measurements in calves of all treatment groups. Dry matter intake (g/d), feed conversion efficiency, and fecal counts of Lactobacilli and Bifidobacteria were also increased in the treatment groups compared to CON. The fecal consistency index was highest in CON (0.70 ± 0.03), followed by NEC (0.68 ± 0.01), AEC (0.66 ± 0.02), and LEC (0.65 ± 0.02). Fecal pH and ammonia levels were reduced (p < 0.05) in the probiotic-fed groups compared to CON, with a concomitant increase in fecal lactate, acetate, and propionate levels. In addition, cell-mediated and humoral immunity were significantly increased in supplemented groups as compared to CON. Thus, it can be concluded that supplementation of the probiotics in microencapsulated/non-encapsulated forms to neonatal calves had a variety of positive effects on their health, including better performance, improved gut health, and a lower fecal consistency index. Moreover, among all supplemented groups, the lyophilized microencapsulated group outperformed air-dried microencapsulated and non-microencapsulated groups in terms of ADG, DMI, and gut health.

Comparative study of depotash vinasse and molasses as binder and production performance in lactating Murrah buffaloes.

Dairy sector has recently focused a lot of attention on the addition of agricultural by-products as functional feed additives as an environmentally friendly and sustainable technology. Depotash vinasse (DPV) serves as a cheap source of nutrients and a binder for animal feed in dairy sector. However, there is little information available on the usage of depotash vinasse on animals. Therefore, the aim of the present study was to assess the role of depotash vinasse as pellet binder on nutrient digestibility, blood parameters and milk production in early lactating Murrah buffaloes. Fifteen Murrah buffaloes (daily milk yield 8.5 to 9.0 kg/day) were randomly assigned to three groups, viz., control, group 1 (G1) and group 2 (G2) on the basis of milk yield and days in milk. The control group animals received a basal diet of concentrate mix, oat greens and wheat straw, G1 animals received molasses as a binder (8%), while G2 received DPV as binder (8%). Results revealed that there was no significant effect on nutrient digestibility. Blood parameters and hepatic enzymes were statistically similar (P > 0.05). Supplementation of depotash vinasse as binder had no effect on plasma minerals and was comparable to control group. There were no changes in milk production and 6% fat-corrected milk yield in treated groups as compared to control. It was concluded that depotash vinasse (8%) may be used for pellet production with no negative impact on milk yield and composition, nutrient digestibility and blood biochemical parameters in early lactating buffaloes.

Techno-functional characterization of fecal lactobacilli isolates of Bos indicus calves for probiotic properties.

In this study, 105 bacterial colonies were isolated from the feces of newborn healthy Bos indicus calves and 37 isolates were confirmed using morphological, biochemical tests, and genus-specific PCR as lactobacilli. 11 isolates were then short-listed for in vitro probiotic testing based on their ability to dwell under acid and bile stress. Species-level identification using 16S rRNA gene sequencing revealed that they were Ligilactobacillus salivarius. These isolates flourished in 0.4% phenol, depicting resistance in adverse conditions encountered in the gastrointestinal tract. The results of cell surface hydrophobicity were found to be 74.50% for RBL12 and 62.62% for RBL09 in hexadecane and xylene, respectively, and that of auto-aggregation was highest in RBL26 (58.92%). These isolates also produced digestive enzymes like amylase, protease, and ß-galactosidase. Further assays reiterated their antimicrobial and coaggregation potential against diarrhea-causing pathogens like Escherichia coli ATCC-25922 and Salmonella arizonae ATCC-13314. Biosafety assessment revealed that none of the tested isolates were hemolytic and mucinolytic in nature. Furthermore, the antioxidant potential of the isolates was also confirmed using 1,1­diphenyl­2­picrylhydrazyl (DPPH) and ferric ion-reducing antioxidant power (FRAP) assay. Along with efficient utilization of inulin, isolates showed promising adhesion ability to the HT-29 cell line. The current findings hence conclude that these Lactobacillus isolates can be exploited as animal probiotics for potential application in young calves to foster gut health and immunity.

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.

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.

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.

Synbiotic formulation of Cichorium intybus root powder with Lactobacillus acidophilus NCDC15 and Lactobacillus reuteri BFE7 improves growth performance in Murrah buffalo calves via altering selective gut health indices.

The objective of this study was to evaluate the effects of synbiotic formulation of Cichorium intybus root powder (C) with Lactobacillus acidophilus NCDC15 (LA) and Lactobacillus reuteri BFE7 (LR) on growth performance in Murrah buffalo calves via monitoring selective gut health indices. Twenty-four Murrah buffalo calves of 5-7 days old and 33 ± 2.0 kg of body weight were distributed randomly into three groups adopting complete randomized design (CRD) as follows: (1) group I served as control (CON) provided with a basal diet alone; (2) group II supplemented with synbiotic formulation of 200 mL L. acidophilus NCDC15 fermented milk with 8 g of Cichorium intybus root powder (LAC) along with basal diet; (3) group III supplemented with synbiotic formulation of 200 mL L. reuteri BFE7 fermented milk with 8 g of Cichorium intybus root powder (LRC) along with basal diet. The final body weight (BW), average dry matter intake (DMI) and structural body measurements were significantly increased (P < 0.05) in LAC and LRC groups by synbiotic as compared to the CON. No effect was registered on apparent nutrient digestibility coefficient of various nutrients in supplemented groups. Faecal score was reduced by the supplementation of synbiotic being lowest in LRC followed by LAC and CON. Calves supplemented synbiotic showed lower (P < 0.05) faecal pH and ammonia with a concomitant increase in faecal lactate levels and faecal short chain fatty acids (SCFA) as compared to control. The faecal Lactobacillus and Bifidobacterium population was increased (P < 0.05) in synbiotic fed groups as compared to control. Additionally, coliform and clostridia count was decreased (P < 0.05) in treatment groups compared to CON. Overall, it may be concluded that synbiotic supplementation was effective in improving the growth performance in Murrah buffalo calves via altering selective gut health indices.

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.

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.

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.

Assessment of feeding varying levels of Metabolizable energy and protein on performance of transition Murrah buffaloes.

Fifteen close up pregnant Murrah buffaloes of mean body weight (668.3 ± 24.03) kg, lactation number (2.8 ± 0.17) and expected producing ability (EPA) (2125.7 ± 46.34) were randomly distributed into three groups each of five animals to investigate the performance at different levels of metabolizable energy and protein. Control group was fed as per ICAR Nutrient requirements of animals (2013) recommendation whereas treatment group (1) high metabolizable energy and high metabolizable protein (HMEMP) and group (2) low metabolizable energy and low metabolizable protein (LMEMP) were offered with ration containing 15% more and 15% less ME and MP, respectively. The feeding trial was carried out for the period of 40 days before parturition and continued for 120 days after parturition. Intake of dry matter (DM) (%BW) was similar among experimental groups. Metabolizable energy (ME) (MJ/100 kg BW) and metabolizable protein (MP) (g/100 kg BW) intake was highest in HMEMP followed by control and LMEMP group, respectively. Digestibility trial of 7 days was conducted at 60 days post-partum and it was observed that apparent digestibility coefficients (%) of DM, organic matter (OM), crude protein (CP), ether extracts (EE), neutral detergent fiber (NDF) and acid detergent fiber (ADF) were similar among the experimental groups. Milk yield (kg/kg DMI) was similar among treatment groups whereas 6% fat corrected milk (FCM) was lower in LMEMP group as compared to HMEMP and control. No significant effect of dietary MP and ME levels on milk composition was observed among experimental groups. There were no significant difference in non esterified fatty acid (NEFA), blood urea nitrogen(BUN), growth hormone (GH) and insulin like growth factor-1(IGF-1) concentration among different experimental groups whereas concentration of immunoglobulin G (IgG) (µg/ml) was found to be lower in LMEMP. The study results indicate that nutrient digestibility and lactation performance was not affected with 15% variation in intakes of ME and MP in lactating Murrah buffaloes.

Effect of omega-3 and omega-6 polyunsaturated fatty acid enriched diet on plasma IGF-1 and testosterone concentration, puberty and semen quality in male buffalo.

The objective of the present study was to evaluate the effect of omega-3 and omega-6 PUFA enriched diet on plasma IGF-1 and testosterone concentrations, puberty, sperm fatty acid profile and semen quality in male buffalo. Eighteen male buffalo calves were distributed randomly in three different groups and fed concentrate mixture along with green fodder and wheat straw in 50:40:10 ratios as per requirements. Basis ration of animals in group I was supplemented with 4% of prilled fat (PFA), while in group II and group III were added 4.67% of Calcium salt from Soybean (CaSFA) and Linseed oil (CaLFA), respectively. Male buffalo fed omega-3 PUFA high diet significantly increased concentrations of IGF-1 and testosterone in plasma as compared to two other diets (p<0.05). The age of puberty and scrotal circumference significantly increased by dietary fat effect (p<0.05) of which n-3 PUFA enriched diet (CaLFA) had the largest influence as compared to other diets (PFA and CaSFA). Feeding of n-3 PUFA rich diet significantly increased the DHA (C22:6n-3) content in sperm (p<0.05), which contributed to increased fluidity of plasma membrane, elevated quality of sperm (motility, viability) and in vitro fertility (plasma membrane integrity, acrosome integrity) in both fresh and post-thawing semen. These findings indicate that feeding of n-3 PUFA enriched diet increased IGF-1 and testosterone secretion, reduced pubertal age and improved both fresh and post-thawing semen quality in male buffalo.

Effect of bypass fat supplementation on productive and reproductive performance in crossbred cows.

Objectives of the present study were to evaluate the effects of supplementation of bypass fat on milk production and reproductive performance of crossbred cows. Nineteen multiparous crossbred cows (2-4 lactation) were divided in two groups on the basis of most probable production ability (MPPA). The animals in group 1 (nine cows, MPPA 3,441.32 kg, control group G1) were fed chaffed wheat straw, chopped green maize, and concentrate mixture as per requirements while the animals in group 2 (10 cows, MPPA 3,457.2 kg, treatment group G2) were fed the same ration supplemented with 2.5% bypass fat (on DMI basis). The cows of G2 were supplemented bypass fat 40 days prepartum to 90 days postpartum and carry over effect of supplementation on milk production and reproductive parameters was monitored up to 210 days of lactation. Average birth weights of the calves were 24.94 and 27.95 kg in G1 and G2, respectively. The calving per cent in G1 (88.88%) was lower than that of G2 (100%). The time taken for expulsion of fetal membranes was decreased (P<0.05) by 5.4 h in G2 compared to G1. Days required for involution of uterus was less (P<0.05) in G2 (35.40 days) than that of G1 (49.44 days). Less number of cases of retention of fetal membranes (RFM) and metritis were observed in G2 as compared to that of G1. The average milk yield (210 days) in G2 was higher (P<0.05) than that of G1 (18.65 vs 17.57 kg/day). Similarly, the milk yield during the carry over period (90 days) was higher (P<0.05) in G2 than that of G1 (14.81 vs 13.79 kg/day). The time required for onset of cyclicity was reduced (P<0.05) by 6.5 days in G2 as compared to that of G1. The service period and AI per conception were also reduced (P<0.05) in G2 while bypass fat feeding showed no effect on conception rate (P<0.80). There were no differences between the two groups in plasma cholesterol, progesterone and insulin concentrations. Hence, it was concluded that bypass fat supplementation at 2.5% of DMI increased the milk production and reduced the time required for involution of uterus and commencement of cyclicity. AI per conception, incidences of metritis and RFM were also reduced on supplementing bypass fat.