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Sarcocystis spp. are cysts-forming coccidian parasites which infect several animals including camels as intermediate hosts. The present study was designed to study the Sarcocystis infection in camels (Camelus dromedaries) at morphological and molecular levels. Sample of the esophageal, heart and ocular muscles tissue were collected from infected camels from El-kharga (New valley Governorate) and examined for Sarcocystis spp. using macroscopic evaluation, light microscopy (LM), transmission electron microscopy (TEM) and molecular analysis. Two species Sarcocystis cameli and Sarcocystis ippeni were recognized. Sarcocysts were thin walled with barely visible projections. Using the TEM, the two structurally distinct sarcocysts were recognized by unique villar protrusions (Vp). Sarcocysts of S. cameli had Vp of type 9j. The sarcocyst wall had upright slender Vp, up to 0.1-0.4 μm long and 0•05-0.1 μm wide. On each Vp, Rows of knob-like protrusions are present appeared to be interconnected. Sarcocystis ippeni had ‘‘type 32’’ sarcocyst wall characterized by conical Vp with an electron dense knob. The Vp were approximately 175.27-266.76 nm long, 100.45-175.98 nm wide. In each Sarcocystis, the Vp had microtubules (Mt) that originated at midpoint of the ground substance (Gs) and continued up to the tip. Molecular data revealed the amplification of partial fragments of the 18S rRNA gene (~600 bp). The digestion analysis of obtained PCR products using RFLP method utilizing (Mbo1) appeared 2 bands approximately 250, 350 bp for each Sarcocystis. Molecular analysis demonstared the ability of 18S rRNA gene for distinguishing Sarcocystis species in studied animals and to be used as molecular marker.

Previous morphological studies suggested that mouflon may have sarcocysts similar to those of sheep. However, to date, no molecular-based studies of the species of Sarcocystis infecting mouflon have been done. The present study identified Sarcocystis species in diaphragm muscle samples from 20 European mouflon (Ovis gmelini musimon). Molecular identification using the cox1 sequence analysis was performed on sarcocysts excised from muscle tissue and on DNA from digested muscle samples. Both frequency and intensity of infection in mouflon were high with 19 of 20 animals testing Sarcocystis positive and > 50 cysts per gram of tissue recovered from 10 of the 19 Sarcocystis positive animals. Molecular analysis revealed dominant Sarcocystis tenella (18/19 animals) and Sarcocystis arieticanis (1/19 animals), whose known intermediate hosts are sheep. In addition, Sarcocystis capracanis, which is known to form sarcocysts in goats, was detected in two animals. The results of this study demonstrated the digestion method to be superior over the direct isolation of sarcocysts for the molecular identification of Sarcocystis species in a certain host. Future research of Sarcocystis diversity in wild ovine and caprine species is needed.

Sarcocystis is an intracellular protozoan parasite in the phylum Apicomplexa. It is widely distributed all over the world. There are scarce reports about chicken Sarcocystis. From February 2016 to January 2018, a total number of 630 chicken carcasses, intestines and viscera were collected from different chicken markets in Menoufia and Gharbia Governorates, Middle region of the Nile Delta, Egypt and carefully inspected. Macroscopic and microscopic cysts of Sarcocystis spp. were found in the intestinal wall and mesentery of 5 birds. Histopathological sections revealed the presence of two shapes of the macroscopic cysts (oval and kidney shape). Their wall was striated and characterised by the presence of radial septa. It had compartments mostly of hexagonal shape, containing both bradyzoites and metrocytes in the periphery. The bradyzoites were banana-shaped and measured 20–30 × 8–10 μm with centrally or posteriorly located nuclei. Microscopic cysts of Sarcocystis spp. were detected in-between muscle bundles, with variable shapes (spindle and oval).

This study aimed to identify Sarcocystis species isolated from macroscopic sarcocysts from naturally infected domestic sheep and goats using the molecular method, as well as investigating the morphological and the ultrastructural characteristics of the isolated species. A total of 1000 esophagi were collected from sheep and goats and examined for the presence of sarcocysts. Macroscopic sarcocysts were isolated from the infected esophagi, and Sarcosystis species were identified molecularly by 18S rRNA gene sequence analysis. Moreover, the ultrastructure of the sarcocysts was investigated by both scanning and transmission electron microscopy. The macroscopic sarcocysts were detected in 9.1% (91/1000) of the esophagi. The results of electron microscopy indicated the characteristic features of the macroscopic sarcocysts. The cysts contained numerous merozoites and banana-shaped bradyzoites. The bradyzoites were characterized by possessing a double-membrane pellicle and consisted of a conoid in one of the apices, numerous micronemes, two rhoptries, as well as a long, convoluted mitochondrion, subterminal nucleus, and several amylopectin granules. The partial analysis of the 18S rRNA gene presented that all isolates produced bands of expected sizes on gel electrophoresis. The findings from the phylogenetic analysis revealed that the identified Sarcocystis species were most closely related to S. gigantea, S. moulei, and S. medusiformis. To the authors’ knowledge, this is the first time S. medusiformis has been recorded in goats. Goats and sheep can be proposed as alternative intermediate hosts for S. gigantea and S. moulei, respectively, cross-infection may also occur between them and the host specificity of these species of Sarcocystis is questionable.

This study aimed to record the prevalence of different Sarcocystis spp., demonstrate their morphological characters by laboratory, histopathological and immunohistochemical techniques, also to investigate the pathological lesions in esophagus, heart, and brain samples collected from naturally infected donkeys in Egypt. The current study identified four Sarcocystis spp. (S. bertrami, S. equicanis, S. asinus and S. fayeri) in meat samples and S. neurona in the brain of the examined donkeys. Examination of meat samples showed macroscopic tissue cysts of S. bertrami andmicroscopic tissue cysts of S. equicanis/S. asinus and S. fayeri. According to the thickness of the cyst wall, Sarcocystis spp. were divided into smooth thin walled S. bertrami, ciliated thin walled S. equicanis/S. asius and striated thick walled S. fayeri. Also, immunohistochemical (IHC) examination showed strong labeled schizonts and cyst wall of S. neurona, while the bradyzoites appeared negative staining. Pathological examination revealed degeneration of muscles with intermuscular edema and few mononuclear inflammatory cells, but brain tissue showed perivascular lymphocytic cuffing and satellitosis with developing schizonts. The current study concluded that the accurate recognition about Sarcocystis spp. in donkeys would be helpful in their treatment, prevention and control measures.

Objective Sarcocystosis is an important zoonotic protozoal disease with worldwide distribution and wide range of hosts. The aim of the present study was to determine the intensity of Sarcocystis spp. infection and to show histopathological features of their cystic lesions in slaughtered cattle of Zabol- Iran. Methods From April to September 2018, 500 tissue samples from esophagus, heart, diaphragm, tongue and masticatory muscles were prepared from 100 slaughtered cattle. All samples were fixed in 10% neutral buffered formalin and routine tissue processing protocol was performed. Results The microscopic results showed that 92.2% of specimens had thin-walled cysts of S. cruzi and 14% had thick-walled Sarcocystis (S. hirsuta and S. hominis) but macrocyst was only observed in one cattle. The positivity rate of thin walled cysts was 58.8% for heart, 13.9% for masticatory muscles, 10.2% for tongue, 9.3% for esophagus and 7.8% for diaphragm. The positivity rate of thick walled cysts was 32.8% for esophagus, 28.6% for tongue, 22.9% for heart, 15.7% for masticatory muscles and 0% for diaphragm, which could represent either S. hominis or S. hirsuta. The most infected tissue was heart and the least infected tissue was diaphragm. Thin walled cysts (S. cruzi) were mostly found in heart and were less found in diaphragm. However, thick-walled cysts (S. hirsuta and S. hominis) were mostly detected in esophagus. No thick-walled cysts were found in diaphragm muscle. Conclusion A high positivity rate of sarcocystosis in slaughtered cattle in Zabol abattoir revealed heavily environmental contamination of Sistan region by this important parasitic disease.

Samples of diaphragm were collected from 53 sika deer from Gifu Prefecture, Japan; 220 sarcocysts were isolated, examined in wet mounts and classified according to their cyst wall protrusions. The sarcocysts were then examined molecularly in order to assign them to different species. All but 11 of the 220 sarcocysts were initially identified by means of a multiplex PCR assay targeting cox1 of five species, whereas the remaining 11 sarcocysts were identified by standard PCR and sequencing. DNA from selected sarcocysts was used for PCR amplification and sequencing of cox1 (59 sequences) and 18S rDNA (23 sequences). The 220 sarcocysts comprised seven major cox1 sequence types or species. Types 4 and 7 were assigned to the known species Sarcocystis pilosa and Sarcocystis ovalis, whereas types 1, 3 and 5 were considered to represent three new species, for which the names Sarcocystis japonica, Sarcocystis matsuoae and Sarcocystis gjerdei have been proposed. Types 2 and 6 were most similar to Sarcocystis tarandi and Sarcocystis taeniata, respectively, but could not be unequivocally assigned to these species. Sarcocysts belonging to S. japonica were macroscopic with fairly thick finger-like protrusions, whereas most sarcocysts of the six other species were microscopic. Sarcocysts of S. cf. tarandi and S. matsuoae were spindle-shaped and possessed thin finger-like cyst-wall protrusions. Sarcocysts of S. pilosa and S. gjerdei had similar hair-like protrusions, whereas those of S. cf. taeniata had a smooth surface. Sarcocysts of S. japonica, S. pilosa, S. cf. tarandi, S. gjerdei, S. matsuoae, S. cf. taeniata and S. ovalis were found in 50 (94.3%), 29 (54.7%), 22 (41.5%), 10 (18.9%), 8 (15.1%), 6 (11.3%) and 1 (1.9%) of the 53 sika deer examined, respectively. An improved multiplex PCR assay targeting cox1 was developed, through which the seven Sarcocystis spp. found in the present study could be identified. Keywords: Sarcocystis japonica, Sarcocystis matsuoae, Sarcocystis gjerdei, Cervus nippon centralis, 18S ribosomal RNA gene, Cytochrome c oxidase subunit I gene, Japan

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.

BackgroundBirds are one of the groups involved in the development of Sarcocystis Lankester, 1882, serving either as intermediate or definitive hosts. The white-tailed sea eagle Haliaeetus albicilla (Linnaeus, 1758), red kite Milvus milvus (Linnaeus, 1758) (both Accipitriformes) and common starlings Sturnus vulgaris Linnaeus, 1758 (Passeriformes) were examined to elucidate their participation in the development of Sarcocystis, as well as to determine the specific identity of the parasites based on morphological and especially molecular analyses.MethodsIn 2020–2021, one white-tailed eagle, one red kite and five common starlings were parasitologically examined for the presence of Sarcocystis using flotation centrifugation coprological method and by wet mounts of intestinal mucosa scrapings and/or muscle samples. Positive samples were processed by light microscopy, histologically and followed molecularly at four genetic markers (18S rRNA, 28S rRNA, ITS1 and cox1).ResultsThe white-tailed eagle harboured oocysts/sporocysts of S. arctica Gjerde et Schulze, 2014 in the intestinal mucosa, while the intestinal mucosa of the red kite and breasts and leg muscles of one common starling were positive to S. halieti Gjerde, Vikøren et Hamnes, 2018. Sequences from eagle shared 99.6 − 100% identity with each other and S. arctica in the red fox (V. vulpes Linnaeus, 1758) from the Czech Republic. Sequences from the common starling and red kite shared 100% identity with each other and with S. halieti in the great cormorant (P. carbo [Linnaeus, 1758]) from Lithuania and H. albicilla from Norway.ConclusionsThe white-tailed sea eagle (H. albicilla) acts as natural definitive host of S. arctica, whereas the common starling (St. vulgaris) and red kite (M. milvus) represent intermediate and definitive hosts, respectively, for S. halieti.

Barbary sheep (Ammotragus lervia) is a North African native wild Caprinae, introduced in the 1970s in new territories such as Spain, the USA, and Mexico. Here, we describe Sarcocystis species in Barbary sheep. Sarcocysts were found in 19 out of 56 adult A. lervia in Southern Spain and characterized morphologically and molecularly. By light microscopy, sarcocysts had thin ( 2 μm) walls. By transmission electron microscopy, sarcocysts with thick walls had Type 14 villar protrusions corresponding to S. tenella/S. capracanis of domestic sheep (Ovis aries) or goats (Capra hircus). Sarcocysts with thin walls had Type 7b villar protrusions that corresponded to S. arieticanis/S. hircicanis of domestic sheep or goats. Molecular analyses allowed the identification of only thick-walled Sarcocystis species. Six sarcocysts were assigned to S. tenella (99.2–100% and 95.6–100% sequence similarity within 18S rRNA and COI, respectively) and 19 sarcocysts were assigned to S. capracanis (98.5–99.8% and 97.9–99.0% sequence similarity within 18S rRNA and COI, respectively). Further studies are needed for taxonomic identification of sarcocysts in Barbary sheep because Sarcocystis species in sheep and goats are not cross transmissible despite morphological similarities.

Background Sarcocystis species are obligatorily heteroxenous parasites, of which some are zoonotic, representing a public health and economic impact. This study investigated the occurrence of Sarcocystis spp. in cattle sampled from a Belgian slaughterhouse. Methods A total of 200 carcasses were included in the study, sampled during 10 sampling days. The sedimentation method was applied to isolate the sarcocysts from both heart and diaphragm muscles collected from each carcass. Multiplex PCR, PCR–RFLP as well as cox1 gene sequencing techniques were applied serially on collected sarcocysts for species identification. Results Sarcocystis spp. were detected in 64% (128/200; 95% CI 57–71%) of the sampled carcasses. Female dairy cattle presented the highest Sarcocystis occurrence rate (91%) as well as the highest Sarcocystis species diversity compared to female beef and male beef. Sarcocystis spp. were detected more often in the heart muscles than in the diaphragm among female beef (p p = 0.001), while in male carcasses no significant difference was observed (p = 0.763). The effect of age was not significant in male carcasses (p = 0.872), while the odds of finding sarcocysts significantly increased with age (p = 0.003) within both types of female carcasses. S. cruzi was the most prevalent species and was found in 56.5% (113/200) of the carcasses, followed by S. hominis (21.0%, 42/200), S. bovifelis (12.5%, 25/200), S. bovini (2.0%, 4/200), S. hirsuta (1.5%, 3/200) and S. heydorni (0.5%, 1/200). Six different species were detected in the diaphragm, while only two species were recovered from the heart. S. cruzi was the most prevalent species in heart, while in the diaphragm, this was S. hominis. Conclusions The detection of S. hominis in 21% of the sampled carcasses presents a potential food safety issue, and further research is warranted into controlling this infection. Graphical Abstract

Protozoan parasites of the genus Sarcocystis are obligatory heteroxenous cyst-forming coccidia that infect a wide variety of animals and encompass approximately 200 described species. At least four Sarcocystis spp. (S. falcatula, S. neurona, S. lindsayi and S. speeri) use opossums (Didelphis spp.) as definitive hosts, and two of them, S. neurona and S. falcatula, are known to cause disease in horses and birds, respectively. Opossums are restricted to the Americas, but their distribution in the Americas is heterogeneous. Five Didelphis spp. are distributed in South America (D. aurita, D. albiventris, D. marsupialis, D. imperfecta and D. pernigra) whereas just one opossum species (D. virginiana) is found in North America. Studies conducted in the last decades show that Sarcocystis spp., derived from South American Didelphis spp., have biological and genetic differences in relation to Sarcocystis spp. shed by the North American opossum D. virginiana. The aim of this review was to address the peculiar scenario of Sarcocystis species shed by South American opossums, with a special focus on diagnosis, epidemiology, and animal infections, as well as the genetic characteristics of these parasites., Parasitos protozoários do gênero Sarcocystis são coccídios heteroxenos formadores de cistos, que infectam variadas espécies animais e compreendem cerca de 200 espécies descritas. Pelo menos quatro Sarcocystis spp. (S. falcatula, S. neurona, S. lindsayi e S. speeri) utilizam gambás (Didelphis spp.) como hospedeiros definitivos; e duas delas, S. neurona and S. falcatula são conhecidas por causarem doença em equinos e aves, respectivamente. Gambás didelfídeos são restritos ao continente americano, contudo são distribuídos de forma heterogênea nas Américas. Cinco Didelphis spp. são distribuídos na América do Sul (D. aurita, D. albiventris, D. marsupialis, D. imperfecta e D. pernigra), enquanto somente uma espécie (D. virginiana) é encontrada na América do Norte. Trabalhos conduzidos, nas últimas décadas, mostram que Sarcocystis spp. derivados de Didelphis spp. sulamericanos possuem diferenças biológicas e genéticas, quando comparados a Sarcocystis spp. excretados pelo gambá norte-americano D. virginiana. O objetivo desta revisão é discutir a situação peculiar das espécies de Sarcocystis na América do Sul com um foco especial em diagnóstico, epidemiologia e infecções animais, assim como nas características genéticas desses parasitos., Facultad de Ciencias Veterinarias

Various muscle tissue samples from 60 moose (Alces alces) in the Baltic region were examined for Sarcocystis species. Sarcocysts were detected in 49 out of 60 (81.7%) moose investigated. Six species, Sarcocystis alces, Sarcocystis hjorti, Sarcocystis linearis, Sarcocystis silva, Sarcocystis ovalis, and Sarcocystis sp., were identified using light microscopy (LM), and DNA sequence analysis (cox1 and 18S rDNA). Sarcocysts of S. alces, S. ovalis, and S. hjorti were studied using transmission electron microscopy (TEM); sarcocyst walls of S. alces, S. ovalis, and S. hjorti were type 25, type 24, and type 7a, respectively. Sarcocystis linearis previously found in roe deer and red deer was also shown to use moose as an intermediate host for the first time. The unknown Sarcocystis sp. was rare and might employ another main intermediate host. Phylogenetic results demonstrated that Sarcocystis sp. was most closely related to Sarcocystis tarandivulpes, using canids as definitive hosts.

The parasites from buffaloes in Guizhou province, southwest China, were suspected to be sarcocysts through morphologic observation. 18S rRNA and the subunit I of cytochrome oxidase (cox1) sequences...

Background Sarcocystis species are intracellular protozoan parasites that can pose a threat to animal health and food safety. The aim of this study was to investigate the prevalence of infection with Sarcocystis infection in sheep from China. Results In total, 52.51% (335/638) of tissue samples from domestic sheep contained sarcocysts through examination by light microscopy. The organisms were identified as S. tenella and S. arieticanis by molecular assays. Macroscopic S. gigantea and S. medusiformis were not found. The average sarcocysts loading was 18.07 ± 29.87 per square centimeter in the myocardium of domestic sheep. Furthermore, two specimens of argali (Ovis ammon) were examined and sarcocysts were found in the myocardium of one animal. According to the sequence of the cox1 gene of sarcocysts from argali, it was speculated as S. tenella. Conclusions We found a high prevalence and parasite load of Sarcocystis in sheep from both central and northwest China. This report is the first to indicate that argali may be a natural intermediate host for S. tenella.

Canids and scavenger birds were shown to act as definitive hosts of numerous Sarcocystis species using members of the Cervidae family as an intermediate host, whereas definitive hosts spreading closely related S. elongata, S. entzerothi, S. japonica, S. matsuoae, S. rangiferi, S. truncata, S. silva and S. tarandi remain unknown. In the current study, the intestine samples of 40 American minks (Neovison vison) were molecularly tested for the presence of the above-mentioned Sarcocystis spp. Species-specific PCR of cytochrome c oxidase subunit I (cox1) fragments and subsequent sequencing revealed the presence of sporocysts/oocysts of five species, S. elongata (n=2), S. entzerothi (n=10), S. japonica (n=4), S. silva (n=13) and S. truncata (n=21) in the analysed samples. Sarcocystis infection was confirmed in 32/40 (80%) examined samples. In addition, half of the infected animals (50%) were infected with multiple Sarcocystis species suggesting that American minks had access to meat of different deer species, such as roe deer, red deer and sika deer. This causes concern about compliance of hunters and game processing companies with game waste management rules. Further research on the involvement of mustelids in the transmission of various Sarcocystis spp. from different geographical locations is needed.

Parasites of the genus Sarcocystis infect birds, reptiles and mammals. They are characterised by an obligatory two-host life cycle. Sarcocystis spp. form sarcocysts in the muscles of intermediate hosts, once the host becomes infected through food or water contaminated with oocyst/sporocyst. Sporulating oocysts develop in the small intestine of definitive hosts when the animal consumes cyst-infected meat (Dubey et al. 2016)., Parasites of the genus Sarcocystis infect birds, reptiles and mammals. They are characterised by an obligatory two-host life cycle. Sarcocystis spp. form sarcocysts in the muscles of intermediate hosts, once the host becomes infected through food or water contaminated with oocyst/sporocyst. Sporulating oocysts develop in the small intestine of definitive hosts when the animal consumes cyst-infected meat (Dubey et al. 2016)

Sarcocystosis is a disease caused by various Sarcocystis species, a coccidian protozoan parasite that infects humans and animals and is commonly found in ruminants. Although Sarcocystis occurs all over the world, the species responsible for infecting buffaloes in Brazil have not been identified. In this study, we used molecular methods to estimate the prevalence of natural Sarcocystis infection in buffaloes. Phylogenetic analyses were conducted for the first time to identify the species of this protozoan that are responsible for infecting buffalos in southern Brazil, Rio Grande do Sul state. Heart samples from 80 buffaloes were subjected to microscopic examination followed by molecular analysis. Microcysts were present in 19 (23.75%) of 80 samples. The genomic DNA from the 19 cyst samples was extracted and amplified, and six sequences were obtained. The analysis was performed with the StandenPackage software, and the nucleotide sequences generated were analyzed by comparison with sequences in GenBank. All the sequenced samples were identified as Sarcocystis levinei. HIGHLIGHTS

The diaphragm muscles of 77 free-ranging red deer (Cervus elaphus) were examined for Sarcocystis species in Lithuania. Sarcocysts were detected in 61 out of 77 (79.2%) animals investigated. A total of 60 isolated sarcocysts were identified to species using subunit I of cytochrome c oxidase (cox1) sequence analysis. Overall, seven species, S. entzerothi, S. hjorti, S. iberica, S. linearis, S. pilosa, S. truncata and S. venatoria, were confirmed in Lithuanian red deer. Sarcocystis entzerothi was reported in red deer for the first time. Previously this species was shown to use sika deer as well as roe deer and fallow deer as an intermediate host. Based on cox1, with the addition of the current data, altogether 13 Sarcocystis species have so far been shown to use red deer as an intermediate host. Species detected in red deer demonstrated considerable differences in intraspecific genetic variation at cox1. Genetic distances between different samples of S. hjorti and S. linearis were calculated using principal coordinates analysis (PCoA), implying molecular divergence of same Sarcocystis species using different hosts in the same geographical area and divergence of those employing same intermediate host species from different areas.

This study aimed to determine the prevalence and molecular characterization of Sarcocystis spp. in bovine minced meat that is sold in various grocery stores and butcher shops in Van Province of Turkey. For this purpose a total of 150 samples were obtained from İpekyolu, Tuşba, Edremit, Erciş, and Gevaş districts of Van Province in monthly periods from May to October 2019. 28% (42/150) were found positive for Sarcocystis species as a result of the microscopic analyses and 96.6% (145/150) were found positive for Sarcocystis species as a result of the multiplex-PCR and RFLP methods. Sarcocystis cruzi (96.6%) was detected in all samples that were detected positive using molecular methods. Sarcocystis hominis-like was found in 143 (95%) samples whereas S. hirsuta was detected in only 4 (2.6%) samples. According to the Basic Local Alignment Search Tool (BLASTN) analysis of the 18S rRNA gene region of the S. cruzi (MN832695) and S. hirsuta (MN832697) isolates, they showed 100% similarity to the samples (MH681972; MH681973) that were submitted to GenBank from China. The BLASTN analysis of the 18S rRNA gene region of the S. hominis-like isolate (MN832696) revealed that it was 99.45% identical to the S. bovini (KT901155) isolated from a water buffalo in New Zealand. In conclusion, the molecular characterization of Sarcocystis spp. has been provided for the first time in Van Province, and the first unverified scientific data for S. bovini has been established in this study.

Having examined 19 great cormorants (Phalacrocorax carbo) hunted in Lithuania, sarcocysts were found in the muscles of two birds. Sarcocysts detected were examined using light microscopy (LM), transmission electron microscopy (TEM), and 18S rDNA, 28S rDNA, ITS1, cox1, and rpoB sequence comparison. Based on the molecular analysis, mainly of the ITS1 region, sarcocysts were identified as Sarcocystis halieti. This is the first Sarcocystis species characterised in the great cormorant. Under the LM sarcocysts were ribbon-shaped, very long and thin (the largest fragment found amounted to 6.5 × 0.1 mm) with a smooth and thin (up to 1.2 μm) cyst wall. Banana-shaped bradyzoites were 7.2 × 1.9 (6.3-8.2 × 1.4-2.4) μm. Under TEM, the cyst wall was wavy, 0.8- to 1.2-μm thick. The comparison of 12 species demonstrated cox1 and rpoB to be unsuitable for the identification of Sarcocystis spp. using birds as intermediate hosts.

The presence of Sarcocystis cysts in the muscle tissue of domestic dogs (Canis familiaris), which normally serve as definitive hosts, is unusual and infrequent. Here, S. caninum sarcocysts were identified for the first time in two of 37 dogs (2.7%) from China. Examination using light microscopy found that the S. caninum sarcocysts were up to 1520 μm long and up to 147 μm wide and contained numerous 1.5-3.3 μm wedge-like villar protrusions (vp). Transmission electron microscopy revealed that the sarcocysts had pleomorphic vp that closely resembled those of "type 9c." Five loci, 18S rDNA, 28S rDNA, mitochondrial cox1, ITS1 and ropB, were sequenced and characterized in S. caninum sarcocysts. The sequences of the five loci shared similarities of 99.9-100%, 99.0-100%, 99.4-100%, 99.6-100%, and 99.7-100%, respectively, with those of S. arctica. Phylogenetic analysis based on the sequences of 28S rDNA and mitochondrial cox1 indicated that S. caninum and S. arctica are closely related to Sarcocystis species that use a raptorial bird as their definitive host.

Small parts of esophagus and cardiac muscles of abattoir sheep from Qena, Upper Egypt between June 2016 to May 2017 were examined by both light microscope (LM), transmission electron microscope (TEM), and DNA sequence analysis of 18 S rRNA gene to determine the rate of infection with Sarcocystis spp. and identify the parasite according to morphological, ultrastructure characteristics, and DNA sequence analysis. LM and TEM studies revealed that microscopic, thin walled sarcocysts were detected in 47 of 63 male (74.6 %) and in 32 of 41 female (65.07 %) as a spindle-shaped with size (225 – 431.3 μm x 34.7- 82.4 μm) in esophagus muscles, and ovoid with size (141.6- 263 μm x 51.4- 82.3 μm) in cardiac muscles. The result sequences were compared with other previously sequenced Sarcocystis species retrieved from Gene Bank it was most closely refered to Sarcocystis tenella (identity 99- 100%).

This study was conducted to determine theinfection rate of Sarcocystis species in sheep in Assiut. A total of 100 slaughtered sheep examined for Sarcocystis, no macroscopic cysts were observed, 93% (93/100) were positive by microscopical examination and 28% (28/100) were positive by using Agar gel diffusion test.Theinfection rate of Sarcocystis had very high significant differences between microscopic examination and Agar gel diffusion test. High statistical significant effect was found on the infection rate of Sarcocystis in different examined muscles of sheep. The highest infection rate was recorded in oesophagus (71%), followed by diaphragm (65%), tongue (58%), skeletal muscles (53%) and heart muscles (43%). The infection rate of Sarcocystis in males was 95.2% (40/42) while in females were 91.4% (53/58) by microscopical examination. Higher infection rate of Sarcocystis were detected in sheep 6 months - 2 years age 95% (38/40) than that of sheep between 2-4 years old 92.7% (38/41) and those equal or older than 4 years 89.5% (17/19). No significant difference between infection and age groups or sex of animals was observed. The use of transmission electron microscope (TEM) allowed the identification of S. tenella and S. arieticanis in sheep in Assiut. Examination of S. tenella cyst wall with TEM showed palisade-like villar protrusions while, examination of S. arieticanis cyst wall showed hair-like villar protrusions parallel to cyst wall.

Cattle are intermediate hosts for several Sarcocystis species with different definitive hosts. The present study, to our knowledge, is the first to determine the prevalence of Sarcocystis infection and morphologically and molecularly identify Sarcocystis species in cattle in Qena Governorate, Upper Egypt. The samples were collected from the heart and oesophagus muscles of 84 slaughtered cattle (76 males and 8 females) aged between 11 months and 3 years from slaughterhouses in different localities in Qena Governorate, Upper Egypt. The samples were macroscopically and histologically examined, and the molecular identification of the species was performed using 18S ribosomal subunit DNA through PCR and DNA sequencing. Infection was detected in 72 out of 84 animals (85.7%) and was more prevalent in males (76.2%) than in females (9.5). Using light microscopy, the microscopic sarcocysts were observed to be thin-walled. Sequencing and genotyping revealed one isolate that had 99 and 100% identity, respectively, to Sarcocystis cruzi, while another isolate had 95 and 99% identity to Sarcocystis hjorti. The present study is the first to identify Sarcocystis infection in cattle in Qena Governorate, Upper Egypt both morphologically and molecularly. Sarcocystis cruzi and S. hjorti species were isolated from cattle, which is of veterinary importance and indicates that morphologically similar Sarcocystis species are genetically distinct. Additionally, the results show that Sarcocystis species are not host-specific.

Domestic and wild pigs, as intermediate hosts, can harbor tissue cysts of three Sarcocystis species namely S. miescheriana, S. suihominis and S. porcifelis. Out of them, S. suihominis is zoonotic. Romania is a country with high consumption of raw and/or undercooked traditional pork products. This fact may greatly favor the acquiring of the zoonotic Sarcocystis infections by humans, as definitive host. Based on this consideration and in order to investigate the occurrence and public health significance of Sarcocystis spp. in two western counties (Caras-Severin and Timis) of Romania, a total of 165 heart samples from hunted wild boars (Sus scrofa, n=101) and home slaughtered domestic pigs (Sus scrofa domesticus, n=64) were screened using microscopic fresh examination and molecular methods. Microscopic examination revealed the presence of sarcocysts in 60.4% of wild boars, and 23.4% of domestic pigs. Genetic characterization of isolates through the PCR-RFLP procedure, targeting the 18S rRNA gene, was successfully achieved for all microscopically positive samples, indicating the presence of a single species, S. miescheriana, in both hosts. The identity of 13 selected S. miescheriana isolates was also confirmed through sequencing. The tested hosts older than 27 months were found to be significantly higher infected (p

Sheep is considered as a biological guide for the environmental pollution with different parasites because of their grazing habits and rearing patterns. Herein, we examined 540 sheep of different ages and sexes slaughtered at 3 Egyptian Provinces (Dakahlia, Damietta and Cairo), for detection of Sarcocystis infecion. Esophageal muscle specimens were collected from slaughtered sheep. Samples were examined using both visual inspection and muscle compression technique. Fast 5 minutes 10% Geimsa solution immersion of the fresh samples was perfect for better observation of the microscopic sarcocysts. Formalin-preserved positive samples were subjected to histopathological examination. Overall, 95.37% of the examined sheep were harboring sarcocysts. Two types of sarcocysts were revealed, macroscopic (1% of older sheep) and microscopic (84.43% in smaller and 95.37% in older sheep). The infection was proportionally increased with age, while females were had a higher prevalence than males. Incidences equal to or over than 90% were reported allover the year seasons. Morphologically, macroscopiccystswere ranged from 0.8 – 1 cm in length with thin and smooth walls, while the microscopic ones were ranged from 600 – 700 µm in length with thick cyst wall either striated or smooth. Detailed morphological features were given. This report illustrates the wide dispersal of the infective stages of Sarcocystis species (sporocysts) in the environment, and could be beneficial for further molecular studies.

Simple Summary In this article, we investigated the occurrence of Sarcocystis species in samples of sheep and goats obtained from slaughterhouses in Riyadh, Saudi Arabia. We searched for tongue, heart, esophagus, diaphragm and skeletal muscle tissues. Fragments of these tissues were investigated by macroscopic evaluations, direct optical microscopy of tissue fragments, optical microscopy of digested fragment sediment, transmission electron microscopy and PCR followed by nucleotide sequencing. From the set of information obtained, we searched 230 sheep, and 84 goats; 91 and 36 were found to be infected, respectively. Transmission electron microscopy (TEM) revealed Sarcocystis tenella (S. tenella) in sheep and Sarcocystis capracanis (S. capracanis) in goats. Sarcocystis species were confirmed in Saudi Arabian sheep and goats by molecular testing. S. capracanis was most closely related to S. tenella, with the mitochondrial cytochrome c oxidase subunit I gene (COX1) sequences sharing a (91.7%) identity. Abstract Sarcocystosis is induced by species of Sarcocystis, which is an intracellular protozoan parasite in the phylum Apicomplexa. The diversity and importance of Sarcocystis species in sheep and goats in Saudi Arabia are poorly understood. In this study, the tongue, esophagus, heart, diaphragm, and skeletal muscles were collected from 230 sheep and 84 goats, and the tissues were examined for the presence of Sarcocystis species by macroscopic examination and light microscopy. Microscopic Sarcocystis species cysts were found in both sheep and goats. Transmission electron microscopy (TEM) revealed S. tenella in sheep and S. capracanis in goats. Sarcocystis species were confirmed for the first time in Saudi Arabian sheep and goats by molecular testing. S. capracanis was most closely related to S. tenella, with the COX1 sequences sharing 91.7% identity. A phylogenetic analysis produced similar results and indicated that the Sarcocystis isolates were within a group of Sarcocystis species in which dogs were the final host. Finally, the Sarcocystis species cysts from sheep and goats could be grouped together, indicating that they were strongly related.

Rodents have been widely studied as intermediate hosts of Sarcocystis; however, only a few reports on these parasites in the black rat (Rattus rattus) are known. Having examined 13 black rats captured in Latvia, sarcocysts were found in skeletal muscles of two mammals and were described as Sarcocystis ratti n. sp. Under a light microscope, sarcocysts were ribbon-shaped, 0.9–1.3 × 0.09–0.14 mm in size and had a thin (0.8–1.3 μm) and smooth cyst wall. The lancet-shaped bradyzoites were 8.3 × 4.3 (7.5–9.3 × 3.9–4.8) μm. Under a transmission electron microscope, the cyst wall was up to 1.3 μm thick, wavy, the ground substance appeared smooth, type 1a-like. Morphologically, sarcocysts of S. ratti were somewhat similar to those of S. cymruensis, S. rodentifelis, and S. dispersa-like previously identified in the brown rat (Rattus norvegicus). On the basis of 18S rDNA, 28S rDNA, and cox1, significant genetic differences (at least 2.3, 4.5, and 5.8%, respectively) were observed when comparing S. ratti with other Sarcocystis species using rodents as intermediate hosts. While ITS1 sequences of S. ratti were highly distinct from other Sarcocystis species available in GenBank. Phylogenetic and ecological data suggest that predatory mammals living near households are definitive hosts of S. ratti.

Sarcocystis species are the obligate tissue protozoan parasites of livestock causing clinical and subclinical disease resulting in downgrading of the meat and their products thereby leading to economic losses. The present study reveals the prevalence and distribution of sarcocystosis in water buffaloes (Bubalus bubalis) slaughtered at local abattoirs in A.P for a period of 1 year from June 2014 to May 2015. A total of 137 buffalo carcasses were screened grossly and microscopically organ wise viz., esophagus, tongue, heart, diaphragm and intercostal muscles. Out of 137 screened, 91 were infected with an overall prevalence of 66.42 %. Age wise analysis of 89 young male calves aged about 18–36 months old revealed 65.16 % (58/89) and 48 old she buffaloes (5–8 years) showed 68.75 % (33/48). The organ wise prevalence was highest in esophagus (51.82 %) followed by tongue (47.44 %), heart (29.92 %), diaphragmatic muscles (28.46 %) and intercostal muscles (18.24 %), respectively. Morphometric studies revealed the presence of two Sarcocystis species, i.e., S.levinei and S.fusiformis infection with a prevalence of 43.79 and 22.62 %, respectively, along with mixed infection rate of 43.06 %. Microscopic studies of S.levinei showed sarcocyst length/width/cyst wall thickness ranged between 0.31–0.69/0.09–0.12 mm/

A great problem was evoked about the imported Indian buffalo meats which were used to compensate the decrease in the meat industry in Egypt. This is due to the infection of the Indian meat with Sarcocystis species specially the prevalent macroscopic S. fusiformis. Therefore, we planned this work in order to update the prevalence and distribution patterns of S. fusiformis infecting the Egyptian water buffaloes, as well as studying their ultrastructure. Tissue specimens from esophagus, heart, tongue, diaphragm and throat muscles were recovered from 550 water buffaloes slaughtered at Mansoura abattoir in Dakahlia Province, Egypt, during the period between July 2009 and June 2010. S. fusiformis cysts were recovered from 58.72% of the examined animals. The infection rate was higher in aged buffaloes than young ones. The prevalence of infection was 100% in the esophagus, while no infection was detected in the heart. Ultrastructurally, the cyst wall was thin (1-3µm) exhibiting highly branched and anastomosed cauliflower-like villar protrusions.

The study was carried at Mirha Exports private limited at Mohali, Punjab, from December 2012 to May 2013 during which 1000 buffaloes of 2–3 years of age slaughtered for meat purposes were screened for the Sarcocystis species. Most of the animals examined at anti-mortem appeared normal and clinically healthy. However, at slaughtering, the gross and microscopic examination of representative samples of striated muscles collected from slaughtered buffaloes revealed 95.50 % prevalence of Sarcocystis species. Out of 1000 samples examined, 955 samples (95.55 %) were infected with microscopic cysts with size ranging from 19.1 to 95.9 × 10.2 to 68.9 μm cyst, while 6 samples (0.60 %) had mixed infection with macroscopic cyst with size ranging from 1270 to 19,000 × 700 to 7000 μm cyst of white or creamy colour and microscopic cysts. Among the muscles infected with cysts, highest number of cysts were recorded in tongue muscles (87.40 %) followed by oesophageal muscles (83.60 %), cardiac muscles (53 %) and diaphragmatic muscles (31.30 %).

Due to the lack of molecular research conducted, little is known about Sarcocystis species diversity in the fallow deer (Dama dama). Until now, Sarcocystis jorrini and Sarcocystis morae were described to form sarcocysts in the muscles of this host. In the present study diaphragm muscle samples of free-ranging fallow deer from Lithuania were investigated for Sarcocystis species. Sarcocysts were detected in 39 out of 48 (81.3%) fallow deer examined. Under a light microscope two types of sarcocysts having hair-like and finger-like protrusions were observed. Based on DNA sequence analysis of cox1 and 18S rDNA, two species, S. morae and Sarcocystis entzerothi were identified. In prior studies, the latter species was only detected in Lithuanian roe deer (Capreolus capreolus) and in sika deer (Cervus nippon). The haplotype network of S. morae sequences specified close relationships between haplotypes found in the same country. According to current knowledge, the fallow deer is characterised by low Sarcocystis species richness as compared with other cervid species from Europe.