Figure 3

Prevalence of anti-RHDV and anti-RHDV2 antibodies in a wild rabbit population analyzed by indirect ELISA. A A collection of 180 sera from wild rabbits captured in a region of Central Spain (Ciudad Real) during the months of April and November 2014 was tested by ELISA using plates coated with RHDV or RHDV2 VLPs. A single dilution (1/500) of each sample was tested and recorded OD values for each sample are represented as two separate groups. A line at an arbitrary value (0.155) corresponding to the mean OD+ 2 times the standard deviation of 10 samples from non vaccinated animals at the same dilution is shown. Mean plus SD are indicated for each antigen group. Asterisks indicate a statistically significant difference (p < 0.0001) as assessed by a paired t-test. B Representation of OD values obtained using three different dilutions (1/300, 1/900 and 1/2700) of 15 selected sera from the same collection. ELISAs were performed in duplicate under the same conditions indicated above. Arrows highlight sera samples showing reactivity with RHDV2 but not with RHDV VLPs. Procedures involving obtainment of animal samples were carried out under EU guidelines and supervised and approved by the institutional ethical review committee.