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At least 577 records · Page 32Linked to original sources
PubMed2005

Tn7 distribution in Helicobacter pylori: a selective paradox.

The presence of class 1, 2 and 3 integrons in Helicobacter pylori, which in other species are usually related to antimicrobial resistance, were looked for in 40 epidemiologically-unrelated isolates. The presence of the class 2 integrase, which led to the discovery of Tn7, was found in 15 of 35 different clones identified by lspA-glmM polymorphism. MIC values for trimethoprim and streptomycin were not affected by the presence or absence of dfrA1 and aadA1 genes. The polyclonal spread of Tn7 showed the ability of H. pylori to harbour antimicrobial resistance mechanisms, usually located on plasmids, and also suggested that other important mechanisms of resistance may also be acquired, which would result in failure of chemotherapy.

Anti-Bacterial Agents↗
PubMed2005

Quinolone resistance in bacteria: emphasis on plasmid-mediated mechanisms.

Bacterial resistance to quinolones/fluoroquinolones has emerged rapidly and such resistance has traditionally been attributed to the chromosomally mediated mechanisms that alter the quinolone targets (i.e. DNA gyrase and topoisomerase IV) and/or overproduce multidrug resistance efflux pumps. However, the discovery of the plasmid-borne quinolone resistance determinant, named qnr, has substantially broadened our horizon on the molecular mechanisms of quinolone resistance. Several recent reports of Qnr or its homologues encoded by transferable plasmids in Gram-negative bacteria isolated worldwide highlight the significance of the emerging plasmid-mediated mechanism(s). This also alerts us to the potential rapid dissemination of quinolone resistance determinants. Qnr belongs to the pentapeptide repeat family and protects DNA gyrase from the action of quinolone agents including the newer fluoroquinolones. This protection interplays with chromosomal mechanisms to raise significantly the resistance levels. The qnr-bearing strains generate quinolone-resistant mutants at a much higher frequency than those qnr-free strains. Furthermore, the qnr-plasmids are integron-associated and carry multiple resistance determinants providing resistance to several classes of antimicrobials including beta-lactams and aminoglycosides. The high quinolone resistance rates in Escherichia coli are used to address issues of quinolone resistance, and possible strategies for minimising quinolone resistance are discussed.

Anti-Bacterial Agents↗
PubMed2005

Detection of sul1, sul2 and sul3 in sulphonamide resistant Escherichia coli isolates obtained from healthy humans, pork and pigs in Denmark.

The occurrence of sulphonamide resistance was investigated in 998 Escherichia coli isolates, obtained from pig faeces collected at slaughter, Danish pork collected at retail outlets and from faeces from healthy persons in Denmark. In total 18% (n=35), 20% (n=38) and 26% (n=161) of the E. coli isolates obtained from humans, pork and pigs, respectively, were resistant to sulphonamide. All sulphonamide resistant E. coli isolates were investigated for the presence of sul1, sul2, sul3 and intI1 genes by PCR. The sul1 gene was detected in 40% (n=14), 29% (n=11) and 55% (n=88) of the sulphonamide resistant isolates from humans, pork and pigs, respectively. The sul2 gene was detected in 80% (n=28), 76% (n=29) and 50% (n=81) of isolates from humans, pork and pigs, respectively. None of the human isolates were PCR-positive for sul3, whereas sul3 was present in 5% of the pork isolates and 11% of the pig isolates. Of the 113 sul1 positive isolates, 97 carried the integron-associated integrase gene intI1. All 20 sul3 positive isolates were positive for intI1, and in 12 of these isolates sul3 was the only sulphonamide resistance gene detected. The origin of sul1 and sul2 found in isolates from healthy humans is speculative, but their spread from pigs to humans via the food chain is possible.

Abattoirs↗
PubMed2006

The CTX-M beta-lactamase pandemic.

In the past decade CTX-M enzymes have become the most prevalent extended-spectrum beta-lactamases, both in nosocomial and in community settings. The insertion sequences (ISs) ISEcp1 and ISCR1 (formerly common region 1 [CR1] or orf513) appear to enable the mobilization of chromosomal beta-lactamase Kluyvera species genes, which display high homology with blaCTX-Ms. These ISs are preferentially linked to specific genes: ISEcp1 to most blaCTX-Ms, and ISCR1 to blaCTX-M-2 or blaCTX-M-9. The blaCTX-M genes embedded in class 1 integrons bearing ISCR1 are associated with different Tn402-derivatives, and often with mercury Tn21-like transposons. The blaCTX-M genes linked to ISEcp1 are often located in multidrug resistance regions containing different transposons and ISs. These structures have been located in narrow and broad host-range plasmids belonging to the same incompatibility groups as those of early antibiotic resistance plasmids. These plasmids frequently carry aminoglycoside, tetracycline, sulfonamide or fluoroquinolone resistance genes [qnr and/or aac(6')-Ib-cr], which would have facilitated the dissemination of blaCTX-M genes because of co-selection processes. In Escherichia coli, they are frequently carried in well-adapted phylogenetic groups with particular virulence-factor genotypes. Also, dissemination has been associated with different clones (CTX-M-9 or CTX-M-14 producers) or epidemic clones associated with specific enzymes such as CTX-M-15. All these events might have contributed to the current pandemic CTX-M beta-lactamase scenario.

Anti-Bacterial Agents↗
PubMed2005

SlyA regulates the collagenase-mediated cytopathic phenotype in multiresistant Salmonella.

Salmonella enterica serotype Typhimurium phagetype DT104 (DT104) is a foodborne pathogen with a multiresistant phenotype conferred by a genomic-based integron structure designated as SGI1. Recently, a novel cytopathic phenotype was ascribed to several isolates of DT104 recovered from veal calves. This phenotype is dependent upon clg, a gene encoding a collagenase in Salmonella. Using a novel transposon system and an RT-PCR assay for detection of clg expression, we identified SlyA as a regulator of the collagenase-mediated phenotype. The function of SlyA, in regards to clg expression, is to repress the synthesis of Clg. Derepression ensued in the absence of SlyA or in the presence of a truncated version of SlyA with the latter being relevant for maintenance of another virulence aspect mediated by SlyA, i.e. survival within macrophages. The SlyA-mediated effect on clg expression was restricted to DT104 and other Salmonella phagetypes and serotypes possessing SGI1 thus suggesting co-regulation by an SGI1-specific component.

Amino Acid Sequence↗
PubMed2005

Microarray-based detection of genetic heterogeneity, antimicrobial resistance, and the viable but nonculturable state in human pathogenic Vibrio spp.

The morbidity and mortality associated with Vibrio-mediated waterborne diseases necessitates the development of sensitive detection technologies that are able to elucidate the identity, potential pathogenicity, susceptibility, and viability of contaminating bacteria in a timely manner. For this purpose, we have designed a single multiplex PCR assay to simultaneously amplify 95 diagnostic regions (encompassing species/serogroup-specific, antimicrobial resistance, and known toxin markers) and combined it with a long oligonucleotide microarray to create a platform capable of rapidly detecting and discriminating the major human pathogenic species from the genus Vibrio: V. cholerae, V. parahaemolyticus, V. vulnificus, and V. mimicus. We were able to validate this strategy by testing 100 geographically and temporally distributed isolates and observed an excellent concordance between species- and serotype-level microarray-based identification and traditional typing methods. In addition to accurate identification, the microarray simultaneously provided evidence of antibiotic resistance genes and mobile genetic elements, such as sulfamethoxazole-trimethoprim constins and class I integrons, and common toxin (ctxAB, rtxA, hap, hlyA, tl, tdh, trh, vvhA, vlly, and vmhA) and pathogenicity (tcpA, type III secretion system) genes that are associated with pathogenic Vibrio. The versatility of this method was further underscored by its ability to detect the expression of known toxin and virulence genes from potentially harmful viable but nonculturable organisms. The results suggest that this molecular identification method provides rapid and definitive information that would be of value in epidemiological, environmental, and health risk assessment surveillance.

Anti-Infective Agents↗
PubMed2006

Emergence of multidrug-resistant Salmonella enterica serotype Newport in Minnesota.

We report a concurrent increase in the number of isolates of Salmonella enterica serotype Newport and the rate of multidrug resistance in S. Newport isolates from animal and human populations in Minnesota. Antimicrobial susceptibility and pulsed-field gel electrophoresis analysis demonstrated heterogeneity of isolates and showed that 1 pulsed-field gel electrophoresis cluster contained most of the multidrug-resistant isolates with a resistance pattern and most class 1 integron isolates, implying the clonal origin of the isolates.

Animals↗
PubMed2004

Antimicrobial resistance and serotype prevalence of Salmonella isolated from dairy cattle in the southwestern United States.

Mature dairy cattle were sampled over a 2-year period (2001-2002) on six farms in New Mexico and Texas. Fecal samples (n = 1560) were collected via rectal palpation and cultured for Salmonella, and one isolate from each positive sample was serotyped. Three isolates of each serotype, with the exception of Salmonella Newport (n = 12), were examined for susceptibility to 17 antimicrobial agents. Twenty-two different serotypes were identified from a total of 393 Salmonella isolates. Montevideo was the predominant serotype (27%) followed by Mbandaka (15%), Senftenberg (11.4%), Newport (6.4%), Anatum (4.8%), and Give (4.8%). Salmonella Typhimurium and Dublin, two frequently reported serotypes, accounted for only 1% of the observed serotypes in this study. Sixty-four percent of the serotypes were susceptible to all 17 antimicrobials, 14% were resistant to a single agent, and 22% were multiresistant (2-11 types of resistance). All isolates tested were susceptible to amikacin, apramycin, imipenem, ceftriaxone, nalidixic acid, and ciprofloxacin. The most frequent types of resistance were to sulfamethoxazole, tetracycline, streptomycin, kanamycin, chloramphenicol, and ampicillin (ranging from 8.9 to 22.4%). Serotypes demonstrating multiple resistance included Dublin and Give (resistant to three or more antibiotics), Typhimurium (resistant to five antibiotics), and Newport (four and two isolates resistant to six and nine antibiotics, respectively). Class 1 integrons were present in only two Salmonella Dublin isolates and one Salmonella Newport isolate. The most prevalent resistance patterns observed in this study were toward antimicrobial agents commonly used in cattle, while all Salmonella isolates were susceptible to ceftriaxone and ciprofloxacin, antibiotics used in human medicine.

Animals↗
PubMed2005

Nosocomial spread of Pseudomonas aeruginosa isolates expressing the metallo-beta-lactamase VIM-2 in a hematology unit of a French hospital.

Dissemination of metallo-beta-lactamases (carbapenemases) was investigated retrospectively among ceftazidimeand imipenem-resistant Pseudomonas aeruginosa isolates in a hematology unit in Marseilles, France, from September, 1995, to March, 1999. Sixteen clinical isolates and 23 environmental strains were identified, with a same bla (VIM-2) gene that encoded a carbapenemase identified in Southern Europe and South Korea. Five different genotypes were identified among clinical and environmental P. aeruginosa isolates all harboring an approximately 45-kb plasmid with bla (VIM-2)-positive class 1 integrons varying in structures. This study identified a hidden reservoir of carbapenemase producers.

Ceftazidime↗
PubMed2005

Comparison of multidrug resistance gene regions between two geographically unrelated Salmonella serotypes.

OBJECTIVES: The aim of this study was to identify chromosomally integrated genes conferring multidrug resistance to a Salmonella enterica (S.) serotype Typhimurium isolate, phage type DT193, isolated in Ireland and to compare them with resistance genes conferring plasmid-mediated multidrug resistance to a S. Enteritidis isolate from Italy. METHODS: A complete DNA sequence of the regions containing the resistance genes was obtained from the chromosome of the S. Typhimurium DT193 isolate and from the IncI plasmid of the S. Enteritidis isolate. The plasmid was also characterized by conjugation and incompatibility grouping. RESULTS: Two 10 kb multidrug resistance non-Salmonella Genomic Island 1 type clusters were independently identified in the S. Enteritidis plasmid and in the chromosome of the S. Typhimurium isolate. Detailed characterization identified an IP-type 2 integron containing a dfrA1-aadA1 gene cassette and other common resistance determinants derived from the RSF1010 plasmid. CONCLUSIONS: These multidrug resistance regions originate following chromosomal integration of key resistance markers encountered on plasmids circulating in other Salmonella serotypes. This mechanism of marker acquisition may have future implications for the evolution of similar structures in previously susceptible serotypes, leading to an increased public health risk.

Chromosome Mapping↗
PubMed2005

Emergence of plasmid-mediated resistance to quinolones in Enterobacteriaceae.

Although quinolone resistance results mostly from chromosomal mutations in Enterobacteriaceae, it may also be mediated by plasmid-encoded Qnr determinants. Qnr proteins protect DNA from quinolone binding and compromise the efficacy of quinolones such as nalidixic acid. Qnr proteins (QnrA-like, QnrB and QnrS) have been identified worldwide with a quite high prevalence among Asian isolates with a frequent association with clavulanic acid inhibited expanded-spectrum beta-lactamases and plasmid-mediated cephalosporinases. The qnrA genes are embedded in complex sul1-type integrons. A very recent identification of the origin of QnrA determinants in the water-borne species Shewanella algae underlines the role of the environment as a reservoir for this emerging threat. It may help to determine the location of in vivo transfer of qnrA genes. Further analysis of the role (if any) of quinolones for enhancing this gene transfer may be conducted. This could prevent the spread, if still possible, of this novel antibiotic resistance mechanism.

Amino Acid Sequence↗
PubMed2005

Genetic basis for dissemination of armA.

OBJECTIVES AND METHODS: armA is a novel plasmid-borne 16S rRNA methyltransferase that confers high-level resistance to 4,6-disubstituted deoxystreptamines. Recently, we have isolated from a high-level broad-spectrum aminoglycoside-resistant Escherichia coli animal isolate a plasmid, pMUR050, that bore the armA gene. In order to elucidate the genetic basis for the spread of armA, we have determined the complete nucleotide sequence of pMUR050. RESULTS: armA was borne by a complex transposon composite flanked by two direct repeats of IS26. The transposon composite included a class one integron with sul1 for resistance to sulphonamides and ant3''9 conferring resistance to spectinomycin-streptomycin, and a macrolide efflux pump and mefE/mel conferring high-level resistance to erythromycin. We identified in GenBank that another plasmid, pCTX-M3, from a Polish Citrobacter freundii human isolate, bore the same genetic structure, including armA. CONCLUSIONS: armA is present in human and animal isolates within a novel transposon composite. Further spread of armA between bacteria of diverse origin is to be expected.

Aminoglycosides↗
PubMed2005

Characterization of the highly variable region surrounding the bla(CTX-M-9) gene in non-related Escherichia coli from Barcelona.

OBJECTIVES: The dispersion of a clone, a plasmid or a mobile element carrying the bla(CTX-M-9) gene was evaluated in 30 Escherichia coli strains isolated in Barcelona between 1996 and 1999. The presence of the previously described orf513-bearing class 1 integron, In60, carrying the bla(CTX-M-9) gene, was also studied. METHODS: The clonality was analysed by pulsed-field gel electrophoresis. Plasmid analysis was performed by S1 digestion and hybridization with the CTX-M-9 probe. PCR mapping using specific designed primers was used to study the presence of In60 and In60-like structures. RESULTS: The clonality between the 30 strains was minor. The size of bla(CTX-M-9) carrying plasmids ranged between approximately 80 and 430 kb. One strain produced only a chromosome-encoded CTX-M-9 beta-lactamase. Thirty-six per cent of the strains showed differences with respect to the In60 structure due to an insertion or deletion events. CONCLUSIONS: These findings suggest that the bla(CTX-M-9) gene may be carried by a mobile element that disperses it between plasmids. The fast dispersion of the CTX-M-9 enzyme could therefore be due to both diffusion of plasmids and mobile elements.

Base Sequence↗
PubMed2005

Molecular epidemiology of emergent multidrug-resistant Salmonella enterica serotype Typhimurium strains carrying the virulence resistance plasmid pUO-StVR2.

OBJECTIVES: To evaluate the incidence of a distinct multidrug-resistant (MDR) grouping of Salmonella serotype Typhimurium strains carrying the hybrid virulence resistance plasmid pUO-StVR2, and its possible evolution in the region where it was first detected [Principality of Asturias (PA), Spain]. METHODS: pUO-StVR2-containing isolates were tentatively identified by two genetic markers: the bla(OXA-30) gene and the class 1 integron InH:2000 bp/bla(OXA-30)-aadA1a. Positive isolates were examined for resistance profile (RP), plasmid content, virulence profile (VP) and genomic polymorphisms using macrorestriction-PFGE. RESULTS: A total of 182 out of 248 Typhimurium clinical isolates recorded in the PA over 2001-02 were ampicillin-resistant and could be distributed into several MDR groupings. A MDR grouping carrying pUO-StVR2, with a defined RP (AMP/bla(OXA-30), CHL/catA1, [STR-SPT]/[strA/B,aadA1a], SUL/[sul1,sul2], TET/tet(B), qacEDelta1, merA, +/-TMP/dfrA12, and containing InH), was represented by 49 isolates. The VPs of these isolates (24 genes screened) differed from that of the type strain LT2 by the absence of the sopE1 and pef genes. Macrorestriction analysis established six combined XbaI/BlnI PFGE profiles, and supported a clonal relationship among most of the isolates. CONCLUSIONS: During 2001-02, the isolates carrying pUO-StVR2 constituted the second most frequent S. Typhimurium MDR grouping recorded in the PA, preceded only by the pandemic pentaresistant DT104. Polymorphisms on the genomic DNA, different phage types, different plasmid profiles and the detection of trimethoprim resistance in one isolate encoded by an additional plasmid, were consistent with both intra-cluster evolution and horizontal transfer of the hybrid plasmid.

Ampicillin Resistance↗
PubMed2002

Tn5041-like transposons: molecular diversity, evolutionary relationships and distribution of distinct variants in environmental bacteria.

A detailed study on the geographic distribution, molecular diversity and evolutionary relationships of 24 closely related variants of the Tn5041 transposon found among 182 mercury resistant environmental Gram-negative strains from the IMG-Hg Reference Collection is reported here. RFLP analysis, followed by the determination of partial DNA sequences, identified 14 distinct types of these transposons, which differed from each other by 1-7 single-event DNA polymorphisms. No polymorphisms were detected at the right arm of the transposons except an insertion of a new mobile DNA element carrying a mer operon (named the mer2 cassette) within the Tn5041 mer operon. According to the model presented here, the insertion occurred via homologous recombination with a circular form of the mer2 cassette. A total of 8 point mutations, 1 internal deletion, 2 end-involving deletions, 3 mosaic regions and 2 insertions were detected at the left arm of the transposons. The insertions were a transposon closely related to Tn21 but lacking the integron and a new group II intron (named INT5041C). Inspection of the geographic distribution of the Tn5041 variants suggested that at least three long-distance waves of dissemination of these variants had occurred, accompanied by homologous recombination between different Tn5041 lineages. Movements of circular DNAs by homologous recombination as a source of mosaic genes and new mer genes, and formation of unusual mosaics ending or beginning at the Tn5041 att site are discussed.

Bacteria↗
PubMed2005

Repetitive-DNA-element PCR fingerprinting and antibiotic resistance of pan-European multi-resistant Acinetobacter baumannii clone III strains.

In the present study, it was shown that repetitive-DNA-element PCR fingerprinting using the (GTG)5 primer [(GTG)5-PCR] is a rapid and reliable tool to genotypically differentiate members of the recently described pan-European multi-resistant Acinetobacter baumannii (MAB) clone III from the known MAB clones I and II. The identification of four new representatives of the MAB clone III dating from 1991 to 1993 by (GTG)5-PCR indicates that this clone has persisted in European hospitals since the beginning of the 1990s. Tetracycline (TET) resistance was found to be common among clone III strains, including one strain that also displayed resistance to minocycline. The TET-resistance phenotype in this MAB clone appeared to be strongly associated with the presence of the efflux-type gene tet(A), but the fact that some members lack this gene or have acquired an additional tet gene [i.e. tet(M)] suggests that the tet gene carriage in the pan-European clone III population may have diversified in time and space. In contrast, all clone III strains shared the previously described aminoglycoside resistotype encoded by the aminoglycoside-modifying genes aphA6 and the class 1 integron-associated aadB, which may point to the fact that these genes probably are more stably inherited in MAB clone III compared to tet genes.

Acinetobacter Infections↗
PubMed2006

Emergence and spread of two distinct clonal groups of multidrug-resistant Escherichia coli in a veterinary teaching hospital in Australia.

Multidrug-resistant Escherichia coli (MDREC) expressing AmpC beta-lactamases have emerged as a cause of opportunistic infections in dogs. Following a cluster of extraintestinal infections caused by two distinct clonal groups (CGs) of bla(CMY)-producing MDREC, a 12-month infection control study was undertaken at a veterinary teaching hospital in Brisbane, Australia. Swabs from the rectum of hospitalized dogs (n=780), hospital staff (n=16) and the hospital environment (n=220) were plated onto selective agar to obtain multidrug-resistant (MDR) coliforms. These were then tested by multiplex PCR for E. coli uspA, bla(CMY) and the class 1 integron-associated dfrA17-aadA5 gene cassette for rapid identification of MDREC CG 1 (positive for all three genes) and CG 2 (positive for uspA and bla(CMY) only). A total of 16.5 % of the dog rectal swabs and 4.1% of the hospital environmental swabs yielded MDREC, and on the basis of multiplex PCR, PFGE and plasmid profiling, these were confirmed to belong to either CG 1 or CG 2. Both CG 1 and CG 2 isolates were obtained from clinical cases of extraintestinal infection and rectal swabs from hospitalized dogs over the same period of time, whereas only CG 1 isolates were obtained from the hospital environment. Both CGs were prevalent during the first 6 months, but only CG 2 was isolated during the second 6 months of the study. Two isolates obtained from rectal swabs of staff working in the hospital belonged to CG 2, with one of the isolates possessing the same REDP as nine isolates from dogs, including six isolates associated with cases of extraintestinal infection. CG 1 isolates belonged to E. coli serotypes O162 : H-, OR : H- or Ont : H-, whereas CG 2 isolates belonged to O153 : HR, OR : HR or OR : H34. These results confirm that in this particular outbreak, canine MDREC were highly clonal and CG 2 MDREC may colonize both humans and dogs.

Animals↗
PubMed2006

CTX-M-producing Salmonella spp. in Hong Kong: an emerging problem.

Two Salmonella enterica serovar Typhimurium isolates and one S. enterica serovar Enteritidis isolate that were resistant to 4 microg cefotaxime ml-1 and produced CTX-M-type extended-spectrum beta-lactamases (ESBLs) were characterized from patients in Hong Kong during 2003-2004. The S. Typhimurium strain isolated in 2003 produced a CTX-M-9 ESBL and harboured a blaCTX-M-9 gene that was associated with a class I integron-containing gene cassette and orf513 similar to those of In60. The second S. Typhimurium strain and the S. Enteritidis strain, both isolated in 2004, produced CTX-M-14; the former also produced TEM-1. The blaCTX-M-14 gene in these two isolates was associated with the insertion sequence ISEcp1. The CTX-M genes were present on a transferable plasmid of 62, 70 or 92 kb. PFGE of XbaI-restricted total DNA from the two S. Typhimurium isolates indicated that they were not clonally related. These three isolates were also resistant to one of the other non-beta-lactam antimicrobial agents tested. This is the first report of a CTX-M-9 ESBL in Salmonella in Hong Kong and the presence of blaCTX-M-9 and blaCTX-M-14 in S. Typhimurium.

Adult↗