Sarcocystis Infection Research Articles

Sarcocystosis in Ruminants of Iran, as Neglected Food-Borne Disease: A Systematic Review and Meta-analysis.

Sarcocystis is a zoonotic parasitic pathogen which endangers the safety of meat and meat products. This systematic review and meta-analysis aimed to evaluate the prevalence rate and status of Sarcocystis spp. in ruminants as important food sources in Iran. Data were collected from papers indexed in five English language electronic databases (PubMed, Scopus, Web of Science, Science Direct, and Google Scholar) and four Persian electronic databases (IranMedex, SID, IranDoc, and Magiran) from January to April 2019. Papers were selected based on inclusion criteria. Data analysis was performed in StatsDirect statistical software, version 2.7.2. The searching process resulted in the identification of 73 studies. Data analyses revealed that the total prevalence (95% confidence intervals) of Sarcocystis spp. in Iranian ruminants was 74.40% (64.01-83.56). In addition, a significant association was also observed between sarcocystosis infection in Iranian ruminants and year, host, location, and diagnostic technique (P < 0.001). According to our data, the prevalence of Sarcocystis infection in ruminants is relatively high. High pathogenicity of some Sarcocystis spp. and the negative impact that the spread of some parasites among ruminants can have on human and animal health necessitate the direction of more attention toward monitoring, controlling, and preventing sarcocystosis.

Prevalence and Microscopic Studies of Mixed Sarcocystis Infection in Naturally Infected Sheep and Goats in Soran City, Erbil- Iraq

Prevalence and Microscopic Studies of Mixed Sarcocystis Infection in Naturally Infected Sheep and Goats in Soran City, Erbil- Iraq

Muscular Sarcocystosis in Sheep Associated With Lymphoplasmacytic Myositis and Expression of Major Histocompatibility Complex Class I and II.

Sarcocystosis is a protozoal disease affecting a wide range of animals. The aims of this study were to characterize the following in sheep: (1) the muscle pathology in Sarcocystis infection, (2) the inflammatory infiltrate and its relationship to severity of infection, and (3) immune markers expressed by parasitized muscle fibers and parasitic cysts. Skeletal muscle samples from 78 sheep slaughtered in southern Italy were snap frozen and analyzed by histopathology, immunohistochemistry, and immunofluorescence. Polymerase chain reaction (PCR) and sequencing were used for Sarcocystis species identification. All 40 muscle samples tested were PCR-positive for Sarcocystis tenella. Histologically, cysts were identified in 76/78 cases (97%), associated with an endomysial infiltrate of lymphocytes and plasma cells. The T cells were predominantly CD8+, with fewer CD4+ or CD79α+ cells. Eosinophils were absent. Notably, sarcolemmal immunopositivity for major histocompatibility complex (MHC) I and II was found in 76/78 cases (97%) and 75/78 cases (96%), respectively, both in samples with and in those without evident inflammatory infiltrate. The number of cysts was positively correlated with inflammation. In addition, MHC I was detected in 55/78 cyst walls (72%), and occasionally co-localized with the membrane-associated protein dystrophin. The findings suggest that muscle fibers respond to the presence of cysts by expression of MHC I and II. The possible role of MHC I and II in the inflammatory response and on the cyst wall is also discussed.

Sarcocystis spp. in Romanian Slaughtered Cattle: Molecular Characterization and Epidemiological Significance of the Findings.

Species of the genus Sarcocystis are recognized as protozoan parasites infecting a wide range of animals, including humans. This study aimed to provide data on the occurrence, genetic characterization, and epidemiological significance of Sarcocystis spp. in cattle destined for human consumption in Romania. A total of 117 heart samples from slaughtered cattle in three southwestern Romanian counties (Dolj, Timiș, and Gorj) were analyzed in order to detect sarcocysts, using fresh examination microscopic techniques. Subsequently, the isolated sarcocysts and/or cyst fragments (5–15 per sample) from each infected animal were molecularly characterized. Overall, 17.9% (21/117) of the tested animals were found to be Sarcocystis spp. positive by microscopy. Genetic characterization of Sarcocystis spp. isolates, based on sequence analysis of the 18S rRNA gene, showed the presence of a single species, namely S. cruzi. No correlation was found (p > 0.05) between S. cruzi infection and the origin, age, breed, and gender of cattle, but the grazing farming system was positively associated (p=0.031) with the pathogen prevalence and can be considered a risk factor (OR = 3.6) in acquiring infection. To evaluate the possible public health risk, further investigation focused on the processing of other Sarcocystis-specific tissue matrices and evidence of human infections is recommended. This is the first study of bovine Sarcocystis infection in Romania.

Sarcocystis rileyi in UK free-living wildfowl (Anatidae): surveillance, histopathology and first molecular characterisation

BackgroundReports from UK hunters of ‘rice grains’ in muscles of shot wildfowl (Anatidae) coincided temporally with the finding of sarcocystosis in a number of ducks found as part of the...

Detection of Humoral and Cellular Immune Responses in Buffaloes Naturally Infected with Sarcocystosis with Risk Factor Assessment

AbstractSarcocystosis is a silent, parasitic disease which affects various animal species and causes significant economic losses. It is caused by a number of different intracellularSarcocystisspp. This study was aimed to detect the host humoral and cellular immune response due to natural infection. Adding to the determination of the infection rate in Monufia Governorate, Egypt. A total number of 127 Egyptian buffaloes (Bubalus bubalis); 30 males and 97 females between 2-11 years of age were examined during 2018. An infection rate of 74% (94/127) was detected by macroscopic examination. The old age females were found to be at a high risk of 90.7% (88/97) in comparison with the young males 20% (6/30). Immunologically, the cellular and humoral immune response was determined using ELISA. A marked down-regulation of the proinflammatory Th-1 cytokine (IFN-γ) and up-regulation of the anti-inflammatory Th-2 cytokine (IL-5) adding to a high level of IgG and IgE were detected in the infected animals compared to the non infected ones. The local cellular immune response in the infected tissues was characterized by an accumulation of mixed inflammatory cells, granuloma formation, eosinophilic infiltration, muscular edema, and necrotic degeneration. In conclusion, theSarcocystisinfection rate in the naturally infected buffaloes in Monufia Governorate was high. This is the first study to provide a fundamental insight into the immune profile in buffaloes infected withSarcocystisspp. So, it will provide valuable insights to develop novel effective vaccines in future studies. Moreover, sensitive and specific tools should be established for the accurate diagnosis of this disease in the different Egyptian governorates through well-structured serological surveys.

Muscular Sarcocystis infection in ruminants slaughtered at Municipality abattoir and selected Hotels in Hawassa city, southern Ethiopia: Prevalence and associated risk factors.

Sarcocystosis is a parasitic zoonosis caused by Sarcocystis spp. which are Apicomplexan parasites requiring intermediate and definitive hosts to complete their life cycle. Although the parasite has worldwide distribution in man and many species of animals, the prevalence in most parts of Ethiopia is not clearly known. This cross sectional study was conducted between Nov. 2016 and May 2017 to estimate the prevalence of Sarcocystis infection in ruminants slaughtered in the municipality abattoir and at selected hotels of Hawassa city and to assess the potential risk factors. Data were collected through gross and histopathological examination of myocardial and esophageal muscles sampled from a total of 561 ruminants (176 sheep, 181 goats and 204 cattle). The overall prevalence of Sarcocystis infection in ruminants was 68.98%. All of the cysts were microscopic, and found more frequently (p < .05) in heart (62.08%) than esophageal muscle (45.45%) although concurrent infection of both organs was observed in 33.87% of the ruminants examined. No significant association was noted between Sarcocystis infection and the origin, sex and species of ruminants examined (p > .05). The higher prevalence recorded in the study area can be explained by the abundance of stray dogs, cats and wild carnivores that are roaming in the villages, lack of proper latrine, uncontrolled disposal of condemned offals and carcass, and provision of uncooked meat for dogs and cats. Thus, farmers' awareness creation and strategies targeted at breaking the life cycle of the parasite are required to reduce the prevalence of the parasite and thereby the foreseen zoonotic and economic impact.

Molecular identification targeting cox1 and 18S genes confirms the high prevalence of Sarcocystis spp. in cattle in the Netherlands

The reported prevalence of Sarcocystis infection in cattle in Europe ranges between 66 and 94%. Although in the Netherlands a prevalence of 100% was reported in 1993, this study aimed to develop a method for sensitive and specific molecular detection and species identification of Sarcocystis spp., in order to provide more recent data on the prevalence and identification of these protozoa in cattle meat intended for human consumption in the Netherlands. For this purpose, 104 cattle samples were obtained from Dutch slaughterhouses. Genomic DNA was extracted, and analysed by 18S and cox1 PCR. Magnetic capture was used to extract and amplify 18S-specific DNA. Sarcocystis DNA was detected in 82.7% of the samples. PCR amplicons of both targets were sequenced, and sequence identities of ≥97% were observed for Sarcocystis cruzi (65.4%), Sarcocystis hominis (12.5%), Sarcocystis bovifelis (8.7%), Sarcocystis hirsuta and Sarcocystis heydorni (both 1.0%). Mixed infections were observed in 17.3% of the samples. The magnetic capture was not significantly more sensitive compared with standard DNA extraction, but magnetic capture did add to the overall sensitivity. Using cox1 sequencing, all species are clearly distinguished, whereas for 18S the variation between species is limited, which particularly hampers reliable identification of thick walled Sarcocystis spp. Furthermore, the detection of 12.5% S. hominis and 1% S. heydorni points towards an established transmission route between cattle and humans in the Netherlands. The availability of four additional well-identified and well-referenced S. hominis cox1 sequences in public databases enables development of species-specific diagnostic PCRs targeting cox1, which in combination with magnetic capture could provide the means to determine the prevalence of human sarcocystosis.

Investigation of Parasitic Sarcocystis Infection in Native Poultry Carcasses in North Part of Iran, Mazandaran (Amol)

Investigation of Parasitic Sarcocystis Infection in Native Poultry Carcasses in North Part of Iran, Mazandaran (Amol)

Molecular screening approach to identify protozoan and trichostrongylid parasites infecting one-humped camels (Camelus dromedarius)

Little is known about the diversity of many parasites infecting camels, with most relying on morphological parameters. DNA extracted from different tissues (n = 90) and fecal samples (n = 101) from dromedary camels (Camelus dromedarius) in Egypt were screened for multiple parasites using different molecular markers. Screening of tissue samples (heart) for Toxoplasma gondii and Sarcocystis spp. was performed using B1 and 18S rRNA gene markers, respectively. T. gondii was further genotyped using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP). Sarcocystis was analyzed using PCR-RFLP characterization (XbaI and MboI restriction enzymes). A taxonomically challenging but important group of nematodes (Trichostrongylidae family) were screened using the ITS-2 ribosomal DNA (rDNA) species-specific markers. Furthermore, nested PCR was used for the detection of Cryptosporidium spp. (SSU rRNA gene) and positive samples were genotyped after RFLP (SspI and VspI) and sequencing. Cryptosporidium parvum isolates were subtyped by sequence analysis of the 60-kDa glycoprotein gene. This study revealed that many parasites infect the investigated camels, including T. gondii (1.1%), Sarcocystis spp. (64.4%), Cryptosporidium spp. (5.9%) and Trichostrongylidae nematodes (22.7%). The species contribution for nematodes was as follows: Haemonchus spp. (95.6%), Trichostrongylus axei (26%), Trichostrongylus colubriformis (65.2%) and Cooperia oncophora (60.8%). Mn-PCR-RFLP typing for Toxoplasma was only successful for three markers: 5′-SAG2 (type II), 3′-SAG2 (type II) and alt. SAG2 (type II). PCR-RFLP using XbaI showed possible mixed Sarcocystis infection. Moreover, the Cryptosporidium genotypes detected were C. parvum (IIdA19G1 and IIaA15G1R1), Cryptosporidium rat genotype IV and a novel genotype (camel genotype). This approach revealed the unique Cryptosporidium genotypes infecting the investigated camels, and the high genetic diversity of the investigated parasites.

Molecular differentiation of five Sarcocystis species in sika deer (Cervus nippon centralis) in Japan based on mitochondrial cytochrome c oxidase subunit I gene (cox1) sequences.

Several surveys of Sarcocystis infection in sika deer in Japan have shown a high prevalence, but the identification has been unclear because molecular data have been lacking or have been limited to 18S ribosomal RNA gene sequences. Thus, in our previous study based on such sequences, some Sarcocystis isolates from sika deer were not clearly separated from other species in the phylogenetic analysis. In the present study, we therefore characterized sarcocyst isolates from sika deer (Cervus nippon centralis) at the mitochondrial cytochrome c oxidase subunit I gene (cox1). Moreover, we developed a multiplex PCR based on cox1 sequences of all species found, so that we could rapidly identify sarcocysts of these species. Twenty-one sarcocysts from nine sika deer were examined. Five distinct cox1 sequence types, each with a high sequence identity (> 99%), were found, and the sarcocysts could thus be classified into five species. Based on the sequence comparisons and the phylogeny, Sarcocystis spp. of types 1, 3, and 5 are considered to represent three new species, which were most closely related to Sarcocystis silva/Sarcocystis truncata, Sarcocystis entzerothi, and Sarcocystis iberica/Sarcocystis venatoria, respectively. There was a slight uncertainly whether Sarcocystis sp. with type 2 sequences represented a new species or was identical to Sarcocystis tarandi. Type 4 sequences showed 99% identity with those of Sarcocystis pilosa from sika deer in Lithuania and have therefore been assigned to this species. In the multiplex PCR, type-specific fragments were successfully amplified for all five Sarcocystis spp., indicating that this assay may be useful for a rapid identification of sarcocysts of these species.

Prevalence and molecular characterisation of Sarcocystis miescheriana and Sarcocystis suihominis in wild boars (Sus scrofa) in Italy.

A sample of the diaphragm was collected from each of 100 wild boars legally hunted in the Val Grande National Park in north-western Italy and examined for the presence of Sarcocystis infection by histological and molecular methods. In histological sections, thick-walled sarcocysts consistent with those of Sarcocystis miescheriana were detected in 32 wild boars. Genomic DNA extracted from diaphragm samples was initially subjected to PCR amplification of the internal transcribed spacer 1 (ITS1) region, and 97 wild boars were found to harbour a Sarcocystis infection at this screening. Selected DNA samples were then subjected to PCR amplification and sequencing of the ITS1 region and the 18S and 28S ribosomal RNA (rRNA) genes of the nuclear ribosomal DNA unit, while all positive samples were subjected to PCR amplification of the mitochondrial cytochrome c oxidase subunit I (cox1) gene. S. miescheriana was identified in 97 wild boars (97%), while the zoonotic Sarcocystis suihominis was identified in one wild boar (1%), which also harboured S. miescheriana. Intra-specific sequence variation was found in all four DNA regions of S. miescheriana examined and in the 18S rRNA gene and ITS1 region of S. suihominis. The partial cox1 gene was amplified and sequenced from 72 isolates of S. miescheriana, yielding 43 haplotypes with pairwise sequence identities of 97.6-99.9%. These haplotypes were 79.1-79.8% identical with the cox1 sequence of S. suihominis. Phylogeny based on cox1 sequences placed S. miescheriana and S. suihominis as sister species within a clade comprising mainly Sarcocystis spp. of ruminants with felids as known or presumed definitive hosts. The same was true for the phylogeny based on 18S rRNA gene sequences.

Poultry carcasses investigation of parasitic sarcocystis infection in native in Mazandaran (North part of Iran)

Poultry carcasses investigation of parasitic sarcocystis infection in native in Mazandaran (North part of Iran)

Prevalence and molecular characterization of Sarcocystis infections of retail beef products from central China

Cattle are the intermediate hosts for five Sarcocystis species including S. hominis and S. heydorni, which also infect humans. To investigate the prevalence of Sarcocystis infections in beef products from 17 cities in the Henan Province of central China, 62 raw beef samples from markets were collected and analyzed for Sarcocystis presence via muscle squashing microscopic observation, histological section examination, and molecular characterization with 18S rRNA gene sequencing. Sarcocystis were detected in a total of 20 of the meat samples. Four species were identified that comprised S. cruzi, S. rommeli, S. heydorni, and S. hirsuta, with S. cruzi as the dominant species. In addition, seven of the 20 infected samples were infected with two or three species. Analysis of the 18S rRNA sequences recovered from these samples suggested very little genetic diversity within each species. This study represents the first molecular identification of Sarcocystis species infection in retail beef products from China. These findings will provide valuable information for evaluating the potential public health risk of bovine Sarcocystis species infections and the control of sarcocystosis in cattle.

Sarcocystis species in wild and domestic sheep (Ovis ammon and Ovis aries) from China

BackgroundSarcocystis 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.ResultsIn 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.ConclusionsWe 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.

Quality of beef diaphragm meat in naturally occurring Sarcocystic infection in cattle

The aim of this study was to investigate the possible harmful effect of Sarcocystis parasites on bovine diaphragm meat quality. Meat samples were collected from 120 bulls aged 20–24 months. Meat quality was investigated using microbiological and physico-chemical (RP-HPLC, GC) methods 48 hours after slaughter. Sarcocystis infection was associated with increased fat content, lightness L* and drip loss, and decreased ash and protein percentages. Infection also had a significant effect on the amount of amino acids (AAs), which slowly decreased as the number of sarcocysts increased. The total amount of AAs correlated with glutamic acid content (R = 0.966, P &amp;lt; 0.05). Heavily infected samples contained significantly lower amounts of putrescine, histamine, spermine and spermidine (P &amp;lt; 0.05) and&lt;br /&gt; a noticeable increase in the total count of aerobic microorganisms, but no change in the numbers of E. coli and coliform bacteria in comparison with no infected samples. Sarcocysts in beef diaphragms did not cause serious changes in the technological quality of the meat, but the biological quality of infected meat was reduced.

High prevalence of Sarcocystis spp. infections in cattle (Bos taurus) from central China

Myocardium and diaphragm samples of cattle (n = 521) from HeNan Province (China) were screened for Sarcocystis sarcocysts by histological examination, pepsin digestion, and molecular assays. Morphology and molecular assays were used for identification. The prevalence of Sarcocystis infection in cattle was 41.5% (216/521). Histological examination identified sarcocysts in the myocardium (49.4%, 200/405) and diaphragm (13.8%, 16/116) of cattle. Two species were identified, namely S. cruzi (41.3%, 215/521) and S. hominis (0.2%, 1/521). The findings of the present study indicate a high prevalence of S. cruzi infection in cattle from central China.

Sarcocystosis with thin Wall Cyst in Cardiac Muscle of Bullock : A Case Report

Sarcocystosis is a parasitic zoonoses caused by species of Sarcocystis, an intracellular protozoan parasite in the phylum Apicomplexa. In cattle Sarcocystis is mainly caused by Sarcocystis cruzi, Sarcocystis hominis and Sarcocystis hirsute. Bovine sarcocystosis caused by Sarcocystis cruzi mainly by ingesting the protozoan eggs through feed. The infection is mainly characterized by cyst formation in muscular tissue. Heart, diaphragm and skeletal muscles are the preferred locations for Sarcocystis spp in intermediate host and are more prevalent in adult animals (Rao and Hafeez, 2002). Bovine Sarcocystis results in weight loss, anorexia, decreased milk production, anemia, and prostration and in fatal cases often culminating in death of animals (Dafedar et al., 2011). The significance of bovine Sarcocystis infection prompted us to put the present case on record.

First molecular characterization and morphological aspects of Sarcocystis fusiformis infecting water buffalo Bubalus bubalis in Egypt.

Fresh muscle samples from water buffalo (Bubalus bubalis) aged 2-15, from Giza Province, Egypt; were examined for Sarcocystis infection. Macroscopic ovoid sarcocysts embedded in the muscle tissues of the examined buffaloes were detected; they measured 152-230 (210 ± 7) μm in length and 37-119 (95 ± 3) μm in width. The esophagus was the most infected organ followed by the diaphragm, and tongue, while the heart muscles were the least infected. The cyst cavity was compartmentalized by septa derived from the ground substance located under the primary cyst wall. Using transmission electron microscopy, the primary cyst wall bordered sarcocysts were determined to be 0.08-0.22 μm in thickness, raised from the parasitophorous vacuolar membrane, and surrounded by a secondary cyst wall of host origin. The primary cyst wall had irregular wall folds with numerous cauliflower-like projections of variable sizes and shapes accompanied by knob-like electron-dense elevations. 18S rRNA gene expression studies confirmed that the present parasite isolates belonged to the genus Sarcocystis. The sequence data showed significant identities (>90%) with archived gene sequences from many Eimeriidae organisms, and a dendogram showing the phylogenetic relationship was constructed. The most closely related species was Sarcocystis fusiformis KR186117, with an identity percentage of 98%. The recovered sequences were deposited in the GenBank under the accession number MG572125. The present study, to our knowledge, is the first collective ultrastructural and molecular study that confirmed the taxonomy of sarcocysts isolated from water buffaloes in Egypt as Sarcocystis fusiformis.

Detection and Identification of Sarcocystis cruzi (Protozoa: Apicomplexa) by Molecular and Ultrastructural Studies in Naturally Infected Korean Cattle (Bos taurus coreanae) from Daejeon, Korea

To survey the prevalence of Sarcocystis infections, 210 heart samples were collected from Korean native cattle (Bos taurus coreanae) at an abattoir in Daejeon Metropolitan City, Republic of Korea. Sarcocysts were detected form 31 specimens (14.8%) and identified as Sarcocystis cruzi via transmission electron microscopy. The wall of S. cruzi has flattened protrusions that did not contain fibrils or microfilaments. The protrusions arose irregularly from the base, contained a fine granular substance, lacked internal microfilaments, and measured approximately 0.21–1.25 μm in length and 0.05–0.07 μm in width. Sequence analysis revealed 99.5% homology to S. cruzi. This is the first report on the prevalence of S. cruzi in native cattle from the Republic of Korea.