2.1 Samples
The sampling was carried out by Local Official Veterinary Service during Official Eradication Control Plane as specified in the Regulation (Eu) 2016/429 of the European Parliament and of the Council of 9 March 2016 on transmissible animal diseases and amending and repealing certain acts in the area of animal health (‘Animal Health Law’). The permission was obtained from the owners of farm animals for collection of their nasal and rectal swab specimens. All methods were performed in accordance with the relevant guidelines and regulations.
The farm included 150 animals, of which 24 lactating cows were sampled (Table 1). The geographical location of the herd was mapped using a geographic information system (GIS) (Figure 1). The herd had never been vaccinated against BCoV.
All samples were collected by local veterinary authorities. Nasal and rectal swab specimens were collected, frozen, and stored at -80°C until processing. Blood was drawn from the jugular vein using sterile evacuated tubes without EDTA anticoagulant. Upon arrival at the laboratory, the samples were centrifuged, and aliquots of the serum and milk were frozen at −80°C until testing.