The First Survey of Isolation and Molecular Typing of Toxoplasma gondii by Bioassay and PCR Method in BALB/c Mice in Camels (Camelus dromedarius) from Eastern Iran
-
Amir TAVAKOLI KARESHK
,
Razieh TAVAKOLI OLIAEE
,
Hossein MAHMOUDVAND
,
Amir KEYHANI
,
Mohammad Ali MOHAMMADI
,
Mehdi BAMOROVAT
,
Mohammad Ali HAJHOSSEINI
,
Naser ZIA-ALI
Abstract
Background:This study was designed to isolation of Toxoplasma gondii from camels by bioassay method in mice model and detect parasitic DNA in brain mice by molecular methods.
Methods:A total of 100 tissue samples including heart(n=50),and diaphragm(n=50) were obtained from camels(n=50) slaughtered in abattoirs from three provinces located in eastern Iran.Bioassay method was done in positive MAT camels and Nested PCR performed in tissue samples to amplify the B1 and GRA6 genes. The existence of polymorphic restriction sites for endonuclease MseI was used with PCR-RFLP method and Sequencing analysis to evaluate the prevalence of type strains (I, II and III)
Results: Totally,13(26 %) tissue samples of camels were found positive for the T. gondii B1 gene, including 7(14%) diaphragm,6(12%) heart. Moreover,3(6%) tissue samples of camels were found positive with GRA6 gene for T. gondii. There are three genotypes and mix genotype using MseI enzyme among all positive samples.
Conclusion: The obtained results for the first time demonstrated the presence of T. gondii DNA in the tissues of camels from east of Iran. Since consumption of meat camels are raising in Iran, there may be a high risk of contamination through consumption of products from these hosts due to their susceptibility to the infection.
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2. Gangneux F, Darde ML. Epidemiology of and diagnostic strategies for toxoplasmosis. Clin Microbiol Rev. 2012; 25: 264–296.
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9. Navidpour S, Hoghooghi-Rad N. Seroprevalence of anti-Toxoplasma gondii antibodies in buffaloes in Khuzestan province, Iran. Vet Parasitol. 1998; 77(2-3):191–4.
10. 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: 183-187.
11. Ghazaei C. Serological survey of antibodies to Toxoplasma gondii. Afr J Health Sci. 2006; 13:131–134.
12. Quan JH, Kim TY, Choi IU, Lee YH. Genotyping of a Korean isolate of Toxoplasma gondii by multilocus PCR-RFLP and microsatellite analysis. Korean J Parasitol. 2008; 46(2):105-8.
13. Desmonts G, Naot Y, Remington JS. Immunoglobulin M-immunosorbent agglutination assay for diagnosis of infectious diseases: diagnosis of acute congenital and acquired Toxoplasma infections. J Clin Microbiol. 1981; 14:486–91.
14. Pena HF, Gennari SM, Dubey JP, Su C. Population structure and mouse virulence of Toxoplasma gondii in Brazil. Int J Parasitol.2008; 38:561–569.
15. Fritz H, Barr B, Packham A et al. Methods to produce and safely work with large numbers of Toxoplasma gondii oocysts and bradyzoitecysts. J Microbiol Methods. 2012; 88(1):47–52.
16. Gutierrez J, O'Donovan J, Williams E et al. Detection and quantification of Toxoplasma gondii in ovine maternal and foetal tissues from experimentally infected pregnant ewes using real-time PCR. Vet Parasitol. 2010; 172:8–15.
17. Su C, Zhang X, Dubey JP. Genotyping of Toxoplasma gondii by multilocus PCR-RFLP markers, a high resolution and simple method for identification of parasites. Int J Parasitol.2006;36(7):841–8.
18. Chiabchalard R, Wiengcharoen JT, Sukthana Y. Sensitivity and specificity of PCR for the detection of Toxoplasma gondii and added to laboratory samples. Southeast Asian J Trop Med Public Health.2005; 36:408–411.
19. Kong QM, Shao-Hong LU, Tong QB et al. Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice. Parasites Vectors. 2012; 5: 2.
20. lecordier L, Moleon-Borodowsky I, Dubremetz JF et al. Characterization of a dense granule antigen of Toxoplasma gondii (GRA6) associated to the network of the parasitophorous vacuole. Mol Biochem Parasitol.1995; 70:85-94.
21. Fazaeli A, Carter PE, Darde ML, Pennington TH. Molecular typing of Toxoplasma gondii strains by GRA6 gene sequence analysis. Int J Parasitol.2000; 30(5):637-42.
22. Danehchin L, Razmi Gh, NaghibiA. Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran.Korean J Parasitol. 2016; 54(1):15-20.
23. Tavassoli M, Esmaeilnejad B, Malekifard F et al. Detection of Toxoplasma gondii DNA in Sheep and Goat Milk in Northwest of Iran by PCR-RFLP. Jundishapur J Microbiol. 2013; 6(10): e8201.
24. Khamesipour F, Doosti A, Iranpour Mobarakeh H, Komba EV. Toxoplasma gondii in Cattle, Camels and Sheep in Isfahan and Chaharmahal va Bakhtiary Provinces, Iran. Jundishapur J Microbiol. 2014; 7(6):e17460.
25. Azizi H, Shiran B, Boroujeni AB, Jafari M. Molecular Survey of Toxoplasma gondii in Sheep, Cattle and Meat Products in ChaharmahalvaBakhtiari Province, Southwest of Iran. Iran J Parasitol. 2014; 9(3):429-34.
26. Shaapan RM, Fathia Khalil AM. Evaluation of different Toxoplasma gondii isolates as antigens used in the modified agglutination test for the detection of toxoplasmosis in camels and donkeys. Am-Eurasian J Agr Environ Sci.2008 ;(3):837–841.
27. Dehkordi FS, Borujeni MR, Rahimi E, Abdizadeh R. Detection of Toxoplasma gondii in raw caprine, ovine, buffalo, bovine, and camel milk using cell cultivation, catbioassay, capture ELISA, and PCR methods in Iran. Foodborne Pathog Dis.2013; 10:120–125.
28. Utuk AE, Kirbas A, Babur C, BalkayaI. Detection of Toxoplasma gondiiantibodies and some helminthic parasites in camels from Nevsehir province of Turkey. Isr J Vet Med. 2012;(67):106–108.
29. Gebremedhin EZ, HasenAwel Y, Gebregergs T et al. First report of Toxoplasma gondii in camels(Camelus dromedarius)in Ethiopia: bioassay and seroepidemiological investigation. BMC Vet Res. 2014; 10:222.
30. Tavakoli Kareshk A, Keyhani A, Asadi A et al. Seroprevalence of Toxoplasma gondii infection among childbearing age women in Kerman city, southeastern Iran. J Parasit Dis. 2016;40(4):1544-1547.
31. 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. Tropical Biomedicine .2017; 34(3): 681–690.
32. Switaj K, Master A, Skrzypczak M, Zaborowski P. Recent trends in molecular diagnostics for Toxoplasma gondii infections. Clin Microbiol Infect. 2005; 11:170–6.
33. Zia-Ali N, Fazaeli A, Khoramizadeh M et al. Isolation and molecular characterization of Toxoplasma gondii strains from different hosts in Iran. Parasitol Res.2007;101(1):111-5.
34. Habibi GR, Imani AR, Gholami MR et al. Detection and Identification of Toxoplasma gondii Type One Infection in Sheep Aborted Fetuses in Qazvin Province of Iran. Iran J Parasitol.2012; 7(3): 64-72.
2. Gangneux F, Darde ML. Epidemiology of and diagnostic strategies for toxoplasmosis. Clin Microbiol Rev. 2012; 25: 264–296.
3. Sukthana Y. Toxoplasmosis: beyond animals to humans. Trends Parasitol. 2006; 22: 137–142.
4. Dubey JP. Toxoplasmosis of Animals and Humans. 2nd ed. USA, Florida: CRC Press, Boca Raton. 2009; pp:1-313.
5. Jittapalapong S, Sangvaranond A, Pinyopanuwat N et al. Seroprevalence of Toxoplasma gondii infection in domestic goats in Satun Province, Thailand. Vet Parasitol. 2005; 127(1):17-22.
6. Ghorbani M, Hafizi A, Shegerfcar MT et al. Animal toxoplasmosis in Iran. J Trop Med Hyg.1983; 86(2):73–6.
7. Hoghooghi-Rad N, Afraa M. Prevalence of toxoplasmosis in humans and domestic animals in Ahwaz, capital of Khuzestan Province, south-west Iran. J Trop Med Hyg. 1993;96(3):163–8.
8. Hashemi-Fesharki R. Seroprevalence of Toxoplasma gondii in cattle, sheep and goats in Iran. Vet Parasitol. 1996; 61(1-2):1–3.
9. Navidpour S, Hoghooghi-Rad N. Seroprevalence of anti-Toxoplasma gondii antibodies in buffaloes in Khuzestan province, Iran. Vet Parasitol. 1998; 77(2-3):191–4.
10. 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: 183-187.
11. Ghazaei C. Serological survey of antibodies to Toxoplasma gondii. Afr J Health Sci. 2006; 13:131–134.
12. Quan JH, Kim TY, Choi IU, Lee YH. Genotyping of a Korean isolate of Toxoplasma gondii by multilocus PCR-RFLP and microsatellite analysis. Korean J Parasitol. 2008; 46(2):105-8.
13. Desmonts G, Naot Y, Remington JS. Immunoglobulin M-immunosorbent agglutination assay for diagnosis of infectious diseases: diagnosis of acute congenital and acquired Toxoplasma infections. J Clin Microbiol. 1981; 14:486–91.
14. Pena HF, Gennari SM, Dubey JP, Su C. Population structure and mouse virulence of Toxoplasma gondii in Brazil. Int J Parasitol.2008; 38:561–569.
15. Fritz H, Barr B, Packham A et al. Methods to produce and safely work with large numbers of Toxoplasma gondii oocysts and bradyzoitecysts. J Microbiol Methods. 2012; 88(1):47–52.
16. Gutierrez J, O'Donovan J, Williams E et al. Detection and quantification of Toxoplasma gondii in ovine maternal and foetal tissues from experimentally infected pregnant ewes using real-time PCR. Vet Parasitol. 2010; 172:8–15.
17. Su C, Zhang X, Dubey JP. Genotyping of Toxoplasma gondii by multilocus PCR-RFLP markers, a high resolution and simple method for identification of parasites. Int J Parasitol.2006;36(7):841–8.
18. Chiabchalard R, Wiengcharoen JT, Sukthana Y. Sensitivity and specificity of PCR for the detection of Toxoplasma gondii and added to laboratory samples. Southeast Asian J Trop Med Public Health.2005; 36:408–411.
19. Kong QM, Shao-Hong LU, Tong QB et al. Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice. Parasites Vectors. 2012; 5: 2.
20. lecordier L, Moleon-Borodowsky I, Dubremetz JF et al. Characterization of a dense granule antigen of Toxoplasma gondii (GRA6) associated to the network of the parasitophorous vacuole. Mol Biochem Parasitol.1995; 70:85-94.
21. Fazaeli A, Carter PE, Darde ML, Pennington TH. Molecular typing of Toxoplasma gondii strains by GRA6 gene sequence analysis. Int J Parasitol.2000; 30(5):637-42.
22. Danehchin L, Razmi Gh, NaghibiA. Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran.Korean J Parasitol. 2016; 54(1):15-20.
23. Tavassoli M, Esmaeilnejad B, Malekifard F et al. Detection of Toxoplasma gondii DNA in Sheep and Goat Milk in Northwest of Iran by PCR-RFLP. Jundishapur J Microbiol. 2013; 6(10): e8201.
24. Khamesipour F, Doosti A, Iranpour Mobarakeh H, Komba EV. Toxoplasma gondii in Cattle, Camels and Sheep in Isfahan and Chaharmahal va Bakhtiary Provinces, Iran. Jundishapur J Microbiol. 2014; 7(6):e17460.
25. Azizi H, Shiran B, Boroujeni AB, Jafari M. Molecular Survey of Toxoplasma gondii in Sheep, Cattle and Meat Products in ChaharmahalvaBakhtiari Province, Southwest of Iran. Iran J Parasitol. 2014; 9(3):429-34.
26. Shaapan RM, Fathia Khalil AM. Evaluation of different Toxoplasma gondii isolates as antigens used in the modified agglutination test for the detection of toxoplasmosis in camels and donkeys. Am-Eurasian J Agr Environ Sci.2008 ;(3):837–841.
27. Dehkordi FS, Borujeni MR, Rahimi E, Abdizadeh R. Detection of Toxoplasma gondii in raw caprine, ovine, buffalo, bovine, and camel milk using cell cultivation, catbioassay, capture ELISA, and PCR methods in Iran. Foodborne Pathog Dis.2013; 10:120–125.
28. Utuk AE, Kirbas A, Babur C, BalkayaI. Detection of Toxoplasma gondiiantibodies and some helminthic parasites in camels from Nevsehir province of Turkey. Isr J Vet Med. 2012;(67):106–108.
29. Gebremedhin EZ, HasenAwel Y, Gebregergs T et al. First report of Toxoplasma gondii in camels(Camelus dromedarius)in Ethiopia: bioassay and seroepidemiological investigation. BMC Vet Res. 2014; 10:222.
30. Tavakoli Kareshk A, Keyhani A, Asadi A et al. Seroprevalence of Toxoplasma gondii infection among childbearing age women in Kerman city, southeastern Iran. J Parasit Dis. 2016;40(4):1544-1547.
31. 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. Tropical Biomedicine .2017; 34(3): 681–690.
32. Switaj K, Master A, Skrzypczak M, Zaborowski P. Recent trends in molecular diagnostics for Toxoplasma gondii infections. Clin Microbiol Infect. 2005; 11:170–6.
33. Zia-Ali N, Fazaeli A, Khoramizadeh M et al. Isolation and molecular characterization of Toxoplasma gondii strains from different hosts in Iran. Parasitol Res.2007;101(1):111-5.
34. Habibi GR, Imani AR, Gholami MR et al. Detection and Identification of Toxoplasma gondii Type One Infection in Sheep Aborted Fetuses in Qazvin Province of Iran. Iran J Parasitol.2012; 7(3): 64-72.
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| Issue | Vol 13 No 3 (2018) | |
| Section | Original Article(s) | |
| Keywords | ||
| Toxoplasma gondii Isolation Bioassay Genotyping PCR-RFLP | ||
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How to Cite
1.
TAVAKOLI KARESHK A, TAVAKOLI OLIAEE R, MAHMOUDVAND H, KEYHANI A, MOHAMMADI MA, BAMOROVAT M, HAJHOSSEINI MA, ZIA-ALI N. The First Survey of Isolation and Molecular Typing of Toxoplasma gondii by Bioassay and PCR Method in BALB/c Mice in Camels (Camelus dromedarius) from Eastern Iran. Iran J Parasitol. 2018;13(3):382-391.
