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.