Cross-Sectional Study of Extended Spectrum β-Lactamase Genes (SHV, TEM, CTX-M and OXA) in Klebsiella Species from Clinical Specimens, in Port Harcourt, Nigeria

Ezinna Isu Okogeri¹, Smart Enoch Amala^1, , Easter Godwin Nwokah¹ and Tombari Pius Monsi¹

¹Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria

Cite this paper:
Ezinna Isu Okogeri, Smart Enoch Amala, Easter Godwin Nwokah and Tombari Pius Monsi. Cross-Sectional Study of Extended Spectrum β-Lactamase Genes (SHV, TEM, CTX-M and OXA) in Klebsiella Species from Clinical Specimens, in Port Harcourt, Nigeria. American Journal of Medical Sciences and Medicine. 2020; 8(5):180-186. doi: 10.12691/ajmsm-8-5-3

Abstract

Infections caused by Klebsiella sp. are increasingly becoming difficult to treat partly due to the rise in trend of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella sp. This study aimed to detect the prevalence of ESBL-producing Klebsiella sp. and SHV, TEM, CTX-M, and OXA beta-lactamase encoding genes in isolates of Klebsiella sp. from two healthcare facilities in Port Harcourt. A cross-sectional study of 146 clinical specimens was analyzed using standard bacteriological identification techniques. ESBL-producing Klebsiella sp. was identified using clinical laboratory standard institute standards. Beta-lactamase genes SHV, TEM, OXA, and CTXM were extracted and amplified. Of the 146 clinical specimens; 47 (32.2%) were from males and 99 (67.8%) females. The number of Klebsiella sp. isolated from the female samples 11 (73.3%) was more than 2-fold higher than those isolated from male samples 4 (26.7%). Eighty different bacterial isolates were obtained from which 15 (18.8%) were Klebsiella sp. while 12 (80%) of the identified Klebsiella sp. were β-lactamase producing Klebsiella sp. The prevalence of the Klebsiella sp. according to the different samples were 8 (53.3%), 1 (6.7%), 2 (13.3%), 2 (13.3%), 1 (6.7%), 1 (6.7%) were obtained from urine, endocervical, sputum, high vaginal, throat, hospital environment samples respectively. Isolates 2, 4-6, and 9-11 were positive for SHV gene (293 bp); all the Klebsiella isolates were positive for TEM gene (840 bp); isolates 4-8 showed positive bands for CTX-M gene (550 bp); and isolates 2, 3, 6, 8, and 12 showed positive bands for OXA gene (908 bp). The urine sample numbered 106, 42, and 55 exhibited complete resistance to all the antibiotics used and showed different types of ESBL genes: SHV; SHV, TEM, CTX-M and OXA; and OXA, and TEM respectively. This study shows that β-lactamase genes were differentially expressed in the various types of samples collected. Again, total resistance to beta-lactam drugs may not be completely dependent on the constitutive expression of several ESBL genes but TEM is present in all the sample that showed complete resistance.

Keywords:
beta-lactamase gene antimicrobial resistance Klebsiella extended spectrum beta-lactamase

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Woerther, P.L., Burdet, C., Chachaty, E. and Andremont, A. “Trends in human fecal carriage of extended-spectrum beta-lactamases in the community: Toward the globalization of CTX-M,” Clin Microbiol Rev.., 24 (4), 744-758, October 2013.
[2]	Karanika, S., Karantanos, T., Arvanitis, M., Grigoras, C. and Mylonakis, E. “Fecal Colonization with Extendedspectrum Beta-lactamase-Producing Enterobacteriaceae and Risk Factors among Healthy Individuals: A Systematic Review and Metaanalysis,” Clin Infect Dis., 63 (3), 310-318, August 2016.
[3]	European Centre for Disease Prevention and Control (ECDC). Antimicrobial resistance surveillance in Europe 2014. Annual Report of the European Antimicrobial Resistance Surveillance Network (EARSNet). Stockholm: ECDC; 2015.
[4]	Tumbarello, M., Spanu, T., Di Bidino, R., Marchetti, M., Ruggeri, M. and Trecarichi, E.M. “Costs of bloodstream infections caused by Escherichia coli and influence of extended-spectrum-beta-lactamase production and inadequate initial antibiotic therapy,” Antimicrob Agents Chemother., 54 (10), 4085-4091, October 2010.
[5]	Huizinga, P., van den Bergh, M.K., van Rijen, M., Willemsen, I., van Veer, N., and Kluytmans, J. “Proton Pump Inhibitor Use Is Associated With Extended-Spectrum β-Lactamase-Producing Enterobacteriaceae Rectal Carriage at Hospital Admission: A Cross-Sectional Study,” Clin. Infect Dis. 64 (3), 361-363, February 2016.
[6]	Reuland, E.A., Naiemi, N., Kaiser, A.M., Heck, M., Kluytmans. J.A.J.W. and Savelkoul, P.H.M. “Prevalence and risk factors for carriage of ESBL-producing Enterobacteriaceae in Amsterdam,” J Antimicrob Chemother., 71, 1-7, April, 2016.
[7]	Shitrit, P., Reisfeld, S., Paitan, Y., Gottesman, B.S., Katzir, M., Paul, M. and Chowers, M. “Extended-spectrum beta-lactamase-producing Enterobacteriaceae carriage upon hospital admission: Prevalence and risk factors,” J Hosp Infect., 85, 230-232, November, 2013.
[8]	Huizinga, P., Schrauwen, E., García-Cobos, S., Willemsen, I., Verhulst, C., Friedrich, A.W., Savelkoul, P.H.M., Rossen, J.W. and Kluytmans, J. “Extended-spectrum beta-lactamase producing Enterobacteriaceae (ESBL-E) isolated from bean sprouts in the Netherlands,” PLoS One, 13 (8), e0203338, August, 2018.
[9]	Arcilla, M.S., Hattem, J.M. van Haverkate, M.R., Bootsma, M.C.J., Genderen, P.J.J. and van Goorhui,s A. “Import and spread of extended-spectrum β-lactamase-producing Enterobacteriaceae by international travellers (COMBAT study): a prospective, multicentre cohort study,” Lancet Infect Dis. 17 (1), 78-85, January, 2017.
[10]	Barreto, M.I.D.M., Ignatius, R.P.D.M., Pfuller, R.D.M., Friedrich-Janicke, B.D.M., Steiner, F.D.M. and Paland, M.D.M. “High carriage rate of ESBL-producing Enterobacteriaceae at presentation and follow-up among travellers with gastrointestinal complaints returning from India and Southeast Asia,” J Travel Med., 23 (2), 1-7, February, 2016.
[11]	Valverde, A., Grill, F., Coque, T.M., Pintado, V., Baquero, F., Cantón, R., Cobo, J. “High rate of intestinal colonization with extended-spectrum-β- lactamase-producing organisms in household contacts of infected community patients,” J Clin Microbiol,46 (8), 2796-2799, August 2008.
[12]	Meyer E, Gastmeier P, Kola A, Schwab F. “Pet animals and foreign travel are risk factors for colonization with extended-spectrum beta-lactamase-producing Escherichia coli,” Infection. 40 (6), 685-687, December 2012.
[13]	Kim, J., Ding, T., and Ahn, J. “Relationship between β-lactamase production and resistance phenotype in Klebsiella pneumoniae strains,” FEMS Microbiology Letters, 364 (13), July, 2017.
[14]	Doosti A, Pourabbas M, Arshi A et al. “TEM and SHV genes in Klebsiella pneumoniae isolated from cockroaches and their antimicrobial resistance pattern,” Osong Pub Health Res Perspect, 6 (1), 3-8. December 2015.
[15]	Knothe H, Shah P, Krcmery V, Antal M, Mitsuhashi S. “Transferable resistance to cefotaxime, cefoxitin, cefamandole and cefuroxime in clinical isolates of Klebsiella pneumoniae and Serratia marcescens,” Infection. 11 (6), 315-317, December 1983.
[16]	Peleg, A.Y., Hooper, D.C. “Hospital-acquired infections due to gram-negative bacteria,” N Engl J Med., 362 (19), 1804-1813, May 2010.
[17]	Philippon, A., Arlet, G., andLagrange, P.H. “Origin and Impact of plasmid-mediated extended spectrum beta-lactamases,” European Journal of Clinical Microbiology Infectious Diseases, 13 (1), 17-29, 1994.
[18]	Barry, G.H., and Barlow, M. “Revised Ambler classification of b-lactamases,” J Antimicrob Chemother, 55 (6), 1050-1051, June, 2005; 55.
[19]	Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing; twenty-second informational supplement. CLSI document M02-A12. Clinical and Laboratory Standards Institute, Wayne, PA, 2012
[20]	Cao, X., Xu, X., Cheng, J. and Zhang, K. “Molecular Characterization of clinical multidrug-resistant Klebsiella pneumoniae isolates,” Ann. Clin. Microb. Anti., 13, 16, May, 2014.
[21]	Sundin, D.R. “Hidden Beta-Lactamases in the Enterobacteriaceae- Dropping the Extra Disks for Detection, Part II,” Clin Microbiol Newsletter, 31, (7), 47-52, April 2009.
[22]	Tavakoly, T., Jamali, S., Mojtahedi, A., Khan Mirzaei, M., Shenagari, M. “The prevalence of CMY-2, OXA-48 and KPC-2 genes in clinical isolates of Klebsiella spp.,” Cell Mol Biol., 64 (3), 40-44, February 2018.
[23]	Monsi, T.P., Abbey, S.D., Wachukwu, C.K. and Wokem, G.N. “Plasmid-mediated resistance in extended spectrum beta-lactamase-producing Klebsiella pneumoniae isolates at rivers state university teaching hospital,” European Journal of Pharmaceutical and Medical Research, 6 (2), 75-82, January 2019.
[24]	Monsi, T.P., Abbey, S.D., Wachukwu, C.K. and Wokem, G.N. “Growth and resistance phenotypes of Klebsiella pneumoniae pre-treated with herbal drugs,” European Journal of Pharmaceutical and Medical Research, 6 (2), 65-74, January, 2019.
[25]	Villegas, M.V., Blanco, M.G., Sifuentes-Osornio, J. and Rossi, F. “Increasing prevalence of extended-spectrum-beta-lactamase among Gramnegative bacilli in Latin America--2008 update from the Study for Monitoring Antimicrobial Resistance Trends (SMART),” Braz J Infect Dis. 15 (1),34-39, January, 2011.
[26]	Schito, G.C., Naber, K.G., Botto, H., Palou, J., Mazzei, T.,and Gualco, L. “The ARESC study: an international survey on the antimicrobial resistance of pathogens involved in uncomplicated urinary tract infections,” Int J Antimicrob Agents, 34 (5), 407-413, November, 2009.
[27]	Reid, C.B., Steele, L., Pasquill, K., Parfitt, E.C. amd Laupland, K.B. “Occurrence and determinants of Klebsiella species bloodstream infection in the western interior of British Columbia, Canada,” BMC Infect Dis. 19, 1070, December, 2019.
[28]	Al-Hasan, M.N., Lahr, B.D., Eckel-Passow, J.E., and Baddour, L.M. “Epidemiology and outcome of Klebsiella species bloodstream infection: a population-based study,” Mayo Clin Proc. 85 (2), 139-44, February, 2010.
[29]	Davis, T.J., and Matsen, J.M. “Prevalence and characteristics of Klebsiella species: relation to association with a hospital environment,” J Infect Dis, 130 (4), 402-405, October, 1974.
[30]	Rosenthal, S., and Tager, I.B. Prevalence of gram-negative rods in the normal pharyngeal flora,” Ann Intern Med, 83 (3), 355-357, September, 1975.
[31]	Rose, H.D. and Schreier, J. “The effect of hospitalization and antibiotic therapy on the gram-negative fecal flora,” Am J Med Sci. 255, 228-236, April, 1968.
[32]	Biradar, S. and Roopa, C. “Isolation and Antibiogram of Klebsiella Species from Various Clinical Specimens,” International Journal of Current Microbiology and Applied Sciences, 4 (9), 991-995, 2015.
[33]	Demirdag, K. and Hosoglu, S. “Epidemiology and risk factors for ESBL-producing Klebsiella pneumoniae: a case control study,” J Infect Dev Ctries, 4 (7), 717-722, November, 2010.
[34]	Vasaikar, S., Obi, L., Morobe, I. and Bisi-Johnson, M. “Molecular Characteristics and Antibiotic Resistance Profiles of Klebsiella Isolates in Mthatha, Eastern Cape Province, South Africa,” International Journal of Microbiology, 2017, 8486742, January, 2017.
[35]	Marr, A.K., J. Overhage, M. Bains, and Hancock, R.E. “The Lon protease of Pseudomonas aeruginosa is induced by aminoglycosides and is involved in biofilm formation and motility,” Microbiology, 153 (2), 474-482, February, 2007.
[36]	Brazas, M.D., Breidenstein, E.B., Overhage, J., and Hancock, R.E. “Role of lon, an ATP-dependent protease homolog, in resistance of Pseudomonas aeruginosa to ciprofloxacin,” Antimicrob Agents Chemother, 51 (12), 4276-4283, December, 2007.
[37]	Li, J., Xie, S., Ahmed, S., Wang, F., Gu, Y., Zhang, C., Chai, X., Wu, Y., Cai, J. and Cheng, G. “Antimicrobial Activity and Resistance: Influencing Factors,” Front Pharmacol, 8, (364), 13, June, 2017.