First report on the occurrence of cyprinid herpesvirus 3 in koi carp (Cyprinus carpio koi) in India.

This study reports the occurrence of cyprinid herpesvirus 3 (CyHV-3) in koi carp (Cyprinus carpio koi) for the first time in India. The koi carp, with clinical signs of ulcer with haemorrhage on body surface, necrosis of fin and discolouration of gill associated with huge mortality, were observed in aquarium shops, rearing tanks and grow-out ponds located in Chennai, India. The PCR assay carried out on infected fish samples using different primer sets specific to CyHV-3 confirmed its presence in the infected fish. Sequence analysis of partial thymidine kinase gene revealed 100% similarity with the sequence of CyHV-3 available in GenBank. Cell lines of koi carp and catla were found to be susceptible to CyHV-3 and its replication was confirmed by viral-specific cytopathic effect, PCR and bioassay. The CyHV-3 infection was reproduced by intramuscular injection of inoculum prepared from CyHV-3-infected fish to satisfy Koch's postulates. Tissue tropism of CyHV-3 in infected fish by PCR assay revealed the presence of CyHV-3 in all vital organs with prominent band in gill and gut tissue.

First report on the occurrence of white spot syndrome virus, infectious myonecrosis virus and Enterocytozoon hepatopenaei in Penaeus vannamei reared in freshwater systems.

Samples of white leg shrimp, Penaeus vannamei, were collected on a monthly basis from freshwater ponds with the salinity of 0 ppt located at Tiruvannamalai and Villupuram districts in Tamil Nadu, India for screening of viral and fungal pathogens. Totally, 130 shrimp samples were collected from 67 freshwater ponds and screened for white spot syndrome virus (WSSV), infectious myonecrosis virus (IMNV), infectious hypodermal and haematopoietic necrosis virus (IHHNV) and Enterocytozoon hepatopenaei (EHP) by PCR and RT-PCR using pathogen-specific primers. Among the samples screened, one sample was found to be positive to WSSV, two samples showed positive to IMNV and two samples positive for EHP. No sample showed positive to IHHNV. The WSSV detected in the sample was found to be a new strain of WSSV and highly virulent. The inoculum prepared from freshwater reared WSSV or IMNV-infected shrimp caused 100% mortality in experimental infection studies. The PCR and RT-PCR results revealed the presence of WSSV and IMNV in different organs of experimentally infected shrimp, respectively. No clinical signs were observed in experimentally EHP-injected shrimp, although the PCR results revealed the presence of EHP in experimentally infected shrimp.

In vitro propagation of tilapia lake virus in cell lines developed from Oreochromis mossambicus.

Tilapia lake virus (TiLV)-suspected samples of tilapia were collected from grow-out ponds located with clinical signs and mortality ranged from 5% to 50%. The reverse transcription-polymerase chain reaction (RT-PCR) assay revealed the presence of TiLV in the disease outbreak ponds. Cell lines were developed from heart, gill and eye of Mozambique tilapia and characterized. Morphologically, these cell lines are composed of epithelioid cells. The optimum growth of these cells was observed at 28°C and 20% concentration of FBS. After cryopreservation, 70%-90% of cells were found to be viable. The cells of all three cell lines were found to be positive to fibronectin and pancytokeratin. PCR amplification of 16S rRNA and COI of O. mossambicus confirmed the origin of these cell lines from O. mossambicus. Heart and gill cell lines were found to be highly susceptible to TiLV and found to be useful for its isolation from infected fish samples. The experimental infection was carried out in O. niloticus and O. mossambicus using the TiLV propagated in susceptible cell lines. The RT-PCR results revealed the presence of TiLV in brain, gill, liver, kidney, spleen, eye, muscle, intestine and heart of experimentally infected O. niloticus and O. mossambicus.