Intestinal microbiota of broilers submitted to feeding restriction and its relationship to hepatic metabolism and fat mass: Fast‐growing strain

The present study was conducted to verify how feed restriction affects gut microbiota and gene hepatic expression in broiler chickens and how these variables are related to body weight gain. For the experiment, 21‐d‐old Cobb500^TM birds were distributed in a completely randomized experimental design with three treatments: T1. Control (ad libitum—3.176 Mcal/kg ME—metabolizable energy—and 19% CP—crude protein); T2. Energetic restriction (2.224 Mcal/kg ME and 19% CP) from 22 to 42 days with consumption equivalent to control; T3. Quantitative restriction (70% restriction, i.e., restricted broilers ingested only 30% of the quantity consumed by the control group—3.176 Mcal/kg ME and 19% CP) for 7 days, followed by refeeding ad libitum from 28 to 42 days. Ileum and caecum microbiota collections were made at 21, 28 and 42 days of age. Hepatic tissue was collected at 28 and 42 days old for relative gene expression analyses. At 43‐d‐old, body composition was quantified by DXA (Dual‐energy X‐ray Absorptiometry). Both feed restriction programmes decreased Lactobacillus and increased Enterococcus and Enterobacteriaceae counts. No differences were found in the refeeding period. Energetic restriction induced the expression of CPT1‐A (Carnitine palmitoyltransferase 1A) gene, and decreased body fat mass. Quantitative feed restriction increased lipogenic and decreased lipolytic gene expression. In the refeeding period, CPT1‐A gene expression was induced, without changing the broilers body composition. Positive associations were found between BWG (Body Weight Gain) and Lactobacillus and Clostridium cluster IV groups, and negatively associations with Enterobacteriaceae and Enterococcus bacterial groups. In conclusion, differences found in microbiota were similar between the two feed restriction programmes, however, hepatic gene expression differences were only found in quantitative restriction. Higher counts of Lactobacillus and Clostridium cluster IV groups in ileum are likely to be related to better broiler performance and low expression of lipogenic genes.     

Characterization of ileal bacterial microbiota in newly-weaned pigs in response to feeding lincomycin, organic acids or herbal extract

The changes of ileal bacterial microbiota in pigs during the first 2 weeks post-weaning in response to feeding lincomycin, organic acids or herbal extract were characterized using 16S rRNA gene-based PCR-denaturing gradient gel electrophoresis (DGGE) profiling, DNA sequencing, and real-time PCR (QPCR) techniques. Both time post-weaning and the dietary treatments resulted in a shift of the microbiota composition. While time post-weaning mostly influenced Clostridium group, the feed additives increased the population of Lactobacillus and related lactic acid bacteria by ≥ 3-fold.     

Relationships between woodlice (Isopoda: Oniscidea) and microbial density and activity in the field

 Field studies on microbial populations and their activity in soil and litter layers of different sites revealed statistically significant correlations with climatic factors, soil characteristics and the number of terrestrial isopods (Isopoda: Oniscidea). The number of captured Porcellio scaber was negatively correlated with the density of soil microbiota, but positively correlated with both respiration and cellulolytic activity of litter-colonizing microorganisms. By contrast, the number of Oniscus asellus was positively correlated with soil microbial density and respiration, and cellulolytic and respiratory activity of the litter. These interspecific differences were unexpected between species that frequently occur sympatrically, and, thus, probably have similar ecological requirements. Hence, the results suggested species-specific interactions of these species with soil and litter-colonizing microbiota. However, further field studies are required to prove this. Received: 16 December 1998     

尼罗罗非鱼表皮和鳃黏膜共生菌结构特征及其与鱼体健康状况相关关系

黏膜及其表面的共生菌群是鱼类抵御外界不利环境的第一道屏障。为探索养殖罗非鱼表皮和鳃黏膜共生菌菌群的结构特征是否与其健康状况间存在相关关系,本研究运用高通量测序技术,以无乳链球菌腹腔注射攻毒48 h后存活和濒死尼罗罗非鱼为检测对象,检测攻毒前后罗非鱼表皮和鳃黏膜共生菌群落结构差异。结果显示,健康尼罗罗非鱼表皮和鳃黏膜共生菌均存在优势菌群,主要为特吕珀菌属、硫杆菌属、弓形杆菌属、海单胞菌属和弧菌属。人工感染无乳链球菌后存活尼罗罗非鱼表皮和鳃黏膜共生菌菌群与感染前无显著差异;与存活组相比,濒死尼罗罗非鱼的表皮和鳃黏膜共生菌菌群多样性下降,其中弓形杆菌属、假交替单胞菌属、海单胞菌属、假单胞菌属和弧菌属等含量显著下降,链球菌属含量占总菌群的55.30%±1.24%,表明养殖尼罗罗非鱼表皮和鳃黏膜共生菌群落结构可能与其健康状况相关。     

解淀粉芽孢杆菌和胶红酵母复合菌对虹鳟生长性能及胃黏膜、肠黏膜菌群结构的影响

为研究饲料中添加不同水平的复合菌剂(解淀粉芽孢杆菌,Bacillus amyloliquefaciens V4和胶红酵母,Rhodotorula mucilaginosa)对虹鳟(Oncorhynchus mykiss)幼鱼生长及消化道黏膜微生物菌群结构的影响,选用体重为(205.1±4.82)g的虹鳟幼鱼360尾,随机分为4组(每组3个重复,每个重复30尾),分别投喂基础饲料(C0)和3种添加水平为5×10~6/5×10~7 CFU/g(T1),1.5×10~7/1.5×10~8 CFU/g(T3),2.5×10~7/2.5×10~8 CFU/g(T5)的复合菌剂(B.amyloliquefaciens V4/R.mucilaginosa),实验周期42 d。研究结果发现饲料中添加复合益生菌对虹鳟的生长及存活有一定的促进和提高,T1比例的复合益生菌能够显著提高虹鳟的增重率和特定生长率、显著降低饲料系数(P0.05),同时T1和T3比例添加显著降低虹鳟的死亡率(P0.05);对其消化道黏膜细菌群落16S rDNA进行聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)指纹分析,结果表明:虹鳟胃黏膜和肠黏膜上微生物菌群种类存在差异;胃黏膜菌群DGGE图谱中分别检测到35.7±17.0(C0)、37.0±3.5(T1)、36.7±13.6(T3)、26.0±13.2(T5)条谱带,各处理组间谱带数目无显著差异(F=0.500,P=0.692),肠黏膜菌群DGGE图谱显示分别检测到23.3±5.8(C0)、22.3±3.2(T1)、16.7±8.0(T3)、24.7±7.4(T5)条谱带,各处理组间谱带数目也无显著差异(F=0.916,P=0.475);胃黏膜菌群多样性随着益生菌添加量增加,菌群多样性有升高趋势,但是在最高浓度组(T5)多样性降低,肠黏膜菌群多样性随着复合菌剂的添加,多样性指数持续降低,中浓度添加组(T3)多样性最低,但是随着添加浓度升高,呈现恢复和升高趋势(T5);基于所得PCR-DGGE指纹图谱中谱带丰度值数据的UPGMA聚类和PCA排序分析均显示胃黏膜微生物群落与肠黏膜微生物群落结构差异明显,大致分为两个不同的分支,胃黏膜和肠黏膜微生物菌群并没有按照不同处理组而有显著分化。以上结果表明解淀粉芽孢杆菌和胶红酵母复合添加能显著促进虹鳟的生长,提高存活率,外源益生菌添加对虹鳟消化道黏膜上优势菌群能产生一定影响,但并未对胃黏膜及肠黏膜菌群多样性产生显著影响,同时也未显著改变虹鳟肠黏膜微生物菌群结构,高比例添加降低消化道黏膜细菌数量及多样性风险。     

Effect of Bacillus cereus as a water or feed additive on the gut microbiota and immunological parameters of Nile tilapia

We evaluated the effects of Bacillus cereus, as an additive in water and feed, on the gut microbiota and immunological parameters of Nile tilapia (Oreochromis niloticus) fingerlings. Experiments were performed in tanks and net cages respectively. Experiment 1: Tilapia were housed in tanks for 42 days, and B. cereus was added to the water at 1.0 × 10⁴ cfu mL^?1 (Treatment 1) and 1.0 ×10⁵ cfu mL^?1 (Treatment 2) weekly. For the control, no probiotic was added. Experiment 2: Tilapia were housed in cages for 42 days, and the feed was supplemented with B. cereus at 1.0 × 10⁷ cfu g^?1 (Treatment 1) and 1.0 × 10⁸ cfu g^?1 (Treatment 2) weekly. For the control, no probiotic was added. Each treatment contained three replicates, with 50 male tilapias per replicate. The fish from the probiotic treatments in both tank and cage experiments had significantly higher serum lysozyme and peroxidase activities than the control. In the cage experiment, alkaline phosphatase and total superoxide dismutase activities in tilapia were significantly higher in probiotic treatments compared with the control. The results of polymerase chain reaction‐denaturing gradient gel electrophoresis showed that B. cereus supplementation in the feed and water affected the autochthonous gut bacteria community of tilapia and stimulated various potentially beneficial bacteria. Therefore, B. cereus, as a water or feed additive, could enhance the immune status and affect the gut microbiota of tilapia. Bacillus cereus was more effective as a feed supplement rather than a water additive for enhancing the immune status of tilapia.     

Two biomass preparation methods provide insights into studying microbial communities of intestinal mucosa in grass carp (Ctenopharyngodon idellus)

Animal digestive tract is habitat for a large number of autochthonous microbiota, which play central roles in multiple biological and physiological processes of the host. In this study, two different micro‐biomass preparation methods were employed to evaluate the diversity of intestinal mucosa‐associated microbiota in grass carp (Ctenopharyngodon idellus). Genomic DNAs were isolated either directly from intestinal mucosal samples (group A), or from micro‐biomass after microbial dissociation (group B). Community richness, diversity and evenness indices were all higher in group B, but differences were not statistically significant (P = 0.97, P = 0.33, P = 0.34 respectively). Furthermore, group B samples exhibited an increased ratio of bacterial DNA in comparison with group A samples, but the difference was also not statistically significant (P = 0.74). In addition, there were no statistically significant differences between the two groups (P > 0.05) at the taxonomic level. Our results support previous findings that there exists a great abundance of the intestinal mucosa‐adherent microbiota in the grass carp; among these, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Spirochaetes and Fusobacteria were the most common phyla. Within these microbiota, Paenibacillus, Bacteroides, Bacillus and Cetobacterium genera comprise the majority of the community, implicating their functional importance (e.g. as probiotics) to their host. Our results contribute towards a better understanding of the intestinal microbial profile of grass carp. Both micro‐biomass preparation techniques proved to be feasible for studying mucosa‐adherent microbiota of grass carp; however, the second method (group B) provides a protocol that is somewhat more effective than the first method (group A).     

Impacts of diet on hindgut microbiota and short‐chain fatty acids in grass carp (Ctenopharyngodon idellus)

Diet is known to influence intestinal microbiota in fish, but the specifics of these impacts are still poorly understood. Different protein/fibre ratio diets may result in differing structures and activities of gut microbiota. We examined the hindgut microbiome of grass carp (Ctenopharyngodon idellus) fed three different diets: fish meal (FM, high protein – low fibre), Sudan grass (SG, high fibre – low protein) and compound feed (CF, intermediate). Microbial profiles of fish fed on FM were significantly different from profiles of fish fed CF and SG (F = 18.85, p < .01). Cetobacterium, known to be positively associated with protein digestion, was the dominant microbial group in FM samples (approximately 75.7%), while Lachnospiraceae and Erysipelotrichaceae, thought to be involved in fermentation of plant polysaccharides, were dominant in CF and SG samples (46.8% and 42.9% respectively). Network analyses indicated that the abundance of Lachnospiraceae and Erysipelotrichaceae was in a significantly positive correlation (r = .895, p = .001). Short‐chain fatty acid (SCFA) levels may indicate that the digestibility of diet by microbiota in the grass carp gut decreased from FM to SG (FM>CF>SG). Overall low SCFA levels indicate that hindgut fermentation probably provides a low proportion of energy requirements in grass carp.