Molecular detection of ß-lactamase and integron genes in clinical strains of Klebsiella pneumoniae by multiplex polymerase chain reaction

Abstract INTRODUCTION: Infections caused by β-lactamase-producing gram-negative bacteria, such as Klebsiella pneumoniae, are increasing globally with high morbidity and mortality. The aim of the current study was to determine antimicrobial susceptibility patterns and the prevalence of antibiotic resistance genes (β-lactamase and integron genes) using multiplex PCR. METHODS One-hundred K. pneumoniae isolates were collected from different clinical samples. Antibiotic susceptibility testing was performed with thirteen different antibiotics. Multiplex-PCR was used to detect β-lactamase (bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC) and integron genes (int I, int II, and int III). RESULTS: The highest and lowest rate of resistance was exhibited against amikacin (93%) and imipenem (8%), respectively. The frequency of β-lactamase-positive K. pneumoniae was 37%, and the prevalence of the bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC genes was 38%, 24%, 19%, 12%, 6%, 11%, 33%, 0%, 28%, and 23%, respectively. Of the 100 isolates, eight (8%) were positive for class I integrons; however, class II and III integrons were not detected in any of the strains. CONCLUSIONS: These results indicate co-carriage of a number of β-lactamase genes and antibiotic resistance integrons on the same plasmids harboring multi-drug resistance genes. It seems that these properties help to decrease treatment complications due to resistant bacterial infections by rapid detection, infection-control programs and prevention of transmission of drug resistance.

A Novel Integron Gene Cassette Harboring VIM-38 Metallo-β-lactamase in a Clinical Pseudomonas aeruginosa Isolate

No abstract available.

The sul1 Gene in Stenotrophomonas maltophilia With High-Level Resistance to Trimethoprim/Sulfamethoxazole

Emerging resistance to trimethoprim/sulfamethoxazole (SXT) poses a serious threat to the treatment of Stenotrophomonas maltophilia infections. We determined the prevalence and molecular characteristics of acquired SXT resistance in recent clinical S. maltophilia isolates obtained from Korea. A total of 252 clinical isolates of S. maltophilia were collected from 10 university hospitals in Korea between 2009 and 2010. Antimicrobial susceptibility was determined by using the CLSI agar dilution method. The sul1, sul2, and sul3 genes, integrons, insertion sequence common region (ISCR) elements, and dfrA genes were detected using PCR. The presence of the sul1 gene and integrons was confirmed through sequence analysis. Among the 32 SXT-resistant isolates, sul1 was detected in 23 isolates (72%), all of which demonstrated high-level resistance (> or =64 mg/L) to SXT. The sul1 gene (varying in size and structure) was linked to class 1 integrons in 15 of the 23 isolates (65%) harboring this gene. None of the SXT-susceptible isolates or the SXT-resistant isolates with a minimum inhibitory concentration of 4 and 8 mg/L were positive for sul1. Moreover, the sul2, sul3, and dfrA genes or the ISCR elements were not detected. The sul1 gene may play an important role in the high-level SXT resistance observed in S. maltophilia.

Detection of integrons in gram positive and non fermentative gram negative isolates from intensive care unit of Zagazig university hospitals

Integrons are mobile genetic elements considered to play a role in transmission of antibiotic resistance genes with limited studies concerning this issue. were first to detect the prevalence of integrons in Gram positive and non fermentative Gram negative isolates from Intensive Care Unit of Zagazig University Hospitals and to investigate the association between integrons carriage and antimicrobial susceptibility of bacterial isolates. Culture, API identification and antibiotic susceptibility testing were done to all isolates. PCR using primers for integrons Class I, II and III followed by RFLP using RsaI enzyme was applied to all Gram-positive and non-fermentative Gram-negative isolates. Out of 183 clinical samples: 40 non fermentative Gram negative and 33 Gram positive bacteria were detected. The most predominant isolates of both groups were Pseudomonas aeruginosa [62.5%] and Staph aureus [69.7%] respectively. A high statistical significant difference regarding association between amikacin, spectinomycin and ciprofloxacin resistance, and integron existence was found in P.aeruginosa. Meanwhile, ceftazidime, aztreonam, cefotaxime, tetracycline, piperacillin and Sulphamethoxazole-trimethoprim resistance were high significantly associated with integron existance in isolated A. baumannii. A high statistically significant difference exists between integrons carriage and multidrug resistant pathogens

Investigation of class 1 integrons in Klebsiella pneumoniae clinical and microbiota isolates belonging to different phylogenetic groups in Recife, State of Pernambuco

Introduction The high prevalence of Klebsiella pneumoniae infections is related to the ability of K. pneumoniae to acquire and disseminate exogenous genes associated with mobile elements, such as R plasmids, transposons and integrons. This study investigated the presence of class 1 integrons in clinical and microbiota isolates of K. pneumoniae belonging to different phylogenetic groups and correlated these results with the antimicrobial resistance profiles of the studied isolates. Methods Of the 51 isolates of K. pneumoniae selected for this study, 29 were from multidrug-resistant clinical isolates, and 22 were from children's microbiota. The susceptibility profile was determined using the disk diffusion method, and class 1 integrons were detected through polymerase chain reaction (PCR). Results The results showed that none of the 22 microbiota isolates carried class 1 integrons. Among the 29 clinical isolates, 19 (65.5%) contained class 1 integrons, and resistance to sulfamethoxazole/trimethoprim was identified in 18 of these isolates (94.7%). Among the K. pneumoniae isolates with class 1 integrons, 47% belonged to the KpI phylogenetic group, and one isolate (14.3%) carrying these genetic elements belonged to the KpIII group. Conclusions The wide variety of detected class 1 integrons supports the presence of high rates of antimicrobial resistance, genetic variability, and rapid dissemination of beta-lactamase genes among K. pneumoniae clinical isolates in recent years in hospitals in Recife-PE, Brazil. The findings of this study indicate that the surveillance of K. pneumoniae integrons in clinical isolates could be useful for monitoring the spread of antibiotic resistance genes in the hospital environment. .

Epidemiological Characterizations of Class 1 Integrons from Multidrug-Resistant Acinetobacter Isolates in Daejeon, Korea

BACKGROUND: Multidrug-resistant (MDR) Acinetobacter spp. acquire antimicrobial agent-resistance genes via class 1 integrons. In this study, integrons were characterized to investigate the antimicrobial resistance mechanisms of MDR Acinetobacter isolates. In addition, the relationship between the integron type and integron-harboring bacterial species was analyzed by using epidemiological typing methods. METHODS: Fifty-six MDR Acinetobacter spp.-A. baumannii (N=30), A. bereziniae (N=4), A. nosocomialis (N=5), and A. pittii (N=17)-were isolated. The minimum inhibitory concentrations (MICs) were determined on the basis of the results of the Epsilometer test (Etest). PCR and DNA sequencing was performed to characterize the gene cassette arrays of class 1 integrons. Multilocus sequence typing (MLST) and repetitive extragenic palindromic sequence (REP)-PCR were performed for epidemiological typing. RESULTS: Class 1 integrons were detected in 50 (89.3%) of the 56 isolates, but no class 2 or 3 integron was found within the cohorts. The class 1 integrons were classified into 4 types: 2.3-kb type A (aacA4-catB8-aadA1), 3.0-kb type B (aacA4-blaI(MP-1)-bla(OXA-2)), 3.0-kb type C (bla(VIM-2)-aacA7-aadA1), and 1.8-kb type D (aac3-1-bla(OXA-2)-orfD). Type A was most prevalent and was detected only in A. baumannii isolates, except for one A. bereziniae isolate; however, type B was amplified in all Acinetobacter isolates except for A. baumannii isolates, regardless of clone and separation time of the bacteria. CONCLUSIONS: Although class 1 integron can be transferred horizontally between unrelated isolates belonging to different species, certain types of class 1 integrons tend to transfer horizontally and vertically among A. baumannii or non-baumannii Acinetobacter isolates.

Resistência a tetraciclinas em isolados clínicos e ambientais de Escherichia coli, Klebsiella pneumoniae e Aeromonas spp: identificação e mapeamento do ambiente genético de genes tet / Tetracycline resistance in clinical and environmental isolates of Escherichia coli, Klebsiella pneumoniae and Aeromonas spp. identification and mapping of tet genes genetic context

A resistência bacteriana a antibióticos é aceita como um dos maiores problemas de saúde pública. As tetraciclinas são antibióticos de amplo espectro, e após seu uso indiscriminado observou-se o surgimento de bactérias resistentes, levando médicos e veterinários a diminuírem seu uso. Objetivos. Verificar o perfil de sensibilidade a tetraciclinas em isolados clínicos e ambientais de Escherichia coli, Klebsiella pneumoniae e Aeromonas spp., bem como pesquisar os principais genes tet associados à resistência a esta classe de antibióticos e determinar a potencial forma de disseminação destes genes através da caracterização de seu ambiente genético. Material e Métodos. Os perfis de sensibilidade à tetraciclina (TET), doxiciclina (DOX), minociclina (MIN) e tigeciclina (TGC) de 572 isolados foram obtidos através das técnicas de Disco-Difusão e Concentração Inibitória Mínima. Os isolados não-sensíveis à tetraciclina foram submetidos a reações de PCR para pesquisa de grupos Inc, genes tet e para a caracterização de seu ambiente genético pela pesquisa das integrases de classes 1, 2, 3 e 4, e dos elementos genéticos móveis Tn1721, IS26, Tn10 e ISAS5. Perfis de similaridade genética dos isolados foram obtidos através das técnicas de ERIC-PCR e PFGE. Após análise destes resultados 33 cepas foram selecionadas para as técnicas de S1-PFGE e transformação. Resultados. A partir dos 572 isolados 18,5 por cento foram resistentes à TET, 13,5 por cento à DOX, 8 por cento à MIN...

The antibiotic resistance is accepted as one of the major problems for public health. Tetracyclines are broad spectrum antibiotics, and its indiscriminate use promoted the emergence of resistant bacteria, leading physicians and veterinarians to decrease its use. Objectives. Verify the susceptibility of clinical and environmental isolates of Escherichia coli, Klebsiella pneumoniae and Aeromonas spp. to tetracyclines, and also search for the main tet genes associated with resistance to these antibiotics and determine the potential mechanism of tet genes dissemination by characterizing their genetic context. Material and Methods. Disk-Diffusion and Minimum Inhibitory Concentration tests were carried out in 572 isolates using tetracycline (TET), doxycycline (DOX), minocycline (MIN) and tigecycline (TGC). PCR was carried out in TET non-susceptible isolates for the detection of Inc groups, tet genes and its genetic context determination through the search of classes 1, 2, 3, and 4 integrases, and Tn1721, Tn10, IS26 and ISAS5 mobile genetic elements. Genetic similarities patterns were determined by ERIC-PCR and PFGE techniques. After analyzing the results 33 strains were selected for the S1-PFGE and transformation experiments. Results. From 572 isolates, 18.5 per cent were TET-resistant, 13.5 per cent DOX-resistant, 8 per cent MIN-resistant and none resistant to TGC. Twenty-two per cent and 16.3 per cent of clinical and environmental isolates were TET-resistant...

Molecular Epidemiology of Integron-Associated Antimicrobial Gene Cassettes in the Clinical Isolates of Acinetobacter baumannii from Northern Taiwan

BACKGROUND: The aims of this study were to understand the molecular epidemiology of integron-associated gene cassettes in Acinetobacter baumannii across four hospitals in northern Taiwan and to clarify the relationship between the presence of integrons and antibiotic-resistant phenotypes. METHODS: Sixty-five A. baumannii isolates, collected from the patients of four regional hospitals in northern Taiwan in 2009, were tested for the presence of integrons and their associated gene cassettes. The susceptibility difference between integron-positive and integron-negative A. baumannii strains was analyzed. Antibiotic-resistant phenotypes among A. baumannii with different types of gene cassette array combinations were also compared. RESULTS: Around 72% of the A. baumannii isolates carried class 1 integrase genes. Despite this, only three gene cassette arrays were found in the integrons. Integron-positive strains were significantly more resistant to all the tested antibiotics than the integrase-negative strains. All the four types of A. baumannii with different gene cassette array combinations were multidrug-resistant in nature. Gene cassette array aacA4-catB8-aadA1 existed in all the integron-positive A. baumannii isolates. Repetitive-sequence-based PCR (rep-PCR) results revealed the prevalence of one major cluster of imipenem-resistant A. baumannii strains (84%) in the four regional hospitals. CONCLUSIONS: The presence of integrons with associated antimicrobial resistance gene cassettes can be used as a representative marker of multidrug resistance in A. baumannii. Some prevalent gene cassette arrays may exist among epidemiologically unrelated A. baumannii strains.

Detection of integrons among multi-drug resistant (MDR) Escherichia coli strains isolated from clinical specimens in northern west of Iran

Transference of resistance determinants by integrons is one of the important factors that can contribute to the increase in multi-resistant bacteria. We determined the prevalence and class of integrons among multi-drug resistant (MDR) Escherichia coli strains isolated from clinical specimens in Tabriz teaching hospitals. Firstly, susceptibility of 140 isolates to 13 antibiotics was determined using the disc diffusion method. Then, prevalence and class of integrons was detected in MDR strains by PCR-RFLP. One hundred five (75 percent) of total 140 isolates were uropathogenic Escherichia coli (UPEC). Other pathotypes included were: diarrheagenic Escherichia coli (13; 9.3 percent), sepsis-associated E. coli (5; 3.6 percent) and newborn meningitis-associated E. coli (2; 1.4 percent). Antibiotic resistance patterns were as follows: amoxicillin 99.3 percent, gentamicin 33.6 percent, tetracycline 72.8 percent, ceftazidime 46.4 percent, co-trimoxazole 75 percent, imipenem 1.4 percent, ciprofloxacin 47.6 percent, norfloxacin 50.7 percent, cephalothin 77.8 percent, amikacin 12.1 percent, nitrofurantoin 12.9 percent, chloramphenicol 20.7 percent and nalidixic acid 60.7 percent. One hundred eighteen (84.2 percent) of tested isolates were multi-drug resistant. Prevalence of integrons was confirmed in 27.1 percent of MDR isolates. intI1 and intI2 were detected respectively in 22.05 percent and 5.08 percent of MDR strains. No intI3 was detected. Resistance to gentamicin, amikacin and chloramphenicol was significantly associated with the presence of integrons. These results showed high resistance of E. coli to routine antibiotics, however, in consideration of low prevalence of integrons among these strains, we can conclude that antibiotic resistance genes in these strains presumably carried on elements other than integrons.

Detección del gen codificante de la metalo-ß-lactamasa VIM-2 en un integrón de clase 1 asociado con el gen blaCTX-M-2 en un aislamiento clínico de Pseudomonas aeruginosa en el Uruguay: primera comunicación

Con el fin de analizar la presencia de metalo-ß-lactamasas en nuestro medio, se incluyeron en este estudio aislamientos de Pseudomonas aeruginosa causantes de infecciones nosocomiales en un centro hospitalario del Uruguay, en el período comprendido entre abril y setiembre de 2008. En un aislamiento se detectó la presencia del gen codificante de la metalo-ß-lactamasa VIM-2 asociado a un integrón de clase 1 y del gen codificante de una ß-lactamasa de espectro extendido CTX-M-2. Esta es la primera comunicación de la presencia de los genes blaCTX-M-2 y blaVIM-2 en un mismo aislamiento de P. aeruginosa. A pesar de que las carbapenemasas ya han sido ampliamente documentadas en varias partes del mundo, esta es la primera comunicación de una metalo-ß-lactamasa adquirida con actividad carbapenemasa en bacterias patógenas encontradas en el Uruguay.

VIM-2 metallo-ß-lactamase gen detection in a class 1 integron associated to blaCTX-M-2 in a Pseudomonas aeruginosa clinical isolate in Uruguay: first communication. In order to analyze the presence of metallo-ß-lactamase in our country, we included in this study Pseudomonas aeruginosa isolates causing nosocomial infections in a hospital from Uruguay. The presence of a metallo-ß-lactamase VIM-2 in a class 1 integron and of an extended spectrum -lactamase CTX-M-2 was detected in one isolate. This is the first report of both genes, blaCTX-M-2 and blaVIM-2,in the same P. aeruginosa isolate. Although carbapenemases have been extensively documented in the world, this is the first report of an acquired metallo-ß-lactamase with carbapenemase activity in pathogenic bacteria in Uruguay.

QRDR mutations, efflux system & antimicrobial resistance genes in enterotoxigenic Escherichia coli isolated from an outbreak of diarrhoea in Ahmedabad, India.

Background & objectives: Diverse mechanisms have been identified in enteric bacteria for their adaptation and survival against multiple classes of antimicrobial agents. Resistance of bacteria to the most effective fluoroquinolones have increasingly been reported in many countries. We have identified that most of the enterotoxigenic Escherichia coli (ETEC) were resistant to several antimicrobials in a diarrhoea outbreak at Ahmedabad during 2000. The present study was done to identify several genes responsible for antimicrobial resistance and mobile genetic elements in the ETEC strains. Methods: Seventeen ETEC strains isolated from diarrhoeal patients were included in this study. The antimicrobial resistance was confirmed by conventional disc diffusion method. PCR and DNA sequencing were performed for the identification of mutation in the quinolone resistance-determining regions (QRDRs). Efflux pump was tested by inhibiting the proton-motive force. DNA hybridization assay was made for the detection of integrase genes and the resistance gene cassettes were identified by direct sequencing of the PCR amplicons. Results: Majority of the ETEC had GyrA mutations at codons 83 and 87 and in ParC at codon 80. Six strains had an additional mutation in ParC at codon 108 and two had at position 84. Plasmid-borne qnr gene alleles that encode quinolone resistance were not detected but the newly described aac(6’)-Ib-cr gene encoding a fluoroquinolne-modifying enzyme was detected in 64.7 per cent of the ETEC. Class 1 (intI1) and class 2 (intI2) integrons were detected in six (35.3%) and three (17.6%) strains, respectively. Four strains (23.5%) had both the classes of integrons. Sequence analysis revealed presence of dfrA17, aadA1, aadA5 in class 1, and dfrA1, sat1, aadA1 in class 2 integrons. In addition, the other resistance genes such as tet gene alleles (94.1%), catAI (70.6%), strA (58.8%), blaTEM-1(35.2%), and aphA1-Ia (29.4%) were detected in most of the strains. Interpretation & conclusions: Innate gene mutations and acquisition of multidrug resistance genes through mobile genetic elements might have contributed to the emergence of multidrug resistance (MDR) in ETEC. This study reinforces the necessity of utilizing molecular techniques in the epidemiological studies to understand the nature of resistance responsible for antimicrobial resistance in different species of pathogenic bacteria.

Antimicrobial Resistance Determinants in Imipenem-nonsusceptible Acinetobacter calcoaceticus-baumannii Complex Isolated in Daejeon, Korea / 대한진단검사의학회지

BACKGROUND: Members of the Acinetobacter calcoaceticus-baumannii (Acb) complex are important opportunistic bacterial pathogens and present significant therapeutic challenges in the treatment of nosocomial infections. In the present study, we investigated the integrons and various genes involved in resistance to carbapenems, aminoglycosides, and fluoroquinolones in 56 imipenem-nonsusceptible Acb complex isolates. METHODS: This study included 44 imipenem-nonsusceptible A. baumannii, 10 Acinetobacter genomic species 3, and 2 Acinetobacter genomic species 13TU strains isolated in Daejeon, Korea. The minimum inhibitory concentrations (MICs) were determined by Etest. PCR and DNA sequencing were used to identify the genes that potentially contribute to each resistance phenotype. RESULTS: All A. baumannii isolates harbored the blaOXA-51-like gene, and 21 isolates (47.7%) co-produced OXA-23. However, isolates of Acinetobacter genomic species 3 and 13TU only contained blaIMP-1 or blaVIM-2. Most Acb complex isolates (94.6%) harbored class 1 integrons, armA, and/or aminoglycoside-modifying enzymes (AMEs). Of particular note was the fact that armA and aph(3')-Ia were only detected in A. baumannii isolates, which were highly resistant to amikacin (MIC50> or =256) and gentamicin (MIC50> or =1,024). In all 44 A. baumannii isolates, resistance to fluoroquinolones was conferred by sense mutations in the gyrA and parC. However, sense mutations in parC were not found in Acinetobacter genomic species 3 or 13TU isolates. CONCLUSIONS: Several differences in carbapenem, aminoglycoside, and fluoroquinolone resistance gene content were detected among Acb complex isolates. However, most Acb complex isolates (87.5%) possessed integrons, carbapenemases, AMEs, and mutations in gyrA. The co-occurrence of several resistance determinants may present a significant threat.

blaCTX-M-2 and blaCTX-M-28 extended-spectrum â-lactamase genes and class 1 integrons in clinical isolates of Klebsiella pneumoniae from Brazil

Twenty-eight Klebsiella pneumoniae clinical isolates that exhibited an extended-spectrum cephalosporin-resistance profile from a city in the Northeast of Brazil were analysed by PCR and DNA sequencing in order to determine the occurrence of blaCTX-M genes and class 1 integrons. We determined the occurrence of the blaCTX-M-2 gene in six K. pneumoniae isolates and describe the first detection of the blaCTX-M-28 gene in South America. Seven isolates carried class 1 integrons. Partial sequencing analysis of the 5'-3'CS variable region in the class 1 integrons of three isolates revealed the presence of aadA1, blaOXA-2 and dfr22 gene cassettes.

Occurrence and composition of class 1 and class 2 integrons in clinical and environmental O1 and non-O1/non-O139 Vibrio cholerae strains from the Brazilian Amazon

This study identified and characterised class 1 and 2 integrons in clinical and environmental Vibrio cholerae O1 and non-O1/non-O139 strains isolated from the Brazilian Amazon. The aadA2 and aadA7 gene cassettes were found in class 1 integrons in two genotypes of environmental V. cholerae non-O1/non-O139. Empty integrons were found in strains from the Brazilian cholera epidemic. A class 2 integron was detected in one strain from the V. cholerae Amazonia lineage harbouring sat1 and aadA1 genes. All isolates were resistant to aminoglycosides, indicating aadA functionality. These findings suggest that environmental bacteria act as cassette reservoirs that favour the emergence of resistant pathogens.

Caracterización de la región variable de integrones clase 1 presentes en cepas nosocomiales de Klebsiella pneumoniae / Characterization of the variable region within class 1 integrons in Klebsiella pneumoniae nosocomials strains

Background: Integrons are responsible for the capture and dissemination of resistance genes in Gram-negative bacteria. Aim: To characterize the variable region within class 1 integrons in nosocomial strains of Klebsiella pneumoniae. Material and Methods: Twenty-nine Klebsiella pneumoniae strains isolated from hospitalized patients were analyzed. The variable region of class 1 integrons was characterized by polymerase chain reaction and direct sequencing. Genetic localization of class 1 integrons was determined by bacterial conjugation. Results: Ten strains contained class 1 integrons, with sizes ranging from 750 to 2000 base pairs. One integrant element was present in nine strains and two elements in one single strain. Integrons were associated to plasmids. Cassettes aadA, aac(6)-Iq, orfD, dfrA]7, aadA5 and aadB were found. Conclusions: The presence of class 1 integrons may play an important role in the dissemination of hospital resistance against amino glycosides.

Genetic Basis of Multidrug-resistant Acinetobacter baumannii Clinical Isolates from Three University Hospitals in Chungcheong Province, Korea / 대한진단검사의학회지

BACKGROUND: The emergence of multidrug-resistant (MDR) Acinetobacter baumannii as an important opportunistic pathogen has given rise to significant therapeutic challenges in the treatment of nosocomial infections. In the present study, we assess the antibiotic resistance mechanisms of MDR A. baumannii strains by estimating the prevalence of antibiotic resistance determinants, including integrons, beta-lactamases, str genes, and gyrA and parC mutations. METHODS: Thirty-five MDR A. baumannii clinical isolates were collected from 3 Korean university hospitals over a 2-yr period. A. baumannii was confirmed by rpoB gene analysis. For each isolate, the minimal inhibitory concentrations (MICs) of 9 antibiotics were determined by the agar dilution method. PCR and DNA sequencing were used to identify the genes that potentially contribute to each resistance phenotype. RESULTS: Of the 35 MDR A. baumannii isolates examined, 7 antibiotic resistance gene determinants were detected. These resistance gene determinants included the gene bla(OXA-23), with an upstream element ISAba1 to promote increased gene expression and subsequent resistance to carbapenems, in 8 isolates (22.9%); aacA4, located within class 1 integrons, in 7 isolates (20.0%); and fluoroquinolone resistance conferred by gyrA and parC sense mutations in 31 isolates. CONCLUSIONS: Of the 35 MDR A. baumannii isolates, 26 (74.3%) from both outbreak and sporadic cases possessed at least 4 of the 7 antibiotic resistance gene determinants that give rise to the MDR phenotype. The co-occurrence of several resistance determinants may present a significant threat.

Analysis of Acquired Resistance Genes in Stenotrophomonas maltophilia / 대한진단검사의학회지

BACKGROUND: Stenotrophomonas maltophilia is a gram-negative bacillus and a nosocomial pathogen in immunocompromised patients. Trimethoprim/sulfamethoxazole (TMP/SMX) is the drug of choice for treating S. maltophilia infection; however, resistance to TMP/SMX is increasing. In this study, we investigated the relationship between the incidence of TMP/SMX resistance and the presence of sul genes and mobile elements. METHODS: A total of 120 S. maltophilia isolates were collected from 3 university hospitals between April 2007 and April 2009. Antimicrobial susceptibilities were determined using the disk diffusion method. PCR and DNA sequencing were conducted for the detection of sul1, sul2, class 1 integron, and ISCR2 element. Repetitive extragenic palindromic sequence-based PCR (REP-PCR) was carried out to evaluate the genetic relatedness. RESULTS: The TMP/SMX-resistant (R) isolates harbored a significantly higher proportion of sul1 gene and class 1 integron than TMP/SMX-susceptible (S) isolates (P<0.001). Seventeen of 28 isolates with sul1 also had a class 1 integron, but none of the isolates without sul1 had a class 1 integron. The identified gene cassettes within class 1 integrons include aacA4, aadA1, aac6'-II, and qac. None of the 120 isolates carried sul2, glmM, or ISCR2 element. REP-PCR did not show any genetic relatedness among the isolates. CONCLUSIONS: In Korea, the resistance of S. maltophilia isolates to TMP/SMX is due to sul1 within a class 1 integron rather than to sul2. The class 1 integron also harbors multiple antibiotic resistance genes in addition to sul1, and therefore it could mediate multidrug resistance in S. maltophilia.

Serovariedades de Salmonella enterica subespecie enterica en porcinos de faena y su resistencia a los antimicrobianos / Serovars of Salmonella enterica subspecies enterica and its antimicrobial resistance in slaughterhouse pigs

Se realizó un estudio para determinar la prevalencia de Salmonella y sus serovariedades en cerdos de faena, para evaluar sus perfiles de resistencia a los antimicrobianos y para conocer la presencia de integrones de clase 1 como posibles reservorios de resistencia. A partir de un total de 386 muestras de porcinos provenientes de cuatro frigoríficos de las provincias de Buenos Aires y de Santa Fe (Argentina), se identificaron 93 (24,1%) cepas de Salmonella enterica subespecie enterica, 52 (55,9%) de contenido cecal y 41 (44,1%) de nódulo linfático ileocecal. Se hallaron 13 serovariedades de S. enterica, las más prevalentes fueron S. Schwarzengrund, S. Heidelberg, S. subespecie I 6,8:e,h:-, S. Derby y S. Bredeney. Se probaron 15 antimicrobianos por el método de dilución en agar: amikacina, gentamicina, ciprofloxacina, cefalotina, cefotaxima, enrofloxacina, fosfomicina, polimixina-B, tetraciclina, cloranfenicol, estreptomicina, trimetoprima-sulfametoxazol, ampicilina, nitrofurantoína y ácido nalidíxico. Según se estableció mediante la determinación de la CIM, el 73% de las cepas de S. enterica subespecie enterica fueron sensibles a todos los antimicrobianos probados. Se observó resistencia a tetraciclina en 24 (25,8%) de las 93 cepas, a cloranfenicol en 22 (23,7%), a estreptomicina en 22 (23,7%) a trimetoprima-sulfametoxazol en 20 (21,5%), a ampicilina en 18 (19,4%), a nitrofurantoína en 3 (3,2%) y a ácido nalidíxico en 3 (3,2%). Algunos aislamientos de S. Typhimurium, S. Heildelberg, S. Derby y S. Orion presentaron multirresistencia y portaban el gen de la integrasa clase 1. Los mayores porcentajes de resistencia correspondieron a los antimicrobianos habitualmente utilizados en veterinaria y en las explotaciones porcinas.

A study was carried out in order to determine the prevalence of Salmonella and its serovars among porcine slaughterhouses, to evaluate the antimicrobial resistance profiles and to know the presence of class 1 integrons as possible reservoir of resistance. From a total of 386 samples from four porcine slaughterhouses of Buenos Aires and Santa Fe Provinces (Argentina), 93 (24,1%) Salmonella enterica subspecies enterica strains were identified, 52 (55,9%) from cecal contents and 41 (44,1%) from ileocecal lymph nodes. Thirteen serovars of S. enterica were found, the most prevalent were: S. Schwarzengrund, S. Heidelberg, S. subspecie I 6,8:e,h:-, S. Derby and S. Bredeney. Fifteen antimicrobials by the agar dilution method were tested: amikacin, gentamicin, ciprofloxacin, cephalotin, cefotaxime, enrofloxacin, fosfomycin, polimixin-B, tetracycline, chloramphenicol, streptomycin, trimethoprim-sulfamethoxazole, ampicillin, nitrofurantoin, and nalidixic acid. According to the CIM determination, 73% Salmonella enterica subspecies enterica strains were sensible to all the antimicrobials tested. Antimicrobial resistance was observed to tetracycline in 24 (25,8%) of 93 strains, to chloramphenicol in 22 (23,7%), to streptomycin in 22 (23,7%), to trimethoprim-sulfamethoxazole in 20 (21,5%), to ampicillin in 18 (19,4%), to nitrofurantoin in 3 (3,2%) and to nalidixic acid in 3 (3,2%). Some isolates of S. Typhimurium, S. Heidelberg, S. Derby, S. Orion showed multidrug resistance and carried the class 1 integrase gene. The highest percentage of resistance corresponded to the antimicrobials currently used in veterinary and porcine farms.

Characterization of class 1 integrons with unusual 3' conserved region from Salmonella enterica isolates.

The unusual 3' conserved sequence region of class 1 integrons was characterized in seven Salmonella isolates from swine and poultry. Three types of gene cassette arrays, aadA2-cmlA1-aadA1, sat-psp-aadA2-cm1A1-aadA1 and drfA12-orf-aadA2-cmlA1-aadA1, were found to be linked to a genetic organization qacH-IS440-sul3. All class 1 integrons were located on a conjugative plasmid that could be transferred to Escherichia coli. The results support the notion that the use of an antibiotic can select for resistance not only to that specific agent, but also to other unrelated antimicrobials including those that are no longer approved for use in food animal production.

Class 1 integrons and multidrug resistance among Escherichia coli isolates from human stools.

Three hundred and eighteen Escherichia coli isolates from stools of healthy volunteers and outpatients from a major university hospital in southern Thailand were tested for the presence of class 1 integrons using multiplex-PCR and for their susceptibility against 12 antimicrobial agents using standard disc diffusion method. Based on the presence of intl1, 162 isolates harbored class 1 integrons, which were more prevalent in isolates from outpatients compared with those from healthy volunteers. The majority (85%) of the isolates were resistant to at least one antimicrobial agent with the following percent resistance: streptomycin 66%, tetracycline 60%, sulphamethoxazole 59%, ampicillin 52%, trimethoprim/sulphamethoxazole 47%, kanamycin 30%, nalidixic acid 27%, ciprofloxacin 23%, norfloxacin 22%, amoxicillin/ clavulanic acid 16%, gentamicin 8%, and amikacin 2%. The most frequent pattern of multiresistant strains (11%) was sulphamethoxazole- trimethoprim/sulphamethoxazole -ampicillin-tetracycline-streptomycin. Multiple drug resistance was more frequent in integron-positive isolates (89%) than those in integron-negative E. coli (57%). These data indicate that human fecal E. coli is a reservoir of antibiotic-resistant genes that poses a significant risk of the spread of microbial resistance in the community.