[Correlation between multiple antibiotic resistance and heavy-metal tolerance among some E.coli strains isolated from...

Self-transmissible plasmids conferring multiple antibiotic resistance are wide-spread in coliforms populations. In soil and water, multiple antibiotic resistance is clearly associated with resistance/tolerance to heavy-metals (Hg2+, Cu2+, Pb2+, Zn2+, Ca2+). For different genera the genes for heavy-metals resistance are often plasmid encoded. Since these genes are clustered on the same plasmids, heavy-metals and drugs are environmental factors which exert a selective pressure for the populations of these plasmid-harboring bacteria. The aim of this preliminary study was to find possible correlation between resistance genotype determined by genetic analysis and antibiotic and heavy-metal resistance patterns of 12 E. coli strains isolated from chronically polluted waters. Antimicrobial susceptibility testing was performed for ampicillin, tetracycline, gentamycin, kanamycin, chloramphenicol, ceftazidime and cefotaxime by standard disk diffusion Kirby-Bauer method following NCCLS recommendations. These antibiotics were chosen because of their wide-spread use and importance in the treatment of Gram-negative bacterial infections. MICs values of antibiotics and heavy-metals were determined by dilution method in Mueller-Hinton broth using an inoculum of about 1-2 x 10(8) CFU/ml. The concentration range for antimicrobials and heavy-metals salts (CuSO4, CdCl2, Co(NO3)2, Cr(NO3)3, HgCl2, NiCl2 and ZnSO4) was 0.06-64 [symbol: see text] g/ml, 0.5-256 [symbol: see text] g/ml respectively. Plasmid DNA was isolated from E. coli strains by an alkaline lysis. Genetic characterization was performed by agarose gel electrophoresis and spectrophotometric analysis. All strains are multiple antibiotic resistant, 16% of them being resistant to 3, 4 and 6 antibiotics, 32% to 5 and 8% to all 7 antibiotics, respectively. Multiple tolerance to high levels of Cd2+, Cu2+, Cr3+ and Ni2+ was common among multiple antibioresistant strains. Screening for plasmids relieved the presence of several plasmids ranging from 3.8 kpb to more than 50 kpb. The phenotypic data shows the direct association between multiple antibiotic and heavy-metal resistance for E. coli strains in polluted water. Electrophoretic patterns analysis reflects the high incidence and diversity of analyzed plasmids.