مطالعه میزان شیوع ژن‌های آنتروتوکسین‌های معمول در استافیلوکوکوس آرئوس‌های جدا شده از شیر گاومیش‌های شهرستان تبریز به روش Multiplex PCR

محورهای موضوعی : علوم و صنایع غذایی

مهرداد اثنی عشری ¹ , جلال شایق ² , آیت الله نصرالهی عمران ³

1 - دانشگاه آزاد اسلامی، واحد تنکابن، دانشجوی کارشناسی ارشد میکروب شناسی، تنکابن، ایران
2 - دانشگاه آزاد اسلامی، واحد شبستر، استادیار گروه دامپزشکی، شبستر، ایران
3 - دانشگاه آزاد اسلامی، واحد تنکابن، دانشکده علوم زیستی، استادیار گروه میکروبیولوژی، تنکابن، ایران

تاریخ دریافت : 1391/07/26 تاریخ پذیرش : 1391/11/24 تاریخ انتشار : 1391/06/01

کلید واژه: Multiplex PCR, شیر گاومیش, استافیلوکوکوس آورئوس, ژن آنتروتوکسین,

چکیده مقاله :

با توجه به اهمیت آنتروتوکسین‌های استافیلوکوکوس موجود در شیر به عنوان یکی از منابع عمده مسمومیت‌های غذایی، بررسی روش‌های متعدد جداسازی، شناسایی و دسته‌بندی این آنتروتوکسین‌ها ضروری است. در این مطالعه توصیفی 75 نمونه باکتری استافیلوکوکوس اورئوس جدا شده از شیر گاومیش برای بررسی وجود و حضور ژن های مربوط به آنتروتوکسین‌های معمول استافیلوکوکوس آرئوس با استفاده از روشMultiplex PCRمورد ارزیابی قرار گرفتند. بدین صورت که ابتدا DNA نمونه‌ها استخراج سپس برای اطمینان از استافیلوکوکوس بودن تمام نمونه‌ها به روش PCR مورد تأیید قرار گرفتند. نتایج حاصل نشان داد که از این تعداد نمونه باکتریایی یک جدایه دارای هر دو باند آنتروتوکسین هایseb و آنتروتوکسینsecبود و سه جدایه دیگر فقط دارای آنتروتوکسین sec بودند و ژن مربوط به آنتروتوکسین A در هیچکدام از جدایه ها شناسایی نشد. بر اساس نتایج این تحقیق میزان شیوع تنوع ژن های مربوط به آنتروتوکسین های معمول در استافیلوکوکوس آرئوس های جدا شده از شیر گاومیش در شهرستان تبریز پایین می باشد.

چکیده انگلیسی:

   Isolation, identification and grouping of staphylococcal enterotoxins in milk is of great importance, since these toxins are considered as a major potential source of humans illness. In this study, 75 staphylococcus aureus strains isolated from buffalo milk were assessed. For this, extracted DNA was analysed by multiplex-PCR for the presence and diversity of genes encoding the common classical staphylococcal enterotoxins. Results revealed that one of the isolates haboured both sec and seb genes; meanwhile, three other isolates contained merely sec gene. Moreover, the gene encoding the production of enterotoxin-A was not detected in any of the isolates. It was concluded that the diversity of genes encoding the classical enterotoxins of S. aureus isolated from Tabriz area was low.

منابع و مأخذ:

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• Abbar, M. (2010). Detection of enterotoxigenic Staphylococcus aureus isolates in domestic dairy products. Iranian Journal of Microbiology, 2(3): 137-142.


• Ahari, H., Shahbazzadeh, D., Misaghi, A. and Pourshafie, M. (2008). Detection and Antibiogram of Staphylococcus aureus in Non Pasteurized Milk Samples from Tehran Province Restricts And Compare to Bacterial Culture. Journal Veterinary Research, 63(5): 321-326.


• Andrea, S., Leonardo, A., Maria, C.F., Cosima, A., Christoph, L. and Roberto, R. (2004). Occurrence of enterotoxin genes and macrorestriction analysis of Staphylococcus aureus isolated from bovine mastitis and bulk-tank milk amples in Italy. An epidemiological study. Italian Journal, 3: 47-53.


• Aragon-Alegro, L.C., Konta, E.M., Suzuki, K., Silva, M.G., Fernandes Junior, A., Raal, R. and Rall, V.L.M. (2007). Occurrence of coagulase positive Staphylococcus in various food products commercialized in Botucatu, SP, Brazil and detection of toxins from food and isolated strains. Food Control, 18: 630-634.


• Baron, E.J., Peterson, L.R. and Finegold, S.M. (1994). Baily and Scotts Diagnostic Microbiology, 9^th Edition, Mosbey-Pear book, Inc, pp. 321-332.


• Bennet, R.W. (1986). Staphylococcus aureus identification characteristics and enterotoxigenicity. Journal of Food science, 51: 1337-39.


• Borelli, B.M., Ferreira, E.G., Lacerda, I.C.A., Santos, D.A., Carmo,L.S., Dias, R.S., Silva, M.C.C. (2006). Enteroxigenic Staphylococcus spp. and other microbial contaminants during production of canastra cheese, Brazil. Brazilian Journal of Microbiology, 37: 545-550.


• da Silva, E.R., do Carmo, L.S. and da Silva, N. (2005). Detection of the enterotoxins A, B, and C genes in Staphylococcus aureus from goat and bovine mastitis in Brazilian dairy herds. Veterinary Microbiology, 106: 103-107.


• Rahimi, E., Mommtaz, H., Shakerian, A., Kavyani, H.R. (2012). The detection of classical enterotoxins of staphylococcus aureus in raw cow milk using the ELISA method. Turk. J. 36(3): 319-322.


• Gilmour, A. and Harvey, J. (1990). Staphylococci in milk and milk products. Journal of Applied Microbiology, 147s-166s.


• Iandolo, J. (1989). Genetic analysis of extracellular toxins of Staphylococcus aureus. Annual Reve Microbiology, 43: 375-402.


• Kapur, V., Sischo, W.M., Greer, R.S., Whittam, I.s. and Musser, J.M. (1995). Molecular population genetic analysis of Staphylococcus aureus recovered from cows. Journal of Clinical Microbiology, 33(2): 376-380.


• Mehrotra, M., Wang, G., Johnson, W.M. (2000). Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1, and methicillin Resistance. Journal of Clinical Microbiology, 38: 1032-1035.


• 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. Veterinary Microbiology,124: 66-72.


• Normanno, G., Firinu, A., Virgilio, S., Mula, G., Dambrosio, A., Poggiu, A., Normanno, G., Firinu, A., Virgilio, S., Mula, G., Dambrosio, A., Poggiu, A., Decastelli, L., Mioni, R., Scuota, S., Bolzoni, G., Di Giannatale, E., Salinetti, A.P., La Salandra, G., Bartoli, M., Zuccon, F., Pirino, T., Sias, S., Parisi, A., Quaglia, N.C. and Celano, G.V. (2005). Coagulase-positive Staphylococci and Staphylococcus aureus in food products marketed in Italy. International Journal of Food Microbiology, 98(1): 73-79.


• Rahimi, E. (2011). Detection of classical enterotoxins of Staphylococcus aureus in raw sheep, goat, camel, and water buffalo milk by ELISA method. Comparative clinical patholology, 1383-4.


• Roberson, J.R., Fox, L.K., Hancock, D.D., Gay, J.M. and Besser, T.E. (1996). Prevalence of coagulase-Posittive staphylococci other than staphylococcus aureus, in bovine mastitis. American Journal of Veterinary Research, 57(1): 54-58.


• Shimi, A. (1996). Veterinary bacteriology and bacterial disease, Jahad publishing. pp, 85-94 [In Farsi].


• Shivram, S. and kelar, S.S. (1985). Compartire Study of markers of pathogenic staphylococci. Indian Journal of Medical Research, 82: 194.


• Soleimani Rahbar, A.K. and Maleknejad, P. (2006). Medical Microbiology. Tehran University Press [In Farsi].


• Quinn, P.J., Markey, B.K., Carter, M.E., Donnelly, W.J. and Leonard, F.C. (2003). Veterinary microbiology and microbial disease. Blackwell publishing.

 

 

• Abbar, M. (2010). Detection of enterotoxigenic Staphylococcus aureus isolates in domestic dairy products. Iranian Journal of Microbiology, 2(3): 137-142.


• Ahari, H., Shahbazzadeh, D., Misaghi, A. and Pourshafie, M. (2008). Detection and Antibiogram of Staphylococcus aureus in Non Pasteurized Milk Samples from Tehran Province Restricts And Compare to Bacterial Culture. Journal Veterinary Research, 63(5): 321-326.


• Andrea, S., Leonardo, A., Maria, C.F., Cosima, A., Christoph, L. and Roberto, R. (2004). Occurrence of enterotoxin genes and macrorestriction analysis of Staphylococcus aureus isolated from bovine mastitis and bulk-tank milk amples in Italy. An epidemiological study. Italian Journal, 3: 47-53.


• Aragon-Alegro, L.C., Konta, E.M., Suzuki, K., Silva, M.G., Fernandes Junior, A., Raal, R. and Rall, V.L.M. (2007). Occurrence of coagulase positive Staphylococcus in various food products commercialized in Botucatu, SP, Brazil and detection of toxins from food and isolated strains. Food Control, 18: 630-634.


• Baron, E.J., Peterson, L.R. and Finegold, S.M. (1994). Baily and Scotts Diagnostic Microbiology, 9^th Edition, Mosbey-Pear book, Inc, pp. 321-332.


• Bennet, R.W. (1986). Staphylococcus aureus identification characteristics and enterotoxigenicity. Journal of Food science, 51: 1337-39.


• Borelli, B.M., Ferreira, E.G., Lacerda, I.C.A., Santos, D.A., Carmo,L.S., Dias, R.S., Silva, M.C.C. (2006). Enteroxigenic Staphylococcus spp. and other microbial contaminants during production of canastra cheese, Brazil. Brazilian Journal of Microbiology, 37: 545-550.


• da Silva, E.R., do Carmo, L.S. and da Silva, N. (2005). Detection of the enterotoxins A, B, and C genes in Staphylococcus aureus from goat and bovine mastitis in Brazilian dairy herds. Veterinary Microbiology, 106: 103-107.


• Rahimi, E., Mommtaz, H., Shakerian, A., Kavyani, H.R. (2012). The detection of classical enterotoxins of staphylococcus aureus in raw cow milk using the ELISA method. Turk. J. 36(3): 319-322.


• Gilmour, A. and Harvey, J. (1990). Staphylococci in milk and milk products. Journal of Applied Microbiology, 147s-166s.


• Iandolo, J. (1989). Genetic analysis of extracellular toxins of Staphylococcus aureus. Annual Reve Microbiology, 43: 375-402.


• Kapur, V., Sischo, W.M., Greer, R.S., Whittam, I.s. and Musser, J.M. (1995). Molecular population genetic analysis of Staphylococcus aureus recovered from cows. Journal of Clinical Microbiology, 33(2): 376-380.


• Mehrotra, M., Wang, G., Johnson, W.M. (2000). Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1, and methicillin Resistance. Journal of Clinical Microbiology, 38: 1032-1035.


• 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. Veterinary Microbiology,124: 66-72.


• Normanno, G., Firinu, A., Virgilio, S., Mula, G., Dambrosio, A., Poggiu, A., Normanno, G., Firinu, A., Virgilio, S., Mula, G., Dambrosio, A., Poggiu, A., Decastelli, L., Mioni, R., Scuota, S., Bolzoni, G., Di Giannatale, E., Salinetti, A.P., La Salandra, G., Bartoli, M., Zuccon, F., Pirino, T., Sias, S., Parisi, A., Quaglia, N.C. and Celano, G.V. (2005). Coagulase-positive Staphylococci and Staphylococcus aureus in food products marketed in Italy. International Journal of Food Microbiology, 98(1): 73-79.


• Rahimi, E. (2011). Detection of classical enterotoxins of Staphylococcus aureus in raw sheep, goat, camel, and water buffalo milk by ELISA method. Comparative clinical patholology, 1383-4.


• Roberson, J.R., Fox, L.K., Hancock, D.D., Gay, J.M. and Besser, T.E. (1996). Prevalence of coagulase-Posittive staphylococci other than staphylococcus aureus, in bovine mastitis. American Journal of Veterinary Research, 57(1): 54-58.


• Shimi, A. (1996). Veterinary bacteriology and bacterial disease, Jahad publishing. pp, 85-94 [In Farsi].


• Shivram, S. and kelar, S.S. (1985). Compartire Study of markers of pathogenic staphylococci. Indian Journal of Medical Research, 82: 194.


• Soleimani Rahbar, A.K. and Maleknejad, P. (2006). Medical Microbiology. Tehran University Press [In Farsi].


• Quinn, P.J., Markey, B.K., Carter, M.E., Donnelly, W.J. and Leonard, F.C. (2003). Veterinary microbiology and microbial disease. Blackwell publishing.