Consumption of unpasteurized cow milk is can be a source of infection in human. Reports shown the presence of T. gondiiin milk of lactating mammals such as sheep and camel. Grazing animals may come into contact with T. gondii oocyst in the grass passed by stray cats and later passed to the infant through lactating. Thus the aim of this study is to study the presence of T.gondii DNA in cow’s fresh milk. 15ml of milk was collected and stored at -40°C. 300µL of milk was used for DNA extraction using Genomic Mini DNA Kit. PCR was done using primer 5’-AAGCTTATGCGAGGCGGGACG-3’ and 5’-GATATCTCACTGCTTAATTTT CTCACACGTCACGG-3’. The reaction consisted of 31 cycles with the following conditions: 30 second at 98°C (denaturation), followed by 31 cycles at 98°C for 30 second (denaturation), 5 second at 64°C (annealing), and 30 second at 72°C (extension), final extension step of 10 minutes at 72°C and stopped at 16°C. Electrophoresis was done on 1% agarose gel stained with ethidium bromide (2µl). Mice peritoneal fluid infected with T.gondiiwas used as the positive control. The expected PCR product size (MIC3 gene form T.gondii) was 1080 base pairs. Results from the gel electrophoresis showed no band was formed for the milk samples. The fresh milk from Klang, UPM and Shah Alam were free from T. gondiihowever further studies should be conducted to detect other microorganisms that may be present in the milk to assure safe consumption.
The intramuscular injection in mice always lead to a great difficulty because of its small size. This study was designed to compare the effectiveness of intramuscular routes and intraperitoneal routes in the delivery of pcDNA3.1/HisB-MIC3 as a DNA vaccine towards toxoplasmosis. In this study two set of mice (A and B), each set were divided into four groups. Group one was injected with sterile distilled water, group two was injected with 50 µg of empty pcDNA3.1/His B, group three was injected with 50 µg of pcDNA3.1/His B-MIC3 and group four was not injected at all. The mice in set A was injected through intramuscular route and mice in set B was injected through intraperitoneal route. The IgG titre was determined using Mouse Toxoplasma IgG ELISA Kit. Both group injected with pcDNA3.1/His B-MIC3 as vaccine showed increase in IgG titre. Mice injected with pcDNA3.1/His B-MIC3 in intramuscularly shows about the same IgG titre with mean (0.128 ± 0.005 IU/ml) compared to mice injected with the same vaccine through intraperitoneal injection with IgG titre mean (0.11 ±0.021IU/ml). This result suggested that the is no significant different in IgG titre in mice injected through intramuscular injection or intraperitoneal injection. So, in the future intraperitoneal injection can be used to study the efficacy of the vaccine in mice.