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Little is known about the relatedness structure of carnivores living in urban areas, where green spaces may vary in size and resource availability. We examined the minimum population size, relatedness structure, and genetic diversity of a recently established population of eastern coyotes (Canis latrans) inhabiting New York City (NYC). The population has been established for approximately 25 years, and sample collection for genetic analysis has been ongoing since 2010. We genotyped 234 scat, eight tissue, and three blood samples at nine microsatellite loci. We identified 45 individual coyotes with a male-biased sex ratio of 2.2:1. We also found moderate to high levels of genetic diversity, with average observed heterozygosity of 0.779 and mean number of alleles per locus of 7.8. Most of the green spaces surveyed supported a single group of closely related coyotes in each. Relatedness comparisons between parks also indicated that coyotes compared across different parks were also closely related. We identified two unrelated mated pairs and found no support for polygamy. The high incidence of relatedness suggests that the coyote population is descended from a small number of founding individuals. Additionally, we genetically recaptured several coyotes, including one individual sampled in the Bronx and in Queens, with a median of 103 days between resampling. This result indicates that the coyotes are persisting in some of the isolated greenspaces of New York City and able to move successfully between them.

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