Abstract
The Gram-negative bacteria were isolated from various samples and antibiotic susceptibility testing were performed. The strains were screened by PCR for class 1 integron, virulence and antibiotic resistance genes. blaOXA-23 and blaOXA-51 genes positive samples were cloned into the pGEM-T vector and the plasmids were sent for DNA sequence analysis. The isolates were showed resistance to quinolone, cephalosporin, aminoglycoside, carbapenem and penicillin group antibiotics. Virulence factor genes were identified in 121 strain. The most common virulence gene was determined to be aer (29.78%), followed by sfa (28.19%), afa (27.12%), cnf (21.27%), pap (12.76%) and hly (12.76%). In addition, 34 different virulence patterns were observed. Plasmid-mediated quinolone resistance genes were detected 15 of67 quinolone-resistant isolates. Among carbapenem resistant strains, blaOXA-23 gene was detected in 16 and blaOXA-51 gene in 19 isolates. The strains producing OXA-23 and OXA-51 were Klebsiella oxytoca, Acinetobacter baumannii, E.coli and P.mirabilis. In this study, clinical isolate K. oxytoca carrying both blaOXA-23 and blaOXA-51 genes were identified first time from Türkiye. A high predominance (88.82%) of class 1 integron was detected. In addition, determined that 46.27% of the isolates had the ability to form biofilms. In conclusion since the limited treatment options of antibiotic-resistant Gram-negative pathogens that cause serious infections are an important problem in terms of public health it is important to investigate antibiotic resistance, virulence factor genes and biofilm formation in these pathogens.
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Abbreviations
- K.oxytoca :
-
Klebsiella oxytoca
- KPC:
-
Klebsiella pneumoniae carbapenemase
- IMP:
-
Imipenem-resistant Pseudomonas
- VIM:
-
Verona integron-encoded Metallo beta lactamase
- OXA:
-
Oxacillinases
- LB:
-
Luria Broth
- NDM:
-
New Delhi Metallo beta lactamase
- PCR:
-
Polymerase chain reaction
- E. coli :
-
Escherichia coli
- aer:
-
Aerobactin
- cnf 1:
-
Necrotizing factor 1
- pap:
-
P fimbriae
- sfa:
-
S fimbriae
- afaI:
-
Afimbrial adhesin I
- hly:
-
Hemolysin
- PMQR:
-
Plasmid-mediated quinolone resistance genes
References
Ahmed HA, El-Hofy FI, Shafik SM, Abdelrahman MA, Elsaid GA (2016) Characterization of virulence-associated genes, antimicrobial resistance genes, and class 1 Integrons in salmonella enterica serovar typhimurium isolates from chicken meat and humans in Egypt. Foodborne Pathog Dis 13(6):281–288. https://doi.org/10.1089/fpd.2015.2097
Akyıldız E, Saral A, Köse T, Özad Düzgün A (2020) Investigation of virulence factor genes and extended Spectrum Beta lactamases in Clinical E.coli isolates. Güfbed/Gustij 10(1):122–127. https://doi.org/10.17714/gumusfenbil.571958
Babiker A, Clarke LG, Saul M, Gealey JA, Clancy CJ, Nguyen MH, Shields RK (2021) Changing epidemiology and decreased mortality associated with Carbapenem-resistant Gram-negative bacteria, 2000–2017. 73:e4521–e4530. https://doi.org/10.1093/cid/ciaa1464
Breijyeh Z, Jubeh B, Karaman R (2020) Resistance of gram-negative Bacteria to current antibacterial agents and approaches to resolve it. Molecules 25(6):1340. https://doi.org/10.3390/molecules25061340
Brink AJ (2019) Epidemiology of carbapenem-resistant gram-negative infections globally. Curr Opin Infect Dis 32:609–616. https://doi.org/10.1097/QCO.0000000000000608
Brown S, Young HK, Amyes SGB (2005) Characterisation of OXA-51, a novel class D carbapenemase found in genetically unrelated clinical strains of Acinetobacter baumannii from Argentina. Clin Microbiol Infect 11:15–23. https://doi.org/10.1111/j.1469-0691.2004.01016.x
Budak S, Aktaş Z, Oncul O, Acar A, Ozyurt M, Turhan V Gorenek L (2013) Detection of OXA-51 Carbapenemase gene in Klebsiella pneumoniae: a case report and a new dimension on Carbapenemase resistance. J Mol Genet Med 7:2. https://doi.org/10.4172/1747-0862.1000063
Carattoli A (2009) Resistance plasmid families in Enterobacteriaceae. Chemotherapy 53(6):2227–2238. https://doi.org/10.1128/AAC.01707-08
Cepas V, Soto SM (2020) Relationship between virulence and resistance among gram-negative Bacteria. Antibiotics (Basel) 209(10):719. https://doi.org/10.3390/antibiotics9100719
Cestari SE, Ludovico MS, Martins FH, da Rocha SPD, Elias WP, Pelayo JS (2013) Molecular detection of HpmA and HlyA hemolysin of uropathogenic Proteus mirabilis. Curr Microbiol 67(6):703–707. https://doi.org/10.1007/s00284-013-0423-5
Chen S, Hu F, Liu Y, Zhu D, Wang H, Zhang Y (2011) Detection and spread of carbapenem-resistant Citrobacter freundii in a teaching hospital in China. Am J Infect Control 39:e55–e60. https://doi.org/10.1016/j.ajic.2011.02.009
Çimen M, Düzgün Özad A (2021) Antibiotic induced biofilm formation of novel multidrug resistant Acinetobacter baumannii ST2121 clone. Acta Microbiol Immunol Hung 68(2):80–86. https://doi.org/10.1556/030.2020.01240
Davoudi-Monfared E, Khalili H (2018) The threat of carbapenem-resistant gram-negative bacteria in a Middle East region. Infect Drug Resist 11:1831–1880. https://doi.org/10.2147/IDR.S176049
Donnenberg M, Welch RA (1995) Virulence determinants of uropathogenic Escherichia coli. In: Mobely HLT, Warren JW (eds) Urinary tract infections. Molecular pathogenesis and clinical management. American Society for Microbiology, Washington, DC, pp 135–174
Düzgün AÖ, Okumuş F, Saral A, Çiçek AÇ, Cinemre S (2019) Determination of antibiotic resistance genes and virulence factors in Escherichia coli isolated from Turkish patients with urinary tract infection. Rev Soc Bras Med Trop 52:e20180499. https://doi.org/10.1590/0037-8682-0499-2018
Eftekhar F, Altayar F, Khidaei H (2018) Plasmid-mediated class 1 and 2 Integron carriage in drug-resistant nosocomial isolates of Acinetobacter baumannii. Arch Clin Infect Dis 13(1). https://doi.org/10.5812/archcid.57813
Glasner C, Albiger B, Buist G, Andrašević AT, Cantón R, Carmeli Y, Friedrich AW, Giske CG, Glupczynski Y, Gniadkowski M et al (2013) Carbapenemase-producing Enterobacteriaceae in Europe: a survey among national experts from 39 countries. Eurosurveillance. 18:20525. https://doi.org/10.2807/1560-7917
Heritier C, Poirel L, Fournier PE, Claverie JM, Raoult D, Nordmann P (2005) Characterization of the naturally occurring oxacillinase of Acinetobacter baumannii. Antimicrob Agents Chemother 49:4174–4179. https://doi.org/10.1128/AAC.49.10.4174-4179.2005
Hussein EI, Al-Batayneh K, Masadeh MM, Dahadhah FW, Al Zoubi MS, Aljabali AA, Alzoubi KH (2020) Assessment of pathogenic potential, virulent genes profile, and antibiotic susceptibility of Proteus mirabilis from urinary tract infection. Int J Microbiol 2020:1231807. https://doi.org/10.1155/2020/1231807
Iraz M, Özad Düzgün A, Sandallı C, Doymaz MZ, Akkoyunlu Y, Saral A, Peleg AY, Özgümüş OB, Beriş FŞ, Karaoğlu H, Çopur Çiçek A (2015) Distribution of β-lactamase genes among carbapenem-resistant Klebsiella pneumoniae strains isolated from patients in Turkey. Ann Lab Med 35:595–601. https://doi.org/10.3343/alm.2015.35.6.595
Iseppi R, Di Cerbo A, Messi P, Sabia C (2020) Antibiotic resistance and virulence traits in vancomycin-resistant enterococci (VRE) and extended-Spectrum -lactamase/AmpC-producing (ESBL/AmpC) Enterobacteriaceae from humans and pets. Antibiotics 9:152. https://doi.org/10.3390/antibiotics9040152
Jeong HW, Cheong HJ, Kim WJ, Kim MJ, Song KJ, Song JW, Kim HS, Roh KH (2009) Loss of the 29-kilodalton outer membrane protein in the presence of OXA-51-like enzymes in Acinetobacter baumannii is associated with decreased imipenem susceptibility. Microb Drug Resist 15:151–158. https://doi.org/10.1089/mdr.2009.0828
Johnson JR (1991) Virulence factors in Escherichia coli urinary tract infection. Clin Microbiol Rev 4(1):80–128. https://doi.org/10.1128/CMR.4.1.80
Kareem SM, Al-Kadmy IMS, Kazaal SS, Ali ANM, Aziz SN, Makharita RR, Algammal AM, Al-Rejaie S, Behl T, Batiha GES et al (2021) Detection of gyrA and parC mutations and prevalence of plasmid-mediated quinolone resistance genes in Klebsiella pneumonia. Infect Drug Resist. 12(14):555–563. https://doi.org/10.2147/IDR.S275852
Kargar M, Mohammadalipour Z, Doosti A, Lorzadeh S, Japoni-Nejad A (2014) High prevalence of class 1 to 3 Integrons among multidrug-resistant Diarrheagenic Escherichia coli in southwest of Iran. Osong Public Health Res Perspect 5(4):193–198. https://doi.org/10.1016/j.phrp.2014.06.003
Lee HW, Koh YM, Kim J, Lee JC, Lee YC, Seol SY, Cho DT, Kim J (2008) Capacity of multidrugresistant clinical isolates of Acinetobacter baumannii to form biofilm and adhere to epithelial cell surfaces. Clin Microbiol Infect 14:49–54. https://doi.org/10.1111/j.1469-0691.2007.01842.x
Lin MF, Lan CY (2014) Antimicrobial resistance in Acinetobacter baumannii: from bench to bedside. World J Clin Cases 2:787–814. https://doi.org/10.12998/wjcc.v2.i12.787
Manohar P, Leptihn S, Lopes BS, Nachimuthu R (2021) Dissemination of carbapenem resistance and plasmids encoding carbapenemases in gram-negative bacteria isolated in India. JAC Antimicrob Resist 3(1):dlab015. https://doi.org/10.1093/jacamr/dlab015
Martínez-Martínez L, Pascual A, Jacoby GA (1998) Quinolone resistance from a transferable plasmid. Lancet. 351(9105):797–799. https://doi.org/10.1016/S0140-6736(97)07322-4
Navia MM, Ruiz J, Vila J (2002) Characterization of an integron carrying a new class D beta-lactamase (OXA-37) in Acinetobacter baumannii. Microb Drug Resist 8:261–265. https://doi.org/10.1089/10766290260469516
Oliveira J, Reygaert WC (2019) Gram negative Bacteria. StatPearls Publishing, Treasure Island
Perez F, Hujer AM, Hujer KM, Decker BK, Rather PN, Bonomo RA (2007) Global challenge of multidrug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 51:3471–3484. https://doi.org/10.1128/AAC.01464-06
Poirel L, Naas T, Nordmann P (2010) Diversity, epidemiology, and genetics of class D beta-lactamases. Antimicrob Agents Chemother 54:24–38. https://doi.org/10.1128/AAC.01512-08
Ruiz J (2003) Mechanisms of resistance to quinolones: target alterations, decreased accumulation and DNA gyrase protection. J Antimicrob Chemother 51(5):1109–1117. https://doi.org/10.1093/jac/dkg222
Ruppé E, Woerther PL, Barbier F (2015) Mechanisms of antimicrobial resistance in gram-negative bacilli. Ann Intensiv Care 5:61. https://doi.org/10.1186/s13613-015-0061-0
Sabbagh P, Rajabnia M, Maali A, Ferdosi-Shahandashti E (2021) Integron and its role in antimicrobial resistance: a literature review on some bacterial pathogens. Iran J Basic Med Sci 24(2):136–142. https://doi.org/10.22038/ijbms.2020.48905.11208
Schroeder M, Brooks BD, Brooks AE (2017) The complex relationship between virulence and antibiotic resistance. Genes. 8:39. https://doi.org/10.3390/genes8010039
Tajbakhsh E, Ahmadi P, Abedpour-Dehkordi E, Arbab-Soleimani N, Khamesipour F (2016) Biofilm formation, antimicrobial susceptibility, serogroups and virulence genes of uropathogenic E. coli isolated from clinical samples in Iran. Antimicrob resist. Infect Control 1(5):11. https://doi.org/10.1186/s13756-016-0109-4
Zhang D, Xia J, Xu Y, Gong M, Zhou Y, Xie L et al (2016) Biological features of biofilm-forming ability of Acinetobacter baumannii strains derived from 121 elderly patients with hospital-acquired pneumonia. Clin Exp Med 16:73–80. https://doi.org/10.1007/s10238-014-0333-2
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All experiments were performed by GY and AÖD. AÖD wrote the paper. Authors reviewed the manuscript.
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Ethical approval was obtained from the ethics committee of Gumushane University Ethics committee date: 29.12.2021 and number: 2021/8.
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Özad Düzgün, A., Yüksel, G. Detection of virulence factor genes, antibiotic resistance genes and biofilm formation in clinical Gram-negative bacteria and first report from Türkiye of K.oxytoca carrying both blaOXA-23 and blaOXA-51 genes. Biologia 78, 2245–2251 (2023). https://doi.org/10.1007/s11756-023-01355-0
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DOI: https://doi.org/10.1007/s11756-023-01355-0