Changes in social and feeding behaviors, activity, and salivary serum amyloid A in cows with subclinical mastitis

The aim of this study was to identify detailed changes in behavior, and in salivary serum amyloid A (SAA), associated with subclinical mastitis. This included standard sickness behaviors, such as decreased activity, feeding and drinking (here labeled “core maintenance” behaviors), and less well-studied social, grooming, and exploratory behaviors (here labeled “luxury” behaviors). Luxury behaviors are biologically predicted to change at lower levels of mastitis infection and are, therefore, particularly relevant to detecting subclinical mastitis. Salivary serum amyloid A is a physiological marker of systemic inflammation, with levels in milk and serum already known to increase during subclinical mastitis. We investigated whether the same was true for SAA in cow saliva. Data were collected for 17 matched pairs of commercial barn-housed Holstein-Friesian cows. Each pair comprised a cow with subclinical mastitis (SCM) and a healthy control (CTRL), identified using somatic cell count (SCC; SCM: SCC >200 × 1,000 cells/mL; CTRL: SCC <100 × 1,000 cells/mL). SCM cows were selected for study ad hoc, at which point they were paired with a CTRL cow, based upon parity and calving date; consequently, the full data set was accrued over several months. Data were collected for each pair over 3 d: SCC (d 1), behavior (d 2), salivary SAA (d 3). All behaviors performed by the focal cows over a single 24-h period were coded retrospectively from video footage, and differences between the SCM and CTRL groups were investigated using the main data set and a subset of data corresponding to the hour immediately following morning food delivery. Saliva was collected using cotton swabs and analyzed for SAA using commercially available ELISA kits. We report, for the first time, that an increase in salivary SAA occurs during subclinical mastitis; SAA was higher in SCM cows and demonstrated a positive (weak) correlation with SCC. The behavioral comparisons revealed that SCM cows displayed reductions in activity (behavioral transitions and distance moved), social exploration, social reactivity (here: likelihood to be displaced following receipt of agonism), performance of social grooming and head butts, and the receipt of agonistic noncontact challenges. In addition, SCM cows received more head swipes, and spent a greater proportion of time lying with their head on their flank than CTRL cows. The SCM cows also displayed an altered feeding pattern; they spent a greater proportion of feeding time in direct contact with 2 conspecifics, and a lower proportion of feeding time at self-locking feed barriers, than CTRL cows. Behavioral measures were found to correlate, albeit loosely, with serum SAA in a direction consistent with predictions for sickness behavior. These included positive correlations with lying duration and the receipt of all agonistic behavior, and negative correlations with feeding, drinking, the performance of all social and all agonistic behavior, and social reactivity. We conclude that changes in salivary SAA, social behavior, and activity offer potential in the detection of subclinical mastitis and recommend further investigation to substantiate and refine our findings.