Susceptibility of orange chromide, Etroplus maculatus (Bloch, 1795) to experimental infection of Betanodavirus

Abstract

Viral nervous necrosis (VNN), or viral encephalopathy and retinopathy caused by nervous necrosis virus (NNV), is an acute viral disease of fish, resulting in high mortality globally in cultured marine fish. The pathogen, Betanodavirus belonging to the family Nodaviridae, has been widely documented in both farmed and wild fish, besides in invertebrate species across the habitat barriers. However, cichlid fish, in particular Etroplus species, have a low susceptibility or resistance to this disease. The present study was undertaken to check the susceptibility of a brackishwater cichlid, orange chromide (Etroplus maculatus) through intramuscular injection and immersion challenge by NNV (RGNNV genotype). Intramuscular injection (10 μl) with NNV (105 viral particles ml−1) resulted in cumulative mortality of 50% at 15 days of post-infection (dpi), while immersion challenge in NNV spiked sea water (105 TCID50 ml−1) resulting in a mortality of 25% at 19 dpi. Unlike many marine fish which showed 100% mortality within 3–5 days, E. maculatus challenged through both the routes exhibited severe clinical symptoms for 2–3 weeks followed by a recovery stage and moderate level of mortality. Reverse transcription polymerase chain reaction (RT-PCR) tests on live fish sampled during the course of the experiment and the dead fish exhibiting symptoms in both challenge groups tested NNV-positive. Histological examination of fish in challenged groups showed VNN-specific lesions characterized by vacuolations in the brain and eye with more severe lesions in the injected group than the immersion group. In survived fish, no clinical sign was observed, but NNV could be detected by RT-PCR with moderate to severe vacuolations in the brain and eyes until 30 dpi. Despite the virus entering the body and triggering typical symptoms of VNN, the fish appeared to exhibit moderate resistance with a survival of 50–75%. The results indicate that E. maculatus could serve as a brackishwater fish model to study host-pathogen interaction and development of resistance for potential application in NNV-vaccine studies.