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The Middle Ages represented a crucial period for the evolutionary history of wolves (Canis lupus), marked by both significant ecosystem changes, especially through the degradation of wooded landscapes and heavy persecution, that drove this species to a dramatic demographic decline. In Europe, informative and well-documented wolf remains from the Medieval Ages are exceptionally rare and are mostly represented by teeth and postcranial elements. In this study, we describe a well-preserved wolf cranium dated to ca. 967–1157 AD from the Po Valley (northern Italy). The specimen was analysed through a multidisciplinary approach including CT-based, ancient DNA, and palaeopathological analyses. Morphological and genetic data supported the assignment of this sample to Canis lupus species. CT-based analyses indicated a typical wolf-like morphology falling into the extant variability of the medium-sized subspecies C. lupus italicus, whereas palaeopathological analyses indicated a severe periodontitis. Phylogenetic analyses showed that the Po valley wolf had a unique and never described mtDNA control region haplotype, testifying variability in the ancient Italian wolf, which has now been lost. This study provides the first comprehensive description of a wolf from the Middle Ages, adding useful information for a deeper knowledge about population dynamics, variability, and diseases of this species.

Hybridization is a natural or anthropogenic process that can deeply affect the genetic make-up of populations, possibly decreasing individual fitness but sometimes favoring local adaptations. The population of Italian wolves (Canis lupus), after protracted demographic declines and isolation, is currently expanding in anthropic areas, with documented cases of hybridization with stray domestic dogs. However, identifying admixture patterns in deeply introgressed populations is far from trivial. In this study, we used a panel of 170,000 SNPs analyzed with multivariate, Bayesian and local ancestry reconstruction methods to identify hybrids, estimate their ancestry proportions and timing since admixture. Moreover, we carried out preliminary genotype–phenotype association analyses to identify the genetic bases of three phenotypic traits (black coat, white claws, and spur on the hind legs) putative indicators of hybridization. Results showed no sharp subdivisions between nonadmixed wolves and hybrids, indicating that recurrent hybridization and deep introgression might have started mostly at the beginning of the population reexpansion. In hybrids, we identified a number of genomic regions with excess of ancestry in one of the parental populations, and regions with excess or resistance to introgression compared with neutral expectations. The three morphological traits showed significant genotype–phenotype associations, with a single genomic region for black coats and white claws, and with multiple genomic regions for the spur. In all cases the associated haplotypes were likely derived from dogs. In conclusion, we show that the use of multiple genome-wide ancestry reconstructions allows clarifying the admixture dynamics even in highly introgressed populations, and supports their conservation management.

During historical times many local grey wolf (Canis lupus) populations underwent a substantial reduction of their sizes or became extinct. Among these, the wolf population once living in Sicily, the biggest island of the Mediterranean Sea, was completely eradicated by human persecution in the early decades of the XX century.In order to understand the genetic identity of the Sicilian wolf, we applied ancient DNA techniques to analyse the mitochondrial DNA of six specimens actually stored in Italian museums.We successfully amplified a diagnostic mtDNA fragment of the control region (CR) in four of the samples. Results showed that two samples shared the same haplotype, that differed by two substitutions from the currently most diffused Italian wolf haplotype (W14) and one substitution from the only other Italian haplotype (W16). The third sample showed a wolf-like haplotype never described before and the fourth a haplotype commonly found in dogs.Furthermore, all the wolf haplotypes detected in this study belonged to the mitochondrial haplogroup that includes haplotypes detected in all the known European Pleistocene wolves and in several modern southern European populations.Unfortunately, this endemic island population, bearing unique mtDNA variability, was definitively lost before it was possible to understand its taxonomic uniqueness and conservational value.

Mortality due to illegal killing is still today one of the main threats to wolf conservation, and an effective management of the conflict between wolf presence and husbandry is a key element for species conservation. The research was aimed at identifying the farm characteristics and the environmental factors that influence predation, and at formulating predictive models of predation risk. We collected and analysed the data on official predation events that occurred during the period 2005-2012 in an area of the northern Apennines, and on the characteristics of livestock farms recorded at the veterinary services. Furthermore, we mapped the grazing areas used by livestock farms and measured 23 variables of the pastures. Our results showed that the majority of predation events were upon cattle, and that grazing management significantly influences the number of predation events. In particular, the pastures that suffered predation were those in which births occur directly on the pasture, those that had at least one period of free grazing during the year, and cattle farms that were lacking in any preventive methods. The number of killed animals per event was higher for sheep than for cattle and goats and increased progressively during the study period. Predation risk increases if the farms practice free grazing at least for a period during the year, if they are not protected by any preventive method and if the degree of surveillance is regular or constant. The risk of predation also increases with the increasing complexity of pasture shape, the decrease in the percentage of coniferous forest and if the pasture is exposed to the north. The model of predation risk showed that 56.6% of the pastures in the study area are potentially exposed to wolf predation, and it allowed us to identify the grazing areas where it is worthwhile intervening with preventive methods.

One hundred fifteen samples of wolf (Canis lupus) feces were collected during 1994 to 1995 from four free-living populations of the north central Apennines Mountains, Italy. The samples were tested for canine parvovirus by antigen-capture enzyme-linked immunosorbent assay (ELISA), hemagglutination, and virus isolation. Four of these samples were positive by virus isolation as confirmed by electron microscopy. All positive samples were from Casentino Park in Tuscany. This is the first definitive observation of canine parvovirus in wolves from Europe.