Understanding the impact routine research and laboratory procedures have on animals is crucial to improving their well-being and to the success and reproducibility of the research they are involved in. Cognitive measures of welfare offer insight into animals' internal psychological state, but require validation. Attention bias - the tendency to attend to one type of information over another - is a cognitive phenomenon documented in humans and animals that is known to be modulated by affective state (i.e., emotions). Hence, changes in attention bias may offer researchers a deeper perspective of their animals' psychological well-being. The dot-probe task is an established method for quantifying attention bias in humans (by measuring reaction time to a dot-probe replacing pairs of stimuli), but has yet to be validated in animals. We developed a dot-probe task for long-tailed macaques (Macaca fascicularis) to determine if the task can detect changes in attention bias following anesthesia, a context known to modulate attention and trigger physiological arousal in macaques. Our task included the following features: stimulus pairs of threatening and neutral facial expressions of conspecifics and their scrambled counterparts, two stimuli durations (100 and 1,000 ms), and counterbalancing of the dot-probe's position on the touchscreen (left and right) and location relative to the threatening stimulus. We tested 8 group-housed adult females on different days relative to being anesthetized (baseline and 1-, 3-, 7-, and 14-days after). At baseline, monkeys were vigilant to threatening content when stimulus pairs were presented for 100 ms, but not 1,000 ms. On the day immediately following anesthesia, we found evidence that attention bias changed to an avoidance of threatening content. Attention bias returned to threat vigilance by the third day postanesthesia and remained so up to the last day of testing (14-days after anesthesia). We also found that attention bias was independent of the type of stimuli pair (i.e., whole face vs. scrambled counterparts), suggesting that the scrambled stimuli retained aspects of the original stimuli. Nevertheless, whole faces were more salient to the monkeys as responses to these trials were generally slower than to scrambled stimulus pairs. Overall, our study suggests it is feasible to detect changes in attention bias following anesthesia using the dot-probe task in nonhuman primates. Our results also reveal important aspects of stimulus preparation and experimental design.

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