The mechanisms of interactions among carnivore species range from facilitation (mainly through increased availability of prey carcasses) to competition. We assessed the potential for facilitative/competitive interactions between the two most widespread carnivores in the world, the wolf and the red fox, in a prey-rich area recently recolonized by the apex predator. One could expect that the superior competitor would ecologically suppress the inferior one, leading to avoidance of the former by the latter. In a Mediterranean coastal area (2017–2018), we assessed spatiotemporal and dietary interspecific overlap and investigated whether the recovery of wolves affected food habits of foxes. Spatiotemporal overlap was extensive (0.84–0.89). Wild ungulates were the staple of the wolf diet (~88–90%); foxes used mainly invertebrates and fruits (~78%), with ungulates being a substantial food category (13% of diet; 66% of occurrences among vertebrate prey). Interspecific dietary overlap was low (0.23), but extensive (0.89) for vertebrate prey. In comparison to a preceding wolf-free period, the volume and occurrence of large mammals in the diet of foxes showed a 2.8- to 3.5-fold increase. Apparently, foxes did not avoid wolves, which provided additional food to the foxes as prey leftovers. In a rich community, the presence of wolves may increase the food spectrum of foxes. Temporal variation of facilitation vs. competition should be assessed in relationship to spatiotemporal changes of predator–prey numbers.

Simple Summary Wolves and European wildcats are two iconic predator species that can live in overlapping ecological contexts and also share their habitats with their domestic free-ranging relatives, increasing the risk of anthropogenic hybridisation and its possible deleterious consequences. By applying a multidisciplinary approach, we morphologically and molecularly analysed the cat remains found in a canid faecal sample collected in a forested area of central Italy. Individual multilocus genotypes of both predator and prey were identified turning out to be, respectively, a wolf showing traces of dog ancestry at autosomal microsatellite loci and a domestic cat. Abstract Non-invasive genetic sampling is a practical tool to monitor pivotal ecological parameters and population dynamic patterns of endangered species. It can be particularly suitable when applied to elusive carnivores such as the Apennine wolf ( Canis lupus italicus ) and the European wildcat ( Felis silvestris silvestris ), which can live in overlapping ecological contexts and sometimes share their habitats with their domestic free-ranging relatives, increasing the risk of anthropogenic hybridisation. In this case study, we exploited all the ecological and genetic information contained in a single biological canid faecal sample, collected in a forested area of central Italy, to detect any sign of trophic interactions between wolves and European wildcats or their domestic counterparts. Firstly, the faecal finding was morphologically examined, showing the presence of felid hair and claw fragment remains. Subsequently, total genomic DNA contained in the hair and claw samples was extracted and genotyped, through a multiple-tube approach, at canid and felid diagnostic panels of microsatellite loci . Finally, the obtained individual multilocus genotypes were analysed with reference wild and domestic canid and felid populations to assess their correct taxonomic status using Bayesian clustering procedures. Assignment analyses classified the genotype obtained from the endothelial cells present on the hair sample as a wolf with slight signals of dog ancestry, showing a q i = 0.954 (C.I. 0.780–1.000) to the wolf cluster, and the genotype obtained from the claw as a domestic cat, showing a q i = 0.996 (95% C.I. = 0.982–1.000) to the domestic cat cluster. Our results clearly show how a non-invasive multidisciplinary approach allows the cost-effective identification of both prey and predator genetic profiles and their taxonomic status, contributing to the improvement of our knowledge about feeding habits, predatory dynamics, and anthropogenic hybridisation risk in threatened species.

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The gut microbiome can play a fundamental role in several processes associated with an organism’s ecology, and research on the microbiota of wild animals has flourished in the last decades. Microbiome composition can vary across and within species according to taxonomy and environmental variability, including the availability of food resources. Species with a large distribution may exhibit spatial patterns acting at local/regional scales. We considered one of the most widespread and ecologically important predators in the world, i.e., the grey wolf Canis lupus, for which microbiome data is unduly limited. We studied four packs in different ecological conditions in Italy—two packs from a Mediterranean coastal area and two packs from an Alpine range—using an amplicon sequencing barcoding approach. Overall, our results are consistent with food habits entailing a diet largely based on wild prey and agree with findings obtained on other species of canids. If confirmed through a larger sample, they would support the hypothesis of an influence of the shared evolutionary history across canids on the composition of the gut microbiome. Some emerging differences were observed among packs in terms of species composition (Jaccard) and diversity, providing partial support to recent indications on pack identity as a significant determinant of microbiome composition. These results should be considered preliminary results of gut microbiome composition in our study areas.