The severe outbreak of African horse sickness (AHS) in Thailand has forced horses to reside full-time inside barns that are covered by a small mesh net to prevent minuscule AHS insect vectors from gaining access. However, housing in the net-covered barn induces stress in horses, which compromises their welfare. Implementing strategic airflow adjustment while retaining the vector-protection characteristics has been proposed to help alleviate this problem. The present study aimed to investigate the effect of strategic ventilation adjustment on blood cortisol levels, heart rate and behaviour in horses in a vector-protected barn. Nine horses underwent two sequential stabling conditions: vector-protected barn housing and housing in a barn in which the air ventilation was explicitly adjusted. Heart rate was higher in the afternoon in horses housed in the barn without ventilation adjustment, whereas no change was observed in the barn with ventilation adjustment. The vector-protected housing increased the horses’ behavioural scores. Blood cortisol level declined over time, and an earlier decrease was detected at 1400h in the barn with ventilation adjustment. Although airflow adjustment did not appear to statistically alter the stress response in horses during housing in the vector-protected barn, an earlier decline in cortisol level alongside an unchanged heart rate in horses during the day may indicate the positive impact of ventilation adjustment within the vector-protected barn. With limited options to reduce stress or discomfort in horses, this strategic protocol could, at least in part, be applied to managing horses’ welfare during the AHS outbreak.

Culicoides imicola is the main vector for bluetongue (BT) and African horse sickness (AHS) viruses in the Mediterranean basin and in southern Europe. In this study, we analysed partial mitochondrial cytochrome c oxidase subunit I (COI) gene to characterize and confirm population expansion of Culicoides imicola across Spain. The data were analysed at two hierarchical levels to test the relationship between C. imicola haplotypes in Spain (n=215 from 58 different locations) and worldwide (n=277). We found nineteen different haplotypes within the Spanish population, including 11 new haplotypes. No matrilineal subdivision was found within the Spanish population, while western and eastern Mediterranean C. imicola populations were very structured. These findings were further supported by median networks and mismatch haplotype distributions. Median networks demonstrated that the haplotypes we observed in the western Mediterranean region were closely related with one another, creating a clear star-like phylogeny separated only by a single mutation from eastern haplotypes. The two, genetically distinct, sources of C. imicola in the Mediterranean basin, thus, were confirmed. This type of star-like population structure centred around the most frequent haplotype is best explained by rapid expansion. Furthermore, the proposed northern expansion was also supported by the statistically negative Tajima's D and Fu's Fs values, as well as predicted mismatch distributions of sudden and spatially expanding populations. Our results thus indicated that C. imicola population expansion was a rapid and recent phenomenon.