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

Mol Biol Rep. 2020 Jul;47(7):4969-4974. doi: 10.1007/s11033-020-05539-8. Epub 2020 Jun 23.

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.

Keywords: Chickens; Iran; Microscopic cyst detection; Mouse bioassay; Toxoplasma gondii.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Chickens / parasitology*
  • DNA, Protozoan / genetics
  • Liquid Biopsy / standards
  • Liquid Biopsy / veterinary
  • Mice
  • Microscopy / standards
  • Microscopy / veterinary
  • Molecular Diagnostic Techniques / standards
  • Molecular Diagnostic Techniques / veterinary
  • Polymerase Chain Reaction / standards
  • Polymerase Chain Reaction / veterinary
  • Poultry Diseases / blood
  • Poultry Diseases / diagnosis*
  • Poultry Diseases / parasitology
  • Repetitive Sequences, Nucleic Acid
  • Toxoplasma / cytology
  • Toxoplasma / genetics
  • Toxoplasma / pathogenicity
  • Toxoplasmosis, Animal / blood
  • Toxoplasmosis, Animal / diagnosis*
  • Toxoplasmosis, Animal / parasitology

Substances

  • DNA, Protozoan