Antimicrobial resistance patterns and integron carriage of Escherichia coli isolates causing community-acquired infections in Turkey

We aimed to observe antimiCrobial resistance patterns and integron carriage of Escherichia coli isolates causing community-acquired infections. Two hundred sixty-eight E. coli strains were obtained from outpatients with various infections at different polyclinics at the 82nd Year of State Hospital in Rize, Turkey. Susceptibility to antimicrobials was tested using a disk diffusion method. The presence of integrons was examined using PCR with specific primers. Positive PCR results were confirmed by sequencing. A broth mating method was used for conjugation assays. Extragenic palindromic-FOR was performed using the oligonucleotide primer BOXA1R. Resistance frequency for ampicillin,trimethoprim/sulfamethoxazole, and tetracycline was determined as 50.6%, 33.5%, and 36.8% respectively. No strains were resistant to amikacin. Seventy isolates were positive for the intll gene, of which 49 carried gene cassettes. Eleven isolates were positive for the int12 gene, eight of swhich carried gene cassettes. Seven gene cassettes (dfrAl, dfrA5, dfrA7, dfrA17, aadAl, aadA5, and sat2) were predominantly harbored in integrons. We detected conjugative plasmids harboring integrons in two E. coli strains. Four strain clusters were yielded by BOX-FOR fingerprints showing that they were clonally related. No apparent relationship occurred among class 1 and 2 integron-carrying strains. We conclude that integrons are widespread in genetically variable E. coli strains and will continue to mediate dissemination of resistance genes in the community.We aimed to observe antimiCrobial resistance patterns and integron carriage of Escherichia coli isolates causing community-acquired infections. Two hundred sixty-eight E. coli strains were obtained from outpatients with various infections at different polyclinics at the 82nd Year of State Hospital in Rize, Turkey. Susceptibility to antimicrobials was tested using a disk diffusion method. The presence of integrons was examined using PCR with specific primers. Positive PCR results were confirmed by sequencing. A broth mating method was used for conjugation assays. Extragenic palindromic-FOR was performed using the oligonucleotide primer BOXA1R. Resistance frequency for ampicillin, trimethoprim/sulfamethoxazole, and tetracycline was determined as 50.6%, 33.5%, and 36.8% respectively. No strains were resistant to amikacin. Seventy isolates were positive for the intll gene, of which 49 carried gene cassettes. Eleven isolates were positive for the int12 gene, eight of swhich carried gene cassettes. Seven gene cassettes (dfrAl, dfrA5, dfrA7, dfrA17, aadAl, aadA5, and sat2) were predominantly harbored in integrons. We detected conjugative plasmids harboring integrons in two E. coli strains. Four strain clusters were yielded by BOX-FOR fingerprints showing that they were clonally related. No apparent relationship occurred among class 1 and 2 integron-carrying strains. We conclude that integrons are widespread in genetically variable E. coli strains and will continue to mediate dissemination of resistance genes in the community.This work was supported by Recep Tayyip Erdogan University Research Fund grants BAP-2012.106.01.11 and BAP-2011. 102.03.3

Antibiotic resistance profiles of enteric cacteria isolated from Kucukcekmece Lagoon (Istanbul-Turkey)

Sivri, Nuket/0000-0002-4269-5950; SANDALLI, Cemal/0000-0002-1298-3687WOS: 000314625500019The aim of this study is to find out the density of the fecal bacteria and to analyze resistance to antimicrobials of Gram negative bacilli isolated from the Kucukcekmece Lagoon, Istanbul. Samples were taken monthly from June 2006 to June 2008 and a total of 232 Gram negative bacilli were isolated. Chloramphenicol, tetracycline, nalidixic acid, ampicillin, imipenem, ceftazidime, amikacin, streptomycin and amoxicillin + clavulanic acid were used in antimicrobial susceptibility tests. Susceptibility to trimethoprim + sulfamethoxazole was also examined in only integron-bearing organisms. the antibiotic resistance tests resulted in bacteria being the most resistant against ampicillin (76.29%) and the most sensitive against amikacin (93.56%). of 232 isolates, 20 (8.6%) coliforms harbored class 1 and/or class 2 integrons. DNA sequencing showed that variable regions of the integrons harbored various gene cassettes; dfrA12, dfrA15, dfrA17, aadA1, aadA2, aadA5, b/a(OXA-30) and sat2. Integrons were found in bacteria from all sampling areas except 12 and D3. in this study, the determination of bacterial identification of the species of Gram negative bacilli and their Antibiotic Resistance Profiles in the Kucukcekmece Lagoon for the first time was investigated. A finding indicates that there is a heavy fecal pollution in this lagoon environment, which might probably be resulted from the intensive anthropogenic facilities. the risk to public health could be the transfer of antibiotic resistance determinants from the bacterial isolates to normal microbiota bacteria of humans unless the effective precautions such as water treatment plants are taken.Research Fund of the University of IstanbulIstanbul University [BAP-543/05052006]; TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [105Y116]This work was supported by the Research Fund of the University of Istanbul (Project Number : BAP-543/05052006) and the TUBITAK (Project Number : 105Y116)

Molecular epidemiological analysis of integron gene cassettes and tetA/tetB/tetD gene associations in Escherichia coli strains producing extended-spectrum beta-lactamase (ESBL) in urine cultures

Background. Epidemiological studies of tetracycline (TE) resistance genes and integron gene cassettes, particularly in urine samples, are limited in Turkey. Objectives. To investigate antibiotic susceptibility profiles, extended-spectrum beta-lactamase (ESBL) positivity, tet gene types, class-I/-II integron gene cassettes, and clonal relationships among tet-resistant isolates of Escherichia coli from urine cultures of outpatients. Materials and methods. Isolates were identified using conventional methods and the automated Vitek (R) 2 Compact system. Antimicrobial susceptibility was performed for 19 antibiotics. The ESBL production was performed using the Kirby-Bauer disk diffusion test. The double disk synergy test was used for confirmatory testing. Polymerase chain reaction (PCR) was used to determine the presence of class-I/-II integron gene cassettes and tetA, tetB and tetD resistance genes. The pulsed-field gel electrophoresis typing was performed to identify clonal relations. Results. A total of 121 isolates were obtained and found to be resistant or sensitive to ampicillin and amikacin/imipenem. Resistance to ceftazidime, cefotaxime and ceftriaxone was determined to be 31.3%, 77.6% and 83.1%, respectively. Tetracycline resistance was detected in 82 isolates, mostly caused by the tetB gene. No tet gene was detected in the remaining 39 isolates. Although 64 out of 82 isolates carried a class-I integron, only 4 had a class-II integron (with sizes of 800-2900 base pairs). Furthermore, tet genes were identified with different size class-I integron gene cassettes. However, tet genes were not detected in any isolate identified with integron gene cassette II. Clonally, the isolates were found to be related in subgroups because they were community-acquired. Conclusions. This study showed thatthe tetB gene is most commonly found in E. coli isolates grown in urine samples from the Turkish population

Enterrobacteriaceae'nin klinik izolatları arasındaki temel ve SHV tipi Beta-Laktamaz genler taşıyan direnç plasmidlerinin horizontol yaygınlaştırılması

Genişletilmiş spektrumlu beta-laktamaz (ESBL) üreten bakteriler, dünya çapında artan sıklıkta izole edilmiştir. ESBL'nin ifadesi genellikle çoklu ilaç direnci ve direnç plazmidleri tarafından yayılma ile ilişkilidir. 2000 yılında iki aylık bir süre boyunca, enterobakteriyel suşların 133 klinik izolatı, Türkiye'deki bir üniversite hastanesinde ayaktan ve yatan hastalardan rastgele toplandı. ESBL üreten suşlar, çift disk sinerji (DDS) testi ile belirlendi. Escherichia coli (n = 9), Klebsiella pneumoniae (n = 7), Klebsiella oxytoca (n = 2) ve Enterobacter aerogenes (n = 2) dahil olmak üzere 20 ESBL üreten suş (% 15) tespit edildi ve direnç aktarım özellikleri açısından daha fazla analiz edildi direnç genlerinin plazmid profili ve doğası. Plazmid transfer deneyleri, broth çiftleşme teknikleri kullanılarak yapıldı. TEM- ve SHV-genleri, polimeraz zincir reaksiyonu (PCR) ve spesifik problar kullanılarak hibridizasyon ile analiz edildi. Salgın plazmidlerin saptanmasında R plazmidlerinin EcoRI restriksiyon enzim analizleri kullanıldı. EcoRI restriksiyon enzim analizi ile on dört plazmid profili (A, B1, B2, C1 ve C2 ila L) elde edildi. Bu plazmitlerin çoğunun, PCR ile hem TEM hem de SHV'den türetilmiş genleri taşıdığı tespit edildi ve her bir genin hibridizasyon deneyi ile lokalize edilmesiyle doğrulandı. Epidemiyolojik kanıtlar, bu hastanede çoklu ilaca dirençli enterobakteriyel cinsler ve türler arasında konjugatif R plazmidlerinin görünür bir yatay yayılımı olduğunu gösterdi. EcoRI restriksiyon enzim analizi ile on dört plazmid profili (A, B1, B2, C1 ve C2 ila L) elde edildi. Bu plazmitlerin çoğunun, PCR ile hem TEM hem de SHV'den türetilmiş genleri taşıdığı tespit edildi ve her bir genin hibridizasyon deneyi ile lokalize edilmesiyle doğrulandı. Epidemiyolojik kanıtlar, bu hastanede çoklu ilaca dirençli enterobakteriyel cinsler ve türler arasında konjugatif R plazmidlerinin belirgin bir yatay yayılımı olduğunu göstermiştir. EcoRI restriksiyon enzim analizi ile on dört plazmid profili (A, B1, B2, C1 ve C2 ila L) elde edildi. Bu plazmitlerin çoğunun, PCR ile hem TEM hem de SHV'den türetilmiş genleri taşıdığı tespit edildi ve her bir genin hibridizasyon deneyi ile lokalize edilmesiyle doğrulandı. Epidemiyolojik kanıtlar, bu hastanede çoklu ilaca dirençli enterobakteriyel cinsler ve türler arasında konjugatif R plazmidlerinin görünür bir yatay yayılımı olduğunu gösterdi.The extended-spectrum ß-lactamase (ESBL)-producing bacteria have been isolated at increasing frequency worldwide. Expression of ESBL is often associated with multidrug resistance and dissemination by resistance plasmids. During a two-month period in 2000, 133 clinical isolates of enterobacterial strains were randomly collected from outpatients and inpatients at a university hospital in Turkey. The ESBL producing strains were determined by double-disk synergy (DDS) testing. Twenty ESBL producing strains (15%) including Escherichia coli (n = 9), Klebsiella pneumoniae (n = 7), Klebsiella oxytoca (n = 2) and Enterobacter aerogenes (n = 2) were detected and further analyzed for their resistance transfer features, plasmid profile and nature of the resistance genes. Plasmid transfer assays were performed using broth mating techniques. TEM- and SHV- genes were analyzed by polymerase chain reaction (PCR) and hybridization using specific probes. EcoRI restriction enzyme analyses of R plasmids were used in the detection of epidemic plasmids. Fourteen plasmid profiles (A, B1, B2, C1, and C2 to L) were obtained with EcoRI restriction enzyme analysis. Most of these plasmids were detected to carry both TEM- and SHV-derived genes by PCR, and confirmed by localizing each gene by hybridization assay. Epidemiological evidence indicated that there was an apparent horizontal dissemination of conjugative R plasmids among multidrug-resistant enterobacterial genera and species in this hospital

Determination and molecular analysis of antibiotic resistance in Gram-negative enteric bacteria isolated from Pelophylax sp. in the Eastern Black Sea Region (vol 69, pg 223, 2021)

The sentence 'However, two of three quinolone-resistant Klebsiella strains showed the novel amino acid substitution in the gyrA gene resulting in Ser83Asp and Asp87Glu.' should be corrected to 'However, one of three quinolone-resistant Klebsiella strains showed the novel amino acid substitution in the gyrA gene resulting in Ser83Thr.