Limited similarity between plasmids encoding CTX-M-1 ß-lactamase in Escherichia coli from humans, pigs, cattle, organic poultry layers and horses in Denmark.

CTX-M-1 is a common extended-spectrum ß-lactamase (ESBL) in Escherichia coli from animals and is often detected among human clinical isolates. The objective of this study was to investigate the epidemiological relationship between CTX-M-1-producing E. coli isolated from patients and animals in Denmark between 2006 and 2010. In total, 65 CTX-M-1-producing isolates from patients (n=22), pigs (n=21), cattle (n=4), organic poultry layers (n=3) and horses (n=15) were typed by pulsed-field gel electrophoresis (PFGE). Plasmids harbouring blaCTX-M-1 were characterised by S1 PFGE, PCR-based replicon typing, plasmid multilocus sequence typing, restriction fragment length polymorphism, and sequencing. Human and animal strains were unrelated based on PFGE. IncI1 was more common in human isolates (13/22) than in animal isolates (7/43), whereas the opposite trend was observed for IncN (5/22 human isolates and 24/43 animal isolates). Full characterisation of the plasmids harbouring blaCTX-M-1 revealed host-specific patterns in the distribution of plasmid types, with specific IncI1, IncN and IncH1 plasmid subtypes being predominant in humans, livestock and horses, respectively. Three indistinguishable human, bovine and porcine IncI1/ST49 plasmids had high nucleotide sequence homology and differed by the presence of IS66 elements in the bovine plasmid and the absence of one gene within the microcin-encoding operon in the human plasmid. In conclusion, this work suggests a minor contribution by animals to the occurrence of CTX-M-1 in human E. coli infections in Denmark during the study period.

Complete Genome Sequence of Pediococcus pentosaceus Strain SL4.

Pediococcus pentosaceus SL4 was isolated from a Korean fermented vegetable product, kimchi. We report here the whole-genome sequence (WGS) of P. pentosaceus SL4. The genome consists of a 1.79-Mb circular chromosome (G+C content of 37.3%) and seven distinct plasmids ranging in size from 4 kb to 50 kb.

Expansion of the IncX plasmid family for improved identification and typing of novel plasmids in drug-resistant Enterobacteriaceae.

IncX plasmids are narrow host range plasmids of Enterobactericeae that have been isolated for over 50years. They are known to encode type IV fimbriae enabling their own conjugative transfer, and to provide accessory functions to their host bacteria such as resistance towards antimicrobial agents and biofilm formation. Previous plasmid-based replicon typing procedures have indicated that the prevalence of IncX plasmids is low among members of the Enterobacteriaceae. However, examination of a number of IncX-like plasmid sequences and their occurrence in various organisms suggests that IncX plasmid diversity and prevalence is underappreciated. To address these possible shortcomings, we generated additional plasmid sequences of IncX plasmids of interest and compared them to the genomes of all sequenced IncX-like plasmids. These comparisons revealed that IncX plasmids possess a highly syntenic plasmid backbone, but that they are quite divergent with respect to nucleotide and amino acid similarity. Based on phylogenetic comparisons of the sequenced IncX plasmids, the IncX plasmid group has been expanded to include at least four subtypes, IncX1-IncX4. A revised IncX plasmid replicon typing procedure, based upon these sequences and subtypes, was then developed. Use of this revised typing procedure revealed that IncX plasmid occurrence among bacterial populations is much more common than had previously been acknowledged. Thus, this revised procedure can be used to better discern the occurrence of IncX type plasmids among enterobacterial populations.

Investigation of diversity of plasmids carrying the blaTEM-52 gene.

OBJECTIVES: To investigate the diversity of plasmids that carry bla(TEM-52) genes among Escherichia coli and Salmonella enterica originating from animals, meat products and humans. METHODS: A collection of 22 bla(TEM-52)-encoding plasmids was characterized by restriction fragment length polymorphism (RFLP), replicon typing (by PCR or replicon sequencing), susceptibility testing, assessment of plasmid ability to self-transfer by conjugation and typing of the genetic environment of the bla(TEM-52) gene. Detected IncI1 plasmids underwent further plasmid multilocus sequence typing. RESULTS: RFLP profiles demonstrated dissemination of bla(TEM-52) in Denmark (imported meat from Germany), France, Belgium and the Netherlands from 2000 to 2006 by mainly two different plasmids, one encoding bla(TEM-52b) (IncX1A, 45 kb) and the other bla(TEM-52c) (IncI1, 80 kb). In addition, bla(TEM-52b) was also found to be located on various other plasmids belonging to IncA/C and IncL/M, while bla(TEM-52c) was found on IncN-like as well as on IncR plasmids. In the majority of cases (n = 21) the bla(TEM-52) gene was located on a Tn3 transposon. Seven out of 10 bla(TEM-52) plasmids tested in conjugation experiments were shown to be capable of self-transfer to a plasmid-free E. coli recipient. CONCLUSIONS: The bla(TEM-52) gene found in humans could have been transmitted on transferable plasmids originating from animal sources. Some of the bla(TEM-52) plasmids carry replicons that differ from the classical ones. Two novel replicons were detected, IncX1A and IncN-like. Unlike its predecessor bla(TEM-1), the bla(TEM-52) gene was not detected on F-type replicons suggesting that this gene evolved on other types of plasmid scaffolds.

Characterization of a small erythromycin resistance plasmid pLFE1 from the food-isolate Lactobacillus plantarum M345.

This paper reports the complete 4031 bp nucleotide sequence of the small erythromycin resistance plasmid pLFE1 isolated from the raw-milk cheese isolate Lactobacillus plantarum M345. Analysis of the sequence revealed the coding regions for the erythromycin resistance determinant Erm(B). A replication initiation protein RepB was identified belonging to the RepB proteins of the pMV158 family of rolling-circle replicating plasmids. The transcriptional repressor protein CopG and a small counter transcribed RNA, two elements typically involved in replication control within this family were also found. A putative replication initiation site including a single-strand origin (sso) -like region succeeded by a characteristic pMV158 family double-strand origin (dso) was located upstream of the replication region. An open reading frame following a typical origin of transfer (oriT) site and coding for a putative truncated mobilization (Mob) protein with a size of 83 aa was detected. The product of the putative mob gene showed large similarity to the N-terminal region of the pMV158 family of Pre/Mob proteins, but was much smaller than other proteins of this family. We therefore suggest that the Mob function in pLFE1 is supplied in trans from another plasmid present in L. plantarum M345.Filter-mating experiments showed that pLFE1 has a broad host-range with transconjugants obtained from Lactobacillus rhamnosus, Lactococcus lactis, Listeria innocua, the opportunistic pathogen Enterococcus faecalis and the pathogen Listeria monocytogenes.