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AbstractDomestic turkey production generally utilises only a few genetically improved lines, and local breeds are severely endangered as a result. Furthermore, the genetic resources of domestic turkeys have not been properly investigated, which could, ultimately, lead to the extinction of local breeds and negatively affect their corresponding genetic diversity and environmental adaptation. Although, several microsatellite markers have been designed for mapping and quantitative trait locus analysis, there is no standard panel of markers for genetic characterisation or genetic diversity assessment. Accordingly, the present study aimed to develop a set of polymorphic microsatellite markers that could be used for international turkey population studies. Thirty-nine microsatellites were selected based on polymorphism, DNA sequence and chromosome position, as well as on amplification efficiency, success rate and the absence of nonspecific amplification. The markers were screened using 105 DNA samples from local turkey breeds from Mexico, the United States, Italy, Brazil, Egypt and Spain. A total of 401 alleles were identified, with a mean number of alleles per marker of 10.28 ± 4.25. All microsatellites were polymorphic, with at least four alleles and no more than 19 alleles. Furthermore, allelic richness ranged from 3.810 to 17.985, mean heterozygosity ranged from 0.452 ± 0.229 to 0.667 ± 0.265, polymorphic information content values ranged from 0.213 (MNT264) to 0.850 (RHT0024) and the mean Fis value was 0.322. Overall, the panel was highly polymorphic and exhibited moderate Hardy–Weinberg disequilibrium, thereby indicating its value as a tool for biodiversity and population structure studies that could play an important role in promoting the conservation of local turkey breeds.HighlightsImportant genetic resources reside within indigenous turkey populations. These are linked to historic heritage production values and breeds. It is important to preserve this heritage and genetic diversity, which threatens to be lost as production systems focus on production characteristics.Microsatellite markers, even though, they are now replaced by single nucleotide polymorphism automatic genotyping platforms in many fields of genetics, remain a viable alternative thanks to their cheapness and simplicity of study which makes them particularly useful when the population to be studied lacks information of the prior genetic structure.

With more than 150 recognized breeds, donkeys assume relevant economic importance, especially in developing countries. Even if the estimated number of heads worldwide is 53M, this species received less attention than other livestock species. Italy has traditionally been considered one of the cradles of European donkey breeding, and despite a considerable loss of biodiversity, today still counts nine autochthonous populations. A total of 220 animals belonging to nine different populations were genotyped using the double-digest restriction site associated DNA (ddRAD) sequencing to investigate the pattern of diversity using a multi-technique approach. A total of 418,602,730 reads were generated and successfully demultiplexed to obtain a medium-density SNP genotypes panel with about 27K markers. The diversity indices showed moderate levels of variability. The genetic distances and relationships, largely agree with the breeding history of the donkey populations under investigation. The results highlighted the separation of populations based on their genetic origin or geographical proximity between breeding areas, showed low to moderate levels of admixture, and indicated a clear genetic difference in some cases. For some breeds, the results also validate the success of proper management conservation plans. Identified runs of homozygosity islands, mapped within genomic regions related to immune response and local adaptation, are consistent with the characteristics of the species known for its rusticity and adaptability. This study is the first exhaustive genome-wide analysis of the diversity of Italian donkey populations. The results emphasized the high informativeness of genome-wide markers retrieved through the ddRAD approach. The findings take on great significance in designing and implementing conservation strategies. Standardized genotype arrays for donkey species would make it possible to combine worldwide datasets to provide further insights into the evolution of the genomic structure and origin of this important genetic resource.This is the first study deciphering the genetic structure of Italian donkey populations using SNP data. The results of this study confirmed the history of the investigated populations in terms of both genetic and geographical origins.Donkeys assume relevant economic importance in several countries worldwide. However, the genetic structure of these populations is less investigated compared to other species. The aim of this study was to investigate the genetic background of nine different Italian donkey populations. A total of 220 animals were genotyped with about 27K markers extracted by the double-digest restriction site associated DNA sequencing. The consistency of the results across different approaches agreed with the demographic history, the origin, and previous results on the nine donkey populations, suggesting that our conclusions are robust. Moreover, the results of the present study highlighted low to moderate levels of admixture and, for some breeds, confirmed the success of proper management conservation plans.