Loading...

Indian Journal of Animal Research

Detection of staphylococcal enterotoxin genes in raw milk samples by realtime PCR

DOI: 10.18805/ijar.B-1067    | Article Id: B-1067 | Page : 1504-1508
Citation :- Detection of staphylococcal enterotoxin genes in raw milk samples by realtimePCR.Indian Journal of Animal Research.2019.(53):1504-1508
Akın Yiğin, Mehmet Demirci, Serap Kılıç Altun and Hikmet Dinç hikmetdnc@gmail.com
Address :
Harran University Veterinary Faculty, Department
of Pharmacology and Toxicology, Şanlıurfa, Turkey.
Submitted Date : 19-11-2018
Accepted Date : 8-03-2019
First Online:

Abstract

Presence of significant level of enterotoxigenic S. aureus in raw milk of sheep, goat and donkey may cause serious food borne disease. Many people worldwide, use raw milk in their daily life but data on presence of virulence genes in cow, sheep, goat and especially donkey milk seems to be very limited. For this reason, aim of this study was to determine the presence both S. aureus and nine staphylococcal enterotoxin genes in cow, sheep, goat and donkey milks. A total of 231 raw milk samples were collected from 48 cow, 65 goat, 65 sheep and 53 donkey were collected. To detect presence of S. aureus both conventional culture and real-time PCR were used and to detect nine staphylococcal enterotoxin genes directly in milk, real-time PCR was performed. Conventional culture and real-time PCR results were found to be similar for presence of S. aureus and it was detected in 52 (22.51%) out of 231 raw milk samples. Staphylococcal enterotoxin genes were detected in 27 out of 52 S.aureus positive samples and a total of 62 enterotoxin genes were detected in these samples. However enterotoxin genes could not be detected in two S. aureus positive donkey milk. Hence, real-time PCR proved to be reliable and faster than conventional methods to detect presence of enterotoxigenic S. aureus in milk. Raw milk samples from different animals many contain enterotoxigenic S. aureus. Therefore, one should be careful during raw milk consumption as enterotoxigenic S. aureus in milk may cause dangerous public health problem which need routine screening for this pathogen in milk.

Keywords

Raw milk Real-time PCR Staphylococcal enterotoxins.

References

  1. Carfora, V., Caprioli, A., Marri, N., Sagrafoli, D., Boselli, C., Giacinti, G., Giangolini, G., Sorbara, L., et al. (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. https://doi.org/10 .1016 j. idairyj.2014.10.009
  2. Chiefari, A.K., Perry, M.J., Kelly-Cirino K., Egan C.T. (2015). Detection of Staphylococcus aureus enterotoxin production genes from patient samples using an automated extraction platform and multiplex real-time PCR. Mol Cell Probes, 29:461-467.
  3. D’amico, D.J., Donnelly, C.W. (2011). Characterization of Staphylococcus aureus strains isolated from raw milk utilized in small-scale artisan cheese production. J Food Prot, 74:1353-8. https://doi.org/10.4315/0362-028X.JFP-10-533. 
  4. EFSA and ECDC (European Food Safety Authority and European Centre for Disease Prevention and Control).(2015). The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2013. EFSA J. 13:3991.
  5. Even, S., Charlier, C., Nouaille, S., Ben Zakour, N.L., Cretenet, M., Cousin, F.J., Gautier, M., et al. (2009). Staphylococcus aureus virulence expression is impaired by Lactococcus lactis in mixed cultures. Appl Environ Microbiol. 75:4459-72. https://    doi.org/10.1128/AEM.02388-08.
  6. Garedew. L., Berhanu, A., Mengesha, D., Tsegay, G. (2012). Identification of gram-negative bacteria from critical control points of raw and pasteurized cow milk consumed at Gondar town and its suburbs, Ethiopia. BMC Public Health. 12:950. https://doi.org/    10.1186/1471-2458-12-950.
  7. Gutiérrez, D., Delgado, S., Vázquez-Sánchez, D., Martínez, B., Cabo, M.L., Rodríguez, A., Herrera, J.J, García, P. (2012). Incidence of Staphylococcus aureus and analysis of associated bacterial communities on food industry surfaces. Appl Environ Microbiol.    78(24):8547-54. https://doi.org/10.1128/AEM.02045-12.
  8. Hein, I., Jørgensen, H.J., Loncarevic, S., Wagner, M. (2005). Quantification of Staphylococcus aureus in unpasteurised bovine and caprine milk by real-time PCR. Res Microbiol.156:554-63.https://doi.org/10.1016/j.resmic.2005.01.003.
  9. Hennekinne, J.A., De Buyser, M.L., Dragacci S. (2012). Staphylococcus aureus and its food poisoning toxins: characterization and outbreak investigation. FEMS Microbiol Rev. 36:815-36. https://doi.org/10.1111/j.1574-6976.2011.00311.x.
  10. 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. J Dairy Sci.97:1305-12. https://doi.org/10.3168/jds.2013-7643.
  11. Hunt, K., Schelin, J., Rådström, P., Butler, F., Jordan, K. (2012). Classical enterotoxins of coagulase-positive Staphylococcus aureus isolates from raw milk and products for raw milk cheese production in Ireland. Dairy Sci. & Technol. 92:487. https://doi.org/    10.1007/s13594-012-0067-4.
  12. Jakobsen, R.A., Heggebø, R., Sunde, E.B., Skjervheim, M. (2011). Staphylococcus aureus and Listeria monocytogenes in Norwegian raw milk cheese production. Food Microbiol. 28:492-6. https://doi.org/10.1016/j.fm.2010.10.017.
  13. Jørgensen, H.J., Mørk, T., Høgåsen, H.R., Rørvik, L.M. (2005). Enterotoxigenic Staphylococcus aureus in bulk milk in Norway. J Appl Microbiol. 99:158-66. https://doi.org/10.1111/j.1365-2672.2005.02569.x
  14. Kadariya, J., Smith, T.C., Thapaliya, D. (2014). Staphylococcus aureus and Staphylococcal Food-Borne Disease: An Ongoing Challenge in Public Health. Biomed Res Int. 827965. https://doi.org/10.1155/2014/827965.
  15. Klotz, M., Opper, S., Heeg, K., Zimmermann, S. (2003). Detection of Staphylococcus aureus Enterotoxins A to D by Real-Time Fluorescence PCR Assay. J Clin Microbiol. 41:4683-7.
  16. McDonald, R.R., Antonishyn, N.A., Hansen, T. ( 2005). Development of a Triplex Real-Time PCR Assay for Detection of Panton-    Valentine Leukocidin Toxin Genes in Clinical Isolates of Methicillin-Resistant Staphylococcus aureus. J Clin Microbiol.    43:6147-6149. https://doi.org/10.1128/JCM.43.12.6147-6149.2005.
  17. Mehli. L., Hoel, S., Thomassen, G.M.B., Jakobsen, A.N., Karlsen, H. (2017). The prevalence, genetic diversity and antibiotic resistance of Staphylococcus aureus in milk, whey, and cheese from artisan farm dairies. International Dairy Journal, 65, 20-27. 
  18. Merz, A., Stephan, R., Johler., S. (2016). Staphylococcus aureus Isolates from Goat and Sheep Milk Seem to Be Closely Related and Differ from Isolates Detected from Bovine Milk. Front Microbiol.14:319. https://doi.org/10.3389/fmicb.2016.00319.
  19. Morandi, S., Brasca, M., Lodi,R., Cremonesi, P., Castiglioni., B. (2007). Detection of classical enterotoxins and identification of enterotoxin genes in Staphylococcus aureus from milk and dairy products. Vet Microbiol. 20;124:66-72.
  20. Muehlherr., J.E., Zweifel, C., Corti, S., Blanco, J.E., Stephan, R. (2003). Microbiological quality of raw goat’s and ewe’s bulk-tank milk in Switzerland. J Dairy Sci. 86:3849-56.
  21. Nakayama, A., Okayama, A., Hashida, M., Yamamoto, Y., Takebe,H., Ohnaka, T., Tanaka, T., Imai, S. (2006). Development of a routine laboratory direct detection system of staphylococcal enterotoxin genes. J Med Microbiol. 55:273-7.
  22. Rola, J.G., Czubkowska, A., Korpysa-Dzirba, W., Osek, J. (2016). Occurrence of Staphylococcus aureus on Farms with Small Scale Production of Raw Milk Cheeses in Poland. Toxins (Basel).2:E62. https://doi.org/10.3390/toxins8030062.
  23. Rola, J.G., Korpysa-Dzirba, W., Czubkowska, A., Osek, J. (2015). Prevalence of enterotoxin genes and antimicrobial resistance of coagulase-positive staphylococci recovered from raw cow milk. J Dairy Sci. 98:4273-8. https://doi.org/10.3168/jds.2014-    9064.
  24. Rosengren, A., Fabricius, A., Guss, B., Sylvén, S., Lindqvist, R.(2010). Occurrence of foodborne pathogens and characterization of Staphylococcus aureus in cheese produced on farm-dairies. Int J Food Microbiol. 15;144:263-9. https://doi.org/10.1016/    j.ijfoodmicro.2010.10.004.
  25. Schelin, J., Wallin-Carlquist, N., Thorup Cohn, M., 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. https://doi.org/10.4161/    viru.2.6.18122.
  26. Walcher, G., Gonano, M., Kümmel, J., Barker, G.C., Lebl, K., Bereuter, O., Ehling-Schulz, M., et al. (2014). Staphylococcus aureus reservoirs during traditional Austrian raw milk cheese production. J Dairy Res. 81:462-70. https://doi.org/10.1017/S0022 029914000417.
  27. Xing, X., Zhang, Y., Wu, Q., Wang, X., Ge, W and Wu, C. (2016). Prevalence and characterization of Staphylococcus aureus isolated from goat milk powder processing plants Food Control, 59:644–650. https://doi.org/10.1016/j.foodcont.2015.06.042. 

Global Footprints