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In 2016, a study of mDNA sequences of both modern and ancient wolves indicated that in Europe, the two most genetically distinct haplotypes form the Iberian wolf and separately the Italian wolf. The phylogenetic tree generated from the sequences showed the Italian wolf positioned close to the ancient wolves of the Late Pleistocene. In 2017, a study found a second mDNA haplotype that belonged to the Italian wolf, and called for the morphologically and genetically distinct Italian wolf to be considered as a subspecies.

In 2019, an mDNA study of 19 Late Pleistocene-Holocene wolf samples from northern Italy found that these fell within mitochondrial haplogroup 2 except for one sample. Four out of the six detected haplotypes matched ancient Beringian wolves, ancient wolves from northern Europe, some modern European and Chinese wolf populations, and are closely related to the two haplotypes currently found in the Italian wolves. The Italian wolf haplotypes were only one or two mutations away from those of the Pleistocene wolves, indicating mutation in their Italian glacial refuge. The Italian wolf population represents genetic uniqueness highlighted in several mitochondrial and nuclear DNA studies. It is the only remaining wolf population in Europe which belongs exclusively to an mDNA haplogroup that was once widespread in central and western Europe for over 40,000 years, and in North America until the Last Glacial Maximum. Additionally, one canid specimen from the Cava Filo archaeological site of San Lazzaro di Savena, Bologna fell within the domestic dog clade A haplotype — it was radio-carbon dated to be 24,700 years old.
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Evolution of the wolf
A description of the superficial brain morphology of jackals (C. mesomelas, C. aureus), coyotes (C. latrans), wolves (C. lupus, C. rufus), and dogs indicated that the cerebellum of the dog closely approximates that of the coyote, which is closely aligned with the jackals, and that the wolves show numerous brain traits distinct from the other species (Atkins and Dillon 1971). Wolves also have serological and biochemical traits distinct from dogs (Leone and Wiens 1956; Lauer, Kuyt & Baker 1969).

During the Last Glacial Maximum, there was greater wolf genetic diversity than there is today, and within the Pleistocene gray wolf population the variations between local environments would have encouraged a range of wolf ecotypes that were genetically, morphologically and ecologically distinct from one another. One author has proposed that the most likely explanation for the different morphological characteristics of the dog compared to the wolf is that the dog’s ancestor was adapted to a different niche than the wolf.

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