Molecular characterization of Pseudomonas aeruginosa isolates from various clinical specimens in Khartoum/Sudan: Antimicrobial resistance and virulence genes
Keywords:Pseudomonas aeruginosa, Antimicrobial resistance, Virulence factors, Khartoum/Sudan
Background: Pseudomonas aeruginosa is a pathogenic organism responsible for frequent wound and nosocomial infections worldwide. Its infections are difficult to control since the organism is known to rapidly develop antibiotic resistance and becomes multidrug-resistant (MDR) during treatment of patients.
Aim of the study: This study was intended to investigate the occurrence of certain important types of (ESBL) and (MBL) enzymes in association with important specific virulence factors associated with P. aeruginosa clinical isolates from Khartoum, Sudan.
Methods: This study investigated 70 P. aeruginosa isolates which were collected from patients admitted to four major hospitals in Khartoum (Fedail, Ribat, Ibn Sina and Soba hospitals). These isolates were recovered from 40 wound swabs (57.1%), 27 urine samples (38.6%), and 3 pleural fluid samples (4.3%) of patients. Higher numbers of isolates were recovered from males 42 (60%) than in females 28 (40%). All P. aeruginosa isolates were first confirmed by conventional biochemical and second using molecular PCR tests. PCR methods were also used for detecting the presence of the virulence genes ToxA, AlgD, LasB, exoS, exoU, CTX, GES-1, and genes of VIM, IMP, KPC, CTX, VEB-1 and SHV-1.
Results: Antimicrobial susceptibility testing of P. aeruginosa isolates showed a high resistance to azetronam 49 (70%), followed by ceftazidime 32 (45.7%), 16 ciprofloxacin (22.9%), gentamicin 13 (18.6 %), piperacillin-tazobactam 11 (15.7%), amikacin 9 (12.9 %), and imipenem 6 (8.6%) showed the least resistance. All isolates were positive for algD and lasB (100%), followed by toxA (90%), exoS (34.3), exoU (24.3%), respectively. The rates of detected ESBL genes blaTEM, blaCTX-m, blaSHV-1,GES-1, were 3.3%, 6.6%, 10%, 3.3%,10%, respectively, but all isolates were negative for bla-KPC and bla- VIM and IMP . The percentages of pigment production were 61.4% for pyocyanin, 37.1% for pyoverdin and 1.4% for pyorubin.
Conclusion: The study demonstrated high rates of antimicrobial resistance markers to most commonly used antibiotics in treatment of P. aeruginosa infections. The majority of the isolates from urine and wound samples carried at least three potential virulence factor genes of algD, lasB and toxA and without any significant relation to their antimicrobial resistance markers.
Shehabi AA, Haider AA, Fayyad MK., Frequency of antimicrobial resistance markers among Pseudomonas aeruginosa and Escherichia coli isolates from municipal sewage effluent water and patients in Jordan. IAJA 2011; 1(1-8).
Al Dawodeyah HY, Obeidat N, Abu-Qatouseh LF, Shehab, AA, Antimicrobial resistance and putative virulence genes of Pseudomonas aeruginosa isolates from patients with respiratory tract infection. Germs 2018; 8(1), p.31.
Paling, F.P., Wolkewitz, M., Depuydt, P., De Bus, L., Sifakis, F., Bonten, M.J. et al. 2017. P. aeruginosa colonization at ICU admission as a risk factor for developing P. aeruginosa ICU pneumonia. Antimicrob Resist Infect Control 2017; 6(1), p.38.
Harris AD, Jackson SS, Robinson G, Pineles L Leekha S Thom KA, et al. Pseudomonas aeruginosa colonization in the intensive care unit: prevalence, risk factors, and clinical outcomes. Infect Cont Hosp EP 2016; 37(5), pp.544-548.
Driscoll JA, Brody, SL, Kollef M.H. The epidemiology, pathogenesis and treatment of Pseudomonas aeruginosa infections. Drugs 2007; 67(3), pp.351-368.
Shehabi, A.A., Masoud, H. and Balkam Maslamani, F.A., 2005. Common antimicrobial resistance patterns, biotypes and serotypes found among Pseudomonas aeruginosa isolates from patient's stools and drinking water sources in Jordan. J Chemother 2005; 17(2), pp.179-183.
LaBauve, A.E. and Wargo, M.J. Growth and laboratory maintenance of Pseudomonas aeruginosa. Curr Protoc Microbiol. 2012; 25(1),6E-1.
Persson, A., Study of Pseudomonas aeruginosa and different wound dressing products. Master of Science Thesis, CHALMERS UNIVERSITY OF TECHNOLOGY SE â€“ 412 96 GÃ¶teborg Sweden, 2010.
Mahmoud, A.B., Zahran, W.A., Hindawi, G.R., Labib, A.Z. and Galal, R., 2013. Prevalence of multidrug-resistant Pseudomonas aeruginosa in patients with nosocomial infections at a university hospital in Egypt, with special reference to typing methods.
J Virol Microbiol, 13, pp.165-59.
Rumbaugh KP, Griswold , JA, Hamood AN. Pseudomonas aeruginosa strains obtained from patients with tracheal, urinary tract and wound infection: variations in virulence factors and virulence genes. J Hosp Infect 1999; 43(3), 211-218
Toder DS, Ferrell SJ, Nezezon JL, Rust L. Iglewski BH. lasA and lasB genes of Pseudomonas aeruginosa: analysis of transcription and gene product activity. Infect Immun 1994; 62(4), 1320-1327.
Deretic V, Gill JF, Chakrabarty AM. Gene algD coding for GDPmannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa. J bacteriol 1987; 169(1), 351-358.
Crousilles A, Maunders E, Bartlet S, Fan C, Ukor EF , Abdelhamid Y, et al. Which microbial factors really are important in Pseudomonas aeruginosa infections?. Fut Microb 2015; 10(11),1825-1836.
Tam VH, Schilling AN, LaRocco MT, Gentry LO, Lolans K, Quinn JP. Prevalence of AmpC overâ€expression in bloodstream isolates of Pseudomonas aeruginosa. Clin Microbiol Infect 2007; 13(4), 413-418.
Bonomo RA, Szabo D. Mechanisms of multidrug resistance in Acinetobacter species and Pseudomonas aeruginosa. Clin infect Dis 206; 43(2), S49-S56.
Clinical Laboratory and Standards Institute (CLSI). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard â€“ tenth edition. CLSI document M07-1. A10 Villanova, PA, USA: CLSI,. 2016.
Spilker T, Coenye T, Vandamme P, LiPuma JJ. PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. J Clini Microbial 2004;42 (5):2074-2079.
Pitout, J.D., Chow,B.L.,Gregson, D.B., Laupland, K.B., Elsayed, S. and Church, D.L.,2007. Molecular epidemiology of metallo-Î²-lactamase-producing Pseudomonas aeruginosa in the Calgary Health Region: emergence of VIM2-producing isolates. J Clin Microbiol 2007; 45(2), pp.294-298
Jiang X , Ni Y, Jiang Y, Yuan F, Han L, Li M, et al. Outbreak of infection caused by Enterobacter cloacae producing the novel VEB-3 beta-lactamase in China. J Clin Microbiol 2000; 43(2):826-31.
Fazeli N, Momtaz H. Virulence gene profiles of multidrugresistant Pseudomonas aeruginosa isolated from Iranian hospital infections. Iran Red Crescent Med J. 2014;18; 16 (12).
Shacheraghi F, Shakibaie MR, Noveiri H. Molecular Identification of ESBL Genes blaGES-1, blaVEB-1, blaCTX-M blaOXA-1, blaOXA-4, blaOXA-10 and blaPER-1in Pseudomonas aeruginosa Strains Isolated from Burn Patients by PCR, RFLP and Sequencing Techniques. W Acad Sci Eng Technol 2010; .4(1)2010.
Pitout JDD, Gregson DB, Poirel L, McClure J-A., Le P, Church DL. Detection of Pseudomonas aeruginosa Producing Metallo-Î²-Lactamases in a Large Centralized Laboratory. J Clin Microbiol 2005; 43(7): 3129â€“3135.
Akpaka PE, Swanston WH, Ihemere HN, Ihemere NH, Correa A, Torres A J. et al. Emergence of KPC-producing Pseudomonas aeruginosa in Trinidad and Tobago. J Clin Microbiol 2009;47(8):2670-1.
Wolska K, Szweda P. Genetic features of clinical Pseudomonas aeruginosa strains. Pol J Microbiol 2009;58:255-60.
Shaver CM, Hauser AR. Relative contributions of Pseudomonas aeruginosa ExoU, ExoS, and ExoT to virulence in the lung. Infect Immun 2004; 72(12), pp.6969-6977.
Mitov I, Strateva T, Markova B. Prevalence of virulence genes among Bulgarian nosocomial and cystic fibrosis isolates of Pseudomonas aeruginosa. Braz J Microbiol 2010; 41 (3):588-595.
Das T, Manefield M. Pyocyanin promotes extracellular DNA release in Pseudomonas aeruginosa. PLoS ONE , 2012; 7, e46718
Lau GW, Hassett DJ, Ran H, Kong F. The role of pyocyanin in Pseudomonas aeruginosa infection. Tren Mol Med 2004, 10, 599â€“606.
Hall S, McDermott C, Anoopkumar-Dukie, S, McFarland AJ, Forbes A. Perkins AV. Cellular Effects of Pyocyanin, a Secreted Virulence Factor of Pseudomonas aeruginosa. Toxins 2016, 8, 236.
Lister PD, Wolter DJ, Hanson ND. Antibacterial-Resistant Pseudomonas aeruginosa: Clinical Impact and Complex Regulation of Chromosomally Encoded Resistance Mechanisms. Clin Microbiol Rev 2009;22(4):582-610.
Abd Elmageed, MD AAE. Molecular Characterization and Sensitivity Patterns of Pseudomonas aeruginosa among patients with Different Diseases, PhD Thesis., Sudan University of Sciences and Technology, 2012.
Hussain M, Suliman M, Ahmed A, Altayb H, Elneima E. Draft Genome Sequence of a Multidrug-Resistant Pseudomonas aeruginosa Strain Isolated from a Patient with a Urinary Tract Infection in Khartoum, Sudan. Genome Announcements 2017;5(16):e00203-17. doi:10.1128/genomeA.00203-17.
Castanheira M, Deshpande LM, Costello A, Davies TA, Jones R. N. Epidemiology and carbapenem resistance mechanisms of carbapenem-non-susceptible Pseudomonas aeruginosa collected during 2009â€“11 in 14 European and Mediterranean countries. J Antimicrob Chemother 2014;69,1804â€“1814.
Poirel L, Nordmann P, Lagrutta E, Cleary T, and Munoz-Price LS. Emergence of KPC-producing Pseudomonas aeruginosa in the United States. Antimicrob Agents Chemother 2010; 54:3072.
Adler A, Navon-Venezia, S, Moran-Gilad J, Marcos E, Schwartz D, Carmeli Y. Laboratory and clinical evaluation of screening agar plates for detection of carbapenem-resistant Enterobacteriaceae from surveillance rectal swabs. J Clin Microbiol 2011; 49: 2239 42.
Agnello M, Wong-Beringer A. Differentiation in quinolone resistance by virulence genotype in Pseudomonas aeruginosa. PLoS One 2012; 7:e42973
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