IncH-Type Plasmid HarboringblaCTX-M-15,blaDHA-1, andqnrB4Genes Recovered from Animal Isolates

ABSTRACTThe whole sequence of plasmid pENVA carrying the extended-spectrum β-lactamase geneblaCTX-M-15was determined. It was identified from a series of clonally relatedKlebsiella pneumoniaesequence type 274 strains recovered from companion animals. This plasmid was 253,984 bp in size and harbored, in addition toblaCTX-M-15, a large array of genes encoding resistance to many antibiotic molecules, including β-lactams (blaTEM-1,blaDHA-1), aminoglycosides (aacA2,aadA1), tetracycline (tetA), quinolones (qnrB4), trimethoprim (dfrA15), and sulfonamides (two copies ofsul1). In addition, genes encoding resistance to mercury, tellurium, nickel, and quaternary compounds were identified. It also carried genes encoding DNA damage protection and mutagenesis repair and a locus for a CRISPR system, which corresponds to an immune system involved in protection against bacteriophages and plasmids. Comparative analysis of the plasmid scaffold showed that it possessed a structure similar to that of only a single plasmid, which was pNDM-MAR encoding the carbapenemase NDM-1 and identified from humanK. pneumoniaeisolates. Both plasmids possessed two replicons, namely, those of IncFIB-like and IncHIB-like plasmids, which were significantly different from those previously characterized. TheblaCTX-M-15gene, together with the other antibiotic resistance genes, was part of a large module likely acquired through a transposition process. We characterized here a new plasmid type carrying theblaCTX-M-15gene identified in aK. pneumoniaeisolate of animal origin. The extent to which this plasmid type may spread efficiently and possibly further enhance the dissemination ofblaCTX-M-15among animal and human isolates remains to be determined.

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Extensive Genetic Commonality among Wildlife, Wastewater, Community, and Nosocomial Isolates of Escherichia coli Sequence Type 131 (H30R1 and H30Rx Subclones) That Carry blaCTX-M-27 or blaCTX-M-15

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00519-18 ◽

2018 ◽

Vol 62 (10) ◽

Cited By ~ 16

Author(s):

Ivana Jamborova ◽

Brian D. Johnston ◽

Ivo Papousek ◽

Katerina Kachlikova ◽

Lenka Micenkova ◽

...

Keyword(s):

Escherichia Coli ◽

Companion Animals ◽

Antibiotic Resistance Genes ◽

Sequence Type ◽

Ecological Niches ◽

Beta Lactamase ◽

Class 1 Integron ◽

Content Type ◽

Extended Spectrum Beta Lactamase ◽

Genotypic Characteristics

ABSTRACT Escherichia coli sequence type 131 (ST131) is currently one of the leading causes of multidrug-resistant extraintestinal infections globally. Here, we analyzed the phenotypic and genotypic characteristics of 169 ST131 isolates from various sources (wildlife, wastewater, companion animals, community, and hospitals) to determine whether wildlife and the environment share similar strains with humans, supporting transmission of ST131 between different ecological niches. Susceptibility to 32 antimicrobials was tested by disc diffusion and broth microdilution. Antibiotic resistance genes, integrons, plasmid replicons, 52 virulence genes, and fimH-based subtypes were detected by PCR and DNA sequencing. Genomic relatedness was determined by pulsed-field gel electrophoresis (PFGE). The genetic context and plasmid versus chromosomal location of extended-spectrum beta-lactamase and AmpC beta-lactamase genes was determined by PCR and probe hybridization, respectively. The 169 ST131 study isolates segregated predominantly into blaCTX-M-15 H30Rx (60%) and blaCTX-M-27 H30R1 (25%) subclones. Within each subclone, isolates from different source groups were categorized into distinct PFGE clusters; genotypic characteristics were fairly well conserved within each major PFGE cluster. Irrespective of source, the blaCTX-M-15 H30Rx isolates typically exhibited virotype A (89%), an F2:A1:B− replicon (84%), and a 1.7-kb class 1 integron (92%) and had diverse structures upstream of the blaCTX-M region. In contrast, the blaCTX-M-27 H30R1 isolates typically exhibited virotype C (86%), an F1:A2:B20 replicon (76%), and a conserved IS26-ΔISEcp1-blaCTX-M-like structure. Despite considerable overall genetic diversity, our data demonstrate significant commonality between E. coli ST131 isolates from diverse environments, supporting transmission between different sources, including humans, environment, and wildlife.

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Third-Generation-Cephalosporin-Resistant Klebsiella pneumoniae Isolates from Humans and Companion Animals in Switzerland: Spread of a DHA-Producing Sequence Type 11 Clone in a Veterinary Setting

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.04408-14 ◽

2015 ◽

Vol 59 (5) ◽

pp. 2949-2955 ◽

Cited By ~ 19

Author(s):

Nadia Wohlwend ◽

Andrea Endimiani ◽

Thierry Francey ◽

Vincent Perreten

Keyword(s):

Klebsiella Pneumoniae ◽

Companion Animals ◽

Generation Cephalosporin ◽

Sequence Type ◽

Third Generation ◽

Content Type ◽

Human Infections ◽

Veterinary Hospital ◽

Animal Isolates

ABSTRACTCharacterization of third-generation-cephalosporin-resistantKlebsiella pneumoniaeisolates originating mainly from one human hospital (n= 22) and one companion animal hospital (n= 25) in Bern (Switzerland) revealed the absence of epidemiological links between human and animal isolates. Human infections were not associated with the spread of any specific clone, while the majority of animal infections were due toK. pneumoniaesequence type 11 isolates producing plasmidic DHA AmpC. This clonal dissemination within the veterinary hospital emphasizes the need for effective infection control practices.

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Susceptibility of Methicillin-Resistant and -SusceptibleStaphylococcus aureusIsolates of Various Clonal Lineages from Germany to Eight Biocides

Applied and Environmental Microbiology ◽

10.1128/aem.00799-18 ◽

2018 ◽

Vol 84 (13) ◽

Cited By ~ 2

Author(s):

Isa Adriana Kernberger-Fischer ◽

Carsten Krischek ◽

Birgit Strommenger ◽

Ulrike Fiegen ◽

Martin Beyerbach ◽

...

Keyword(s):

Heavy Metal ◽

Metal Tolerance ◽

Heavy Metal Tolerance ◽

Sequence Type ◽

Animal Origin ◽

Methicillin Resistant ◽

Content Type ◽

Benzethonium Chloride ◽

Broth Microdilution Method ◽

Animal Isolates

ABSTRACTFew studies have been conducted on the susceptibility of bacteria to biocides. A total of 182 methicillin-resistant and -susceptibleStaphylococcus aureusisolates collected from healthy or diseased humans and animals in Germany were included in the present study. Sixty-three isolates of animal origin and 119 human isolates were tested for their MICs to eight biocides or heavy metals by the broth microdilution method. The MIC50and MIC90values of human and animal isolates were equal or differed by not more than 1 dilution step, and statistical analysis revealed that differences between MICs of human and animal isolates were not significant. However, when taking into account the multilocus sequence type (MLST), a strong tendency (P= 0.054) to higher MICs of silver nitrate was detected for clonal complex 398 (CC398) isolates from humans compared to those from animals. Furthermore, a comparison of MIC values from isolates belonging to different clonal lineages revealed that important human lineages such as CC22 and CC5 exhibited significantly (P< 0.05) higher MICs for the biocides chlorhexidine, benzethonium chloride, and acriflavine than the main animal lineage sequence type 398 (ST398). Isolates with elevated MIC values were tested for the presence of biocide and heavy metal tolerance-mediating genes by PCR assays, and the following genes were detected:mepA(n[no. of isolates containing the gene] = 44),lmrS(n= 36),norA(n= 35),sepA(n= 22),mco(n= 5),czrC(n= 3),smr(n= 2),copA(n= 1),qacAand/or -B(n= 1),qacG(n= 2), andqacJ(n= 1). However, only for some compounds was a correlation between the presence of a biocide tolerance gene and the level of MIC values detected.IMPORTANCEBiocides play an essential role in controlling the growth of microorganisms and the dissemination of nosocomial pathogens. In this study, we determined the susceptibility of methicillin-resistant and -susceptibleS. aureusisolates from humans and animals to various biocides and heavy metal ions and analyzed differences in susceptibilities between important clonal lineages. In addition, the presence of biocide or heavy metal tolerance-mediating genes was investigated. We demonstrated that important human lineages such as CC22 and CC5 had significantly higher MIC values for chlorhexidine, benzethonium chloride, and acriflavine than the main farm animal lineage, ST398. In addition, it was shown that for some combinations of biocides and tolerance genes, significantly higher MICs were detected for carriers. These findings provide new insights intoS. aureusbiocide and heavy metal tolerance.

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Molecular Epidemiology of NDM-1-Producing Enterobacteriaceae and Acinetobacter baumannii Isolates from Pakistan

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02425-14 ◽

2014 ◽

Vol 58 (9) ◽

pp. 5589-5593 ◽

Cited By ~ 25

Author(s):

Anna L. Sartor ◽

Muhammad W. Raza ◽

Shahid A. Abbasi ◽

Kathryn M. Day ◽

John D. Perry ◽

...

Keyword(s):

Escherichia Coli ◽

Molecular Epidemiology ◽

Genetic Relatedness ◽

Antibiotic Resistance Genes ◽

Enterobacter Cloacae ◽

Content Type ◽

Plasmid Replicon ◽

Genes Encoding ◽

Clonal Spread ◽

Encoding Genes

ABSTRACTThe molecular epidemiology of 66 NDM-producing isolates from 2 Pakistani hospitals was investigated, with their genetic relatedness determined using repetitive sequence-based PCR (Rep-PCR). PCR-based replicon typing and screening for antibiotic resistance genes encoding carbapenemases, other β-lactamases, and 16S methylases were also performed. Rep-PCR suggested a clonal spread ofEnterobacter cloacaeandEscherichia coli. A number of plasmid replicon types were identified, with the incompatibility A/C group (IncA/C) being the most common (78%). 16S methylase-encoding genes were coharbored in 81% of NDM-producingEnterobacteriaceae.

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Characterization of a Multiresistant Mosaic Plasmid from a Fish Farm Sediment Exiguobacterium sp. Isolate Reveals Aggregation of Functional Clinic-Associated Antibiotic Resistance Genes

Applied and Environmental Microbiology ◽

10.1128/aem.03257-13 ◽

2013 ◽

Vol 80 (4) ◽

pp. 1482-1488 ◽

Cited By ~ 9

Author(s):

Jing Yang ◽

Chao Wang ◽

Jinyu Wu ◽

Li Liu ◽

Gang Zhang ◽

...

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Mobile Elements ◽

Fish Farms ◽

Significant Similarity ◽

Content Type ◽

Genes Encoding ◽

The Mean

ABSTRACTThe genusExiguobacteriumcan adapt readily to, and survive in, diverse environments. Our study demonstrated thatExiguobacteriumsp. strain S3-2, isolated from marine sediment, is resistant to five antibiotics. The plasmid pMC1 in this strain carries seven putative resistance genes. We functionally characterized these resistance genes inEscherichia coli, and genes encoding dihydrofolate reductase and macrolide phosphotransferase were considered novel resistance genes based on their low similarities to known resistance genes. The plasmid G+C content distribution was highly heterogeneous. Only the G+C content of one block, which shared significant similarity with a plasmid fromExiguobacterium arabatum, fit well with the mean G+C content of the host. The remainder of the plasmid was composed of mobile elements with a markedly lower G+C ratio than the host. Interestingly, five mobile elements located on pMC1 showed significant similarities to sequences found in pathogens. Our data provided an example of the link between resistance genes in strains from the environment and the clinic and revealed the aggregation of antibiotic resistance genes in bacteria isolated from fish farms.

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Comparative Genome Analysis and Global Phylogeny of the Toxin Variant Clostridium difficile PCR Ribotype 017 Reveals the Evolution of Two Independent Sublineages

Journal of Clinical Microbiology ◽

10.1128/jcm.01296-16 ◽

2016 ◽

Vol 55 (3) ◽

pp. 865-876 ◽

Cited By ~ 29

Author(s):

M. D. Cairns ◽

M. D. Preston ◽

C. L. Hall ◽

D. N. Gerding ◽

P. M. Hawkey ◽

...

Keyword(s):

Clostridium Difficile ◽

Antibiotic Resistance Genes ◽

Binary Toxin ◽

Human Origins ◽

Comparative Genome Analysis ◽

Toxin B ◽

Content Type ◽

Global Spread ◽

Animal Isolates ◽

Evolutionary Lineages

ABSTRACT The diarrheal pathogen Clostridium difficile consists of at least six distinct evolutionary lineages. The RT017 lineage is anomalous, as strains only express toxin B, compared to strains from other lineages that produce toxins A and B and, occasionally, binary toxin. Historically, RT017 initially was reported in Asia but now has been reported worldwide. We used whole-genome sequencing and phylogenetic analysis to investigate the patterns of global spread and population structure of 277 RT017 isolates from animal and human origins from six continents, isolated between 1990 and 2013. We reveal two distinct evenly split sublineages (SL1 and SL2) of C. difficile RT017 that contain multiple independent clonal expansions. All 24 animal isolates were contained within SL1 along with human isolates, suggesting potential transmission between animals and humans. Genetic analyses revealed an overrepresentation of antibiotic resistance genes. Phylogeographic analyses show a North American origin for RT017, as has been found for the recently emerged epidemic RT027 lineage. Despite having only one toxin, RT017 strains have evolved in parallel from at least two independent sources and can readily transmit between continents.

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Studies on Bd0934 and Bd3507, Two Secreted Nucleases from Bdellovibrio bacteriovorus, Reveal Sequential Release of Nucleases during the Predatory Cycle

Journal of Bacteriology ◽

10.1128/jb.00150-20 ◽

2020 ◽

Vol 202 (18) ◽

Author(s):

Ewa Bukowska-Faniband ◽

Tilde Andersson ◽

Rolf Lood

Keyword(s):

Antibiotic Resistance ◽

Life Cycle ◽

Antibiotic Resistance Genes ◽

Human Pathogens ◽

Temporal Expression ◽

Bdellovibrio Bacteriovorus ◽

Exogenous Dna ◽

Content Type ◽

Genes Encoding ◽

Predatory Bacteria

ABSTRACT Bdellovibrio bacteriovorus is an obligate predatory bacterium that invades and kills a broad range of Gram-negative prey cells, including human pathogens. Its potential therapeutic application has been the subject of increased research interest in recent years. However, an improved understanding of the fundamental molecular aspects of the predatory life cycle is crucial for developing this bacterium as a “living antibiotic.” During intracellular growth, B. bacteriovorus secretes an arsenal of hydrolases, which digest the content of the host cell to provide growth nutrients for the predator, e.g., prey DNA is completely degraded by the nucleases. Here, we have, on a genetic and molecular level, characterized two secreted DNases from B. bacteriovorus, Bd0934 and Bd3507, and determined the temporal expression profile of other putative secreted nucleases. We conclude that Bd0934 and Bd3507 are likely a part of the predatosome but are not essential for the predation, host-independent growth, prey biofilm degradation, and self-biofilm formation. The detailed temporal expression analysis of genes encoding secreted nucleases revealed that these enzymes are produced in a sequential orchestrated manner. This work contributes to our understanding of the sequential breakdown of the prey nucleic acid by the nucleases secreted during the predatory life cycle of B. bacteriovorus. IMPORTANCE Antibiotic resistance is a major global concern with few available new means to combat it. From a therapeutic perspective, predatory bacteria constitute an interesting tool. They not only eliminate the pathogen but also reduce the overall pool of antibiotic resistance genes through secretion of nucleases and complete degradation of exogenous DNA. Molecular knowledge of how these secreted DNases act will give us further insight into how antibiotic resistance, and the spread thereof, can be limited through the action of predatory bacteria.

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Population Structure and Antimicrobial Resistance of Canine UropathogenicEscherichia coli

Journal of Clinical Microbiology ◽

10.1128/jcm.00788-18 ◽

2018 ◽

Vol 56 (9) ◽

Cited By ~ 12

Author(s):

Tessa E. LeCuyer ◽

Barbara A. Byrne ◽

Joshua B. Daniels ◽

Dubraska V. Diaz-Campos ◽

G. Kenitra Hammac ◽

...

Keyword(s):

Population Structure ◽

Antimicrobial Resistance ◽

Companion Animals ◽

Sequence Type ◽

Content Type ◽

E Coli ◽

Extraintestinal Disease ◽

Human Infections ◽

Sequence Types

ABSTRACTEscherichia coliis the most common cause of human and canine urinary tract infection (UTI). Clonal groups, often with high levels of antimicrobial resistance, are a major component of theE. colipopulation that causes human UTI. While little is known about the population structure ofE. colithat causes UTI in dogs, there is evidence that dogs and humans can share fecal strains ofE. coliand that human-associated strains can cause disease in dogs. In order to better characterize theE. colistrains that cause canine UTI, we analyzed 295E. coliisolates obtained from canine urine samples from five veterinary diagnostic laboratories and analyzed their multilocus sequence types, phenotypic and genotypic antimicrobial resistance profiles, and virulence-associated gene repertoires. Sequence type 372 (ST372), an infrequent human pathogen, was the predominant sequence type in dogs at all locations. Extended-spectrum β-lactamase-producing isolates withblaCTX-Mgenes were uncommon in canine isolates but when present were often associated with sequence types that have been described in human infections. This provides support for occasional cross-host-species sharing of strains that cause extraintestinal disease and highlights the importance of understanding the role of companion animals in the overall transmission patterns of extraintestinal pathogenicE. coli.

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Tn6350, a Novel Transposon Carrying Pyocin S8 Genes Encoding a Bacteriocin with Activity against Carbapenemase-Producing Pseudomonas aeruginosa

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00100-17 ◽

2017 ◽

Vol 61 (5) ◽

Cited By ~ 2

Author(s):

Helena Turano ◽

Fernando Gomes ◽

Gesiele A. Barros-Carvalho ◽

Ralf Lopes ◽

Louise Cerdeira ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Type A ◽

Sequence Type ◽

Open Reading Frames ◽

Hypothetical Proteins ◽

Immunity Protein ◽

Content Type ◽

Commensal Strain ◽

Genes Encoding ◽

Reading Frames

ABSTRACT A novel transposon belonging to the Tn3-like family was identified on the chromosome of a commensal strain of Pseudomonas aeruginosa sequence type 2343 (ET02). Tn6350 is 7,367 bp long and harbors eight open reading frames (ORFs), an ATPase (IS481 family), a transposase (DDE catalytic type), a Tn3 resolvase, three hypothetical proteins, and genes encoding the new pyocin S8 with its immunity protein. We show that pyocin S8 displays activity against carbapenemase-producing P. aeruginosa, including IMP-1, SPM-1, VIM-1, GES-5, and KPC-2 producers.

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Enterobacter cloacae Complex Sequence Type 171 Isolates Expressing KPC-4 Carbapenemase Recovered from Canine Patients in Ohio

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01161-18 ◽

2018 ◽

Vol 62 (12) ◽

Cited By ~ 2

Author(s):

Joshua B. Daniels ◽

Liang Chen ◽

Susan V. Grooters ◽

Dixie F. Mollenkopf ◽

Dimitria A. Mathys ◽

...

Keyword(s):

Whole Genome Sequencing ◽

Genome Sequencing ◽

Companion Animals ◽

Enterobacter Cloacae ◽

Sequence Type ◽

Resistant Bacteria ◽

Whole Genome ◽

Content Type ◽

Complex Sequence ◽

Enterobacter Cloacae Complex

ABSTRACT Companion animals are likely relevant in the transmission of antimicrobial-resistant bacteria. Enterobacter xiangfangensis sequence type 171 (ST171), a clone that has been implicated in clusters of infections in humans, was isolated from two dogs with clinical disease in Ohio. The canine isolates contained IncHI2 plasmids encoding blaKPC-4. Whole-genome sequencing was used to put the canine isolates in phylogenetic context with available human ST171 sequences, as well as to characterize their blaKPC-4 plasmids.

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