Atama Nnomzie C, Chestakova Irina V, de Bruin Erwin, van den Berg Tijs J, Munger Emmanuelle, Reusken Chantal, Oude Munnink Bas B, van der Jeugd Henk, van den Brand Judith M A, Koopmans Marion P G, Sikkema Reina S
Viroscience, Erasmus Medical Center, Rotterdam, the Netherlands.
Vogeltrekstation - Dutch Centre for Avian Migration and Demography, NIOO-KNAW, Wageningen, the Netherlands.
One Health. 2022 Nov 13;15:100456. doi: 10.1016/j.onehlt.2022.100456. eCollection 2022 Dec.
Wild birds are reservoirs of several zoonotic arboviruses including West Nile virus (WNV) and Usutu virus (USUV), and are often monitored as indicators for virus introduction and spread. To optimize the bird surveillance for arboviruses in the Netherlands and to explore the possibilities for citizen science in surveillance, we investigated the suitability of using alternative sample types from live and dead birds. The sensitivity of molecular detection via RT-PCR of viral RNA in feather, heart, lung, throat and cloaca swabs from dead birds, and serum, dried blood spots (DBS) and throat and cloaca swabs from live birds were compared. IgY antibody detection was also assessed from DBS relative to serum on protein-microarray and virus neutralization test. Feathers showed a high detection sensitivity for USUV RNA in both live and dead birds, and no significant decrease was observed in the RNA loads in the feathers after being stored dry at room temperature for 43 days. Additionally, viral RNAs extracted from feathers of day 0 and 43 were successfully sequenced. The results indicated no statistical significant difference in sensitivity and viral loads detection in heart, spleen, and lung relative to corresponding brain samples in dead birds. In live birds, viral RNA loads did not differ between throat and cloaca swabs. This study identified less-invasive sample types that allows involvement of citizens in collecting samples from wild birds for arbovirus surveillance. Sensitivity and specificity of DBS-based antibody detections were significantly lower and therefore need optimization.
野生鸟类是包括西尼罗河病毒（WNV）和乌苏图病毒（USUV）在内的几种人畜共患虫媒病毒的宿主，并且经常作为病毒传入和传播的指标进行监测。为了优化荷兰虫媒病毒的鸟类监测，并探索公民科学在监测中的可能性，我们研究了使用来自活鸟和死鸟的替代样本类型的适用性。比较了通过逆转录聚合酶链反应（RT-PCR）检测死鸟羽毛、心脏、肺、咽喉和泄殖腔拭子以及活鸟血清、干血斑（DBS）、咽喉和泄殖腔拭子中病毒RNA的分子检测灵敏度。还通过蛋白质微阵列和病毒中和试验评估了相对于血清，从DBS中检测IgY抗体的情况。羽毛对活鸟和死鸟中的USUV RNA均显示出高检测灵敏度，并且在室温下干燥储存43天后，羽毛中的RNA含量未观察到显著下降。此外，成功对第0天和第43天羽毛中提取的病毒RNA进行了测序。结果表明，死鸟心脏、脾脏和肺中相对于相应脑样本的检测灵敏度和病毒载量无统计学显著差异。在活鸟中，咽喉和泄殖腔拭子中的病毒RNA载量没有差异。本研究确定了侵入性较小的样本类型，这使得公民能够参与从野生鸟类中采集样本以进行虫媒病毒监测。基于DBS的抗体检测的灵敏度和特异性显著较低，因此需要优化。
