Frequency of Toxoplasma gondii in Livestock Meats Using Nested PCR and RFLP in Sabzevar City, Iran
-
Jalal Aliabadi
,
Amir Tavakoli kareshk
,
Iraj Sharifi
,
Mohammad Ali Mohammadi
,
Arghavan Ziaali
,
Hossein Kamyabi
,
Naser Ziaali
,
Seyed Mohammad Mousavi
Abstract
Background: We investigated the prevalence of Toxoplasma gondii in slaughtered livestock (sheep, goats, camels) in Sabzevar, Eastern Iran using nested PCR methods.
Methods: Samples of diaphragm and heart tissue were collected from 40 sheep, 40 goats, and 40 camels at local slaughterhouses. DNA was extracted from the samples and analyzed using nested PCR targeting the B1 gene of T. gondii.
Results: The results indicate an overall prevalence of 60% (24/40) in sheep, with 37.5% (15/40) of diaphragm samples and 22.5% (9/40) of heart samples testing positive. In goats, the overall prevalence was 52.5% (21/40), with 35% (14/40) of diaphragm and 17.5% (7/40) of heart samples positive. Camels showed the highest prevalence at 65% (26/40), with 45% (18/40) of diaphragm and 20% (8/40) of heart samples infected. Diaphragm tissue showed higher infection rates compared to the heart in all species. No significant difference was found in infection rates between male and female animals. RFLP analysis using XhoI on selected positive PCR products differentiated genotypes based on digestion patterns. Among analyzed samples (19 per species), genotypes II/III were identified in 57.9% (11/19) of sheep, 36.8% (7/19) of goats, and 47.4% (9/19) of camels, with the remainder likely genotype I or undigested II/III.
Conclusion: The high prevalence rates observed, especially in diaphragm tissue and in camels, have important implications for public health and food safety. Enhanced meat inspection, public education, and further research on effective control measures are recommended.
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4. Montoya JG. Systematic screening and treatment of toxoplasmosis during pregnancy: is the glass half full or half empty? Am J Obstet Gynecol. 2018;219(4):315–9.
5. Jones JL, Lopez A, Wilson M, Schulkin J, Gibbs R. Congenital toxoplasmosis: a review. Obstet Gynecol Surv. 2001;56(5):296–305.
6. Innes EA. A brief history and overview of Toxoplasma gondii. Zoonoses Public Health. 2010;57(1):1–7.
7. Dubey JP, Jones JL. Toxoplasma gondii infection in humans and animals in the United States. Int J Parasitol. 2008;38(11):1257–78.
8. Tonouhewa AB, Akpo Y, Sessou P, Adoligbe C, Yessinou E, Hounmanou YG, et al. Toxoplasma gondii infection in meat animals from Africa: Systematic review and meta-analysis of sero-epidemiological studies. Vet World. 2017;10(2):194-208.
9. Sharif M, Daryani A, Nasrolahei M, Ziapour SP. Prevalence of Toxoplasma gondii antibodies in stray cats in Sari, northern Iran. Trop Anim Health Prod.2009;41(2):183–7.
10. Gebremedhin EZ, Abdurahaman M, Tessema TS, et al. Isolation and genotyping of viable Toxoplasma gondii from sheep and goats in Ethiopia destined for human consumption. Parasit Vectors. 2014;7:425.
11. Homan WL, Vercammen M, Braekeleer J, Verschueren H. Identification of a 200-to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. Int J Parasitol. 2000;30(1):69–75.
12. Tavakoli Kareshk A, Mahmoudvand H, Keyhani A, et al. Molecular detection and genetic diversity of Toxoplasma gondii in different tissues of sheep and goat in Eastern Iran. Trop Biomed. 2017;34(3):681–90.
13. El-Nawawi FA, Tawfik MA, Shaapan RM. Methods for inactivation of Toxoplasma gondii cysts in meat and tissues of experimentally infected sheep. Foodborne Pathog Dis. 2008; 5(5):687-90.
14. Bezerra RA, Carvalho FS, Guimarães LA, et al. Comparison of methods for detection of Toxoplasma gondii in tissues of naturally exposed pigs. Parasitol Res.2012; 110(2):509-14.
15. Mahami-Oskouei M, Moradi M, Fallah E, Hamidi F. Molecular detection and genotyping of Toxoplasma gondii in chicken, beef, and lamb meat consumed in Northwestern Iran. Iran J Parasitol.2017;12(1):38–45.
16. Firouzeh N, Borj HF, Ziaali N, Kareshk AT, Ahmadinejad M, Shafiei R. Genetic diversity of Toxoplasma gondii by serological and molecular analyzes in different sheep and goat tissues in northeastern Iran. Iran J Parasitol.2023;18(2):217-223.
17. Afshar-Mohammadian M, Rezadoost MH, Fallah SF. Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit. MethodsX. 2018; 5:352–61.
18. Grigg ME, Boothroyd JC. Rapid identification of virulent type I strains of the protozoan pathogen Toxoplasma gondii by PCR-restriction fragment length polymorphism analysis at the B1 gene. J Clin Microbiol. 2001;39(1):398–400.
19. Hajimohammadi B, Ahmadian S, Firoozi Z, et al. A Meta-Analysis of the Prevalence of Toxoplasmosis in Livestock and Poultry Worldwide. Ecohealth. 2022;19(1):55-74.
20. Gisbert Algaba I, Verhaegen B, Jennes M, et al. Pork as a source of transmission of Toxoplasma gondii to humans: a parasite burden study in pig tissues after infection with different strains of Toxoplasma gondii as a function of time and different parasite stages. Int J Parasitol. 2018;48(7):555-560.
21. Khalil MK, Elrayah IE. Seroprevalence of Toxoplasma gondii antibodies in farm animals (camels, cattle, and sheep) in Sudan. Int J Vet Med Animal Health. 2011;4(3):36–9.
22. Ghoneim NH, Shalaby SI, Hassanain NA, Zeedan GSG, Soliman YA, Abdalhamed AM. Comparative Study Between Serological and Molecular Methods for Diagnosis of Toxoplasmosis in Women and Small Ruminants in Egypt. Foodborne Pathog Dis. 2010;7(1):17–22.
23. Rodriguez-Fernandez V, Bruschi F. Toxoplasma gondii in Marine Life of Italian Coasts, What Do We Know So Far? Parasitologia. 2023;3(4):364–73.
24. Zia-Ali N, Fazaeli A, Khoramizadeh M, Ajzenberg D, Dardé M, Keshavarz-Valian H. Isolation and molecular characterization of Toxoplasma gondii strains from different hosts in Iran. Parasitol Res. 2007;101(1):111–5.
25. Asgari Q, Sarnevesht J, Kalantari M, Sadat SJA, Motazedian MH, Sarkari B. Molecular survey of Toxoplasma infection in sheep and goat from Fars province, Southern Iran. Trop Anim Health Prod. 2011; 43(2):389–92.
26. Abdelgadier A, Assaad N, Elhussein Z, et al. Prevalence of Toxoplasma gondii infection in animals of the Arabian Peninsula between 2000-2020: A systematic review and meta-analysis. Vet Med Sci. 2023; 9(1):471–80.
27. Atif I, Touloun O, Boussaa S. Toxoplasma gondii in humans, animals and in the environment in Morocco: a literature review. Gut Pathog. 2024;16(1):53.
28. Juránková J, Basso W, Neumayerová H, et al. Brain is the predilection site of Toxoplasma gondii in experimentally inoculated pigs as revealed by magnetic capture and real-time PCR. Food Microbiol. 2014; 38:167–70.
29. Khames M, Yekkour F, Fernández-Rubio C, Aubert D, Nguewa P, Villena I. Serological survey of cattle toxoplasmosis in Medea, Algeria. Vet Parasitol Reg Stud Reports.2018; 12:89–90.
30. Ramzan M, Akhtar M, Muhammad F, et al. Seroprevalence of Toxoplasma gondii in sheep and goats in Rahim Yar Khan (Punjab), Pakistan. Trop Anim Health Prod. 2009; 41(7):1225–9.
31. Dubey JP. Toxoplasmosis in sheep—The last 20 years Veterinary. Vet Parasitol. 2009;163(1-2):1-14.
32. Gilot-Fromont E, Lélu M, Dardé ML, et al. The life cycle of Toxoplasma gondii in the natural environment. Toxoplasmosis-Recent Advances. 2012; 10:2845.
33. Kijlstra A, Jongert E. Control of the risk of human toxoplasmosis transmitted by meat. Int J Parasitol. 2008;38(12):1359–70.
34. Vitale M, Galluzzo P, Currò V, Gozdzik K, Schillaci D, Di Marco Lo Presti V. A high sensitive nested PCR for Toxoplasma gondii detection in animal and food samples. J Microb Biochem Technol. 2013;5(02):39–41.
35. Hill D, Dubey JP. Toxoplasma gondii: transmission, diagnosis and prevention. Clin Microbiol Infect. 2002;8(10):634–40.
36. Howe DK, Sibley LD. Toxoplasma gondii comprises three clonal lineages: correlation of parasite genotype with human disease. J Infect Dis. 1995;172(6):1561–6.
37. Behnke MS, Khan A, Wootton JC, Dubey JP, Tang K, Sibley LD. Virulence differences in Toxoplasma mediated by amplification of a family of polymorphic pseudokinases. Proc Natl Acad Sci U S A. 2011 ;108(23):9631–6.
38. Fernández-Escobar M, Calero-Bernal R, Regidor-Cerrillo J, et al. Isolation, Genotyping, and Mouse Virulence Characterization of Toxoplasma gondii From Free Ranging Iberian Pigs. Front Vet Sci. 2020; 7:604782.
39. Sánchez-Sánchez R, Ferre I, Regidor-Cerrillo J, Gutiérrez-Expósito D, et al. Virulence in Mice of a Toxoplasma gondii Type II Isolate Does Not Correlate with the Outcome of Experimental Infection in Pregnant Sheep. Front Cell Infect Microbiol. 2019; 8:436.
40. Gorgani-Firouzjaee T, Kalantari N, Ghaffari S. Molecular identification and genotyping of Toxoplasma gondii isolated from sheep and cattle in northern Iran. Vet Res Forum. 2022; 13(3):371–8.
2. Webster JP. Dubey, J.P. Toxoplasmosis of Animals and Humans. Parasites & Vectors. 2010 Nov 23;3(1):1–2.
3. Kanani B, Namaei MH, Kareshk AT, Solgi R. Seroprevalence of Toxoplasma gondii infection among women of reproductive age in Birjand, Iran. Mod Care J. 2022;19(4): e127872.
4. Montoya JG. Systematic screening and treatment of toxoplasmosis during pregnancy: is the glass half full or half empty? Am J Obstet Gynecol. 2018;219(4):315–9.
5. Jones JL, Lopez A, Wilson M, Schulkin J, Gibbs R. Congenital toxoplasmosis: a review. Obstet Gynecol Surv. 2001;56(5):296–305.
6. Innes EA. A brief history and overview of Toxoplasma gondii. Zoonoses Public Health. 2010;57(1):1–7.
7. Dubey JP, Jones JL. Toxoplasma gondii infection in humans and animals in the United States. Int J Parasitol. 2008;38(11):1257–78.
8. Tonouhewa AB, Akpo Y, Sessou P, Adoligbe C, Yessinou E, Hounmanou YG, et al. Toxoplasma gondii infection in meat animals from Africa: Systematic review and meta-analysis of sero-epidemiological studies. Vet World. 2017;10(2):194-208.
9. Sharif M, Daryani A, Nasrolahei M, Ziapour SP. Prevalence of Toxoplasma gondii antibodies in stray cats in Sari, northern Iran. Trop Anim Health Prod.2009;41(2):183–7.
10. Gebremedhin EZ, Abdurahaman M, Tessema TS, et al. Isolation and genotyping of viable Toxoplasma gondii from sheep and goats in Ethiopia destined for human consumption. Parasit Vectors. 2014;7:425.
11. Homan WL, Vercammen M, Braekeleer J, Verschueren H. Identification of a 200-to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. Int J Parasitol. 2000;30(1):69–75.
12. Tavakoli Kareshk A, Mahmoudvand H, Keyhani A, et al. Molecular detection and genetic diversity of Toxoplasma gondii in different tissues of sheep and goat in Eastern Iran. Trop Biomed. 2017;34(3):681–90.
13. El-Nawawi FA, Tawfik MA, Shaapan RM. Methods for inactivation of Toxoplasma gondii cysts in meat and tissues of experimentally infected sheep. Foodborne Pathog Dis. 2008; 5(5):687-90.
14. Bezerra RA, Carvalho FS, Guimarães LA, et al. Comparison of methods for detection of Toxoplasma gondii in tissues of naturally exposed pigs. Parasitol Res.2012; 110(2):509-14.
15. Mahami-Oskouei M, Moradi M, Fallah E, Hamidi F. Molecular detection and genotyping of Toxoplasma gondii in chicken, beef, and lamb meat consumed in Northwestern Iran. Iran J Parasitol.2017;12(1):38–45.
16. Firouzeh N, Borj HF, Ziaali N, Kareshk AT, Ahmadinejad M, Shafiei R. Genetic diversity of Toxoplasma gondii by serological and molecular analyzes in different sheep and goat tissues in northeastern Iran. Iran J Parasitol.2023;18(2):217-223.
17. Afshar-Mohammadian M, Rezadoost MH, Fallah SF. Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit. MethodsX. 2018; 5:352–61.
18. Grigg ME, Boothroyd JC. Rapid identification of virulent type I strains of the protozoan pathogen Toxoplasma gondii by PCR-restriction fragment length polymorphism analysis at the B1 gene. J Clin Microbiol. 2001;39(1):398–400.
19. Hajimohammadi B, Ahmadian S, Firoozi Z, et al. A Meta-Analysis of the Prevalence of Toxoplasmosis in Livestock and Poultry Worldwide. Ecohealth. 2022;19(1):55-74.
20. Gisbert Algaba I, Verhaegen B, Jennes M, et al. Pork as a source of transmission of Toxoplasma gondii to humans: a parasite burden study in pig tissues after infection with different strains of Toxoplasma gondii as a function of time and different parasite stages. Int J Parasitol. 2018;48(7):555-560.
21. Khalil MK, Elrayah IE. Seroprevalence of Toxoplasma gondii antibodies in farm animals (camels, cattle, and sheep) in Sudan. Int J Vet Med Animal Health. 2011;4(3):36–9.
22. Ghoneim NH, Shalaby SI, Hassanain NA, Zeedan GSG, Soliman YA, Abdalhamed AM. Comparative Study Between Serological and Molecular Methods for Diagnosis of Toxoplasmosis in Women and Small Ruminants in Egypt. Foodborne Pathog Dis. 2010;7(1):17–22.
23. Rodriguez-Fernandez V, Bruschi F. Toxoplasma gondii in Marine Life of Italian Coasts, What Do We Know So Far? Parasitologia. 2023;3(4):364–73.
24. Zia-Ali N, Fazaeli A, Khoramizadeh M, Ajzenberg D, Dardé M, Keshavarz-Valian H. Isolation and molecular characterization of Toxoplasma gondii strains from different hosts in Iran. Parasitol Res. 2007;101(1):111–5.
25. Asgari Q, Sarnevesht J, Kalantari M, Sadat SJA, Motazedian MH, Sarkari B. Molecular survey of Toxoplasma infection in sheep and goat from Fars province, Southern Iran. Trop Anim Health Prod. 2011; 43(2):389–92.
26. Abdelgadier A, Assaad N, Elhussein Z, et al. Prevalence of Toxoplasma gondii infection in animals of the Arabian Peninsula between 2000-2020: A systematic review and meta-analysis. Vet Med Sci. 2023; 9(1):471–80.
27. Atif I, Touloun O, Boussaa S. Toxoplasma gondii in humans, animals and in the environment in Morocco: a literature review. Gut Pathog. 2024;16(1):53.
28. Juránková J, Basso W, Neumayerová H, et al. Brain is the predilection site of Toxoplasma gondii in experimentally inoculated pigs as revealed by magnetic capture and real-time PCR. Food Microbiol. 2014; 38:167–70.
29. Khames M, Yekkour F, Fernández-Rubio C, Aubert D, Nguewa P, Villena I. Serological survey of cattle toxoplasmosis in Medea, Algeria. Vet Parasitol Reg Stud Reports.2018; 12:89–90.
30. Ramzan M, Akhtar M, Muhammad F, et al. Seroprevalence of Toxoplasma gondii in sheep and goats in Rahim Yar Khan (Punjab), Pakistan. Trop Anim Health Prod. 2009; 41(7):1225–9.
31. Dubey JP. Toxoplasmosis in sheep—The last 20 years Veterinary. Vet Parasitol. 2009;163(1-2):1-14.
32. Gilot-Fromont E, Lélu M, Dardé ML, et al. The life cycle of Toxoplasma gondii in the natural environment. Toxoplasmosis-Recent Advances. 2012; 10:2845.
33. Kijlstra A, Jongert E. Control of the risk of human toxoplasmosis transmitted by meat. Int J Parasitol. 2008;38(12):1359–70.
34. Vitale M, Galluzzo P, Currò V, Gozdzik K, Schillaci D, Di Marco Lo Presti V. A high sensitive nested PCR for Toxoplasma gondii detection in animal and food samples. J Microb Biochem Technol. 2013;5(02):39–41.
35. Hill D, Dubey JP. Toxoplasma gondii: transmission, diagnosis and prevention. Clin Microbiol Infect. 2002;8(10):634–40.
36. Howe DK, Sibley LD. Toxoplasma gondii comprises three clonal lineages: correlation of parasite genotype with human disease. J Infect Dis. 1995;172(6):1561–6.
37. Behnke MS, Khan A, Wootton JC, Dubey JP, Tang K, Sibley LD. Virulence differences in Toxoplasma mediated by amplification of a family of polymorphic pseudokinases. Proc Natl Acad Sci U S A. 2011 ;108(23):9631–6.
38. Fernández-Escobar M, Calero-Bernal R, Regidor-Cerrillo J, et al. Isolation, Genotyping, and Mouse Virulence Characterization of Toxoplasma gondii From Free Ranging Iberian Pigs. Front Vet Sci. 2020; 7:604782.
39. Sánchez-Sánchez R, Ferre I, Regidor-Cerrillo J, Gutiérrez-Expósito D, et al. Virulence in Mice of a Toxoplasma gondii Type II Isolate Does Not Correlate with the Outcome of Experimental Infection in Pregnant Sheep. Front Cell Infect Microbiol. 2019; 8:436.
40. Gorgani-Firouzjaee T, Kalantari N, Ghaffari S. Molecular identification and genotyping of Toxoplasma gondii isolated from sheep and cattle in northern Iran. Vet Res Forum. 2022; 13(3):371–8.
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| Issue | Vol 21 No 2 (2026) | |
| Section | Original Article(s) | |
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| Toxoplasma gondii Meat Parasitology Iran | ||
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How to Cite
1.
Aliabadi J, Tavakoli kareshk A, Sharifi I, Mohammadi MA, Ziaali A, Kamyabi H, Ziaali N, Mousavi SM. Frequency of Toxoplasma gondii in Livestock Meats Using Nested PCR and RFLP in Sabzevar City, Iran. Iran J Parasitol. 2026;21(2):237-246.
