Colistin Resistance Profiles, Molecular Investigation of mcr-1 and mcr-2 Plasmid Genes and Investigation of Carbapenemase Production in Pseudomonas and Acinetobacter Strains

Background and Purpose: The reintroduction of colistin as a last resort treatment against multi-resistant Gram-negative bacilli, is currently challenged by the emergence of colistin-resistant bacteria. The aim of this study was to assess the susceptibility of Pseudomonas and Acinetobacter strains to colistin, to identify carbapenemase production, and to investigate the plasmid genes involved in colistin resistance and carbapenemase production. Methodology: In order to establish the susceptibility profiles of 17 strains of Pseudomonas and Acinetobacter to colistin, their Minimum Inhibitory Concentrations (MICs) were determined using the liquid microdilution method. The possible production of carbapenemases was investigated with the modified Carbapenem Inactivation Method (mCIM). The search for genes encoding carbapenemases (blaOXA, blaIMP, blaCarba) and those responsible for plasmid resistance to colistin (mcr-1 and mcr-2) was performed by conventional PCR. Results and Conclusion: Ninety-four percent (94%) (16/17) of the strains were resistant to colistin. Intraspecies distribution was 50% (8/16), 31% (5/16), 13% (2/16) and 6% (1/16) for Acinetobacter baumannii, Pseudomonas aeruginosa, Pseudomonas luteola, and Pseudomonas fluorescens, respectively. Twenty-nine percent (29%) (6/17) of the strains produced carbapenemases. No mcr-1 and mcr-2 plasmid genes were detected. On the other hand, 17.6% (3/17) of the strains possessed the carbapenemase genes distributed as follows: Carba type (60%), OXA type (40%) and IMP type (0%). The results of this study highlight a high resistance to colistin in strains belonging to the genera Acinetobacter and Pseudomonas, and some of these strains produce carbapenemases.

Molecular Prevalence and Epidemiological Characteristics of Diarrheagenic <i>E. coli</i>in Children under 5 Years Old in the City of Koula-Moutou, East-Central Gabon

Background and Purpose: Diarrhoeagenic E. coli (DEC) is one of the germs responsible for childhood diarrhea in developing countries. This study aims at determining the prevalence of the five main pathotypes of DEC isolated from faeces of children under five years old with diarrhea or not, living in the city of Koula-Moutou. Methodology: Isolates of E. coli were phenotypically screened on chromIDTM agar and molecularly by multiplex PCR to detect the presence of enteroaggregative E. coli (EAEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enterohemorragic E. coli (EHEC) and enteroinvasive E. coli (EIEC). The evaluation of their sensitivity to 12 β-lactam antibiotic molecules was carried out by Kirby Bauer method. This method has also made it possible to characterize phenotypically the different β-lactamases produced. Results and Conclusion: Overall, at least one DEC pathovar was detected in the 63 E. coli strains with phenotypic and molecular frequencies of 63.5% and 68.5% respectively. Thus, ETEC (28.3%) and EHEC (28.3%) were the most frequent DEC in diarrheal isolates. ETEC/EHEC hybrid was recorded in both groups with rates of 7.5% in diarrheal cases and 10.0% for controls. The results showed produced carbapenemase type β-lactamases (31.7%), followed by ESBL (24.4%) and few produced high level penicillinases (4.9%). The DEC, in particular ETEC and EHEC are most likely the epidemiological agents responsible for childhood diarrhea in this study.