RESUMEN
The Maedi-visna virus (MVV) causes a persistent infection in small ruminants, and its high genetic heterogeneity affects the performance of diagnostic tests when used in different populations. Therefore, the aim of this study was to develop a bead-based multiplex immunoassay tailored to detect antibodies against a Norwegian MVV strain. We used tissue samples from 14 PCR-positive sheep from a recent MVV outbreak in Norway to sequence the viral strain and produced recombinant antigens based on sequences from one animal. The assay included commercial TM-A and recombinant Norwegian p25, p16-25 and SU5 antigens. Cut-off values for each antigen were determined using receiver operating characteristic curves on 40 ELISA-negative and 67 ELISA-positive samples from the outbreak. The intraplate and interplate repeatability were investigated by testing a quadruplicate of five samples over three days, while the analytical sensitivity (aSe) and specificity (aSp) were measured in comparison to a commercial ELISA. The repeatability showed a coefficient of variation below 15% for most positive samples. The aSe was equal or higher for the multiplex assay than the ELISA, and the aSp of each antigen was 91.7, 93.3, 95.0 and 93.3% for p25, p16-25, SU5 and TM-A, respectively. The assay shows promising results; however, further evaluations of diagnostic characteristics are necessary before implementation in the Norwegian surveillance programme.
RESUMEN
Lake Steinsfjorden, an important Norwegian location for noble crayfish (Astacus astacus), is often affected by cyanobacterial blooms caused by microcystin (MC)-producing Planktothrix spp. The impact of MCs on noble crayfish as a food source and crayfish health is largely unknown. We investigated the quantities and correlations of MCs in noble crayfish and lake water during and after a cyanobacterial bloom peaking in June-July 2015. Noble crayfish and water samples were collected monthly from June to October 2015 and in October 2016. The content of MCs was analysed by ELISA from tail muscle, intestine, stomach and hepatopancreas. PCR analysis for Planktothrix gene markers was performed on crayfish stomach content. Water samples were analysed for phytoplankton composition, biomass and MCs. PCR-positive stomach contents indicated Planktothrix to be part of the noble crayfish diet. Concentrations of MCs were highest in the hepatopancreas, stomach and intestine, peaking in August-September. Tail muscle contained low concentrations of MCs. Similar levels of MCs were found in crayfish from 2016. Except in September 2015, a normal portion of boiled noble crayfish tails was below the tolerable daily intake (TDI) for MCs for humans. Removing the intestine more than halved the content of MCs and seems a reasonable precautionary measure for noble crayfish consumers.
