Serum neutralising antibody titres against a lineage 2 neuroinvasive West Nile Virus strain in response to vaccination with an inactivated lineage 1 vaccine in a European endemic area.

In the last decade in Hungary and the neighbouring countries, West Nile Neuroinvasive Disease (WNND) has been caused in dramatically increasing numbers by lineage 2 West Nile Virus (WNV) strains both in horses and in humans. The disease in this geographical region is seasonal, so vaccination of horses should be carefully scheduled to maintain the highest antibody titres during outbreak periods. The objective of this study was to characterise the serum neutralising (SN) antibody titres against a lineage 2 WNV strain in response to vaccination with an inactivated lineage 1 vaccine (Equip® WNV). Thirty-two seronegative horses were enrolled in the study, 22 horses were allocated to the vaccinated group and 10 retained as unvaccinated controls. Horses were vaccinated according to the product's vaccination guidelines. A primary vaccination of two doses administered 28 days apart was initiated approximately 5 months before the WNV outbreak season, followed by a booster vaccination one year later. Blood samples were collected during a 2-year period to monitor production of SN antibodies against lineage 1 and the enzootic lineage 2 WNV strain. Mean antibody titres against lineage 1 WNV were significantly higher (P ≤ 0.05) in the vaccinated group compared to the control group at all-time points after the primary dose of vaccination. Similarly, mean antibody titres against lineage 2 WNV were significantly higher (P ≤ 0.05) in the vaccinated group compared to the control group at all time-points except at 6 months after the primary vaccination. SN antibody titres were significantly higher against lineage 1 than lineage 2 at all-time points. According to the results, vaccination with an inactivated lineage 1 vaccine induces antibodies against both WNV lineages 1 and 2 strains up to 2 years after booster vaccination, but in those geographical regions where lineage 2 strains are responsible for seasonal outbreaks, a booster vaccination should be considered earlier than 12 months after primary vaccination.

West Nile virus host-vector-pathogen interactions in a colonial raptor.

BACKGROUND: Avian host species have different roles in the amplification and maintenance of West Nile virus (WNV), therefore identifying key taxa is vital in understanding WNV epidemics. Here, we present a comprehensive case study conducted on red-footed falcons, where host-vector, vector-virus and host-virus interactions were simultaneously studied to evaluate host species contribution to WNV circulation qualitatively. RESULTS: Mosquitoes were trapped inside red-footed falcon nest-boxes by a method originally developed for the capture of blackflies and midges. We showed that this approach is also efficient for trapping mosquitoes and that the number of trapped vectors is a function of host attraction. Brood size and nestling age had a positive effect on the number of attracted Culex pipiens individuals while the blood-feeding success rate of both dominant Culex species (Culex pipiens and Culex modestus) markedly decreased after the nestlings reached 14 days of age. Using RT-PCR, we showed that WNV was present in these mosquitoes with 4.2% (CI: 0.9-7.5%) prevalence. We did not detect WNV in any of the nestling blood samples. However, a relatively high seroprevalence (25.4% CI: 18.8-33.2%) was detected with an enzyme-linked immunoabsorbent assay (ELISA). Using the ELISA OD ratios as a proxy to antibody titers, we showed that older seropositive nestlings have lower antibody levels than their younger conspecifics and that hatching order negatively influences antibody levels in broods with seropositive nestlings. CONCLUSIONS: Red-footed falcons in the studied system are exposed to a local sylvatic WNV circulation, and the risk of infection is higher for younger nestlings. However, the lack of individuals with viremia and the high WNV seroprevalence, indicate that either host has a very short viremic period or that a large percentage of nestlings in the population receive maternal antibodies. This latter assumption is supported by the age and hatching order dependence of antibody levels found for seropositive nestlings. Considering the temporal pattern in mosquito feeding success, maternal immunity may be effective in protecting progeny against WNV infection despite the short antibody half-life measured in various other species. We conclude that red-footed falcons seem to have low WNV host competence and are unlikely to be effective virus reservoirs in the studied region.

Comparison of assays for the detection of West Nile virus antibodies in equine serum after natural infection or vaccination.

West Nile virus (WNV) mainly infects birds, horses and humans. Outcomes of the infection range from mild uncharacteristic signs to fatal neurologic disease. The main objectives of the present study were to measure serum IgG and IgM antibodies in naturally exposed and vaccinated horses and to compare results of haemagglutination inhibition test (HIT), enzyme-linked immunosorbent assay (ELISA) and plaque reduction neutralisation test (PRNT). Altogether 224 animals were tested by HIT for WNV antibodies and 41 horses were simultaneously examined by ELISA and PRNT. After primary screening for WNV antibodies, horses were vaccinated. Samples were taken immediately before and 3-5 weeks after each vaccination. McNemar's chi-squared and percent agreement tests were used to detect concordance between HIT, ELISA and PRNT. Analyses by HIT confirmed the presence of WNV antibodies in 27/105 (26%) naturally exposed horses. Sera from 57/66 (86%) vaccinated animals were positive before the first booster and from 11/11 (100%) before the second booster. HIT was less sensitive for detecting IgG antibodies. We could detect postvaccination IgM in 13 cases with IgM antibody capture ELISA (MAC-ELISA) and in 7 cases with HIT. WNV is endemic in Hungary and regularly causes natural infections. Protective antibodies could not be measured in some of the cases 12 months after primary vaccinations; protection is more reliable after the first yearly booster. Based on our findings it was not possible to differentiate infected from recently vaccinated horses using MAC-ELISA. HIT cannot be used as a substitute for ELISA or PRNT when detecting IgG, but it proved to be a useful tool in this study to gain statistical information about the tendencies within a fixed population of horses.