Effect of 7DH biotherapic of Toxoplasma gondii in mice infected with the protozoan

Abstract

Introduction: Toxoplasmosis is a zoonosis caused by Toxoplasma gondii worldwide distributed [1]. In both, men and animals, the infection with T. gondii can lead to important pathologies [2]. The study of alternative treatments is important to set new therapeutic protocols, especially for the prevention of congenital toxoplasmosis. Aim: This study evaluated the effect of a biotherapic 7DH T. gondii in mice infected with T. gondii. Material and methods: The study was approved by the Ethics Committee for Animal Experimentation of the Universidade Estadual de Maringá - Protocol n° 036/2009. Fourteen mice were used - swiss male aged 57 days divided into two groups according to the treatment (or its diluent biotherapic): BIOT-200DH and Control (cereal alcohol-7%).The biotherapic was prepared with homogenized mouse brain (20 cysts of T.gondii/100μLaverage 242 bradyzoites / cyst)), according to the Brazilian Homeopathic Pharmacopoeia [3] in laminar flow. The experiment was performed as a blind randomized controlled trial. The animals were treated for 3 days immediately prior to infection. The oral treatment schedule was of 0.1mL/4x/ day, on the first day, followed by 2x/day. Animals aged 57 - 59 days were treated with biotherapic and were clinically evaluated. The animals were orally infected at the age of 60 days (20 cysts ME49-T. gondii). Within18-21 days of infection the clinical parameters were evaluated. On the 55th day of infection the eye fundus was examined (Ophthalmoscope Welch Allyn®) and the intraocular pressure was measured (Tonometer TONO-PEN® XL). After 60 days of post-infection the animals were killed in a chamber saturated with halothane, the brains were homogenized and resuspended in 1 ml of saline solution. The cysts were counted according to a rate of 25 mL of suspension, covered with 24x24 mm glass, examined in its full length. Results and discussion: The table 1 summarizes the clinical data. There was no significant difference among the groups for clinical parameters during treatment, although it was recorded the death of an animal in the biotherapic 7DH group. The dead animal presented distended stomach and liquid feces in the intestine. After the infection it was observed reduction of water consumption (p <0.01) and a reduction in the amount of feces (p = 0.052) in the group treated with biotherapic 7DHcompared to the control group. Mice treated with biotherapic 7DH developed ascites on the sixth day of post-infection, exacerbating the clinical symptoms. The number of brain cysts was compared among groups using Mann-Whitney test with 5% of significance. Although there was no significant difference among the groups (p =0.2943), the results are interesting: the number of cysts - average ± standard deviation –was of 26.7 ± 38.6 in group 7DH, and of 9.7 ± 12 , 8 in the control group. The high values for the standard deviations suggests large individual variation and the necessity of a higher number of samples for further analysis.Regarding intraocular pressure, there was a reduction (p <0.05) in the group 7DH (5.0 ± 1.0mmHg) when compared with the infection control (8.9 ± 3.8) mmHg. Figure 2 summarizes the data for fundoscopy. Once more, the sample size makes it difficult to demonstrate statistical significance, although the biological effect is clear. Conclusion: Mice pre-infection treated with biotherapic 7DH, presented bigger clinical alterations, which were measured visually and statistically compared to the control group. There was a biological effect of the biotherapic, with an increase in the number of cysts compared to the control group, without statistical significance. The group 7DH showed a significant reduction of intraocular pressure and fundoscopic analysis showed a larger number of animals without ocular changes, without statistical significance. The sample size should be reevaluated for better data interpretation and decision on the effects of the biotherapic 7DH T. gondii.