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培养温度对从普雷斯顿肉汤增菌的食品中分离空肠弯曲菌基因型的影响。

Effect of incubation temperature on isolation of Campylobacter jejuni genotypes from foodstuffs enriched in Preston broth.

作者信息

Scates Pam, Moran Lynn, Madden Robert H

机构信息

Food Microbiology Branch,Department of Agriculture and Rural Development. Food Science Department, Queen's University of Belfast, Belfast BT9 5PX, Northern Ireland.

出版信息

Appl Environ Microbiol. 2003 Aug;69(8):4658-61. doi: 10.1128/AEM.69.8.4658-4661.2003.

Abstract

Preston broth and agar incubated at either 37 or 42 degrees C have been widely used to isolate campylobacters from foodstuffs. The consequences of using either incubation temperature were investigated. Retail packs of raw chicken (n = 24) and raw lamb liver (n = 30) were purchased. Samples were incubated in Preston broth at 37 and 42 degrees C and then streaked onto Preston agar and incubated as before. Two Campylobacter isolates per treatment were characterized. Poultry isolates were genotyped by random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE), and flagellin PCR-restriction fragment length polymorphism, and lamb isolates were genotyped by RAPD only. In total, 96% of the poultry and 73% of the lamb samples yielded campylobacters. The lamb isolates were all Campylobacter jejuni, as were 96% of the poultry isolates, with the remainder being Campylobacter lari. The incubation temperature had no significant effect on the number of positive samples or on the species isolated. However, genotyping of the C. jejuni isolates revealed profound differences in the types obtained. Overall (from poultry and lamb), the use of a single incubation temperature, 37 degrees C, gave 56% of the total number of RAPD C. jejuni genotypes, and hence, 44% remained undetected. The effect was especially marked in the poultry samples, where incubation at 37 degrees C gave 47% of the PFGE genotypes but 53% were exclusively recovered after incubation at 42 degrees C. Thus, the incubation temperature of Preston media selects for certain genotypes of C. jejuni, and to detect the widest range, samples should be incubated at both 37 and 42 degrees C. Conversely, genotyping results arising from the use of a single incubation temperature should be interpreted with caution.

摘要

将普雷斯顿肉汤和琼脂在37或42摄氏度下孵育,已被广泛用于从食品中分离弯曲杆菌。研究了使用这两种孵育温度的后果。购买了零售包装的生鸡肉(n = 24)和生羊肝(n = 30)。将样品在普雷斯顿肉汤中于37和42摄氏度下孵育,然后划线接种到普雷斯顿琼脂上并按之前的方式孵育。对每种处理的两个弯曲杆菌分离株进行了鉴定。家禽分离株通过随机扩增多态性DNA(RAPD)、脉冲场凝胶电泳(PFGE)和鞭毛蛋白PCR-限制性片段长度多态性进行基因分型,而羊分离株仅通过RAPD进行基因分型。总体而言,96%的家禽样品和73%的羊样品产生了弯曲杆菌。羊分离株均为空肠弯曲杆菌,家禽分离株中96%也是空肠弯曲杆菌,其余为拉氏弯曲杆菌。孵育温度对阳性样品数量或分离出的菌种没有显著影响。然而,空肠弯曲杆菌分离株的基因分型显示所获得的类型存在显著差异。总体而言(来自家禽和羊),使用单一孵育温度37摄氏度,得到了RAPD空肠弯曲杆菌基因型总数的56%,因此,44%仍未被检测到。这种影响在家禽样品中尤为明显,在37摄氏度下孵育得到了47%的PFGE基因型,但在42摄氏度下孵育后才完全获得了53%的基因型。因此,普雷斯顿培养基的孵育温度选择了某些空肠弯曲杆菌基因型,为了检测到最广泛的范围,样品应在37和42摄氏度下都进行孵育。相反,由单一孵育温度产生的基因分型结果应谨慎解释。

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