Comprehensive transcriptional profiling and functional study of lymphangiogenic growth factor in placentome of early-stage pregnant water buffalo.

The aim of the present study was to evaluate the expression and localization of lymphangiogenic factors (VEGF-C and VEGF-D), their receptor (VEGFR3) and lymphatic endothelial marker (LYVE1) in buffalo placenta during early pregnancy [EP], and to investigate the functional role of lymphangiogenic growth factors in placental lymphangiogenesis. The mRNA and protein expression of VEGF-C, VEGF-D, their receptor VEGFR3 and LYVE1 showed significant expression in EP1 (29-42 days) and EP2 stages (51-82 days) both in caruncle (maternal part) and cotyledon (foetal part) of the buffalo placenta. Immunoreactivity of VEGF-C, VEGF-D and LYVE1 was observed around the endometrial gland, in lymphatics and trophoblast cells, whereas VEGFR3 mainly localized in lymphatics of the caruncle and cotyledons. Cultured trophoblast cells were treated with VEGF-C/VEGF-D (50, 100 and 150 ng/ml) and combined doses of VEGF-C and VEGF-D (150 ng/ml) each for different time durations (24, 48 and 72 h). The mRNA expression of LYVE1 and PCNA was significantly (p < .001) upregulated with VEGF-C and VEGF-D and combined treatment (@150 ng/ml), as well as significantly downregulating Caspase-3 at 48 and 72 h. Thus, the present study provides evidence that lymphangiogenic factors are expressed in buffalo placental compartments and they may play a significant role in the regulation of placental function in water buffaloes.

Host genetics associated with gut microbiota and methane emission in cattle.

In livestock sector, dairy animals alone produce 18% of the total greenhouse gas emissions globally as methane (CH4). This Enteric methane is the largest component of total carbon footprints produced by livestock production system and its reduction is today's new challenge to make livestock farming sustainable for earth's environment. The production of enteric methane in ruminants is a complex phenomena involving different host factors like host genotype, rumen microbiome, host physiology along with dietary factors. Efforts have been made to reduce methane emissions largely through nutritional interventions and dietary supplements, but permanent reductions can be obtained through genetic means by selecting and breeding of low methane emitting animals. From genome-wide association studies, many important genomic QTL regions and single nucleotide polymorphisms involved in shaping the composition of the ruminal microbiome and thus their carbon footprints have been recognised, implying that methane emission traits are quantitative traits. The major bottleneck in implementation of reduced methane emission traits in the breeding programs is wide variation at phenotypic level, lack of precise methane measurements at individual level. Overall, the heritability for CH4 production traits is moderate, and it can be used in breeding programmes to target changes in microbial composition to reduce CH4 emission in the dairy industry for far-reaching environmental benefits at the cost of a minor reduction in genetic gain in production traits.

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.

Effect of supplementing sulphate-reducing bacteria along with sulphur on growth performance, nutrient utilization and methane emission in goats.

A competitive relationship exists between sulphate-reducing bacteria and methanogens in the anaerobic environment including rumen for hydrogen where sulphate is not limiting growth and consequently inhibit enteric methane emission as thermodynamically energetic sulphate reduction (∆Go = - 21.1 kJ/mole of H2) is more favourable than methanogenesis (∆Go = - 16.9 kJ/mole H2). To validate this hypothesis, a study was designed to investigate the effect of supplementation of sulphate-reducing bacteria (SRB) identified as Streptococcus caviae RM296 as microbial feed additives alone or along with sulphur (as sodium sulphate) on methane production, live weight gain, feed intake, nutrient digestibility and energy metabolism in goats. The experiment was conducted on growing kids (n = 36, 5-6 months of age) with average body weight of 10.08 ± 0.21 kg, divided into six groups (n = 6). The duration of the feeding trial was of 150 days. The six treatments were control fed a basal diet (T1), SRB 0.5 ml/kg BW (T2), sulphur (as sodium sulphate) 0.095% of DMI (total sulphur level in the diet 1.5 times the requirement) (T3), sulphur (as sodium sulphate) 0.095% of DMI + SRB 0.5 ml/kg BW (T4), sulphur (as sodium sulphate) 0.19% of DMI (total sulphur level in the diet 2 times the requirement) (T5) and sulphur (as sodium sulphate) 0.19% of DMI + SRB 0.5 ml/kg BW (T6). Duration of study was 150 days and goats were fed as per ICAR (2013) feeding standard. Methane (CH4) production (l/kg DMI) was reduced by 11.8% (P = 0.052) in T6 where sulphur (0.19% DMI) was supplemented along with SRB4 (at the rate 0.5 ml/kg BW) as compared to T1 (un-supplemented group). However, the dry matter intake (DM), total weight gain (TG), average daily gain (ADG), feed conversion ratio (FCR), excretion of purine derivatives (allantoin, uric acid, xanthine and hypoxanthine) and digestibility of organic matter (OM), dry matter (DM), ether extract (EE), crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) were similar (P > 0.05) among all the groups. The experimental data revealed that feeding of SRB as a microbial feed additive along with sulphur (as sodium sulphate) is capable of reducing enteric CH4 emission without any adverse effect on rumen fermentation and digestibility of the nutrients.

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.

Early-Life Intervention of Lactoferrin and Probiotic in Suckling Piglets: Effects on Immunoglobulins, Intestinal Integrity, and Neonatal Mortality.

The aim of this study was to determine the effects of early-life bovine lactoferrin and host specific probiotic interventions on growth performance, mortality, and concentrations of immunoglobulin A and immunoglobulin G and transforming growth factor beta 1 (a marker of intestinal integrity) in serum of neonatal piglets. A total of eight piglet litters from parity matched sows were randomly divided into four groups and assigned to one of the four interventions: control (sterile normal saline), bovine lactoferrin (100 mg bovine lactoferrin), probiotic (1 × 109 colony forming unit (cfu) of swine origin Pediococcus acidilactici FT28 probiotic), and bovine lactoferrin + probiotic (100 mg bovine lactoferrin and 1 × 109 CFU of P. acidilactici FT28 probiotic). All the interventions were given once daily through oral route for first 7 days of life. The average daily gain (p = 0.0004) and weaning weight (p < 0.0001) were significantly improved in the probiotic group. The piglet survivability was significantly higher in bovine lactoferrin and probiotic groups than control group in Log-rank (Mantel-Cox) test. The concentrations of immunoglobulin A on day 21 in bovine lactoferrin, probiotic, and bovine lactoferrin + probiotic groups increased significantly (p < 0.05). Immunoglobulin G concentrations on day 7 and 15 in bovine lactoferrin and bovine lactoferrin + probiotic groups and on day 15 in probiotic group were significantly (p < 0.05) elevated, whereas, the concentration of transforming growth factor-ß1 was significantly (p < 0.05) increased from day 7 to 21 in all the supplemented groups. In conclusion, the early-life bovine lactoferrin and P. acidilactici FT28 probiotic interventions reduced the mortality in the suckling piglets by promoting the systemic immunity and enhancing the intestinal integrity.

Efficacy of Microencapsulated Probiotic as Adjunct Therapy on Resolution of Diarrhea, Copper-Zinc Homeostasis, Immunoglobulins, and Inflammatory Markers in Serum of Spontaneous Rotavirus-Infected Diarrhoetic Calves.

The objective of this study was to assess the efficacy of a microencapsulated probiotic as an adjunct therapy in rotavirus-positive diarrhea of neonatal calves that received supportive treatment or supportive along with microencapsulated probiotic treatment, for 5 days. We examined whether microencapsulated Lactobacillus acidophilus NCDC15 probiotic treatment in rotavirus-infected diarrhoetic calves led to faster resolution of diarrhea, amelioration of zinc-copper imbalance, improved the immunoglobulin A and immunoglobulin G, and decreased the inflammatory markers in serum. Calves with rotavirus-positive diarrhea < 4-week age and fecal scores ≥ 2 were randomly assigned into two groups. The supportive along with microencapsulated probiotic treatment significantly (p < 0.05) increased zinc and immunoglobulin A concentrations and decreased copper, tumor necrosis factor-α, and nitric oxide level in serum on days 3 and 5 from pretreatment values; the immunoglobulin G concentration was elevated (p < 0.05) on day 5. The mean resolution time of abnormal fecal score was 5.3 and 3.3 days in supportive treatment and supportive along with microencapsulated probiotic groups, respectively, in log-rank Mantel-Cox test. The calves in the supportive along with microencapsulated probiotic treatment group had faster resolution of diarrhea than supportive treatment group in Dunn's multiple comparisons test. This study demonstrates that supportive treatment along with microencapsulated probiotic administered to naturally rotavirus-infected diarrhoetic calves at onset of diarrhea led to faster resolution of diarrhea, improved zinc and immunoglobulin levels, and decreased the inflammatory parameters in serum of rotavirus-infected diarrhoetic calves.

Expression and functional role of fibroblast growth factors (FGF) in placenta during different stages of pregnancy in water buffalo (Bubalus bubalis).

The present study documented the expression and functional role of Fibroblast growth factors (FGFs) family and their receptors (Fibroblast growth factor receptor, FGFRs) in placenta (Cotyledon; COT, Caruncle; CAR) during different stages of pregnancy in water buffalo. Samples were collected from Early pregnancy 1 (EP1); Early pregnancy 2 (EP2); Mid pregnancy (MP) and Late pregnancy (LP) while diestrus stage of oestrus cycle (NP) was taken as control. In addition, modulatory role of FGF2 on mRNA expression of von Willebrand factor (vWF), Proliferating cell nuclear antigen (PCNA), steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage (CYP11A1), 3ß-hydroxysteroid dehydrogenase (3ßHSD) and BCL2 Associated X (BAX) were studied in cultured trophoblast cells (TCC), obtained from EP2. Real-time PCR (qPCR), Western blot, and immunohistochemistry were applied to investigate mRNA and protein expressions, and the localization of examined factors whereas, P4 secretion was assessed by RIA. The mRNA and protein expression of FGFs and its receptors were maximum (P < 0.05) during EP (EP1 and EP2) in COT. However, FGFR1 and FGFR4 were upregulated (P < 0.05) during EP2 and MP in COT. Similarly, the mRNA and protein expression of FGFs and its receptors were upregulated (P < 0.05) during all stages of pregnancy in CAR. FGF family members were localized in the cytoplasm of trophoblast cells as well as in fetal blood vessels. At 100 ng/ml dosage, FGF2 stimulated the transcript of vWF maximally (P < 0.05). P4 secretion in trophoblast cells treated with FGF2 was maximum with the highest dose at 72 h. These findings corroborate that FGF acts locally in the trophoblast cells to modulate steroid hormone viz. progesterone synthesis, promote angiogenesis and favors cell survivability indicating that this factor may play an essential role in the regulation of placental formation and function in buffalo.

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.