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Email the author (Login required) Implementation of the mixed linear models in experimental infections with WSSV in the withe shrimp Litopenaeus vannamei
DOI: https://doi.org/10.3856/vol43-issue3-fulltext-17
Abstract
The White Spot Syndrome Virus has been very detrimental for the shrimp industry. Up to date there is no cure for the disease, thus, it is necessary to implement reliable experimental strategies to evaluate the effect of drugs and the host response during the prognosis of the disease. In this study, we evaluated two ways of infection with WSSV (200 copies WSSV-DNA) (injection and immersion), at constant temperature (26 ± 0.5ºC), in juveniles of Litopenaeus vannamei (4.8 ± 0.38 g) in intermolt stage. In the infection by injection, the organisms were lethargic with reddish appearance 2 days after infection and mortality (100%) was observed within 2-5 days: 63% organisms with light degree of infection [20 copies of DNA & 1-5 Cowdry A-type inclusions in hypertrophied nuclei (CAI)/200 fields], 21% had moderate infection (200 copies of DNA & 1-2 CAI/20 fields) and 16% severely infected (2000 copies of DNA & more than 10 CAI/field). No mortality was observed in the controls. In the infection by immersion, the signs of WSSV were observed 3 days after infection: 38% of mortality was observed during 3-9 days: 25% of the organism with light degree of infection (20 copies of DNA & 1-2 CAI/20 fields), 5% moderately infected (200 copies of DNA & 1-2 CAI/20 fields) and 8% with severe infection (2000 copies of DNA & 1-5 CAI/2 fields. The other 62% organisms were necropsied at day 12 and tested positive to the WSSV-PCR (light infection = 20 copies of DNA), but did not show CAI by histology. No mortality was observed in the controls. For the statistical analysis, the data did not fit the criteria of independence and linearity needed for the analyses of variance, thus we used instead the mixed linear models and were able to observe a better prediction: in the injected organisms, the mortality reached the highest peak at day 5 after infection. In the organisms infected by immersion, the highest peak or mortality was observed at day 9 after infection. The analysis of variance of Kenward-Roger indicated significant differences between the days of mortality (F = 20.1, P = 0.001), as well as among the ways of infection (Random analysis) (P = 0.007).
