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成年猪粪便细菌的分离与鉴定

Isolation and identification of fecal bacteria from adult swine.

作者信息

Salanitro J P, Blake I G, Muirhead P A

出版信息

Appl Environ Microbiol. 1977 Jan;33(1):79-84. doi: 10.1128/aem.33.1.79-84.1977.

Abstract

An examination of the fecal microflora of adult swine was made with regard to the efficiency of several roll tube media in enumeration and recovery of anaerobes, the effects of medium constituents on recovery, and the isolation and identification of the predominant kinds of bacteria. Total number of organisms by microscopic bacterial counts varied among fecal samples from 4.48 X 10(10) to 7.40 X 10(10) bacteria/g (wet weight). Comparison of different nonselective roll tube media indicated that about 30% of the fecal bacteria could be recovered with a rumen fluid (40%, vol/vol) medium (M98-5). Recoveries of 21 and 15%, respectively, were obtained with M10 and rumen fluid-glucose-cellobiose agar (RGCA) media. Rumen fluid, Trypticase, sugars, and CO2 gas phase were important components required for maximum recovery with this medium. Similar high recoveries of anaerobes were also obtained with M98-5 containing swine cecal extract of place in rumen fluid or M10 plus swine cecal extract. Significantly lower recoveries were observed with RCGA, media supplemented with swine fecal extracts, reinforced clostridial medium, brain heart infusion agar, and prereduced blood agar. Ninety percent of the bacteria isolated from roll tube media were gram positive and consisted of facultatively anaerobic streptococci, Eubacterium sp., Clostridium sp., and Propionibacterium acnes. The remainder of the flora (8%) included several other species of anaerobes and Escherichia coli. Rumen fluid (or volatile fatty acids), Trypticase, and yeast extract additions to basal media stimulated the growth of anaerobic strains. Variation in the relative proportions of the predominant fecal microflora was observed. This work indicates that satisfactory enumeration, isolation and cultivation of the predominant microflora in swine feces can be obtained when strict anaerobic culture methods and a rumen fluid medium are used.

摘要

对成年猪粪便微生物区系进行了研究,内容涉及几种滚管培养基在厌氧菌计数和回收方面的效率、培养基成分对回收的影响以及主要细菌种类的分离和鉴定。通过显微镜细菌计数得出的粪便样本中微生物总数在4.48×10¹⁰至7.40×10¹⁰个细菌/克(湿重)之间变化。不同非选择性滚管培养基的比较表明,使用瘤胃液(40%,体积/体积)培养基(M98 - 5)可回收约30%的粪便细菌。使用M10和瘤胃液 - 葡萄糖 - 纤维二糖琼脂(RGCA)培养基分别回收了21%和15%的细菌。瘤胃液、胰蛋白酶、糖类和二氧化碳气相是该培养基实现最大回收所需的重要成分。在瘤胃液中加入猪盲肠提取物的M98 - 5或M10加猪盲肠提取物也获得了类似的高厌氧菌回收率。观察到RCGA、添加猪粪便提取物的培养基、强化梭菌培养基、脑心浸液琼脂和预还原血琼脂的回收率显著较低。从滚管培养基中分离出的细菌90%为革兰氏阳性菌,包括兼性厌氧链球菌、真杆菌属、梭菌属和痤疮丙酸杆菌。其余菌群(8%)包括其他几种厌氧菌和大肠杆菌。向基础培养基中添加瘤胃液(或挥发性脂肪酸)、胰蛋白酶和酵母提取物可刺激厌氧菌株的生长。观察到主要粪便微生物区系相对比例的变化。这项工作表明,当使用严格的厌氧培养方法和瘤胃液培养基时,可以对猪粪便中的主要微生物区系进行令人满意的计数、分离和培养。

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