Determination of Plasmid Profile Multiple Antibiotics Resistant Pseudomonas aeruginosa Isolated from Clinical Specimens in Healthcare Facilities across Anambra and Imo States Nigeria

Seraphim Chinyere Ifegbuike

Department of Medical Laboratory Science, Faculty of Health Sciences, Imo State University, Owerri, Nigeria.

Godwin Chukwuemeka Ojiegbe

Department of Medical Laboratory Science, Faculty of Health Sciences, Imo State University, Owerri, Nigeria.

Henry Chidozie Amah

Department of Medical Laboratory Science, Faculty of Health Sciences, Imo State University, Owerri, Nigeria.

Chizaram Winners Ndubueze *

Department of Medical Laboratory Science, Faculty of Health Sciences, Imo State University, Owerri, Nigeria.

Joy Nkeiruka Dike-Ndudim

Department of Medical Laboratory Science, Faculty of Health Sciences, Imo State University, Owerri, Nigeria.

Emeka Simon Anikwo

Department of Human Physiology, Faculty of Basic Medical Sciences, Imo State University, Owerri, Nigeria.

*Author to whom correspondence should be addressed.


Abstract

The emergence and dissemination of antibiotic resistant plasmids of P. aeruginosa is posing a major public threat and huge concern in hospital facilities. This study was done across Anambra and Imo States with a total number of 100 P. aeruginosa isolates, 50 from each State to determine the plasmid profile of multidrug resistant isolates. Methods of re-identifying P. aeruginosa were based upon cultural methods coupled with biochemical tests. To study the susceptibility of these isolates using disk diffusion method, seven (7) antibiotics were used namely: Piperacillin–tazobactam (100/10 µg), Ceftazidime (30 µg), Amikacin (30 µg), Ciprofloxacin (5 µg), Cefuroxime (30 µg), Ceftriazone (30 µg) and Gentamicin (10 µg). Plasmid extraction was done using alkaline lysis method after growing isolates resistant to more than four antibiotics on a nutrient broth. Agarose gel electrophoresis of plasmid DNA was carried out on 2% agarose gel slab in 1X TAE buffer. The results show that, the nutrient agar at 37˚C aerobically, P. aeruginosa isolates were recovered, which produced greenish-yellow pigment colonies, oxidase was positive and negative for gram stain. In Anambra, the result showed 100% resistance to Cefuroxime, Ceftazidime and Ciprofloxacin, 90% and 86% for Ceftriaxone and Piperacillin-tazobactem, 48% and 40% to Gentamicin and Amikacin respectively. Whereas in Imo state, the result showed 100% resistance to Ceftazidime and Ciprofloxacin, 80% to Ceftriaxone, Cefuroxime and Piperacillin-tazobactem while the least resistance was seen in Amikacin, and Gentamicin. Plasmid size ranging from 100bp to >1000bp was detected from most of the multidrug resistant isolates. Not all the isolates with multidrug resistance were found to possess plasmids. It can be seen from this study that multidrug resistance in P. aeruginosa is not strictly plasmid-dependent (mediated).

Keywords: Plasmid, antibiotics resistance, Pseudomonas aeruginosa, clinical isolates, Imo, Anambra, Nigeria


How to Cite

Chinyere Ifegbuike, Seraphim, Godwin Chukwuemeka Ojiegbe, Henry Chidozie Amah, Chizaram Winners Ndubueze, Joy Nkeiruka Dike-Ndudim, and Emeka Simon Anikwo. 2021. “Determination of Plasmid Profile Multiple Antibiotics Resistant Pseudomonas Aeruginosa Isolated from Clinical Specimens in Healthcare Facilities across Anambra and Imo States Nigeria”. Journal of Advances in Medical and Pharmaceutical Sciences 23 (11):17-28. https://doi.org/10.9734/jamps/2021/v23i1130268.

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