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In vitro evaluation of antiviral activity of leaf extracts of Azadirachtaÿindica, Moringaÿoleifera, and Morusÿalba against the foot and mouth disease virus on BHK-21 cell line

IshratÿImran^a, ImranÿAltaf^b, MuhammadÿAshraf^a, AqeelÿJaveed^a, NeelmaÿMunir^c, RasheedaÿBashir^c,*

 
ABSTRACT:     Plants are widely used for the treatment of different ailments including viral infections. There is a great need to explore antiviral agents of herbal origin to combat the viral resistance. Foot and mouth disease (FMD) is highly infectious viral disease of cloven-footed animals. Three local Pakistani plants Azadirachta indica (AI), Moringa oleifera (MO) and Morus alba (MA) are reported to possess antiviral activity against DNA and RNA viruses. In the present study ethanolic leaf extracts of these plants were evaluated for their cytotoxicity against BHK-21 (baby hamster kidney) cell line and antiviral activity against FMD virus by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) colourimetric assay. Ethanolic extracts of each plant were obtained and stock solution of 40ÿmg/ml was prepared in cell culture media (M-199) and sterilized by filtration. Two-fold serial dilutions of 200ÿµg/ml, 100ÿµg/ml, 50ÿµg/ml, 25ÿµg/ml, 12ÿµg/ml, 6ÿµg/ml, and 1ÿµg/ml of each plant were prepared in M-199 cell culture media. The BHK-21 cell line was identified, purified, and characterized. Tissue culture infective dose (TCID₅₀) for FMD virus was calculated by ten-fold dilutions of FMD virus. The virus was used at the rate of 10⁶ TCID₅₀. Ethanolic extract of AI exhibited stronger anti-FMD virus activity than MO and MA at a concentration nontoxic to the BHK-21 cell line. These plants may be helpful to elucidate effective antiviral agents against FMD virus.

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* Corresponding author, E-mail: rashidasbs@yahoo.com

Received 27 May 2017, Accepted 0 0000