Effect of hay type on cecal and fecal microbiome and fermentation parameters in horses

The effect of hay type on the microbiome of the equine gastrointestinal tract is relatively unexplored. Our objective was to characterize the cecal and fecal microbiome of mature horses consuming alfalfa or Smooth Bromegrass (brome) hay. Six cecally cannulated horses were used in a split-plot design run as a crossover in two periods. The whole plot treatment was ad libitum access to brome or alfalfa hay fed over two 21-d acclimation periods with subplots of sampling location (cecum and rectum) and sampling hour. Each acclimation period was followed by a 24-h collection period where cecal and fecal samples were collected every 3 h for analysis of pH and volatile fatty acids (VFA). Fecal and cecal samples were pooled and sent to a commercial lab (MR DNA, Shallowater, TX) for the amplification of the V4 region of the 16S rRNA gene and sequenced using Illumina HiSeq. The main effects of hay on VFA, pH, and taxonomic abundances were analyzed using the MIXED procedure of SAS 9.4 with fixed effects of hay, hour, location, period, and all possible interactions and random effect of horse. Alpha and beta diversities were analyzed using the R Dame package. Horses fed alfalfa had greater fecal than cecal pH (P ≤ 0.05), whereas horses fed brome had greater cecal than fecal pH (P ≤ 0.05). Regardless of hay type, total VFA concentrations were greater (P ≤ 0.05) in the cecum than in feces, and alfalfa resulted in greater (P ≤ 0.05) VFA concentrations than brome in both sampling locations. Alpha diversity was greater (P ≤ 0.05) in fecal compared with cecal samples. Microbial community structure within each sampling location and hay type differed from one another (P ≤ 0.05). Bacteroidetes were greater (P ≤ 0.05) in the cecum compared with the rectum, regardless of hay type. Firmicutes and Firmicutes:Bacteroidetes were greater (P ≤ 0.05) in the feces compared with cecal samples of alfalfa-fed horses. In all, fermentation parameters and bacterial abundances were impacted by hay type and sampling location in the hindgut.     

猪肠道细菌培养组学研究进展

猪肠道菌群是由所有定殖在猪肠道里的大量细菌、病毒、真菌和古菌等构成的集合。已有研究表明,很多疾病以及猪重要经济性状都与肠道菌群有关。目前,肠道菌群研究使用较多的技术是16S rRNA基因测序和宏基因组测序,但这些技术并不能了解具体菌株的实际功能和生理特性。肠道细菌的分离培养具有特殊重要的意义。近几年来,肠道细菌的培养取得了重要进展,基于培养条件的多样性分离培养出了更多的肠道细菌类别,这对促进肠道菌群在菌株水平的研究以及推广应用有重要意义。本文主要从猪肠道菌群组成结构、培养组学发展、猪肠道细菌培养组学的研究现状等方面进行论述和展望,为后续猪肠道菌群菌株的功能和影响表型的机制研究提供参考借鉴。     

玉米花丝多糖的提取制备与抑菌作用研究

本研究以玉米花丝为原料提取多糖。选取花丝类型、花丝和水的比例、温度和时间4个因素,采用L9(34)正交设计,两次重复。结果表明:玉米花丝多糖提取的最佳方法为:采用老花丝,花丝和水比例为1:6,60℃水浴2.5 h,提取率为3.65%。通过浓缩、离心、沉淀等过程,并去除蛋白质、色素等杂质,得到玉米花丝粗多糖。将粗多糖进行甲醇分级纯化,得到4种纯化多糖。再将4种多糖各配制成6种浓度,对活化后的8种细菌和4种真菌进行抑菌试验,测量抑菌圈直径,比较抑菌效果。结果表明:玉米花丝多糖对细菌有明显的抑制作用,对真菌也有一定的抑制作用,抑菌圈直径为11.5 mm。其中,以40%甲醇浓度,4000 r/min和45%甲醇浓度,5000 r/min进行离心分离的多糖对细菌和真菌的抑制效果较好。     

Cashew nut shell liquid potentially mitigates methane emission from the feces of Thai native ruminant livestock by modifying fecal microbiota

The methane-mitigating potency of cashew nutshell liquid (CNSL) was evaluated by investigating gas production from batch cultures using feces from Thai native ruminants that had been incubated for different periods. Feces was obtained from four Thai native cattle and four swamp buffaloes reared under practical feeding conditions at the Kasetsart University farm, Thailand. Fecal slurry from the same farm was also included in the analysis. CNSL addition successfully suppressed the methane production potential of feces from both ruminants by shifting short chain fatty acid profiles towards propionate production. Methane mitigation continued for almost 150 days, although the degree of mitigation was more apparent from Day 0 to Day 30. Bacterial and archaeal community shifts with CNSL addition were observed in feces from both ruminants; specifically, Bacteroides increased, whereas Lachnospiraceae and Ruminococcaceae decreased in feces to which CNSL was added. Fecal slurry did not show marked changes in gas production with CNSL addition. The findings showed that the addition of CNSL to the feces of ruminants native to the Southeast Asian region can suppress methane emission. Because CNSL can be easily obtained as a byproduct of the local cashew industry in this region, its on-site application might be ideal.     

Growth and morphologic response of rumen methanogenic archaea and bacteria to cashew nut shell liquid and its alkylphenol components

The growth and morphology of rumen methanogenic archaea (15 strains of 10 species in 5 genera, including 7 strains newly isolated in the present study) and bacteria (14 species in 12 genera) were investigated using unsupplemented in vitro pure cultures and cultures supplemented with cashew nut shell liquid (CNSL) and its phenolic compound components, anti-methanogenic agents for ruminant animals. Growth of most of the methanogens tested was inhibited by CNSL and alkylphenols at different concentrations ranging from 1.56 to 12.5 μg/ml. Of the alkylphenols tested, anacardic acid exhibited the most potent growth inhibition. Three gram-negative bacterial species involved in propionate production were resistant to CNSL and alkylphenols (>50 μg/ml). All the methanogens and bacteria that were sensitive to CNSL and alkylphenols exhibited altered morphology; disruption of the cell surface was notable, possibly due to surfactant activity of the tested materials. Cells division was inhibited in some organisms, with cell elongation and unclear septum formation observed. These results indicate that CNSL and alkylphenols, particularly anacardic acid, inhibit both rumen bacteria and methanogens in a selective manner, which could help mitigate rumen methane generation.     

Isolation and Identification of Pathogenic Bacteria Causing Dairy Cow Mastitis in Liaoning and Drug Sensibility Test of Pathogenic E.coli

In order to understand the main bacterial pathogen species causing dairy cow mastitis in Liaoning, as well as the characteristics of drug sensitivity of the pathogenic E.coli,the milk samples from 75 dairy cows with clinical manifestations for mastitis in certain large-scale dairy farm in Liaoning were collected.The bacteria in milk were cultured and isolated with biochemical methods and in vitro drug sensitivity tests were processed with the isolated E.coli strains.The results showed that the main bacterial pathogen for dairy cow mastitis were E.coli（separation rate 58.7%),S.aureus（64.0%）and S.agalactiae（54.7%),and multiple infection including double and triple infection were identified.The drug sensitivity tests on the isolated E.coli indicated that the E.coli isolates were highly resistant to sulfonamides（resistance rate>85%）and chloramphenicol（resistance rate>30%),and they were relatively low resistant to ampicillin（9.5%),ciprofloxacin（9.5%),ceftiofur（7.1%）and ofloxacin（4.8%).The results was able to provide reliable theoretical basis for prevention and control of dairy cow mastitis in Liaoning area.     

The effect of lactic acid bacterial starter culture and chemical additives on wilted rice straw silage

Lactic acid bacteria (LAB) are suitable for rice straw silage fermentation, but have been studied rarely, and rice straw as raw material for ensiling is difficult because of its disadvantages, such as low nutrition for microbial activities and low abundances of natural populations of LAB. So we investigated the effect of application of LAB and chemical additives on the fermentation quality and microbial community of wilted rice straw silage. Treatment with chemical additives increased the concentrations of crude protein (CP), water soluble carbohydrate (WSC), acetic acid and lactic acid, reduced the concentrations of acid detergent fiber (ADF) and neutral detergent fiber (NDF), but did not effectively inhibit the growth of spoilage organisms. Inoculation with LABs did not improve the nutritional value of the silage because of poor growth of LABs in wilted rice straw. Inoculation with LAB and addition of chemical materials improved the quality of silage similar to the effects of addition of chemical materials alone. Growth of aerobic and facultatively anaerobic bacteria was inhibited by this mixed treatment and the LAB gradually dominated the microbial community. In summary, the fermentation quality of wilted rice straw silage had improved by addition of LAB and chemical materials.     

Characterization and application of lactic acid bacteria for tropical silage preparation

Strains TH 14, TH 21 and TH 64 were isolated from tropical silages, namely corn stover, sugar cane top and rice straw, respectively, prepared in Thailand. These strains were selected by low pH growth range and high lactic acid‐producing ability, similar to some commercial inoculants. Based on the analysis of 16S ribosomal RNA gene sequence and DNA‐DNA relatedness, strain TH 14 was identified as Lactobacillus casei, and strains TH 21 and TH 64 were identified as L. plantarum. Strains TH 14, TH 21, TH 64 and two commercial inoculants, CH (L. plantarum) and SN (L. rhamnosus), were used as additives to fresh and wilted purple Guinea and sorghum silages prepared using a small‐scale fermentation method. The number of epiphytic lactic acid bacteria (LAB) in the forages before ensilage was relatively low but the numbers of coliform and aerobic bacteria were higher. Sorghum silages at 30 days of fermentation were all well preserved with low pH (3.56) and high lactic acid production (72.86 g/kg dry matter). Purple Guinea silage inoculated with LAB exhibited reduced count levels of aerobic and coliform bacteria, lower pH, butyric acid and ammonia nitrogen and increased lactic acid concentration, compared with the control. Strain TH 14 more effectively improved lactic acid production compared with inoculants and other strains. © 2016 Japanese Society of Animal Science     

Rearing conditions affected responses of weaned pigs to organic acids showing a positive effect on digestibility,microflora and immunity

Three experiments were conducted to assess the response of weaned pigs to organic acid SF3, which contains 34% calcium formate, 16% calcium lactate, 7% citric acid and 13% medium chain fatty acids. Dietary treatments had no effect on growth performance of piglets (21‐day weaning) fed the commercial prestart diet for 1 week before receiving the experimental diets supplemented with SF3 at 0, 3 or 5 g/kg diet (Exp. 1), whereas diarrhea frequency averaged across a week was decreased by SF3 supplementation (5 g/kg diet) in piglets fed the experimental diets immediately after weaning (Exp. 2). In Exp. 3, piglets (28‐day weaning) were fed the control (containing pure colistin sulfate and enramycin, respectively, at 20 mg/kg diet) for 1 week and then were fed the control or SF3‐supplemented (5 g/kg diet) diet for 2 weeks. The SF3‐fed piglets had greater apparent ileal digestibility of calcium and dry matter, while also demonstrating greater overall gross energy, up‐regulated jejunal expression of sodium‐glucose cotransporter‐1 and transforming growth factor‐β, down‐regulated jejunal expression of tumor necrosis factor (TNF)‐α, higher ileal Lactobacillus, with lower total bacteria content, lower plasma TNF‐α but higher IgG levels than the control‐fed piglets. Collectively, SF3 consumption improved diarrhea resistance of weaned pigs by improving nutrient digestibility, piglet immunity and intestinal bacteria profile. © 2016 Japanese Society of Animal Science