Genetic Diversity Analysis of Mitochondrial Cytb Gene, Phylogeny and Phylogeography of Protected Griffon Vulture (Gyps fulvus) from Serbia

Abstract

Once a widespread and common species across the region of southeast Europe, the Griffon vulture is now confined to small and isolated populations across the Balkan Peninsula. The population from Serbia with 290 couples represents its biggest and most viable population that can serve as an important reservoir of genetic diversity from which the birds can be used for the region’s reintroduction or recolonization programs. To estimate the level of genetic diversity, the mitochondrial Cytb gene from 58 unrelated birds sampled during the marking in the nests was sequenced and compared to the homologous Griffon vulture sequences available in publicly accessible online databases. Phylogeographic analysis based on Cytb sequences showed that the most frequent haplotype is found in all Griffon vulture populations and that each population possesses private haplotypes. Our data suggest that the Griffon vulture population from Serbia should be used as a source population for restocking and reintroduction programs in the region. The observed genetic differentiation between the populations from the Iberian and Balkan Peninsulas suggest that the introduction of foreign birds from remote populations should be avoided and that birds from indigenous or neighboring populations, if available, should be used instead.

Sex ratio and relatedness in the Griffon vulture (Gyps fulvus) population of Serbia

“…The Serbian Griffon vulture population experienced a similar fate as other populations in Europe in the form of serious population decline. Although it made it prone to drift and bottleneck effects this population retained significant genetic variability similar to those detected in stable and large populations ( Davidovic et al, 2020 ; Davidovic et al, 2022 ). Nevertheless, there is always a concern about the relatedness and kinship of remaining birds due to their natal philopatric behaviour and inbreeding imprint on the population’s overall fitness.…”

“…No. K0721; Thermo Fisher Scientific, Waltham, MA, USA) and a modified salting-out protocol as described in Davidovic et al (2022) . The quality of the DNA extracts was checked both by the spectrophotometer (NanoPhotometer, IMPLEN, Westlake Village, CA, USA) and agarose gel electrophoresis.…”

“…Mitochondrial haplotypes were determined by comparison with the mtDNA reference sequence for Griffon vulture, ( Mereu et al, 2017 ), following the nomenclature that includes the nucleotide position and adequate substitution different to the reference mitogenome. Cytb sequences used in this analysis were published in Davidovic et al (2022) and can be accessed by the following accession numbers: , , , , , , , , and . Newly sequenced D-loop I sequences were deposited in GenBank under the following accession numbers: – .…”

“…A study by Davidovic et al (2020) showed that this population’s genetic variability (based on the variability of 10 microsatellites loci) was similar to numerous, unscathed populations such as the Iberian Griffon vulture population and therefore is proposed to serve as a stock population for further reintroduction efforts in the Balkan Peninsula. In addition, analysis of the mitochondrial DNA (mtDNA) gene, Cytb , further demonstrated that the Griffon vulture population of Serbia exhibits substantial levels of genetic diversity as well as the presence of private haplotypes which stresses the need of using the local birds in the future reintroduction programmes planned for the region ( Davidovic et al, 2022 ). However, determining the status of the endangered population under conservation measures should be based on more than just 1 or 2 parameters of genetic variability.…”

“…Breeding parameters in the Serbian Griffon vulture population are relatively high despite the substantial increase in population size, implying that the population is far from its saturation level ( Marinkovic et al, 2021 ) further strengthening this population prospect as a reintroduction resource in the Balkan Peninsula ( Davidovic et al, 2020 , 2022 ). The latest research on the spatial ecology of Griffon vultures in the Balkans identified seven key vulture zones (one in Serbia, one shared between North Macedonia and Bulgaria, one shared between Bulgaria and Greece, two in Bulgaria, one in western Greece and one shared between Kvarner Archipelago islands in Croatia and the Julian Alps—Italy, Austria and Slovenia) ( Peshev et al, 2021 ).…”