Distribution of genes encoding biofilm production in S. aureus isolated from raw milk in Kurdistan

Subject Areas : Food Science and Technology

مهسا Shojaei ¹ , هیوا Karimi Darehabi ^{2 *} , جوادی javadi ³

1 - M.Sc Student in food science and technology, Science and Research Branch, Islamic Azad University, Sanandaj, Iran.
2 - Assistant Professor of Food Hygiene Department, ‍College of Veterinary Science, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran
3 - Associate Professor of Food Hygiene Department, ‍College of Veterinary Science, Tabriz Branch, Islamic Azad University, Tabriz, Iran

Received: 2013-11-09 Accepted : 2014-08-29 Published : 2014-08-23

Keywords: Staphylococcus aureus, raw milk, biofilms, Sanandaj,

Abstract :

   Staphylococcus aureus is a major cause of bovine mastitis. Several virulence factors are involved in mastitis. One of the important virulence factors is the ability of the bacterium to produce biofilms. These bacteria are capable of producing polysaccharides and proteinaceous substances attached to the surfaces, leads to biofilm formation. In this study, a total of 120 raw milk samples was obtained from Kurdistan province and analyzed for the presence of S. aureus. The presence of S. aureus was assessed by conventional culture method and confirmed by PCR assay. For this, nuc gene was exploited as the specific target sequence to detect S. aureus.  Moreover, Multiplex PCR was used to identify the presence of clfaB, fnbA, icaD and icaA genes which encode biofilm formation. Based on results, S. aureus was found in 49 (40.83%) of the samples. The frequency of each of the genes was determined as 69.38%, 32.6%, 38.77%, and 59.18 for fnbA, clfaB, icaD and icaA, respectively. Results revealed a high contamination rate of raw milks with S. aureus, and the ability of the isolates to form biofilms.

References:

• Aarnisalo, K., Lundén, J., Korkeala, H. and Wirtanen, G. (2007). Susceptibility of Listeriamonocytogenes strains to disinfectants and chlorinated alkaline cleaners at cold temperatures. LWT - Food Science and Technology, 40(6): 1041-1048.


• Anonymous. (2011). DQA—dairy quality assurance scheme regulations for Belgium. (Lastenboek IKM Productie) Version 5-11-86.


• Brakstad O.G., Aasbakk, K. and Maeland, J.A. (1992). Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. Journal. Clinical Microbiology, 30: 1654–1660.


• Ciftci, A., Findik A., Emek Onuk E. and Savasan S. (2009). Detection of meticillin resistance and slime factor production of s.aureus in bovin mastitis. Brazilian Journal of Microbiology, 40: 254-261.


• Costerton, J.W., Stewart, P.S. and Greenberg, E.P. (1999). Bacterial biofilms: a common cause of persistent infections, 284: 1318–1322.


• Duran, N., Dogramaci, Y., Ozer, B., Demir, C. and Kalaci, A. (2010). Detection of adhesin genes and slime production among Staphylococci in orthopaedic surgical wounds. African Journal of Microbiology Research, 4(9): 708-715.


• Dziewanowska, K., Patti, J.M., Deobald, C.F., Bayles, K.W., Trumble, W.R. and Bohach, G.A. (1999). Fibronectin binding protein and host cell tyrosine kinase are required for internalization of Staphylococcus aureus by epithelial cells. Infection and Immunity, 67: 4673-78.


• Foster, T.J. and Hook, M. (1998). Surface protein adhesins of Staphylococcus aureus. Journal. Trends Microbial, 6(12): 484–488.


• Gotz, F. (2002). Staphylococcus and biofilms Molecular Microbiology, (43): 1367-1378.


• Iorio, N.L., Lopes, A.P., Schuenck, R.P., Barcellos, A.G., Olendzki, A.N., Lopez, G.L., et al. (2011). A combination of methods to evaluate biofilm production may help to determine the clinical relevance of Staphylococcus in blood cultures. Microbiology and Immunology, 55(1): 28-33.


• Ní Eidhin, D., Perkins, S., Francois, P., Vaudaux, P., Hook, M., and Foster, T.J. (1998) Clumping factor B (ClfB), a new surface-located fibrinogen-binding adhesin of Staphylococcus aureus. Mol Microbiol30:245–257.


• O'Connell, D.P., Nanavaty, T., McDevitt, D., Gurusiddappa, S., Hook, M. and Foster, T.J. (1998). The fibrinogen-binding MSCRAMM (clumping factor) of Staphylococcus aureus has a Ca²⁺-dependent inhibitory site. The Journal of Biological Chemistry, 273(12): 6821–6829.


• Tarek, Z., Bochra, K., Hanene, M., Kacem, M. and Amina, B. (2010). A Microtiter plateassay for Staphylococcus aureus biofilm quantification at various pH levels and hydrogenperoxide supplementation. The New Microbiologica, 33(2): 137-145.


• Vasudevan, P., Nair, M.K., Annamalai, T. and Venkitanarayanan, K.S. (2003). Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Veterinary Microbiology, 92(1-2): 179-185.