Effects of ethanolic extract of neem leaves on the sporocyst of Sarcocystis availability in vitro

The aim of this study was to evaluate the in vitro anti-sporocyst efficiency of Azadirachta indica (neem) leaves ethanolic extracts against Sarcocystis and using it as alternative therapy for Sarcocystis in dogs. 100 g of faeces collected from experimentally-infected puppies with Sarcocystis by feeding them on 300 g meat (oesophagus and diaphragm) of sheep infected with Sarcocystis. The faeces melted into distilled water and then the sample was centrifuged. After dividing the precipitate into sixteen equal samples, a sample was randomly selected and inspected using the flotation (sugar solution) technique, then the quantity of sporocysts was calculated using a haemocytometer, which was 5000 individuals/g. Afterward, the samples were divided to 3 group each containing on five samples. Finally, 3 mL from the neem extract was added in different concentrations (250, 500, 750 and 1000 mg; C1, C2, C3 and C4 respectively) and placed in incubator at 37 ºC. The first group was inspected after 24 h, the second after 48 h, and the third after 72 h. A significant reduction was observed in the number of sporocysts in the samples treated with high concentration of neem leaves extract. The number of sporocysts become zero in the sample with the higher dose (C4; 1000 mg mL-1) at 48 h and 72 h of incubation, and also significant reduction in number was observed at 24 h. The number of sporocysts was nil after 72 h of incubation in C3 (750 mg mL-1), and much lower after 48 and 24 h. In C2 (500 mg mL-1), the quantity of sporocysts was zero after 72 h of incubation and decreased after 48 h. The minimal impact was observed in C1 (250 mg mL-1). The best antisporocyst action was observed at C4 (1000 mg mL-1). As a result, it is suggested that an ethanolic extract of neem leaf at 1000 mg mL-1 can be utilized in the in vitro therapy of Sarcocystis.