虎斑颈槽蛇颈腺的解剖观察研究

虎斑颈槽蛇颈腺的解剖观察研究吴卯斌（安徽省黄山市屯溪毒蛇养殖研究所２４５０５１）关键词虎斑颈槽蛇，颈腺，解剖我所多年从事毒蛇的人工养殖研究工作，在进行蛇类饲养，观察过程中，一次偶尔触怒虎斑颈槽蛇（Ｒｈａｂｄｏｐｈｉｓｔｉｇｒｉｎｕｓ）［又名虎斑游蛇］...     

一种有毒的游蛇——虎斑颈槽蛇

虎斑颈槽蛇是中国数量较多、分布最广的蛇类之一,但一直被当作无毒蛇而忽视。为了使人们对虎斑颈槽蛇有一个全面、正确的认识,也为以后的相关研究奠定一定的理论基础,从虎斑颈槽蛇的分布,形态,生活习性以及毒腺的研究等方面加以综述。     

八种蛇(蛇亚目:游蛇科:颈槽蛇属)的半阴茎形态比较

蛇类半阴茎形态在研究蛇类的形态进化、分类和系统等方面具有重要的作用。对分布于我国境内的颈槽蛇属八种蛇(虎斑颈槽蛇、海南颈槽蛇、颈槽蛇、九龙颈槽蛇、缅甸颈槽蛇、黑纹颈槽蛇、喜山颈槽蛇和红脖颈槽蛇)的半阴茎进行详细描述和比较,并以此探讨部分物种的有效性和推断它们之间的进化及系统关系。     

虎斑颈槽蛇中国大陆亚种颈背腺毒液的研究

虎斑颈槽蛇中国大陆亚种（Rhabdophis tigrinus lateralis）具有已知蛇类中罕见的颈背腺及其毒液。肉眼观察发现腺体呈珠球状,8～13对腺体呈平行排列,腺体间隔分布;组织切片观察发现,腺体上无开口,无合成毒液的细胞器;实验表明毒液有神经毒性,对小白鼠的半致死量（LD50）为97．99mg／kg,SDS-PAGE电泳呈现4条蛋白条带。虎斑颈槽蛇中国大陆亚种颈背腺毒液很可能作为一种特殊辅助防御系统,增强DuvemoY腺毒液的防御作用。     

虎斑颈槽蛇毒液伤眼1例报告

<正>我区地处皖南山区,气候温和,自然条件适宜蛇类生长,蛇类种群繁多。其中以虎斑颈槽蛇分布较广,在我区黄山、青阳、祁门、东至等地均为优势种之一,主要生活于山区、丘陵、平原,常出没玉米地、路边、菜地、水沟边及近水、潮湿的草地等。曾有文献报道,该蛇颈部可分泌毒液,也有临床报道该蛇咬伤人案例发生[1]。我院近年来每年收治毒蛇咬伤患者近100例,但唯一收治1例虎斑颈槽蛇毒液致伤眼患者,现将结果报告如下。     

基于16S rRNA高通量测序分析大泷六线鱼表皮粘液及肠道内容物微生物多样性

大泷六线鱼(Hexagrammos otakii)是一种具有代表性的高价值冷水性鱼类。通过分析大泷六线鱼表皮粘液及肠道内容物微生物菌群特征,了解其微生物多样性及所携带的自身特有的潜在病原微生物情况。采集设施化车间养殖的大泷六线鱼,提取表皮粘液和肠道内容物基因组DNA,通过16S rRNA高通量测序技术和生物学分析方法,对不同样本的V3+V4区基因文库进行分析。结果表明,大泷六线鱼表皮粘液和肠道内容物样本拥有共同的OTUs为33个,且Venn图呈现了不同来源样本之间的相似性和差异性。Rank-abundance曲线显示了不同样本的均匀度和丰富度,测序数据合理、可信。Alpha指数平均值显示肠道内容物微生物丰富度较高,而表皮粘液微生物多样性较高;Beta多样性显示了不同来源样本间的异质性及同一来源样本间的相似性。从门水平上看,优势菌门均为蓝菌门（Cyanobacteria）、变形菌门（Proteobacteria）、厚壁菌门（Firmicutes）;而属水平结果不同,肠道内容物样本中优势菌属包括Streptophyta、芽孢杆菌属（Bacillus）、杆菌属（Bacillaceae_unclassified）、气单胞菌属（Aeromonas）及弧菌属（Vibrio）,表皮粘液样本中优势菌属包括Streptophyta、气单胞菌属（Aeromonas）、杆菌属（Bacillaceae_unclassified）、香味菌属（Myriudes）、假单胞菌属（Gemmobacter）及弧菌属（Vibrio）。Heatmap热图结果提示,不同微生物菌群结构发挥出不同生物学功能,其中未分类杆菌属、气单胞菌属、弧菌属、假单胞菌属等条件性致病菌的存在均可能导致病害的发生。因此,揭示表皮粘液和肠道内容物样本中微生物的多样性及主要病原菌属的特征对大泷六线鱼健康养殖和疾病防控具有一定的指导意义。     

室温条件下虎斑颈槽蛇卵的孵化与幼蛇饲养

目的探讨室温条件下虎斑颈槽蛇的孵化及幼蛇饲养情况。方法选用同一条虎斑颈槽蛇产的同窝蛇卵31枚,随机为A组（实验组）、B组（对照组）,A组16枚,B组15枚。两组蛇卵分别置于2个完全相同的打孔整理箱中进行孵化,孵出幼蛇用小泽蛙饲养。结果在室温条件下,孵化期58天,孵化率为96．77％。出壳后7～9天,幼蛇第1次蜕皮,蜕皮周期在11～16天不等。结论对虎斑颈槽蛇进行工厂化人工养殖是切实可行的。     

虎斑颈槽蛇治化疗白细胞减少1例

虎斑颈槽蛇治化疗白细胞减少１例吴卯斌安徽省黄山市屯溪毒蛇养殖研究所２４５０５１虎斑颈槽蛇（Ｒｂａｂｄｏｐｈｉｓｔｉｇｙｉｈａｂｏｉｃ）,原名：虎斑游蛇,是有毒蛇,笔者用其治愈化疗期白细胞减少１例。现介绍如下：患者何某,男,４６岁,农民。因自觉食道梗阻...     

鲁西地区蛇类的分布格局

以山东聊城分布的7种蛇类作为研究对象,探讨了鲁西平原区蛇类的群落结构和分布格局.结果 表明,该区蛇类平均分布密度较高的是白条锦蛇Elaphe dione、赤链蛇Dinodon rufozonatum和虎斑颈槽蛇大陆亚种Rhabdophis tigrina lateralis.多数蛇类倾向于选择湿地周边、边缘富有灌草丛的农田区以及有丰富草被的居民区等为栖息地,该区蛇类的栖息地选择有着显著的不重叠性.     

斑头雁成鸟与雏鸟泄殖腔微生物的对比分析

肠道微生物通过维持稳态、辅助消化和促进免疫系统发育等方式维护宿主的健康状态。肠道微生物本身则受到宿主的基因、饮食、年龄和环境等因素的影响。然而,肠道微生物的变化与宿主年龄之间的关系仍有许多未知。本研究分别收集斑头雁(Anser indicus)2只成鸟及3只雏鸟泄殖腔样品,提取肠道微生物总DNA,采用16S rRNA高通量测序的方法,分析并比较两年龄阶段鸟类肠道微生物的菌群结构及组成差异。研究发现,斑头雁雏鸟泄殖腔微生物属于9个门,含量最高的前5个门分别是梭杆菌门(48.29%)、厚壁菌门(22.21%)、变形杆菌门(22.07%)、放线菌门(5.02%)和软壁菌门(1.93%)。成鸟泄殖腔微生物属于17个门,最多的依次是变形菌门(64.69%)、厚壁菌门(23.92%)、蓝细菌(8.48%)、放线菌门(1.43%)和梭杆菌门(0.56%)。在属的水平,斑头雁雏鸟泄殖腔微生物属于18个属,而成鸟含有24个属。成鸟泄殖腔微生物的α多样性显著高于雏鸟(P0.05,Welch′s t-test)。有186个操作分类单元(OTU)属于成鸟和雏鸟共有,而其他640个OTU和90个OTU则分别隶属于成鸟和雏鸟。雏鸟中67.39%的OTUs是成鸟所具有的。基于OTU的聚类结果与年龄分组一致。本结果对认识鸟类肠道微生物与宿主年龄变化之间的关系有一定的参考价值。     

Dysbacteriosis in patients with colon polyposis

The study revealed the most profound changes in the composition of intestinal microflora in patients with polyposis of the large intestine. In these patients anaerobic microflora (bifidobacteria, lactic acid bacteria) was more often suppressed than in other examined groups, in particular, patients with cholelithic disease. The associations of hemolytic Escherichia with Klebsiella pneumoniae, Pseudomonas aeruginosa, Citrobacter freundii were often observed as well as the increased content of enterococci and fungi of the genus Candida. The determination of frequency and degree of manifestations showed that dysbacteriosis was registered in the absolute majority of patients (97.4%) with polyposis of the large intestine. Among patients with cholelithic disease disturbances in microbiocenosis were detected in 60.0% of cases, the profundity and character of the microflora composition changes being less pronounced.     

Microbial-host interactions specifically control the glycosylation pattern in intestinal mouse mucosa

The glycosylation of the intestinal cell layer is thought to control several key functions of the gut such as vectorial transports, defence against microbial agents or immunological processes. It has been assumed that the gut microflora may modulate the glycosylation pattern of the intestinal cell layer. However, there is no direct evidence for this regulatory process. The first goal of this work was to establish the germ-free mice intestinal glycosylation baseline using a histochemical approach and a panel of ten lectins with defined glycan specificities to tissue sections prepared from various cellular compartments of the small and large intestine. Using this baseline, we have studied the contribution of the gut microflora on the carbohydrate composition of glycoconjugates of intestinal cells by comparing the germ-free and conventional mice glycosylation patterns. Analysis of the germ-free mice intestinal glycosylation baseline revealed that the expression of glycans depends on the proximodistal gradient (small to large intestine) and on the cell lineage (absorptive, goblet, crypt, and Paneth cells), indicating that mice are able to create and maintain a strict topological and cell lineage-specific regulation of glycosyltransferase expression. By comparing germ-free and conventional mice, we find that the gut microflora specifically modulates the gut glycosylation pattern, quantitatively as well as qualitatively by changing the cellular and subcellular distribution of glycans. This is the first report in mice to directly demonstrate the critical contribution of microflora to intestinal glycosylation, a key characteristic of the gut.     

基于高通量测序的褐飞虱肠道微生物多样性分析

【目的】探明褐飞虱Nilaparvata lugens成虫肠道微生物群落结构和多样性。【方法】分离褐飞虱成虫完整肠道并提取总DNA,利用Illumina MiSeq(PE300)技术对其肠道细菌16S rRNA的V3-V4变异区和真菌ITS2序列进行测序,统计肠道微生物的操作分类单元(operational taxonomic unit, OTU)数量,分析其物种组成、丰度及Alpha多样性。并通过qPCR技术验证随机挑选注释到的4种肠道菌的高通量测序数据的有效性。【结果】分别获得褐飞虱成虫肠道细菌16S rRNA和真菌ITS2优质序列32 395和24 986条,根据序列相似性进行聚类分析分别获得235和128个OTUs。其中,细菌共注释到7个门, 15个纲, 26个目, 45个科和73个属;真菌共鉴定到3个门, 9个纲, 12个目, 15个科和18个属。在门分类水平上,细菌以变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)为优势门类;真菌以子囊菌门(Ascomycota)为绝对优势菌门。在属分类水平上,细菌的优势属为不动杆菌属Acinetobacter以及紫单胞菌科(Porphyromonadaceae)未确定属和毛螺菌科(Lachnospiraceae)未确定属,其丰度分别为36.37%, 17.22%和15.01%;真菌的优势属为粪壳菌纲(Sordariomycetes)未确定属,丰度为95.77%。Alpha多样性分析结果显示,褐飞虱肠道细菌(真菌)的观测物种数、Chao1指数、Shannon指数和Simpson 指数分别为235(128), 262.64(165.40), 3.90(0.91)和0.62(0.75)。4种肠道菌的qPCR结果显示高通量测序数据具有较高的有效性。【结论】褐飞虱成虫肠道细菌和真菌群落整体多样性比较丰富。研究结果为从共生微生物角度解析褐飞虱的环境适应性以及开发基于微生物防治的新技术等方面提供了依据。     

Anoxic Conditions Promote Species-Specific Mutualism between Gut Microbes In Silico

The human gut is inhabited by thousands of microbial species, most of which are still uncharacterized. Gut microbes have adapted to each other''s presence as well as to the host and engage in complex cross feeding. Constraint-based modeling has been successfully applied to predicting microbe-microbe interactions, such as commensalism, mutualism, and competition. Here, we apply a constraint-based approach to model pairwise interactions between 11 representative gut microbes. Microbe-microbe interactions were computationally modeled in conjunction with human small intestinal enterocytes, and the microbe pairs were subjected to three diets with various levels of carbohydrate, fat, and protein in normoxic or anoxic environments. Each microbe engaged in species-specific commensal, parasitic, mutualistic, or competitive interactions. For instance, Streptococcus thermophilus efficiently outcompeted microbes with which it was paired, in agreement with the domination of streptococci in the small intestinal microbiota. Under anoxic conditions, the probiotic organism Lactobacillus plantarum displayed mutualistic behavior toward six other species, which, surprisingly, were almost entirely abolished under normoxic conditions. This finding suggests that the anoxic conditions in the large intestine drive mutualistic cross feeding, leading to the evolvement of an ecosystem more complex than that of the small intestinal microbiota. Moreover, we predict that the presence of the small intestinal enterocyte induces competition over host-derived nutrients. The presented framework can readily be expanded to a larger gut microbial community. This modeling approach will be of great value for subsequent studies aiming to predict conditions favoring desirable microbes or suppressing pathogens.     

The Dynamic Distribution of Porcine Microbiota across Different Ages and Gastrointestinal Tract Segments

Metagenome of gut microbes has been implicated in metabolism, immunity, and health maintenance of its host. However, in most of previous studies, the microbiota was sampled from feces instead of gastrointestinal (GI) tract. In this study, we compared the microbial populations from feces at four different developmental stages and contents of four intestinal segments at maturity to examine the dynamic shift of microbiota in pigs and investigated whether adult porcine fecal samples could be used to represent samples of the GI tract. Analysis results revealed that the ratio of Firmicutes to Bacteroidetes from the feces of the older pigs (2-, 3-, 6- month) were 10 times higher compared to those from piglets (1-month). As the pigs matured, so did it seem that the composition of microbiome became more stable in feces. In adult pigs, there were significant differences in microbial profiles between the contents of the small intestine and large intestine. The dominant genera in the small intestine belonged to aerobe or facultative anaerobe categories, whereas the main genera in the large intestine were all anaerobes. Compared to the GI tract, the composition of microbiome was quite different in feces. The microbial profile in large intestine was more similar to feces than those in the small intestine, with the similarity of 0.75 and 0.38 on average, respectively. Microbial functions, predicted by metagenome profiles, showed the enrichment associated with metabolism pathway and metabolic disease in large intestine and feces while higher abundance of infectious disease, immune function disease, and cancer in small intestine. Fecal microbes also showed enriched function in metabolic pathways compared to microbes from pooled gut contents. Our study extended the understanding of dynamic shift of gut microbes during pig growth and also characterized the profiles of bacterial communities across GI tracts of mature pigs.     

Bacteria,phages and pigs: the effects of in-feed antibiotics on the microbiome at different gut locations

Disturbance of the beneficial gut microbial community is a potential collateral effect of antibiotics, which have many uses in animal agriculture (disease treatment or prevention and feed efficiency improvement). Understanding antibiotic effects on bacterial communities at different intestinal locations is essential to realize the full benefits and consequences of in-feed antibiotics. In this study, we defined the lumenal and mucosal bacterial communities from the small intestine (ileum) and large intestine (cecum and colon) plus feces, and characterized the effects of in-feed antibiotics (chlortetracycline, sulfamethazine and penicillin (ASP250)) on these communities. 16S rRNA gene sequence and metagenomic analyses of bacterial membership and functions revealed dramatic differences between small and large intestinal locations, including enrichment of Firmicutes and phage-encoding genes in the ileum. The large intestinal microbiota encoded numerous genes to degrade plant cell wall components, and these genes were lacking in the ileum. The mucosa-associated ileal microbiota harbored greater bacterial diversity than the lumen but similar membership to the mucosa of the large intestine, suggesting that most gut microbes can associate with the mucosa and might serve as an inoculum for the lumen. The collateral effects on the microbiota of antibiotic-fed animals caused divergence from that of control animals, with notable changes being increases in Escherichia coli populations in the ileum, Lachnobacterium spp. in all gut locations, and resistance genes to antibiotics not administered. Characterizing the differential metabolic capacities and response to perturbation at distinct intestinal locations will inform strategies to improve gut health and food safety.     

基于16S rRNA基因高通量测序的 太原地区健康成年人肠道菌群结构分析

目的探讨太原地区健康成年人的肠道菌群结构及多样性。方法采集太原地区20份健康成年人的粪便样本,提取DNA、构建菌群16S rRNA基因克隆文库、高通量测序并进行生物信息学分析。结果测序获得优质序列1 383 680条,平均序列为69 184条±551条,归类于455个OTUs;所有样本共含有10个菌门,101个菌属,141种菌;其中核心菌门3个,依次为拟杆菌门(63.13%)、厚壁菌门(31.14%)和变形菌门(5.10%),占所有样本微生物总量的99.37%;丰度最高的前10个菌属依次为拟杆菌属(43.34%)、普氏菌属(15.51%)、栖粪杆菌属(4.92%)、罗氏菌属(4.73%)、毛螺菌属(3.27%)、萨特菌属(2.50%)、粪球菌属(2.38%)、布劳特菌属(2.31%)、瘤胃球菌属(2.18%)和副杆状菌属(1.61%),占样本微生物总量的82.75%,未分类的菌属占6.84%。结论健康成年人肠道微生物群落复杂,但主要菌群相对稳定,为进一步研究人体肠道菌群结构与功能提供了参考依据。