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Characterization of biofilms and antimicrobial resistance of coagulase-negative Staphylococcus species involved with subclinical mastitis

Published online by Cambridge University Press:  17 May 2021

Márcia Silva Francisco
Affiliation:
Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
Ciro César Rossi
Affiliation:
Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
Maria Aparecida Vasconcelos Paiva Brito
Affiliation:
Embrapa Gado de Leite, Juiz de Fora, MG, Brazil
Marinella Silva Laport
Affiliation:
Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
Elaine Menezes Barros
Affiliation:
Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
Marcia Giambiagi-deMarval*
Affiliation:
Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
*
Author for correspondence: Marcia Giambiagi-deMarval, Email: [email protected]

Abstract

Biofilm formation is a central feature to guarantee staphylococcal persistence in hosts and is associated with several diseases that are difficult to treat. In this research paper, biofilm formation and antimicrobial susceptibility were investigated in staphylococcal strains belonging to several species. These strains were isolated from the milk of cows with subclinical mastitis and most of them were coagulase-negative, with the prevalence of Staphylococcus chromogenes. High genetic diversity was observed among the strains by pulsed field gel electrophoresis. Antimicrobial resistance was assessed by disk diffusion and more than 50% of the strains were resistant to ampicillin and penicillin G, with multi-resistance profiles (13.6%) also being observed. Most strains (65.9%) formed biofilms when cultivated in BHI supplemented with 1% glucose. Most strains (72.7%) carried the intercellular adhesion gene (icaA), while less than half (36.3%) carried the biofilm-associated protein gene (bap). Concentrations of up to 10xMIC of erythromycin and tetracycline were not sufficient to suppress cell viability in preformed biofilms. Our results revealed that a genetically diverse group of biofilm-forming Staphylococcus species can be involved in subclinical mastitis. Since high antimicrobial concentrations cannot eradicate biofilm cells in vitro, their use in dairy animals may be ineffective in controlling infections, while supporting selection of resistant microorganisms. These data reinforce the need for alternative therapies aiming at disrupting biofilms for effective disease control.

Type
Research Article
Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

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Footnotes

*

These authors contributed equally to this work.

References

Aarestrup, FM (2015) The livestock reservoir for antimicrobial resistance: a personal view on changing patterns of risks, effects of interventions and the way forward. Philosophical Transactions of the Royal Society B: Biological Sciences 370, 20140085.CrossRefGoogle ScholarPubMed
Acosta, AC, Silva, LBG, Medeiros, ES, Pinheiro-Júnior, JW and Mota, RA (2016) Mastitis in ruminants in Brazil. Pesquisa Veterinária Brasileira 36, 565573.CrossRefGoogle Scholar
Ahn, SJ, Lemos, JA and Burne, RA (2005) Role of HtrA in growth and competence of Streptococcus mutans UA159. Journal of Bacteriology 187, 30283038.CrossRefGoogle ScholarPubMed
Arciola, CR, Baldassarri, L and Montanaro, L (2001) Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter-associated infections. Journal of Clinical Microbiology 39, 21512156.CrossRefGoogle Scholar
Balcão, LF, Longo, C, Costa, JHC, Uller-Gómez, C, Filho, LCPM and Hötzel, MJ (2016) Characterisation of smallholding dairy farms in southern Brazil. Animal Production Science 57, 735745.CrossRefGoogle Scholar
Becker, K, Heilmann, C and Peters, G (2014) Coagulase-negative staphylococci. Clinical Microbiology Reviews 27, 870926.Google ScholarPubMed
Busanello, M, Rossi, RS, Cassoli, LD, Pantoja, JCF and Machado, PF (2017) Estimation of prevalence and incidence of subclinical mastitis in a large population of Brazilian dairy herds. Journal of Dairy Science 100, 65456553.CrossRefGoogle Scholar
CLSI (2016) Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals, Document VET01-A4. Clinical and Laboratory Standards Institute 4th ed., Wayne, PA.Google Scholar
Coimbra-e-Souza, V, Rossi, CC, Jesus-deFreitas, LJ, Brito, MAVP, Laport, MS and Giambiagi-deMarval, M (2019) Diversity of species and transmission of antimicrobial resistance among Staphylococcus spp. isolated from goat milk. Journal of Dairy Science 102, 55185524.CrossRefGoogle ScholarPubMed
Cote-Gravel, J and Malouin, F (2019) Symposium review: features of Staphylococcus aureus mastitis pathogenesis that guide vaccine development strategies. Journal of Dairy Science 102, 47274740.CrossRefGoogle ScholarPubMed
Cucarella, C, Solano, C, Valle, J, Amorena, B, Lasa, I and Penades, JR (2001) Bap, a Staphylococcus aureus surface protein involved in biofilm formation. Journal of Bacteriology 183, 28882896.CrossRefGoogle ScholarPubMed
Flemming, K, Klingenberg, C, Cavanagh, JP, Sletteng, M, Stensen, W, Svendsen, JS and Flaegstad, T (2009) High in vitro antimicrobial activity of synthetic antimicrobial peptidomimetics against staphylococcal biofilms. Journal of Antimicrobial Chemotherapy 63, 136145.CrossRefGoogle ScholarPubMed
Flemming, HC, Wingender, J, Szewzyk, U, Steinberg, P, Rice, SA and Kjelleberg, S (2016) Biofilms: an emergent form of bacterial life. Nature Reviews Microbiology 14, 563575.CrossRefGoogle ScholarPubMed
Gomes, F, Saavedra, MJ and Henriques, M (2016) Bovine mastitis disease/pathogenicity: evidence of the potential role of microbial biofilms. Pathogens and Disease 74, ftw006.CrossRefGoogle ScholarPubMed
Madsen, JS, Burmolle, M, Hansen, LH and Sorensen, SJ (2012) The interconnection between biofilm formation and horizontal gene transfer. FEMS Immunology and Medical Microbiology 65, 183195.CrossRefGoogle ScholarPubMed
Marsilio, F, Di Francesco, CE and Di Martino, B (2018) Coagulase-positive and coagulase-negative staphylococci animal diseases. Pet-to-man travelling staphylococci: a world in progress. Academic Press 1, 4350.Google Scholar
Mesquita, AA, Rocha, CMBM, Bruhn, FRP, Custódio, DAC, Braz, MS, Pinto, SM, Silva, DB and Costa, GM (2019) Staphylococcus aureus and Streptococcus agalactiae: prevalence, resistance to antimicrobials, and their relationship with the milk quality of dairy cattle herds in Minas Gerais state, Brazil. Pesquisa Veterinária Brasileira 39, 308316.Google Scholar
Nunes, AP, Teixeira, LM, Bastos, CC, Silva, MG, Ferreira, RB, Fonseca, LS and Santos, KR (2005) Genomic characterization of oxacillin-resistant Staphylococcus epidermidis and Staphylococcus haemolyticus isolated from Brazilian medical centres. Journal of Hospital Infection 59, 1926.CrossRefGoogle ScholarPubMed
O'Gara, JP (2007) ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus. FEMS Microbiology Letters 270, 179188.CrossRefGoogle ScholarPubMed
Oliver, SP, Gonzalez, RN, Hogan, JS, Jayarao, BM and Owens, WE (2004) Microbiological Procedures for the Diagnosis of Bovine Udder Infection and Determination of Milk Quality, 4th Edn. Verona, WI, USA: National Mastitis Council.Google Scholar
Otto, M (2013) Staphylococcal infections: mechanisms of biofilm maturation and detachment as critical determinants of pathogenicity. Annual Review of Medicine 64, 175188.CrossRefGoogle ScholarPubMed
Pitcher, DG, Saunders, NA and Owen, RJ (1989) Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Letters in Applied Microbiology 8, 151156.CrossRefGoogle Scholar
Potter, A, Ceotto, H, Giambiagi-Demarval, M, dos Santos, KR, Nes, IF and Bastos Mdo, C (2009) The gene bap, involved in biofilm production, is present in Staphylococcus spp. strains from nosocomial infections. Journal of Microbiology 47, 319326.CrossRefGoogle ScholarPubMed
Rossi, CC, Pereira, MF and Giambiagi-deMarval, (2020) Underrated Staphylococcus species and their role in antimicrobial resistance spreading. Genetics and Molecular Biology 43, e20190065.CrossRefGoogle ScholarPubMed
Salina, A, Guimarães, FF, Richini Pereira, VB, Menozzi, BD, Rall, VLM and Langoni, H (2020) Detection of icaA, icaD and bap gene and biofilm production in Staphylococcus aureus and non-aureus staphylococci isolated from subclinical and clinical bovine mastitis. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 72, 10341038.CrossRefGoogle Scholar
Sambrook, J and Russell, DW (2001) Molecular Cloning: A Laboratory Manual, 3rd Edn., New York: Cold Spring Harbor Laboratory Press Cold Spring Harbor.Google Scholar
Santos, OCS, Barros, EM, Brito, MAVP, Bastos, MCF, Santos, KRN and Giambiagi-deMarval, (2008) Identification of coagulase-negative staphylococci from bovine mastitis using RFLP-PCR of the groEL gene. Veterinary Microbiology 130, 134140.CrossRefGoogle ScholarPubMed
Schwarz, S, Febler, AT, Loncaric, I, Wu, C, Kadlec, K, Wang, Y and Shen, J (2018) Antimicrobial resistance among staphylococci of animal origin. Microbiology Spectrum 6, 4.CrossRefGoogle Scholar
Stepanovic, S, Vukovic, D, Hola, V, Di Bonaventura, G, Djukic, S, Cirkovic, I and Ruzicka, F (2007) Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS 115, 891899.CrossRefGoogle ScholarPubMed
Van Belkum, A, Tassios, PT, Dijkshoorn, L, Haeggman, S, Cookson, B, Fry, NK, Fussing, V, Green, J, Feil, E, Gerner-Smidt, P, Brisse, S and Struelens, M (2007) Guidelines for the validation and application of typing methods for use in bacterial epidemiology. Clinical Microbiology and Infection 13, 146.CrossRefGoogle ScholarPubMed
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