بررسی میزان آلودگى گوشت گاوهاى ذبح‌شده در کشتارگاه صنعتى ارومیه به انگل سارکوسیست با روش PCR و مقایسه با دو روش ماکروسکوپی و هضمی
محورهای موضوعی :
بهداشت مواد غذایی
فاطمه قاسمی کهریزه
1
(دانش آموخته دکترای دامپزشکی، دانشکده دامپزشکی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران)
عباسعلی مطلبی مغانجوقی
2
(استاد گروه بهداشت مواد غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران)
سهراب رسولی
3
(دانشیار گروه انگل‌شناسی، دانشکده دامپزشکی، واحد ارومیه، دانشگاه آزاد اسلامی، ارومیه، ایران)
تاریخ دریافت : 1399/10/11
تاریخ پذیرش : 1400/01/19
تاریخ انتشار : 1399/10/01
کلید واژه:
گاو,
PCR,
سارکوسیست,
ارومیه,
روش ماکروسکوپیک,
روش میکروسکوپیک,
چکیده مقاله :
سارکوسیستیس (Sarcocystis sp.) یکى از مهم‌ترین تک یاخته هاى متعلق به شاخه Apicomplexa مى باشد که در بین حیوانات خونگرم در کلیه نقاط جهان شایع بوده و داراى اهمیت زئونوتیک مى باشد. سه گونه Sarcocystis bovicanis، S. bovifelis و S. hominis در گاو شناسایى شده است. با توجه به بالا بودن میزان آلودگى لاشه هاى گاوهاى کشتارى در ایران به سارکوسیست، میزان آلودگى گوشت گاوهاى ذبح‌شده در کشتارگاه صنعتى ارومیه به این انگل با استفاده از روش مولکولی PCR بررسی شد و کارایی این روش با دو روش ماکروسکوپی و هضمی (میکروسکوپی) در تشخیص میزان آلودگی به این انگل مقایسه شد. در مجموع ٨٠ نمونه از زبان و مری اخذ شده از 40 لاشه مورد مطالعه قرار گرفت. برای بررسی مولکولی، DNA انگل با دستورالعمل کیت استخراج شد و قطعه ژن 18sRNA با استفاده از پرایمر هاى اختصاصى تکثیر شد و در ادامه با آنزیم هاى محدودکننده محصولPCR هضم شد و الگوى شکسته شدن آن‌ها مورد ارزیابى قرار گرفتند. نتایج تحقیق حاضر نشان داد که از مجموع ٤٠ رأس گاو نمونه بردارى شده، در روش هاى ماکروسکوپیک و میکروسکوپیک میزان آلودگى به‌ترتیب 5/2 و 5/72 درصد بود. هم‌چنین تعداد 36 رأس گاو ازلحاظ آلودگى با روش PCR مثبت گزارش گردید که 90 درصد کل گاوها را شامل مى شود. نتایج تحقیق حاضر نشان داد قدرت تشخیص روش مولکولی بیشتر از دو روش دیگر می باشد و میزان آلودگی بافت زبان در هر دو جنسیت نر و ماده بیشتر (05/0< P) از میزان آلودگی بافت مری است. می‌توان به این جمع‌بندی رسید که برای ردیابی موثر سارکوسیست، امکان استفاده از روش PCR به موازات روش‌های متداول وجود دارد.
چکیده انگلیسی:
Sarcocystis is one of the most important protozoa belonging to the phylum Apicomplexa, which is common among warm-blooded animals in all parts of the world, some of which are of zoonotic importance. Sarcocystis cruzi, S. bovifelis, and S. hominis are recognized in cattle. Due to the high occurrence of Sarcocystis in cow carcasses slaughtered in Iran, this study was conducted to investigate the contamination of Sarcocystis in beef slaughtered in Urmia industrial slaughterhouse using PCR. Also, the efficiency of the PCR method was compared with macroscopic and digestive (microscopic) methods. For this, a total of 80 esophageal and tongue samples, obtained from 40 carcasses was assayed. The Sarcocystis DNA was extracted according to the instructions of the Qiagen kit and the 18sRNA gene fragment was used using specific primers. The DNA product was digested with restriction enzymes, and their fracture pattern was evaluated. Out of 40 carcasses, in macroscopic and microscopic methods 2.5% and 72.5% were found positive for Sarcocystis, respectively. Besides, 36 cows were reported positive in terms of PCR contamination, which is 90% of the total samples. The results showed that the efficiency of PCR in detecting Sarcocystis is higher than the other two methods (p < /em><0.05). Moreover, it was revealed that the tongue contamination in both male and female cattle carcass was higher (p < /em>< 0.05) than the level of esophageal contamination. It was concluded that for efficient detection of Sarcocystis in cattle carcass, PCR can be applied along with the conventional methods.
منابع و مأخذ:
· Alibeigi, Z., Rahbari, S., Hoghooghirad, N. and Naisi, S. (2015). Macroscopic and microscopic survey of Sarcocystosis in ruminants Shahriar slaughterhouse during 2012-2013. Journal of Veterinary Research, 70(4): 441-445. [In Persian]
· Ali Hemmati, A., Shahbazi, A., Fallah, E. and Esfaram, Sh. (2013). Prevalence of Sarcocystis infection in slaughtered cows in Tabriz. Journal of Comparative Pathobiology, 10(4): 1095-1100. [In Persian]
· Arshad, M., Dalimi, A. and Ghaffarifar F. (2007). Comparative study on Sarcocystis diagnosis in meat of slaughtered sheep in Tabriz. Pajouhesh and Sazandegi, 75: 68-72. [In Persian]
· Berenji, F., Behniafar, H., Zabolinejad, N., Fata, A.M., Salehi, M. and Sadabadi, F. (2019). Prevalence of Sarcocystis infection in slaughtered sheep by macroscopic and histopathologic method in Mashhad. Medical Journal of Mashhad University of Medical Sciences, 62(3): 1556-1561. [In Persian]
· Beyazit, A., Yazicioglu, O. and Karear, Z. (2007). The prevalence of ovine Sarcocystis species in Izmir province. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 54: 111-116.
· Bonyadin, M. and Mashki, B. (2006). Evaluation of contamination of carcasses of cows slaughtered in Shahrekord slaughterhouse to sarcophagus. Research and Construction, 14-18. [In Persian]
· Dalimi, A., Paykari H., Esmaeilzadeh, M., Valizadeh, M., Karimi, G.H. , Motamedi, G.H. et al. (2008). Identification of Sarcocystis species of slaughtered sheep in Gazvin Ziaran slaughterhouse using PCR-RFLP. Pathobiology Research (Modares Journal of Medical Science), 11(1 &2): 65-72. [In Persian]
· Domenis, L., Peletto, S., Sacchi, L., Clementi, E., Genchi, M., Felisari, L. et al. (2011). Detection of a morphogenetically novel Sarcocystis hominis- like in the context of a prevalence study in semi-intensively bred cattle in Italy. Parasitology Research, 109(6): 1677-1687.
· Dubey, J.P., Speer, C.A. and Fayer, R. (1989a). Sarcocystis of animal and man. CRC Press, Inc., Florida, pp. 62-78.
· Dubey, J.P., Speer, C.A. and Charleston, W.A.G. (1989b). Ultrastructural differentiation between Sarcocystis of Sarcocystis hirsuta and S. hominis. Veterinary Parasitology, (34): 153–157.
· Eslami, G., Zohourtabar, A. and Mehrizi, S.R. (2014). First molecular identification of Sarcocystis hirsuta in Iranian beef: A case report. Journal of Food Quality and Hazards Control, 1(1): 32-34.
· Fayer, R. (2004). Sarcocystis spp. in human infections. Clinical Microbiological Review, 17 (4): 894-902.
· Fayer, R., Esposito, D.H. and Dubey, J.P. (2015). Human infections with Sarcocystis species. Clinical Microbiology Reviews. 28: 295–311.
· Hamidinejat, H., Moetamedi, H., Alborzi, A. and Hatami, A. (2014). Molecular detection of Sarcocystis species in slaughtered sheep by PCR–RFLP from south-western of Iran. Journal of Parasitic Diseases, 38(2): 233–237.
· Imre, K., Morariu, S., Imre, M., Plutzer, J., Boldea, M. and Morar, A. (2019). High prevalence of Sarcocystis spp. infections in cattle (Bos taurus) from central China. Parasitology research, 67: 800-804.
· Kalantari, N., Bayani, M. and Ghaffari, S. (2013). Sarcocystis cruzi: first molecular identification from cattle in Iran. International Journal of Molecular and Cellular Medicine, 2(3): 130-126.
· Kargar Jahromi, Z., Solhjoo, K., Zareian Jahromi, M., Kargar Jahromi, H., Erfaniyan, S. et al., (2012). Investigation of Sarcocystis infection in slaughtered goats in Jahrom abattoir. Journal of Fasa University of Medical Sciences, 2(3): 163-167. [In Persian]
· Mandaalee, A. and Rasouli, S. (2020). Survey on the contamination rate of Sarcocystis in slaughtered sheep by digestive method. Journal of Food Hygiene, 10(3): 31-41. [In Persian]
· Mirzaei Dehaghi, M., Fallahi, M., Sami, M. and Radfar, M.H. (2012) Survey of Sarcocystis infection in slaughtered sheep in Kerman abattoir, Kerman, Iran. Comparative Clinical Pathology, 22: 343-346.
· Nematollahia, A., Khoshkerdar, A., Ashrafi Helan, J., Shahbazi, P. and Hassanzadeh, P. (2013). A study on rate of infestation to Sarcocystis cysts in supplied raw hamburgers. Journal of Parasitic Diseases, 39(2): 276–279.
· Obijiaku, I.N., Ajogi, I., Umoh, J.U., Lawal, I.A. and Atu, B.O. (2013). Sarcocystis infection in slaughtered cattle in Zango abattoir, Zaria, Nigeria. Veterinary World, 6(6): 346-349.
· Oryan, A., Ahmadi, N. and Mousavi, S.M. (2010). Prevalence, biology, and distribution pattern of Sarcocystis infection in water buffalo (Bubalus bubalis) in Iran. Tropical Animal Health and Production, 42(7): 1513-1518.
· Parandian, F., Feizi, F., Maghsood, A.H., Matini, M., Roshan, A. and Fallah, M. (2015). A Survey on Sarcocystis infection rate in slaughtered cattle and sheep by macroscopic inspection and pepsin digestion methods in Hamadan abattoir. Scientific Journal of Hamadan University of Medical Sciences, 22(3): 210-216.
· Rahimi, E., Dousti, A., Hosseini, S.R., Homayoun, M. and Abdizadeh, R. (2012). Identify of Sarcocystis hominis in cattle carcasses using PCR method. Comparative Pathobiology, 9(2): 661-664. [In Persian]
· Rasouli, S., Ahari, H., Farsi, A. and Amin Khah, S. (2006). Evaluation of meat contamination in butchers of Urmia city to Sarcocyst protozoan by digestion method. Journal of Veterinary Microbiology, 5(4): 1-5. [In Persian]
· Rasouli, S., Sedghian, M., Karimian, S., Valizadeh, E. and Jafari, K. (2010). Prevalence of protozoa Sarcocystis contamination of meat in the city of Sanandaj with digestion method. Journal of New Veterinary Research, 1(3): 27-32. [In Persian]
· Razmi, G. and Rahbari, S. (2000). Study of Sarcocystis in the domestic ruminants of Tehran and Golestan provine. Journal of Veterinary Faculty, Shahid Chamran University, 40: 39-46. [In Persian]
· Sayed, F.G., Shaheen, M.S.I., Arafa, M.I. and Koraa, H.M. (2008). Sarcocystis infection in cattle at Assiute abattoir, microscopical and serological studies. Bullding and Environment Research, 11(1): 47-58.
· Shekar Foroush, S.Sh. and Ahmadi, B. (2004). Contamination of Sarcocystis carcasses of slaughtered cows in Isfahan slaughterhouse and its health importance. Pajouhesh and Sazandegi, 64: 102-103. [In Persian]
· Tenter, A.M. (1995). Current research on Sarcocystis species of domestic animals. International Journal for Parasitology, 25: 1311-1330.
· Vangeel, L., Houf, K., Chiers, K., Vercruysse, J., Herde, K. and Ducatelle, R. (2007). Molecular-based identification of Sarcocystis hominis in Belgian minced beef. Journal of Food Protection, 67: 800-804.
· Yakhchali, M., Morshedi, A. and Malekifard, F. (2010). Screening of seropositive Sarcocystis (Apicomplexa: Sarcocystidae, Lankester1882) in sheep using counterim-munoelectrophoresis. Veterinary Journal (Pajouhesh and Sazandegi), 89: 28-32. [In Persian]
· Yang, Y., Dong, H., Su, R., Wang, Y., Wang, R. and Jiang, Y. (2018). High prevalence of Sarcocystis spp. infections in cattle (Bos taurus) from central China. Parasitology research, 67 (6): 800-804.
· Zuo, Y. (1992). Coccidia and coccidiosis of domestic animals and man. Tian Jing China: Tian Jing Scientific and Technical Publishing House, p. 125.
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· Alibeigi, Z., Rahbari, S., Hoghooghirad, N. and Naisi, S. (2015). Macroscopic and microscopic survey of Sarcocystosis in ruminants Shahriar slaughterhouse during 2012-2013. Journal of Veterinary Research, 70(4): 441-445. [In Persian]
· Ali Hemmati, A., Shahbazi, A., Fallah, E. and Esfaram, Sh. (2013). Prevalence of Sarcocystis infection in slaughtered cows in Tabriz. Journal of Comparative Pathobiology, 10(4): 1095-1100. [In Persian]
· Arshad, M., Dalimi, A. and Ghaffarifar F. (2007). Comparative study on Sarcocystis diagnosis in meat of slaughtered sheep in Tabriz. Pajouhesh and Sazandegi, 75: 68-72. [In Persian]
· Berenji, F., Behniafar, H., Zabolinejad, N., Fata, A.M., Salehi, M. and Sadabadi, F. (2019). Prevalence of Sarcocystis infection in slaughtered sheep by macroscopic and histopathologic method in Mashhad. Medical Journal of Mashhad University of Medical Sciences, 62(3): 1556-1561. [In Persian]
· Beyazit, A., Yazicioglu, O. and Karear, Z. (2007). The prevalence of ovine Sarcocystis species in Izmir province. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 54: 111-116.
· Bonyadin, M. and Mashki, B. (2006). Evaluation of contamination of carcasses of cows slaughtered in Shahrekord slaughterhouse to sarcophagus. Research and Construction, 14-18. [In Persian]
· Dalimi, A., Paykari H., Esmaeilzadeh, M., Valizadeh, M., Karimi, G.H. , Motamedi, G.H. et al. (2008). Identification of Sarcocystis species of slaughtered sheep in Gazvin Ziaran slaughterhouse using PCR-RFLP. Pathobiology Research (Modares Journal of Medical Science), 11(1 &2): 65-72. [In Persian]
· Domenis, L., Peletto, S., Sacchi, L., Clementi, E., Genchi, M., Felisari, L. et al. (2011). Detection of a morphogenetically novel Sarcocystis hominis- like in the context of a prevalence study in semi-intensively bred cattle in Italy. Parasitology Research, 109(6): 1677-1687.
· Dubey, J.P., Speer, C.A. and Fayer, R. (1989a). Sarcocystis of animal and man. CRC Press, Inc., Florida, pp. 62-78.
· Dubey, J.P., Speer, C.A. and Charleston, W.A.G. (1989b). Ultrastructural differentiation between Sarcocystis of Sarcocystis hirsuta and S. hominis. Veterinary Parasitology, (34): 153–157.
· Eslami, G., Zohourtabar, A. and Mehrizi, S.R. (2014). First molecular identification of Sarcocystis hirsuta in Iranian beef: A case report. Journal of Food Quality and Hazards Control, 1(1): 32-34.
· Fayer, R. (2004). Sarcocystis spp. in human infections. Clinical Microbiological Review, 17 (4): 894-902.
· Fayer, R., Esposito, D.H. and Dubey, J.P. (2015). Human infections with Sarcocystis species. Clinical Microbiology Reviews. 28: 295–311.
· Hamidinejat, H., Moetamedi, H., Alborzi, A. and Hatami, A. (2014). Molecular detection of Sarcocystis species in slaughtered sheep by PCR–RFLP from south-western of Iran. Journal of Parasitic Diseases, 38(2): 233–237.
· Imre, K., Morariu, S., Imre, M., Plutzer, J., Boldea, M. and Morar, A. (2019). High prevalence of Sarcocystis spp. infections in cattle (Bos taurus) from central China. Parasitology research, 67: 800-804.
· Kalantari, N., Bayani, M. and Ghaffari, S. (2013). Sarcocystis cruzi: first molecular identification from cattle in Iran. International Journal of Molecular and Cellular Medicine, 2(3): 130-126.
· Kargar Jahromi, Z., Solhjoo, K., Zareian Jahromi, M., Kargar Jahromi, H., Erfaniyan, S. et al., (2012). Investigation of Sarcocystis infection in slaughtered goats in Jahrom abattoir. Journal of Fasa University of Medical Sciences, 2(3): 163-167. [In Persian]
· Mandaalee, A. and Rasouli, S. (2020). Survey on the contamination rate of Sarcocystis in slaughtered sheep by digestive method. Journal of Food Hygiene, 10(3): 31-41. [In Persian]
· Mirzaei Dehaghi, M., Fallahi, M., Sami, M. and Radfar, M.H. (2012) Survey of Sarcocystis infection in slaughtered sheep in Kerman abattoir, Kerman, Iran. Comparative Clinical Pathology, 22: 343-346.
· Nematollahia, A., Khoshkerdar, A., Ashrafi Helan, J., Shahbazi, P. and Hassanzadeh, P. (2013). A study on rate of infestation to Sarcocystis cysts in supplied raw hamburgers. Journal of Parasitic Diseases, 39(2): 276–279.
· Obijiaku, I.N., Ajogi, I., Umoh, J.U., Lawal, I.A. and Atu, B.O. (2013). Sarcocystis infection in slaughtered cattle in Zango abattoir, Zaria, Nigeria. Veterinary World, 6(6): 346-349.
· Oryan, A., Ahmadi, N. and Mousavi, S.M. (2010). Prevalence, biology, and distribution pattern of Sarcocystis infection in water buffalo (Bubalus bubalis) in Iran. Tropical Animal Health and Production, 42(7): 1513-1518.
· Parandian, F., Feizi, F., Maghsood, A.H., Matini, M., Roshan, A. and Fallah, M. (2015). A Survey on Sarcocystis infection rate in slaughtered cattle and sheep by macroscopic inspection and pepsin digestion methods in Hamadan abattoir. Scientific Journal of Hamadan University of Medical Sciences, 22(3): 210-216.
· Rahimi, E., Dousti, A., Hosseini, S.R., Homayoun, M. and Abdizadeh, R. (2012). Identify of Sarcocystis hominis in cattle carcasses using PCR method. Comparative Pathobiology, 9(2): 661-664. [In Persian]
· Rasouli, S., Ahari, H., Farsi, A. and Amin Khah, S. (2006). Evaluation of meat contamination in butchers of Urmia city to Sarcocyst protozoan by digestion method. Journal of Veterinary Microbiology, 5(4): 1-5. [In Persian]
· Rasouli, S., Sedghian, M., Karimian, S., Valizadeh, E. and Jafari, K. (2010). Prevalence of protozoa Sarcocystis contamination of meat in the city of Sanandaj with digestion method. Journal of New Veterinary Research, 1(3): 27-32. [In Persian]
· Razmi, G. and Rahbari, S. (2000). Study of Sarcocystis in the domestic ruminants of Tehran and Golestan provine. Journal of Veterinary Faculty, Shahid Chamran University, 40: 39-46. [In Persian]
· Sayed, F.G., Shaheen, M.S.I., Arafa, M.I. and Koraa, H.M. (2008). Sarcocystis infection in cattle at Assiute abattoir, microscopical and serological studies. Bullding and Environment Research, 11(1): 47-58.
· Shekar Foroush, S.Sh. and Ahmadi, B. (2004). Contamination of Sarcocystis carcasses of slaughtered cows in Isfahan slaughterhouse and its health importance. Pajouhesh and Sazandegi, 64: 102-103. [In Persian]
· Tenter, A.M. (1995). Current research on Sarcocystis species of domestic animals. International Journal for Parasitology, 25: 1311-1330.
· Vangeel, L., Houf, K., Chiers, K., Vercruysse, J., Herde, K. and Ducatelle, R. (2007). Molecular-based identification of Sarcocystis hominis in Belgian minced beef. Journal of Food Protection, 67: 800-804.
· Yakhchali, M., Morshedi, A. and Malekifard, F. (2010). Screening of seropositive Sarcocystis (Apicomplexa: Sarcocystidae, Lankester1882) in sheep using counterim-munoelectrophoresis. Veterinary Journal (Pajouhesh and Sazandegi), 89: 28-32. [In Persian]
· Yang, Y., Dong, H., Su, R., Wang, Y., Wang, R. and Jiang, Y. (2018). High prevalence of Sarcocystis spp. infections in cattle (Bos taurus) from central China. Parasitology research, 67 (6): 800-804.
· Zuo, Y. (1992). Coccidia and coccidiosis of domestic animals and man. Tian Jing China: Tian Jing Scientific and Technical Publishing House, p. 125.
