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artigo originaL / research report / artícuLo
Detection of antimicrobial-resistant gram-negative bacteria
in hospital effluents and in the sewage treatment station
of Goiânia, Brazil
Detección de bacterias gram negativas resistentes a antimicrobianos en efluentes hospitalarios y en la
estación de tratamiento de aguas residuales de Goiânia, Brasil
Detecção de bactérias gram-negativas antimicrobiano-resistentes em efluentes de hospitais e na estação
tratamento de esgoto de Goiânia, Brasil
Aline Cristina Batista Resende*
Renata de Bastos Ascenso Soares**
Daniela Braz dos Santos***
Edlaine Rodrigues Montalvão****
José Rodrigues do Carmo Filho*****
ABStrAct: The emergence of antimicrobial-resistant genes and the indiscriminate use of antibiotics contribute to the dissemination of resis-
tant pathogens in the environment. The objective of the present study was to isolate Pseudomonas aeruginosa, Acinetobacter spp., Klebsiella pneumoniae and Escherichia coli from the sewage effluents of 10 hospitals located in Goiânia, Brazil, and from the sewage treatment station of the city, to determine their susceptibility profile and investigate their resistance mechanisms. The isolates from water samples were identified by biochemical tests and confirmed using API 20E (BioMerieux). Susceptibility profiling was performed by disc diffusion in accordance with the methodology established by the National Committee for Clinical Laboratory Standards. Extended-spectrum β-lactamase (ESBL) detection was carried out by the disk approximation method using phenotypic tests. Sixty-seven microorganisms were isolated and identified, including E. coli 10 (14,92%), K. pneumoniae 10 (14,92%), P. aeruginosa 3 (4,47%) and A. baumannii 1 (1,49%). Of the E. coli strains, 100% were resistant to aztreonam, 40% to ampicillin, 30% to piperacillin, 20% to ciprofloxacin and 10% to gentamicin. None of the bacterial strains produced ESBL or carbapenems. Of the P. aeruginosa strains, 100% were resistant to ampicillin-sulbactam, while 100% had intermediate resistance to gentami- cin. Strains of K. pneumoniae were resistant to ampicillin (70%) and to piperacillin (20%); additionally, 50% showed intermediate resistance to piperacillin. Total resistance was not found in any of the isolates of A. baumannii, which showed intermediate resistance to aztreonam and ceftriaxone. Overall, resistance rates were low in the isolates of E. coli, P. aeruginosa, K. pneumoniae and A. baumannii.
KeyworDS: Antimicrobial-resistant gram-negative bacteria. Dissemination of resistant pathogens – environment. Hospitals.
reSumen: La aparición de genes resistentes a antimicrobianos y la utilización indiscriminada de antibióticos contribuyen a la difusión de pa-
tógenos resistentes en el ambiente. El objetivo de este estudio fue aislar Pseudomonas aeruginosa, Acinetobacter spp., Klebsiella pneumoniae y Escherichia coli en efluentes de aguas residuales de 10 hospitales situados en Goiânia, Brasil, y de la estación de tratamiento de aguas residuales de la ciudad intentando determinar su perfil de susceptibilidad e investigar a sus mecanismos de resistencia. Los aislados de muestras de agua fueron identificados de promedio pruebas bioquímicas y confirmados utilizando API 20E (BioMerieux). El perfil de susceptibilidad fue realizado por difusión de disco de acuerdo con la metodología establecida por el National Committee for Clinical Laboratory Standards. La detección de beta-lactamasas de espectro extendido (ESBL) fue realizada de promedio el método de aproximación de discos utilizando pruebas fenotípicas. Sesenta y siete microorganismos fueron aislados e identificados, incluyendo E. coli 10 (14,92%), K. pneumoniae 10 (14,92%), P. aeruginosa 3 (4,47%) y A. baumannii 1 (1,49%). Las cepas de Escherichia Coli fueran 100% resistentes a aztreonam, 40% a ampicilina, 30% a piperacilina, 20% a ciprofloxacino y 10% a gentamicina. Ningunas de las cepas bacterianas produjeron ESBL o carbapenems. Las cepas de P. aeruginosa fueran 100% resistentes a ampicilina-sulbactam, mientras 100% presentaran una resistencia media a gentamicina. Las cepas de K. pneumoniae fueran resistentes a ampicilina (el 70%) y a piperacilina (el 20%); además, el 50% presentaran resistencia media a piperacilina. La resistencia total no fue encontrada en aislados de A. baumannii, que presentaran resistencia media a aztreonam y ceftriaxona. En términos globales, las tajas de resistencia fueran bajas en los aislados de Escherichia Coli, P. aeruginosa, K. pneumoniae y A. baumannii.
PAlABrAS llAve: Bacterias gram negativas resistentes a antimicrobianos. Difusión de patógenos resistentes - ambiente. Hospitales.
reSumo: A emergência de genes antimicrobiano-resistentes e o uso indiscriminado de antibióticos contribuem para a disseminação de patógenos
resistentes no ambiente. O objetivo deste estudo foi isolar Pseudomonas aeruginosa, Acinetobacter spp., Klebsiella pneumoniae e Escherichia coli em efluentes de esgoto de 10 hospitais situados em Goiânia, Brasil, e da estação de tratamento de esgoto da cidade, para determinar seu perfil de susceptibilidade e investigar seus mecanismos de resistência. Os isolados das amostras de água foram identificados usando testes bioquímicos e confirmados com API 20E (BioMerieux). O perfil de susceptibilidade foi estabelecido pela difusão de disco de acordo com a metodologia estabelecida pelo National Committee for Clinical Laboratory Standards. A detecção de Beta-Lactamase de Espectro Estendido (ESBL) foi realizada pelo método de aproximação de discos usando testes fenotípicos. Sessenta e sete microorganismos foram isolados e identificados, incluindo E. coli 10 (14,92%), K. pneumoniae 10 (14,92%), P. aeruginosa 3 (4,47%) e A. baumannii 1 (1,49%). Dentre as cepas de Escherichia Coli, 100% foram resistentes a aztreonam, 40% a ampicilina, 30% a piperacilina, 20% a ciprofloxacina e 10% a gentamicina. Nenhuma das cepas bacterianas produziu ESBL ou carbapenems. Dentre as cepas de P. aeruginosa, 100% foram resistentes a ampicilina-sulbactam, enquanto 100% mostraram resistência média a gentamicina. As cepas de K. pneumoniae foram resistentes a ampicilina (70%) e piperacilina (20%); adicionalmente, 50% mostraram resistência média a piperacilina. Não houve casos de resistência total em alguns dos isolados de A. baumannii, que tiveram resistência média a aztreonam e ceftriaxona. De modo geral, as taxas de resistência foram baixas nos isolados de P. aeruginosa, Escherichia Coli, K. pneumoniae e A. baumannii.
PAlAvrAS-chAve: Bactérias gram-negativas antimicrobiano-resistentes. Disseminação de patógenos resistentes - ambiente. Hospitais.
* Master’s program in Environmental Sciences and Health, Catholic University of Goiás, Goiás, Brazil.
** Professor, Catholic University of Goiás, Goiás, Brazil.
*** Undergraduate, Catholic University of Goiás, Goiás, Brazil.
**** Graduate in Biomedicine, Catholic University of Goiás, Goiás, Brazil.
***** Professor, Catholic University of Goiás, Goiás, Brazil. E-mail: Biomedico47@gmail.com O MundO da Saúde, São Paulo: 2009;33(4):385-391.
Detection of antimicrobiaL-resistant gram-negative bacteria in hospitaL effLuents anD
in the sewage treatment station of goiânia, braziL
Introduction
diffusion test. E. coli ATCC 35218 as Escherichia coli, Klebsiella pneu- moniae, Acinetobacter spp. and E. coli, 8 isolates (36,4%); K. pneumonia, 10 (45,5%); P. aerugi- nosa, 3 (13,6%), one A. bauman- nii, (4,5%) and E. coli, 2 isolates the E. coli isolates to some classes materials and methods
(NCCLS)20. Standard strains Escher- ichia coli ATCC 25922, P. aerugino- (10%) E. coli strains, respectively. sa ATCC 27853, and K. pneumoniae O MundO da Saúde, São Paulo: 2009;33(4):385-391.
Detection of antimicrobiaL-resistant gram-negative bacteria in hospitaL effLuents anD
in the sewage treatment station of goiânia, braziL
and all E. coli isolates were resistant inhibited growth of all the K. pneu- found in the unique strains of A. baumannii isolated in this study. coli isolates that were resistant to The sensitivity profile of the K. pneumoniae isolates showed that sev- Discussion
eracilin (five strains – 50%), and for otics in the environment5,7,17,18,19,28.
table 1. Susceptibility profile of the microorganisms isolated from the sewage systems of 10 hospitals, and the
municipal sewage treatment station of Goiânia city, GO, Brazil p. aeruginosa
K. pneumoniae
a. baumannii
antimicrobials
Abbreviations: AMP - ampicillin, PIP - piperacillin, ATM - aztreonam, PTZ - piperacillin-tazobactam, CAZ - ceftazidime, CTX - cefotaxime, CRO - ceftriaxone, IMP - imipenem, CIP - ciprofloxacin, GEN - gentamicin, MI - amikacin, CPM - cefepime, AST - ampicillin / sulbactam, CPD - cefpodoxime, TIC - ticarcillin. S = sensibility; R = resistance; I = intermediate resistance.
O MundO da Saúde, São Paulo: 2009;33(4):385-391.
Detection of antimicrobiaL-resistant gram-negative bacteria in hospitaL effLuents anD
in the sewage treatment station of goiânia, braziL
It has been shown that E. coli is of E. coli, three of P. aeruginosa, 10 K. pneumoniae and one A. bauman- nii. Resistance of these isolates was selection of resistance of E. coli, and to a greater extent resistance of P. isolated 34 strains of E. coli, 10 of aeruginosa, to ciprofloxacin. Bacteria resistant to tetracycline, including P. aeruginosa, have also been identified Acinetobacter, Alcaligenes, Citrobacter, Enterobacter, Pseudomonas, Serratia, Klebsiella and Proteus, the latter dime consisted of Pseudomonas. As to the others2. In another study, K. pneumoniae, E. coli, E. cloacae, C. fre- undii, Aeromonas spp., S. marcescens, ated with high mortality rates2,38,39.
Citrobacter spp., K. oxytoca and A. cal- coaceticus were detected in samples isms such as Moraxella, Acinetobacter, Flavobacterium, Pseudomonas, Aero- monas, Bacillus, Proteus, Arthrobacter, sistance of P. aeruginosa isolated in Lactobacillus, Klebsiella, Plesiomonas, Serratia, Enterobacter, Staphylococcus and Micrococcus were also found in In the present study, E. coli was aeruginosa. All the isolates had in- resistance to this antimicrobial9,25.
detected the presence of E. coli with them P. aeruginosa, were resistant O MundO da Saúde, São Paulo: 2009;33(4):385-391.
Detection of antimicrobiaL-resistant gram-negative bacteria in hospitaL effLuents anD
in the sewage treatment station of goiânia, braziL
profile of K. pneumoniae, this micro- and identified 30 bacteria (Aeromo- K. pneumoniae infection is less than nas hydrophila¸ Stenotrophomonas maltophilia, Enterobacter spp, E. as- buriae), of which 29 were resistant (15), over 80% of the isolates of E. coli, K. pneumoniae, E. cloacae and P. aeruginosa were susceptible to cip- in which few isolates of E. coli and K. pneumoniae were resistant to this sistant bacteria is a public health issue, our data support the need of against Acinetobacter baumannii15, O MundO da Saúde, São Paulo: 2009;33(4):385-391.
Detection of antimicrobiaL-resistant gram-negative bacteria in hospitaL effLuents anD
in the sewage treatment station of goiânia, braziL
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Recebido em 3 e agosto de 2009
Aprovado em 22 de setembro de 2009
O MundO da Saúde, São Paulo: 2009;33(4):385-391.

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