Staphylococcus aureus and Staphylococcal enterotoxin detection in raw milk and cheese origin coagulase positive isolates

Abstract

In the present study, a total 110 (60 cheese and 50 cows’ raw milk) samples was analyzed for CPS according to Food Drug Administration (FDA, 2001) previously. After the isolation, to confirmation of the isolates, catalase test, microscopic examination, coagulase test in tube and glucose-mannitol fermentation tests were applied. After the tests, we obtained 97 CPS isolates. The 97 CPS isolates were used as a material. For the confirmation of the isolates being Staphylococci species, 16S rRNA was detected by using PCR assay. For the detection of the CPS isolates being S. aureus, nuc gene detected in the CPS isolates using PCR assay. To determination of the staphylococcal enterotoxin, PCR assay was applied. The 16 S rRNA was detected in a total 97 (35 milk origin and 62 cheese origin) isolates. Therefore, these isolates were evaluated as coagulase-positive staphylococci (CPS).  The nuc gene was detected in 50 out of 97 CPS isolates. So, the 50 (18 cheese origin and 32 milk origin) isolates were evaluated as S. aureus. However, none of the staphylococcal enterotoxin genes

(SE A,B,C,D,E,G,H,I,J) was detected in 97 CPS or 50 S. aureus isolates.

Keywords

• Food Contamination,Bacterial Toxin,CPS,nuc Gene

References

1. Argudin, M.A., Mendoza, M.C., & Rodicio, M.R. 2010. Food poisoning and Staphylococcus aureus enterotoxins, Toxins, 2: 1751-1773.
2. Asao, T., Kumeda, Y., Kawai, T., Shibata, T., Oda, H., Haruki, K., Nakazawa, H., Kozaki, S. 2003. An extensive outbreak of staphylococcal food poisoning due to low-fat milk in Japan: estimation of enterotoxin A in the incriminated milk and powdered skim milk, Epidemiol, Infect, 130: 33-40.
3. Beuvier, E., Buchin, S. 2004. Raw milk cheeses. Cheese: Chemistry, Physics And Microbiology, 1, 319-345.
4. Fox, P. F., McSweeney, P.L.H., Cogan, T.M., Guinee, T.P. 2004. Cheese: Chemistry, Physics and Microbiology, Elsevier Academic Press, Amsterdam.
5. Bianchi, D.M., Gallina, S., Bellio, A., Chiesa, F., Civera, T., Decastelli, L. 2014. Enterotoxin gene profiles of Staphylococcus aureus isolated from milk and dairy products in Italy, Letters in Applied Microbiology, 58: 190-196.
6. Callon, C., Gilbert, F.B., Cremoux, R.D., Montel, M.C. 2008. Application of variable number of tandem repeat analysis to determine the origin of S. aureus contamination from milk to cheese in goat cheese farms, Food Control, 19: 143-150.
7. Carfora, V., Caprioli, A., Marri, N., Sagrafoli, D., Boselli, C., Giacinti, G., Giangolini, G., Sorbara, L., Dottarelli, S., Battisti, A., Amatiste, S. 2015. Enterotoxin genes, enterotoxin production, and methicillin resistance in Staphylococcus aureus isolated from milk and dairy products in Central Italy, International Dairy Journal, 42: 12-15.
8. Cha, J.O., Lee, J.K., Jung, Y.H., Yoo, J.I., Park, Y.K., Kim, B.S., Lee, Y.S. 2006. Molecular analysis of Staphylococcus aureus isolates associated with staphylococcal food poisoning in South Korea, Journal of Applied Microbiology, 101: 864-871.

9. Charlier, C., Even, S., Gautier, M., Le Loir, Y. 2008. Acidification is not involved in the early inhibition of Staphylococcus aureus growth by Lactobacillus lactis in milk, International Dairy Journal, 18: 197-203.
10. De Buyser, M.L., Dufour, B., Maire, M., Lafarge, V. 2001. Implication of milk and milk products in food-borne diseases in France and in different industrialized countries, International Jounal of Food Microbiology, 67: 1-17.
11. Doyle, M.P., Beuchat, L.R., Montville, T.J. 2007. Food Microbiology: Fundamentals and Frontiers, 3rd ed, American Society for Microbiology, Washington DC.
12. EFSA and ECDC. 2016. The European Union summary report on trends and sources of zoonoses, zoonotic agents and foodborne outbreaks in 2015. EFSA J, 14-4634.
13. Gallina, S., Bianchi, D.M., Bellio, A., Nogarol, C., Macori, G., Zaccaria, T., Biorci, F., Carraro, E., Decastelli, L. 2013. Staphylococcal poisoning foodborne outbreak: Epidemiological investigation and strain genotyping, Journal of Food Protection, 76: 2093-2098.
14. Gökmen, M., Gürbüz, Ü., Torlak, E., İnal, M. 2013. Identification of Staphylococcus spp. isolated in different production stages of white cheese and detection of enterotoxin, Kocatepe VeterinaryJournal, 6(2): 7-11.
15. Hummerjohann, J., Naskova, J., Baumgartner, A., & Graber, H.U. 2014. Enterotoxin producing Staphylococcus aureus genotype B as a major contaminant in Swiss raw milk cheese, Journal of Dairy Science, 97: 1305-1312.
16. Janštová, B., Necıdová, L., Skočková, A., Janštová, B. 2014. Staphylococcal enterotoxin production in model samples of milk and fresh cheese, Journal of Food and Nutrition Research, 53: 389-392.
17. Le Loir, Y., Baron, F., Gautier, M. 2003. Staphylococcus aureus and food poisoning, Genetics and Molecular Research, 2(1): 63-76.
18. McClure, J.A., Conly, J.M., Lau, V., Elsayed, S., Louie, T., Hutchins, W., Zhang, K. 2006. Novel multiplex PCR assay for detection of the staphylococcal virulence marker-Panton-Valentine leucocidin genes and simultaneous discrimination of methicillin- susceptible from resistant staphylococci, Journal of Clinical Microbiology, 44: 1141–1144.
19. Medveďová, A., Valík, L., Sirotná, Z., Liptáková, D. 2009. Growth characterisation of Staphylococcus aureus in milk: A quantitative approach, Czech Journal of Food Sciences, 27: 443–453.
20. Mendoza, M.C. 2004. Genetic procedures for identifica¬tion of enter toxigenic strains of Staphylococcus aureus from three food poisoning outbreaks, Jounal of Food Microbiology, 94(3): 279-286.
21. Necidova, L., Bogdanovicova, K., Harustiakova, D., Bartova, K. 2016. Short communication: Pasteurization as a means of inactivating staphylococcal enterotoxins A, B, and C in milk, Journal of Dairy Science, 99: 8638–8643.
22. Ono, H.K., Sato'o, Y., Narita, K., Naito, I., Hirose, S., Hisatsune, J., Asano, K., Hu, D.L., Omoe, K., Sugai, M., Nakane, A. 2015. Identification and characterization of a novel staphylococcal emetic toxin, Applied and Environmental Microbiology, 81: 7034–7040.
23. Ostyn, A., De Buyser, M., Guillier, F., Groult, J., Felix, B., Salah, S., Delmas, G., Hennekinne, J. 2010. First evidence of a food poisoning outbreak due to staphylococcal enterotoxin type E, Euro Surveill, 15(13): 1-4.
24. Pedonese, F., D’ascenzi, C., Torracca, B., Zingoni, C., Turchi, B., Fratini, F., Nuvoloni, R. 2014. Staphylococcus aureus growth and enterotoxin production in Italian caciotta cheese, Turkish Journal of Veterinary and Animal Science, 38: 318-324.
25. Presscott, L.M., Harley, J. P., Klein, D.A. 2012. Text book of Microbiology. Brown Publishers. 5th edition, 441-442.
26. Rahbar Saadat, Y., Fooladi, A.A., Shapouri, R., Hosseini, M.M., Khiabani, Z.D. 2014. Prevalence of enterotoxigenic Staphylococcus aureus in organic milk and cheese in Tabriz, Iran, Iran Journal of Microbiology, 6(5): 345-349.
27. Rall, V.L.M., Vieira, F.P., Rall, R., Vieitis, R.L., Fernandes, A. Jr., Candeias, J.M.G., Cardoso, K.F.G., Araujo, J.P. r. 2008. PCR detection of staphylococcal enterotoxin genes in S. aureus strains isolated from raw and pasteurized milk, Veterinary Microbiology, 132: 408-413.
28. Riva, A., Borghi, E., Cirasola, D., Colmegna, S., Borgo, F., Amato, E., Pontello, M.M., Morace, G. 2015. Methicillin-resistant Staphylococcus aureus in raw milk: Prevalence, SCC mec typing, enterotoxin characteri¬zation, and antimicrobial resistance patterns, Journal of Food Protection, 78(6): 1142-1146.
29. Rola, J.G., Czubkowska, A., Korpysa-Dzirva, W., Osek, J. 2016. Occurrence of Staphylococcus aureus on Farms with Small Scale Production of Raw Milk Cheeses in Poland, Toxin, 8(62): 1-9.
30. Schelin, J., Wallin-Carlquist, N., Cohn, M.T., Lindqvist, R., Barker, G.C., Rådström, P. 2011. The formation of Staphylococcus aureus enterotoxin in food environments and advances in risk assessment, Virulence, 2: 580–592.
31. Shimizu, A., Fujita, M., Igarashi, H., Takagi, M., Nagase, N., Sasaki, A., Kawano, J. 2000. Characterization of Staphylococcus aureus coagulase type VII isolates from staphylococcal food poisoning outbreaks (1980–1995) in Tokyo, Japan, by pulsed field gel electrophoresis, Journal Clinical Microbiology, 38: 3746–3749.
32. Simeão do Carmo, L., Dias, R.S., Linardi, V.R., José de Sena, M., Aparecidados Santos, D., Eduardo de Faria, M., Pena, E.C., Jett, M., Heneine, L.G. 2002. Food poisoning due to enterotoxigenic strains of Staphylococcus present in Minas cheese and raw milk in Brazil, Food Microbiology, 19(1): 9-14.
33. Tasci, F., Sahindokuyucu, F., Ozturk, D. 2011. Detection of Staphylococcus species and staphylococcal enterotoxins by ELISA in ice cream and cheese consumed in Burdur Province, African Journal of Agricultural Research, 6(4): 937-942.
34. Todd, E.C., Michaels, B.S., Greig, J.D., Smith, D., Holah, J., Bartleson, C.A. 2010. Outbreaks where food workers have been implicated in the spread of foodborne disease. Part 7. Barriers to reduce contamination of food by workers, Journal of Food Protection, 73: 1552–1565.
35. Toubar S.M.E., Elbialy, A.A., Zaky, M.M.M., El-Shafey, A.A. 2018. Prevalence of Staphylococcus aureus in Raw Milk and Some Dairy Products in Port Said Governorate, American Journal of Zoology, 1(2): 40-46.
36. Waage, S., Mørk, T., Røros, A., Aasland, D., Hunshamar, A., Odegaard, S. A. 1999. Bacteria associated with clinical mastitis in dairy heifers, Journal of Dairy Sceience, 82: 712–719.
37. Williams, A.G., Withers, S.E. 2010. Microbiological characterization of artisanal farm house cheeses manufactured in Scotland, International Journal of Dairy Technology, 63: 356–369.
38. Yücel, N., Anıl, Y., Identification and Antimicrobial Susceptibility of Staphylococcus aureus and Coagulase Negative Staphylococci Isolated from Raw Milk and Cheese Samples, Turk Hijyen ve Deneysel Biyoloji Dergisi, 68 (2): 73-78, 2011.