Introduction of protocols for mass production of <i>Toxoplasma gondii</i> tachyzoites of the genotype II PRU strain

Bahreini, Mohammad Saleh; Nohtani, Mohammad; Salemi, Amir Masoud; Mirzaeipour, Mehdi; Dastan, Naghmeh; Bajelan, Sara; Asgari, Qasem

doi:10.1002/ame2.12174

Cited by 2 publications

(4 citation statements)

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“…While parasite propagation is mandatory for antigen production, it is also useful for tachyzoites enrichment for typing purposes, and can additionally provide vital information in studies focusing T. gondii lifecycle (and its particular stages), host invasion, host-parasite interactions, and host behaviour [18][19][20][21]. Although some of these goals (e.g., antigen production) may be achieved by using cell-lines for tachyzoites' propagation [9][10][11]13], the animal model remains crucial for studying the natural infection dynamics and parasite dissemination, and is required to confirm findings after initial in vitro investigations [22]. Also, the results of the studies using different cell lines were quite heterogeneous as different approaches were used and, in several cases, a single cell line was evaluated, which hampers the identification of the best cell line for each purpose.…”

Section: Discussionmentioning

confidence: 99%

“…According to the recommendations concerning the animal welfare on behalf of their use in laboratory, we aimed to evaluate the potential of in vitro/in vivo procedures to replace, or at least reduce, the number of sacrificed animals needed for T. gondii propagation in reference laboratories. Some studies have already been performed towards the use of different cell lines for T. gondii propagation, yielding quite heterogeneous results in terms of the obtained parasite concentration and viability [9][10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

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Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model

et al. 2022

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Toxoplasma gondii is an obligate intracellular protozoan. In pregnant women, it can lead to severe birth defects or intrauterine death of the fetus. Most of what is currently know on cell biology of T. gondii comes from studies relying on the RH strain propagated in mice. According to the recommendations concerning the animal welfare, we assayed in vitro/in vivo procedures to replace, or at least reduce, the demanding animal model for strain propagation. We evaluated the genetic and phenotypic stability of the RH strain throughout its parallel continuous propagation in mice, in human foreskin fibroblasts (HFF) and in an alternate fashion of these two procedures. We also assessed the virulence impact on the RH strain after different periods of its long-term propagation strictly in cells. The RH strain completely lost its virulence after long-term passage in HFF. Nevertheless, we obtained a successful outcome with the alternate passaging of the parasite in HFF and in mice as this approach enabled T. gondii to maintain the evaluated phenotypic properties, mainly its virulence potential. Also, no genetic changes were observed in genes known to be highly polymorphic or involved in pathoadaptation. In conclusion, the alternate model seems to be a feasible method for T. gondii propagation and maintenance, strongly impacting the number of sacrificed mice.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model

et al. 2022

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“…Then, a longitudinal incision was made in the abdomen on the skin, the skins were removed, the peritoneal area was flushed with normal saline, the most tachyzoites were collected from the peritoneal area. Based on previous study the yielded tachyzoites were separated and purified [ 18 ].…”

Section: Methodsmentioning

confidence: 99%

“…Tachyzoite of RH strain collected from the peritoneum of infected mice were separated by different centrifugation cycles and washed with PBS [ 18 ]. Then, 10 5 tachyzoites were exposed to various concentrations of compounds (4, 16, 64, 265, 1024 μg/ml) for 1.5 h and 100 μL of each concentration was injected IP to 10 BALB/c mice.…”

Section: Methodsmentioning

confidence: 99%

Synthesis and anti- Toxoplasma activity of indole-triazole compounds on tachyzoites of RH strain

Bahreini

Iraji

Monfared

et al. 2022

Annals of Medicine &Amp; Surgery

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Background Conventional treatment for toxoplasmosis have severe side effects and the inability to completely eradicate the disease. Therefore, the acquisition of new anti- Toxoplasma drugs has always been of interest among researchers. In the present study, we prepare a new indole-triazole derivatives and evaluated their potential anti-parasitic activity against tachyzoites of Toxoplasma RH strain. Materials and methods In this study, after synthesis of the two new compounds of indole-triazole, the effect of their different concentrations (2–1024 μg/ml) were determined on Toxoplasma tachyzoites using flow cytometry. Furthermore, tachyzoites were exposed to different concentrations of compounds (4, 16, 64, 265, 1024 μg/ml) for 1.5 h and their infectivity were evaluated in BALB/c mice. Results The flow cytometry results indicated the benzyl derivative of indole-triazole in various concentrations had a lethal effect on tachyzoites between 11.93% and 89.66%, while the naphthalene derivative had a lethality of 26.63%–66.82%. The infectivity analysis showed that the survival time of mice at concentrations of 1024 μg/ml and 512 μg/ml of benzyl derivatives was significantly increased ( P = 0.008 and P = 0.016, respectively), compared to that in the negative control group. Furthermore, survival time of mice was statistically significant at the concentration of 1024 μg/ml for naphthyl derivative ( P = 0.012). Conclusion Findings of the current study suggested indole triazole compounds, based on their structure and enzymes targeting, have a considerable effect on tachyzoites of T. gondii RH strain and can be considered as a new anti- Toxoplasma agent.

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Introduction of protocols for mass production of Toxoplasma gondii tachyzoites of the genotype II PRU strain

Abstract: This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Cited by 2 publications

References 22 publications

Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model

Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model

Synthesis and anti- Toxoplasma activity of indole-triazole compounds on tachyzoites of RH strain

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