A non-invasive, improved RIA and overt observation in the study of singleton Apennines’ wolf (Canis lupus) reproductive behavior

The analysis of fecal hormones allows a close but non-invasive monitoring of animals avoidingthe stress of restraint/capture, which in turn can upset animals’ hormonal profile. Steroid hormoneprogesterone was analysed in three singleton, female grey wolves of different age, belonging to theendangered species of the Apennines’ Canis lupus. The analysis was carried out during the breedingseason by using an improved radioimmunoassay on samples collected on the field. To reduce the stressto animals and danger to people, the overt observations were carried out by operators who were alreadyfamiliar with the animals, saving the money of a camera-monitoring-system. Concurrently, a male and afemale gray wolves housed together were monitored as a control. The results indicated the importance ofdehydration of fecal samples before the extraction with petroleum ether, which was shown to be moreefficient than diethyl ether, and that pre-treatment with methanol greatly enhances extraction (p<0.01).Females of Apennines’ grey wolf showed the first sign of oestrus by a vaginal blood loss, that was easilydetected on the ground; the analysis carried out on fecal samples revealed a rapidly declining lutealphase, with P4 metabolites reaching the basal values of a non-cyclic female. In the matter of welfare,behavioural observations on Apennines’ grey wolf showed that unpaired animals, although familiar withthe operators, failed to display a sexual social behavior during the reproductive season, that is thebehavioural signs were hidden in overt observational situation.