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IntroductionGrowth-retarded lambs would reduce the economic incomes of sheep farming. Nutritional interventions are supposed to promote gastrointestinal health and the compensatory growth of growth-retarded lambs. This study evaluated the effects of probiotic supplementation on the growth performance, plasma characteristics and ruminal bacterial community of growth-retarded lambs.MethodsTwenty-four 50-days old male Hu lambs, including 8 healthy lambs (13.2 ± 1.17 kg) and 16 growth-retarded lambs (9.46 ± 0.81 kg), were used in this study. The 8 healthy lambs were fed the basal diet and considered the positive control (GN), and the other 16 growth-retarded lambs were randomly assigned into 2 groups (basal diet without probiotic [negative control, GR] and basal diet supplementation with 1 g/kg concentrate feed probiotic [GRP]), with each group having 4 replicate pens. The feeding trial lasted for 60 days with 7 days for adaptation.ResultsThe results showed that dietary supplementation with probiotic increased (p < 0.05) the average daily gain and dry matter intake of growth-retarded lambs. For growth-retarded lambs, supplementation with probiotic increased (p < 0.05) the activities of superoxide dismutase and glutathione peroxidase, as well as the concentrations of growth hormone and immunoglobulin G. Furthermore, the highest (p < 0.05) concentrations of interleukin-6, interferon-gamma and tumor necrosis factor alpha were observed in the GR group. The concentrations of total volatile fatty acids and acetate in growth-retarded lambs were increased by probiotic supplementation (p < 0.05). The relative abundances of Ruminococcus, Succiniclasticum and Acidaminococcus were lower (p < 0.05) in growth-retarded lambs. However, probiotic supplementation increased (p < 0.05) the relative abundances of these three genera.DiscussionThese results indicate that dietary supplementation with probiotic are promising strategies for improving the growth performance of growth-retarded lambs by enhancing immunity and altering the ruminal microbiota.

Weaning plays an important role in many animal processes, including the development of the rumen microbiota in ruminants. Attaining a better understanding of the development of the rumen microbial community at different weaning stages can aid the identification of the optimal weaning age. We investigated the effects of weaning age on ruminal bacterial and archaeal communities in Hu lambs. Thirty male Hu lambs were randomly assigned to two weaning-age groups: a group weaned at 30 days of age (W30) and a group weaned at 45 days of age (W45), with each group having five replicate pens. On the weaning day (day 30 for W30 and day 45 for W45) and at 5 days postweaning [day 35 for W30 (PW30) and day 50 for W45 (PW45)], one lamb from each replicate was randomly selected and sacrificed. Rumen contents were collected to examine the ruminal microbiota. Compared to W30, PW30 had a decreased relative abundance of Bacteroidetes. At genus level, the extended milk replacer feeding (W45 vs. W30) increased the relative abundance of Ruminococcus while decreased that of Prevotella and Dialister. Compared to W30, PW30 exhibited decreased relative abundances of Prevotella, Dialister and Bacteroides but an increased unclassified Coriobacteriaceae. No significant difference was noted in the detected archaeal taxa among the animals. The function “biosynthesis of secondary metabolites” was less predominant in PW30 than in W30, whereas the opposite held true for “metabolism of cofactors and vitamins.” Some bacterial genera were significantly correlated with rumen volatile fatty acid (VFA) concentration or other animal measures, including negative correlations between ruminal VFA concentration and unclassified Mogibacteriaceae and unclassified Veillonellaceae; positive correlations of ruminal papillae length with Fibrobacter and unclassified Lachnospiraceae, but negative correlations with Mitsuokella and Succiniclasticum; and negative correlations between plasma D-lactate concentration and Prevotella, unclassified Paraprevotellaceae, and Desulfovibrio. Our results revealed that the ruminal bacterial community underwent larger changes over time in lambs weaned at 30 days of age than in lambs weaned half a month later. Thus, extending milk replacer feeding to 45 days weaning was recommended from the perspective of the rumen microbial community in the Hu lamb industry.

Defaunation studies have shown that rumen protozoa are one of the main causes of low nitrogen utilization efficiency due to their bacterivory and subsequent intraruminal cycling of microbial protein in ruminants. In genomic and transcriptomic studies, we found that rumen protozoa expressed lysozymes and peptidases at high levels. We hypothesized that specific inhibition of lysozyme and peptidases could reduce the activity and growth of rumen protozoa, which can decrease their predation of microbes and proteolysis and subsequent ammoniagenesis by rumen microbiota. To test the above hypothesis, we evaluated three specific inhibitors: imidazole (IMI), a lysozyme inhibitor; phenylmethylsulphonyl fluoride (PMSF), a serine protease inhibitor; and iodoacetamide (IOD), a cysteine protease inhibitor; both individually and in combinations, with sodium dodecyl sulfate (SDS) as a positive control. Rumen fluid was collected from two Jersey dairy cows fed either a concentrate-based dairy ration or only alfalfa hay. Each protozoa-enriched rumen fluid was incubated for 24 h with or without the aforementioned inhibitors and fed a mixture of ground wheat grain, alfalfa, and grass hays to support microbial growth. Live protozoa cells were morphologically identified and counted simultaneously at 3, 6, 12, and 24 h of incubation. Fermentation characteristics and prokaryotic composition were determined and compared at the end of the incubation. Except for IOD, all the inhibitors reduced all the nine protozoal genera identified, but to different extents, in a time-dependent manner. IOD was the least inhibitory to protozoa, but it lowered ammoniagenesis the most while not decreasing feed digestibility or concentration of volatile fatty acids (VFA). ANCOM analysis identified loss of Fibrobacter and overgrowth of Treponema, Streptococcus, and Succinivibrio in several inhibitor treatments. Functional prediction (from 16S rRNA gene amplicon sequences) using the CowPI database showed that the inhibitors decreased the relative abundance of the genes encoding amino acid metabolism, especially peptidases, and lysosome in the rumen microbiota. Overall, inhibition of protozoa resulted in alteration of prokaryotic microbiota and in vitro fermentation, and peptidases, especially cysteine-peptidase, may be targeted to improve nitrogen utilization in ruminants.