Virulence Factors and Biofilm Formation in Vancomycin Resistant <i>Enterococcus faecalis</i> and <i>Enterococcus faecium</i> Isolates in Brazil — Oak Academic Publishing
Research ArticleOpen AccessGoogle Scholar indexed
Virulence Factors and Biofilm Formation in Vancomycin Resistant <i>Enterococcus faecalis</i> and <i>Enterococcus faecium</i> Isolates in Brazil
Universidade Federal de Pernambuco, Pernambuco, Brazil
,
Universidade Federal de Pernambuco, Pernambuco, Brazil
,
Universidade Federal de Pernambuco, Pernambuco, Brazil
,
Universidade Federal de Pernambuco, Pernambuco, Brazil
,
Universidade Federal de Pernambuco, Pernambuco, Brazil
,
Universidade Regional do Cariri, Ceará, Brazil
,
Universidade Federal de Pernambuco, Pernambuco, Brazil
,
Instituto de Tecnologia de Pernambuco, Pernambuco, Brazil
,
Universidade Federal de Pernambuco, Pernambuco, Brazil
,
Universidade Federal de Pernambuco, Pernambuco, Brazil
1 Universidade Federal de Pernambuco, Pernambuco, Brazil
2 Universidade Federal de Pernambuco, Pernambuco, Brazil
3 Universidade Federal de Pernambuco, Pernambuco, Brazil
4 Universidade Federal de Pernambuco, Pernambuco, Brazil
5 Universidade Federal de Pernambuco, Pernambuco, Brazil
6 Universidade Regional do Cariri, Ceará, Brazil
7 Universidade Federal de Pernambuco, Pernambuco, Brazil
8 Instituto de Tecnologia de Pernambuco, Pernambuco, Brazil
9 Universidade Federal de Pernambuco, Pernambuco, Brazil
10 Universidade Federal de Pernambuco, Pernambuco, Brazil
In this work, we evaluated biofilm formation of Vancomycin Resistant of E. faecalis and E. faecium (VRE) in different culture media and adhesion substrate, as well as cellular hydrophobicity and presence of virulence genes. For this, 35 isolates were collected from a public hospital in Recife, Pernambuco, Brazil and identified by the Matrix-Assisted Laser Desorption Ionization - Time-of-flight - Mass Spectrometry (MALDI-TOF-MS) technique. Biofilm formation was analyzed by the Crystal Violet (CV) method and fluorescence microscopy, cellular hydrophobicity by hydrocarbon interaction and the presence of gel E, esp and asa 1 genes by Polymerase Chain Reaction (PCR). 12 isolates were identified as E. faecalis and 23 as E. faecium . Most were obtained in Coronary Units (40.0%) and Intensive Care Unit (31.4%). E. faecium isolates were more resistant to the antibiotics tested than E. faecalis ; however, E. faecalis stood out as a biofilm producer. Regarding the presence and gene frequency, it was observed that gel E (54.3%) and esp (54.3%) were the most prevalent, followed by asa 1 (22.9%). When comparing the gene frequency, it was observed that gel E and esp were predominant (48.6% for both species), while asa 1 was more frequent in E. faecalis (20.0%). The data presented here are worrying, because they reveal the virulence potential of isolates VRE, which contributes to the dissemination and persistence of these pathogens in the hospital environment.
Zhong, Z., Zhang, W.Y., Song, Y.Q., Liu, W.J., Xu, H.Y., Xi, X.X., et al. (2017) Comparative Genomic Analysis of the Genus Enterococcus. Microbiological Research, 196, 95-105. https://doi.org/10.1016/j.micres.2016.12.009
Amarnania, R. and Rapose, A. (2017) Colon Cancer and Enterococcus Bacteremia Coaffection: A Dangerous Aliance. Journal of Infection and Public Health, 10, 681-684. https://doi.org/10.1016/j.jiph.2016.09.009
Martínez, L.C., Álvare, C.E.G., Álvarez, M.O., Fuente del Río, R.L., Velasco, C.G. and Enciso, B.S. (2018) Meningitis Neonatal Por Enterococcus faecalis. Revista del Laboratorio Clínico, 11, 101-103. https://doi.org/10.1016/j.labcli.2017.11.007
Olmos, C., Vilacosta, I., Fernández-Pérez, C., Bernal, J.L., Ferrera, C., García-Arribas, D., et al. (2017) The Evolving Nature of Infective Endocarditis in Spain: A Population-Based Study (2003 to 2014). Journal of the American College of Cardiology, 70, 2795-2804. https://doi.org/10.1016/j.jacc.2017.10.005
Shah, K.J., Cherabuddi, K., Shultz, J., Borgert, S., Ramphal, R. and Klinker, K.P. (2018) Ampicillin for the Treatment of Complicated Urinary Tract Infections Caused by Vancomycin-Resistant Enterococcus spp (VRE): A Single-Center University Hospital Experience. International Journal of Antimicrobial Agents, 51, 57-61. https://doi.org/10.1016/j.ijantimicag.2017.06.008
Gilmore, M.S., Lebreton, F. and van Schaik, W. (2013) Genomic Transition of Enterococci from Gut Commensals to Leading Causes of Multidrug-Resistant Hospital Infection in the Antibiotic Era. Current Opinion in Microbiology, 16, 10-16. https://doi.org/10.1016/j.mib.2013.01.006
Armin, S., Fallah, F., Karimi, A., Rashidan, M., Shirdust, M. and Azimi, L. (2017) Genotyping, Antimicrobial Resistance and Virulence Factor Gene Profiles of Vancomycin Resistance Enterococcus faecalis Isolated from Blood Culture. Microbial Pathogenesis, 109, 300-304. https://doi.org/10.1016/j.micpath.2017.05.039
Uttley, A.H., Collins, C.H., Naidoo, J. and George, R.C. (1988) Vancomycin-Resistant Enterococci. The Lancet, 331, 57-58. https://doi.org/10.1016/S0140-6736(88)91037-9
Diaz-Granados, C.A., Zimmer, S.M., Klein, M. and Jernigan, J.A. (2005) Comparison of Mortality Associated with Vancomycin-Resistant and Vancomycin-Susceptible Enterococcal Bloodstream Infections: A Meta-Analysis. Clinical Infectious Diseases, 41, 327-333. https://doi.org/10.1086/430909
World Health Organization (WHO) (2017) Prioritization of Pathogens to Guide Discovery, Research and Development of New Antibiotics for Drug-Resistant Bacterial Infections, Including Tuberculosis. Geneva.
Centers for Disease Control and Prevention (CDCP) (2013) Antibiotic Resistance Threats in the United States, 2013. Atlanta.
Flemming, H.C. and Wingender, J. (2010) The Biofilm Matrix. Nature Reviews Microbiology, 8, 623-633. https://doi.org/10.1038/nrmicro2415
Trentin, D.S., Giordani, R.B. and Macedo, A.J. (2013) Biofilmes bacterianos patogênicos: Aspectos geais, importância clínica e estratégias de combate. Revista, 14, 113-238. https://doi.org/10.31514/rliberato.2013v14n22.p213
Jett, B.D., Huycke, M.M. and Gilmore, M.S. (1994) Virulence of Enterococci. Clinical Microbiology Reviews, 7, 462-478. https://doi.org/10.1128/CMR.7.4.462
Soares, R.O., Fedi, A.C., Reiter, K.C., Caierão, J. and d’Azevedo, P.A. (2014) Correlation between Biofilm Formation and gelE, Esp, and Agg Genes in Enterococcus spp. Clinical Isolates. Virulence, 5, 634-637. https://doi.org/10.4161/viru.28998
Gulhan, T., Boynukara, B., Ciftci, A., Sogut, M.U. and Findik, A. (2015) Characterization of Enterococcus faecalis Isolates Originating from Different Sources for Their Virulence Factors and Genes, Antibiotic Resistance Patterns, Genotypes and Biofilm Production. Iranian Journal of Veterinary Research, 16, 261-266.
Shankar, N., Baghdayan, A.S. and Gilmore, M.S. (2002) Modulation of Virulence within a Pathogenicity Island in Vancomycin-Resistant Enterococcus faecalis. Nature, 417, 746-750. https://doi.org/10.1038/nature00802
Upadhyaya, P.M.G., Ravikumar, K.L. and Umapathy, B.L. (2009) Review of Virulence Factors of Enterococcus: An Emerging Nosocomial Pathogen. Indian Journal of Medical Microbiology, 27, 301-305. https://doi.org/10.4103/0255-0857.55437
Clinical and Laboratory Standards Institute (CLSI) (2018) Performance Standards for Antimicrobial Susceptibility Testing, 28th Edition. CLSI supplement M100, Wayne.
Tendolkar, P.M., Baghdayan, A.S., Gilmore, M.S. and Shankar, N. (2004) Enterococcal Surface Protein, Esp, Enhances Biofilm Formation by Enterococcus faecalis. Infection and Immunity, 72, 6032-6039. https://doi.org/10.1128/IAI.72.10.6032-6039.2004
Stepanović, S., Vuković, D., Hola, V., Di Bonaventura, G., Djukić, S., Cirković, I., et al. (2007) Quantification of Biofilm in Microtiter Plates: Overview of Testing Conditions and Practical Recommendations for Assessment of Biofilm Production by Staphylococci. APMIS, 115, 891-899. https://doi.org/10.1111/j.1600-0463.2007.apm_630.x
Duprè, I., Zanetti, S., Schito, A.M., Fadda, G. and Sechi, L.A. (2003) Incidence of Virulence Determinants in Clinical Enterococcus faecium and Enterococcus faecalis Isolates Collected in Sardinia (Italy). Journal of Medical Microbiology, 52, 491-498. https://doi.org/10.1099/jmm.0.05038-0
Agência Nacional de Vigilancia Sanitária (ANVISA) (2019) Controle de infecção hospitalar: Balanço e reflexões. Ascom/Anvisa.
Cattoir, V. and Giard, J.G. (2014) Antibiotic Resistance in Enterococcus faecium Clinical Isolates. Expert Review of Anti-Infective Therapy, 12, 239-248. https://doi.org/10.1586/14787210.2014.870886
European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2018) Breakpoint Tables for Interpretation of MICs and Zone Diameters.
Agência Nacional de Vigilância Sanitária (ANVISA) (2004) Procedimentos Laboratoriais: Requisição do Exame—Análise Microbiológica. Manual de Microbiologia Clínica para o Controle de Infecção em Serviços de Saúde.
Strateva, T., Atanasova, D., Savov, E., Petrova, G. and Mitov, I. (2016) Incidence of Virulence Determinants in Clinical Enterococcus faecalis and Enterococcus faecium Isolates Collected in Bulgaria. The Brazilian Journal of Infectious Diseases, 20, 127-133. https://doi.org/10.1016/j.bjid.2015.11.011
Lima, A.V.A., Silva, S.M., Nascimento Júnior, J.A.A., Correia, M.D.S., Luz, A.C., Leal-Balbino, T.C., et al. (2020) Occurrence and Diversity of Intra- and Interhospital Drug-Resistant and Biofilm-Forming Acinetobacter baumannii and Pseudomonas aeruginosa. Microbial Drug Resistance, 26, 802-814. https://doi.org/10.1089/mdr.2019.0214
Araújo, B.C., Melo, R.C., Bortoli, M.C., Bonfim, J.R.A. and Toma, T.S. (2022) Prevention and Control of Antimicrobial Resistance in Primary Health Care: Evidence for Policies. Ciência & Saúde Coletiva, 27, 299-314. https://doi.org/10.1590/1413-81232022271.22202020
Pan American Health Organization (PAHO) (2019) Biennial Meeting of the Latin American and the Caribbean Network for Antimicrobial Resistance Surveillance. Brazil.
Oancea, C., Klare, I., Witte, W. and Werner, G. (2004) Conjugative Transfer of the Virulence Gene, Esp, among Isolates of Enterococcus faecium and Enterococcus faecalis. Journal of Antimicrobial Chemotherapy, 54, 232-235. https://doi.org/10.1093/jac/dkh249
Medeiros, A.W., Pereira, R.I., Oliveira, D.V., Martins, P.D., d’Azevedo, P.A., Van der Sand, S., et al. (2014) Molecular Detection of Virulence Factors among Food and Clinical Enterococcus faecalis Strains in South Brazil. Brazilian Journal of Microbiology, 45, 327-332. https://doi.org/10.1590/S1517-83822014005000031
Di Rosa, R., Creti, R., Venditti, M., D’Amelio, R., Arciola, C.R., Montanaro, L., et al. (2006) Relationship between Biofilm Formation, the Enterococcal Surface Protein (Esp) and Gelatinase in Clinical Isolates of Enterococcus faecalis and Enterococcus faecium. FEMS Microbiology Letters, 256, 145-150. https://doi.org/10.1111/j.1574-6968.2006.00112.x
Feng, J., Wang, T., Zhang, S., Shi, W. and Zhang, Y. (2014) An Optimized SYBR Green I/PI Assay for Rapid Viability Assessment and Antibiotic Susceptibility Testing for Borrelia burgdorferi. PLOS ONE, 9, e111809. https://doi.org/10.1371/journal.pone.0111809
Soler-Arango, J., Figoli, C., Muraca, G., Bosch, A. and Brelles-Mariño, G. (2019) The Pseudomonas aeruginosa Biofilm Matrix and Cells Are Drastically Impacted by Gas Discharge Plasma Treatment: A Comprehensive Model Explaining Plasma-Mediated Biofilm Eradication. PLOS ONE, 14, e0216817. https://doi.org/10.1371/journal.pone.0216817
Silva, S.M., Ramos, B.A., Sá, R.A.Q.C., Silva, M.V.D., Correia, M.T.S. and Oliveira, M.B.M. (2022) Investigation of Factors Related to Biofilm Formation in Providencia stuartii. Anais da Academia Brasileira de Ciências, 94, e20210765. https://doi.org/10.1590/0001-3765202220210765
Flemming, H.C., Wingender, J., Szewzyk, U., Steinberg, P., Rice, S.A. and Kjelleberg, S. (2016) Biofilms: An Emergent Form of Bacterial Life. Nature Reviews Microbiology, 14, 563-575. https://doi.org/10.1038/nrmicro.2016.94
Maillard, J.Y. and McBain, A. (2019) Biofilm in Healthcare Settings and Their Control. Letters in Applied Microbiology, 68, 268. https://doi.org/10.1111/lam.13147