Viability of Toxoplasma gondii tachyzoites in different conditions for parasite transportation

Thi Thuy Nguyen; Ketsarin Kamyingkird; Waraphon Phimpraphai; Tawin Inpankaew

doi:10.14202/vetworld.2022.198-204

Viability of Toxoplasma gondii tachyzoites in different conditions for parasite transportation

Vet World. 2022 Jan;15(1):198-204. doi: 10.14202/vetworld.2022.198-204. Epub 2022 Jan 29.

Authors

Thi Thuy Nguyen^{1

2}, Ketsarin Kamyingkird¹, Waraphon Phimpraphai³, Tawin Inpankaew¹

Affiliations

¹ Department of Parasitology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.
² Department of Veterinary Medicine, Faculty of Animal Science and Veterinary Medicine, University of Agriculture and Forestry, Hue University, Hue, Vietnam.
³ Department of Veterinary Public Health, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.

Abstract

Background and aim: Toxoplasma gondii tachyzoite is the infective stage that causes acute infection, leading to severe toxoplasmosis. The tachyzoite stage has been extensively used for several inoculation purposes, including antigen production, immunological studies, nutrition mechanisms, and in vitro drug trials. The use of fresh tachyzoites is required for inoculation in either in vitro or in vivo studies. However, there is a lack of information on preserving live tachyzoites during transportation from laboratories to inoculation sites. Therefore, this study aimed to validate suitable preservative conditions for maintaining live parasites by determining the survival and viability of T. gondii tachyzoites on the basis of different media, temperatures, and incubation times.

Materials and methods: The free live T. gondii tachyzoites were evaluated on their viability when maintained in different media without 5% Carbon dioxide (CO₂). The purified tachyzoites of the RH and PLK strains were individually suspended in normal saline (NS), phosphate-buffered saline (PBS), minimum essential medium (MEM), and MEM with 10% fetal bovine serum (MEM-FBS) and incubated for 6 h at ice-cold (IC; 3-9°C) and room temperature (RT; 25°C). Parasite survival was measured at the 0, 1^st, 2^nd, 3^rd, 4^th, 5^th, and 6^th h post-incubation using the trypan blue exclusion test.

Results: The viability was in the range of 85.0%-91.0% for IC using NS and 81.0%-85.1% (IC) and 75.3%-77.5% (RT) using PBS. The viability was approximately 75.0%-83.0% (IC) and 70.0%-79.0% (RT) using MEM and MEM-FBS. There was a significant difference in the viability between the seven periods on the basis of one-way repeated Analysis of variance and Friedman analyses. Parasite survival slightly reduced (20.0%-30.0%) in NS and MEM-FBS at both temperatures during incubation. Notably, PBS could not support tachyzoite viability after 3 h post-incubation.

Conclusion: NS was a suitable preservative for maintaining purified T. gondii tachyzoites during transportation at IC and RT without 5% CO₂ supplementation. This could be a valuable medium for parasite transportation, especially when there is a large distance between the laboratory and inoculation site.

Keywords: T. gondii tachyzoites; preservation time; temperature; transportation; viability.