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Isolation, characterization and comparison of bacteria from swine faeces and manure storage pits

机译:猪粪和粪便粪便中细菌的分离,鉴定和比较

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摘要

Storage of swine manure is associated with the microbiological production of a variety of odorous chemicals including ammonia, organic acids and alcohols, and sulphides. Although largely the product of microbiological activity, little is known about the microorganisms present in swine manure. In order to gain a better understanding of the types and activities of the microorganisms present, representative strains of microorganisms were isolated from faeces and stored manure slurry, identified, and physiologically characterized. For swine manure slurry samples, total anaerobe colony counts were greatest when a non-selective, habitat simulating medium containing clarified swine manure slurry was used whereas the highest counts for faecal anaerobes were obtained on rumen fluid containing medium. Faecal and slurry samples were also plated onto the appropriate medium containing the antibiotics tetracycline, erythromycin and tylosin (10 μg ml~(-1), individually) and the proportional counts of organisms capable of growing in the presence of these antibiotics determined. Randomly selected isolates from the highest dilutions were identified physiological characteristics were determined. The results of these examinations indicate that the predominant culturable microorganisms from these environments are oblately anaerobic, low mol percentage G + C Gram positive bacteria (Firmicutes) who are members of Clostridial, Eubacterial, and Lactobacillus/Streptococcus phylogenetic groups. Isolates similar to Sporomusa and Flexibacter/Cytophaga/Bacteroides (CFB or Bacteroidetes) groups were also obtained. Although similar overall, faecal and slurry samples differed in bacterial composition. Manure slurry samples were dominated by organisms similar to Clostridium coccoides and Enterococcus species whereas the distribution of species present in faeces appeared much broader. Whereas most of the pure cultures could be assigned to known phylogenetic groupings, few could be identified as known species. Examination of some growth and physiological characteristics of faecal and slurry isolates showed these to be primarily carbohydrate fermenters, although some were able to ferment lactate and amino acids. When the ability of manure and faecal isolates to ferment protein, peptides and amino acids was examined, a relatively small percentage of these were able to do so and most of these fermented carbohydrates is addition to the amino acid sources provided. The predominant amino acid fermenters were most closely related to C. coccoides and C. botulinum, but representatives of the Bacteroides, Staphylococcus, Enterococcus and other phylogenetic groups were also found. The results reported here are compared with those obtained from clone libraries prepared from the same environmental samples.
机译:猪粪的储存与各种有气味的化学物质的微生物产生有关​​,这些化学物质包括氨,有机酸和醇以及硫化物。尽管很大程度上是微生物活性的产物,但对猪粪中存在的微生物知之甚少。为了更好地了解存在的微生物的类型和活性,从粪便中分离出代表性的微生物菌株,并贮存粪便,对其进行鉴定和生理学表征。对于猪粪浆样品,当使用含有澄清猪粪浆的非选择性生境模拟培养基时,总厌氧菌落计数最高,而在含有瘤胃液的培养基中粪便厌氧菌计数最高。粪便和浆液样品也被接种到含有抗生素四环素,红霉素和泰乐菌素(分别为10μgml〜(-1))的合适培养基上,并确定在这些抗生素存在下能够生长的生物体的比例计数。确定了从最高稀释度中随机选择的分离物,确定了生理特征。这些检查的结果表明,来自这些环境的主要可培养微生物是厌氧,低摩尔百分率的G + C革兰氏阳性细菌(Firmicutes),它们是梭菌,真细菌和乳杆菌/链球菌系统发生群的成员。还获得了类似于孢子虫和弯曲杆菌/噬菌丝/拟杆菌(CFB或拟杆菌)组的分离株。尽管总体上相似,但粪便和浆液样品的细菌组成有所不同。粪便粪便样品被类似于球孢梭菌和肠球菌物种的生物所控制,而粪便中存在的物种分布似乎更为广泛。尽管大多数纯培养物可以分配到已知的系统发生组中,但很少可以鉴定为已知的种。对粪便和浆液分离物的某些生长和生理特性的检查表明,它们主要是碳水化合物发酵罐,尽管有些能够发酵乳酸和氨基酸。当检查粪便和粪便分离物发酵蛋白质,肽和氨基酸的能力时,其中相对较小的比例能够做到这一点,并且这些发酵的碳水化合物中的大多数是所提供的氨基酸来源的补充。主要的氨基酸发酵罐与C.coccoides和C.肉毒杆菌最密切相关,但同时也发现了拟杆菌,葡萄球菌,肠球菌和其他系统发生群。将此处报告的结果与从相同环境样品制备的克隆文库中获得的结果进行比较。

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