PCR Analysis of Resistant Bacteria Strains Isolated from River Sokoto, Northwestern Nigeria

PCR Analysis of Resistant Bacteria Strains Isolated from River Sokoto, Northwestern Nigeria

Loading document ...
Page
of
Loading page ...

Author(s)

Author(s): Raji M.I.O., Garba I.

Download Full PDF Read Complete Article

DOI: 10.18483/ijSci.1289 158 480 9-14 Volume 6 - Aug 2017

Abstract

Thirteen resistant bacteria strains from River Sokoto namely Salmonella typhi, Escherichia coli, Shigella flexneri, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus faecalis, Bacillus subtilis, Enterobacter cloacae, Klebsiella oxytoca, Staphylococcus saprophyticus, Providencia rettgeri, Klebsiella pneumonae and Enterobacter aerogenes recovered on Mueller-Hinton agar by disc diffusion method were subjected to PCR (Polymerase chain reaction) analysis to determine their antibiotic resistance genes. Forward and reverse copies of five primers (TEM, spvC, SHV, aacC3 and qnrS) were used in the PCR analysis. Aminoglycoside resistance genes (aacC3) were detected in the majority of the isolates such as E. coli, Shigella flexneri, Enterobacter cloacae, Staphylococcus saprophytica and Enterobacter aerogenes with plasmid numbers ranging from 1 to 4 and molecular weights ranging from 185 bp to >10,200 bp. Virulence resistance genes (spvC) were detected in Salmonella typhi on two plasmids with molecular weights of 571 bp and >10,200 bp while quinolones resistance genes were detected on plasmid numbers ranging from 1 to 3 and molecular weights ranging from 400 bp to 1000 bp in Staphylococcus aureus, Streptococcus faecalis, Bacillus subtilis and Providencia rettgeri. Three different resistance genes namely β-lactam (blaTEM), virulence (spvC) and quinolones (qnrS) on 6 plasmids with molecular weights ranging from 428 bp to 1,200 bp were found in Pseudomonas aeruginosa while two resistance genes (aacC3 and qnrS) on 2 plasmids with molecular weights of 185bp and 428bp were detected in Klebsiella pneumoneae. Non-specific resistance genes alongside with specific resistance genes were however detected in the majority of the isolates. The study revealed that the resistance genes exhibited by resistant bacteria isolates from River Sokoto were mainly virulence, aminoglycoside and quinolones resistant genes. The scientists are therefore challenged on the need for development of new antibiotics to combat the infections caused by these resistant strains.

Keywords

PCR Analysis, Resistant Bacteria, Antibiotic Resistance Genes, Non-Specific Resistance Genes

References

  1. Alzahrani, A.M. and Gherbawy, Y.A. (2011). Antibiotic resistance in Escherichia coli strains isolated from water springs in Al-Ahsa Region. Afr. J. Microbiol. Res., 5(2): 123-130.
  2. Akinyemi, K. O., Oyefolu, A. O. B., Salu, O. B., Adewale, O. A. and Fasure, A. K. (2006). Bacterial pathogens associated with tap and well waters in Lagos, Nigeria. East Cent. Afr. J. Surg. 11(1): 110-117.
  3. Andres, P.; Lucero, C.; Soler-Bistue, A.; Guerriero, L.; Albornoz, E.; Tran, T.; Zorreguieta, A.; Galas, M.; Corso, A. and other authors (2013).Differential distribution of plasmid-mediated quinolone resistancegenes in clinical enterobacteria with unusual phenotypes of quinolonesusceptibility from Argentina.Antimicrob Agents Chemother 57: 2467–2475.
  4. Barza M, Travers K (2002). Excess infections due to antimicrobial resistance: the “Attributable Fraction”, In: The need to Improve Antimicrobial Use in Agriculture: Ecological and Human Health Consequences, M Barza and SL Gorbach (Eds.,), Clin. Infect. Dis., 34: 126-130.
  5. Baquero F, Martinez JL, Canton R (2008). Antibiotics and antibiotic resistance in water environments. Curr. Opin. Biotechnol., 19: 260 - 265.
  6. Belaaouaj, A., Lapoumeroulie, C, Canica, M. M., Vedel, G., Nevot, P., Krishnamoorthy, R. (1994). Nucleotide sequences of the genes coding for the TEM-like /5-lactamases IRT-1 and IRT-2 (formerly called TRI-1 and TRI-2). FEMS Microbiology Letters 120: 75-80.
  7. Carattoli, A. (2003) Plasmid-mediated antimicrobial resistance in Salmonella enteric. Curr.Issues Mol. Biol. 5: 113-122.
  8. Carlson, S.A., Bolton, L.F., Briggs, C.E., Hurd, H.S., Sharma, V.K., Fedorka-Cray, P.J. and Jones, B.D. (1999) Detection of multiresistant Salmonella typhimuriium DT 104 using multiplex and fluorogenic PCR. Molecular and Cellular Probes 13: 213-222.
  9. Cattoir, V., Poirel, L., Rotimi, V., Soussy, C. J. and Nordmann, P. (2007). Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates. J Antimicrob Chemother 60: 394–397.
  10. Deepak, R. N.; Koh, T. H. and Chan, K. S. (2009). Plasmid-mediated quinolone resistance determinants in urinary isolates of Escherichia coli and Klebsiella pneumoniae in a large Singapore hospital. Ann Acad Med Singapore 38: 1070–1073.
  11. Dubey, J.P. (2009) Toxoplasmosis of Animals and Humans, Second ed. CRC Press, Boca Raton, FL.
  12. Hamner, S., Tripathi, A., Mishra, R.K. (2006) The role of water use patterns and sewage pollution in incidence of water-borne/enteric diseases along the Ganges River in Varanasi, India. International Journal of Environmental Health Research, 16 (2):113–132.
  13. Liu, B. T., Liao, X. P., Yue, L., Chen, X. Y., Li, L., Yang, S. S., Sun, J., Zhang, S., Liao, S. D. and Liu, Y. H. (2013). Prevalence of β-lactamase and 16S rRNA methylase genes among clinical Escherichia coli isolates carrying plasmid-mediated quinolone resistance genes from animals. Microb Drug Resist 19: 237–245.
  14. Marti, E., Jofre, J. and Balcazar, J.L. (2013). Prevalence of Antibiotic Resistance Genes and Bacterial Community Composition in a River Influenced by a Wastewater Treatment Plant. PLoS ONE 8(10): http://dx.doi.org/10.1371/journal.pone.0078906
  15. Odeyemi, A.T., Awokunmi, E.E., Adebayo, A.A., Fasoranti, O.F. (2015) Plasmid profile of multi-drug resistance bacteria isolated from available water sources and leachate samples from dumpsite at Ebira communities in Ekiti North Senatorial District, Ekiti State, Nigeria. European Journal of Advanced Research in Biological and Life Sciences. 3(1): 31 – 45.
  16. Olofsson, S. (2006). Relation between dug exposure and selection of antibiotic resistant bacteria. Dissertation presented at the Faculty of Medicine, Upsalla University for the degree of Doctor of Philosophy.
  17. Oyedeji, O., Olutiola, P.O., Owolabi, K.D., Adeojo, K.A. (2011). Multi-resistant faecal indicator bacteria in stream and well waters of Ile-Ife City, Southwestern Nigeria: Public health implications. J. Public Health Epidemiol. 3(8): 371-381.
  18. Park, C. H., Robicsek, A., Jacoby, G. A., Sahm, D. and Hooper, D. C. (2006) Prevalence in the United States of aac(69)-Ib-cr encoding a ciprofloxacin-modifying enzyme. Antimicrob Agents Chemother 50: 3953–3955.
  19. Pitout, J.D.D., Thomson, K.S., Hanson, N.D., Ehrhardt, A.F., Moland, E.S. and Sanders, C.C. (1998) β-lactamases responsible for resistance to expanded-spectrum cephalosporins in Klebsiella pneumoniae, Escherichia coli, and Proteus mirabilis isolates recovered in South Africa. Antimicrobial Agents and Chemotherapy 42: 1350–1354.
  20. Ramirez, M.S.; Traglia, G.M.; Lin, D.L.; Tran, T. and Tolmasky, M.E. (2014) Plasmid-mediated antibiotic resistance and virulence in Gram-negatives: the Klebsiella pneumoniae paradigm. Microbiol Spectr. 2(5): 1–15.
  21. Shaheen, B. W., Nayak, R., Foley, S. L. and Boothe, D. M. (2013) Chromosomal and plasmid-mediated fluoroquinolone resistancemechanisms among broad-spectrum-cephalosporin-resistant Escherichia coli isolates recovered from companion animals in the USA. J Antimicrob Chemother 68: 1019–1024.
  22. Van De Klundert, J. and Vliegenthart, J. (1993). PCR detection of genes coding for aminoglycoside-modifying enzymes. In: Persing et al. (editors) Diagnostic Molecular Microbiology: Principles and Applications. AMS Press, Washington, D.C., pp. 547-552.
  23. Zhang, Y.; Yang, J.; Ye, L.; Luo, Y.; Wang, W.; Zhou, W.; Cui, Z. and Han, L. (2012). Characterization of clinical multidrug-resistant Escherichia coli and Klebsiella pneumoniae isolates, 2007-2009, China. Microb Drug Resist 18: 465–470.
  24. Ziemer, C. J. and Steadham, S. R. (2003) Evaluation of the specificity of Salmonella PCR primers using various intestinal bacterial species. Letters in Applied Microbiology 37: 463-469.

Cite this Article:

International Journal of Sciences is Open Access Journal.
This article is licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) License.
Author(s) retain the copyrights of this article, though, publication rights are with Alkhaer Publications.

Search Articles

Issue June 2023

Volume 12, June 2023


Table of Contents



World-wide Delivery is FREE

Share this Issue with Friends:


Submit your Paper