Typically, carnivores serve as definitive hosts for Sarcocystis spp. parasites; currently, their role as intermediate hosts is being elucidated. The present study aimed to identify and molecularly characterize Sarcocystis cysts detected in striated muscle of red foxes from different populations in Latvia, Lithuania and Spain. Muscle samples from 411 red foxes (Vulpes vulpes) and 269 racoon dogs (Nyctereutes procyonoides) from Latvia, 41 red foxes from Lithuania and 22 red foxes from Spain were examined for the presence of Sarcocystis sarcocysts by light microscopy (LM). Sarcocystis spp. were identified by transmission electron microscopy (TEM) and molecular biology techniques. Sarcocystis cysts were detected in 11/411 (2.7%) Latvian, 3/41 (7.3%) Lithuanian, and 6/22 (27.3%) Spanish red foxes, however, cysts were not observed in the muscles of racoon dogs. Based on LM, TEM, 18S rDNA, 28S rDNA, ITS1, cox1 and rpoB sequences, Sarcocystis arctica and Sarcocystis lutrae cysts were identified in red fox muscles from Latvia and Lithuania, whereas only S. arctica was detected in Spain. The 18S rDNA, 28S rDNA and ITS1 sequences from the 21 isolates of S. arctica from Latvia, Lithuania and Spain were identical. By contrast, two and four haplotypes were determined based on mtDNA cox1 and apicoplast rpoB sequences, respectively. Polymorphisms were not detected between the two isolates of S. lutrae from Latvia and Lithuania. Based on phylogenetic results, S. arctica and S. lutrae were most closely related to Sarcocystis spp. using predatory mammals as intermediate hosts and to Sarcocystis species with a bird-bird life-cycle. Based on current knowledge, the red fox and Arctic fox (Vulpes lagopus) could act as intermediate host for the same two Sarcocystis species. Molecular results suggest the existence of two genetic lineages of S. arctica, and such divergence relies on its geographical distribution but not on their intermediate host species.

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The Persian fallow deer (Dama dama mesopotamica) formerly widespread in the Middle East was described scientifically at the end of the 19th century and considered extinct ever since. In 1956 it was rediscovered in south-western Iran. As a result, several countries have undertaken actions to reintroduce this subspecies in its native territory. In 2007 the Christian Oswald Foundation, in close cooperation with Iranian institutions, launched plans of in situ and ex situ breeding actions, with its centre in the German Von Opel Zoo and with cooperative Mediterranean partner countries as Israel, to support conservation efforts under scientific control. We performed genetic studies to study the suspected hybridization with European fallow deer (Dama dama dama) and a commitment to preserve pureblood populations. We used a set of microsatellite loci to examine genetic variation and recent hybridization with the European fallow deer. All microsatellite loci used were polymorphic, but some were monomorphic within subspecies. The allelic richness was similar in both subspecies but the ‘private allelic richness’ was reduced to a half in the Persian fallow deer, signalling allelic loss due to genetic drift and inbreeding. Moreover, we showed the presence of two discrete groups representing the two subspecies, with no signs of admixture or hybridization. Furthermore, Persian fallow deer studied here belong to two pre-defined genetics groups: the wild and the (more genetically impoverished) captive populations of Persian fallow deer. Finally, the Persian fallow deer deserves a high conservation priority, both in the Iranian stock and in the captive populations, so as to avoid hybridization.