Detection of Pig Farms with Ornithodoros Erraticus by Pig Serology. Elimination of Non-specific Reactions by Carbohydrate Epitopes of Salivary Antigens

Ornithodoros erraticus is the European vector of African Swine Fever. It is therefore essential to know on which pig farms the tick is present in order to prevent contact with swine. Currently, studies are being made to ascertain this through the detection of anti-O. erraticus antibodies in the sera of swine, using three extracts from the salivary glands of the parasite (SGE): a complete extract (SGE-1), a soluble antigens extract (SGE-2), and a tissue antigens extract (SGE-3). The results of the present work show that SGE-2 gives the best differentiation between swine bitten by O. erraticus and unbitten swine in the enzyme-linked immunosorbent assay (ELISA). Using this extract, an optical density (OD) five-fold higher than the basal OD indicates that the pigs carry anti-O. erraticus antibodies. A serological study carried out in Salamanca with 8083 sera from 1756 pig farms revealed the presence of the parasite on 135 farms. However, during this study we noticed that some sera of unbitten animals gave false-positive reactions. Western blot analysis of SGE-2 of these false-positive sera demonstrated the same bands (except for two) as the real anti-O. erraticus sera. We observed, in ELISA and Western blot analysis, that such false-positive sera only recognised carbohydrate epitopes on SGE-2. This reactivity disappeared on deglycosylated SGE-2 (SGE-2-P). Therefore, SGE-2-P is the antigen that confers the greatest specificity to serology. In this study it was also observed that the low levels of anti-O. erraticus antibodies found in some cases may be because the swine were bitten some months previously on a different farm or that the current farm harboured only a few specimens of O. erraticus, so pig-tick contact is unlikely and hence the pigs either only develop a primary response or the time between contacts is very long and the levels of antibodies fall. Since pigs could be bitten on a different farm, the presence of low levels of anti-O. erraticus antibodies in pig sera do not necessarily indicate the presence of the tick on the farm where sampling was carried out.