Designing an assay to evaluate behavioral responses to opposite‐sex conspecifics in the endangered black‐footed ferret (Mustela nigripes)

The endangered black‐footed ferret (ferret; Mustela nigripes) is a North American carnivore that is actively managed to reestablish self‐sustaining wild populations. Behavioral abnormalities have been reported in the breeding program and may be a limiting factor for the species' success. Our goal was to design and test an assay that examines the ferret's exploratory response to odor cues in the form of soiled bedding from opposite‐sex conspecifics. Across two breeding seasons, males and females were tested using a T‐maze that connected their home nest box to two novel nest boxes containing two different conspecific's soiled bedding. For a control, we provided two clean bedding samples. We ran linear mixed models to determine the effect of sex, type of odor cue (soiled, clean), and order of trial (first, second) on time exploring and proportion of that time spent in each behavior. Ferrets spent the majority of time in the novel nest boxes sniffing (44%), standing alert (27%) and scratching (14%). Males explored for longer than females; however, both displayed similar behaviors. Type of cue influenced behavior, with ferrets sniffing more among soiled cues than clean cues. Habituation to the assay was also observed, with less exploration and more standing alert during the second trial of the day. This study is the first step in characterizing the ferret's exploratory response and provides information regarding vital investigatory and vigilance behaviors. The continual development of this assay to further evaluate reproductive and mate choice behaviors will facilitate more successful breeding of the species. A modified T‐maze was used to observe olfactory exploration and communication among fossorial black‐footed ferrets. Male and female ferrets explored the two novel nest boxes and sniffed soiled odor cues from different opposite sex conspecifics more than clean control cues. Research Highlights T‐maze allows ferret‐specific behaviors to be observed. The first trial and soiled bedding cue influenced exploratory behavior and activity budget. Males explored for longer, but both sexes displayed similar behaviors.