RESEARCH PAPER
Antimicrobial resistant and virulence genes profiles of some Gram-negative bacteria from clinical isolates at Usmanu Danfodiyo University Teaching Hospital, Sokoto, Nigeria
More details
Hide details
1
Usmanu Danfodiyo University, Sokoto, Nigeria
2
Universiti Sains Malaysia, Malaysia
3
Ahmadu Bello University, Zaria, Nigeria
4
Ahmadu Bello University Teaching Hospital, Shika, Nigeria
5
Usmanu Danfodiyo University Teaching Hospital, Sokoto, Nigeria
Corresponding author
Tanko Nuhu
Usmanu Danfodiyo University Sokoto, Garba Nadama Road, 840212, Sokoto, Nigeria
J Pre Clin Clin Res. 2020;14(2):52-57
KEYWORDS
TOPICS
ABSTRACT
Introduction and objective:
Infections due to multidrug-resistant (MDR) Enterobacteriaceae are an ongoing global threat in their management. The aim of the study was to investigate the antiimicrobial resistance (AMR) and virulence gene profiles of MDR Gram-negative isolates in Sokoto, north-west Nigeria.
Material and methods:
A total of 578 clinical samples were collected from patients. Suspected Gram-negative bacteria were isolated from these clinical samples: vaginal swab, pus, stool, blood, wound swab and urine, using Gram-staining and conventional biochemical reactions. These isolates were further identified with an identification kit (Microgen-GN-A), and tested against a panel of 11 antibiotics. A single polymerase chain reaction (PCR) assay targeting 13 virulence gene related to adhesion (fim H, papC, and sfaS), iron chelation (iutA, and fyuA), toxins (astA, stx1, stx2, and eaeA), biofilm (bssS), and serum resistance (traT, iss, and kapsMTII) encoding genes were evaluated.
Results:
A total of 276 Gram-negative isolates were identified using the Gram stain and biochemical reactions. These organisms were further confirmed with identification kit. Of the 276 isolates, 36 organisms of interest (23 Escherichia coli, 4 Klebsiella pneumoniae and 9 Proteus mirabilis) were identified. Other Gram-negative isolates accounted for the remaining 86.9%. The majority of the isolates were resistant to cefixime (100%) and partially resistant to amikacin (19.4%).The virulence genes bssS (58.3%), fim H (44.4%), and iutA (44.4%) were the most prevalent, whereas kapsMTII (5.6%) and stx2 (2.8%) were least detected, while astA was not detected in any of the isolates.
Conclusions:
The study elucidated the prevalence of antibiotic resistance and virulence genes in Gram-negative bacteria from clinical isolates in Sokoto, north-western Nigeria. The majority of the isolates were MDR, thereby posing a public health risk.
REFERENCES (40)
1.
Srinivasan R, Karaoz U, Volegova M, Mackichan J, Kato M. Use of 16S rRNA Gene for Identification of a Broad Range of Clinically Relevant Bacterial Pathogens. PLoS One. 2015; 1–22.
2.
Iii J, Clarridge E. Impact of 16S rRNA Gene Sequence Analysis for Identification of Bacteria on Clinical Microbiology and Infectious Diseases. Clin Microbiol Rev. 2004; 17(4): 840–862.
3.
Limmathurotsakul D, Dunachie S, Fukuda K, Feasey NA, Okeke IN, Frcp AHH, et al. Personal View Improving the estimation of the global burden of antimicrobial resistant infections. Lancet Infect Dis. 2019; 3099.
http://dx.doi.org/10.1016/S147....
4.
Khalifa HO, Soliman AM, Ahmed AM, Shimamoto T, Nariya H, Matsumoto T, et al. High Prevalence of Antimicrobial Resistance in Gram-negative Bacteria Isolated from Clinical Settings in Egypt: Recalling for Judicious Use of Conventional. Microbiol Drug Resist. 2019; 00(00): 1–15.
5.
Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect. 2012; 18(3): 268–281.
http://dx.doi.org/10.1111/j.14....
6.
Chong Y, Shimoda S, Shimono N. Current epidemiology, genetic evolution and clinical impact of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae. Infection, Genetics and Evolution. 2018; 61.
7.
Munoz-Price SL, Laurent P, Bonomo RA, Schwaber MJ, Daikos GL, Cormican M, et al. Lancet Infect Dis. 2013; 13(9): 785–796.
8.
Jasovsky D, Littmann J, Zorzet A, Cars O. Antimicrobial Resistance- A Threat to the World’s Sustainable Development. Dev Dialogue Pap. 2016; 16: 1–8.
9.
Jim O. Antimicrobial Resistance: Tackling a crisis for the health and wealth of nations. Rev Antimicrob Resist. 2014; 1–20.
10.
Giraud E, Rychlik I, Cloeckaert A. Antimicrobial Resistance and Virulence Common Mechanisms. Front Cell Infect Microbiol. 2017; 8: 1–3.
12.
Chapman TA, Wu XY, Barchia I, Bettelheim KA, Driesen S, Trott D, et al. Comparison of virulence gene profiles of Escherichia coli strains isolated from healthy and diarrheic swine. Appl Environ Microbiol. 2006; 72(7): 4782–4795.
13.
Barbosa C, Nogueira S, Gadanho M, Chaves S. DNA extraction: Finding the most suitable method. Molecular Microbial Diagnostic Methods: Pathways to Implementation for the Food and Water Industries. Elsevier Inc. 2015: 135–154.
http://dx.doi.org/10.1016/B978....
14.
El-shaer S, Abdel-rhman SH, Barwa R, Hassan R. Virulence Characteristics, Serotyping and Phylogenetic Typing of Clinical and Environmental Escherichia coli Isolates. Jundishapur J Microbiol. 2018; 11(12): 1–12.
15.
Abimiku RH, Ngwai YB, Nkene IH, Bassey BE, Tsaku PA, Ibrahim T, et al. Phenotypic Detection of Extended Spectrum Beta-lactamase Resistance of Escherichia coli from Patients Attending Selected Healthcare Facilities in Nasarawa State, Nigeria. South Asian J Res Microbiol. 2019; 4(3): 1–10.
16.
Adenipekun EO, Jackson CR, Ramadan H, Iwalokun BA, Oyedeji KS, Frye JG, et al. Prevalence and multidrug resistance of Escherichia coli from community-acquired infections in Lagos, Nigeria. J Infect Dev Ctries. 2016; 10(9): 920–931.
17.
Kpoda DS, Guessennd N, Bonkoungou JI, Ouattara MB, Konan F, Ajayi A, et al. Prevalence and resistance profile of extended-spectrum beta-lactamases-producing Enterobacteriaceae in Ouagadougou, Burkina Faso. African J Microbiol Res. 2017; 11(27): 1120–1126.
18.
Dadi BR, Abebe T, Zhang L, Mihret A, Abebe W, Amogne W. Distribution of virulence genes and phylogenetics of uropathogenic Escherichia coli among urinary tract infection patients in Addis Ababa, Ethiopia. BMC Infect Dis. 2020; 20: 1–12.
19.
Eric L, Hunfeld LK, Emrich T, Haberhausen G, Wissing H, Hoeft A, et al. A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples. Med Microbiol Immunol. 2008; 197: 313–324.
20.
Hallin M, Maes N, Byl B, Jacobs F, Gheldre Y De, Struelens MJ. Clinical Impact of a PCR Assay for Identification of Staphylococcus aureus and Determination of Methicillin Resistance Directly from Blood Cultures. J Clin Microbiol. 2003; 41(8): 3942–3944.
21.
Barbut F, Monot M, Rousseau A, Cavelot S, Simon T, Burghoffer B, et al. Rapid diagnosis of Clostridium difficile infection by multiplex real-time PCR. Eur J Clin Microbiol Infect Dis. 2011; 30: 1279–1285.
22.
Beceiro A, Tomás M, Bou G. Antimicrobial resistance and virulence: A successful or deleterious association in the bacterial world? Clin Microbiol Rev. 2013; 26(2): 185–230.
23.
Ayukekbong JA, Ntemgwa M, Atabe AN. The threat of antimicrobial resistance in developing countries: causes and control strategies. Antimicrob Resist Infect Control. 2017; 6(47): 1–8.
24.
Ouedraogo AS, Sanou M, Kissou A, Sanou S, Solaré H, Kaboré F, et al. High prevalence of extended-spectrum beta-lactamase producing Enterobacteriaceae among clinical isolates in Burkina Faso. BMC Infect Dis. 2016; 16(1): 1–9.
http://dx.doi.org/10.1186/s128....
25.
Sáez-lópez E, Cossa A, Benmessaoud R, Madrid L. Characterization of Vaginal Escherichia coli Isolated from Pregnant Women in Two Different African Sites. PLoS One. 2016; 1–10.
26.
Mshana SE, Matee M, Rweyemamu M. Antimicrobial resistance in human and animal pathogens in Zambia, Democratic Republic of Congo, Mozambique and Tanzania: an urgent need of a sustainable surveillance system. Ann Clin Microbiol Antimicrob. 2013; 12(1): 1.
27.
Ugwu MC, Igbokwe JO, Okezie U, Eze PM, Ejikeugwu CP, Esimone CO. Prevalence of ESBLs and MBLs among Escherichia coli and Klebsiella pneumoniae isolates from a Nigerian Abattoir. J Trop Dis. 2018; 6(2): 2–6.
28.
Giwa FJ, Ige OT, Haruna DM, Yaqub Y, Lamido TZ US. Extended--Spectrum Beta-lactamase Production and Antimicrobial Susceptibility Pattern of Uropathogens in a Tertiary Hospital in Northwestern Nigeria. Ann Trop Pathol. 2018; 9: 11–16.
29.
Boroumand M, Sharifi A, Manzouri L, Khoramrooz SS, Khosravani A. Evaluation of pap and sfa Genes Relative Frequency P and S Fimbriae Encoding of Uropathogenic Escherichia coli Isolated from Hospitals and Medical Laboratories; Yasuj City, Southwest Iran. Iran Red Crescent Med J. 2019; 21(8): 1–8.
30.
Ramirez MS, Tolmasky ME. Amikacin: Uses, Resistance, and Prospects for Inhibition. Molecules. 2017; 22(1957).
31.
Gharrah MM, Mostafa El-Mahdy A, Barwa RF. Association between Virulence Factors and Extended Spectrum Beta-Lactamase Producing Klebsiella pneumoniae Compared to Non-producing Isolates. Interdiscip Perspect Infect Dis. 2017; 2017(27): 251–264.
32.
Ghasemian A, Mobarez AM, Peerayeh SN, Abadi ATB. The association of surface adhesin genes and the biofilm formation among Klebsiella oxytoca clinical isolates. New Microbes New Infect. 2019; 27: 36–39.
https://doi.org/10.1016/j.nmni....
33.
Bellifa S, Hassaine H, Balestrino D, Charbonnel N, Imane M, Terki IK, et al. Evaluation of biofilm formation of Klebsiella pneumoniae isolated from medical devices at the University Hospital of Tlemcen, Algeria. African J Microbiol Res. 2013; 7(49): 5558–5564.
34.
Sarkar S, Vagenas D, Schembri MA, Totsika M. Biofilm formation by multidrug resistant Escherichia coli ST131 is dependent on type 1 fimbriae and assay conditions. Pathog Dis. 2016; 74: 1–5.
35.
Sarowska J, Koloch BF, Kmiecik AJ, Madrzak MF, Ksiazczyk M, Ploskonska GB, et al. Virulence factors, prevalence and potential transmission of extraintestinal pathogenic Escherichia coli isolated from different sources: recent reports. Gut Pathog. 2019; 11(10): 1–16.
https://doi.org/10.1186/s13099....
36.
Rahdar M, Rashki A, Miri HR, Ghalehnoo MR. Uropathogenic Escherichia coli Isolates Collected From Patients With Urinary Tract Infection. Jundishapur J Microbiol. 2015; 8(8): 1–6.
37.
Düzgün AÖ, Okumuş F, Saral A. Determination of antibiotic resistance genes and virulence factors in Escherichia coli isolated from Turkish patients with urinary tract infection. J Brazilian Soc Trop Med. 2019; 52: 1–5.
38.
Wilson BR, Bogdan AR, Miyazawa M, Hashimoto K. Siderophores in Iron Metabolism: From Mechanism to Therapy Potential. Trends Mol Med. 2017; 22(12): 1077–1090.
39.
Abdelmegeed ES, Barwa R, Abd KH, Galil E. Comparative study on prevalence and association of some virulence factors with extended spectrum beta-lactamases and AmpC producing Escherichia coli. African J Microbiol Res. 2015; 9(17): 1165–74.
40.
Hassan R, El-Naggar W, El-Sawy E, E-MA. Characterization of Some Virulence Factors Associated with Enterbacteriaceae Isolated From Urinary Tract Infections in Mansoura Hospitals. Egypt J Med Microbiol. 2011; 20(2): 9–17.