Identification and Antibiotic Resistance Profile of Biofilm-forming Methicillin Resistant Staphylococcus aureus (MRSA) Causing Infection among Orthopedic Wound Patients
Asian Journal of Research in Medical and Pharmaceutical Sciences,
Background and Objectives: The biofilm-forming ability of Methicillin-Resistant Staphylococcus aureus(MRSA) strains have demonstrated the involvement of MRSA biofilm in antibiotic resistance, recalcitrant and persistent infections in humans. Despite a deeper understanding of the biofilm-forming ability of MRSAstrain, it is still essential to extend the research on the identification and antibiotic resistance profile of biofilm-forming MRSA causing infection among orthopedic wound patients.
Methodology: A total of three hundred and thirty (303) patient-isolate of non-repeatable Staphylococcus aureus strains were obtained during the period of 2021 until 2022 from fracture and post-surgical orthopedic wound patients with wound duration >2months at the National Orthopedic Hospital, Enugu (NOHE). S. aureus were identified using conventional microbiological cultures Technique followed by confirmation of MRSA strain through Brilliance MRSA 2 Agar. Antibiotic Susceptibility testing (AST) of biofilm-forming MRSA was performed using the Kirby–Bauer disk diffusion method and the results were interpreted using the Clinical Laboratory Standard Institute (CLSI) zone diameter breakpoints. Multidrug Resistance (MDR) was determined for biofilm-forming MRSA.
Result:Of the 303 isolate of S. aureus, MRSA strain accounted 86(28.4 %) and 78(25.7 %) from post-surgical wound and fracture wound respectively while biofilm forming MRSA was identified in 101(33.4%) MRSA strain consisting of high proportion 66(21.8 %) fromPost-surgical wound followed by fracture wound samples recording 35(11.6 %). Association between MRSA production and biofilm formation was considered statistically significant at P< .05. The proportion of biofilm-forming MRSA resistance to β-lactam accounted 71.4-100%, macrolide resistance recorded 65.7-92.4 %, lincosamideresistance 74.3-100 %, glycopeptide resistance proportion ranged from 62.8-100 % while low level of resistance to fluoroquinolones 19.7-42.9 % and Aminoglycoside 8.6-10.6 % was observed. Biofilm-forming MRSA isolate were MDR to one or more antibiotic antimicrobial agents in at least three categories withMDRIndex range ≥ 0.3 but majority of the isolate were 91.4% and 100% susceptible to Gentamicin and Imipenem.
Conclusion: The invitro expression of biofilm formation among MRSA strain and their antibiotic resistance profile in this study makes them a potential threat and challenging pathogens with the ability to cause persistent infections in humans, especially among orthopedic wound patients. Thus the development of an antimicrobial stewardship program and regular detection of biofilm production is needed for timely intervention while judicious use of Imipenem and Gentamicin as a drug of choice for effective treatment of infection caused by biofilm-forming MRSA among orthopedic patients will avert the severity of infection. Further research of these sort should investigate the genotyping expression of a biofilm gene variant in other human diseases, different bacteria species, and orthopedic medical implant devices.
- Methicillin Resistant Staphylococcus aureus
- antibiotic resistance
How to Cite
Silva V, Almeida L, Gaio V, Cerca N, Manageiro V, Caniça M, Capelo JL, Igrejas G, Poeta P. Biofilm formation of multidrug resistant MRSA strains isolated from different types of human infections. Pathogens. 2021;10:9-70.
Peter IU, Ngwu JN, Edemekong CI, Ugwueke IV, Uzoeto HO, Joseph OV, Mohammed ID, Mbong EO, Nomeh OL, Ikusika BA, Ubom IJ, Inyogu JC, Ntekpe ME, Obodoechi IF, NseAbasi, PL, Ogbonna IP, Didiugwu CM, Akpu PO, Alagba EE, Ogba RC, Iroha IR. First report prevalence of livestock acquired Methicillin Resistant Staphylococcus aureus (LA-MRSA) strain in South Eastern, Nigeria . IOSR J Nurs Health Sci. 2022;11(1):50-56.
Silva V, Almeida F, Carvalho JA, Castro AP, Ferreira E, Manageiro V, Tejedor-Junco MT, Caniça M, Igrejas G, Poeta P. Emergence of community-acquired Methicillin Resistant Staphylococcus aureus EMRSA-15 clone as the predominant cause of diabetic foot ulcer infections in Portugal.European J ClinMicrobiol Infect Dis. 2020;39:179–186.
Haddad O, Merghni A, Elargoubi A, Rhim H, Kadri Y, Mastouri M. Comparative study of virulence factors among Methicillin Resistant Staphylococcus aureus clinical isolates. BMC Infect Dis.2018;18:560-456.
Haysom L, Cross M, Anastasas R, Moore E, Hampton S. Prevalence and risk factors for Methicillin-Resistant Staphylococcus aureus (MRSA) infections in custodial populations: A systematic review. J Corr Health Care. 2018;24:197–213.
Abdulrahim U, Kachallah M, Rabiu M, Usman NA, Adeshina GO, Olayinka BO. Molecular detection of biofilm-producing Staphylococcus aureus isolates from national orthopaedic Hospital Dala, Kano State, Nigeria. Open J Med Microbiol. 2019;9:116-126.
Puca V, Marulli RZ, Grande R, Vitale I, Niro A, Molinaro G, Prezioso S, Muraro R, Di Giovanni P. Microbial species isolated from infected wounds and antimicrobial resistance analysis: data emerging from a three-years retrospective study. Antibiotics. 2021;10:1162.
Garoy EY, Gebreab YB, Achila OO, Tekeste DG, Robel RK, Kiflay GR, Tesfu T. Methicillin-Resistant Staphylococcus aureus(MRSA): Prevalence and antimicrobial sensitivity pattern among patients-a multicenter study in Asmara, Eritrea. Can J Infect Dis Med Microbiol.2018;9:832-1834.
Coello R, Jimenez J, Garcia M. Prospective study of infection, colonization and carriage of methicillin-resistant Staphylococcus aureusin an outbreak affecting 990 patients. European Journal of ClinMicrobiol Infect Dis. 1994;13(1):74–81.
Coraça-Huber DC, Kreid L, Steixner S, Hinz M, Dammerer D, Fille M. Identification and morphological characterization ofbiofilms formed by strains causing infection inorthopedic implants. Pathogens. 2020;9:6-49.
Cascioferro S, Carbone D, Parrino B, Pecoraro C, Giovannetti E, Cirrincione G, Diana P. Therapeutic strategies to counteract antibiotic resistance in MRSA biofilm-associated infections. Chemical and Med Chem. 2021;16:65–80.
Saeed K, McLaren AC, Schwarz EM, Antoci V, Arnold WV, Chen AF, Clauss M, Esteban J, Gant V, Hendershot E. 2018 International consensus meeting on musculoskeletal infection: Summary from the biofilm workgroup and consensus on biofilm related musculoskeletal infections. J. Orthop. Res. 2019;37:1007–1017.
Chang JH, Chong KKL, Lam LN, Wong JJ, Kline KA. Biofilm-associated Infection by Enterococci. Nature Rev Microbiol.2018;17:82–94.
Pinto RM, Soares FA, Reis S, Nunes C, Van Dijck P. Innovative strategies toward the disassembly of the EPS matrix in bacterial biofilms. Front Microbiol. 2020;11:952-967.
Stewart PS. Antimicrobial tolerance in biofilms. In microbiology spectrum; Ghannoum M, Parsek M, Whiteley M, Mukherjee PK, Eds.; American Society for Microbiology: Washington, DC, USA. 2015;3.
Hall CW, Mah TF. Molecular mechanisms of biofilm-based antibiotic resistance and tolerance in pathogenic bacteria. FEMS Microbiol. Rev. 2017;41(23):276–301.
Iroha IR, Orji JO, Onwa NC, Nwuzo AC, Okonkwo EC, Ibiam EO, Nwachi AC, Afuikwa FN, Agah VM, Ejikeugwu EPC, Agumah NB, Moses IB, Ugbo E, Ukpai EG, Nwakaeze E A, Oke B, Ogbu L, Nwunna E. Microbiology practical handbook. (Editor; Ogbu. O), 1st Edition. Charlieteximage Africa (CiAfrica Press), 2019;344.
Ngwu JN, Uzoeto HO, Emaimo J, Okorie C, Mohammed ID, Edemekong CI,Peter IU, Ezeh C, Chukwu E, Adimora EE, Ani SE, Oke B, Moses IB, Nwakaeze EA, Otu JO, Chukwunwejim CR, Egbuna RN, Ikusika BA, Adagiri P, Iroha IR. Antibiogram of biofilm forming oral Streptococci species isolated from dental caries patients visiting federal College of Dental Technology and Therapy, Enugu Nigeria. Int J Res Rep Dent. 2022;5(1):12-25.
Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing; twenty-eighth edition (M100). Wayne, PA: Clinical and Laboratory Standards Institute; 2019.
Fahim NAE. Prevalence and antimicrobial susceptibility profile of multidrug-resistant bacteria among intensive care units patients at Ain Shams University Hospitals in Egypt—A Retrospective Study. J Egypt Pub Health Assoc. 2021;96:7-34.
Ghaznavi-Rad E, Ekrami A. Molecular characterization of Methicillin-Resistant Staphylococcus aureusisolates, isolated from a Burn Hospital, in Southwest Iran in 2006 and 2014. HindawiInt J Microbiol. 2018;5(23):123-200.
Rebiahi SA, Abdelouahid DE, Rahmoun M, Abdelali S, Azzaoui H. Emergence of vancomycin-resistant Staphylococcus aureusidentified in the Tlemcen University Hospital (North-West Algeria). M´edecineet MaladiesInfectieuses. 2011;41(12):646–651.
Guzman-Blanco M, Carlos M, Raul I. Review epidemiology of Methicillin-resistant Staphylococcus aureus (MRSA) in Latin America. Int J Antimicrob Agents. 2009;34(4):304–308.
Jimenez JN, Ocampo AM, Vanegas JM. CC8 MRSA strains harboring SCCmec type IV are predominant in Colombian Hospitals. PLOS One. 2012;7(6):38-576.
Tsige Y, Senait TS, Eyesus GT, Tefera MM, Amsalu A, Menberu AM, Gelaw B. Prevalence of Methicillin-resistant Staphylococcus aureus and associated risk factors among patients with wound infection at Referral Hospital, Northeast Ethiopia. J Pathogens.2020;31(7):56-78.
Mawalla B, Mshana SE, Chalya PL, Imirzalioglu C, Mahalu W. Predictors of surgical site infections among patients undergoing major surgery at Bugando Medical Centre in Northwestern Tanzania. BMC Surg Infect.2011;11:21-23.
Anguzu JR, Olila D. Drug sensitivity patterns of bacterial isolates from septic post-operative wounds in a regional referral hospital in Uganda. Afr J Health Sci. 2007;7:148–154.
Ghanem S, Bahashwan SA, El Shafey HM, Fayed AA, Alhhazmi A, Manzoor N. Antimicrobial resistance pattern of MRSA strains isolated from patients of a hospital in Madinah, Kingdom of Saudi Arabia. African J Microbiol Res.2018;12(47):1044-1049.
Becker K, von Eiff C. Staphylococcus, Micrococcus and Other Catalase-positive cocci: Characterization of Colonizing S. aureus In: Versalovic J, Carroll KC, Funke G, Jorgensen JH, Landry HL, Warnock DW. Editors. manual of clinical microbiology, 10th Edition, ASM press, Washington, D.C. 2011;23-45.
Mellmann A, Weniger T, Berssenbrugge C, Keckevoet U, Friedrich AW. Characterization of clonal relatedness among the natural population of Staphylococcusaureus strains by using spa sequence typing and the BURP (Based Upon Repeat Patterns) algorithm. J ClinMicrobiol.2008;46:2805–2808.
Kolawole DO, Adeyanju A, Schaumburg F, Akinyoola AL, Lawal OO. Characterization of colonizing Staphylococcusaureus isolated from surgical wards’ patients in a Nigerian University Hospital. PLOS One. 2013;8(7):68-721.
Lowy FD. Staphylococcusaureus Infections. New Engl J Med.1998;339:520–532.
Lowy FD, Aiello AE, Bhat M, Johnson-Lawrence VD, Lee MH, Burrell E, Wright LN, Vasquez G, Larson EL. Staphylococcus aureuscolonization and infection in New York State prisons. J Infect Dis.2007;196:911–918.
Omidi M, Firoozeh F, Saffari M, Sedaghat H, Zibaei M, Khaledi A. Ability of biofilm production and molecular analysis of spa and icagenes among clinical isolates of Methicillin Resistant Staphylococcusaureus. BMC Res Notes.2020;13:19-45.
Liu Y Zhang J, Ji Y. Environmental factors modulate biofilm formation by Staphylococcusaureus. Sci Program. 2020;103:23-56.
Gaire U, Shrestha TU, Adhikari S, Adhikari N, Bastola A, Rijal KR, Ghimire P, Banjara MR. Antibiotic susceptibility, biofilm production, and detection of mecA gene among Staphylococcus aureus isolates from different clinical specimens. Diseases. 2021;9:80-109.
Oche DA, Abdulrahim U, Oheagbulem AS, Olayinka BO. Isolation of biofilm producing Methicillin-Resistant Staphylococcus aureusfrom hospitalized orthopaedic patients in Kano State, Nigeria.Niger J Basic Appl Sci. 2020;28(1):66-74.
Kirti L, Jyoti S, Pratibha M, Sumit L. Prevalence pattern of MRSA from a Rural Medical College of North India: A cause of concern. J Family Med Prim Care.2021;23:34-67.
Kot B, Wierzchowska K, Piechota M, Grużewsk A. Antimicrobial resistance patterns in Methicillin Resistant Staphylococcus aureus from patients hospitalized during 2015–2017 in Hospitals in Poland. Mediterr Princ Pract. 2020;29:61–68.
Uribe-GarcíaA, Paniagua-Contreras GL, Monroy-Pérez E, Bustos-Martínez M, Hamdan-Partida A, Garzón J, Alanís J, Quezada R, Vaca-Paniagua F, Vaca S. Frequency and expression of genes involved in adhesion and biofilm formation in Staphylococcus aureus strains isolated from periodontal lesions. J Microbiol, Immunol Infect.2021;54(2):267-275.
Alli OA, Ogbolu DO, Shittu AO, Okorie AN, Akinola JO, Daniel, JB. Association of virulence genes with mecA gene in Staphylococcusaureus isolates from Tertiary Hospitals in Nigeria. Indian J PatholMicrobiol. 2015;58(4):464–71.
Mohammadi A, Goudarzi M, Dadashi M, Soltani M, Goudarzi H, Hajikhani B. Molecular detection of genes involved in biofilm formation in Staphylococcus aureusstrains isolates: Evidence from ShahidMotahari Hospital in Tehran. Jundish J Microbiol.2020;13(7):10-2058.
Chen K, Lin S, Li P, Song Q, Luo D, Liu T, Zeng L, Zhang W. Characterization of Staphylococcusaureus isolated from patients with burns in a regional burn center, Southeastern China. BMC Infect Dis. 2018;18:51-60.
Seni J, Bwanga F, Najjuka CF, Makobore P, Okee M. Molecular characterization of Staphylococcusaureus from patients with surgical site infections at Mulago Hospital in Kampala, Uganda. PLOS One. 2013;8(6):66-153.
Mshana SE, Kamugisha E, Mirambo M, Chalya P, Rambau P. Prevalence of clindamycin inducible resistance among Methicillin-resistant Staphylococcusaureus at Bugando Medical Centre, Mwanza, Tanzania. Tanz J Health Res.2009;11:59–64.
Nwode V. Detection of Methicillin Resistant Staphylococcus aureus and associated risk factors among wound patient in a Tertiary Hospital in Abakaliki, (M. Sc Thesis at Ebonyi state University, Abakaliki; 2021.
Obajuluwa AF, Onaolapo JA, Oyi AR, Olayinka BO. Susceptibility profile of Methicillin-ResistantStaphylococcus aureus(MRSA) isolates toantibiotics and methanolic extracts of Parkiabiglobosa(Jacq) Benth. British J Pharm Res. 2013;3(4):587-596.
Onolitola OS, Olayinka BO, Salawu MJ, Yakubu SE. Nasal carriage of Methicillin Resistant Staphylococcus aureuswith Reduced Vancomycin Susceptibility (MRSA‑RVS) by Healthy Adults in Zaria. Niger J Tropical MicrobiolBiotechnol. 2007;3:19‑22.
Taiwo SS, Bamigboye TB, Odaro O, Adefioye OA, Fadiora SO. Vancomycin intermediate and high level resistant Staphylococcus aureus clinical isolates in Osogbo, Nigeria. J Microbiolog Res. 2011;3:5-22.
Chelkeba L, Melaku T. Epidemiology of Staphylococci Species and their antimicrobial-resistance among patients with wound infection in Ethiopia: A systematic review and meta-analysis. J Glob Antimicrob Resist. 2021;23:34-78.
Tania N, Shamsuzzaman SM, Islam A. Antimicrobial resistance and quorum sensing genes detection among the biofilm forming Staphylococcus aureus isolated from admitted patients of Dhaka Medical College Hospital, Dhaka, Bangladesh. Fortune J Health Sci.2021;4(3):441-455.
Shekarabi M, Hajikhani B, Salimi CA, Fazeli M, Goudarzi M. molecular characterization of vancomycin-resistant Staphylococcus aureus strains isolated from clinical samples: A Three Year Study in Tehran, Iran. PLOS One.2017;12(8):183-607.
Neopane P, Nepal HP, Shrestha R, Uehara O, Abiko Y. In vitro biofilm formation by Staphylococcus aureus isolated from wounds of hospital-admitted patients and their association with antimicrobial resistance. J Gen Internal Med.2018;11:25–32.
Azmi K, Qrei W, Abdeen Z. Screening of genes encoding adhesion factors and biofilm production in Methicillin Resistant Strains of Staphylococcus aureus isolated from palestinian patients. BMC genomics, 2019;20(1):571-578.
Abdullahi N, Iregbu KC. Methicillin-Resistant Staphylococcus aureusin a Central Nigeria Tertiary Hospital. Annal of Trop Pathol. 2019;9:6-10.
Khasawneh AI, Himsawi N, Abu-Raideh J, Salameh MA, Al-Tamimi M, Al Haj Mahmoud S, Saleh T. Status of biofilm-forming genes among jordanian nasal carriers of methicillin-sensitive and Methicillin-Resistant Staphylococcus aureus. Iranian Biomed J.2020;23:34-78.
Tahaei SAS, Stájer A, Barrak I, Ostorházi E, Szabó D, Gajdács M. Correlation between biofilm-formation and the antibiotic resistant phenotype in Staphylococcusaureus isolates: A laboratory-based study in Hungary and a review of the literature. Infect Drug Resist.2021;14:1155–1168.
Falagas ME, Karageorgopoulos DE, Leptidis J, Korbila IP. MRSA in Africa: Filling the global map of antimicrobial resistance. PLOS One. 2013;8(7):680-24.
Kong EF, Johnson JK, Jabra-Rizk MA. Community associated Methicillin-resistant Staphylococcus aureus: an enemy amidst us. PLOS Pathogens. 2016;12(10):100-5837.
Maharjan B, Karki ST, Maharjan R. Antibiotic susceptibility pattern of Staphylococcus aureusisolated from Pus/Wound swab from children attending International Friendship Children's Hospital. Nepal J Biotechnol. 2021;9(1):8-17.
Ariom TO, Iroha I R, Moses IB, Iroha CS, Ude UI, Kalu AC. Detection and phenotypic characterization of Methicillin-Resistant Staphylococcus aureus from clinical and community samples in Abakaliki, Ebonyi State, Nigeria. Afri Health Sci. 2019;19(2):2026-2035.
Gurung RR, Maharjan P, Chhetri GG. Antibiotic resistance pattern of Staphylococcusaureus with Reference to MRSA isolates from pediatric patients. Future Sci. 2020;6(4):34-67.
Abstract View: 108 times
PDF Download: 42 times