Prevalence and leukotoxic profile of biofilm forming Staphylococcal strains isolated from a tertiary care hospital in Mysore
Keywords:
Biofilm, Device-associated infections, Staphylococcus, Pantone-Valentine Leukocidin, ica operon
Abstract
Background: Staphylococci causes chronic skin infections, device associated infections, ear infections, bone infections and post-operative infections. It secretes toxins such as Pantone-Valentine Leukocidin which prolongs the infection in hospitalized patients.
Aim: To investigate the prevalence of Staphylococcal biofilm associated strains and to understand the correlation between biofilm production, multi-drug resistance and leukotoxicity.
Methods: 280 Staphylococcal clinical isolates obtained from biofilm associated infections are collected from a tertiary care hospital in Mysore. They were screened for the production of biofilm, multi-drug resistance and leukotoxicity. They were also screened for icaAB and PVL luk F/S genes by PCR. Correlation between leuktoxicity and biofilm production was established by finding the leukotoxic potential of biofilm positive strains.
Findings: Among 280 Staphylococcal clinical isolates, 34.6% showed biofilm production. Out of these isolates, 25% showed the presence of icaAB genes and 29% showed the presence of PVL luk F/S genes. Presence of icaAB gene was found to be higher in isolates obtained from implant associated infections. But the prevalence of PVL luk F/S genes was higher in chronic skin infections. 78% of the isolates displayed multi-drug resistance and methicillin resistant strains were comparitively high in device associated infections (77%). Strong biofilm producing PVL positive isolates from chronic skin infections showed increased leucotoxicity compared to PVL negative, weak and moderate biofilm producers.
Conclusion: Statistical analysis revealed that PVL in association with biofilms can promote higher leukotoxicity compared to non biofilm producers. This study foresee the essential role of biofilm production, multi-drug resistance and toxins for the increased Staphylococcal pathogenesis.
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References
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31. Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods. 2000;40(2):175-9.
32. Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966 Apr;45(4):493-6.
33. Clinical Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; Twenty Second Informational Supplement. 2012;32(3). Document M100-S22, Wayne, PA.
34. Frebourg NB, Lefebvre S, Baert S, Lemeland JF. PCR-Based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains. J Clin Microbiol. 2000 Feb;38(2):877-80.
35. Lina G, Piémont Y, Godail-Gamot F, Bes M, Peter MO, Gauduchon V, Vandenesch F, Etienne J. Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis. 1999 Nov;29(5):1128-32.
36. Chung WB, Bäckström LR, McDonald J, Collins MT. The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) colorimetric assay for the quantitation of Actinobacillus pleuropneumoniae cytotoxin. Can J Vet Res. 1993 Jul;57(3):159-65. [PubMed]
37. Padmaja RJ, Halami PM. Molecular Characterization and Toxicity Confirmation of LukM/F'-PV Producing Staphylococcus aureus Isolated from Bovine Mastitis Samples in Mysore, India. Indian Journal of Microbiology. 2013;53(3):276-82.
38. Deepa S, Kumari A, Venkatesha D. Increasing trends of methicillin resistant coagulase negative Staphylococcus in neonatal septicaemia – a study in a tertiary care hospital, Mysore, South India. Online Journal of Health and Allied Sciences 2010, 9(4):1-3.
39. Annu T, Sumana M, Umamaheshwari S. Screening health care personnel for detection of methicillin resistant of Staphylococcus aureus carrier state at a tertiary care hospital in Mysore. International Journal of Pharmaceutical Science Invention 2013;2(7):12-15.
40. Lyra PR, Anuradha K, Shilpa A, Venkatesha D. Linezolid resistance in isolates of methicillin resistant Staphylococci from blood cultures. International Journal of Pharma and Biosciences 2013; 4(4):1085-1090.
2. von Eiff C, Jansen B, Kohnen W, Becker K. Infections associated with medical devices: pathogenesis, management and prophylaxis. Drugs. 2005;65(2):179-214. [PubMed]
3. McCann MT, Gilmore BF, Gorman SP. Staphylococcus epidermidis device-related infections: pathogenesis and clinical management. Journal of Pharmacy and Pharmacology. 2008;60(12):1551-71.
4. Costerton JW, Montanaro L, Arciola CR. Biofilm in implant infections: its production and regulation. Int J Artif Organs. 2005 Nov;28(11):1062-8. [PubMed]
5. Donlan RM. Biofilms and device-associated infections. Emerg Infect Dis. 2001 Mar-Apr;7(2):277-81. [PubMed]
6. Hooper S, Percival S, Cochrane C, Williams D. Biofilms and Implication in Medical Devices in Humans and Animals. In: Percival S, Knottenbelt D, Cochrane C, editors. Biofilms and Veterinary Medicine. Springer Series on Biofilms. 6: Springer Berlin Heidelberg; 2011. p. 191-203.
7. Murugan K, Usha M, Malathi P, Al-Sohaibani AS, Chandrasekaran M. Biofilm forming multi drug resistant Staphylococcus spp. among patients with conjunctivitis. Pol J Microbiol. 2010;59(4):233-9. [PubMed]
8. O'Grady NP, Alexander M, Burns LA, Dellinger EP, Garland J, Heard SO, et al. Guidelines for the prevention of intravascular catheter-related infections. American Journal of Infection Control. 2011;39(4):S1-S34.
9. Gilbert P, Das J, Foley I. Biofilm susceptibility to antimicrobials. Adv Dent Res. 1997 Apr;11(1):160-7. [PubMed]
10. Ceri H, Olson ME, Stremick C, Read RR, Morck D, Buret A. The Calgary Biofilm Device: new technology for rapid determination of antibiotic susceptibilities of bacterial biofilms. J Clin Microbiol. 1999 Jun;37(6):1771-6.
11. Heilmann C, Schweitzer O, Gerke C, Vanittanakom N, Mack D, Götz F. Molecular basis of intercellular adhesion in the biofilm-forming Staphylococcus epidermidis. Molecular Microbiology. 1996;20(5):1083-91.
12. Ziebuhr W, Heilmann C, Götz F, Meyer P, Wilms K, Straube E, Hacker J. Detection of the intercellular adhesion gene cluster (ica) and phase variation in Staphylococcus epidermidis blood culture strains and mucosal isolates. Infect Immun. 1997 Mar;65(3):890-6.
13. Kathju S, Nistico L, Hall-Stoodley L, Post JC, Ehrlich GD, Stoodley P. Chronic Surgical Site Infection Due to Suture-Associated Polymicrobial Biofilm. Surgical Infections. 2009;10(5):457-61.
14. Pulcrano G, Vollaro A, Rossano F, Catania MR. Molecular and Phenotypic Characterization of Methicillin-Resistant Staphylococcus aureus from Surgical Site Infections. Surgical Infections. 2013;14(2):196-202.
15. Metcalf DG, Bowler PG. Biofilm delays wound healing: A review of the evidence. Burns Trauma. 2013 Jun 18;1(1):5-12. doi: 10.4103/2321-3868.113329. eCollection 2013. [PubMed]
16. Cheng AG, DeDent AC, Schneewind O, Missiakas D. A play in four acts: Staphylococcus aureus abscess formation. Trends Microbiol. 2011 May;19(5):225-32. doi: 10.1016/j.tim.2011.01.007. Epub 2011 Feb 25. [PubMed]
17. Brady RA, Leid JG, Calhoun JH, Costerton JW, Shirtliff ME. Osteomyelitis and the role of biofilms in chronic infection. FEMS Immunol Med Microbiol. 2008 Jan;52(1):13-22. Epub 2007 Dec 11. [PubMed]
18. Aktas NC, Erturan Z, Karatuna O, Yagci AK. Panton-Valentine leukocidin and biofilm production of Staphylococcus aureus isolated from respiratory tract. Journal of Infection in Developing Countries. 2013;7(11):888-91.
19. Paluch-Oleś J, Magryś A, Kozioł-Montewka M, Niedzielski A, Niedźwiadek J, Niedzielska G, Kotowski M. The phenotypic and genetic biofilm formation characteristics of coagulase-negative staphylococci isolates in children with otitis media. Int J Pediatr Otorhinolaryngol. 2011 Jan;75(1):126-30. doi: 10.1016/j.ijporl.2010.10.025. Epub 2010 Nov 18.
20. Ando E, Monden K, Mitsuhata R, Kariyama R, Kumon H. Biofilm formation among methicillin-resistant Staphylococcus aureus isolates from patients with urinary tract infection. Acta Medica Okayama. 2004;58(4):207-14.
21. Arciola CR, Alvi FI, An YH, Campoccia D, Montanaro L. Implant infection and infection resistant materials: A mini review. International Journal of Artificial Organs. 2005;28(11):1119-25.
22. Singhai M, Malik A, Shahid M, Malik MA, Goyal R. A study on device-related infections with special reference to biofilm production and antibiotic resistance. J Glob Infect Dis. 2012;4(4):193-8.
23. Cosgrove SE, Qi YL, Kaye KS, Harbarth S, Karchmer AW, Carmeli Y. The impact of methicillin-resistance in Staphylococcus aureus bacteremia on patient outcomes: Mortality, length of stay, and hospital charges. Infection Control and Hospital Epidemiology. 2005;26(2):166-74.
24. Agarwal A, Jain A. Association between drug resistance & production of biofilm in staphylococci. Indian Journal of Medical Research. 2012;135(4):562-4. [PubMed]
25. Kwon AS, Park GC, Ryu SY, Lim DH, Lim DY, Choi CH, Park Y, Lim Y. Higher biofilm formation in multidrug-resistant clinical isolates of Staphylococcus aureus. Int J Antimicrob Agents. 2008 Jul;32(1):68-72. doi: 10.1016/j.ijantimicag.2008.02.009. Epub 2008 Jun 5.
26. Pozzi C, Waters EM, Rudkin JK, Schaeffer CR, Lohan AJ, Tong P, et al. Methicillin Resistance Alters the Biofilm Phenotype and Attenuates Virulence in Staphylococcus aureus Device-Associated Infections. Plos Pathogens. 2012;8(4).
27. Etienne J. Panton-Valentine leukocidin: a marker of severity for Staphylococcus aureus infection? Clin Infect Dis. 2005 Sep 1;41(5):591-3. Epub 2005 Jul 29. [PubMed]
28. Szmigielski S, Prevost G, Monteil H, Colin DA, Jeljaszewicz J. Leukocidal toxins of staphylococci. Zentralblatt Fur Bakteriologie-International Journal of Medical Microbiology Virology Parasitology and Infectious Diseases. 1999;289(2):185-201.
29. Prevost G, Cribier B, Couppie P, Petiau P, Supersac G, Finckbarbancon V, et al. Panton-Valentine Leucocidin and gamma-hemolysin from Staphylococcus aureus ATCC-49775 are encoded by distinct genetic-loci and have different biological activities. Infection and Immunity. 1995;63(10):4121-9.
30. Lo WT, Wang CC. Panton-Valentine Leukocidin in the Pathogenesis of Community-associated Methicillin-resistant Staphylococcus aureus Infection. Pediatrics and Neonatology. 2011;52(2):59-65.
31. Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods. 2000;40(2):175-9.
32. Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966 Apr;45(4):493-6.
33. Clinical Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; Twenty Second Informational Supplement. 2012;32(3). Document M100-S22, Wayne, PA.
34. Frebourg NB, Lefebvre S, Baert S, Lemeland JF. PCR-Based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains. J Clin Microbiol. 2000 Feb;38(2):877-80.
35. Lina G, Piémont Y, Godail-Gamot F, Bes M, Peter MO, Gauduchon V, Vandenesch F, Etienne J. Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis. 1999 Nov;29(5):1128-32.
36. Chung WB, Bäckström LR, McDonald J, Collins MT. The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) colorimetric assay for the quantitation of Actinobacillus pleuropneumoniae cytotoxin. Can J Vet Res. 1993 Jul;57(3):159-65. [PubMed]
37. Padmaja RJ, Halami PM. Molecular Characterization and Toxicity Confirmation of LukM/F'-PV Producing Staphylococcus aureus Isolated from Bovine Mastitis Samples in Mysore, India. Indian Journal of Microbiology. 2013;53(3):276-82.
38. Deepa S, Kumari A, Venkatesha D. Increasing trends of methicillin resistant coagulase negative Staphylococcus in neonatal septicaemia – a study in a tertiary care hospital, Mysore, South India. Online Journal of Health and Allied Sciences 2010, 9(4):1-3.
39. Annu T, Sumana M, Umamaheshwari S. Screening health care personnel for detection of methicillin resistant of Staphylococcus aureus carrier state at a tertiary care hospital in Mysore. International Journal of Pharmaceutical Science Invention 2013;2(7):12-15.
40. Lyra PR, Anuradha K, Shilpa A, Venkatesha D. Linezolid resistance in isolates of methicillin resistant Staphylococci from blood cultures. International Journal of Pharma and Biosciences 2013; 4(4):1085-1090.
CITATION
DOI: 10.17511/jopm.2016.i03.06
Published: 2016-12-31
How to Cite
Halebeedu Prakash, P., & Gopal, S. (2016). Prevalence and leukotoxic profile of biofilm forming Staphylococcal strains isolated from a tertiary care hospital in Mysore. Tropical Journal of Pathology and Microbiology, 2(3), 110-119. https://doi.org/10.17511/jopm.2016.i03.06
Section
Original Article
OAI - Open Archives Initiative
