Laciar A L, Picca S, de Centorbi O P
Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis, Argentina.
Rev Argent Microbiol. 1994 Oct-Dec;26(4):183-8.
Recent food listeriosis outbreaks confirm that more faithful isolation and identification methods for Listeria monocytogenes or other potentially pathogen microorganisms are required. Furthermore, the human and animal reservoir role in the ecology of this disease must be established. Listeria spp. in the vizcacha intestinal content was determined by two isolation procedures, starting from 10 g of homogenized samples in 40 ml of PBS. I)0.1 ml was stripped on phenylethanol agar, selective agar for Listeria and acryflavin ceftazidin agar, then incubated at 37 degrees C for 48 h, suspected colonies were identified by preliminary tests (Gram, hemolysis, catalase, esculin hydrolisis and motility at 22 degrees C) and confirmatory tests (indol, methyl red, Voges Proskauer, nitrate and carbohydrate fermentation) (Table 1). Antibiotic susceptibility, protein profile by PAGE and pathogenic power in mice were determined. II) The remaining homogenate was incubated at 4 degrees C in 100 ml of Donnelly and Baigent enrichment broth, weekly or monthly with subcultures until 30 days or 6-8 months, respectively. The subcultures were followed up as in I). A L. seeligeri strain, susceptible to antibiotics suggested for L. monocytogenes and exhibiting resistance to some second and third generation cephalosporins, was isolated (Table 2). The protein profile of both species was coincident, but L. seeligeri was not virulent for mice. The finding of L. seeligeri in an animal (4.0%) used as human feeding source is of interest due to its potential pathogen power.
近期的食源性李斯特菌病暴发表明,需要更可靠的单核细胞增生李斯特菌或其他潜在致病微生物的分离和鉴定方法。此外,必须明确人和动物宿主在这种疾病生态中的作用。从40毫升磷酸盐缓冲液中10克匀浆样品开始,通过两种分离程序测定了草原鼠兔肠道内容物中的李斯特菌属。I)将0.1毫升接种在苯乙醇琼脂、李斯特菌选择性琼脂和吖啶黄素头孢他啶琼脂上,然后在37℃下孵育48小时,通过初步试验(革兰氏染色、溶血、过氧化氢酶、七叶苷水解和22℃下的运动性)和确证试验(吲哚、甲基红、Voges Proskauer试验、硝酸盐和碳水化合物发酵)鉴定疑似菌落(表1)。测定了抗生素敏感性、聚丙烯酰胺凝胶电泳蛋白质谱和对小鼠的致病力。II)将剩余的匀浆在4℃下于100毫升唐纳利和贝金特增菌肉汤中孵育,分别每周或每月传代培养,直至30天或6 - 8个月。传代培养按照I)中的方法进行后续观察。分离出了一株对推荐用于单核细胞增生李斯特菌的抗生素敏感,但对一些第二代和第三代头孢菌素耐药的西利格李斯特菌(表2)。两种菌的蛋白质谱一致,但西利格李斯特菌对小鼠无致病性。鉴于其潜在致病力,在作为人类食物来源的动物(4.0%)中发现西利格李斯特菌令人关注。