شناسایی میزان آلودگی به سارکوسیستیس به روش مولکولی و با استفاده از ژن هایCOX1 و 18srRNA در گاو و گاومیش های شهرستان ارومیه

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه ارومیه، ارومیه، ایران.

2 گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه ارومیه، ارومیه، ایران

چکیده

با توجه به اهمیت سارکوسیتوزیس به عنوان بیماری مشترک در بهداشت عمومی جامعه، در این مطالعه میزان فراونی گونه‌های سارکوسیستیس با استفاده از ژن‌های Cox1 و 18srRNA در گاوها و گاومیش‌های کشتارشده در گشتارگاه ارومیه مورد بررسی قرار گرفت. برای این منظور، نمونه‌برداری از عضلات بین دنده‌ای، دیافراگم، مری و ران 100 راس گاو و 100 راس گاومیش کشتار شده در کشتارگاه شهرستان ارومیه در سال 1400 صورت گرفت. نمونه‌ها از نظر ماکروسکوپی و میکروسکوپی با استفاده از روش گسترش مهری و هضم بافتی مورد بررسی قرار گرفت. در ادامه استخراج DNA  از بافت های مورد مطالعه صورت گرفت و سپس بررسی‌های مولکولی با استفاده از ژن‌های COX1 و 18srRNA انجام گرفت. در بررسی ماکروسکوپی، هیچکدام از نمونه‌های بدست آمده از گاو آلوده به سارکوسیستیس نبودند، در حالی که تک یاخته از 11 (11درصد) نمونه بدست آمده از گاومیش جداسازی شد. همچنین در بررسی‌های میکروسکوپی با استفاده از روش گسترش مهری میزان آلودگی در گاو وگاومیش به ترتیب 27 (27 درصد) و 16 (16درصد) و در روش هضم بافتی به ترتیب برابر با 37 (37 درصد) و 23 (23 درصد) بود. همچنین بررسی‌های مولکولی با استفاده از ژن‌های COX1 و 18srRNA، حاکی از تایید آلودگی به سارکوسیتیس در 39مورد (39درصد) از 100 لاشه گاو و 26 مورد (26درصد) از 100 لاشه گاومیش مورد مطالعه بود. یافته‌های این مطالعه، بیانگر آلودگی نسبتاً بالا در جمعیت‌های گاو وگاومیش استان دارد. به همین دلیل اقدامات و توصیه‌های لازم باید در جهت جلوگیری از مصرف گوشت خام و کم پخته در نظر گرفته شود.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Detection of sarcocystis infection by molecular methods using COX 1 and 18srRNA genes in cattle and buffaloes of Urmia

نویسندگان [English]

  • Shima Abdolrahmani, 1
  • Farnaz Malekifard 2
  • Bijan Esmaeilnejad 1
  • Mousa Tavassoli 1
1 Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
2 Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
چکیده [English]

Due to the importance of Sarcocystis as a common zoonotic disease in public health, in this study, the infection rate of Sarcocystis infection and molecular identification of its species by using COX1 and 18srRNA genes in slaughtered cattle and buffaloes of the abattoir of Urmia was investigated. For this purpose, samples were taken from the intercostal muscles, diaphragm, esophagus, and thighs of 100 cows and 100 buffaloes slaughtered in the slaughterhouse of Urmia city in 2021. At first, macroscopic and microscopic evaluation were performed on samples. Then, DNA was extracted from homogenized tissues and molecular investigations were done using COX1 and 18srRNA genes. In the macroscopic examination, none of the samples obtained from cows were infected with sarcocystis, while this protozoan was isolated from 11 (%11) samples obtained from buffaloes. The infection rate in the impression smear method in cow and buffaloes samples were 27 (%27) and 16 (%16) and in peptic digestion method was 37 (%37) and 23 (%23), respectively. In parallel, molecular evaluation confirmed the presence of sarcocystis infection in 39 cases (39%) out of 100 cow carcasses and 26 cases (26%) out of 100 studied buffalo carcasses. The findings of this study indicate relatively high infection rate in cattle and buffalo populations of the province. The findings of this study indicate relatively high infection rate in cattle and buffalo populations of the province. So, it is suggested to avoid eating raw and undercooked meat.

کلیدواژه‌ها [English]

  • Sarcocystis
  • Cow
  • Buffalo
  • 18srRNA
  • COX1

Extended Abstract

Introduction

Sarcocytosis is a common disease in humans and domestic animals caused by Sarcocytis species. The evolutionary cycle of this parasite includes the carnivorous host as the definitive host and the herbivorous or omnivorous host as the intermediate host. Cattle are considered intermediate hosts for three Sarcocystis species, including Sarcocystis cruzi, Sarcocystis hirsuta and Sarcocystis homonis. Dogs, cats and primates are definitive hosts of cattle’s Sarcocytis. These three species cause severe symptoms and disease in cattle, symptoms of which include jaundice, myocardial hemorrhage, hair loss, anorexia, weight loss, anemia, decreased milk production, abortion, neurological disorders and death. Five species reported from the buffalo including Sarcocystis fusiformis, Sarcocystis buffalonis, Sarcocystis levini, Sarcocystis dubeyi  and Sarcocystis sciensis. Due to the importance of Sarcocystis as a common zoonotic disease in public health, in this study, the infection rate of Sarcocystis infection and molecular identification of its species by using COX1 and 18srRNA genes in slaughtered cattle and buffaloes of the abattoir of Urmia was investigated.

 

Materials and Methods

For this purpose, samples were taken from the intercostal muscles, diaphragm, esophagus, and thighs of 100 cows and 100 buffaloes slaughtered in the slaughterhouse of Urmia city in 2021. Approximately 50 grams of each of muscles were collected separately in nylon bags and, transferred to Urmia Veterinary University Parasitology Laboratory. Of the 100 buffalo carcasses examined, 49 carcasses belonged to male buffalo and 51 carcasses to female buffalo. Of a total of 100 carcasses examined, 55 carcasses were from buffalo aged three or less than three years (<3) and 45 carcasses were from buffalo aged three or older (>3). In addition, out of a total of 100 cow carcasses examined in this study, 66 carcasses belonged to male cows and 34 carcasses to female cows. Of the total of 100 carcasses examined, 49 carcasses were from cows less than two years old (>2) and 51 carcasses were from cows more than two years old. At first, macroscopic and microscopic evaluation were performed on samples. Then, DNA was extracted from homogenized tissues and molecular investigations were done using COX1 and 18srRNA genes.

 

Discussion

In the macroscopic examination, none of the samples obtained from cows were infected with sarcocystis, while this protozoan was isolated from 11 (11%) samples obtained from buffaloes. The infection rate in the impression smear method in cow and buffaloes samples were 27 (27%) and 16 (16%) and in peptic digestion method was 37 (37%) and 23 (23%), respectively. In parallel, molecular evaluation confirmed the presence of sarcocystis infection in 39 cases (39%) out of 100 cow carcasses and 26 cases (26%) out of 100 studied buffalo carcasses. The results of this study indicate a relatively high rate of infection in the province's cattle and buffalo populations. Therefore, it is recommended to provide control and prevention programs to prevent access to contaminated viscera of dogs and cats in the region and to avoid eating raw and undercooked meat.

Dubey, J. P., Speer, C. & Fayer. R. (1989). Sarcocystosis of Animals and Man. CRC Press, Inc.Boca Raton, 105–145.
Dubey, J. P. & Lindsay, D. S. (2006) Neosporosis, toxoplasmosis, and sarcocystosis in ruminants. The Veterinary clinics of North America Food animal practice. 22, 645-671.
Dubey, J. (2015). Foodborne and Waterborne Zoonotic Sarcocystosis. Journal of Food and Waterborne Parasitology. 1, 2-11.
Daptardar, M., Singh, B. B., Aulakh, R. S. & Gill, J. P. (2016). Prevalence and first molecular identification of Sarcocystis species in cattle and water buffaloes in India. Journal of Acta parasitological, 61(3), 523-528.
Farhang-Pajuh, F., Yakhchali, M. & Mardani, K. (2014). Molecular determination of abundance of infection with Sarcocystis species in slaughtered sheep of Urmia, Iran. Veterinary Research Forum, 5(3), 181-186.
Fayer, R. (2004). Sarcocystis Spp. In Human Infections. Journal of Clinical microbiology reviews, 17, 894-902.
Fayer, R., Esposito, D. H. & Dubey, J. P. (2015). Human Infections with Sarcocystis Species. Journal of Clinical Microbiology Reviews, 28, 295-311.
Gjerde, B. (2016). Molecular characterisation of Sarcocystisbovifelis, Sarcocystisbovini n. sp., Sarcocystis hirsuta and Sarcocystis cruzi from cattle (Bos taurus) and Sarcocystis sinensis from water buffaloes (Bubalus bubalis). Journal of Parasitology research, 115(4), 1473-1492.
Hamidinejat, H., Jalali, M. H. R., Gharibi, D. & Molayan, P. H. (2015). Detection of Sarcocystis spp. in cattle (Bos taurus) and water buffaloes (Bubalus bubalis) in Iran by PCR–RFLP. Journal of Parasitic Diseases, 39(4), 658-662.
Herenda, D. C., P. Chambers & Ettriqui, A. (1994). Manual on Meat Inspection for Developing Countries. Food & Agriculture Org.
Hoeve-Bakker, B., van der Giessen, J. & Franssen, F. (2019). Molecular identification targeting cox1 and 18S genes confirms the high prevalence of Sarcocystis spp. in cattle in the Netherlands. International journal for parasitology, 49(11), 859-866.
Kalantari, N., Khaksar, M., Ghaffari, S. & Hamidekish, S. M. (2016). Molecular analysis of Sarcocystis spp. isolated from sheep (Ovis aries) in Babol area, Mazandaran province, Northern Iran. Iranian journal of parasitology, 11(1),73.
Latif, B., S. Vellayan, C. Heo, M. Kannan Kutty, E. Omar, Abdullah S. & Tappe, D. (2013). High Prevalence of Muscular Sarcocystosis in Cattle and Water Buffaloes from Selangor, Malaysia. Journal of Tropical biomedicine, 30, 699-705.
Mirzaei, M. & Rezaei, H. (2016). A survey on Sarcocystis spp. infection in cattle of Tabriz city, Iran. Journal of parasitic diseases: Journal of official organ of the Indian Society for Parasitology ,40(3), 648-651. (In Persian)
Moré, G., P. Abrahamovich, S. Jurado, D. Bacigalupe, J. Marin, M. Rambeaud, Venturini, L. & Venturini, M. (2011). Prevalence of Sarcocystis Spp. In Argentinean Cattle. Journal of Veterinary parasitology, 177, 162-165.
Moré, G., Schares, S., Maksimov, A., Conraths, F. J., Venturini, M. C. & Schares, G. (2013). Development of a multiplex real time PCR to differentiate Sarcocystis spp. affecting cattle. Journal of Veterinary parasitology, 197(1-2), 85-94.
Moré, G., A. Pantchev, J. Skuballa, M. Langenmayer, P. Maksimov, F. Conraths, Venturini M. C. & Schares, G. (2014). Sarcocystis Sinensis Is the Most Prevalent Thick-Walled Sarcocystis Species in Beef on Sale for Consumers in Germany. Journal of Parasitology Research, 113, 2223-2230.
Najafiyan, H.R., Mohebali, M. & Keshavarz, H. (2008). Study on frequency of sarcocystis spp. by macroscopic and microscopic methods in slaughtered cattle in Shahriar district and their public health importance. Journal of Pajouhesh & Sazandegi, 21(1), 15-19.
 NourollahiFard, S. R., M. Asghari & Nouri, F. (2009). Survey of Sarcocystis infection in slaughtered cattle in Kerman, Iran. Journal of Tropical animal health and production, 41(8),1633-1636. (In Persian)
Razmi, Gh. & Rahbari, S. (2000). Review of Sarcocystis of domestic ruminants in Tehran and Golestan provinces. Scientific Journal of the School of Veterinary Medicine, 3(4), 39-46.
Sarafraz, N., Spotin, A., Haniloo, A. & Fazaeli, A. (2020). Prevalence and molecular analysis of Sarcocystis infections in cattle in Northwest Iran and the first global report of S. gigantea in cattle. Comparative immunology, Journal of microbiology and infectious diseases, 73,101-566. (In Persian)
Shariat Panah, K. (2003). Evaluation of contamination Sarcocysits slaughterhouse in domestic ruminants and tinsel Urmia and its economic importance. Doctor Veterinary Medicine thesis, Faculty of Veterinary Medicine, Islamic Azad University, Urmia branch.
Shekarforosh, S. & Ahmadi, B. (2004). Sarcocytosis Infection in Slaughtered Cattle in Isfahan and Health Care. IBM Journal of Research and Development, 64, 102-4. (In Persian)
Tappe, D., S. Abdullah, C. Heo, Kannan Kutty, M. & Latif, B. (2013). Review Paper Human and Animal Invasive Muscular Sarcocystosis in Malaysia–Recent Cases, Review and Hypotheses. Journal of Tropical biomedicine, 30, 355-366.
Tenter, A. M., Zimmerman, G. L. & Johnson, A. M. (1991). Separation of Antigens from Sarcocystis Species Using Chromatofocusing. The Journal of parasitology, 727-736.