Evaluation of antimicrobial combination therapy options for the management of integron-mediated multidrug resistance in enterococcus species and acinetobacter baumannii from aquatic environment in the Eastern Cape Province, South Africa
- Authors: Ola, Adeniji Oluwaseun
- Date: 2022-04
- Subjects: Enterococcus , Aquatic biodiversity , Acinetobacter
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/27776 , vital:69467
- Description: Infectious diseases caused by multidrug-resistant MDR pathogens, such as Acinetobacter baumannii and Enterococcus spp., is an increasing worldwide problem. For treating these diseases, antibiotics are usually the first choice. But organisms develop resistance as a result of drug abuse, continuous use of antibiotics and release of antibiotics into the environment. These have prompted MDR's development, making even the most active drugs ineffective. Transposons, plasmids and integrons are the most effective mobile genetic elements that promote acquisition and spread of resistance determinants. Integrons carrying various arrays of resistance gene cassettes are principally helpful for epidemiological studies of these disease-causing organisms. Alternative treatments, such as using drugs in combination or with adjuvants and nanoparticles therapy, have been documented. Nanoparticles have the potential requirements for qualifying as antibacterial agents. In addition to their antimicrobial activities, nanoparticles can be used together with antibiotics for more enhanced antimicrobial activity In this study, Acinetobacter baumanni, Enterococcus faecium and Enterococcus faecalis were recovered from the aquatic environment in the Eastern Cape Province, South Africa with a standard microbiological method. Their antibiotic sensitivity testing was carried out using Kirby Bauer disk diffusion and microdilution methods. A high occurrence of class-1 integrons was discovered in MDR A. baumanni, with the internal variable containing aadA1, aadA5 and aadA2 genes, which confer resistance for streptomycin and spectinomycin, aac 6Ib for amikacin/ tobramycin and dfrA17 genes for trimethoprim. Similarly, class1 integron was detected in Enterococcus, without the presence of gene cassette. The checkerboard assay and time-kill assay were used to test for the effect of the combination of the antibiotic. The impact of colistin combined with quinolones (ciprofloxacin) with the Fractional inhibitory concentration index FICs 0.31 indicated synergistic effects against MDR A baumanni. However, when colistin was combined with meropenem and ceftazidime, additive effects with FIC, ranging from 0.52 to 1 were observed. In addition, a combination of gentamicin MIC 4 μgml with vancomycin MIC 256 μgml antibiotics against vancomycin-resistant Enterococcus faecalis showed antibacterial activity. In contrast, the combination of ciprofloxacin 1 μgml with Ampicillin 16 μgml antibiotics against Enterococcus faecalis showed a bacteriostatic effect. The initial inoculum declined by 100 percentage when gentamicin was combined with vancomycin at a concentration of 4 and 128 μgml MIC respectively, for about 2 h following the treatment for MDR E. faecium. Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction ERIC PCR analyses of the studied pathogens revealed great genetic diversity, suggesting the various sources of environmental contamination. Silver nanoparticles AgNPs and zinc oxide nanoparticles ZnO NPs were chemically synthesized using a precipitation method and characterized using energy dispersive Xray analysis EDX, scanning electron microscopy SEM, Fourier transforms infrared spectroscopic analysis FTIR and transmission electron microscopy TEM. The characterization results showed the synthesis of 43.37 nm and 21.03 nm nanoparticles of zinc oxide and silver origins, correspondingly, with distinct morphology, as revealed in TEM. The size, stability and functional groups of the nanoparticles produced were revealed using EDX and FTIR. Zinc oxide nanoparticles were cytotoxic against Vero cell lines at the tested concentrations, whereas AgNPs had no cytotoxic effect at lower concentrations. Acinetobacter baumanni, Enterococcus faecium and Enterococcus faecalis were with MIC in the range of 0.0390.078mgml for silver nanoparticles. Zinc oxide nanoparticles were explicitly active against Enterococcus species gram-positive with MIC of 1.25 5 mgml. Both studied nanoparticles exhibited profound synergistic and additive activities against all the investigated MDR pathogens. These findings demonstrate good antibacterial potential of the nanoparticles against drug-resistant strains and open a new arena of antimicrobials for medical treatment. , Thesis (MSc) -- Faculty of Science and Agriculture, 2022
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- Date Issued: 2022-04
Enterococcus pathotypes as reservoirs of antibiotic resistance determinants in the Kat River and Fort Beaufort abstraction waters
- Authors: Ntloko, Phindiwe
- Date: 2014
- Subjects: Enterococcus , Drug resistance
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11290 , http://hdl.handle.net/10353/d1019821 , Enterococcus , Drug resistance
- Description: In this study, 400 presumptive Enterococcus isolates previously recovered from Kat River and Fort Beaufort Abstraction water dam were subjected to molecular confirmation and pathotyping. Two hundred and seventy-four (68%) of these isolates were confirmed to be enterococci species. Confirmations studies were polymerase chain reaction (PCR) based, using enterococci specific primers targeting the tuf gene. The confirmed enterococci isolates were further differentiated into their pathotypes, the targets of which were: E. faecalis, E. avium, E. hirae, E. casseliflavarus and E. gallinarum using well documented species specific primer sequences. E. faecalis accounted for 20% of the isolates, followed by E. avium (16%), E. hirae (13%), E. casseliflavarus (5%) and E. gallinarum (3%). Furthermore, all the confirmed isolates were analysed for antibiotic susceptibilities using a panel of nine different antibiotics, namely vancomycin, linezolid, ciprofloxacin, ampicillin, gentamicin, chloramphenicol, tetracycline, erythromycin, penicillin, and those that were resistant were assayed for the presence of relevant antibiotic resistance genes. All the 274 isolates were found to harbour vanA resistance gene confirming their phenotypic resistance to the vancomycin. Similarly, 60% (109/180) of the isolates showed phenotypic resistance to erythromycin which was further confirmed by the presence of ermA genes in these isolates. The presence of antibiotic resistant bacteria in surface waters poses a risk to public health.
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- Date Issued: 2014