In Ovo Antiviral Screening of Cholistani Plants Against Swine Influenza Virus and Confirmation Through Real Time PCR

Abstract

Cholistani plants are a rich source of many biological/pharmacological agents. Recent studies have supported the idea that cholistani plants are rich in antimicrobial agents especially against viruses. Medicinal plants like Haloxylon recurvum, Salsola baryosma, and Achyranthes aspera hold the potential to control viruses like Avian Influenza and Swine Influenza. Prevention by vaccination is the best treatment so far, but some side effects are associated with vaccines alone or in combination with other influenza virus strains. Eleven different Cholistani plants (H. recurvum, H. salicornicum, A. aspera, Suaeda fruticosa, Sporobolus icolados, Solanum surattense, Ochthochloa compressa, Neurada procumbens, Panicum antidotal, Oxystelma esculentum, and Salsola baryosma) were selected and tested against SI strian H1N1. The samples were washed and dried under shade at RT. All the plants were ground and used in making extracts using different solvents. The SIV H1N1 isolate was confirmed through real-time (rt) PCR and then subjected to 9-11 days old chick embryonated eggs. Eggs were candled and viability of embroy was confirmed. After 48hrs, allantoic fluids were collected, and viral loads were also confirmed through HA test and RT-PCR. Most of the extracts were found effective in controlling growth of the virus. Among these, n-hexane and ethanol (EtOH) extracts were found most effective, and all extracts were active against SIV H1N1. In the case of ethyl acetate and methanol (MtOH) 9 out of 11, similarly, in n-butanol (BtOH),7 out of 11 extracts were active against SIV H1N1. All the results were first confirmed HA test and later through rt-PCR. The virus was not detected in the outcomes of rt-PCR, confirming the antiviral potentials of extracts of Cholistani plants. This study strongly supported the idea that Cholistani plants are a rich source of antiviral agents and can be used in pure and crude form.