Detection of B1 Gene of Toxoplasma gondii infection in Slaughtered Sheep in Isfahan Using the Nested PCR

Subject Areas : Journal of Animal Biology

Hamid Raieszadeh ¹ , Abdolah Jamshidi ² , Vahid Noaman ³ , Gholamreza Razmi ⁴

1 - Department of Food Hygiene, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran
2 - Department of Food Hygiene, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran
3 - Parasitic Diseases Research Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, IranOrganization (AREEO), Karaj, Iran
4 - Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

Received: 2020-07-19 Accepted : 2020-10-16 Published : 2021-02-19

Keywords: Sheep, Isfahan, Toxoplasma gondii, Nested-PCR,

Abstract :

Toxoplasma gondii is a common pathogen in humans and animals which can infect humans through the food sources. The consumption of mutton is more popular than other sources of animal protein in Iran. Since the sheep heart is usually eaten half-cooked and grilled, the transmission of toxoplasmosis occurs more often through the consumption of sheep heart. On the other hand, because the density of parasitic cysts in the brain and heart tissue is higher than other oral organs, sheep heart samples were used in this study. In the recent study, to detect toxoplasma in sheep heart slaughtered in Isfahan, 250 heart samples were collected from different sexes and ages in different seasons over the course of one year. After extracting the parasite genome, two pairs of primers related to B1 gene were amplified by nested PCR method. The results showed that 78% of the samples were generally infected with toxoplasma. In this study, there was no significant relationship between infection percentage and age and sex, but infection rate in cold seasons was significantly higher than in the warm seasons. Since the factors such as high humidity and low-temperature help to oocysts' survival, long exposure with toxoplasma oocysts can increase the sheep infection rate in the cold season. By considering the importance of this disease in sheep deaths and abortions and the importance of public health, appropriate control programs to prevent this zoonotic disease at the provincial level should be included in veterinary policies.

References:

1. Akhoundi S.,  Youssefi, M.R., 2017. Seroprevalence of sheep toxoplasmosis in north of Iran. Trakia Journal of Sciences, 1: 79-82.

2. Armand B., Solhjoo K., Shabani-Kordshooli M., Davami M.H., Sadeghi M., 2016. Toxoplasma infection in sheep from south of Iran monitored by serological and molecular methods; risk assessment to meat consumers. Veterinary World,9(8): 850-855.

3. Asgari Q., Sarnevesht J., Kalantari M., Sadat S.J.A., Motazedian M.H., Sarkari B., 2011. Molecular survey of Toxoplasma infection in sheep and goat from Fars province, Southern Iran. Tropical Animal Health and production, 43(2): 389-92.

4. Azizi H., Shiran B., Boroujeni A.B., Jafari M., 2014. Molecular Survey of Toxoplasma gondii in Sheep, Cattle and Meat Products in Chaharmahal va Bakhtiari Province, Southwest of Iran. Iranian Journal of Parasitology, 9(3): 429-34.

5. Belluco S., Mancin M., Conficoni D., Simonato G., Pietrobelli M., Ricci A., 2016. Investigating the Determinants of Toxoplasma gondii Prevalence in Meat: A Systematic Review and Meta-Regression. PLoS One, 11(4): 53-56.

6. Chiabchalard R., Wiengcharoen JT., Sukthana Y., 2005. Sensitivity and specificity of polymerase chain reaction for the detection of Toxoplasma gondii DNA added to laboratory samples. Southeast Asian Journal of Tropical Medicine and Public Health, 36(2): 408-11.

7. Daryani A., Sarvi S., Aarabi M., Mizani A., Ahmadpour E., Shokri A., 2014. Seroprevalence of Toxoplasma gondii in the Iranian general population: a systematic review and meta-analysis. Acta Tropica, 137: 185-94.

8. Dubey, J.P., 2010. Toxoplasmosis of Animals and Humans, 2nd Edn. Boca Raton, FL: CRC Press; Taylor & Francis Group.

9. Dubey J., 2009.Toxoplasmosis in sheep—the last 20 years. Veterinary Parasitology, 163(1): 1-14.

10. Dubey JP. 2009. Toxoplasmosis of animals and humans: CRC press.

11. Dubey J.P., Beattie C., 1988. Toxoplasmosis of animals and man: CRC Press, Inc.

12. Fallahi S., Tabaei S.J., Pournia Y., Zebardast N., Kazemi B., 2014. Comparison of loop-mediated isothermal amplification (LAMP) and nested-PCR assay targeting the RE and B1 gene for detection of Toxoplasma gondii in blood samples of children with leukaemia. Diagnostic Microbiology and Infectious Disease, 79(3): 347-54.

13. Hashemi-Fesharki R., 1996. Seroprevalence of Toxoplasma gondii in cattle, sheep and goats in Iran. Veterinary Parasitolology, 61(1-2): 1-3.

14. Ihira M., Akimoto S., Miyake F., Fujita A., Sugata K., Suga S., 2007. Direct detection of human herpesvirus 6 DNA in serum by the loop-mediated isothermal amplification method. Journal of Clinical Virology, 39(1) :22-26.

15. Jones J., Dubey J., 2012. Foodborne toxoplasmosis.Clinical Infectious Diseases, 55(6): 845–851.

16. Kalogianni D.P., Elenis D.S., Christopoulos T.K., Ioannou P.C., 2007. Multiplex quantitative competitive polymerase chain reaction based on a multianalyte hybridization assay performed on spectrally encoded microspheres. Analytical Chemistry, 79(17): 6655-61.

17. Khezri M., Mohammadian B., Esmailnia K., Khezri O., 2012. Toxoplasmosis in sheep from Kurdistan province, Iran. African Journal of Microbiology Research, 6(18): 3989–92.

16. Levine N.D., 1985. Veterinary protozoology: Iowa State University Press Ames.

17. Macpherson C.N., 2005. Human behaviour and the epidemiology of parasitic zoonoses. International journal for parasitology, 35(11-12): 1319-31.

18. Notomi T., Okayama H., Masubuchi H., Yonekawa T., Watanabe K., Amino N., 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Research, 28(12): 63-69.

19. Rahdar M., Samarbaf-Zadeh A., Arab L., 2012. Evaluating the prevalence of Toxoplasma gondii in meat and meat products in Ahvaz by PCR method. Jundishapur Journal of Microbiology, 5(4): 570-3.

20. Rassouli M., Razmi G., Bassami M., Movassaghi A., Azizzadeh M., 2011. Study on ovine abortion associated with Toxoplasma gondii in affected herds of Khorasan Razavi Province, Iran based on PCR detection of fetal brains and maternal serology. Parasitology, 138(06): 691-7.

21. Razmi G.R., Ghezi K., Mahooti A., Naseri Z., 2010. A serological study and subsequent isolation of Toxoplasma gondii from aborted ovine fetuses in Mashhad area, Iranan Journal of Parasitology, 96(4): 812-814.

22. Razmi G.R., 1993. Seroprevalence of toxoplasmosis and its Importance in abortion among ewes in Mazandaran province, Iran: Dissertation. University of Tehran.

23. Roberts L., Kwan I., Evans P., 2002. Does animal experimentation inform human healthcare? Observations from a systematic review of international animal experiments on fluid resuscitation. British Medical Journal, 324(7335): 474–476.

24. Sarvi S., Daryani A., Rahimi M.T., Aarabi M., Shokri A., Ahmadpour E., 2015. Cattle toxoplasmosis in Iran: a systematic review and meta–analysis. Asian Pacific Journal of Tropical Medicine, 8(2): 120-6.

25. Scallan E., Hoekstra R., Mahon B., Jones T., Griffin P., 2015. An assessment of the human health impact of seven leading foodborne pathogens in the United States using disability adjusted life years. Epidemiology and Infection, 143(13): 2795-804.

26. Sharif M., Sarvi S., Shokri A., Teshnizi S.H., Rahimi M., Mizani A., 2015. Toxoplasma gondii infection among sheep and goats in Iran: a systematic review and meta-analysis. Parasitology Research, 114(1):1-16.

27. Suleman E., Mtshali MS., Lane E., 2016. Investigation of false positives associated with loop-mediated isothermal amplification assays for detection of Toxoplasma gondii in archived tissue samples of captive felids. Journal of Veterinary Diagnostic Investigation, 28: 536-542.

28. Tavassoli M., Ghorbanzadehghan M., Esmaeilnejad B., 2013. Detection of Toxoplasma gondii in sheep and goats blood samples by PCR-RFLP in Urmia. Veterinary Reserch Forum, 4: 43-47.

29. Valian H.K., Ebrahimi A., 1994. Prevalence of Toxoplasma gondii in birds of Kerman city by serological and parasitological methods. Iranian Journal of Public Health, 23(1-4): 25-34.

30. Van der Puije W.N.A., Bosompem K.M., Canacoo E.A., Wastling J.M., Akanmori B.D., 2000. The prevalence of anti-Toxoplasma gondii antibodies in Ghanaian sheep and goats. Acta Tropica, 76:21-26