Prevalence and distribution pattern of Sarcocystis spp. in slaughtered cattle from the Peruvian tropical Andes, Peru.

This study aimed to estimate the prevalence and distribution patterns of Sarcocystis spp. in cattle tissues in Chachapoyas province in the Peruvian tropical Andes. Additionally, the risk factors associated with the prevalence and the correlation of two diagnostic techniques (direct microscopy of squashed fresh muscle tissues and histopathology) were explored. The tongue, heart, esophagus, Latissimus dorsi muscle, and diaphragm of 210 animals slaughtered in the municipal slaughterhouse of Chachapoyas were evaluated by both techniques. Macroscopic sarcocysts were detected in 16.7% of tissues (CI 95% 11.7-21.7%). The total prevalence of Sarcocystis spp. was 96.2% (95% CI 93.6-98.8%) by direct light microscopy and 100% by histopathology. The highest Sarcocystis prevalence was detected in the esophagus. No significant statistical differences were found in the prevalence of Sarcocystis related to sex, age, or provenance. Both techniques demonstrated a very weak Kappa correlation (κ ≤ 0.24) in predicting the presence of the parasite in each of the five evaluated muscles. Direct microscopy can be implemented at slaughterhouses as a rapid screening test, but it is essential to confirm by histopathology the absence of the parasite in direct-microscopy-negative samples. It is also recommended that beef from the Peruvian Andes be thoroughly cooked for both human and animal consumption because of the zoonotic potential of some species of Sarcocystis.

Detection of Sarcocystis hominis, Sarcocystis bovifelis, Sarcocystis cruzi, Sarcocystis hirsuta and Sarcocystis sigmoideus sp. nov. in carcasses affected by bovine eosinophilic myositis.

Bovine eosinophilic myositis is an inflammatory myopathy characterized by multiple focal or diffuse grey to green patches leading to condemnation of affected carcasses. Although its etiology is still uncertain, there is evidence that Sarcocystis species may play a role in the development of eosinophilic myositis. The goal of the present study was to identify Sarcocystis spp. in intralesional and extralesional tissues of condemned cattle carcasses, in order to evaluate the possible role of different bovine Sarcocystis spp. in the etiology of bovine eosinophilic myositis. Muscle samples (n = 100) of 26 affected carcasses were collected in Northern Italy. One to five samples with lesions and two aliquots of tissue without lesions were collected from each carcass; lesions were grossly categorized in green focal lesions and green diffuse patches. Genomic DNA was extracted and analyzed by multiplex-PCR targeting different Sarcocystis spp. Unidentified species were characterized morphologically (light microscopy, histology), ultrastructurally (scanning and transmission electron microscopy) and on the molecular level (complete 18S rRNA gene and partial cox1 gene sequencing). A bovine eosinophilic myositis prevalence of 0.017% was visually assessed by routine carcass inspection between 2014 and 2019 in Italy (184/1,108,150 slaughtered cattle). Out of 26 carcasses, 25 revealed the presence of at least one Sarcocystis species (96.2%). The presence of Sarcocystis spp. DNA was significantly more frequent in intralesional than in extralesional samples. Considering the different species, Sarcocystis bovifelis and Sarcocystis hominis were significantly more frequent in intralesional (41.7% and 50%, respectively) than in extralesional samples (1.9% and 15.4%, respectively), while there was no significant difference between the presence of Sarcocystis cruzi and Sarcocystis hirsuta in intralesional (27.1% and 2.1%, respectively) and extralesional (30.8% and 1.9%, respectively) samples. The presence of an unnamed Sarcocystis sp. showing thick-walled (3.7-5.4 µm) cysts with densely packed, flattened, undulating and narrow protrusions, which showed an S-shape in side view, was recorded in the diaphragm of two carcasses. Genomic DNA from individual sarcocysts isolated from the diaphragm was successfully amplified and further sequenced. Sequence comparison revealed <94.6% and 83.4% identity at 18S rRNA and cox1 genes, respectively, with other named Sarcocystis spp., while the phylogenetic analysis clearly separated the unnamed Sarcocystis sp. from the other Sarcocystis spp. using cattle as intermediate hosts. The present study contributes to the understanding of the importance of different Sarcocystis spp. in the pathogenesis of bovine eosinophilic myositis. The results emphasize the association of Sarcocystis hominis and Sarcocystis bovifelis with bovine eosinophilic myositis and highlight the presence of a new Sarcocystis sp. using cattle as intermediate hosts. The name Sarcocystis sigmoideus sp. nov. is proposed for the newly described Sarcocystis species.

Sarcocystis in horses from Rio Grande do Sul, Brazil: Molecular identification of Sarcocystis bertrami and Sarcocystis neurona in muscle tissues.

Horses are intermediate hosts of Sarcocystis spp. capable of forming cysts in their musculature. This study aimed to detect sarcocysts and investigate the presence of nucleic acids from Sarcocystis spp. in samples of striated muscles from horses in the State of Rio Grande do Sul, Brazil, necropsied at the Veterinary Pathology Laboratory of the Federal University of Santa Maria. A total of 108 samples were collected from 24 horses and examined through direct examination. Microscopic tissue cysts were observed in three samples: tongue (2) and esophagus (1) from two animals. Extractions were performed on the found cysts and tissues, even though sarcocystosis detection was not present. DNA samples were subjected to Nested-PCR using Tg18s primers, and the amplified products were subjected to Restriction Fragment Length Polymorphism (RFLP) using DdeI and HpaII enzymes. DNA belonging to Sarcocystis spp. was amplified in tissues from 91.7% (22/24) of the equines, and 67.6% (73/108) of the samples tested positive in the Nested-PCR reaction. The tissues with the highest detection frequency were: diaphragm 92.3% (12/13), gluteal muscle 77.2% (17/22), and esophagus 66.7% (4/6). In RFLP, Sarcocystis spp. was detected in 21 tissues from 11/22 equines, and cysts, identified through nucleotide sequencing, were determined to be S. bertrami. S. neurona was identified in 11 samples from 7/22 animals, with co-infection detected in 5/22 cases. The high detection rate indicates a concerning circulation of the protozoan, particularly the zoonotic S. bertrami found in all tissues, which are commonly exported for human consumption.

Identification of Sarcocystis spp. in synanthropic (Muridae) and wild (Cricetidae) rodents from Argentina.

The occurrence of Sarcocystis species was investigated in synanthropic (Muridae) and wild (Cricetidae) rodents from Argentina. Nine species were captured (n = 356). Sarcocysts were detected in muscles of 8.7% (31/356) and 3.7% (4/106) of the rodents by histopathology and direct microscopic observation, respectively. PCR-sequencing targeting the 18S rRNA, cox1, and ITS1 regions was performed on samples with positive histopathology. Four different 18S rRNA sequences or sequence groups with high intra-group identities (99.6-100%) were detected in Mus musculus, Oxymycterus rufus, Akodon azarae, and Necromys lasiurus. Eight sequences showed 99.5-99.7% identity with S. dispersa. Thirteen sequences showed low identity (95.3-96.4%) with other Sarcocystis spp. The obtained coxI sequences (n = 9) were almost identical to each other and showed a high similarity with S. strixi (99.2-99.5%) and S. lutrae (99.1%), despite the 18S rRNA sequences from the same samples suggested the occurrence of at least two species. This suggests that coxI may not show high variability in Sarcocystis spp. that use rodents as intermediate hosts. Six ITS1 sequences were obtained, showing high identity but low coverage with several Sarcocystis spp. Multilocus sequence typing and BLAST analysis did not lead to an accurate species identification. Possible reasons are the detection of new species or the limited molecular information available from previously described Sarcocystis spp. Phylogeny suggests that the detected Sarcocystis spp. may use raptor birds or snakes as definitive hosts. This study represents the first molecular identification of Sarcocystis spp. in naturally infected rodents of the Cricetidae and Muridae families in South America.

A Systematic Meta-Analysis of Global Sarcocystis Infection in Sheep and Goats.

Sarcocystosis is an intracellular parasitic disease caused by Sarcocystis spp. that has a worldwide prevalence. Symptoms of the disease include diarrhea and muscle pain. The disease poses a threat to the health of animals. The aim of this review is to investigate the global prevalence of Sarcocystis infection in sheep and goats during 2013-2022. We searched five databases: Web of Science, Science Direct, PubMed, Scopus, and Google Scholar. A total of 36 articles containing 44 datasets met the criteria and were included in the study. The total infection rates of Sarcocystis in sheep and goats were 66.3% (95% CI, 51.79-79.38%) and 52.1% (95% CI, 29.45-74.23%), respectively. It was found that Sarcocystis species tend to have a host species preference. Coinfection of S. tenella and S. arieticanis often occurred in sheep, and goats were frequently infected with S. capracanis. Age and sex were identified as risk factors for Sarcocystis infection in sheep and goats. The infection rates of female and male animals were significantly different, with females having a higher infection rate. Age-adjusted analysis showed that infection rates in animals older than one year were higher than in animals younger than one year. This study unveiled the global distribution of Sarcocystis and sheds light on its transmission in sheep and goats.

First report of Sarcocystis spp. (Apicomplexa, Sarcocystidae) in Lagostomus maximus (Desmarest, 1917) (Rodentia, Chinchillidae) in Argentina.

Sarcocystis is a genus of intracellular parasitic protozoa that infects various species of mammals, birds, and reptiles worldwide. At least 46 Sarcocystis species naturally infect rodents as intermediate hosts producing tissue cysts. This study aimed to provide the first report and molecular characterisation of Sarcocystis spp. in muscles from plains viscacha (Lagostomus maximus) in Argentina. Muscle samples of 53 plains viscachas from three provinces of Argentina were processed by homogenisation and optical microscopy to detect tissue cysts. Positive samples were analysed by PCR-sequencing, using the following markers: 18S rRNA, ITS1, and coxI. The 18S rRNA and coxI consensus sequences were aligned with other sequences from Sarcocystis spp., and phylogenetic trees were constructed. Of all animals processed, 13.2% (7/53) harboured Sarcocystis sp. cysts. 18S rRNA consensus sequences were obtained from four muscle samples and one individual cyst, and they showed 99.88-100% similarity, except for the cyst sequence, which showed 97.11% homology. Similarities of only 96-97% were recorded in the 18S rRNA fragment with other Sarcocystis spp. whose sequences are available in the GenBank. The five coxI fragment sequences obtained were 100% identical and showed an identity of 99.41-99.48% with S. canis. For ITS1 only short and low-quality sequences were obtained. In the phylogenetic trees, all the sequences from plains viscachas were positioned together in a branch separated from other Sarcocystis spp. These results could be related to new Sarcocystis spp. producing sarcocysts in plains viscachas. Besides, comprehensive cyst morphological analysis using TEM from the new Sarcocystis species will allow a description of the cyst wall ultrastructure. In this sense, further studies are needed to deepen these findings and elucidate other potential intermediate and possible definitive hosts.

Sarcocyst Quantification and Viability: Freezing Treatment as an Alternative to Carcass Condemnation.

PURPOSE: The inspection of animal products is important for controlling parasitic zoonoses. Some processes that guarantee food safety to consumers such as carcass condemnation cause economic losses. This study aimed to detect Sarcocystis cysts in cattle hearts obtained from slaughterhouses and to evaluate sarcocyst viability after freezing treatment. METHODS: When myocardial tissues were minced and subjected to fresh examination, sarcocysts were observed in all analyzed tissues resulting in 21.73 cysts/g of tissue. Sarcocyst viability was verified after tissue freezing at 35 ± 2 °C and - 20 ± 2 °C for 0-12 h. After freezing, the tissues were minced, and sarcocysts were collected and stained with Tripan Blue. In addition, cysts were mechanically disrupted to check bradyzoite viability. RESULTS: Cysts and bradyzoites were unviable at - 35 °C for ≥ 3 h and - 20 °C for ≥ 8 h. CONCLUSION: These results suggest freezing treatment as an alternative to condemnation of cattle carcasses contaminated with Sarcocystis spp. Similar studies using freezing treatment with other animals infected by Sarcocystis must be conducted.

Sarcocystosis among buffaloes from slaughterhouses in Nile Delta, Egypt: morphologic assessment and molecular confirmation.

Background: Sarcocystis species are coccidian protozoan zoonotic parasites of the phylum Apicomplexa. There is a large diversity of Sarcocystis species. Some of them are pathogenic and dangerous to humans, domestic, and wild animals. Cattle are common intermediate hosts. The infection of meat with different species of Sarcocystis can be serious for public health. Aims: The current study aimed to determine the prevalence of sarcocystosis in slaughtered buffaloes in Tanta city abattoirs, Nile Delta, Egypt. Methods: Morphological and histological examinations and a molecular study were undertaken. A total of 517 locally bred buffaloes were slaughtered in Tanta city, Egypt. Each buffalo carcass was visually inspected for the presence of Sarcocystis macrocysts. Fifty tissue samples containing suspected cysts were examined by using different techniques including histology, transmission electron microscope (TEM), and PCR. Results: By visual inspection, the overall prevalence of suspected sarcocystosis was 26.5%. The highest infection rate was detected visually from the esophagus followed by skeletal muscles and diaphragm whereas the least was recorded in the tongue. Histological and TEM examination showed that the cysts were packed with bradyzoites separated by multiple septa. 100% of the sarcocysts diagnosed visually in the esophagus and muscles were confirmed by PCR, compared to only 25% of those detected in the tongue. Conclusion: These results highlight the high prevalence of sarcocystosis among buffaloes in Egypt, possibly due to widespread environmental contamination by Sarcocystis oocysts. The use of molecular methods should be encouraged to confirm the identity of the suspected cysts.

Molecular detection and identification of three intracellular parasites of retail mutton products in Beijing, China.

Sarcocystis spp., Neospora caninum and Toxoplasma gondii are globally ubiquitous pathogens, and domestic sheep are considered to be one of the intermediate hosts. 83 myocardial samples of sheep were collected from 12 retail stores in Beijing, China. Sarcocystis spp., N. caninum and T. gondii were identified by PCR amplification of the 18S rRNA gene, Nc-5 gene and 529bp DNA fragment with a prevalence of 86.7% (95% CI: 77.5-93.2) and 43.4% (95% CI: 32.5-54.7) for Sarcocystis spp. and N. caninum infections, respectively, and no T. gondii was detected. The co-infection prevalence of Sarcocystis and N. caninum was 38.6% (95% CI: 28.1-49.9). Two Sarcocystis species were subtyped by analyzing 18SrRNA sequences and were identified as Sarcocystis tenella and Sarcocystis arieticanis. The prevalence of S. tenella and S. arieticanis infections was 84.3% (95% CI: 74.7-91.4) and 56.6% (95% CI: 45.3-67.5), respectively. This study shows that sheep have a high risk of infection with Sarcocystis and N. caninum, suggests that effective prevention measures are needed to avoid the spread of these parasites in sheep. Toxoplasmosis in sheep poses a threat to human and animal health and requires monitoring and preventing continuously.

Molecular Epidemiology, Species Distribution, and Zoonotic Importance of the Neglected Meat-Borne Pathogen Sarcocystis spp. in Cattle (Bos taurus): A Global Systematic Review and Meta-analysis.

BACKGROUND: Sarcocystis species are diverse apicomplexan parasites, though only two zoonotic species (S. hominis and S. heydorni) circulate between cattle and humans. Due to the importance of cattle in the human food chain and to prevent the consequences of parasitosis in humans, the first global systematic review and meta-analysis on molecular epidemiology, species distribution, and zoonotic significance of Sarcocystis infection in cattle was performed. METHODS: For this aim, four international English databases (PubMed, Scopus, Google Scholar, and Web of Science) were systematically searched till 20th September 2021, and random-effect models were drawn to calculate total estimates and their 95% confidence intervals (CIs). RESULTS: Finally, 44 papers from 21 countries were qualified for this review which examined 8526 cattle regarding Sarcocystis infection, rendering a total prevalence of 62.7% (95% CI 53-71.5%). Globally, 12 Sarcocystis spp. have been reported from cattle, including S. cruzi, S. hominis, S. hirsuta, S. rommeli, S. heydorni, S. bovifelis, S. bovini, S. sinensis, S. gigantea, S. fusiformis, S. hjorti and S. tenella. Among them, S. cruzi (37 studies), S. hominis (22 studies) and S. hirsuta (19 studies) were the 3 most common species, with 76.4% (95% CI 64.8-85%), 30.2% (95% CI 19.3-44%) and 8.7% (95% CI 3.8-18.6%), respectively. However, molecular identification was not performed in 48.4% (95% CI 27.3-70.1%) of the positive samples. CONCLUSION: Despite the zoonotic significance of Sarcocystis spp., particularly S. hominis, the epidemiology and distribution of Sarcocystis infection in cattle remains unclear and demands more extensive researches around the world.

Detection and phylogenetic analysis of Sarcocystis moulei and Sarcocystis spp. (Sarcocystidae: Apicomplexa) from slaughtered sheep in southwest Iran.

Sarcocystis species are intracellular protozoan which mostly complete their life cycle in two hosts. The parasite has a significant economic, medical and veterinary impact in many regions of the world and considered as a significant health problem in Iran. However, most of infections are asymptomatic and mortality is extremely rare. The present study aimed to determine the molecular phylogeny of the Sarcocystis species isolated from sheep slaughtered in southwest Iran, using mitochondrial DNA sequences of 18 S rRNA gene. The DNA was extracted from sheep muscular tissue (n = 60), and partial sequence of 18 S rDNA was amplified and sequenced. Phylogenetic analysis revealed two monophyletic clades representing S. moulei (n = 3) and Sarcocystis spp. (n = 3). BI posterior probability and MP bootstrap values strongly supported the monophyly of these clades. In conclusion, phylogenetic analysis of Sarcocystis species using 18 S rRNA gene could be helpful in identifying the new species of the Sarcocystis.

Prevalence and molecular assessment of Sarcocystis infection in livestock in northeast Iran.

Sarcocystis is an intracellular parasite of the apicomplexa phylum. More than one hundred species of Sarcocystis can infect wildlife and livestock animals. Samples of the liver, heart, muscle and diaphragm were collected from sheep, goats and cattle in Gonabad, northeast Iran and subjected to macroscopic, microscopic, tissue digestion, sequencing and phylogenetic analyses of 18S-rRNA region. Tissue sections stained with hematoxylin and eosin were also provided for surveying sarcocysts in the samples. Tissue digestion showed that all animal samples (100%) were infected with Sarcocystis bradyzoites. Phylogenetic analysis indicated that out of 50% of sheep genotypes belonged to S. tenella, and 20% to S. moulei and S. arieticanis. Moreover, three samples of macroscopic specimens of sheep were identified as S. gigantea, 100% of cattle isolates infected with S. cruzi, 80% of goat isolates belonged to S. capracanis and one macroscopic specimen of goat (20%) identified as S. moulei. Sarcocystis infection is highly prevalent in livestock in northeast Iran, where a variety of Sarcocystis species are unequivocally circulating in the region.

Detection of Sarcocystis spp. and Toxoplasma gondii in swine and detection of DNA of these protozoa in tissues and sausages / Soroprevalência de Sarcocystis spp. e Toxoplasma gondii em suínos e detecção do DNA desses protozoários em tecidos e embutidos

Abstract The seroprevalence of Sarcocystis spp. and Toxoplasma gondii was researched in swine raised in Santa Maria, RS, Brazil. Serum samples from 84 pigs from 31 farms were tested using indirect immunofluorescence assay (IFA) for both agents. Additionally, 53 samples of pork sausages and tissues destined for human consumption, including: salami, sausage, black pudding, heart, tongue, brain, and rib muscle, were submitted to PCR to detect DNA for each agent. The frequency of anti-Sarcocystis spp. antibodies was 36.9% (31/84), with titers ranging from 32 to 1024, and 25% (21/84) for anti-T. gondii antibodies, with titers ranging from 64 to 2048. Sarcocystis spp. and T. gondii DNA were detected in 67.9% (36/53) and 13.2% (7/53) of samples, respectively. The presence of antibodies and the detection of DNA from Sarcocystis spp., and T. gondii suggests that the pigs were infected and may serve as an important reservoir for both parasites. The infection by these protozoa in the swine population is relevant to public health due to their zoonotic potential.

Resumo A soroprevalência de Sarcocystis spp. e Toxoplasma gondii foi pesquisada em suínos criados em Santa Maria, RS, Brasil. Amostras de soro de 84 suínos de 31 fazendas foram testadas pela reação deimunofluorescência indireta (IFA) para ambos os agentes. Adicionalmente, 53 amostras de embutidos suínos e tecidos cárneos destinados ao consumo humano, incluindo: salame, linguiça, morcela, coração, língua, cérebro e músculo da costela foram submetidas à PCR para detecção de DNA para cada agente. A frequência de anticorpos anti-Sarcocystis spp. foi de 36,9% (31/84), com títulos variando de 32 a 1.024; e 25% (21/84) para anticorpos anti-T. gondii, com títulos variando de 64 a 2048. A presença de DNA de Sarcocystis spp. e T. gondii foi detectada em 67,9% (36/53) e 13,2% (7/53) das amostras avaliadas, respectivamente. A detecção de anticorpos e DNA de Sarcocystis spp. e T. gondii sugere que os suínos foram infectados e podem servir como um importante reservatório de ambos os parasitas. A circulação desses agentes na população suína é relevante para a saúde pública devido ao seu potencial zoonótico.

First Report of Sarcocystis Masoni in a Captive Alpaca (Vicugna Pacos) From China.

Background: Sarcocystosis is a parasitic disease caused by intracellular protozoan parasite of the genus Sarcocystis. Tissue samples of alpacas (n = 4) from Henan province (China) were screened for Sarcocystis spp. infection by histological examination, pepsin digestion, and molecular assays. Results: Sarcocystis spp. was detected in heart, liver, spleen, lung, and kidney of an alpaca by molecular assays. Many sarcocysts with inflammation responses were observed in this alpaca myocardium, and they showed a high similarity to Sarcocystis masoni by sequence analysis. Conclusion: This study is the first to demonstrate Sarcocystis spp. infection in alpaca from China. The higher parasite load in the alpaca myocardium indicated that it had contact with an environment contaminated with sporocysts, and that the alpaca was susceptible to Sarcocystis spp.

Sarcocystis species in bovine carcasses from a Belgian abattoir: a cross-sectional study.

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 < 0.001) and dairy carcasses (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.

Molecular detection of cattle Sarcocystis spp. in North-West Italy highlights their association with bovine eosinophilic myositis.

BACKGROUND: Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa because of the evidence supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based on either morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis. METHODS: To reach our aim, individual striated muscle samples from BEM condemned carcasses (N = 54) and diaphragm muscle samples from randomly sampled carcasses (N = 59) were obtained from Northwest Italy slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S rDNA and cox1 genes. PCR products amplified using the genus-specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis were sequenced to achieve species identification. RESULTS: Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species in slaughter cattle (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses (42.6% and 1.8%, respectively) and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in two carcasses. CONCLUSIONS: Our study contributes to updating the data on the prevalence of the different Sarcocystis spp. in cattle in Italy, highlighting the presence of three Sarcocystis spp., S. cruzi, S. hominis and S. bovifelis, in BEM lesions and allowing us to speculate on the possible role of S. hominis and S. bovifelis as the major sarcosporidian species involved in bovine eosinophilic myositis.

Sarcocystidae in wild birds of southeastern Brazil / Sarcocystidae em aves silvestres do sudeste do Brasil

Abstract This study aimed to identify members of the Sarcocystidae family in naturally infected wild birds at a rescue center in the state of Minas Gerais, southeastern Brazil. The heart and brain of 44 wild birds were evaluated by bioassay in mice to detect T. gondii, and extracted DNA was used for nested PCR of the 18S ribosomal DNA gene to detect members of the Sarcocystidae family. The positive samples were sequenced, assembled, edited and compared with sequences deposited in GenBank. Toxoplasma gondii was isolated from six (13.6%) out of 44 birds. Toxoplasma gondii DNA was identified in 10/44 (22.7%) of the birds. The amplified sequences exhibited 100% similarity with the DNA of the ME49 strain of T. gondii. Sarcocystis DNA (99% similarity) was identified in 5/44 (11.4%) of the birds. T. gondii and Sarcocystis spp. are common in wild birds in Minas Gerais, Brazil.

Resumo O objetivo deste estudo foi identificar membros da família Sarcocystidae em aves silvestres de vida livre naturalmente infectadas e resgatadas no estado de Minas Gerais, Brasil. Coração e cérebro de 44 aves silvestres foram avaliados por bioensaio em camundongos para detecção de T. gondii e extração de DNA para Nested-PCR do gene 18S do DNA ribossomal de membros da família Sarcocystidae. As amostras positivas foram sequenciadas, analisadas, editadas e comparadas com sequências depositadas no GenBank. Toxoplasma gondii foi isolado de seis (13,6%) das 44 aves. DNA de T. gondii foi identificado em 10/44 (22,7%) das 44 aves. As sequências amplificadas exibiram 100% de similaridade com o DNA da cepa ME49 de T. gondii. DNA de Sarcocystis (99% de similaridade) foi identificado em 5/44 (11,4%) das 44 aves. T. gondii e Sarcocystis spp. são encontrados, comumente, em aves silvestres no estado de Minas Gerais, Brasil.

Epidemiological and Histopathological Investigation of Sarcocystis spp. in Slaughtered Dromedary Camels (Camelus dromedarius) in Egypt.

Sarcocystosis is considered one of the major parasitic diseases with a worldwide distribution. It is caused by the obligatory intracellular protozoan parasites of the genus Sarcocystis. Besides its public health issues, sarcocystosis results in significant economic losses due to its impact on productivity and milk yield. A wide range of final and intermediate hosts have been identified, including mammals, birds, and reptiles; however, few studies have investigated the contribution of camels to maintaining the epidemiological foci of the disease in countries such as Egypt. The present study was conducted to grossly and histopathologically identify the prevalence rate of Sarcocystis spp. in camels (N = 100) from the Aswan Governorate, Egypt. Furthermore, the major risk factors related to the development of sarcocystosis in camels were investigated. Samples from the diaphragm, cardiac muscle, esophagus, and testes of the slaughtered camels were collected. Interestingly, Sarcocystis was detected in 75% of the examined camels. Following the studied variable factors, camels aged 5 years or more were found to be at higher risk, with an infection rate of 87.7% (57 of 65) than those younger than 5 years. The infection rate was 81.4% (57 of 70) in males and 60% (18 of 30) in females. The esophagus was the most affected organ (49%), followed by the diaphragm (26%) and cardiac muscle (17%), whereas none of the testes samples were affected. Taken together, the present study demonstrates the high prevalence of Sarcocystis in the examined camels and suggests the importance of these animals in preserving the epidemiological foci of sarcocystosis in Egypt. Future research should map the circulating strains in Egypt and aim to raise public health awareness about the importance of sarcocystosis and other related zoonotic diseases.

Identification of Sarcocystis Spp. in One-humped Camels (Camelus dromedarius) from Riyadh and Dammam, Saudi Arabia, via Histological and Phylogenetic Approaches.

Sarcocystis (S.) spp. are intracellular protozoan parasites that infect birds and animals, resulting in substantial commercial losses. Sarcocystis spp. have an indirect life cycle; canines and felines are known to act as final hosts, and numerous domestic and wild animals act as intermediate hosts. The presence of sarcocysts in camel meat may diminish its commercial quality. There is limited knowledge regarding the taxonomy and diagnosis of Sarcocystis spp. that infect camels in Saudi Arabia. In this study, transmission electron microscopy (TEM) revealed S. cameli and S. camelicanis (camelicanis) in Camelus (C.) dromedarius. This is the first report of S. camelicanis in Saudi Arabia and is considered a significant finding. Based on cytochrome c oxidase subunit I gene (COX1) sequences, two samples of Sarcocystis spp. isolated from C. dromedarius in Riyadh and Dammam were grouped with S. levinei hosted by Bubalus bubalis in India, S. rangi hosted by Rangifer tarandus in Norway, S. miescheriana hosted by Sus scrofa in Italy and S. fayeri hosted by Equus caballus in Canada. The sequences obtained in this study have been deposited in GenBank.

Sarcocystis infection in beef and industrial raw beef burgers from butcheries and retail stores: A molecular microscopic study.

Sarcocystis is a genus of eucoccidian parasites, which globally infects humans and various animals. In addition to economic losses in livestock industries, the parasite is a zoonosis that infects humans through contaminated beef and pork with the parasite sarcocysts. Therefore, this study was carried out to assess Sarcocystis contamination in beef and industrial raw beef burger samples from butcheries and retail stores in Tehran, Iran. Overall, 180 samples of 90 beefs and 90 raw industrial beef burgers with at least 80% meat were randomly collected in Tehran, Iran. Samples were studied microscopically after peptic digestion. Furthermore, sample genomic DNAs were used in conventional polymerase chain reaction (PCR) to amplify approximately 900-bp fragments from 18S ribosomal DNA. Of 180 samples, 170 samples (94.4%) were microscopically and 161 samples (89.44%) were molecularly positive for Sarcocystis spp. Eucoccidial DNA fragments were detected in 161 samples (89.4%), including 78 (86.6%) beef and 83 (92.2%) beef burger samples. No significant differences were found between the beef and beef burger infestations by Sarcocystis bradyzoites using statistical analysis (P > 0.05). Statistically significant differences were seen between the sample type and the intensity of parasites in samples (P = 0.003). Furthermore, differences between the conventional PCR results (positive/negative) and the intensity of parasites in samples were statistically significant (P < 0.001). The considerable prevalence of Sarcocystis spp. in beef and beef burger samples reflects high transmission of the parasite in meat producing cattle, which is important due to food hygiene. Although the most prevalent bovine species, S. cruzi, is not a zoonosis, it is highly recommended to follow guidelines on the parasite transmission prevention due to the existence of S. hominis as a zoonotic bovine species.