Abstract

BackgroundMonitoring wildlife diseases is needed to identify changes in disease occurrence. Wildlife blood samples are valuable for this purpose but are often gathered haemolysed. To maximise information, sera often go through repeated analysis and freeze-thaw cycles. Herein, we used samples of clean and haemolysed Eurasian wild boar (Sus scrofa) serum stored at -20°C and thawed up to five times to study the effects of both treatments on the outcome of a commercial ELISA test for the detection of antibodies against Suid Herpesvirus 1 (ADV).ResultsThe estimated prevalence of antibodies against ADV was 50-53% for clean and haemolysed sera. Hence, haemolysis did not reduce the mean observed serum antibody prevalence. However, 10 samples changed their classification after repeated freeze-thawing. This included 3 (15%) of the clean sera and 7 (41%) of the haemolysed sera.ConclusionsWe recommend (1) establishing more restrictive cut-off values when testing wildlife sera, (2) recording serum quality prior to sample banking, (3) recording the number of freezing-thawing cycles and (4) store sera in various aliquots to reduce repeated usage. For instance, sera with more than 3 freeze-thaw cycles and a haemolysis of over 3 on a scale of 4 should better be discarded for serum antibody monitoring. Even clean (almost not haemolysed) sera should not go through more than 5 freeze-thaw cycles.

Highlights

  • Monitoring wildlife diseases is needed to identify changes in disease occurrence

  • enzyme-linked immunosorbent assay (ELISA) results coincided in 14 cases (8 positive, 6 negative; 82%)

  • Two negative clean sera tested positive and doubtful, respectively, with haemolysis, and one positive clean sera tested negative with haemolysis

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Summary

Introduction

Monitoring wildlife diseases is needed to identify changes in disease occurrence. Sera often go through repeated analysis and freeze-thaw cycles. We used samples of clean and haemolysed Eurasian wild boar (Sus scrofa) serum stored at -20°C and thawed up to five times to study the effects of both treatments on the outcome of a commercial ELISA test for the detection of antibodies against Suid Herpesvirus 1 (ADV). Monitoring wildlife diseases is needed to identify changes in disease occurrence and to measure the impact of intervention. Wildlife blood samples are often gathered postmortem from shot (hunter-harvested) animals, and centrifuged to obtain serum. Sera are stored frozen and often reused several times in order to maximise the information obtained. Wildlife sera are often haemolysed and/or go through repeated freeze-thaw cycles

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