Temperate Bacteriophages Affect Pulsed-Field Gel Electrophoresis Patterns of
            <i>Campylobacter jejuni</i>

Barton, Connie; Ng, Lai-King; Tyler, Shaun; Clark, Clifford G.

doi:10.1128/jcm.01513-06

Cited by 40 publications

(32 citation statements)

References 26 publications

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“…PFGE is a time-consuming, labor-intensive method, and point mutations, insertions, and deletions of DNA can dramatically change banding patterns, resulting in misleading results. This has been reported for a number of bacteria, including Escherichia coli (5,28), Shigella sonnei (36), and Campylobacter jejuni (4). It is clear that bacteriophages are major contributors to genomic variation (4,5,8,15,16,18,21,28,36).…”

Section: Discussionmentioning

confidence: 77%

Impact of Pseudomonas aeruginosa Genomic Instability on the Application of Typing Methods for Chronic Cystic Fibrosis Infections

et al. 2010

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The Liverpool epidemic strain (LES) of Pseudomonas aeruginosa is widespread among cystic fibrosis (CF) patients in the United Kingdom and has emerged recently in North America. In this study, we report the analysis of 24 "anomalous" CF isolates of P. aeruginosa that produced inconsistent results with regard to either pulsed-field gel electrophoresis (PFGE) or PCR tests for the LES. We used a new typing method, the ArrayTube genotyping system, to determine that of the 24 anomalous isolates tested, 13 were confirmed as the LES. LES isolates could not be clearly distinguished from non-LES isolates by two other commonly used genetic fingerprinting tests, randomly amplified polymorphic DNA (RAPD) analysis and BOX-PCR, and varied considerably in their carriage of LES genomic islands and prophages. The genomic instability of the LES suggests that identification of this emerging transmissible strain could be a challenging task, and it questions whether discrimination is always a desirable feature of bacterial typing methods in the context of chronic CF infections.

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Section: Discussionmentioning

confidence: 77%

Impact of Pseudomonas aeruginosa Genomic Instability on the Application of Typing Methods for Chronic Cystic Fibrosis Infections

et al. 2010

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“…Such a possibility should be considered when PFGE is applied to epidemiological investigations of outbreaks caused by E. coli O26. It is noteworthy that variations in PFGE patterns caused by insertions or deletions of phage-associated DNA fragments have also been reported for Campylobacter jejuni (4).…”

Section: Discussionmentioning

confidence: 85%

Shiga Toxin Gene Loss and Transfer In Vitro and In Vivo during Enterohemorrhagic Escherichia coli O26 Infection in Humans

Bielaszewska

Prager

Köck

et al. 2007

Appl Environ Microbiol

180

168

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Escherichia coli serogroup O26 consists of enterohemorrhagic E. coli (EHEC) and atypical enteropathogenic E. coli (aEPEC). The former produces Shiga toxins (Stx), major determinants of EHEC pathogenicity, encoded by bacteriophages; the latter is Stx negative. We have isolated EHEC O26 from patient stools early in illness and aEPEC O26 from stools later in illness, and vice versa. Intrapatient EHEC and aEPEC isolates had quite similar pulsed-field gel electrophoresis (PFGE) patterns, suggesting that they might have arisen by conversion between the EHEC and aEPEC pathotypes during infection. To test this hypothesis, we asked whether EHEC O26 can lose stx genes and whether aEPEC O26 can be lysogenized with Stx-encoding phages from EHEC O26 in vitro. The stx 2 loss associated with the loss of Stx2-encoding phages occurred in 10% to 14% of colonies tested. Conversely, Stx2-and, to a lesser extent, Stx1-encoding bacteriophages from EHEC O26 lysogenized aEPEC O26 isolates, converting them to EHEC strains. In the lysogens and EHEC O26 donors, Stx2-converting bacteriophages integrated in yecE or wrbA. The loss and gain of Stx-converting bacteriophages diversifies PFGE patterns; this parallels findings of similar but not identical PFGE patterns in the intrapatient EHEC and aEPEC O26 isolates. EHEC O26 and aEPEC O26 thus exist as a dynamic system whose members undergo ephemeral interconversions via loss and gain of Stx-encoding phages to yield different pathotypes. The suggested occurrence of this process in the human intestine has diagnostic, clinical, epidemiological, and evolutionary implications.Escherichia coli serogroup O26 has members classified as enterohemorrhagic E. coli (EHEC) or atypical enteropathogenic E. coli (aEPEC). EHEC O26 strains constitute the most common non-O157 EHEC group associated with diarrhea and hemolytic uremic syndrome (HUS) in Europe (16,18,19,25,48,51). EHEC O26 is also the most common non-O157 EHEC serogroup in the United States, where, between 1983 and 2002, it accounted for 22% of non-O157 EHEC clinical isolates (10). In a recent prospective study from Montana, half of EHEC O26 isolates originated from patients with bloody diarrhea (23). Moreover, EHEC O26 has spread globally (24).EHEC O26 strains produce Shiga toxin 1 (Stx1) and Stx2, either singly or together (10, 54). Indeed, phage H19B from a clinical EHEC O26 isolate that carries stx 1 was one of the first Stx-converting phages described (45). Moreover, these strains contain the intimin-encoding eae gene (6, 54), an important characteristic of EHEC (33). EHEC O26 represents a highly dynamic group of organisms that rapidly engender new pathogenic clones (54). This is exemplified by emergence of a novel EHEC O26:H11 clonal subgroup in Germany in the 1990s that possessed stx 2 as the sole stx gene, in contrast to stx 1 , exclusively identified in EHEC O26 previously. The pathogenicity of this clone was demonstrated by its strong association with HUS (29, 54) and its ability to spread rapidly (2, 54).aEPEC O26 strains do not harbor stx ...

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“…The differences between the two PFGE subtypes are likely due to the gain or loss of a single KpnI site in the top band of the PFGE pattern. Other investigators also reported the occurrence of different PFGE variants from a single clone of C. jejuni due to genetic variation caused by recombinations, mutations, or insertions/deletions (3,16,45). Despite these subtle differences as determined by PFGE and cmp sequences, we conclude that the examined sheep isolates are clonally related.…”

Section: Discussionmentioning

confidence: 98%

Emergence of a Tetracycline-ResistantCampylobacter jejuniClone Associated with Outbreaks of Ovine Abortion in the United States

Şahin¹,

Plummer²,

Jordan³

et al. 2008

J Clin Microbiol

120

179

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Campylobacter infection is one of the major causes of ovine abortions worldwide. Historically, Campylobacter fetus subsp. fetus was the major cause of Campylobacter-associated abortion in sheep; however, Campylobacter jejuni is increasingly associated with sheep abortions. We examined the species distribution, genotypes, and antimicrobial susceptibilities of abortion-associated Campylobacter isolates obtained from multiple lambing seasons on different farms in Iowa, Idaho, South Dakota, and California. We found that C. jejuni has replaced C. fetus as the predominant Campylobacter species causing sheep abortion in the United States. Most strikingly, the vast majority (66 of 71) of the C. jejuni isolates associated with sheep abortion belong to a single genetic clone, as determined by pulsed-field gel electrophoresis, multilocus sequence typing, and cmp gene (encoding the major outer membrane protein) sequence typing. The in vitro antimicrobial susceptibilities of these isolates to the antibiotics that are routinely used in food animal production were determined using the agar dilution test. All of the 74 isolates were susceptible to tilmicosin, florfenicol, tulathromycin, and enrofloxacin, and 97% were sensitive to tylosin. However, all were resistant to tetracyclines, the only antibiotics currently approved in the United States for the treatment of Campylobacter abortion in sheep. This finding suggests that feeding tetracycline for the prevention of Campylobacter abortions is ineffective and that other antibiotics should be used for the treatment of sheep abortions in the United States. Together, these results indicate that a single tetracycline-resistant C. jejuni clone has emerged as the major cause of Campylobacter-associated sheep abortion in the United States.

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Temperate Bacteriophages Affect Pulsed-Field Gel Electrophoresis Patterns of Campylobacter jejuni

Cited by 40 publications

References 26 publications

Impact of Pseudomonas aeruginosa Genomic Instability on the Application of Typing Methods for Chronic Cystic Fibrosis Infections

Impact of Pseudomonas aeruginosa Genomic Instability on the Application of Typing Methods for Chronic Cystic Fibrosis Infections

Shiga Toxin Gene Loss and Transfer In Vitro and In Vivo during Enterohemorrhagic Escherichia coli O26 Infection in Humans

Emergence of a Tetracycline-ResistantCampylobacter jejuniClone Associated with Outbreaks of Ovine Abortion in the United States

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