Changes in viral load in different organs of Japanese Encephalitis virus-infected chick embryo under the influence of Belladonna 200C

Abstract

Background: Japanese encephalitis(JE) is highly prevalent in many states of India. Belladonna 200C is widely used in the prevention and treatment of JE.The effect of Belladonna 200C in virus replication inside different tissues has not been studied. Objective: To study the effect of Belladonna 200C in virus replication inside different tissues utilising chick embryo model. Materials and Methods: Twelve-day-old fertilised eggs of Black Australorp were inoculated with JE via chorioallantoic membrane (CAM) route in different experimental sets: infection, Belladonna 200C treated and vehicle control, keeping matched blank sets. All experimental sets were incubated for 48 hours. After incubation, viable eggs were sacrificed humanly and different tissues were observed and collected for viral load determination by real-time-polymerase chain reaction (PCR). Results: The control group showed visible pocks over the CAM; brains were liquefied due to haemorrhagic liquefactive necrosis and white patches were found over the liver. However, the medicine-treated group was apparently normal; there were no visible changes in the brain and the liver was healthy like control. Real-time-PCR results showed high viral load in CAM and brain with absence of viral RNA in liver of the virus-infected group. Pre-treatment with Belladonna 200C significantly reduced the overall load (P < 0.05) in CAM and brain which correlated with the morbid pathological changes of the organs. Conclusion: Although Belladonna 200C did not completely inhibit JE viral replication in the brain, it reduced the severity of JE by diminishing the viral loads in this tissue.