# Genetic Monitoring of a Newly Established Grey Wolf Population in a Peri-Urban Protected Area with First Insights into Wolf–Dog Hybridization in Greece

## TL;DR

This study genetically monitors a newly established grey wolf population in Greece and finds no evidence of wolf-dog hybridization in the area.

## Contribution

The first systematic genetic monitoring of grey wolves in Greece and insights into potential hybridization in northern regions.

## Key findings

- A minimum of 31 unique wolves were identified, structured into at least three packs.
- No wolf–dog hybrids were detected in the Parnitha Protected Area using microsatellite and SNP analyses.
- A possible hybrid was found among reference samples from Northern Greece with atypical traits.

## Abstract

Background/Objectives: Following centuries of systematic eradication, grey wolf (Canis lupus) populations across Europe have experienced a significant recovery over recent decades, which leads to concerns regarding, among others, anthropogenic hybridization. In Greece, the genetic status of the wolf population is largely unknown to date. Here, we genetically monitor and test for wolf–dog hybridization events in a recently established wolf population in the Parnitha Protected Area, in close vicinity to the capital city of Greece. Methods: One hundred and twenty-four wolf scat samples were genotyped at 20 canine-specific autosomal microsatellite loci and compared to available reference tissue samples from wolves and free-ranging dogs. Results: A minimum of 31 unique wolf individuals were identified, structured into at least three packs. No wolf–dog hybrids were detected in the study area. To validate the accuracy of the microsatellite analysis, an ancestry informative 93-SNP panel was applied to non-invasive wolf DNA samples from the study area, confirming the absence of hybrids among them. However, a possible wolf–dog hybrid was detected among reference wolf samples collected in Northern Greece, where individuals with atypical morphological traits are observed. The estimated census population size was in accordance with concurrently obtained camera trapping data, while heterozygosity values were low. Conclusions: This research represents the first systematic effort in Greece to genetically monitor wolves recently established in a protected area. It highlights the need for targeted management strategies based on genetic data to ensure balanced long-term conservation of wolves in peri-urban areas.