Autochthonous buffalo-gut origin Pediococcus pentosaceus RM119 decreased diarrhea and enhanced gut health, immunity in neonatal Murrah calves.

This study assessed the effect of autochthonous probiotic Pediococcus pentosaceus RM119 on gut health, growth, and nutrient utilization in calves. Twelve buffalo calves (<15 d) were divided into two groups, control without probiotics, and probiotic group with P. pentosaceus RM119 @ 108 CFU/calf/d. The probiotic group showed a reduction (p < 0.05) in fecal score, diarrhea episodes and duration of diarrhea. The fecal pH, fecal ammonia was lower, whereas lactate was higher in probiotic group than control. There was a significant increase (p < 0.01) in the concentration of fecal acetate, propionate and butyrate levels in the probiotic supplemented group. The fecal lactobacilli and bifidobacterium were higher (p < 0.01), whereas, fecal coliform and clostridial count were lower (p < 0.01) in P. pentosaceus RM119 supplemented group. There was an improvement in reduced glutathione anti oxidant. Overall, buffalo-gut origin P. pentosaceus RM119 reduced the frequency and severity of diarrhea in neonatal buffalo calves and improved the gut health.

Microencapsulated and Lyophilized Lactobacillus acidophilus Improved Gut Health and Immune Status of Preruminant Calves.

The present study was conducted to study the effect of microencapsulated, lyophilized, or fermented milk using Lactobacillus acidophilus NCDC15 as a probiotic to improve gut health, growth, nutrient utilization, and immunity status of young crossbred calves. The viable culture of L. acidophilus was used for preparation of different probiotic forms/products. To compare the efficacy of probiotic products, twenty crossbred calves (3-day old) were divided into four groups (n = 5), control (C), fed only milk and basal diet, and treatment groups, supplemented with microencapsulated, fermented, and lyophilized probiotic at 108 colony-forming units, respectively. Probiotic-supplemented groups showed reduction in faecal score, faecal pH, and ammonia concentration as compared to control indicating decreased diarrheal incidence. There was an increase (P < 0.05) in the concentration of faecal lactate and butyrate in the probiotic-supplemented groups. The faecal count (log10 (CFU)/g of fresh faeces) of lactobacilli and bifidobacteria was higher (P < 0.05), whereas faecal coliforms and clostridia count were reduced (P < 0.001) in all the probiotic fed groups as compared to control. The cell-mediated immunity was improved (P < 0.05) in the microencapsulated and fermented probiotic groups. However, there was no effect on the nutrient utilization, average daily gain, and blood biochemical profile. Therefore, it is concluded that the fermented, microencapsulated and lyophilized probiotic products were superior in improving the gut health in terms of its microbiota and metabolites and cell-mediated immunity response in calves, irrespective of form of probiotic. The increased population of Lactobacillus and Bifidobacterium decreased the colonization of the gut by pathogens such as Escherichia coli and Clostridium by exclusion and production of organic acids in the intestine. This decreased the diarrhoeal incidence (1.3 vs 1.8) and days in diarrhoea (3.9 vs 5.8) in calves in probiotic fed groups as compared to control.

Insights into Metatranscriptome, and CAZymes of Buffalo Rumen Supplemented with Blend of Essential Oils.

The aim of this study was to investigate the rumen microbial diversity and functionality in buffaloes fed with a blend of essential oils (BEO) using LSD switch over design. The BEO consisting of blend of Trachyspermum copticum (Ajwain) oil, Cymbopogon citratus (lemon grass) oil and Syzygium aromaticum (clove bud) oleoresin mixed in equal proportion, was fed at the rate of 0, 0.75 and 1.5 ml/100 kg of body weight in 0 (control), 0.75 and 1.5 groups, respectively. The metatranscriptomic libraries of the rumen microbiome were represented by 7 domains, 84 phyla, 64 archeal genera and 663 bacterial genera with Bacteroidetes and Firmicutes constituting 80% of phyla abundance irrespective of feeding regime. Methanogenic archaea was represented by 22 phyla with Methanobrevibacter as the major genus. BEO feeding reduced the abundance of Methanococcus and Thermoplasma (P < 0.05) at all levels. The results revealed that the feeding of BEO shifted the archeal and bacterial population at very low magnitude. The study explored the vast diversity of buffalo rumen bacteria and archaea, and the diverse wealth of rumen enzymes (CAZymes), which revealed that a major part of CAZymes comes from the less known rumen microbes indicating alternative paths of fiber degradation along with the very well known ones.

Effect of feeding of blend of essential oils on methane production, growth, and nutrient utilization in growing buffaloes.

OBJECTIVE: An experiment was conducted to study the effect of a blend of essential oils (BEO) on enteric methane emission and growth performance of buffaloes (Bubalus bubalis). METHODS: Twenty one growing male buffaloes (average body weight of 279±9.3 kg) were divided in to three groups. The animals of all the three groups were fed on a ration consisting of wheat straw and concentrate mixture targeting 500 g daily live weight gain. The three dietary groups were; Group 1, control without additive; Group 2 and 3, supplemented with BEO at 0.15 and 0.30 mL/kg of dry matter intake (DMI), respectively. RESULTS: During six months feeding trial, the intake and digestibility of dry matter and nutrients (organic matter, crude protein, ether extract, neutral detergent fibre, and acid detergent fibre) were similar in all the groups. The average body weight gain was tended to improve (p = 0.084) in Group 2 and Group 3 as compared to control animals. Feeding of BEO did not affect feed conversion efficiency of the animals. The calves of all the three groups were in positive nitrogen balance with no difference in nitrogen metabolism. During respiration chamber studies the methane production (L/kg DMI and L/kg digestible dry matter intake was significantly (p<0.001) lower in Group 2 and Group 3 as compared to control animals. CONCLUSION: The results indicated that the BEO tested in the present study have shown potential to reduce enteric methane production without compromising the nutrient utilization and animal performance and could be further explored for its use as feed additive to mitigate enteric methane production in livestock.

Effect of high and low roughage total mixed ration diets on rumen metabolites and enzymatic profiles in crossbred cattle and buffaloes.

AIM: A comparative study was conducted on crossbred cattle and buffaloes to investigate the effect of feeding high and low roughage total mixed ration (TMR) diets on rumen metabolites and enzymatic profiles. MATERIALS AND METHODS: Three rumen-fistulated crossbred cattle and buffalo were randomly assigned as per 3×3 switch over design for 21-days. Three TMR diets consisting of concentrate mixture, wheat straw and green maize fodder in the ratios of (T1) 60:20:20, (T2) 40:30:30, and (T3) 20:40:40, respectively, were fed to the animals ad libitum. Rumen liquor samples were collected at 0, 2, 4, 6, and 8 h post feeding for the estimation of rumen biochemical parameters on 2 consecutive days in each trial. RESULTS: The lactic acid concentration and pH value were comparable in both species and treatments. Feed intake (99.77±2.51 g/kg body weight), ruminal ammonia nitrogen, and total nitrogen were significantly (p<0.05) higher in buffalo and in treatment group fed with high concentrate diet. Production of total volatile fatty acids (VFAs) was non-significant (p>0.05) among treatments and significantly (p<0.05) greater in crossbred cattle than buffaloes. Molar proportions of individual VFAs propionate (C3), propionate:butyrate (C3:C4), and (acetate+butyrate):propionate ([C2+C4]:C3) ratio in both crossbred cattle and buffalo were not affected by high or low roughage diet, but percentage of acetate and butyrate varied significantly (p<0.05) among treatment groups. Activities of microbial enzymes were comparable among species and different treatment groups. A total number of rumen protozoa were significantly (p<0.05) higher in crossbred cattle than buffaloes along with significantly (p<0.05) higher population in animal fed with high concentrate diet (T1). CONCLUSION: Rumen microbial population and fermentation depend on constituents of the treatment diet. However, microbial enzyme activity remains similar among species and different treatments. High concentrate diet increases number of rumen protozoa, and the number is higher in crossbred cattle than buffaloes.

Impact of levels of total digestible nutrients on microbiome, enzyme profile and degradation of feeds in buffalo rumen.

The present study was aimed at understanding a shift in rumen microbiome of buffaloes fed various levels of total digestible nutrients. To understand the process, the metagenomics of rumen microbes, in vivo and in vitro rumen fermentation studies were carried out. Three rumen fistulated adult male Murrah buffaloes were fed three isonitrogenous diets varying in total digestible nutrients (70, 85 and 100% of TDN requirement) in 3X3 switch over design. On dry matter basis, wheat straw/ roughage content were 81, 63 and 51% and that of maize grain was 8, 16 and 21% in three diets respectively. After 20 d of feeding, rumen liquor and rumen contents were sampled just before (0h) and 4h post feeding. Ruminococcus flavefaciens and R. albus (estimated with real time PCR) were higher in high roughage diets. The predominant phyla in all the three groups were Bacteroidetes, Firmicutes followed by Proteobacteria, Actinobacteria and Fibrobacteres. A core group of more than fifty rumen bacteria was present in all the animals with very little variations due to level of TDN. The most predominant bacterial genera reported in order of decreasing abundance were: Prevotella, Bacteroides, Clostridium, Ruminococcus, Eubacterium, Parabacteroides, Fibrobacter, Butyrivibrio etc. The higher diversity of the enyzmes families GH 23, GH 28, GH 39, GH 97, GH 106, and GH 127 (the enzymes active in fibre and starch degradation) were significantly higher on 100%TDN diet while CE 14 (required for the hydrolysis of bond between carbohydrate and lignin) was higher on low TDN (70%) diet, indicating ester bond cleavage was better in animals fed high roughage (wheat straw) diet.

Effect of feeding tannin-degrading bacteria Streptococcus gallolyticus strain TDGB 406 on meat quality of goats fed with Quercus semicarpifolia leaves.

The effect of feeding tannin-degrading bacteria (Streptococcus gallolyticus strain TDGB 406) on carcass characteristics of goats fed with oak (Quercus semicarpifolia) leaves was studied on 18 male goats (4 months old, average body weight 9.50 ± 1.50 kg), distributed into three groups of six animals each. The animals of group 1 served as control, while the animals of groups 2 and 3 were given (at 5 ml/kg live weight) autoclaved and live culture of isolate TDGB 406 (10(6) cells/ml), respectively. The animals were fed with oak leaves as a basal roughage source and maize hay along with fixed quantity of concentrate mixture. After 4 months of feeding, the animals were slaughtered for carcass studies. The feeding of live culture of isolate TDGB 406 did not cause any effect (P > 0.05) on pre-slaughter weight, empty body weight, carcass weight, dressing percent, and yield of wholesale cuts (neck, rack, shoulder, breast, shank, loin, leg, and flank) of the goat meat. The chemical composition of longissimus dorsi muscle was comparable (P > 0.05) among the groups. The organoleptic evaluation of pressure-cooked meat in terms of tenderness and overall palatability was increased significantly (P < 0.05) in the meat of group 3 where live culture was supplemented. The other attributes were similar among the groups. It was concluded that supplementation of tannin-degrading bacteria S. gallolyticus strain TDGB 406 to goats fed with oak leaves did not affect the carcass characteristics and meat quality.

Effect of feeding tannin degrading bacterial culture (Streptococcus gallolyticus strain TDGB 406) on nutrient utilization, urinary purine derivatives and growth performance of goats fed on Quercus semicarpifolia leaves.

To study the effect of supplementation of tannin degrading bacterial culture (Streptococcus gallolyticus strain TDGB 406) on growth performance, nutrient utilization and urinary purine derivatives of goats fed on oak (Quercus semicarpifolia) leaves. For growth study, eighteen billy goats (4 month old, average body weight 9.50 ± 1.50 kg) were distributed into three groups of six animals each. The animals of group 1 served as control while animals of groups 2 (T1) and 3 (T2) were given (@ 5 ml/kg live weight) autoclaved and live culture of isolate TDGB 406 (10(6) cells/ml) respectively. The animals were fed measured quantity of dry oak leaves as the main roughage source and ad libitum maize hay along with fixed quantity of concentrate mixture. The feeding of live culture of isolate TDGB 406 (probiotic) did not affect dry matter intake and digestibility of nutrients except that of dry matter and crude protein, which was higher in T2 group as compared to control. All the animals were in positive nitrogen balance. There was no significant effect of feeding isolate TDGB 406 on urinary purine derivatives (microbial protein production) in goats. The body weight gain and average live weight gain was significantly higher (p = 0.071) in T2 group as compared to control. Feed conversion efficiency was also better in the goats fed on live culture of TDGB 406 (T2). The feeding of tannin degrading bacterial isolate TDGB 406 as probiotic resulted in improved growth performance and feed conversion ratio in goats fed on oak leaves as one of the main roughage source.

Eubacterium pyruvativorans sp. nov., a novel non-saccharolytic anaerobe from the rumen that ferments pyruvate and amino acids, forms caproate and utilizes acetate and propionate.

Two similar gram-positive rods were isolated from 10(-6) dilutions of ruminal fluid from a sheep receiving a mixed grass hay/concentrate diet, using a medium containing pancreatic casein hydrolysate as sole source of carbon and energy. The isolates did not ferment sugars, but grew on pyruvate or trypticase, forming caproate as the main fermentation product and valerate to a lesser extent. Acetate and propionate were utilized. One of these strains, I-6T, was selected for further study. Strain I-6T was a non-motile coccal rod, 1.2 x 0.4 microm, with a gram-positive cell wall ultrastructure and a G + C content of 56.8 mol%. No spores were visible, and strain I-6T did not survive heating at 80 degrees C for 10 min. Its rate of NH3 production was 375 nmol (mg protein)(-1) min(-1), placing it in the 'ammonia-hyperproducing' (or HAP) group of ruminal bacteria. 16S rDNA sequence analysis (1296 bases) indicated that it represents a novel species within the 'low-G + C' gram-positive group, for which the name Eubacterium pyruvativorans sp. nov. is proposed. Among cultivated bacteria, strain I-6T was most closely related (89% identity) to other asaccharolytic Eubacterium isolates from the mouth and the rumen. It was 98% identical to uncultured bacterial sequences amplified by others from ruminal digesta.

Microbial status and rumen enzyme profile of crossbred calves fed on different microbial feed additives.

AIMS: To test various microbial cultures as cattle feed additives. METHODS AND RESULTS: Four groups of newly born crossbred calves (average body weight 23.5 kg) were reared on green berseem and calf starter which was devoid of cereal grains. Milk was fed up to 8 weeks of age, starting with one tenths and gradually reducing to one twentieths of the body weight. One hundred millilitres of microbial feed additive or 100 g fermented feed was fed to the animals of group 2 (curd containing lactic acid bacteria at 10(8) cfu x ml(-1)), group 3 (Saccharomyces cerevisiae NCDC-49 at 10(6) cfu x ml(-1)) and group 4 (Lactobacillus acidophilus-15 at 10(8) cfu x ml(-1)). Group 1 served as control. The incidence and duration of diarrhoea was lower in the animals of probiotic fed groups as compared to control group. Out of three microbial feed additives, yeast feeding showed maximum suppression of diarrhoea followed by Lactobacillus and curd. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: There was no effect of probiotic feeding on the log number of cells of lactic acid bacteria, yeast and coliform bacteria in the faeces and rumen liquor at any age. The activities of carboxymethylcellulase, xylanase, beta-glucosidase, alpha-glucosidase, alpha-amylase, protease, urease and pH of the rumen liquor remained unaffected by probiotic feeding at all ages tested in this experiment.

Selection of Saccharomyces cerevisiae strains for use as a microbial feed additive.

Saccharomyces cerevisiae ITCCF 2094, NCIM 3052, 1031, 1032, NCDC 42, 45, 47, 49 and 50 were screened for their tolerance to pH 2.0-7.0, various concentrations (0.00, 0.10, 0.25 0.50 and 1.0%) of a mixture of acetic, propionic and butyric acids (70:20:10), and bile salts (0.00, 0.30, 0.60 and 0.90%). Low pH (2.0-4.0) and addition of organic acids or bile salts in the medium inhibited the growth of all the strains tested, but the percentage of inhibition was variable in the different strains of yeast. Two of the strains showing maximum tolerance, 42 and 49, were further tested for in vitro dry matter degradability (IVDMD) using green berseem, wheat straw and oat hay as substrates. Saccharomyces cerevisiae 49 enhanced the IVDMD of berseem and wheat straw whereas S. cerevisiae 42 was ineffective. Based on the results of the present experiment, S. cerevisiae NCDC 49 can be considered as the best strain which might tolerate the adverse conditions in the gastrointestinal tract when used as a live microbial feed supplement in the diet of the animals.

Influence of 1-[(E)-2-(2-methyl-4-nitrophenyl)diaz-1-enyl]pyrrolidine-2-carboxylic acid and diphenyliodonium chloride on ruminal protein metabolism and ruminal microorganisms.

The effects of 1-[(E)-2-(2-methyl-4-nitrophenyl)diaz-1-enyl]pyrrolidine-2-carboxy lic acid (LY29) and diphenyliodonium chloride (DIC) on the degradation of protein to ammonia were determined in a mixed rumen microbial population taken from sheep on a grass hay-concentrate diet. Both compounds decreased NH3 production by inhibiting deamination of amino acids. LY29, but not DIC, inhibited growth of the high-activity ammonia-producing species, Clostridium aminophilum and Clostridium sticklandii.