Muscle tissue from 37 red deer from Norway was examined for sarcocysts. Sarcocysts from 2 reindeer were obtained for comparative studies. Cysts were excised and morphologically classified by light microscopy, scanning electron microscopy, and DNA sequence analysis. Five Sarcocystis species, Sarcocystis hjorti n. sp., Sarcocystis hardangeri, Sarcocystis ovalis, Sarcocystis rangiferi, and Sarcocystis tarandi, were found. All 5 species have previously been identified from either reindeer or moose by their sarcocyst morphology and/or ssu rRNA gene sequence. S. hjorti was the most prevalent species. Multiple variants of the ssu rRNA gene and the first internal transcribed spacer were found in S. rangiferi and S. tarandi from both red deer and reindeer. Phylogenetic analyses indicated that S. tarandi occurs in both red deer and reindeer, but it could not be clearly demonstrated whether the sequence variation within S. rangiferi between hosts was due to different paralogues or/and different species. DNA sequencing was necessary for definitive species identification, since the hair-like protrusions on the cysts of S. hjorti were not always recognizable by light microscopy and since different cervids harbour Sarcocystis species with highly similar cyst morphology of which at least some are not intermediate host specific.
Muscle tissues from 34 moose from Southeastern Norway and two moose from Canada were examined. Sarcocysts were excised and morphologically classified by light microscopy, and some cysts were further examined by scanning electron microscopy or DNA amplification and sequencing at the small subunit (ssu) rRNA gene. In Norwegian moose, three sarcocyst types were recognized, yet five Sarcocystis species were found by sequence analysis. New names were proposed for three species which could be characterised by both morphological and molecular methods, i.e., Sarcocystis alces, Sarcocystis ovalis, and Sarcocystis scandinavica. S. alces was the most prevalent species, whereas S. scandinavica and the two unnamed species were rare and might either use another principal intermediate host or a rare definitive host. The five species in Norwegian moose were different from Sarcocystis alceslatrans isolated from a Canadian moose. Phylogenetic analyses based on complete ssu rRNA gene sequences revealed a close relationship between the six Sarcocystis species from moose and species from reindeer and Sika deer. We conclude that molecular methods are necessary for unequivocal species identification, as different cervid hosts harbour morphologically indistinguishable sarcocysts.
Fresh muscle tissue from six roe deer from Southeastern Norway was examined for sarcocysts. Cysts were excised and morphologically classified by light microscopy, and some cysts were further examined by scanning electron microscopy or DNA amplification and sequencing of the small subunit (ssu) rRNA gene. Two Sarcocystis species, Sarcocystis gracilis and Sarcocystis oviformis n. sp., were found and described by morphological and molecular methods. S. gracilis was found in all animals, whereas S. oviformis was found in only one roe deer. Polymerase chain reaction identification was necessary for definitive species identification, since cysts of S. gracilis varied in surface structure and since cysts of both S. gracilis and S. oviformis were morphologically indistinguishable from sarcocysts in other cervidae. Phylogenetic analyses based on complete ssu rRNA gene sequences revealed a close relationship between S. gracilis and other canine-transmitted Sarcocystis species, whereas S. oviformis formed a well-supported group with Sarcocystis hardangeri of reindeer and Sarcocystis ovalis of moose.
The aim of this study was to determine whether foxes might act as definitive hosts of Sarcocystis alces in moose. In 2 experiments, 6 silver foxes (Vulpes vulpes) and 6 blue foxes (Vulpes lagopus) were fed muscle tissue from moose containing numerous sarcocysts of S. alces, and euthanazed 7-28 days post-infection (p.i.). Intestinal mucosal scrapings and faecal samples were screened microscopically for Sarcocystis oocysts/sporocysts, which were identified to species by means of species-specific primers and sequence analysis targeting the ssu rRNA gene. All foxes in both experiments became infected with Sarcocystis; the oocysts were fully sporulated by 14 days p.i., containing sporocysts measuring 14-15 x 10 microm. Molecular identification revealed that the oocysts/sporocysts belonged to 2 species, S. alces and Sarcocystis hjorti, although sarcocysts of S. hjorti were only identified in moose subsequent to the infection of foxes. In the first experiment, all 8 foxes also became infected with a Hammondia sp. derived from moose, shedding unsporulated, subspherical oocysts, measuring 10-12 microm in diameter, from 6-7 days p.i. onwards. The study proved that canids (the red fox and arctic fox) are definitive hosts for S. alces and S. hjorti, as had been inferred from the phylogenetic position of these species.
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