Skip to main content

Advertisement

SpringerLink
Log in

Bioassay-based detection of Toxoplasma gondii in free-range chickens from Khorramabad, Iran: comparison of direct microscopy and semi-nested PCR

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

This study aimed to investigate the occurrence of anti-Toxoplasma gondii antibodies in free-range chickens from Khorramabad, western Iran, and also to compare the performance of direct microscopy and semi-nested PCR in mice bioassayed with tissues from seropositive chickens. We investigated 97 serum samples from free-range chickens, using the modified agglutination test (MAT). Tissues from all seropositive chickens (MAT ≥ 1:10) were bioassayed in mice. All inoculated mice were examined by direct microscopy and a semi-nested PCR targeting the 529 bp repeat element (RE) of the parasite. Anti-T. gondii antibodies were detected in 21.6% of chicken sera. Eighteen of 21 (85.7%) seropositive chickens were positive in mouse bioassay using molecular DNA detection. However, biological forms of the parasite were isolated only from 11 (52.3%) seropositive chickens. Compared with semi-nested PCR, the sensitivity of direct microscopy was 62.1%. It can be concluded that although direct microscopy is a rapid and specific method for the detection of T. gondii, it does not detect the parasite in all experimentally infected mice. The low sensitivity of direct microscopy highlights the need for molecular techniques, such as RE-based semi-nested PCR, to increase the sensitivity of the mouse bioassay.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Dubey JP (2010) Toxoplasmosis of animals and humans, 2nd edn. CRC Press, Boca Raton

    Google Scholar 

  2. Dubey JP (2010) Toxoplasma gondii infections in chickens (Gallus domesticus): prevalence, clinical disease, diagnosis and public health significance. Zoonoses Public Health 57:60–73. https://doi.org/10.1111/j.1863-2378.2009.01274.x

    Article  CAS  PubMed  Google Scholar 

  3. Sabri AR, Hassan L, Sharma RSK, Noordin MM (2019) Toxoplasma gondii infection in native village chickens (Gallus domesticus) in Selangor and Melaka, Malaysia. Trop Biomed 36(3):604–609

    Google Scholar 

  4. Dubey JP, Graham DH, Dahl E, Hilali M, El-Ghaysh A, Sreekumar C, Kwok OCH, Shen SK, Lehmann T (2003) Isolation and molecular characterization of Toxoplasma gondii from chickens and ducks from Egypt. Vet Parasitol 114:89–95. https://doi.org/10.1016/S0304-4017(03)00133-X

    Article  CAS  PubMed  Google Scholar 

  5. Liu XC, He Y, Han DG, Zhang ZC, Li K, Wang S, Xu LX, Yan RF, Li XR (2017) Detection of Toxoplasma gondii in chicken and soil of chicken farms in Nanjing region, China. Infect Dis Poverty 6:62. https://doi.org/10.1186/s40249-017-0277-3

    Article  PubMed  PubMed Central  Google Scholar 

  6. Nasr IA, El-Ashram S, Huang S (2018) Seroprevalence and genotyping of Toxoplasma gondii from free-range chickens in Qassim, Saudi Arabia. Jpn J Vet Res 66(2):113–117

    Google Scholar 

  7. Casartelli-Alves L, Boechat VC, Macedo-Couto R, Ferreira LC, Nicolau JL, Neves LB, Millar PR, Vicente RT, Oliveira RVC, Muniz AG, Bonna ICF, Amendoeira MRR, Silva RC, Langoni H, Schubach TMP, Menezes RC (2014) Sensitivity and specificity of serological tests, histopathology and immunohistochemistry for detection of Toxoplasma gondii infection in domestic chickens. Vet Parasitol 204(3–4):346–351. https://doi.org/10.1016/j.vetpar.2014.05.039

    Article  CAS  PubMed  Google Scholar 

  8. de Lima VY, Langoni H, da Silva AV, Pezerico SB, de Castro APB, da Silva RC, Araújo JP Jr (2011) Chlamydophila psittaci and Toxoplasma gondii infection in pigeons (Columba livia) from São Paulo State, Brazil. Vet Parasitol 175:9–14. https://doi.org/10.1016/j.vetpar.2010.10.006

    Article  PubMed  Google Scholar 

  9. Dubey JP, Desmonts G (1987) Serological responses of equids fed Toxoplasma gondii oocysts. Equine Vet J 19:337–339. https://doi.org/10.1111/j.2042-3306.1987.tb01426.x

    Article  CAS  PubMed  Google Scholar 

  10. Holsback L, Pena HFJ, Ragozo A, Lopes EG, Gennari SM, Soares RM (2012) Serologic and molecular diagnostic and bioassay in mice for detection of Toxoplasma gondii in free ranges chickens from Pantanal of Mato Grosso do Sul. Pesqui Vet Bras 32:721–726. https://doi.org/10.1590/S0100-736X2012000800007

    Article  Google Scholar 

  11. Dubey JP (1998) Refinement of pepsin digestion method for isolation of Toxoplasma gondii from infected tissues. Vet Parasitol 74:75–77. https://doi.org/10.1016/S0304-4017(97)00135-0

    Article  CAS  PubMed  Google Scholar 

  12. Bezerra RA, Carvalho FS, Guimarães LA, Rocha DS, Silva FL, Wenceslau AA, Alburquerque GR (2012) Comparison of methods for detection of Toxoplasma gondii in tissues of naturally exposed pigs. Parasitol Res 110:509–514. https://doi.org/10.1007/s00436-011-2514-1

    Article  PubMed  Google Scholar 

  13. Homan WL, Vercammen M, De Braekeleer J, Verschueren H (2000) 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 30:69–75. https://doi.org/10.1016/S0020-7519(99)00170-8

    Article  CAS  PubMed  Google Scholar 

  14. Dubey JP, Hill DE, Jones JL, Hightower AW, Kirkland E, Roberts JM, Marcet PL, Lehmann T, Vianna MCB, Miska K, Sreekumar C, Kwok OCH, Shen SK, Gamble HR (2005) Prevalence of viable Toxoplasma gondii in beef, chicken and pork from retail meat stores in the United States: risk assessment to consumers. J Parasitol 91:1082–1093. https://doi.org/10.1645/GE-683.1

    Article  CAS  PubMed  Google Scholar 

  15. Dubey JP, Lehmann T, Lautner F, Kwok OCH, Gamble HR (2015) Toxoplasmosis in sentinel chickens (Gallus domesticus) in New England farms: seroconversion, distribution of tissue cysts in brain, heart, and skeletal muscle by bioassay in mice and cats. Vet Parasitol 214:55–58. https://doi.org/10.1016/j.vetpar.2015.09.004

    Article  CAS  PubMed  Google Scholar 

  16. Ying Y, Verma SK, Kwok OCH, Alibana F, Mcleod R, Su C, Dubey JP, Pradhan AK (2017) Prevalence and genetic characterization of Toxoplasma gondii in free-range chickens from grocery stores and farms in Maryland, Ohio and Massachusetts, USA. Parasitol Res 116:1591–1595. https://doi.org/10.1007/s00436-017-5420-3

    Article  PubMed  Google Scholar 

  17. Bayarri S, Gracia MJ, Lázaro R, Perez-Arquillué C, Herrera A (2012) Toxoplasma gondii in meat and food safety implications-a review. In: Lorenzo-Morales J (ed) Zoonosis. InTech, pp 229–254.

  18. Tenter AM, Heckeroth AR, Weiss LM (2000) Toxoplasma gondii: from animals to humans. Int J Parasitol 30:1217–1258. https://doi.org/10.1016/S0020-7519(00)00124-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Shokri A, Sharif M, Teshnizi SH, Sarvi S, Rahimi MT, Mizani A, Ahmadpour E, Montazeri M, Daryani A (2017) Birds and poultries toxoplasmosis in Iran: a systematic review and meta-analysis. Asian Pac J Trop Med 10:635–642. https://doi.org/10.1016/j.apjtm.2017.07.013

    Article  PubMed  Google Scholar 

  20. da Silva AV, Langoni H (2001) The detection of Toxoplasma gondii by comparing cytology, histopathology, bioassay in mice, and the polymerase chain reaction (PCR). Vet Parasitol 97:191–198

    Article  Google Scholar 

  21. Opsteegh M, Langelaar M, Sprong H, den Hartog L, De Craeye S, Bokken G, Ajzenberg D, Kijlstra A, van der Giessen J (2010) Direct detection and genotyping of Toxoplasma gondii in meat samples using magnetic capture and PCR. Int J Food Microbiol 139:193–201

    Article  CAS  Google Scholar 

  22. Gardner IA, Greiner M, Dubey JP (2010) Statistical evaluation of test accuracy studies for Toxoplasma gondii in food animal intermediate hosts. Zoonoses Public Health 57:82–94. https://doi.org/10.1111/j.1863-2378.2009.01281.x

    Article  CAS  PubMed  Google Scholar 

  23. Hamidinejat H, Nabavi L, Mayahi M, Ghourbanpoor M, Pourmehdi Borojeni M, NorollahiFard S, Shokrollahi M (2014) Comparison of three diagnostic methods for the detection of Toxoplasma gondii in free range chickens. Trop Biomed 31:507–513

    CAS  PubMed  Google Scholar 

  24. Dubey JP, Venturini MC, Venturini L, Piscopo M, Graham DH, Dahl E, Sreekumar C, Vianna MC, Lehmann T (2003) Isolation and genotyping of Toxoplasma gondii from free-ranging chickens from Argentina. J Parasitol 89:1063–1064. https://doi.org/10.1645/GE-126

    Article  CAS  PubMed  Google Scholar 

  25. Dubey JP, Morales ES, Lehmann T (2004) Isolation and genotyping of Toxoplasma gondii from free-ranging chickens from Mexico. J Parasitol 90:411–413. https://doi.org/10.1645/GE-194R

    Article  CAS  PubMed  Google Scholar 

  26. Dubey JP, Edelhofer R, Marcet P, Vianna MCB, Kwok OCH, Lehmann T (2005) Genetic and biologic characteristics of Toxoplasma gondii infections in free-range chickens from Austria. Vet Parasitol 133:299–306. https://doi.org/10.1016/j.vetpar.2005.06.006

    Article  CAS  PubMed  Google Scholar 

  27. Dubey JP, Patitucci AN, Su C, Sundar N, Kwok OCH, Shen SK (2006) Characterization of Toxoplasma gondii isolates in free range chickens from Chile, South America. Vet Parasitol 140:76–82. https://doi.org/10.1016/j.vetpar.2006.03.023

    Article  CAS  PubMed  Google Scholar 

  28. Dubey JP, Su C, Oliveira J, Morales JA, Bolaños RV, Sundar N, Kwok OCH, Shen SK (2006) Biologic and genetic characteristics of Toxoplasma gondii isolates in free-range chickens from Costa Rica, Central America. Vet Parasitol 139:29–36. https://doi.org/10.1016/j.vetpar.2006.02.031

    Article  CAS  PubMed  Google Scholar 

  29. Dubey JP, Huong LTT, Lawson BWL, Subekti DT, Tassi P, Cabaj W, Sundar N, Velmurugan GV, Kwok OCH, Su C (2008) Seroprevalence and isolation of Toxoplasma gondii from free-range chickens in Ghana, Indonesia, Italy, Poland, and Vietnam. J Parasitol 94:68–71. https://doi.org/10.1645/GE-1362.1

    Article  CAS  PubMed  Google Scholar 

  30. Dubey JP, Laurin E, Kwowk OCH (2016) Validation of the modified agglutination test for the detection of Toxoplasma gondii in free-range chickens by using cat and mouse bioassay. Parasitology 143:314–319. https://doi.org/10.1017/S0031182015001316

    Article  CAS  PubMed  Google Scholar 

  31. Zewdu E, Tessema TS, Tilahun G, Cox E, Goddeeris B, Dorny P (2015) Comparison of agglutination test, microscopy and nPCR for diagnosis of Toxoplasma gondii isolated from sheep and goat of Central Ethiopia. Ethiop Vet J 19(1):117–129. https://doi.org/10.4314/evj.v19i1.6

    Article  Google Scholar 

  32. Sarkari B, Asgari Q, Bagherian N, Ashkani Esfahani S, Kalantari M, Mohammadpour I, Ashrafmansori M, Amerinia M, Sabet Sarvestani F (2014) Molecular and serological evaluation of Toxoplasma gondii infection in reared turkeys in Fars Province. Iran Jundishapur J Microbiol 7(7):e11598

    PubMed  Google Scholar 

Download references

Acknowledgements

This work was financially supported by the Lorestan University. We would like to thank Professor H. Hamidinejat of Shahid Chamran University for his technical assistance.

Author information

Authors and Affiliations

  1. Department of Parasitology, Faculty of Veterinary Medicine, Lorestan University, Khorramabad, Iran

    Zahra Alizadeh-Sarabi, Zahra Pasandideh & Hamidreza Shokrani

  2. Department of Pathology, Faculty of Veterinary Medicine, Lorestan University, Khorramabad, Iran

    Omid Dezfoulian

Authors
  1. Zahra Alizadeh-Sarabi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zahra Pasandideh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hamidreza Shokrani
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Omid Dezfoulian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hamidreza Shokrani.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This research project was approved by the Faculty of Veterinary Medicine, Lorestan University. During all stages of our research, all applicable international, national, and institutional guidelines for the care and use of animals were followed.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Alizadeh-Sarabi, Z., Pasandideh, Z., Shokrani, H. et al. Bioassay-based detection of Toxoplasma gondii in free-range chickens from Khorramabad, Iran: comparison of direct microscopy and semi-nested PCR. Mol Biol Rep 47, 4969–4974 (2020). https://doi.org/10.1007/s11033-020-05539-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-020-05539-8

Keywords

Search

Navigation