Abstract

Simple SummaryIt is widely recognized that the assessment of animal welfare should include measures of positive emotional (affective) state. Existing behavioral and physiological indicators of a positive affective state frequently lack sensitivity, objectivity or are unsuitable in a production environment. Therefore, there is a need to develop new approaches to accurately and objectively measure a positive emotional state in animals, including novel molecular markers such a miRNA. These biomarkers must be measurable in the peripheral circulation and provide an accurate account of the physiological and molecular activity in regions of the brain associated with emotional processing. Further, such markers require validation against established behavioral and physiological indices. Here we investigated the efficacy of circulating miRNA as biomarkers of emotional state in the pig.The ability to assess the welfare of animals is dependent on our ability to accurately determine their emotional (affective) state, with particular emphasis being placed on the identification of positive emotions. The challenge remains that current physiological and behavioral indices are either unable to distinguish between positive and negative emotional states, or they are simply not suitable for a production environment. Therefore, the development of novel measures of animal emotion is a necessity. Here we investigated the efficacy of microRNA (miRNA) in the brain and blood as biomarkers of emotional state in the pig. Female Large White × Landrace pigs (n = 24) were selected at weaning and trained to perform a judgment bias test (JBT), before being exposed for 5 weeks to either enriched (n = 12) or barren housing (n = 12) conditions. Pigs were tested on the JBT once prior to treatment, and immediately following treatment. MiRNA and neurotransmitters were analyzed in blood and brain tissue after euthanasia. Treatment had no effect on the outcomes of the JBT. There was also no effect of treatment on miRNA expression in blood or the brain (FDR p > 0.05). However, pigs exposed to enriched housing had elevated dopamine within the striatum compared to pigs in barren housing (p = 0.02). The results imply that either (a) miRNAs are not likely to be valid biomarkers of a positive affective state, at least under the type of conditions employed in this study, or (b) that the study design used to modify affective state was not able to create differential affective states, and therefore establish the validity of miRNA as biomarkers.

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