جداسازی و شمارش باسیلوس سرئوس از شیرخام عرضه شده در شهر تبریز و جستجوی ژن ces در جدایه‌ها
محورهای موضوعی :
علوم و صنایع غذایی
مهدیه حیدرزاده
1
(دانش آموخته کارشناسی ارشد، گروه میکروبیولوژی، موسسه آموزش عالی ربع رشید، تبریز، ایران)
افشین جوادی
2
(دانشیارگروه بهداشت مواد غذایی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران.)
تاریخ دریافت : 1396/06/16
تاریخ پذیرش : 1396/08/30
تاریخ انتشار : 1397/04/01
کلید واژه:
شیر خام,
باسیلوس سرئوس,
ژن ces,
چکیده مقاله :
باسیلوس سرئوس یک باکتری گرم مثبت، اسپوردار است که انتشار وسیعی در طبیعت دارد و به عنوان یکی از عوامل عمده پاتوژن موادغذایی شناسایی شده است، اغلب در آلودگی غذاهای آماده و فراورده‌های شیر نقش ایفا می‌کند و باعث ایجاد دونوع مسمومیت غذایی: نوع اسهالی و نوع استفراغی می‌شود. هدف از این مطالعه، جداسازی و شمارش باسیلوس سرئوس در شیر خام عرضه شده در فروشگاه‌ها‌ی سطح شهر تبریز و جستجوی ژن ces در جدایه‌ها می‌باشد. برای این منظور، تعداد 120 نمونه شیر خام، طی اسفند ماه 1395 تا خرداد 1396 از فروشگاه‌ها‌ی شهر تبریز به صورت تصادفی ساده انتخاب و تحت شرایط استریل به آزمایشگاه بهداشت موادغذایی منتقل شد تا مورد جستجو، شناسایی و شمارش باسیلوس سرئوس به روش استاندارد ملی ایران و PCR قرار گیرند. طبق نتایج بدست آمده از روش کشت، از تعداد 120 نمونه شیرخام، (83/10 درصد) 13 نمونه آلوده به باکتری باسیلوس سرئوس بودند که میانگین بار آلودگی آن‌ها 8/2 بود. نتایجPCR نشان داد که از 13 نمونه مثبت باسیلوس سرئوس 12 نمونه (3/92 درصد) دارای ژن استفراغ‌زای cesبودند. یافته‌های مطالعه حاضر نشان داد که شیرخام پتانسیل مسمومیت‌زایی به باسیلوس سرئوس را دارد و به کار گرفتن روش‌هایی جهت کاهش آلودگی به این باکتری در جریان فراوری فراورده‌های شیر الزامی می‌باشد.
چکیده انگلیسی:
Bacillus cereus is a gram-positive and spore-forming bacterium which is widely distributed in nature. It also has been known as a major food borne pathogen that often plays a role in the contamination of ready-to-eat and dairy products. It causes two different types of food poisoning in human: the diarrheal type and the emetic type. The aim of this study was to isolate and enumerate B. cereus in raw milk distributed in the stores of Tabriz city, Iran and to detect ces gene among the isolates. For this purpose, 120 samples of raw milk were randomly selected from the stores during February to June 2017. They were transferred to the Laboratory of food hygiene under sterile conditions in order to isolate, identify and enumerate B. cereus according to the national standard method of Iran and PCR. According to the results of microbial culture, out of 120 samples of raw milk, 13 (10.83%) samples were contaminated with B. cereus and the mean rate of contamination was 2.8 log10/ml. PCR results showed that of 13 positive samples, 12 (92.3%) samples contained ces gene which is responsible for vomiting. The results of this study showed that raw milk has the potential for human intoxication by B. cereus; and the use of procedures to reduce the bacterial contamination during the processing of dairy product is required.
منابع و مأخذ:
· Ankolekar, C., Rahmati, T. and Labbe, R.G. (2009). Detection of toxigenic Bacillus cereus and Bacillus thuringiensis spores in US rice. International Journal of Food Microbiology, 128(3), 460-466.
· Cui, Y., Liu, X., Dietrich, R., Martlbauer, E., Cao, J., Ding, S. and et al. (2016). Characterization of Bacillus cereus isolates from local dairy farms in China. Federation of European Microbiological Societies, 363(12).
· Ehling-Schulz, M., Vukov, N., Schulz, A., Shaheen, R., Andersson, M., Märtlbauer, E. and et al. (2005). Identification and partial characterization of the nonribosomal peptide synthetase gene responsible for cereulide production in emetic Bacillus cereus. Applied and Environmental Microbiology, 71(1), 105-113.
· Evelyn, E. and Silva, F.V.M. (2015). Thermosonication versus thermal processing of skim milk and beef slurry modeling the inactivation kinetics of psychrotrophic Bacillus cereus spores. Food Research International, 67: 67–74.
· Gitahi, N.J., Ombui, J.N., Nduati, D.W., Gicheru, M.M. (2009). Genetis characterisation of food borne Bacillus cereus strains from milk, cheese and rice by multiplex pcr assay. Journal of Integrative Biology, 5(2), 82-6.
· Institute of Standards and Industrial Research of Iran. (2006). Counting and identification of Bacillus cereus in food. 2nd revision, ISIRI No. 2324. [In Persian]
· Iran Food and Drug Administration. Customer Microbiology-table limits. (2014). M.5:SOP. No review: 2. [In Persian].
· Jung-Beom, K., Kim, J.M., Park, Y.B., Han, J.A., Lee, S.H. and Kwak, H.S. (2010). Evaluation of Various PCR Assays for Detection of Emetic-Toxin-Producing Bacillus cereus. Journal of Microbiol Biotechnol, 20(7): 1107–1113.
· Kumari, S. and Sarkar, P.K. (2014). In vitro model study for biofilm formation by Bacillus cereus in dairy chilling tanks and optimization of clean-in-place (CIP) regimes using response surface methodology. Journal of Food Control, 36: 153-158.
· Lay, J.L., Bahloul, H., Serino, S., Jobin, M. and Schmitt, PH. (2015). Reducing activity, glucose metabolism and acid tolerance response of Bacillus cereus grown at various pH and oxydo-reduction potential levels. Food Microbiology, 46: 314-321.
· Lues, J., Venter, P. and Van Der Westhuizen, H. (2003). Enumeration of potential microbiological hazards in milk from a marginal urban settlement in central South Africa. Journal of Food Microbiology, 20(3): 321-26.
· Magnusson, M., Christiansson, A. and Svensson, B. (2007). Bacillus cereus Spores during housing of dairy cows factors affecting contamination of raw milk. Journal of Dairy Science, 90: 2745–2754.
· Messaoudi, K., Clavel, T., Schmitt, Ph. and Duport, C. (2010). mediates carbohydrate-dependent regulation of catabolic and enterotoxin genes in Bacillus cereus F4430/73. Research in Microbiology, 61(1): 30-39.
· Messelhausser, U., Kämpf, P., Fricker, M., Ehling-Schulz, M., Zucker, R., Wagner, B. and et al. (2010). Prevalence of emetic Bacillus cereus in different ice creams in Bavaria. Journal of Food Protection, 73(2): 395-399.
· Meybodi, M. and Behzadian Nejad, G. (2009). Isolation of B. cereus from dairy products and detoxification of 20 selected strains. Hakim Research Journal, 2(2): 114-119. [In Persian]
· Molayi Kohneh Shahri, Sh., Hoseinzadegan, H. and Taghi Nejad, J. (2015). A review of the structure of virulence of B.cereus. Quarterly Lab and Recognition, 29: 51-67. [In Persian]
· Moradi-Khatoonabadi, Z., Maghsoudlou, Y., Ezzatpanah, H., Khomeiri, M. and Aminafshar, M. (2014). Occurrence of Bacillus cereus in raw milk receiving from UF-feta cheese plants. Iranian Journal of Health and Environment, 6(4): 545-556.
· Reyes, A.L., Montanhini, M.T., Bittencourt, J.V., Destro, M.T. and Bersot, L.S. (2013). Gene detection and toxin production evaluation of hemolysin BL of Bacillus cereus isolated from milk and dairy products marketed in Brazil. Brazilian Journal of Microbiology, 44(4): 1195-1198.
· Samapundo, S., Heyndrickx, M., Xhaferi, R. and Devlieghere, F. (2011). Incidence, diversity and toxin gene characteristics of Bacillus cereus group strains isolated from food products marketed in Belgium. International Journal of Food Microbiology, 150(1): 34-41.
· Schoeni, J.L. and Wong, A.C.L. (2005). Bacillus cereus food poisoning and its toxins. Journal of Food Prot, 68(3): 636-648.
· Shaheen, R., Svensson, B., Andersson, M.A., Christiansson, A. and Salkinoja-Salonen, M. (2010). Persistence strategies of Bacillus cereus spores isolated from dairy silo tanks. Food Microbiology, 27: 347-355.
· Soleimaninanadegani, M. (2013). Evaluation of critical level in Bacillus Cereus growth curve in milk products with different conditions based on experimental data and combase predictive models, Journal of Webmed Central , 4(9): 2046-1690.
· Verraes, C., Vlaemynck, G., Weyenberg, S.V., Zutter, L.D., Daube, G. and Sindic, M. (2015). A review of the microbiological hazards of dairy products made from raw milk. International Dairy Journal, 50: 32-44.
· Williams, A.G. and Withers, S.E. (2010). Microbiological 9haracterization of artisanal farmhouse cheeses manufactured in Scotland. International Journal of Dairy Technology, 63: 356-369.
· Zhang, Z., Wang, L., Xu, H., Aguilar, Z. P., Liu, C., Gan, B. and et al. (2014). Detection of non-emetic and emetic Bacillus cereus by propidium monoazide multiplex PCR (PMA-mPCR) with internal amplification control. Food Control, 35(1): 401-406.
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· Ankolekar, C., Rahmati, T. and Labbe, R.G. (2009). Detection of toxigenic Bacillus cereus and Bacillus thuringiensis spores in US rice. International Journal of Food Microbiology, 128(3), 460-466.
· Cui, Y., Liu, X., Dietrich, R., Martlbauer, E., Cao, J., Ding, S. and et al. (2016). Characterization of Bacillus cereus isolates from local dairy farms in China. Federation of European Microbiological Societies, 363(12).
· Ehling-Schulz, M., Vukov, N., Schulz, A., Shaheen, R., Andersson, M., Märtlbauer, E. and et al. (2005). Identification and partial characterization of the nonribosomal peptide synthetase gene responsible for cereulide production in emetic Bacillus cereus. Applied and Environmental Microbiology, 71(1), 105-113.
· Evelyn, E. and Silva, F.V.M. (2015). Thermosonication versus thermal processing of skim milk and beef slurry modeling the inactivation kinetics of psychrotrophic Bacillus cereus spores. Food Research International, 67: 67–74.
· Gitahi, N.J., Ombui, J.N., Nduati, D.W., Gicheru, M.M. (2009). Genetis characterisation of food borne Bacillus cereus strains from milk, cheese and rice by multiplex pcr assay. Journal of Integrative Biology, 5(2), 82-6.
· Institute of Standards and Industrial Research of Iran. (2006). Counting and identification of Bacillus cereus in food. 2nd revision, ISIRI No. 2324. [In Persian]
· Iran Food and Drug Administration. Customer Microbiology-table limits. (2014). M.5:SOP. No review: 2. [In Persian].
· Jung-Beom, K., Kim, J.M., Park, Y.B., Han, J.A., Lee, S.H. and Kwak, H.S. (2010). Evaluation of Various PCR Assays for Detection of Emetic-Toxin-Producing Bacillus cereus. Journal of Microbiol Biotechnol, 20(7): 1107–1113.
· Kumari, S. and Sarkar, P.K. (2014). In vitro model study for biofilm formation by Bacillus cereus in dairy chilling tanks and optimization of clean-in-place (CIP) regimes using response surface methodology. Journal of Food Control, 36: 153-158.
· Lay, J.L., Bahloul, H., Serino, S., Jobin, M. and Schmitt, PH. (2015). Reducing activity, glucose metabolism and acid tolerance response of Bacillus cereus grown at various pH and oxydo-reduction potential levels. Food Microbiology, 46: 314-321.
· Lues, J., Venter, P. and Van Der Westhuizen, H. (2003). Enumeration of potential microbiological hazards in milk from a marginal urban settlement in central South Africa. Journal of Food Microbiology, 20(3): 321-26.
· Magnusson, M., Christiansson, A. and Svensson, B. (2007). Bacillus cereus Spores during housing of dairy cows factors affecting contamination of raw milk. Journal of Dairy Science, 90: 2745–2754.
· Messaoudi, K., Clavel, T., Schmitt, Ph. and Duport, C. (2010). mediates carbohydrate-dependent regulation of catabolic and enterotoxin genes in Bacillus cereus F4430/73. Research in Microbiology, 61(1): 30-39.
· Messelhausser, U., Kämpf, P., Fricker, M., Ehling-Schulz, M., Zucker, R., Wagner, B. and et al. (2010). Prevalence of emetic Bacillus cereus in different ice creams in Bavaria. Journal of Food Protection, 73(2): 395-399.
· Meybodi, M. and Behzadian Nejad, G. (2009). Isolation of B. cereus from dairy products and detoxification of 20 selected strains. Hakim Research Journal, 2(2): 114-119. [In Persian]
· Molayi Kohneh Shahri, Sh., Hoseinzadegan, H. and Taghi Nejad, J. (2015). A review of the structure of virulence of B.cereus. Quarterly Lab and Recognition, 29: 51-67. [In Persian]
· Moradi-Khatoonabadi, Z., Maghsoudlou, Y., Ezzatpanah, H., Khomeiri, M. and Aminafshar, M. (2014). Occurrence of Bacillus cereus in raw milk receiving from UF-feta cheese plants. Iranian Journal of Health and Environment, 6(4): 545-556.
· Reyes, A.L., Montanhini, M.T., Bittencourt, J.V., Destro, M.T. and Bersot, L.S. (2013). Gene detection and toxin production evaluation of hemolysin BL of Bacillus cereus isolated from milk and dairy products marketed in Brazil. Brazilian Journal of Microbiology, 44(4): 1195-1198.
· Samapundo, S., Heyndrickx, M., Xhaferi, R. and Devlieghere, F. (2011). Incidence, diversity and toxin gene characteristics of Bacillus cereus group strains isolated from food products marketed in Belgium. International Journal of Food Microbiology, 150(1): 34-41.
· Schoeni, J.L. and Wong, A.C.L. (2005). Bacillus cereus food poisoning and its toxins. Journal of Food Prot, 68(3): 636-648.
· Shaheen, R., Svensson, B., Andersson, M.A., Christiansson, A. and Salkinoja-Salonen, M. (2010). Persistence strategies of Bacillus cereus spores isolated from dairy silo tanks. Food Microbiology, 27: 347-355.
· Soleimaninanadegani, M. (2013). Evaluation of critical level in Bacillus Cereus growth curve in milk products with different conditions based on experimental data and combase predictive models, Journal of Webmed Central , 4(9): 2046-1690.
· Verraes, C., Vlaemynck, G., Weyenberg, S.V., Zutter, L.D., Daube, G. and Sindic, M. (2015). A review of the microbiological hazards of dairy products made from raw milk. International Dairy Journal, 50: 32-44.
· Williams, A.G. and Withers, S.E. (2010). Microbiological 9haracterization of artisanal farmhouse cheeses manufactured in Scotland. International Journal of Dairy Technology, 63: 356-369.
· Zhang, Z., Wang, L., Xu, H., Aguilar, Z. P., Liu, C., Gan, B. and et al. (2014). Detection of non-emetic and emetic Bacillus cereus by propidium monoazide multiplex PCR (PMA-mPCR) with internal amplification control. Food Control, 35(1): 401-406.
