Antibiotic resistance profiles and integrons in Enterobacteriaceae from the riverside of Evros-Ardas with respect to chemical and waste pollution (original) (raw)

Incidence of class 1 integrons in multiple antibiotic-resistant Gram-negative copiotrophic bacteria from the River Torsa in India

Research in microbiology, 2006

The presence of class 1 integrons in multiple-antibiotic-resistant (MAR) Gram-negative copiotrophic bacteria from the River Torsa in India was detected using a polymerase chain reaction (PCR)-based screening method. Among 100 isolates that were resistant to at least five of the twelve antibiotics tested, 40 carried class 1 integrons, with inserted DNA regions of 0.7–3.2 kb. Carriage of integrons in strains of higher MAR index was found to be statistically significant. DNA sequencing was used to identify the genetic content of the integron-variable regions. In addition to the identification of gene cassettes dfrA1, dfrA5, dfrA7, dfrA17 and a variant of dfrA12 for trimethoprim, aac(6′)-Ib for amikacin and tobramycin and aadA1 and aadA6 for streptomycin and spectinomycin resistance, a novel ORF predicted from a sequence of Morganella sp. TR 90 bearing homology with the Vibrio cholerae dfrA1 gene cassette was characterized. To our knowledge, this is the first report of the incidence and abundance of class 1 integrons in copiotrophic river water bacteria from India.

Antibiotic resistance and prevalence of class 1 and 2 integrons in Escherichia coli isolated from two wastewater treatment plants, and their receiving waters (Gulf of Gdansk, Baltic Sea, Poland)

2014

In this study, antimicrobial-resistance patterns were analyzed in Escherichia coli isolates from raw (RW) and treated wastewater (TW) of two wastewater treatment plants (WWTPs), their marine outfalls (MOut), and mouth of the Vistula River (VR). Susceptibility of E. coli was tested against different classes of antibiotics. Isolates resistant to at least one antimicrobial agent were PCR tested for the presence of integrons. Ampicillin-resistant E. coli were the most frequent, followed by amoxicillin/clavulanate (up to 32 %), trimethoprim/sulfamethoxazole (up to 20 %), and fluoroquinolone (up to 15 %)-resistant isolates. Presence of class 1 and 2 integrons was detected among tested E. coli isolates with rate of 32.06 % (n = 84) and 3.05 % (n = 8), respectively. The presence of integrons was associated with increased frequency of resistance to fluoroquinolones, trimethoprim/sulfamethoxazole, amoxicillin/clavulanate, piperacillin/tazobactam, and presence of multidrug-resistance phenotype. Variable regions were detected in 48 class 1 and 5 class 2 integron-positive isolates. Nine different gene cassette arrays were confirmed among sequenced variable regions, with predominance of dfrA1-aadA1, dfrA17-aadA5, and aadA1 arrays. These findings illustrate the importance of WWTPs in spreading of resistance genes in the environment and the need for inclusion of at least monitoring efforts in the regular WWTP processes.

Occurrence of antibiotic resistance and class 1, 2 and 3 integrons in Escherichia coli isolated from a densely populated estuary (Seine, France)

FEMS Microbiology Ecology, 2000

Over 6 years, Escherichia coli were isolated from water samples from seven Seine estuary stations, characterized by a densely populated watershed (654 isolates). Resistances of these E. coli to 16 antibiotics were determined and compared with the same resistances in E. coli isolated from a small stream (120 isolates) and from the treated effluent of the largest estuary wastewater treatment plant (123 isolates). Between 30.2% and 56.6% of the estuary isolates were resistant, whatever the station or time of sampling; of these, 60.5-80% were resistant to at least two and up to 12 antibiotics. In the three contrasting sites, resistances to tetracycline, amoxicillin and ticarcillin were the commonest. DNA was extracted from 279 estuary isolates (January 2006) and class 1, 2 and 3 integrons were detected by multiplex real-time PCR and confirmed by classic PCR. IntI1 and intI2 genes were found in 11% of isolates. No intI3 gene was detected. The variable regions of the class 1 and 2 integrons sequenced contained predominantly gene cassettes aadA and dfr. However, for slightly over half of the E. coli isolates exhibiting the class 1 integron, the variable region could not be amplified, because part of the 3 0 conserved sequence was missing.

Antibiotic Resistance and intI1 Carriage in Waterborne Enterobacteriaceae

Water, Air, & Soil Pollution, 2016

The spread of antimicrobial resistance (AMR) is a global concern, high research priority being given to the environmental contamination, as the prevalence of organisms exhibiting AMR continues to increase. Multiresistant bacteria carrying different mobile genetic elements have been detected in sites with different degrees of urbanization, surface waters receiving insufficiently treated effluents being at high risk. The aim of the present study was to investigate the loads, antibiotic susceptibility, and class 1 integron carriage of Enterobacteriaceae isolated from surface waters and wastewaters around a large Romanian city. Searching for a valuable genetic marker of the displayed antibiotic resistance, the link between the AMR and the presence of int1I gene was explored in a total of 166 waterborne strains. Overall, amoxicillin-clavulanate resistance displayed the highest frequency (71.1 %), followed by ampicillin (63.9 %), cefuroxime (21.1 %), ciprofloxacin (17.5 %), cefotaxime (15.7 %), ceftriaxone (10.8 %), and gentamicin (6.6 %). The frequencies of isolates resistant to ampicillin, amoxicillin-clavulanate, ciprofloxacin, and gentamicin and also the prevalence of multiresistant strains were greater in surface waters, compared to wastewaters. The Int1I gene was detected in 21.7 % waterborne Enterobacteriaceae. A decrease in coliform counts and intI1-bearing cells, but a general increase in AMR and multiresistant bacteria, occurred during the wastewater treatment. A weak positive correlation was found between multidrug resistance int1I carriage in wastewater effluent but no sufficient evidence of a linkage between phenotypic AMR and int1I, overall. The presence of class 1 integron can be associated with anthropogenic influence, but the simple detection of intI1 gene cannot explain the complex antibiotic resistance phenotype.

Prevalence of class 1 and 2 integrons in multi-drug resistant Escherichia coli isolated from aquaculture water in Chaharmahal Va Bakhtiari province, Iran

Background: Integrons play important role in the spread and maintenance of antimicrobial resistance among strains of Escherichia coli (E. coli) and other species of Enterobacteriaceae. This study investigated the prevalence of class 1 and 2 integrons among E. coli strains isolated from aquaculture water of fish fields in Iran. Methods: One hundred and fifty water samples from different geographical regions in Chaharmahal Va Bakhtiari province were examined over a 2 months period. Isolation was through culture and biochemical tests. Integrons were identified through polymerase chain reaction (PCR) using oligonucleotide primers specific for class 1 and 2 integrons. Antimicrobial susceptibility testing was carried out using disc diffusion methods. Results: Eighteen percent of the water samples were positive for E. coli. All the strains were multi-drug resistant; 100% to ciprofloxacin, chloramphenicol, gentamycin, ampicillin and tetracycline and least resistant to imipenem (7.2%). Ten (50%) of the most resistant strains were positive for class 1 (40%) and class 2 (10%). Conclusions: Escherichia coli in aquaculture in Iran carried integrons class 1 and 2 which could be of public health concern since they could play a role in the spread and maintenance of antimicrobial resistance among bacterial population in the region and should be constantly monitored.

Occurrence of Classes I and II Integrons in Enterobacteriaceae Collected from Zagazig University Hospitals, Egypt

Integrons are genetic units characterized by the ability to capture and incorporate gene cassettes, thus can contribute to the emergence and transfer of antibiotic resistance. The objectives of this study were: 1) to investigate the presence and distribution of class I and class II integrons and the characteristics of the gene cassettes they carry in Enterobacteriaceae isolated from nosocomial infections at Zagzig University Hospital in Egypt, 2) to determine their impact on resistance, and 3) to identify risk factors for the existence of integrons. Relevant samples and full clinical history were collected from 118 inpatients. Samples were processed; isolated microbes were identified and tested for antibiotic susceptibilities. Integrons were detected by polymerase chain reaction (PCR) and were characterized into class I or II by restriction fragment length polymorphism (RFLP). Integron- positive isolates were subjected to another PCR to detect gene cassette, followed by gene cassette sequencing. Risk factors were analyzed by logistic regression analysis. Seventy-six Enterobacteriaceae isolates were recognized, 41 of them (53.9 %) were integron-positive, and 39 strains carried class 1 and 2 strains carried class II integrons. Integrons had gene cassettes encoding different combinations and types of resistance determinants. Interestingly, blaOXA129 gene was found and ereA gene was carried on class I integrons. The same determinants were carried within isolates of the same species as well as isolates of different species. The presence of integrons was significantly associated with multidrug resistance (MDR). No risk factors were associated for integron carriage. We conclude that integrons carrying gene cassettes encoding antibiotic resistance are significantly present among Enterobacteriaceae causing nosocomial infection in our hospital. Risk factors for acquisition remain to be identified.

Occurrence of integrons and antimicrobial resistance genes among clinical isolates of Enterobacter spp. from hospitals of Tehran

Iranian Journal of Public Health, 2014

To determine the occurrence of antimicrobial resistance genes and role of integrons among 135 antimicrobial-resistant Salmonella enterica from Brazil. Methods: The presence of antimicrobial resistance genes, class 1 and 2 integrons and gene cassettes was analysed by PCR and sequencing. The genetic location of class 1 integrons was determined in 25 isolates by hybridization and plasmid transfer experiments. Results: Fifty-five of the isolates were positive for class 1 integrons. Integron-positive isolates represented 17 different serovars and were mainly from human (n = 28) and animal (n = 13) sources. The gene cassette arrangements could be determined in 51 of the positive isolates, which harboured one [dfrA22, aadA1 or orf3 (putative trimethoprim resistance)], two [aadA1-dfrA1, aac(6 0)-Ib-orf1 (unknown function) or aacA4-aadA1], three [dfrA15b-cmlA4-aadA2, orf2 (unknown function)-dfrA5-orfD] or four [orf4-aacA4bla OXA-30 (interrupted by an IS1 element)-aadA1] cassettes in their variable region. Only one isolate harboured a class 2 integron with the gene cassette array dfrA1-sat-aadA1. Several integron unrelated resistance genes were also detected in the isolates. Sulphonamide resistance was primarily mediated by sul2 and sul3, tetracycline resistance by tet(B) and tet(A), chloramphenicol resistance by catA1, streptomycin resistance by strA and ampicillin resistance by bla TEM. bla CTX and bla CMY-2 were found in cephalosporin-resistant isolates. Mating and hybridization experiments demonstrated that a high-molecular-weight plasmid mediated the gene transfer of integrons and additional resistance determinants. Conclusions: The present study revealed that integron-mediated resistance genes contributed to the multiresistance phenotype observed in the isolates, but most resistance genes were located outside the integron structure, as independent genes. However, they might be located on the same conjugative plasmid.

Antimicrobial resistance of enteric bacteria isolated from a sewage treatment plant

O presente trabalho analisou a freqüência de bactérias em uma estação de tratamento de esgoto em Porto Alegre (RS, sul do Brasil). Amostras de água foram coletadas cada três meses, no período de julho/1997 a junho/1998, do afluente, na lagoa de estabilização, na lagoa de maturação e no efluente da estação. Também foram coletadas amostras de dois locais fora da estação (71a e 71b). A resistência a antimicrobianos foi determinada pelo teste de difusão em ágar Muller-Hinton e 18 antimicrobianos diferentes foram testados. Entre as 839 cepas isoladas, Enterobacter sp (n=348) e Escherichia coli (n=210) foram as mais observadas, seguido por Proteus sp (n=66), Serratia (n=53), Citrobacter (n=40), Klebsiella (n=37), Yersinia sp (n=32), Shigella (n=28) e Salmonella (n=25). Uma redução no número de bactérias totais e cepas resistentes a antimicrobianos (cr), pode ser observada do afluente (4,69x10 4 ufc/mL; 154 cr) ao efluente (7,94x10 3 ufc/mL; 49 cr) da estação de tratamento. O número de bactérias resistentes nos pontos 71a e 71b foram tão altos quanto no afluente. O número de cepas multiresistentes também reduziu significativamente do afluente para o efluente. O presente estudo demonstrou que o sistema de tratamento de esgoto estudado é eficiente na redução de enterobactérias e da quantidade de bactérias resistentes a antimicrobianos, contribuindo para um menor lançamento de bactérias multiresistentes no ambiente. Palavras chave: enterobactérias, esgoto, resistência antimicrobiana, ambiente aquático