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Comparison of Eight Cell-Free Media for Maintenance of Toxoplasma gondii Tachyzoites.

Kalani H, Daryani A, Sharif M, Ahmadpour E, Alizadeh A, Nasrolahei M, Sarvi S, Kalani F, Faridnia R - Iran J Parasitol (2016 Jan-Mar)

Bottom Line: The survival rate and viability assessment of parasites were performed daily and the results were analyzed using Univariate tests.Tachyzoites survival rate in PBS (4 °C) and normal saline (4 °C) were considerably high, compared to other solutions in different conditions (P<0.001).The best temperature for Toxoplasma maintenance was 4 °C (P<0.001).

View Article: PubMed Central - PubMed

Affiliation: Toxoplasmosis Research Center, Mazandaran University of Medical Sciences, Sari, Iran.

ABSTRACT

Background: Toxoplasmosis is considered as one of the most common infectious diseases caused by the protozoan parasite Toxoplasma gondii. Tachyzoite is the main form of Toxoplasma and continuously is maintained in cell culture or injected into the mice peritoneal cavity. This study was designed to evaluate the survival rate of RH strain of T. gondii tachyzoites in different cell free, nutrient and biological media at different temperatures.

Methods: This experimental study was performed at the Toxoplasmosis Research Center, Mazandaran University of Medical Sciences, Sari, Iran, in 2010. One ml of each solution including hypotonic saline (0.3%), normal saline (0.85%), RPMI-1640 (RPMI), RPMI with 10% fetal bovine serum (FBS), RPMI with 20% FBS, ovine hydatid cyst fluid, pasteurized milk of cow, and phosphate buffered saline (PBS) along with 4×10(4) T. gondii tachyzoites were added to plate wells and incubated in 4 °C, 22 °C, 37 °C, and 37 °C under 5% CO2. The survival rate and viability assessment of parasites were performed daily and the results were analyzed using Univariate tests.

Result: Tachyzoites survival rate in PBS (4 °C) and normal saline (4 °C) were considerably high, compared to other solutions in different conditions (P<0.001). The best temperature for Toxoplasma maintenance was 4 °C (P<0.001).

Conclusion: This study introduces two available and economical solutions, PBS (4 °C) and normal saline (4 °C) media, for maintenance of Toxoplasma tachyzoites as appropriate choice media for a noticeable period of time (11 days) in vitro.

No MeSH data available.


Related in: MedlinePlus

Survival time of T. gondii tachyzoites at different temperature in the eight kinds of cell-free media
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Figure 1: Survival time of T. gondii tachyzoites at different temperature in the eight kinds of cell-free media

Mentions: The highest survival rate of tachyzoites was observed in PBS followed by normal saline (P<0.001) (Fig. 1). Additionally, the tachyzoites survival rate was varied for each medium at various temperatures. In the other media, Toxoplasma remained stable for a short period and then slightly regressed (Fig. 2). In addition, Toxoplasma survival times in these media are influenced by the temperature. The best temperature for Toxoplasma preservation was 4 °C (P<0.001). In addition, interaction between temperature degrees and different media was statistically significant (P<0.001) (Table 1). For survey of Toxoplasma infectivity, 0.5 ml of each solution containing tachyzoites was injected to one mouse peritoneum that resulted in mice death during 3–5 d.


Comparison of Eight Cell-Free Media for Maintenance of Toxoplasma gondii Tachyzoites.

Kalani H, Daryani A, Sharif M, Ahmadpour E, Alizadeh A, Nasrolahei M, Sarvi S, Kalani F, Faridnia R - Iran J Parasitol (2016 Jan-Mar)

Survival time of T. gondii tachyzoites at different temperature in the eight kinds of cell-free media
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4835459&req=5

Figure 1: Survival time of T. gondii tachyzoites at different temperature in the eight kinds of cell-free media
Mentions: The highest survival rate of tachyzoites was observed in PBS followed by normal saline (P<0.001) (Fig. 1). Additionally, the tachyzoites survival rate was varied for each medium at various temperatures. In the other media, Toxoplasma remained stable for a short period and then slightly regressed (Fig. 2). In addition, Toxoplasma survival times in these media are influenced by the temperature. The best temperature for Toxoplasma preservation was 4 °C (P<0.001). In addition, interaction between temperature degrees and different media was statistically significant (P<0.001) (Table 1). For survey of Toxoplasma infectivity, 0.5 ml of each solution containing tachyzoites was injected to one mouse peritoneum that resulted in mice death during 3–5 d.

Bottom Line: The survival rate and viability assessment of parasites were performed daily and the results were analyzed using Univariate tests.Tachyzoites survival rate in PBS (4 °C) and normal saline (4 °C) were considerably high, compared to other solutions in different conditions (P<0.001).The best temperature for Toxoplasma maintenance was 4 °C (P<0.001).

View Article: PubMed Central - PubMed

Affiliation: Toxoplasmosis Research Center, Mazandaran University of Medical Sciences, Sari, Iran.

ABSTRACT

Background: Toxoplasmosis is considered as one of the most common infectious diseases caused by the protozoan parasite Toxoplasma gondii. Tachyzoite is the main form of Toxoplasma and continuously is maintained in cell culture or injected into the mice peritoneal cavity. This study was designed to evaluate the survival rate of RH strain of T. gondii tachyzoites in different cell free, nutrient and biological media at different temperatures.

Methods: This experimental study was performed at the Toxoplasmosis Research Center, Mazandaran University of Medical Sciences, Sari, Iran, in 2010. One ml of each solution including hypotonic saline (0.3%), normal saline (0.85%), RPMI-1640 (RPMI), RPMI with 10% fetal bovine serum (FBS), RPMI with 20% FBS, ovine hydatid cyst fluid, pasteurized milk of cow, and phosphate buffered saline (PBS) along with 4×10(4) T. gondii tachyzoites were added to plate wells and incubated in 4 °C, 22 °C, 37 °C, and 37 °C under 5% CO2. The survival rate and viability assessment of parasites were performed daily and the results were analyzed using Univariate tests.

Result: Tachyzoites survival rate in PBS (4 °C) and normal saline (4 °C) were considerably high, compared to other solutions in different conditions (P<0.001). The best temperature for Toxoplasma maintenance was 4 °C (P<0.001).

Conclusion: This study introduces two available and economical solutions, PBS (4 °C) and normal saline (4 °C) media, for maintenance of Toxoplasma tachyzoites as appropriate choice media for a noticeable period of time (11 days) in vitro.

No MeSH data available.


Related in: MedlinePlus