Departamento de Microbiologia, Imunologia e Parasitologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500. Cidade Baixa, Porto Alegre, RS, 90050-170, Brazil.
Antonie Van Leeuwenhoek. 2012 Jun;102(1):73-81. doi: 10.1007/s10482-012-9714-2. Epub 2012 Mar 1.
The aim of the present study was to investigate the resistance profile, to detect the presence of beta-lactam resistance genes, phenotypic expression of efflux pump systems and class 1 integrons in Pseudomonas spp. strains obtained from untreated hospital effluents. Effluent samples were collected from four hospitals in Porto Alegre, RS, Brazil. Pseudomonas were isolated on MacConkey agar plates and the identification was confirmed by 16S rRNA PCR and biochemical tests. Susceptibility testing was determined by disk-diffusion method using 11 different beta-lactams and MIC assays were performed on isolates resistant to imipenem and ceftazidime. The beta-lactamase genes bla (IMP), bla (VIM), bla (SPM-1), bla (OXA-23-like), bla (OXA-24-like), bla (OXA-51-like) and the intl1 gene from class 1 integron were analysed by PCR. One hundred and twenty-four isolates were recovered and the most common species was Pseudomonas pseudoalcaligenes. The resistance found among the isolates was considered high, 62 (50%) isolates were multiresistant. No isolate carrying the beta-lactamase genes tested was found among the strains. Seven isolates showed reduction of MIC for imipenem and ceftazidime in the presence of cyanide m-chlorophenylhydrazone, indicating the hyper expression of efflux pumps. From the 124 isolates, 52 (41.9%) were identified as carrying the class 1 integron gene, intI1. Untreated hospital effluents could be a source of environmental contamination due to discharge of antimicrobial resistant bacteria which can carry integron class 1 and act as a reservoir of resistance genes and have efflux pump systems.
本研究旨在调查未处理医院污水中分离的铜绿假单胞菌的耐药谱、β-内酰胺耐药基因、外排泵系统表型表达和 1 类整合子的存在情况。污水样本取自巴西阿雷格里港的四家医院。在麦康凯琼脂平板上分离铜绿假单胞菌,并通过 16S rRNA PCR 和生化试验确认鉴定。采用 11 种不同的β-内酰胺类药物进行纸片扩散法药敏试验,对耐亚胺培南和头孢他啶的分离株进行 MIC 测定。采用 PCR 方法检测β-内酰胺酶基因 bla(IMP)、bla(VIM)、bla(SPM-1)、bla(OXA-23 样)、bla(OXA-24 样)、bla(OXA-51 样)和 1 类整合子的 intl1 基因。共回收 124 株分离株,最常见的菌种是 Pseudomonas pseudoalcaligenes。分离株的耐药性较高,62 株(50%)为多药耐药。在所检测的菌株中未发现携带β-内酰胺酶基因的菌株。7 株分离株在氰化 m-氯苯腙存在下,对亚胺培南和头孢他啶的 MIC 降低,表明外排泵的过度表达。在 124 株分离株中,有 52 株(41.9%)携带 1 类整合子基因 intI1。未处理的医院污水可能是由于排放具有整合子 1 类的耐药菌而导致环境污染的来源,这些耐药菌可作为耐药基因的储存库,并具有外排泵系统。