The complete DNA sequence and analysis of the virulence plasmid and of five additional plasmids carried by Shiga toxin-producing Escherichia coli O26:H11 strain H30☆

Fratamico, Pina M.; Yan, Xianghe; Caprioli, Alfredo; Esposito, Giuseppina; Needleman, David S.; Pepe, Tiziana; Tozzoli, Rosangela; Cortesi, Maria Luisa; Morabito, Stefano

doi:10.1016/j.ijmm.2010.09.002

Cited by 34 publications

(27 citation statements)

References 61 publications

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“…The presence of tet (A) of Gram-negative origin in all clones of the PCMA-3D library, and the homology of their end-read sequences to enterobacterial plasmids such as pO26-Vir, pC59-153, and pYDC637, suggests that the family Enterobacteriaceae (and E. coli in particular) are the majority tetracycline-resistant population at day 3. Further, tet (A) has been described in multi-resistant E. coli isolates from human and food animals (Fratamico et al, 2011; Wang et al, 2014; Lee et al, 2015). At day 3, tet (M) and tet (S) of Gram-positive origin were the only tetracycline resistance genes found among the clones from LAB (on MRS).…”

Section: Discussionmentioning

confidence: 99%

A Functional Metagenomic Analysis of Tetracycline Resistance in Cheese Bacteria

2017

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Metagenomic techniques have been successfully used to monitor antibiotic resistance genes in environmental, animal and human ecosystems. However, despite the claim that the food chain plays a key role in the spread of antibiotic resistance, metagenomic analysis has scarcely been used to investigate food systems. The present work reports a functional metagenomic analysis of the prevalence and evolution of tetracycline resistance determinants in a raw-milk, blue-veined cheese during manufacturing and ripening. For this, the same cheese batch was sampled and analyzed on days 3 and 60 of manufacture. Samples were diluted and grown in the presence of tetracycline on plate count milk agar (PCMA) (non-selective) and de Man Rogosa and Sharpe (MRS) agar (selective for lactic acid bacteria, LAB). DNA from the cultured bacteria was then isolated and used to construct four fosmid libraries, named after the medium and sampling time: PCMA-3D, PCMA-60D, MRS-3D, and MRS-60D. Clones in the libraries were subjected to restriction enzyme analysis, PCR amplification, and sequencing. Among the 300 fosmid clones analyzed, 268 different EcoRI restriction profiles were encountered. Sequence homology of their extremes clustered the clones into 47 groups. Representative clones of all groups were then screened for the presence of tetracycline resistance genes by PCR, targeting well-recognized genes coding for ribosomal protection proteins and efflux pumps. A single tetracycline resistance gene was detected in each of the clones, with four such resistance genes identified in total: tet(A), tet(L), tet(M), and tet(S). tet(A) was the only gene identified in the PCMA-3D library, and tet(L) the only one identified in the PCMA-60D and MRS-60D libraries. tet(M) and tet(S) were both detected in the MRS-3D library and in similar numbers. Six representative clones of the libraries were sequenced and analyzed. Long segments of all clones but one showed extensive homology to plasmids from Gram-positive and Gram-negative bacteria. tet(A) was found within a sequence showing strong similarity to plasmids pMAK2 and pO26-Vir from Salmonella enterica and Escherichia coli, respectively. All other genes were embedded in, or near to, sequences homologous to those of LAB species. These findings strongly suggest an evolution of tetracycline resistance gene types during cheese ripening, which might reflect the succession of the microbial populations. The location of the tetracycline resistance genes in plasmids, surrounded or directly flanked by open reading frames encoding transposases, invertases or mobilization proteins, suggests they might have a strong capacity for transference. Raw-milk cheeses should therefore be considered reservoirs of tetracycline resistance genes that might be horizontally transferred.

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Section: Discussionmentioning

confidence: 99%

A Functional Metagenomic Analysis of Tetracycline Resistance in Cheese Bacteria

2017

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“…The hemolysin gene, hlyA, was present in 30% of our strains, while the etpD and katP genes were absent in all isolates, despite all 3 genes being carried on pO157. Several studies have shown that these 3 genes are almost always present in O157:H7 but occur sporadically in non-O157 strains (22). Indeed, the etpD gene is rarely detected in bovine STEC (22).…”

Section: Discussionmentioning

confidence: 99%

“…Several studies have shown that these 3 genes are almost always present in O157:H7 but occur sporadically in non-O157 strains (22). Indeed, the etpD gene is rarely detected in bovine STEC (22). All of the O113:H4 (the most prevalent serotype) isolates were eaeA negative, a characteristic particularly associated with the O113 serogroup (57).…”

Section: Discussionmentioning

confidence: 99%

Serotypes and Virulence Profiles of Non-O157 Shiga Toxin-Producing Escherichia coli Isolates from Bovine Farms

Monaghan

Byrne

Fanning

et al. 2011

Appl Environ Microbiol

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Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are clinically significant food-borne pathogens. However, there is a dearth of information on serotype prevalence and virulence gene distribution, data essential for the development of public health protection monitoring and control activities for the meat and dairy industries. Thus, the objective of this study was to examine the prevalence of non-O157 STEC on beef and dairy farms and to characterize the isolates in terms of serotype and virulence markers. Bovine fecal samples (n ‫؍‬ 1,200) and farm soil samples (n ‫؍‬ 600) were collected from 20 farms throughout Ireland over a 12-month period. Shiga toxin-positive samples were cultured and colonies examined for the presence of stx 1 and/or stx 2 genes by PCR. Positive isolates were serotyped and examined for a range of virulence factors, including eaeA, hlyA, tir, espA, espB, katP, espP, etpD, saa, sab, toxB, iha, lpfA O157/OI-141 , lpfA O113 , and lpfA O157/OI-154 . Shiga toxin and intimin genes were further examined for known variants. Significant numbers of fecal (40%) and soil (27%) samples were stx positive, with a surge observed in late summer-early autumn. One hundred seven STEC isolates were recovered, representing 17 serotypes. O26:H11 and O145:H28 were the most clinically significant, with O113:H4 being the most frequently isolated. However, O2:H27, O13/O15:H2, and ONT:H27 also carried stx 1 and/or stx 2 and eaeA and may be emerging pathogens.

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“…SFO157:NM strains carry a significantly larger virulence plasmid, pSFO157, which lacks katP, toxB, and espP but carries a locus for an adhesin, sfp (244). Plasmids in non-O157 STEC strains, such as serotypes O26, O103, O111, and O145 have gene contents similar to that of pO157, with some variances (35,245,246). Interestingly, an exceptionally large plasmid, pO26-Vir, from an O26:H11 isolate, also carries a cluster that encodes a type IV pilus (245).…”

Section: Pathogenesismentioning

confidence: 99%

Recent Advances in Understanding Enteric Pathogenic Escherichia coli

et al. 2013

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SUMMARY Although Escherichia coli can be an innocuous resident of the gastrointestinal tract, it also has the pathogenic capacity to cause significant diarrheal and extraintestinal diseases. Pathogenic variants of E. coli (pathovars or pathotypes) cause much morbidity and mortality worldwide. Consequently, pathogenic E. coli is widely studied in humans, animals, food, and the environment. While there are many common features that these pathotypes employ to colonize the intestinal mucosa and cause disease, the course, onset, and complications vary significantly. Outbreaks are common in developed and developing countries, and they sometimes have fatal consequences. Many of these pathotypes are a major public health concern as they have low infectious doses and are transmitted through ubiquitous mediums, including food and water. The seriousness of pathogenic E. coli is exemplified by dedicated national and international surveillance programs that monitor and track outbreaks; unfortunately, this surveillance is often lacking in developing countries. While not all pathotypes carry the same public health profile, they all carry an enormous potential to cause disease and continue to present challenges to human health. This comprehensive review highlights recent advances in our understanding of the intestinal pathotypes of E. coli .

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The complete DNA sequence and analysis of the virulence plasmid and of five additional plasmids carried by Shiga toxin-producing Escherichia coli O26:H11 strain H30☆

Cited by 34 publications

References 61 publications

A Functional Metagenomic Analysis of Tetracycline Resistance in Cheese Bacteria

A Functional Metagenomic Analysis of Tetracycline Resistance in Cheese Bacteria

Serotypes and Virulence Profiles of Non-O157 Shiga Toxin-Producing Escherichia coli Isolates from Bovine Farms

Recent Advances in Understanding Enteric Pathogenic Escherichia coli

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