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A long-term preservation method to maintain Toxoplasma gondii in liquid nitrogen



A long-term preservation method to maintain Toxoplasma gondii in liquid nitrogen



Journal of the Egyptian German Society of Zoology. March; 47(D): 67-82



The possible infection of adult BALB/c mice with long-term cryopreserved RH strain of T. gondii in liquid nitrogen was investigated. Tachyzoites of Toxoplasma gondii were cryopreserved at -196deg. C in liquid nitrogen using 10% DMSO. A group of female BALB/C mice were inoculated intraperitoneally by viable tachyzoites. SDS-PAGE and western blot analyses revealed no differences between the polypeptide pattern and reactivity of tachyzoite antigens before and after preservation. However, a decrease in the mean count (6.45 X 106) of tachyzoites was observed after preservation compared with the mean count (8.36 X106) of tachyzoites before preservation but did not reach a significant level (p > 0.05). Also, a nonsignificant (p>0.05) decrease in the mean viability (96%) of tachyzoites was observed after preservation, compared to the mean viability (98%) of tachyzoites before preservation. A group of mice were infected by different volumes (100- to 500 mu L/mouse) of cryopreserved tachyzoites and the survival time was evaluated. The highest survival time (6-8 days) was observed using the dose of 200 mu L per mouse. This dose was used as a primary inoculation dose for subsequent experiments. The thawed tachyzoites were inoculated in a group of mice by the 200 mu L dose. The clinical signs of acute toxoplasmosis were observed in all mice after 6-8 days. An increase in both the mean count and the viability of the tachyzoites exudate (first generation) was observed compared with the mean count and the viability of the preserved tachyzoites. The first generation tachyzoites were inoculated in a group of mice with a dose of 100 mu L /mice containing 7.18 X 106 tachyzoites. The mean tachyzoites count and viability (2nd generation) have been returned to that obtained before preservation. Also, the death time was 2-3 days. This is a reminder that the current scenario is vital not only for this kind of tachyzoites preservation technique to be effective, but also for its sustainability.

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Accession: 037606202

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