Diversity and activity of cellulolytic bacteria,isolated from the gut contents of grass carp (Ctenopharyngodon idellus) (Valenciennes) fed on Sudan grass (Sorghum sudanense) or artificial feedstuffs

Herbivorous grass carp (Ctenopharyngodon idellus) has a powerful capability to digest cellulose from aquatic plants, depending on the cellulase complex produced by the cellulolytic bacterial community in the gastrointestinal (GI) tract. However, it remains uncertain which bacteria taxa may actively participate in the digestion of food fibre. In this study, a total of 499 cellulolytic bacteria from the gut content of grass carp fed on Sudan grass (242 strains) and artificial feedstuffs (257 strains) were randomly isolated and characterized using carboxymethyl‐cellulose, microcrystalline cellulose and cellobiose agar media. The results showed that more than half of the isolates were capable of degrading carboxymethyl‐cellulose and cellobiose, while the remaining isolates were restricted to microcrystalline cellulose decomposition, exclusively. The cellulolytic bacterial community was dominated by Aeromonas, followed by Enterobacter, Enterococcus, Citrobacter, Bacillus, Raoultella, Klebsiella, Hydrotalea, Pseudomonas, Brevibacillus and some unclassified bacteria, as revealed by 16S rDNA sequence analysis. Notably, grass carp fed on grass with high‐fibre content harboured a higher diversity of cellulolytic bacteria than the ones fed on low‐fibre feedstuffs. Our results provided evidence for a positive correlation between the content of food fibre and the diversity of cellulolytic bacteria in grass carp intestines. Thus, improving growth conditions and cellulase activities for GI cellulolytic microorganisms in grass carp intestines are critical for effective utilization of feedstuffs containing high fibre levels.     

不同纤维源对成年鹅肠道组织形态变化的影响

本试验旨在研究不同纤维源对成年扬州鹅肠道组织形态变化的影响。选用18只健康、体重相近的25周龄扬州鹅公鹅,随机分为3组,即:稻壳粉组、苜蓿草粉组、羊草粉组,每组6只。试验期28 d,试验期末将试验鹅处死并留取各肠段组织,称重并观察肠道组织形态。结果表明:①不同纤维源对十二指肠和空肠长度影响差异不显著(P>0.05),稻壳草粉组回肠长度显著高于苜蓿草粉组与羊草粉组(P<0.05),苜蓿草粉组与羊草粉组差异不显著(P>0.05)。②稻壳粉组十二指肠绒毛高度、绒毛宽度、隐窝深度、肌层厚度均显著高于苜蓿草粉组与羊草粉组(P<0.05),其中苜蓿草粉组隐窝深度、V/C与羊草粉组差异显著(P<0.05);稻壳粉组与羊草粉组空肠绒毛高度显著高于苜蓿草粉组(P<0.05),隐窝深度显著低于苜蓿草粉组(P<0.05),稻壳粉组与羊草粉组间均差异不显著(P>0.05),稻壳粉组肌层厚度显著高于苜蓿草粉组(P<0.05);羊草粉组回肠绒毛高度显著高于苜蓿草粉组与稻壳粉组(P<0.05),3组间绒毛宽度差异显著(P<0.05),且以稻壳粉组最大,羊草粉组V/C显著高于苜蓿草粉组和稻壳粉组(P<0.05),羊草粉组肌层厚度显著低于苜蓿草粉组与稻壳草粉组(P<0.05)。由此可知,不同纤维源可显著影响25周龄扬州鹅肠道组织形态。     

肠道微生物宏基因组文库构建及酯解酶基因筛选

利用宏基因组学方法,以人肠道微生物样品为原材料,构建了1个约30 000个克隆的fosmid文库.以三丁酸甘油酯为底物,通过功能筛选,获得1个酯解酶阳性克隆.对该阳性克隆构建亚克隆,挑选具有酯解酶活性的阳性亚克隆进行测序分析,最终获得1个肠道微生物来源的酯解酶基因(GenBank登录号:JQ972699).结果表明,文库克隆的平均插入片段约为40 kb,没有重复插入片段克隆.获得的酯解酶基因推演蛋白与Pyramidobacter piscolens W5455的patatin样磷脂酶同源性最高,氨基酸一致性为95％.生物信息学分析结果表明该基因可能通过Vd型分泌方式进行分泌并发挥功能.本研究是通过构建人肠道微生物宏基因组大片段文库并结合重组子功能筛选获得酯解酶的首次报道,可为食品工业提供新的酯解酶来源和筛选方法.     

Highly metastatic ovarian yolk sac carcinoma in a rat

We investigated a highly metastatic ovarian yolk sac carcinoma in a 52-week-old female Crl:CD(SD) rat. Macroscopically, the present case had severe ascites, bilateral ovarian masses and numerous nodules in the abdominal and thoracic cavities. Histopathologically, these masses and nodules were generally composed of two types of cells mimicking a parietal and visceral yolk sac. The parietal cells were round to polygonal, contained eosinophilic droplets and were arranged in nests and cords in the eosinophilic matrix. Both the intracytoplasmic droplets and the matrix were stained positively with PAS. The visceral cells were cylindriform, and proliferated in papillary and tubular patterns and occasionally formed Shiller-Duval body-like structures. In the dissemination sites, the neoplastic cells proliferated on the surface of the various tissues and often infiltrated into deeper parts of the tissues. Immunohistochemically, both neoplastic cells were positive for α-fetoprotein and keratin, and the eosinophilic matrix was positive for laminin. Ultrastructurally, the parietal cells had dilated rough endoplasmic reticulums, which were filled with electron-lucent laminated structures. The visceral cells had poorly to moderately developed intracytoplasmic organelles and were interconnected with desmosomes. Taken together, the present tumor was diagnosed as yolk sac carcinoma arising from the ovary and was characterized by not only high metastasis but also invasive infiltration with biphasic proliferation of the parietal and visceral cells.     

Effects of three major protein sources on performance,gut morphology and fermentation characteristics in broilers

ABSTRACT

1. This study determined the effects of three protein sources (PS), each at two digestibility crude protein (DCP) levels, on performance, gut morphology and fermentation characteristics in the hindgut of broilers.

2. It was hypothesised that broilers fed ingredients high in indigestible CP, i.e. rapeseed meal (RSM) or maize gluten (MG), could potentially cause reduced growth, impaired gut health, and more protein fermentation products in caecal digesta. Increasing the DCP level in each of the indigestible CP diets may compensate for these detrimental effects.

3. In total, 288 one-d-old male Ross 308 broilers were used in a completely randomised 3 × 2 factorial design, with six replicate pens per treatment. Three PS: soybean meal (SBM), rapeseed meal (RSM) or maize gluten (MG), and two DCP levels: 15.8 and 17.2% were used.

4. Broilers fed SBM had increased feed intake and BWG and improved FCR compared with those fed RSM and MG diets. Broilers fed high DCP had better performance compared with those on low DCP. No significant effects of PS or DCP level were found on gastrointestinal tract development, caecal ammonia or volatile fatty acid concentrations.

5. Broilers fed SBM had longer villi, smaller crypts and increased villus height to crypt depth ratio compared with those fed RSM and MG diets. Broilers fed RSM diet had a lower caecal pH, and had 16.5% and 14.9% more branched chain fatty acid contents in caecal digesta compared with those fed SBM and MG diets, respectively, indicating more proteolytic fermentation.

6. Replacing SBM by RSM and MG negatively affected growth performance and gut morphology. Hindgut protein fermentation was substantially increased in RSM fed birds.

7. To a certain extent, retarded growth performance in RSM and MG fed birds could be counterbalanced by increasing the dietary level of digestible CP.     

不同粪菌悬液处理方式对猪粪菌移植过程中细菌活性及活菌组成的影响

旨在探究不同粪菌液处理方法对其菌群活性及存活菌群物种组成的影响,为猪粪菌移植（FMT）的优化应用提供参考。本研究共设置7个试验组制备粪菌液：静置制备新鲜粪菌液组（FS）、静置制备粪菌液于-80℃冷冻1周组（FFS）、离心制备新鲜粪菌液组（FC）、离心制备粪菌液于-80℃冷冻1周组（FFC）、液氮冷冻粪便24 h静置制备粪菌液组（NS）、甘油+液氮冷冻粪便24 h静置制备粪菌液组（GNS）和液氮冷冻粪便24 h静置制备粪菌液于-80℃冷冻1周组（NFS）,在37℃厌氧培养箱中平板培养48 h后,进行菌落计数。从FS、FFS、FC和FFC 4组中根据菌落形态随机挑取87个单菌落纯化后进行16S rRNA基因全长测序。结果表明,静置和离心两种方法制备新鲜粪菌液时,粪菌活性存在显著差异,离心组显著低于静置组（P<0.05）。但冷冻粪便使用静置和离心两种方法制备的菌悬液,其粪菌活性无显著差异（P>0.05）。无论是静置还是离心方法制备粪菌液,新鲜和冷冻两种不同处理方式下的粪菌活性均无显著差异（P>0.05）,且对活菌的组成也无显著影响。使用静置法制备粪菌液时,FS、NS、GNS和NFS组间粪菌活性无显著差异（P>0.05）。综上表明,静置方法制备粪菌液时,超低温冷冻保存对粪菌活性及其活菌组成均无显著影响,这为大样本、长距离猪粪菌移植技术的应用提供了重要试验参考。     

Denaturing gradient gel electrophoresis analysis of gut bacterial community for the Japanese earthworms

The gut bacterial community structure for Pheretima hilgendorfi and P. heteropoda (Family Megascolecidae), and Allolobophora japonica (Family Lumbricidae) collected from agricultural grasslands in Japan was analyzed by denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16S rRNA gene fragments (PCR-DGGE) and compared with those in the surrounding soils. Denaturing gradient gel electrophoresis (DGGE) profiles indicated that each earthworm species had their own specific bacterial communities, and multidimentional scaling analysis grouped the DGGE profiles into three groups: gut samples from P. hilgendorfi and P. heteropoda, gut samples from A. japonica and samples from the surrounding soils. Nine dominant bands were identified by their direct sequencing and cloning. Major three bands from P. hilgendorfi and P. heteropoda were closely related to Bacillus species belonging to the phylum Firmicutes. Major four and two bands from A. japonica were closely related to the phyla Proteobacteria and Bacteroidetes, respectively.     

Potential for endozoochorous seed dispersal by sheep and goats: Risk of weed seed transport via animal faeces

Endozoochory is known as an important mechanism for the spread of weeds. We carried out experiments to assess the fate of seeds of several weed species (Convolvulus arvensis, Cuscuta campestris, Rumex crispus, Hordeum spontaneum and Sorghum halepense) after passing through the gut of sheep and goat. Eighteen animals of both sheep and goat received diet mixed with seeds of the weed species or control with only wheat bran (five weed species + control × three replications). Results showed that a higher proportion of seeds were missing after passage through the sheep gut than in goats. In goats, a greater proportion of seeds were dead after passage, but the number of seeds collected from dung was also greater. Weed species differed, with the highest seed recovery and viability in Cuscuta campestris. Based on time of seed passages through the animal gut estimated for the different weed species, we recommend that sheep should be kept in a corral for 96 hr to minimise seed transportation via their faeces. For goats, if R. crispus and C. arvensis seeds could be excluded from the diet, then maintaining them for 96 hr in an animal stall would ensure little seed transportation via dung, but we found R. crispus and C. arvensis seeds to be present and viable in goat dung even 120 hr after feeding. Very large numbers of viable seeds can be found in goat and sheep dung, so the use of rotted manure is highly recommended to avoid transportation of viable seeds via manure fertilisers.     

Ontogeny of digestive tract functionality in Japanese flounder, Paralichthys olivaceus studied by in vivo microinjection: pH and assimilation of free amino acids

To specifically study the functional ontogeny of the digestive tract, larvae of Japanese flounder at various developmental stages were injected with liquid solutions using tube feeding in vivo. The survival in the stages tested was on average 93%. The injected solutions were almost completely transferred from the presumptive stomach, or stomach to the midgut within 10 min of injection. The passage to the hindgut in some cases started 10 min after injection and over 90% of the solution had passed from the midgut to the hindgut after 1 h. In most cases the hindgut seemed to be completely empty after 3 h.Two different mixtures of pH indicators with sensitivities in the alkaline (7.5 to 9) and acid (4 to 6) ranges respectively were used for assessment of pH in the various gut segments. The pH of the stomach remained alkaline during the larval period, but had fallen close to 4 during late metamorphosis, an indication of active HCl secretion and progressive stomach differentiation. In mid and late metamorphosing fish a rapid colour change in the pH indicator was observed once it had passed the pyloric sphincter. This demonstrates that there was also active secretion of alkaline fluid, most likely HCO₃ ^–, from the pancreas into the pylorus lumen.A single injection of liquid solution of ¹⁴C-FAA showed that assimilation of FAA was high in all stages tested (79.5 ± 7.1%; SD; n=91). The presently reported data for Japanese flounder support earlier studies that FAA are absorbed with a high efficiency in the early stages of marine fish.     

益生菌对类风湿关节炎的改善作用研究进展

类风湿关节炎是一种威胁全球人类健康的自身免疫性疾病,肠道菌群的平衡与类风湿关节炎密切相关。由于益生菌在改善肠道菌群紊乱、恢复正常肠道菌群中发挥重要作用,因此给益生菌治疗类风湿关节炎提供了可能。本文综述类风湿关节炎病因、机制及患者肠道菌群的变化,总结近年来益生菌在治疗类风湿关节炎中的研究现状,为进一步临床试验和今后的应用提供依据。