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Objectives: This study aimed to compare ESBL-producing Escherichia coli causing infections in humans with infecting or commensal isolates from animals and isolates from food of animal origin in terms of the strain types, the ESBL gene present and the plasmids that carry the respective ESBL genes. Methods: A collection of 353 ESBL-positive E. coli isolates from the UK, the Netherlands and Germany were studied by MLST and ESBL genes were identified. Characterization of ESBL gene-carrying plasmids was performed using PCR-based replicon typing. Moreover, IncI1-Iγ and IncN plasmids were characterized by plasmid MLST. Results: The ESBL-producing E. coli represented 158 different STs with ST131, ST10 and ST88 being the most common. Overall, blaCTX-M-1 was the most frequently detected ESBL gene, followed by blaCTX-M-15, which was the most common ESBL gene in the human isolates. The most common plasmid replicon type overall was IncI1-Iγ followed by multiple IncF replicons. Conclusions: ESBL genes were present in a wide variety of E. coli STs. IncI1-Iγ plasmids that carried the blaCTX-M-1 gene were widely disseminated amongst STs in isolates from animals and humans, whereas other plasmids and STs appeared to be more restricted to isolates from specific hosts. [ABSTRACT FROM AUTHOR]

Objectives The characterization of CTX-M-15 β-lactamase-producing Escherichia coli and Salmonella isolates originating mainly from German livestock and food. Methods E. coli (526, mainly commensals) and Salmonella (151) non-human isolates resistant to third-generation cephalosporins, originating from routine and monitoring submissions (2003–12) to the Federal Institute for Risk Assessment and different national targeted studies (2011–12), were examined for the presence of blaCTX-M-15 genes by PCR amplification/sequencing. Additional resistance and virulence genes were screened by DNA microarray and PCR amplification. E. coli isolates with blaCTX-M-15 were characterized by phylogenetic grouping, PFGE and multilocus sequence typing (MLST). The blaCTX-M-15 plasmids were analysed by replicon typing, plasmid MLST, S1 nuclease PFGE and Southern blot hybridization experiments. Results Twenty-one E. coli (livestock, food and a toy; 4.0%) and two Salmonella (horse and swine; 1.3%) isolates were CTX-M-15 producers. E. coli isolates were mainly ascribed to three clonal lineages of sequence types ST678 (German outbreak with enteroaggregative Shiga-toxin-producing E. coli O104:H4; salmon, cucumber and a toy), ST410 (poultry, swine and cattle farms) and ST167/617 (swine farms and turkey meat). The blaCTX-M-15 genes were located on IncI1 and multireplicon IncF plasmids or on the chromosome of E. coli ST410 isolates. Conclusions The prevalence of CTX-M-15-producing isolates from non-human sources in Germany is still low. The blaCTX-M-15 gene is, however, present in multidrug-resistant E. coli clones with pathogenic potential in livestock and food. The maintenance of the blaCTX-M-15 gene due to chromosomal carriage is noteworthy. The possibility of an exchange of CTX-M-15-producing isolates or plasmids between livestock and humans (in both directions) deserves continuous surveillance. [ABSTRACT FROM PUBLISHER]

Objectives Between 20 000 and 35 000 cases of salmonellosis are detected annually in Germany, but only a few Salmonella are resistant to third-generation cephalosporins. The German National Reference Centre for Salmonella and other Enterics obtained 150 Salmonella enterica isolates from human infections between 2005 and 2011. In the present study we identified the β-lactamase genes causing resistance to third-generation cephalosporins in these isolates. Methods For all isolates serotyping and antimicrobial susceptibility testing were performed. The presence of β-lactamase genes was detected by PCR amplification and sequencing. Isolates with identical serovar and β-lactamase genes were typed by XbaI macrorestriction followed by PFGE. Broth mate conjugation assays and plasmid analysis using S1 nuclease restriction of genomic DNA and subsequent PFGE as well as PCR-based replicon typing were performed for selected isolates. Results The 150 isolates were assigned to 30 different serovars, with S. enterica serovar Typhimurium (n = 73; 48.7%) as the most prevalent. Two different AmpC β-lactamase genes (blaCMY-2, n = 8; blaACC-1, n = 6) and various extended-spectrum β-lactamase (ESBL) genes were identified. The majority harboured the blaCTX-M-1 gene (n = 91; 60.7%) followed by blaCTX-M-14 (n = 12; 8.0%) and blaSHV-12 (n = 11; 7.3%). Typing of strains and subsequent comparison with selected Salmonella isolates from livestock revealed the presence of several clones in both humans and livestock. Conclusions The wide spread of ESBL and AmpC genes in Salmonella of various serovars is most probably due to transfer of conjugative plasmids. Furthermore, our data indicate the clonal spread of distinct cephalosporin-resistant Salmonella strains from livestock to humans. [ABSTRACT FROM PUBLISHER]

Objectives To characterize a 5.9 kb aac(6′)-Ib-cr-harbouring plasmid that was detected in a clinical Salmonella Typhimurium DT104B strain. Methods Extraction and purification of plasmid DNA and electrotransformation assays were carried out in order to obtain kanamycin-resistant transformants. MICs of several fluoroquinolones and aminoglycosides were determined. DNA sequencing was performed by primer walking on purified plasmid preparations. The new plasmid nucleotide sequence was analysed and compared with available sequences using bioinformatic tools. Results pMdT1 is a 5.9 kb mobilizable ColE1-like plasmid that harbours aac(6′)-Ib-cr4, a gene encoding a new variant of the AAC(6′)-Ib-cr protein (225 amino acids). This active protein conferred resistance to tobramycin and kanamycin, and also decreased susceptibility to ciprofloxacin and norfloxacin in the transformant strain, as MICs demonstrated. The mobilization region, necessary for horizontal transfer and composed of the mobA, mobB, mobC and mobD genes, displayed a high degree of identity with those from representative ColE1-like plasmids. The basis of mobility (bom), oriT and origin of replication regions were also detected. Apart from the acetylase-encoding gene, three other open reading frames (ORFs) were determined. No similarities were found when the ORF1 sequence was compared with the sequences included in GenBank. The deduced ORF2 protein predicted a CopG-like structure characteristic of transcriptional regulators, and the deduced ORF3 protein was identical to macrophage stimulating factors. Conclusions The pMdT1 is the smallest mobilizable ColE1-like plasmid containing an aac(6′)-Ib-cr gene that has been described so far. [ABSTRACT FROM PUBLISHER]

The article offers information on a study which examined extended-spectrum β-lactamases in German Salmonella enterica isolate subsp. enterica serovar Typhimurium 4,[5],12:i:-European clone. Nine strains of German Salmonella enterica were examined for the presence of β-lactams. Study authors found that the strains share the resistance phenotype ampicillin/cefalotin/cefuroxime/ticarcillin/piperacillin/ceftiofur/ceftriaxone/cefotaxime/aztreonam/cefopodoxime.

Objectives This study was initiated to collect retrospective information on the occurrence of plasmid-mediated quinolone resistance (PMQR) in Salmonella enterica and Escherichia coli isolates in Europe and to identify the responsible genes. Methods Databases of national reference laboratories containing MIC values for Salmonella and E. coli isolated between 1994 and 2009 in animals, humans, food and the environment from 13 European countries were screened for isolates exhibiting a defined quinolone resistance phenotype, i.e. reduced susceptibility to fluoroquinolones and nalidixic acid. PCR and sequence analysis were performed to identify the responsible PMQR genes. Results Screening of databases of 13 European countries resulted in a selection of 1215 Salmonella and 333 E. coli isolates. PMQR genes were identified in 59% of the Salmonella isolates and 15% of the E. coli isolates selected. In Salmonella, qnrS1 (n = 125) and variants of qnrB (n = 138) were frequently identified, whereas qnrA1 (n = 3) and aac(6′)-1b-cr (n = 3) were rarely found. qnrD was detected in 22 Salmonella isolates obtained from humans and animals. In E. coli, qnrS1 was identified in 19 isolates and qnrB19 was found in one isolate. No qnrC or qepA genes were detected in either Salmonella or E. coli. Conclusions This study shows the occurrence and dissemination of PMQR genes in Salmonella and E. coli in Europe with a defined quinolone resistance phenotype. We also report the first detection of qnrD in Salmonella collected in Europe. [ABSTRACT FROM PUBLISHER]

Objectives To broaden knowledge of the molecular bases and genetics of multidrug resistance in clinical isolates of Salmonella enterica serotype 4,5,12:i:− belonging to the Spanish clone. Methods The relatedness of the isolates was determined by phage typing and XbaI-PFGE. Resistance genes, integrons and transposable elements were identified by PCR amplification and sequencing. Plasmids were characterized by alkaline lysis, S1-PFGE, conjugation, replicon typing and Southern blot hybridization. Results The isolates were closely related and resistant to five to seven antimicrobials (ampicillin, chloramphenicol, gentamicin, streptomycin/spectinomycin, sulphonamides, trimethoprim and tetracycline, arranged in different combinations). Most of the responsible genes were provided by a conventional class 1 integron with the dfrA12-orfF-aadA2 variable region, an atypical class 1 integron containing sul3 next to the estX-psp-aadA2-cmlA1-aadA1 variable region and a truncated Tn1721 transposon carrying tet(A). A defective Tn21 with the mer operon and ISVsa3 associated with sul2 were also detected. All resistance genes and mobile genetic elements were located on large, non-conjugative and highly variable plasmids carrying one (A/C) or two (A/C and N) replicons, as well as virulence genes of pSLT. Conclusions IncA/C plasmids are responsible for multidrug resistance in an increasing number of relevant human and animal bacterial pathogens, and hence are regarded as an important threat to public health. Those found in the Spanish clone of Salmonella 4,5,12:i:− constitute a relevant example of short-term evolution, and could have been involved in the successful adaptation of this pathogen. [ABSTRACT FROM PUBLISHER]

Objectives: In view of the intercontinental emergence of Escherichia coli clone O25:H4-ST131 producing CTX-M-15 extended-spectrum b-lactamase (ESBL) in human clinical settings it would be of great interest to explore its existence in animals to unravel a possible reservoir function and the origin and transmission of this group of multiresistant strains. Methods: A total of 177 clinical phenotypically ESBL-producing E. coli isolates, mainly obtained from companion animals with urinary tract infections, wound infections and diarrhoea, were collected in a veterinary diagnostic laboratory covering a European-wide service area. They were screened for molecular subtype O25b and multilocus sequence type 131. O25b-ST131 isolates were subsequently tested for ESBL types, and phenotypic and genotypic resistance determinants. Further characterization of the strains was performed by PFGE and virulence gene typing. Results: Ten (5.6%) of 177 phenotypically ESBL-producing E. coli isolates, nine strains from dogs and one strain from a horse, were allocated to the B2-O25b-ST131 lineage. Nine of these isolates harboured a CTX-M-15-type b-lactamase enzyme while one strain possessed an SHV-12-type ESBL. Macrorestriction analysis revealed a cluster formation of six of the animal CTX-M-15-type ESBL-producing strains from five different European countries together with a human control strain constituting a group of clonally related strains at a similarity value of 87.0%. Conclusions: Our findings demonstrate that the group of clonally related human B2-O25:H4-ST131 CTX-M-15-type ESBL-producing E. coli strains is present in companion animals from various European countries. This highlights the possibility of inter-species transmission of these multiresistant strains from human to animal and vice versa. [ABSTRACT FROM AUTHOR]

Background. Although leptospirosis is a reemerging zoonosis of global importance, outbreaks related to agricultural exposures are primarily situated in tropical countries. In July 2007, a suspected leptospirosis outbreak was recognized among strawberry harvesters from Eastern Europe who were working in Germany. An investigation was initiated to identify the outbreak source and the risk factors for infection. Methods. We conducted a retrospective cohort study with use of a questionnaire administered to harvesters by health authorities in Romania, Slovakia, and Poland. Collected serum samples were tested by microscopic agglutination test and immunoglobulin M enzyme-linked immunosorbent assay. A case patient was defined as a person who worked in the strawberry field during the period 5 June-8 September 2007 and had leptospirosiscompatible symptoms and either an antibody titer ⩾1:800 and a positive immunoglobulin M enzyme-linked immunosorbent assay result (for a confirmed case) or no serological confirmation (for a suspected case). Local rodents were examined for leptospirosis. Results. Among 153 strawberry harvesters, we detected 13 confirmed case patients who had test results positive for antibodies against Leptospira species serogroup Grippotyphosa and 11 suspected case patients (attack rate, 16%). Risk of disease increased with each day that an individual worked in the rain with hand wounds (odds ratio, 1.1; 95% confidence interval, 1.04-1.14) and accidental rodent contact (odds ratio, 4.8; 95% confidence interval, 1.5-15.9). Leptospires of the serogroup Grippotyphosa were isolated from the kidneys of 7 (64%) of 11 voles. Conclusions. This is, to our knowledge, the largest leptospirosis epidemic to occur in Germany since the 1960s. Contact between hand lesions and contaminated water or soil and infected voles was the most likely outbreak source. The unusually warm winter of 2006-2007 supported vole population growth and contributed to this resurgence of leptospirosis in Germany. Because of ongoing climate change, heightened awareness of leptospirosis in temperate regions is warranted. [ABSTRACT FROM AUTHOR]

A letter to the editor about extended-spectrum β-lactamases (ESBL) of the CTX-M-8-positive isolates of species Escherichia coli, Salmonella enterica and Enterobacter aerogenes in Garmany is presented.

A letter to the editor is presented discussing research on the New Delhi metallo-beta-lactamase-1 (NDM-1)-producing Salmonella enterica subsp. enterica serovar Corvallis isolated from a wild bird in Germany.

A letter to the editor is presented about the presence of a carbapenemase-encoding gene in Salmonella enterica isolated from poultry and pig farms.

The article examines the presence of a carbapenemase-encoding gene in Escherichia coli isolated from a livestock farm. It states that the occurrence of extended spectrum Β-lactamase (ESBL) in Enterobacteriaceae isolated from food-producing animals and the increase in their prevalence is a worldwide problem, however, the presence of carbapenemases is restricted to humans. It cites that the presence of this gene and the possibility of their transmission is a major concern for public health.

The article discusses a research study on the prevalence of plasmid-mediated quinolone resistance (PMQR) determinants in Salmonella and Escherichia isolates from reptiles in Germany. Isolates with nalixidic acid MIC and ciprofloxacin MIC were tested for the presence of PMQR-encoding genes. Plasmid profile analysis by DNA extraction showed the presence of qnrB19 gene in seven strains belonging to different Salmonella subspecies and serotypes. The study also confirmed the plasmid location of the qnr genes.

The article focuses on a study which determined the prevalence of Escherichia coli harbouring genes of the CTX-M-ESBL type on wildlife animal species with close contact with human settlements, the brown rat. Faecal samples from urban brown rats were collected in Berlin, Germany. The isolates which were identified using CHORM agar orientation and were further processed by agar disk diffusion testing as a preliminary screen for antimicrobial resistance. Antibiotics wherein phenotypic antimicrobial resistance were detected include cefatoxime and cefalotin.

The article describes the complete nucleotide sequence of the 2699 bp ColE-like quinolone resistance (qnrB19) plasmid pSGI15. It examines several mutiresistant Salmonella strains isolated from animals, food and humans between 2001 and 2006. It reveals that of the isolates studied, 53 percent were found to beSGI1 positive.

Objectives Detection and characterization of extended-spectrum β-lactamases (ESBLs) and AmpC-encoding genes was conducted in German Salmonella isolated from different sources from 2003 to 2007. Methods Non-duplicate German isolates from the National Salmonella Reference Laboratory Collection (2003–07) with ceftiofur MICs of ≥4 mg/L were tested for β-lactam/β-lactamase inhibitor susceptibility, presence of ESBLs or AmpC-encoding genes, class 1 and 2 integrons, other resistance genes, and IS26 and ISEcp1 sequences by PCR/sequencing. The isoelectric point of the β-lactamase was determined. Strains were analysed by PFGE and plasmid profiling. The bla genes were mapped by Southern-blot hybridization. Plasmids were characterized by rep-PCR typing. Results Sixteen isolates (10 Salmonella Typhimurium, 2 Salmonella Anatum, 2 Salmonella Paratyphi B dT , 1 Salmonella Infantis and 1 Salmonella London) carried blaCTX-M (15 blaCTX-M-1 and one blaCTX-M-15) genes located on self-transferable IncB/O, IncI1 and/or IncN plasmids. Seven of the Salmonella Typhimurium isolates carried the SGI1-M variant. Six isolates (five Salmonella Agona and one Salmonella Kentucky) carried the blaCMY-2 gene on IncI1 conjugative plasmids. blaTEM-20 genes were detected in two Salmonella Paratyphi B dT isolates, and blaTEM-52 in one Salmonella Paratyphi B dT and one Salmonella Virchow, located on IncI1 plasmids. All Salmonella Paratyphi isolates harboured a 2300 bp/dfrA1-sat2-aadA1 class 2 integron. Conclusions Among the 22 679 German Salmonella isolates investigated, the ESBL and AmpC β-lactamase prevalence was still low; however, it is slowly increasing. Various β-lactamase genes are linked to a variety of genetic elements capable of horizontal DNA transfer. Consequently, their dissemination is likely and demands adequate risk management strategies. [ABSTRACT FROM AUTHOR]