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Recovery of different Listeria ribotypes from naturally contaminated raw refrigerated meat and poultry products with two primary enrichment media.

机译:使用两种主要的富集培养基从天然污染的原始冷藏肉类和家禽产品中回收不同的李斯特菌核糖型。

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摘要

Isolation rates for Listeria monocytogenes and the other Listeria spp. typically improve when samples are enriched in more than one primary enrichment medium. This study evaluated the abilities of two primary enrichment media, University of Vermont-modified Listeria enrichment broth (UVM) and Listeria repair broth (LRB), to recover different ribotypes of Listeria spp. from raw meat and poultry samples. Forty-five paired 25-g retail samples of ground beef, pork sausage, ground turkey, and chicken (160 samples) underwent primary enrichment in UVM and LRB (30 degrees C for 24 h) followed by secondary enrichment in Fraser broth (35 degrees C for 24 and 40 h) and plating on modified Oxford agar. After 24 h of incubation of 35 degrees C, 608 Listeria colonies from selected positive samples were biochemically confirmed as L. monocytogenes (245 isolates), L innocua (276 isolates), and L. welshimeri (89 isolates) and then ribotyped with the automated Riboprinter microbial characterization system (E. I. du Pont de Nemours & Co., Inc.). Thirty-six different Listeria strains comprising 16 L. monocytogenes (including four known clinical ribotypes), 12 L. innocua, and 8 L. welshimeri ribotypes were identified from selected positive samples (15 samples of each product type; two UVM and two LRB isolates per sample). Twenty-six of 36(13 L. monocytogenes) ribotypes were detected with both UVM and LRB, whereas 3 of 36 (1 L. monocytogenes) and 7 of 36 (3 L. monocytogenes) Listeria ribotypes were observed with only UVM or LRB, respectively. Ground beef, pork sausage, ground turkey, and chicken yielded 22 (8 L. monocytogenes), 21 (12 L. monocytogenes), 20 (9 L. monocytogenes), and 19 (11 L. monocytogenes) different Listeria ribotypes, respectively, with some Listeria ribotypes confined to a particular product. More importantly, major differences in both the number and distribution of Listeria ribotypes, including previously recognized clinical and nonclinical ribotypes of L. monocytogenes, were observed when 10 UVM and 10 LRB isolates from five samples of each product were ribotyped. When a third set of six samples per product type was examined from which two Listeria isolates were obtained by using only one of the two primary enrichment media, UVM and LRB failed to detect L. monocytogenes (both clinical and nonclinical ribotypes) in two and four samples, respectively. These findings stress the importance of using more than one primary enrichment medium and picking a sufficient number of colonies per sample when attempting to isolate specific L. monocytogenes strains during investigations of food-borne listeriosis.
机译:李斯特菌和其他李斯特菌属的分离率。当样品在一种以上的主要富集培养基中富集时,通常会改善。这项研究评估了两种主要的富集培养基,即佛蒙特大学改良的李斯特菌富集肉汤(UVM)和李斯特菌修复肉汤(LRB),可以恢复李斯特菌属的不同核糖型。从生肉和家禽样品中提取。 25对成对的25克零售牛肉,猪肉香肠,火鸡和鸡肉的25 g样品(160个样品)进行了UVM和LRB的初步富集(30摄氏度,持续24小时),然后在Fraser肉汤中进行了二次富集(35摄氏度) C为24和40小时),并在改性牛津琼脂上铺板。在35°C下孵育24小时后,通过生化方法从选定的阳性样本中筛选出608个李斯特菌菌落,分别为单核细胞增生李斯特菌(245个分离株),无毒李斯特菌(276个分离株)和韦氏乳杆菌(89个分离株),然后用自动核糖酶进行分型Riboprinter微生物鉴定系统(EI du Pont de Nemours&Co.,Inc.)。从选定的阳性样品中鉴定了36种不同的李斯特菌菌株,包括16种单核细胞增生李斯特菌(包括4种已知的临床核糖型),12种无毒李斯特菌和8个韦氏乳杆菌核糖型(每种产品类型15个样品;两个UVM和两个LRB分离株)每个样本)。仅使用UVM或LRB时,使用UVM和LRB检测到36种(13个单核细胞增生李斯特菌)核糖型中的26种,而仅使用UVM或LRB的李斯特菌核糖型分别观察到36种(1个单核细胞增生李斯特菌)的3种和36种(3个单核细胞增生李斯特菌)的7种。分别。碎牛肉,猪肉香肠,火鸡和鸡肉分别产生22种(单核细胞增生李斯特菌8种),21种(单核细胞增生李斯特菌12种),20种(单核细胞增生李斯特菌9种)和19种(单核增生李斯特菌引起的)19种核糖核酸。某些李斯特菌核糖型仅限于特定产品。更重要的是,当对每种产品的五个样品中的10个UVM和10个LRB分离株进行核糖分型时,观察到李斯特菌核糖型数和分布的主要差异,包括以前公认的单核细胞增生李斯特菌的临床和非临床核糖型。当检查每种产品类型的第三组六个样品时,仅使用两种主要富集培养基中的一种从中获得了两个李斯特菌分离株,UVM和LRB无法在两个和四个中检测到单核细胞增生李斯特菌(临床和非临床核型)。样本。这些发现强调了在食源性李斯特菌病调查过程中尝试分离特定的单核细胞增生李斯特菌菌株时,必须使用一种以上的富集培养基并在每个样品中选择足够数量的菌落的重要性。

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