Genetic analysis of shiga‐toxigenic <i>Escherichia coli</i> isolates from cattle in a limited region

Otawa, Kenichi; Sato, Masaaki; Sasaki, Takayoshi; Sasaki, Hiraku; Nonaka, Jun; Ito, Kikuji; Kuroki, Toshio; Nakai, Yutaka

doi:10.1111/j.1740-0929.2004.00185.x

Cited by 5 publications

(9 citation statements)

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“…faecium (NBRC100485 = ATCC19434, NBRC3535, NBRC100487, NBRC100488 and NBRC100602); six strains of Enterococcus faecalis (NBRC100480 = ATCC19433, NBRC3989 = ATCC23655, NBRC12964 = ATCC10100, NBRC12965 = ATCC12953, NBRC12966 = ATCC13398 and NBRC12967 = ATCC13399); six Escherichia coli strains (ATCC43888 [O157:H7, Shiga toxin gene 1: stx 1 −, gene 2: stx 2 −], [ stx 1 − , stx 2 −], H‐59 [ stx 1 +, stx 2 + ] (Otawa et al . ), D‐3b [O124:H19, stx 1 − , stx 2 + ] (Otawa et al . ), I‐7a [OUT:H21, stx 1 − , stx 2 + ] (Otawa et al .…”

Section: Methodsmentioning

confidence: 99%

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Bacteriophage control of vancomycin-resistant enterococci in cattle compost

et al. 2012

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Aims To isolate bacteriophage that infects vancomycin‐resistant enterococci (VRE) and to investigate the ability of this phage to diminish VRE number in vitro and in experimentally VRE‐inoculated compost. Methods and Results We sampled 106 solid or water samples, including 101 bovine faecal samples; lytic phage named Vrep‐5 was isolated from one bovine faecal sample by plaque assay using the clinical VRE isolate FN1 (Enterococcus faecium). Vrep‐5 generated clear plaques 1 mm in diameter and exhibited characteristics of the family Myoviridae A1, with a spherical head (122 ± 16 nm) and a contractile tail (152 ± 17 nm long). Vrep‐5 lysed other bacterial strains, including Enterococcus faecalis. Inoculation of vrep‐5 into 0·5 g unsterilized compost experimentally inoculated with FN1 at the multiplicity of infection of 1500 (8·8 × 104 CFU g−1 VRE and 1·3 × 108 PFU g−1 vrep‐5) led to a decrease of >3 log10 in VRE abundance compared with the untreated control after 24 h of incubation. Conclusions The data show that bacteriophage vrep‐5 is effective in the rapid reduction in VRE colonization in compost. Significance and Impact of the Study The present study gives valuable new knowledge in the fight against VRE in the animal production.

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Section: Methodsmentioning

confidence: 99%

“…), D‐3b [O124:H19, stx 1 − , stx 2 + ] (Otawa et al . ), I‐7a [OUT:H21, stx 1 − , stx 2 + ] (Otawa et al . ), B317a [OUT:NM, stx 1 +, stx 2 + ] (Otawa et al .…”

Section: Methodsmentioning

confidence: 99%

Bacteriophage control of vancomycin-resistant enterococci in cattle compost

et al. 2012

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“…The distribution and the origin of Shiga toxigenic genes including stx(1), stx(2) and eaeA gene in toxigenic but in non-toxigenic E. coli O157 and O157:H7 isolated from faeces and carcass of cattle could be determined by multiplex PCR (42). The mPCR analysis shows that stx(2) gene is more frequently found than that of stx(1) in STEC strains (33). Similar findings are reported by different researchers by various countries (1,7,28,31,43).…”

Section: Discussionmentioning

confidence: 99%

“…mPCR was used for the detection of stx(1) and stx(2) genes (33). The PCR mixture contained 5 μl of 10x PCR buffer, 2.5 mM MgCl 2 , 0.2 mM from each of dNTPs, 0.25 mM from each primer, 2U Taq DNA polymerase (MBI Fermentas, Lithunai), 2 μl target DNA and the final volume of 50 μl was adjusted by the addition of deionized water.…”

Section: Methodsmentioning

confidence: 99%

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Virulence genes of Shiga toxin-producing Escherichia coli O157:H7 strains isolated from calves and cattle

;ŞEKER¹

2011

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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Summary:The aim of this study was to detect E. coli O157:H7 serotype in the faeces samples collected from calves and cattle farms located of Afyonkarahisar province in Turkey and to determine the stx(1) (Shiga toxin 1) and stx(2) (Shiga toxin 2) virulence genes in the strains of E.coli O157:H7 by multiplex PCR (mPCR). In this study, E. coli O157:H7 was isolated from the 3.1% (14 out 457 faeces samples) of the calves and cattle examined, in particular 2.3% of the cattle, 2.6% of non-diarrhoeic calves and 10.6% of diarrhoeic calves were positive for E. coli O157:H7. The stx(1) and stx(2) genes were detected in 6 out of 14 (42.8%) DNA samples extracted from STEC O157:H7 strains. This study demonstrated of E. coli O157:H7 serotype in cattle and calves, which represent an important reservoir for strains that a potential risk for human infections.

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Detection of Escherichia coli in a cattle manure composting process by selective cultivation and colony polymerase chain reaction

Asano¹,

Kubori²,

Ozutsumi³

et al. 2011

J. of Env. Sc. & Hlth., Part B

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Livestock manure is suitable for use as a composting material. However, various intestinal microbes, such as Escherichia coli, are significant components of such manures. Thus, it is desirable that the level of intestinal microbes, and particularly opportunistic pathogens, in compost is inspected and counted regularly. The sensitivity and specificity of detection of E. coli in compost have been improved by selective cultivation followed by colony polymerase chain reaction (PCR) using the ECO primer. Indeed, the sensitivity of this method is higher than that of DNA extraction from compost and PCR. In this study, changes in numbers of E. coli present in a field-scale composting process over time was assessed using selective cultivation and colony PCR. Numbers of ECO-positive colonies after 24 h decreased, with a concomitant rise in compost temperature. ECO-positive colonies were not detected from 33 to 48 h. However, ECO-positive colony numbers increased beginning on day 4 and continuing until day 42. Thus, it seems likely that the high temperatures reached during the composting process did not affect E. coli numbers in the final compost. Additionally, selective cultivation followed by colony PCR using specific primers is an appropriate method of determining levels of cultivable pathogens in composted materials.

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Genetic analysis of shiga‐toxigenic Escherichia coli isolates from cattle in a limited region

Cited by 5 publications

References 35 publications

Bacteriophage control of vancomycin-resistant enterococci in cattle compost

Bacteriophage control of vancomycin-resistant enterococci in cattle compost

Virulence genes of Shiga toxin-producing Escherichia coli O157:H7 strains isolated from calves and cattle

Detection of Escherichia coli in a cattle manure composting process by selective cultivation and colony polymerase chain reaction

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