Producción de biofilm y presencia de genes icaABCD en cepas de Staphylococcus aureus aisladas de leche cruda.

Messaria Ginestre P., Yeiny Ávila R., Kutchynskaya Valero L., Jhoandry Rivera S., Wilfrido Briñez Z., Robert Valeris C.

Resumen


En la patogénesis de la mastitis estafilocócica, el biofilm se considera un marcador de virulencia que permite la adherencia del microorganismo al epitelio mamario. Con la finalidad de determinar la producción de biofilm y detectar la presencia de genes icaABCD, se analizaron 30 cepas de S. aureus aisladas de leche cruda obtenida de vacas con mastitis subclínica. El biofilm se determinó por el método cuantitativo sobre microplacas de cultivo celular y los genes icaABCD por reacción en cadena de la polimerasa. En 96,67% de las cepas de S. aureus se observó producción de biofilm. De éstas 43,34% fueron fuertes productoras, 30,00% moderadas productoras, 23,33% débiles productoras y 3,33% no productoras de biofilm. La totalidad de los genes icaADCD se obtuvo en 6,66% de los aislamientos. Todas las cepas, excepto una, presentaron icaA o icaD. En 56,66% de los aislamientos se identificaron genes icaA/icaD. En todas las cepas productoras de biofilm se identificaron genes del operón icaABCD, lo cual evidencia que poseen un factor de virulencia que facilita su persistencia en el tejido mamario. Además, el biofilm constituye una fuente de contaminación microbiana que puede conducir a fallas en los procesos de producción que generen problemas de inocuidad de los alimentos.

Palabras clave


Biofilm; S. aureus; genes ica; leche cruda.

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Referencias


Cramton S, Gerke C, Schnell N, Nichols W, Gotz F. The intercellular adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation. Infect Immun. 1999; 67: 5427-5433.

Cucarella C, Tormo A, Ubeda C, Trotonda P, Monzon M, Peris C, et al. Role of biofilm associated protein Bap in the pathogenesis of bovine Staphylococcus aureus. Infect Immun, 2004; 72: 2177-2185.

Fox L, Zadoks R, Gaskins C. Biofilm production by Staphylococcus aureus associated with intramammary infection. Vet Microbiol. 2005; 107:295-299. 16.

Melchior M, Vaarkamp H, Fink J. Biofilms: A role in recurrent mastitis infections? Vet J. 2006; 171: 398-407.

Vasudevan P, Nair M, Annamalai T, Venkitanarayanan K. Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Vet Microbiol. 2003; 92: 179-185.

Christensen G, Simpson W, Younger J, Baddour L, Barrett F, Melton D, et al. Adherence of Coagulase-Negative Staphylococci to Plastic Tissue Culture Plates: a quantitative model for the adherence of Staphylococci to medical devices. J. Clin Microbiol 1985; 22: 996-1006.

Aguiar F, Medeiros F, Fernandes O, Gudziki R, Perdreau F, Riley L. New Staphylococcus aureus genotyping method based on exotoxin (set) genes. J Clin Microbiol. 2004; 44: 2728-2732.

Greisen K, Loeffelholz M, Purohit A, Leong D. PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid. J. Clin Microbiol. 1994; 32: 335-351.

Ramesh A, Padmapriya B, Chrashekar A. Application of a convenient DNA extraction method and multiplex PCR for the direct detection of Staphylococcus aureus and Yersinia enterocolitica in milk samples. Mol Cell Probe. 2002; 16: 307314.

Kiem S, Oh W, Peck K, Lee N, Lee J, Song J, et al. Phase variation of biofilm formation in Staphylococcus aureus by IS256 insertion and its impact on the capacity adhering to polyurethane surface. J Korean Med Sci. 2004; 19: 779-782.

Gad G, El-Feky M, El-Rehewy M, Hassan M, Abolella H, El-Baky R. Detecton of icaA, icaD and bioflim production by Staphylococcus aureus and Staphylococcus epidermidis isolated from urinary tract catheterized patients. J Infect Dev Ctries. 2009; 3(5): 342-351.

Ausubel F, Brent R, Kingston, R, Moore, D, Seidam J, Smith J, Struhl K.. Short protocols in molecular biology: a compendium of methods from current protocols in molecular biology. 5a edition, Vol. 2. John Wiley & Sons, Inc. USA. 2002. 1512 p.

De los Santos R, Fernández M, Carro S, Zunino P. Characterisation of Staphylococcus aureus isolated from cases of bovine subclinical mastitis in two Uruguayan dairy farms. Arch Med Vet. 2014; 46(2): 315-320.

Peña J, Uffo O. Producción de biofilm en genotipos de Staphylococcus aureus aislados de mastitis bovina en Cuba. Rev Salud Anim 2013; 35(3): 189196.

De Castro P, Ferreira L, Filho A, Zafalon L, Vicente H, de Souza V. Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis. Braz. J Microbiol. 2013; 44(1): 119-124.

Pereyra E, Picech F, Renna M, Baravalle C, Andreotti C, Russi R, et al. Detection of Staphylococcus aureus adhesion and biofilm-producing genes and their expression during internalization in bovine mammary epithelial cells. Vet Microbiol. 2016; 1: 83:69-77.

Milanov D, Laziæ S, Vidiae B, Petroviae J, Bugarski D, Šeguljev Z. Slime production and biofilm forming ability by Staphylococcus aureus bovine mastitis isolates. Acta Vet. 2010; 60(2-3):217-226.

Darwish S. Asfour H. Investigation of Biofilm Forming ability in Staphylococci causing bovine mastitis using phenotypic and genotypic assays. Scientific World J. 2103; 1-9.

Bardiau M, Detilleux J, Farnir F, Mainil J, Ote I. Associations between properties linked with persistence in a collection of Staphylococcus aureus isolates from bovine mastitis. Vet Microbiol. 2013; 12: 1-18.

Al-Rubaye S, Al-Jumaily E, Abdul-Ratha H. Biofilm production by Staphylococcus aureus isolated from bovine mastitis related with resistance to the antibiotics. Int.J.Curr.Microbiol.App.Sci. 2016; 5(5): 33-44.

Al-Iedani A. Phenotypic study on the capacity of biofilm production in Staphylococcus aureus isolated from bovine subclinical mastitis and their impact on resistance to antimicrobials. Bas J Vet Res. 2016; 15(2): 11-127.

Oliveira M, Bexiga R, Nunes S, Carneiro C, Cavaco L, Bernardo F, Vilela C. Biofilm-forming ability profiling of Staphylococcus aureus and Staphylococcus epidermidis mastitis isolates. Vet Microbiol. 2006; 118(1-2): 133-140.

Dhanawade N, Kalorey D, Srinivasan R, Barbuddhe S, Kurkure N. Detection of intercellular adhesion genes and biofilm production in Staphylococcus aureus isolated from bovine subclinical mastitis. Vet. Res. Commun. 2010; 34: 81-89.

Szweda P, Schielmann M, Milewski S, Frankowska A, Jakubczak A. Biofilm production and presence of ica and bap genes in Staphylococcus aureus strains isolated from cows with mastitis in the Eastern Poland. Pol J Microbiol. 2012; 61(1): 65-69.

Begum H, Uddin M, Islam M, Nazir K, Islam M, Rahman M. Detection of biofilm producing coagulase positive Staphylococcus aureus from bovine mastitis, their pigment production, hemolytic activity and antibiotic sensitivity pattern. J. Bangladesh Soc. Agric. Sci. Technol. 2007; 4: 97-100.

Melo P, Menezes L, Nader A, Zafalon L, Godoy H, De Souza .Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis. Braz J Microbiol. 2013; 44(1): 119-124.

Marques V, De Souza M, Mendonça E, Abreu T, Rocha B, Coelho S, et al. Análise fenotípica e genotípica da virulência de Staphylococcus spp. e de sua dispersão clonal como contribuição

ao estudo da mastite bovina. Pesq Vet Bras 2013; 33(2): 161-170.

Marques V, Motta C, Soares B, Melo D, Coelho S, Coelho I et al. Biofilm production and beta-lactamic resistance in Brazilian Staphylococcus aureus isolates from bovine mastitis. Braz J Microbiol. 2017; 48(1): 118-124

Coelho S, Pereira I, Soares L, Pribul B, Souza M. Short communication: Profile of virulence factors of Staphylococcus aureus isolated from subclinical bovine mastitis in the state of Rio de Janeiro, Brazil. J. Dairy Sci. 2011; 94(7): 3305-3310.

O’Gara J, Humphreys H. (2001) Staphylococcus epidermidis biofilms importance and implications. J Med Microbiol. 2001; 50: 582-587.




Universidad del Zulia / Venezuela / Kasmera / revistakasmera@hotmail.com / ISSN 00755222

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