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rpoS在大肠杆菌O157:H7耐酸性及粪便排出中的作用

Role of rpoS in acid resistance and fecal shedding of Escherichia coli O157:H7.

作者信息

Price S B, Cheng C M, Kaspar C W, Wright J C, DeGraves F J, Penfound T A, Castanie-Cornet M P, Foster J W

机构信息

Departments of Pathobiology, College of Veterinary Medicine, Auburn University, Auburn, Alabama 36849, USA.

出版信息

Appl Environ Microbiol. 2000 Feb;66(2):632-7. doi: 10.1128/AEM.66.2.632-637.2000.

Abstract

Acid resistance (AR) is important to survival of Escherichia coli O157:H7 in acidic foods and may play a role during passage through the bovine host. In this study, we examined the role in AR of the rpoS-encoded global stress response regulator sigma(S) and its effect on shedding of E. coli O157:H7 in mice and calves. When assayed for each of the three AR systems identified in E. coli, an rpoS mutant (rpoS::pRR10) of E. coli O157:H7 lacked the glucose-repressed system and possessed reduced levels of both the arginine- and glutamate-dependent AR systems. After administration of the rpoS mutant and the wild-type strain (ATCC 43895) to ICR mice at doses ranging from 10(1) to 10(4) CFU, we found the wild-type strain in feces of mice given lower doses (10(2) versus 10(3) CFU) and at a greater frequency (80% versus 13%) than the mutant strain. The reduction in passage of the rpoS mutant was due to decreased AR, as administration of the mutant in 0.05 M phosphate buffer facilitated passage and increased the frequency of recovery in feces from 27 to 67% at a dose of 10(4) CFU. Enumeration of E. coli O157:H7 in feces from calves inoculated with an equal mixture of the wild-type strain and the rpoS mutant demonstrated shedding of the mutant to be 10- to 100-fold lower than wild-type numbers. This difference in shedding between the wild-type strain and the rpoS mutant was statistically significant (P </= 0.05). Thus, sigma(S) appears to play a role in E. coli O157:H7 passage in mice and shedding from calves, possibly by inducing expression of the glucose-repressed RpoS-dependent AR determinant and thus increasing resistance to gastrointestinal stress. These findings may provide clues for future efforts aimed at reducing or eliminating this pathogen from cattle herds.

摘要

耐酸性(AR)对于大肠杆菌O157:H7在酸性食物中的存活至关重要,并且在其通过牛宿主的过程中可能发挥作用。在本研究中,我们研究了由rpoS编码的全局应激反应调节因子sigma(S)在耐酸性中的作用及其对大肠杆菌O157:H7在小鼠和小牛体内排出的影响。当对大肠杆菌中鉴定出的三种AR系统分别进行检测时,大肠杆菌O157:H7的rpoS突变体(rpoS::pRR10)缺乏葡萄糖抑制系统,并且精氨酸依赖性和谷氨酸依赖性AR系统的水平均降低。以10(1)至10(4) CFU的剂量将rpoS突变体和野生型菌株(ATCC 43895)接种到ICR小鼠体内后,我们发现,在给予较低剂量(10(2) CFU对10(3) CFU)的小鼠粪便中,野生型菌株的检出率更高(80%对13%)。rpoS突变体排出率的降低是由于耐酸性降低所致,因为在0.05 M磷酸盐缓冲液中接种突变体促进了其排出,并且在10(4) CFU剂量下,粪便中的回收率从27%提高到了67%。对接种了野生型菌株和rpoS突变体等量混合物的小牛粪便中的大肠杆菌O157:H7进行计数,结果表明突变体的排出量比野生型数量低10至100倍。野生型菌株和rpoS突变体之间的这种排出差异具有统计学意义(P≤0.05)。因此,sigma(S)似乎在大肠杆菌O157:H7在小鼠体内的通过以及从小牛体内的排出过程中发挥作用,可能是通过诱导葡萄糖抑制的RpoS依赖性AR决定因素的表达,从而增加对胃肠道应激的抵抗力。这些发现可能为未来旨在减少或消除牛群中这种病原体的努力提供线索。

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