Comparison of virulence factors between ESBL and non-ESBL producing Klebsiella pneumoniae isolates
Abstract
Background and Objectives: Klebsiella pneumoniae is an opportunistic pathogen responsible for causing nosocomial and community-acquired infections. Its pathogenicity is associated with a variety of virulence factors and antibiotic resistance. The aim of the present study was to compare virulence attributes between ESBL and non-ESBL producing isolates.
Materials and Methods: A total of 113 K. pneumoniae including 56 ESBL and 57 non ESBL-producers were collected in Bushehr province, Iran, from November 2017 to February 2019. Enzymatic profile, hypermucoviscosity and biofilm formation were investigated phenotypically. In addition, the presence of rmpA, aerobactin, kfu, allS, mrkD, ybtS, entB, iutA, fimH, wabG, wcaG, K1 and K2 genes were detected by PCR and sequencing.
Results: There was no statistically significant difference in enzymatic profile between ESBL and non-ESBL producers. The prevalence of the hypermocoviscosity was lower among ESBL compared to non-ESBL producers but the intensity of biofilm was higher in the ESBL producers. Among the virulence genes, K1, rmpA, iutA, and aero were observed only in non-ESBLs. Moreover, the carriage of allS, K, K2, rmpA, iutA and aero genes was higher in hypermucoviscous in comparison with non hypermucoviscous isolates.
Conclusion: The identification of potentially pathogenic isolates plays an important role in preventing their spread as well as the success of their treatment.
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2. Vuotto C, Longo F, Pascolini C, Donelli G, Balice MP, Libori MF, et al. Biofilm formation and antibiotic resistance in Klebsiella pneumoniae urinary strains. J Appl Microbiol 2017; 123: 1003-1018.
3. Cubero M, Marti S, Domínguez MÁ, González-Díaz A, Berbel D, Ardanuy C. Hypervirulent Klebsiella pneumoniae serotype K1 clinical isolates form robust biofilms at the air-liquid interface. PLoS One 2019; 14(9): e0222628.
4. Yu VL, Hansen DS, Ko WC, Sagnimeni A, Klugman KP, Von Gottberg A, et al. Virulence characteristics of Klebsiella and clinical manifestations of K. pneumoniae bloodstream infections. Emerg Infect Dis 2007; 13: 986-993.
5. Mirzaie A, Ranjbar R. Antibiotic resistance, virulence-associated genes analysis and molecular typing of Klebsiella pneumoniae strains recovered from clinical samples. AMB Express 2021; 11: 122.
6. Yu W-L, Fung C-P, Ko W-C, Chuang Y-C, Lee C-C, Chuang Y-C. Polymerase chain reaction analysis for detecting capsule serotypes K1 and K2 of Klebsiella pneumoniae causing abscesses of the liver and other sites. J Infect Dis 2007; 195: 1235-1236.
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8. Wasfi R, Elkhatib WF, Ashour HM. Molecular typing and virulence analysis of multidrug resistant Klebsiella pneumoniae clinical isolates recovered from Egyptian hospitals. Sci Rep 2016; 6: 38929.
9. Surgers L, Boyd A, Girard P-M, Arlet G, Decré D. Biofilm formation by ESBL-producing strains of Escherichia coli and Klebsiella pneumoniae. Int J Med Microbiol 2019; 309: 13-18.
10. Fu L, Huang M, Zhang X, Yang X, Liu Y, Zhang L, et al. Frequency of virulence factors in high biofilm formation blaKPC-2 producing Klebsiella pneumoniae strains from hospitals. Microb Pathog 2018; 116: 168-172.
11. Lee S, Han SW, Kim KW, Song DY, Kwon KT. Third-generation cephalosporin resistance of community-onset Escherichia coli and Klebsiella pneumoniae bacteremia in a secondary hospital. Korean J Intern Med 2014; 29: 49-56.
12. Thomas R, Hamat RA, Neela V. Extracellular enzyme profiling of Stenotrophomonas maltophilia clinical isolates. Virulence 2014; 5: 326-330.
13. Latifi B, Tajbakhsh S, Ahadi L, Yousefi F. Coexistence of aminoglycoside resistance genes in CTX-M-producing isolates of Klebsiella pneumoniae in Bushehr province, Iran. Iran J Microbiol 2021; 13: 161-170.
14. Barati A, Ghaderpour A, Chew LL, Bong CW, Thong KL, Chong VC, et al. Isolation and characterization of aquatic-borne Klebsiella pneumoniae from tropical estuaries in Malaysia. Int J Environ Res Public Health 2016; 13: 426.
15. Thong KL, Lai MY, Teh C SJ, Chua KH. Simultaneous detection of methicillin-resistant Staphylococcus aureus, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa by multiplex PCR. Trop Biomed 2011; 28: 21-31.
16. Wang T-C, Lin J-C, Chang J-C, Hiaso Y-W, Wang C-H, Chiu SK, et al. Virulence among different types of hypervirulent Klebsiella pneumoniae with multi-locus sequence type (MLST)-11, Serotype K1 or K2 strains. Gut Pathog 2021; 13: 40.
17. Kalaivani R, Jenitha JP, Seetha KS. Clinical isolates of Klebsiella pneumoniae and its virulence factors from a tertiary care hospital. Indian J Microbiol Res 2019; 6: 109-112.
18. Cevahir N, Demir M, Kaleli I, Gurbuz M, Tikvesli S. Evaluation of biofilm production, gelatinase activity, and mannose-resistant hemagglutination in Acinetobacter baumannii strains. J Microbiol Immunol Infect 2008; 41: 513-518.
19. Khalil MA, Ibrahim Sonbol F, Mohamed AF, Ali SS. Comparative study of virulence factors among ESβL-producing and nonproducing Pseudomonas aeruginosa clinical isolates. Turk J Med Sci 2015; 45: 60-69.
20. Stepanović S, Vuković D, Dakić I, Savić B, Švabić-Vlahović M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods 2000; 40: 175-179.
21. Donelli G, Vuotto C, Cardines R, Mastrantonio P. Biofilm-growing intestinal anaerobic bacteria. FEMS Immunol Med Microbiol 2012; 65: 318-325.
22. Lan Y, Zhou M, Jian Z, Yan Q, Wang S, Liu W. Prevalence of pks gene cluster and characteristics of Klebsiella pneumoniae‐induced bloodstream infections. J Clin Lab Anal 2019; 33(4): e22838.
23. Chiang T-T, Yang Y-S, Yeh K-M, Chiu S-K, Wang N-C, Lin T-Y, et al. Quantification and comparison of virulence and characteristics of different variants of carbapenemase-producing Klebsiella pneumoniae clinical isolates from Taiwan and the United States. J Microbiol Immunol Infect 2016; 49: 83-90.
24. Swathi CH, Chikala R, Ratnakar KS, Sritharan V. A structural, epidemiological & genetic overview of Klebsiella pneumoniae carbapenemases (KPCs). Indian J Med Res 2016; 144: 21-31.
25. Gundogan N, Citak S, Yalcin E. Virulence properties of extended spectrum β-lactamase-producing Klebsiella species in meat samples. J Food Prot 2011; 74: 559-564.
26. Pereira SC, Vanetti MC. Potential virulence of Klebsiella sp. isolates from enteral diets. Braz J Med Biol Res 2015; 48: 782-789.
27. Hennequin C, Robin F. Correlation between antimicrobial resistance and virulence in Klebsiella pneumoniae. Eur J Clin Microbiol Infect Dis 2016; 35: 333-341.
28. Yu W-L, Lee M-F, Tang H-J, Chang M-C, Chuang Y-C. Low prevalence of rmpA and high tendency of rmpA mutation correspond to low virulence of extended spectrum β-lactamase-producing Klebsiella pneumoniae isolates. Virulence 2015; 6: 162-172.
29. Rastegar S, Moradi M, Kalantar-Neyestanaki D, Ali Golabi D, Hosseini-Nave H. Virulence factors, capsular serotypes and antimicrobial resistance of hypervirulent Klebsiella pneumoniae and classical Klebsiella pneumoniae in Southeast Iran. Infect Chemother 2019; 10.3947/ic.2019.0027.
30.Taraghian A, Nasr Esfahani B, Moghim S, Fazeli H. Characterization of hypervirulent extended-spectrum β-lactamase-producing Klebsiella pneumoniae among urinary tract infections: the first report from Iran. Infect Drug Resist 2020; 13: 3103-3111.
31. Shadkam S, Goli HR, Mirzaei B, Gholami M, Ahanjan M. Correlation between antimicrobial resistance and biofilm formation capability among Klebsiella pneumoniae strains isolated from hospitalized patients in Iran. Ann Clin Microbiol Antimicrob 2021; 20: 13.
32. El Fertas-Aissani R, Messai Y, Alouache S, Bakour R. Virulence profiles and antibiotic susceptibility patterns of Klebsiella pneumoniae strains isolated from different clinical specimens. Pathol Biol (Paris) 2013; 61: 209-216.
33. Hasani A, Soltani E, Ahangarzadeh Rezaee M, Pirzadeh T, Ahangar Oskouee M, Hasani A, et al. Serotyping of Klebsiella pneumoniae and its relation with capsule-associated virulence genes, antimicrobial resistance pattern, and clinical infections: a descriptive study in medical practice. Infect Drug Resist 2020; 13: 1971-1980.
34. Bakhtiari R, Javadi A, Aminzadeh M, Molaee-Aghaee E, Shaffaghat Z. Association between presence of rmpA, mrkA and mrkD Genes and antibiotic resistance in clinical Klebsiella pneumoniae isolates from hospitals in Tehran, Iran. Iran J Public Health 2021; 50: 1009-1016.
35. Dan B, Dai H, Zhou D, Tong H, Zhu M. Relationship between drug resistance characteristics and biofilm formation in Klebsiella Pneumoniae strains. Infect Drug Resist 2023; 16: 985-998.
36. Jung SW, Chae HJ, Park YJ, Yu JK, Kim SY, Lee HK, et al. Microbiological and clinical characteristics of bacteraemia caused by the hypermucoviscosity phenotype of Klebsiella pneumoniae in Korea. Epidemiol Infect 2013; 141: 334-340.
37. Yu W-L, Ko W-C, Cheng K-C, Lee H-C, Ke D-S, Lee C-C, et al. Association between rmpA and magA genes and clinical syndromes caused by Klebsiella pneumoniae in Taiwan. Clin Infect Dis 2006; 42: 1351-1358.
38. Fu Y, Xu M, Liu Y, Li A, Zhou J. Virulence and genomic features of a blaCTX-M-3 and blaCTX-M-14 coharboring hypermucoviscous Klebsiella pneumoniae of serotype K2 and ST65. Infect Drug Resist 2019; 12: 145-159.
39. Soltani E, Hasani A, Rezaee MA, Pirzadeh T, Oskouee MA, Hasani A, et al. Virulence characterization of Klebsiella pneumoniae and its relation with ESBL and AmpC beta-lactamase associated resistance. Iran J Microbiol 2020; 12: 98-106.
40. Bachman MA, Oyler JE, Burns SH, Caza M, Lépine F, Dozois CM, et al. Klebsiella pneumoniae yersiniabactin promotes respiratory tract infection through evasion of lipocalin 2. Infect Immun 2011; 79: 3309-3316.
2. Vuotto C, Longo F, Pascolini C, Donelli G, Balice MP, Libori MF, et al. Biofilm formation and antibiotic resistance in Klebsiella pneumoniae urinary strains. J Appl Microbiol 2017; 123: 1003-1018.
3. Cubero M, Marti S, Domínguez MÁ, González-Díaz A, Berbel D, Ardanuy C. Hypervirulent Klebsiella pneumoniae serotype K1 clinical isolates form robust biofilms at the air-liquid interface. PLoS One 2019; 14(9): e0222628.
4. Yu VL, Hansen DS, Ko WC, Sagnimeni A, Klugman KP, Von Gottberg A, et al. Virulence characteristics of Klebsiella and clinical manifestations of K. pneumoniae bloodstream infections. Emerg Infect Dis 2007; 13: 986-993.
5. Mirzaie A, Ranjbar R. Antibiotic resistance, virulence-associated genes analysis and molecular typing of Klebsiella pneumoniae strains recovered from clinical samples. AMB Express 2021; 11: 122.
6. Yu W-L, Fung C-P, Ko W-C, Chuang Y-C, Lee C-C, Chuang Y-C. Polymerase chain reaction analysis for detecting capsule serotypes K1 and K2 of Klebsiella pneumoniae causing abscesses of the liver and other sites. J Infect Dis 2007; 195: 1235-1236.
7. Yu W-L, Ko W-C, Cheng K-C, Lee C-C, Lai C-C, Chuang Y-C. Comparison of prevalence of virulence factors for Klebsiella pneumoniae liver abscesses between isolates with capsular K1/K2 and non-K1/K2 serotypes. Diagn Microbiol Infect Dis 2008; 62: 1-6.
8. Wasfi R, Elkhatib WF, Ashour HM. Molecular typing and virulence analysis of multidrug resistant Klebsiella pneumoniae clinical isolates recovered from Egyptian hospitals. Sci Rep 2016; 6: 38929.
9. Surgers L, Boyd A, Girard P-M, Arlet G, Decré D. Biofilm formation by ESBL-producing strains of Escherichia coli and Klebsiella pneumoniae. Int J Med Microbiol 2019; 309: 13-18.
10. Fu L, Huang M, Zhang X, Yang X, Liu Y, Zhang L, et al. Frequency of virulence factors in high biofilm formation blaKPC-2 producing Klebsiella pneumoniae strains from hospitals. Microb Pathog 2018; 116: 168-172.
11. Lee S, Han SW, Kim KW, Song DY, Kwon KT. Third-generation cephalosporin resistance of community-onset Escherichia coli and Klebsiella pneumoniae bacteremia in a secondary hospital. Korean J Intern Med 2014; 29: 49-56.
12. Thomas R, Hamat RA, Neela V. Extracellular enzyme profiling of Stenotrophomonas maltophilia clinical isolates. Virulence 2014; 5: 326-330.
13. Latifi B, Tajbakhsh S, Ahadi L, Yousefi F. Coexistence of aminoglycoside resistance genes in CTX-M-producing isolates of Klebsiella pneumoniae in Bushehr province, Iran. Iran J Microbiol 2021; 13: 161-170.
14. Barati A, Ghaderpour A, Chew LL, Bong CW, Thong KL, Chong VC, et al. Isolation and characterization of aquatic-borne Klebsiella pneumoniae from tropical estuaries in Malaysia. Int J Environ Res Public Health 2016; 13: 426.
15. Thong KL, Lai MY, Teh C SJ, Chua KH. Simultaneous detection of methicillin-resistant Staphylococcus aureus, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa by multiplex PCR. Trop Biomed 2011; 28: 21-31.
16. Wang T-C, Lin J-C, Chang J-C, Hiaso Y-W, Wang C-H, Chiu SK, et al. Virulence among different types of hypervirulent Klebsiella pneumoniae with multi-locus sequence type (MLST)-11, Serotype K1 or K2 strains. Gut Pathog 2021; 13: 40.
17. Kalaivani R, Jenitha JP, Seetha KS. Clinical isolates of Klebsiella pneumoniae and its virulence factors from a tertiary care hospital. Indian J Microbiol Res 2019; 6: 109-112.
18. Cevahir N, Demir M, Kaleli I, Gurbuz M, Tikvesli S. Evaluation of biofilm production, gelatinase activity, and mannose-resistant hemagglutination in Acinetobacter baumannii strains. J Microbiol Immunol Infect 2008; 41: 513-518.
19. Khalil MA, Ibrahim Sonbol F, Mohamed AF, Ali SS. Comparative study of virulence factors among ESβL-producing and nonproducing Pseudomonas aeruginosa clinical isolates. Turk J Med Sci 2015; 45: 60-69.
20. Stepanović S, Vuković D, Dakić I, Savić B, Švabić-Vlahović M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods 2000; 40: 175-179.
21. Donelli G, Vuotto C, Cardines R, Mastrantonio P. Biofilm-growing intestinal anaerobic bacteria. FEMS Immunol Med Microbiol 2012; 65: 318-325.
22. Lan Y, Zhou M, Jian Z, Yan Q, Wang S, Liu W. Prevalence of pks gene cluster and characteristics of Klebsiella pneumoniae‐induced bloodstream infections. J Clin Lab Anal 2019; 33(4): e22838.
23. Chiang T-T, Yang Y-S, Yeh K-M, Chiu S-K, Wang N-C, Lin T-Y, et al. Quantification and comparison of virulence and characteristics of different variants of carbapenemase-producing Klebsiella pneumoniae clinical isolates from Taiwan and the United States. J Microbiol Immunol Infect 2016; 49: 83-90.
24. Swathi CH, Chikala R, Ratnakar KS, Sritharan V. A structural, epidemiological & genetic overview of Klebsiella pneumoniae carbapenemases (KPCs). Indian J Med Res 2016; 144: 21-31.
25. Gundogan N, Citak S, Yalcin E. Virulence properties of extended spectrum β-lactamase-producing Klebsiella species in meat samples. J Food Prot 2011; 74: 559-564.
26. Pereira SC, Vanetti MC. Potential virulence of Klebsiella sp. isolates from enteral diets. Braz J Med Biol Res 2015; 48: 782-789.
27. Hennequin C, Robin F. Correlation between antimicrobial resistance and virulence in Klebsiella pneumoniae. Eur J Clin Microbiol Infect Dis 2016; 35: 333-341.
28. Yu W-L, Lee M-F, Tang H-J, Chang M-C, Chuang Y-C. Low prevalence of rmpA and high tendency of rmpA mutation correspond to low virulence of extended spectrum β-lactamase-producing Klebsiella pneumoniae isolates. Virulence 2015; 6: 162-172.
29. Rastegar S, Moradi M, Kalantar-Neyestanaki D, Ali Golabi D, Hosseini-Nave H. Virulence factors, capsular serotypes and antimicrobial resistance of hypervirulent Klebsiella pneumoniae and classical Klebsiella pneumoniae in Southeast Iran. Infect Chemother 2019; 10.3947/ic.2019.0027.
30.Taraghian A, Nasr Esfahani B, Moghim S, Fazeli H. Characterization of hypervirulent extended-spectrum β-lactamase-producing Klebsiella pneumoniae among urinary tract infections: the first report from Iran. Infect Drug Resist 2020; 13: 3103-3111.
31. Shadkam S, Goli HR, Mirzaei B, Gholami M, Ahanjan M. Correlation between antimicrobial resistance and biofilm formation capability among Klebsiella pneumoniae strains isolated from hospitalized patients in Iran. Ann Clin Microbiol Antimicrob 2021; 20: 13.
32. El Fertas-Aissani R, Messai Y, Alouache S, Bakour R. Virulence profiles and antibiotic susceptibility patterns of Klebsiella pneumoniae strains isolated from different clinical specimens. Pathol Biol (Paris) 2013; 61: 209-216.
33. Hasani A, Soltani E, Ahangarzadeh Rezaee M, Pirzadeh T, Ahangar Oskouee M, Hasani A, et al. Serotyping of Klebsiella pneumoniae and its relation with capsule-associated virulence genes, antimicrobial resistance pattern, and clinical infections: a descriptive study in medical practice. Infect Drug Resist 2020; 13: 1971-1980.
34. Bakhtiari R, Javadi A, Aminzadeh M, Molaee-Aghaee E, Shaffaghat Z. Association between presence of rmpA, mrkA and mrkD Genes and antibiotic resistance in clinical Klebsiella pneumoniae isolates from hospitals in Tehran, Iran. Iran J Public Health 2021; 50: 1009-1016.
35. Dan B, Dai H, Zhou D, Tong H, Zhu M. Relationship between drug resistance characteristics and biofilm formation in Klebsiella Pneumoniae strains. Infect Drug Resist 2023; 16: 985-998.
36. Jung SW, Chae HJ, Park YJ, Yu JK, Kim SY, Lee HK, et al. Microbiological and clinical characteristics of bacteraemia caused by the hypermucoviscosity phenotype of Klebsiella pneumoniae in Korea. Epidemiol Infect 2013; 141: 334-340.
37. Yu W-L, Ko W-C, Cheng K-C, Lee H-C, Ke D-S, Lee C-C, et al. Association between rmpA and magA genes and clinical syndromes caused by Klebsiella pneumoniae in Taiwan. Clin Infect Dis 2006; 42: 1351-1358.
38. Fu Y, Xu M, Liu Y, Li A, Zhou J. Virulence and genomic features of a blaCTX-M-3 and blaCTX-M-14 coharboring hypermucoviscous Klebsiella pneumoniae of serotype K2 and ST65. Infect Drug Resist 2019; 12: 145-159.
39. Soltani E, Hasani A, Rezaee MA, Pirzadeh T, Oskouee MA, Hasani A, et al. Virulence characterization of Klebsiella pneumoniae and its relation with ESBL and AmpC beta-lactamase associated resistance. Iran J Microbiol 2020; 12: 98-106.
40. Bachman MA, Oyler JE, Burns SH, Caza M, Lépine F, Dozois CM, et al. Klebsiella pneumoniae yersiniabactin promotes respiratory tract infection through evasion of lipocalin 2. Infect Immun 2011; 79: 3309-3316.
| Files | ||
| Issue | Vol 16 No 1 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v16i1.14869 | |
| Keywords | ||
| Klebsiella pneumoniae; Biofilm; Extended-spectrum beta-lactamase; Virulence | ||
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How to Cite
1.
Hatami Mirbag H, Tajbakhsh S, Askari A, Yousefi F. Comparison of virulence factors between ESBL and non-ESBL producing Klebsiella pneumoniae isolates. Iran J Microbiol. 2024;16(1):39-48.
