Sample collection

Sampling of birds was based on purposive sampling technique, i.e., based on availability of birds at the time of capture and purchase. A total of 136 birds, comprising 20 Laughing doves, 22 Speckled pigeons, 25 Cattle egrets, 25 Senegalese parrots, 21 mallards and 23 geese were sampled over a period of 9 months (April-December 2018).
Blood sample was collected from each bird via the wing vein using sterile hypodemic syringes and 23G needles, dispensed into labeled plain sample tubes and allowed to clot at room temperature. Thereafter, the formed serum was transferred into labeled sample bottle and stored at -200C until used for detection of AIV antibodies.
Oropharyngeal and cloacal swabs from each bird were collected using Dacron swab, dispersed in labeled sample bottle containing about 1 ml of virus transport medium (VTM) and pooled together. Pooled swabs of about 4-5 birds of the same species were further pooled together to increase the concentration of antigen for possible detection. The total numbers of pooled oropharyngeal and cloacal swabs for Laughing dove, Speckled pigeon, Senegalese parrot, Cattle egret, mallard and geese were 4, 4, 5, 5, 4 and 4, respectively. The swab and VTM were thoroughly mixed and the mixture of swab content and VTM was stored at -200C until used for viral molecular detection.