OJMM  Vol.3 No.3 , September 2013
Plasmid Profiling and Curing Analysis of Fluoroquinolone Multidrug Resistant Pseudomonas aeruginosa in Benin City, Nigeria
ABSTRACT
Aim: To determine the Plasmid DNA profile of the multidrug resistant strains of Pseudomonas aeruginosa in the clinical isolates. Materials and Methods: Of the 150 clinical samples (Ear swab, Urine, Wound swab, Sputa and Semen) received at Lahor Research Laboratory and Medical center in Benin City, between January 2010 and December 2012, 36 (24%) yielded significant growth of P. aeruginosa. Samples were cultured on MacConkey and Blood agar. Clinical isolates were identified using standard method. Antibiotics susceptibility test employing agar disc diffusion method was used. Clinical isolates were subjected to Plasmid DNA profiling and curing test was carried out at Lahor Molecular Laboratory. This was followed by a post plasmid curing susceptibility test. Agarose gel electrophoresis was carried out to separate the Plasmid DNA using standard method. Bands were visualized using UV illuminator. Results: Wound swabs had the highest numbers of clinical isolates of P. aeruginosa (55.6%) followed by Urine, Semen, Sputa and Ear swab (19.4%, 11.0%, 8.3%, and 5.6%) respectively. Before the isolates were cured of their plasmid, 39% of the P. aeruginosa strains were found to be resistant to Ciprofloxacin (CPX), 47%, Ofloxacin (OFX), 44% Pefloxacin (PEF) and 56% Sparfloxacin (SPX). After plasmid curing, the new antibiogram of the isolates showed that some clinical isolates that hitherto were resistant to a given Fluoroquinolone became susceptible, 36% to CPX, 12% to OFX, 12.5% to PEF and 15% to SPX. Agarose gel electrophoresis carried out on the Plasmid DNA revealed that there was detectable Plasmid DNA in 13.9% of the clinical isolates analyzed. Conclusion: There is an alarming increase of clinical infections caused by multidrug resistant strains of P. aeruginosa.13.9% of the multidrug resistance strains of P. aeruginosa in Benin City were Plasmid mediated. Treatment should be based on current Laboratory Susceptibility Test results of the isolates.

Cite this paper
F. Ehiaghe, I. Ehiaghe, D. Agbonlahor, F. Oviasogie, S. Etikerentse, R. Nwobu, B. Akinshipe, A. Ikusemore, S. Aladenika and F. Enwa, "Plasmid Profiling and Curing Analysis of Fluoroquinolone Multidrug Resistant Pseudomonas aeruginosa in Benin City, Nigeria," Open Journal of Medical Microbiology, Vol. 3 No. 3, 2013, pp. 201-205. doi: 10.4236/ojmm.2013.33030.
References
[1]   K. J. Ryan and C. G. Ray, “Sheris Medical Microbiology,” 4th Edition, McGraw Hill, New York, 2004.

[2]   A. Balcht and R. Smith, “Pseudomonas aeruginosa: Infection and Treatment,” Informa Healthcare, 1994, pp. 83-84.

[3]   K. Mathee, G. Narasimhan, C. Valdes, X. Qiu, J. M. Metewish, M. Koehrsen, A. Rokas and C. N. Yandava, “Dynamics of Pseudomonas aeruginosa Genome Evolution,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, No. 8, 2008, pp. 3100-3105. doi:10.1073/pnas.0711982105

[4]   K. Poole, “Efflux Mediated Multiresistance in Gram-Negative Bacteria,” Clinical Microbiology and Infection, Vol. 10, No. 1, 2004, pp. 12-26. doi:10.1111/j.1469-0691.2004.00763.x

[5]   V. Alan, M. Jesus, and W. J. Rober, “The Role of Plasmid in Phytopathogenic Bacteria: Mobile Arsenals,” Journal of General Virology, Vol. 147, No. 4, 2001, pp. 4763-4780.

[6]   N. Datta and V. M. Hughes, “Plasmid of the Same Inc. Group in Enterobacteria before and after the Medical Use of Antibiotics,” Nature, Vol. 306, No. 5943, 1983, pp. 616-613. doi:10.1038/306616a0

[7]   S. T. Cowan and S. Steel, “Manual for the Identification of Medical Bacteria,” 3rd Edition, Cambridge University Press, Cambridge, 1974.

[8]   Clinical Laboratory Standard Institute, “Performance Standard for Antimicrobial Disk Susceptibility Tested: Approved Standard,” 2006.

[9]   T. J. Sijhary, M. L. Bermann and L. W. Enquist, “Experiments with Gene Fusions,” Cold Spring Habor Laboratory Press, New York, 1984.

[10]   H. Bimboim and J. Doly, “A Rapid Alkaline Extraction Procedure for Screening Recombinant Plasmid DNA,” Nucleic Acids Research, Vol. 7, No. 6, 1979, pp. 1513-1523. doi:10.1093/nar/7.6.1513

[11]   J. A. Meyers, D. Sanchez, L. P. Elwell and S. Falkow, “Simple Agarose Gel Electrophoresis Method for the Identification and Characterization of Plasmid Deoxyribonucleic Acid,” Journal of Bacteriology, Vol. 127, No. 3, 1976, pp. 1529-1537.

[12]   K. P. Ranjan, N. Ranjan, K. S. Bansal and P. R. Arora, “Prevalence of P. aeruginosa in Most Operative Wound Infection in a Referral Hospital in Haryana,” Indian Journal of Laboratory Physicians, Vol. 2, No. 2, 2010, pp. 74-77. doi:10.4103/0974-2727.72153

[13]   N. M. R. Bojary and M. Hajia, “Multidrug Resistant P. aeruginosa Strains in Tehran Reference Burn Hospital, Tehran, Iran,” African Journal of Microbiology Research, Vol. 6, No. 7, 2012, pp. 1393-1396.

[14]   M. Hajia, M. Qorbanalizadehgan, M. Rahbar and M. Izadi, “Laboratory Evaluations of Iranian Commercially Provided Antibiotic Disks with Conventional E-Test for Susceptibility Testing in Three Most Isolated Multi-Drug Resistant Organisms,” Internet Journal of Microbiology, Vol. 5, 2008, pp. 23-28.

[15]   M. Rahbar, H. Mehragan and N. Hajiali-Akbori, “Prevalence of Drug Resistance in Non fermenter Gram Negative Bacilli,” Iranian Journal of Pathology, Vol. 5, 2010, pp. 90-96.

[16]   R. A. Bonomo and D. Szabo, “Mechanisms of Multidrug Resistance in Acinetobacter Species and Pseudomonas aeruginosa,” Clinical Infectious Diseases, Vol. 43, No. 2, 2006, pp. 49-56. doi:10.1086/504477

[17]   G. F. Weldhagen, L. Poirel and P. Nordmann, “Ambler Class A Extended Spectrum β-Lactamases in Pseudomonas aeruginosa:novel developments and Clinical Impact,” Antimicrobial Agents and Chemotherapy, Vol. 47, No. 8, 2003, pp. 2385-2392. doi:10.1128/AAC.47.8.2385-2392.2003

[18]   T. R. Walsh, M. A. Toleman, L. Poirel and P. Nordmann, “Metallo—b-Lactamases: The Quiet before Storm? Clinical Microbiology Reviews, Vol. 18, No. 2, 2005, pp. 306-325. doi:10.1128/CMR.18.2.306-325.2005

[19]   G. Agarwal, A. Kapil, S. K. Kabra, B. K. Das and N. Dwivedi, “Characterization of Pseudomonas Aeruginosa Isolated from Chronically Infected Children with Cystic Fibrosis in Indian,” BMC Microbiology, Vol. 5, No. 1, 2005, pp. 43-47. doi:10.1186/1471-2180-5-43

[20]   H. Y. Chen, K. Yunn and D. M. LiverMore, “Mechanism of Resistance to Beta-Lactam Antibiotics among Pseudomonas Aeruginosa Isolates Collected,” Medical Microbiology, Vol. 43, No. 4, 1995, pp. 300-309.

[21]   S. Rahmatis, S. Yang, A. L. Davidson and E. L. Zechiedrich, “Control of the Acr AB Multi-Drug Efflux Pump by Quorum Sensing Regulator,” Molecular Microbiology, Vol. 43, No. 3, 2002, pp. 677-850. doi:10.1046/j.1365-2958.2002.02773.x

[22]   M. Shadid and N. Malik, “Plasmid Mediated Amikacin Resistance in Clinical Isolate of Pseudomonas aeruginosa,” Indian Journal of Medical Microbiology, Vol. 22, No. 3, 2004, pp. 182-184.

[23]   R. C. Clowers, “Molecular Structure of Bacterial Plasmids,” Bacteriological Reviews, Vol. 36, No. 3, 1972, pp. 361- 405.

 
 
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