The antiviral activities of Opuntia delinii, Opuntia manocantha, and Opuntia stricta were evaluated against Peste des petits ruminants virus (PPRV) in this study, as these plants are associated with a lot of antiviral activity as shown by literature review. Ethanolic and aqueous extracts of all the three species of Opuntia were obtained by using soxhlet apparatus (Davey et al. 2010) but first crushed them into small pieces with a sharp knife to have better extraction results. The resultant extracts were dried in rotary evaporator using standard operating procedures until semisolid extract was obtained. Different dilutions were made by dissolving in double distilled water. Vero cells were made mildly affected by mild strains of Peste des petits ruminants virus. Dilutions of these extracts were applied on Vero cell line in triplicate manner that was first made confluent up to 90% in 96 well cell culture plates. For performing anti viral assay, Positive control and negative controls used were media plus cells and virus plus media respectively. These plates were incubated for a period of four days. After this incubation period, viability of cells was determined by MTT colorimetric assay i.e. number of living and dead cells. The cytotoxic activity of above mentioned three plant species was performed by treating the Vero cells with different dilutions as used in antiviral assay and incubating the 96 well plates for 4 days. Viability of cells was determined by MTT assay. The positive and negative control for cytotoxic evaluation was cells plus media and cells plus media plus DMSO (Dimethyl sulfoxide) 5% respectively. Results were calculated in terms of cell survival percentage (CSP) for anti viral
and death rate (%) for cytotoxic assay. At highest concentrations, i.e.500 to 1000 µg/ml, all the ethanolic and aqueous extracts obtained from all the plant species showed cytotoxicity but at the lower concentrations ranging from 7.81µg/ml to 125µg/ml, there was no cytotoxicity. Antiviral and cytotoxic activity of the plant extracts was evaluated by applying Analysis Of Variance (ANOVA) and comparison between two extracts was performed by applying T-Test for statistical analysis. Statistically when these results were interpreted, they were insignificant because P value is more than 0.05. This research project has a lot of positive outcomes and future prospects. The extract of plants having good antiviral activity and with no cytotoxic activity will be good baseline for further evaluation.

CHAPTER 6
SUMMARY

The antiviral activities of Opuntia delinii, Opuntia manocantha, and Opuntia stricta were evaluated against Peste des petits ruminants virus (PPRV) in this study, as these plants are associated with a lot of antiviral activity as shown by literature review. Ethanolic and aqueous extracts of all the three species of Opuntia were obtained by using soxhlet apparatus (Davey et al. 2010) but first crushed them into small pieces with a sharp knife to have better extraction results. The resultant extracts were dried in rotary evaporator using standard operating procedures until semisolid extract was obtained. Different dilutions were made by dissolving in double distilled water. Vero cells were made mildly affected by mild strains of Peste des petits ruminants virus. Dilutions of these extracts were applied on Vero cell line in triplicate manner that was first made confluent up to 90% in 96 well cell culture plates. For performing anti viral assay, Positive control and negative controls used were media plus cells and virus plus media respectively. These plates were incubated for a period of four days. After this incubation period, viability of cells was determined by MTT colorimetric assay i.e. number of living and dead cells. The cytotoxic activity of above mentioned three plant species was performed by treating the Vero cells with different dilutions as used in antiviral assay and incubating the 96 well plates for 4 days. Viability of cells was determined by MTT assay. The positive and negative control for cytotoxic evaluation was cells plus media and cells plus media plus DMSO (Dimethyl sulfoxide) 5% respectively. Results were calculated in terms of cell survival percentage (CSP) for anti viral
and death rate (%) for cytotoxic assay. At highest concentrations, i.e.500 to 1000 µg/ml, all the ethanolic and aqueous extracts obtained from all the plant species showed cytotoxicity but at the lower concentrations ranging from 7.81µg/ml to 125µg/ml, there was no cytotoxicity. Antiviral and cytotoxic activity of the plant extracts was evaluated by applying Analysis Of Variance (ANOVA) and comparison between two extracts was performed by applying T-Test for statistical analysis. Statistically when these results were interpreted, they were insignificant because P value is more than 0.05. This research project has a lot of positive outcomes and future prospects. The extract of plants having good antiviral activity and with no cytotoxic activity will be good baseline for further evaluation.