产气荚膜梭菌α、β毒素研究进展

产气荚膜梭菌(Clostridium perfringens)是一种革兰氏阳性、产生芽胞、严格厌氧及形成特殊荚膜的梭状杆菌。该菌广泛存在于自然界的水源、土壤及人和动物肠道之中。研究发现,该菌至少可以产生15种以上的毒素,但目前仅有5-6种毒素,即α、β、ε、ι毒素及肠毒素(CPE)、β2毒素等被认为是该菌的主要致病性毒素。根据产气荚膜梭菌产生四种致死性毒素(α、β、ε、ι毒素)的能力,将其分为五种类型,即A、B、C、D、E型。产气荚膜梭菌是一种重要的人畜共患病的病原体,可引起人的食物中毒和多种动物坏死性肠炎、肠毒血症同时也是人畜创伤性气性坏疽以及人类食物中毒的主要病原菌之一。近年来,对产气荚膜梭菌α、β毒素研究取得了重大进展。现就产气荚膜梭菌α、β毒素的组成、结构、分子生物学、检测等方面的研究进展做一综述。     

Comparison of conventional and molecular techniques to investigate the intestinal microflora of rainbow trout (Oncorhynchus mykiss)

The concept of the use of probiotic organisms or prebiotic compounds to modify the fish gut microflora is becoming a popular topic for investigation. A major flaw in many such studies is a failure to consider fully the nature of the established microflora, which is to be modified pre-, or probiotically. Since it is widely accepted that a large proportion of bacteria are non-culturable, the use of conventional bacteriological (culture) techniques alone to investigate fish intestinal microflora may be expected to bias results. We report a study designed to investigate the normal intestinal microflora of rainbow trout (Oncorhynchus mykiss) using both conventional bacteriological and molecular methods.Over an 18 month period, the intestinal microflora of a single population of laboratory-raised rainbow trout was investigated. Bacteria isolated using bacteriological techniques were identified using the BiOLOG system and 16S rRNA gene sequencing. Dominant bacteria consistently were Aeromonas sp. and Carnobacterium piscicola, demonstrating that the microflora is stable in fish kept in defined conditions. Restriction Fragment Length Polymorphism (RFLP) and 16S rRNA gene sequence analysis was used to investigate anaerobic and non-culturable bacteria. An obligate anaerobe, Clostridium gasigenes, was shown to be among the dominating intestinal microflora.     

产气荚膜梭菌多重PCR定型方法的建立及其应用

建立了多重PCR检测产气荚膜梭菌α、β、ε和ι毒素基因的方法。该方法具有良好的特异性和敏感性,只有产气荚膜梭菌呈现阳性,被检验的其他梭菌以及大肠杆菌、葡萄球菌均为阴性;将肉汤菌液样品10倍系列稀释后进行检测,检测灵敏度达到1.2×104CFU/mL。收集40份牛粪便样品,进行PCR检测,32份样品中成功扩增出589 bp的α毒素基因片段,阳性率为80%。结果显示,建立的多重PCR检测方法可取代血清中和试验,用于产气荚膜梭菌分型,同时表明A型产气荚膜梭菌在当地奶牛场中较为普遍。     

Clostridium difficile in faeces from healthy dogs and dogs with diarrhea

Background

This study was conducted to evaluate the faecal occurrence and characterization of Clostridium difficile in clinically healthy dogs (N = 50) and in dogs with diarrhea (N = 20) in the Stockholm-Uppsala region of Sweden.

Findings

Clostridium difficile was isolated from 2/50 healthy dogs and from 2/20 diarrheic dogs. Isolates from healthy dogs were negative for toxin A and B and for the tcdA and tcdB genes. Both isolates from diarrheic dogs were positive for toxin B and for the tcdA and tcdB genes. The C. difficile isolates from healthy dogs had PCR ribotype 009 (SE-type 6) and 010 (SE-type 3) whereas both isolates from dogs with diarrhoea had the toxigenic ribotype 014 (SE-type 21). One of the isolates from healthy dogs was initially resistant to metronidazole.

Conclusions

This study revealed presence of toxigenic C. difficile in faecal samples of diarrheic dogs and low number of non- toxigenic isolates in healthy dogs from Uppsala-Stockholm region in Sweden. However, more comprehensive studies are warranted to investigate the role of C. difficile in gastrointestinal disease in dogs.     

Effect of Vaccination Against Coccidiosis in Combination with an Antibacterial Oregano (Origanum vulgare) Compound in Organic Broiler Production

Effects on production and health in broilers vaccinated against coccidiosis (Paracox®, Schering-Plough Animal Health) in combination with an antibacterial organic compound (Orego-Stim®, Meriden Animal Health Ltd.) were investigated. To obtain an environment and infection level similar to practical conditions, experimental floor pens were set up in a commercial broiler chicken house. Orego-Stim® supplement increased live weight and feed intake up to 48 days of age, whilst feed conversion ratio was not significantly affected. Vaccinated birds had a significantly higher live weight between 34 and 48 days, compared to non-vaccinated, but vaccination did not significantly affect feed intake or feed conversion ratio. Coccidial numbers in the litter were low, and only minor intestinal lesions were found. There were no significant differences in lesion scores between the different treatments. Orego-Stim® supplement decreased caecal numbers of Clostridium perfringens at 31 days, but not at 52 days, and numbers were not affected at all by anticoccidial vaccination. Bird health was overall good. The results indicate that vaccination against coccidiosis in combination with an in-feed antibacterial compound may be an alternative control method for intestinal health in organically produced chickens.     

鹑梭状芽胞杆菌外毒素的分离与特性研究

用色氨酸磷酸液体培养基厌氧培养， 蔡氏滤器过滤， 以饱和硫酸铵沉淀， 制备了鹑梭状芽胞杆菌外毒素， 经检测该毒素的稳定性较差， ６０ ℃１０ 分钟即可破坏， 经 Ｓ Ｄ Ｓ聚丙烯酰胺凝胶电泳分析证明， 该毒素至少有１１种蛋白成分， 分子量在３００００ ～６７０００ 之间， 静脉注射小鼠０２ ｍ ｇ 可引起死亡， 经口灌服雏鸡， 可出现肝脏坏死和肠粘膜溃疡。     

牛猝死症诊断和防制

经流行病学调查,病原分离鉴定,病理学检查,临床症状观察确定黑龙江省猝死症的主要病原为Ａ,Ｃ型魏氏梭菌。在部分死亡病例中,也有因误食灭鼠药氟乙酰胺而引起的中毒,但数量较少,根据病原分离结果,魏氏梭菌和腐败梭菌,魏氏梭菌和巴氏杆菌,魏氏杆菌和链球菌可形成混合感染,使魏氏梭菌病疫情更加严重而呈地方性流行,这是近几年来牛魏氏梭菌病的新特点。     

牛魏氏梭菌病概述

魏氏梭菌(Clostridium welchii),又称产气荚膜梭菌(Clostridium perfringens),是引起牛的猝死病的主要病原,对养牛业的危害极大。本文简要介绍了牛魏氏梭菌病的病原、流行病学、临床症状、病理变化、诊断和防制。     

魏氏梭菌α毒素基因的克隆表达及其间接ELISA方法的建立

对GenBank上发表的魏氏梭菌α毒素基因的氨基酸序列进行了二级结构、疏水性、抗原决定簇、跨膜区以及抗原性分析,在此基础上,对α毒素基因130～966nt位的837bp长的适合原核表达的区域进行PCR扩增、克隆、核酸序列测定和生物信息学分析。同时,建立了检测魏氏梭菌α毒素抗体的间接ELISA方法。