We investigated mass mortalities of koi, Cyprinus carpio Linnaeus, 1758, experienced in South Indian fish farms by virus isolation, electron microscopy, PCR detection, sequencing of capsid protein gene and transmission studies. Samples of moribund koi brought to the laboratory suffered continuous mortality exhibiting swimming abnormalities, intermittent surfacing and skin darkening. Irido-like virus was isolated from the infected fish in the indigenous snakehead kidney cell line (SNKD2a). Icosahedral virus particles of 100 to 120 nm were observed in the infected cell cultures, budding from the cell membrane. Virus transmission and pathogenicity studies revealed that horizontal transmission occurred associated with mortality. PCR analysis of infected fish and cell cultures confirmed the presence of Ranavirus capsid protein sequences. Sequence analysis of the major capsid protein gene showed an identity of 99.9% to that of largemouth bass virus isolated from North America. Detection and successful isolation of this viral agent becomes the first record of isolation of a virus resembling Santee-Cooper Ranavirus from a koi and from India. We propose the name koi ranavirus to this agent.

We isolated the infectious viral agent from infected Asian seabass, Lates calcarifer (Bloch, 1790) using SSN‐1 cells. The isolated virus was cultivated in indigenous marine fish cell lines developed from ornamental sea anemone fish Amphiprion sebae and from Asian seabass. Reverse transcriptase PCR using betanodavirus‐specific primers gave positive results for the RNA from virus‐infected cells (hereinafter referred to as L. calcarifer nervous necrosis virus Indian strain – LCNNV‐In01). Serum neutralization test indicated partial homogeneity between Atlantic halibut nervous necrosis virus (AHNNV 692/9/98) and LCNNV‐In01. Transmission electron micrographs of the virus‐infected cell culture showed large numbers of 25–30‐nm particles in cellular vacuoles with features similar to betanodaviruses. Pathogenicity study in seabass fingerlings using cell culture grown virus revealed nervous necrosis in retinal cells following a 21‐day challenge trial. The nodavirus was reisolated from the infected fish using clownfish and seabass cell lines. Sequence analysis of the coat protein gene showed that the virus belonged to RGNNV group (GenBank Acc. No. ). The isolation of the nodavirus from seabass in India and its potential to grow in non‐homologous cell lines indicate the serious threat of the Indian strain of betanodavirus (LCNNV‐In01) to cultured and ornamental marine fish that have their hatchery production successfully standardized in India.

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Background: To date, very few small drug molecules are used for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has been discovered since the epidemic commenced in November 2019. SARS-CoV-2 RdRp and spike protein are essential targets for drug development amidst whole variants of coronaviruses. Objective: This study aims to discover and recognize the most effective and promising small molecules against SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) and spike protein targets through molecular docking screening of 39 phytochemicals from five different Ayurveda medicinal plants. Methods: The phytochemicals were downloaded from PubChem, and SARS-CoV-2 RdRp and spike protein were taken from the protein data bank. The molecular interactions, binding energy, and ADMET properties were analyzed. Results: Molecular docking analysis identified some phytochemicals, oleanolic acid, friedelin, serratagenic acid, uncinatone, clemaphnol A, sennosides B, trilobine and isotrilobine from ayurvedic medicinal plants possessing greater affinity against SARS-CoV-2-RdRp and spike protein targets. Two molecules, namely oleanolic acid and sennosides B, with low binding energies, were the most promising. Furthermore, based on the docking score, we carried out MD simulations for the oleanolic acid and sennosides B-protein complexes. Conclusion: Molecular ADMET profile estimation showed that the docked phytochemicals were safe. The present study suggested that active phytochemicals from medicinal plants could inhibit RdRp and spike protein of SARS-CoV-2.