Members of the family Corvidae are ecologically flexible omnivorous birds, particularly adaptive to urban habitats, and living in proximity to humans; these birds may serve as definitive hosts (DH) for Sarcocystis spp., but research about this is lacking. In the present study, intestinal samples from 91 corvids collected in Lithuania were molecularly tested by species-specific PCR targeting the ITS1 and cox1 genes and subsequently sequenced for the presence of Sarcocystis spp. Under a light microscope, oocysts of Sarcocystis spp. were observed in 43 samples (47.3%), while molecular methods, detected Sarcocystis spp. in 77 birds (84.6%). Eleven Sarcocystis spp. (S. columbae, S. cornixi, potentially pathogenic S. halieti, S. kutkienae, S. lari, S. turdusi, S. wobeseri, S. arctica, S. lutrae, S. ovalis, and S. oviformis) were identified in the intestinal samples from six corvid species from Lithuania. Infections with multiple Sarcocystis spp. were detected in 79.2% of the infected corvid birds. Three of the identified Sarcocystis spp. use corvids as intermediate hosts (IH); therefore, corvids may serve as IH and DH of the same Sarcocystis species. Based on molecular results and on corvid diet, omnivorous corvids may play an important role in transmitting Sarcocystis spp.

Background: Birds of the family Laridae have not been intensively examined for infections with Sarcocystis spp. To date, sarcocysts of two species, S. lari and S. wobeseri, have been identified in the muscles of gulls. The aim of the present study was to evaluate the species richness of Sarcocystis in the herring gull, Larus argentatus, from Lithuania. Methods: In the period between 2013 and 2019, leg muscles of 35 herring gulls were examined for sarcocysts of Sarcocystis spp. Sarcocystis spp. were characterised morphologically based on a light microscopy study. Four sarcocysts isolated from the muscles of each infected bird were subjected to further molecular examination. Sarcocystis species were identified by means of ITS1 sequence analysis. Results: Sarcocysts were detected in 9/35 herring gulls (25.7%). Using light microscopy, one morphological type of sarcocysts was observed. Sarcocysts were microscopic, thread-like, had a smooth and thin (about 1 µm) cyst wall and were filled with banana-shaped bradyzoites. On the basis of ITS1 sequences, four Sarcocystis species, S. columbae, S. halieti, S. lari and S. wobeseri, were identified. Furthermore, it was demonstrated that a single infected herring gull could host two Sarcocystis species indistinguishable under light microscopy. Conclusions: Larus argentatus is the first bird species found to act as intermediate host of four Sarcocystis spp. According to current knowledge, five species, S. falcatula, S. calchasi, S. wobeseri, S. columbae and S. halieti can use birds belonging to different orders as intermediate hosts.

One morphologic type of sarcocyst was found in 26% (7/27) of Eurasian Coots (Fulica atra) and was described as Sarcocystis fulicae n. sp. using morphologic, 18S ribosomal (r) DNA, 28S rDNA, and internal transcribed spacer 1 (ITS1) analysis. By light microscope, cysts were ribbon-shaped and measured 7.3 mm in length by 116 mu m wide. Histologically, the cyst wall reached up to 1.2 lm in thickness and seemed smooth. The detected sarcocysts were packed with relatively small banana-shaped bradyzoites that were 6.7x2.0 lm in size. Ultrastructurally, the cyst wall amounted to 1 lm and had many conical protrusions but seemed almost smooth in some places. The parasitophorous vacuolar membrane appeared undulating, with knob-like blebs and invaginations. The cyst wall belonged to type 1d. Morphologically, cysts of S. fulicae differed considerably from cysts of Sarcocystis atraii previously described in the same host but were indistinguishable from those of Sarcocystis corvusi, Sarcocystis lari, and Sarcocystis wobeseri, which use birds as intermediate hosts. According to the phylogenetic and ecologic data, birds of prey, mostly the Western Marsh Harrier (Circus aeruginosus) and the White-tailed Eagle (Haliaeetus albicilla), are presumed to be definitive hosts of S. fulicae.

Data on the distribution of different Sarcocystis species in various muscles of sheep are scarce. In the present study, 190 diaphragm, oesophagus, and heart muscle samples of 69 sheep raised in Lithuania were examined for the presence of Sarcocystis spp. Under a light microscope, two morphological types of microcysts corresponding to S. arieticanis and S. tenella were detected. Eight and 12 sarcocysts of S. arieticanis and S. tenella, respectively, were isolated and characterised by the sequencing of a portion of cox1. The sequence comparisons revealed the highest similarity between European and Asian isolates of S. arieticanis and S. tenella obtained from domestic sheep and other wild Caprinae hosts. Based on peptic digestion, nested PCR targeting cox1, and sequencing, a 100% infection prevalence of S. arieticanis and S. tenella was observed in the 69 studied animals. The occurrence of S. tenella was significantly higher in the diaphragm than in the oesophagus (χ2 = 13.14, p < 0.001), whereas differences in the prevalence of S. arieticanis in the studied muscle types were insignificant (χ2 = 1.28, p > 0.05). Further molecularly based epidemiological studies are needed to compare the prevalence of Sarcocystis species in various muscles of sheep raised in different geographic regions.

Background: Members of the genus Sarcocystis are protozoan parasites characterized by a prey–predator two-host life-cycle. Sarcocysts are formed in the muscles or central nervous system of the intermediate host (IH), while sporocysts develop in the small intestine of the definitive host (DH). Various birds of prey have been confirmed to be DH for Sarcocystis spp. Three Sarcocystis species, S. wobeseri, S. halieti and S. falcatula, have been identified in the muscles of birds of prey, of which the latter are known to be pathogenic and can cause encephalitis in various birds. The aim of this study was to identify Sarcocystis spp. in the muscles of birds of prey from Spain. Methods: Between 2019 and 2020, muscle tissue samples taken from 59 birds of prey admitted to the Wildlife Recovery Centre in Ilundain (Navarra, Spain) were examined for the presence of Sarcocystis spp. Sarcocysts in fresh squashed samples were morphologically characterized under the light microscope (LM). Sarcocystis spp. were identified by means of 28S ribosomal RNA and internal transcribed spacer 1 sequence analysis. Results: Microscopic examination of squashed tissue samples stained with methylene blue revealed the presence of sarcocysts in three of the 59 (5.1%) birds examined. Only one sarcocyst type was observed under the LM. Sarcocysts were thread-like (1050–2160 × 130–158 μm) and had a thin (0.7–1.4 μm) and smooth cyst wall. Septa divided the cysts into compartments filled with banana-shaped (5.9 × 1.7 μm) bradyzoites. On the basis of DNA sequence results, S. halieti was identified in the western marsh harrier (Circus aeruginosus) and the black kite (Milvus migrans) for the first time. Sarcocysts of S. halieti were shorter and wider compared to those observed in the great cormorant (Phalacrocorax carbo) and the herring gull (Larus argentatus). According to current knowledge, S. halieti may infect birds belonging to four different orders: Suliformes, Charadriiformes, Strigiformes and Accipitriformes. Conclusions: This is the first report of S. halieti in the western marsh harrier and the black kite as IH. So far, little research has been conducted on birds of prey as IH for Sarcocystis spp. These results indicate that further studies combining morphological, histopathological, and molecular methods are required. Graphical

There is a lack of research on the role of mustelids in the transmission of various Sarcocystis spp. In the present study we tested the hypothesis that widespread mustelids in Lithuania could be involved in the transmission of Sarcocystis spp. using cattle as intermediate hosts. In 2016–2020, intestinal samples of 84 mustelids were examined. Sarcocystis spp. were identified by species-specific PCR targeting the cox1 gene and subsequent sequencing. Under a light microscope, oocysts/sporocysts of Sarcocystis spp. were observed in 40 samples (47.6%), while using molecular methods, they were detected in 75 animals (89.3%). Four Sarcocystis spp. were identified in the intestinal samples of American mink (Neovisonvison), Beech marten (Martes foina), European pine marten (Martes martes), European badger (Meles meles) and European polecat (Mustela putorius). The prevalence of predominant Sarcocystis spp., S. bovifelis (89.3%) and S. cruzi (73.8%) was significantly higher than that of S. hirsuta (3.6%) and S. hominis (1.2%). In an individual sample, most frequently two Sarcocystis spp. were identified (69.0%), then a single species (15.5%) and three species (4.8%). The present study provides strong evidence that mustelids serve as definitive hosts for Sarcocystis spp. using cattle as intermediate hosts.