Detection of scale drop disease virus from non-destructive samples and ectoparasites of Asian sea bass, Lates calcarifer.

Non-destructive sampling methods offer practical advantages to detection and monitoring of viral pathogens in economically important farmed fish and broodstock. Here, we investigated whether blood, mucus and fin can be used as non-lethal sample sources for detection of scale drop disease virus (SDDV) in farmed Asian sea bass, Lates calcarifer. Detection of SDDV was performed in parallel from three non-destructive and seven destructive sample types, collected from both clinically sick fish and subclinical fish obtained from an affected farm. The results showed that SDDV was detectable in all 10 sample types with the percentage ranging from 20% to 100%. Blood was the best non-destructive sample source exhibited by the fact that it yielded 100% SDDV-positive tests from both sick (n = 12, 95% CI: 69.9-99.2) and clinically healthy fish (n = 4, 95% CI: 39.6%-97.4%) and is considered a "sterile" sample. This study also revealed concurrent infection of SDDV and two ectoparasites Lernanthropus sp. and Diplectanum sp., in all affected fish (n = 8, 95% CI: 46.7-99.3) during the disease outbreak. These ectoparasites also tested positive for SDDV by PCR, indicating that they were potential sample sources for PCR-based detection of SDDV and possibly other viruses infecting Asian sea bass.

A validated semi-nested PCR for rapid detection of scale drop disease virus (SDDV) in Asian sea bass (Lates calcarifer).

Scale drop diseases virus (SDDV), a newly characterized virus of farmed Asian sea bass (Lates calcarifer), has been reported in several countries in Southeast Asia. However, no fully validated detection method is publicly available for disease diagnosis and surveillance. Here, we described a newly developed semi-nested PCR (snPCR) method for detection of the virus from field samples. The designed primers targeting a gene encoding ATPase generated amplicons of 738 bp and 412 bp in the first and second step PCR, respectively. The established protocol could detect down to 100 viral copies/µL template and was 100-fold more sensitive than single step PCR. A Specificity test against extracted DNA from ten bacterial pathogens, tissues from viral infected specimens and fish host revealed no cross amplification. The SDDV snPCR method could detect the virus from all clinical samples showing symptoms of scale drop disease (n = 25) and all samples from outbreaks of an unknown disease (n = 6) whereas all clinically healthy fish sea bass (n = 161) and grouper (n = 45) collected from different provinces tested negative. The newly established protocol might be useful for Asian sea bass farming countries to initiate disease diagnosis and surveillance.