Sarcocystis Gigantea Research Articles

Development of a highly specific LAMP assay for detection of Sarcocystis tenella and Sarcocystis gigantea in sheep.

Sarcocystis infection in sheep has caused significant economic losses in the livestock industry, and the genetic similarity among Sarcocystis species highlights the need for precise diagnostic methods in sheep. This study developed a loop-mediated isothermal amplification (LAMP) method targeting COX-1 and 28S rRNA genes to detect Sarcocystis tenella and Sarcocystis gigantea, respectively. The LAMP method exhibited high specificity, selectively amplifying target DNA sequences without cross-reactivity with closely related protozoa, such as Toxoplasma gondii and Neospora caninum. Detection limits were determined as 3 × 105 copies/L for S. tenella and 6 × 104 copies/L for S. gigantea, enabling sensitive identification of low-level infections. Comparative analysis with conventional PCR on sheep cardiac tissues demonstrated a higher LAMP detection rate (80.0% vs 66.7%). In conclusion, the LAMP method offers superior sensitivity to conventional PCR, allows visual confirmation of results, and provides a rapid diagnostic tool for identifying S. tenella and S. gigantea infection in sheep. However, due to the limitation of sample availability, we were unable to assess all Sarcocystis species that use sheep as intermediate hosts, which warrants further research.

Sentinels in the shadows: Exploring Toxoplasma gondii and other Sarcocystidae parasites in synanthropic rodents and their public health implications

Synanthropic rodents play a crucial role in maintaining the life cycle of Toxoplasma gondii in anthropized regions and can serve as indicators of environmental oocyst contamination. This investigation aimed to explore the occurrence of T. gondii infection within synanthropic rodent populations using a molecular diagnostic technique targeting the 18S rDNA gene, which is generic for Coccidia, with subsequent specific PCR confirmation. We examined 97 brown rats (Rattus norvegicus), 67 black rats (R. rattus), 47 house mice (Mus musculus), and 1 common shrew (Sorex araneus). PCR tests were conducted on the brain, heart, and tongue tissues. PCR tested positive in at least one of the examined tissues in 26 R. norvegicus (26.8%), 13 R. rattus (19.4%), and 13 M. musculus (27.6%). Sequencing comparisons by BLAST allowed us to identify four different species of cyst-forming Apicomplexa. In particular, T. gondii DNA was detected in 13 (6.1%) rodents, Hammondia hammondi (including H. hammondi-like organisms) in 36 (17%) subjects, Besnoitia sp. (in two cases identified as B. besnoiti) in 8 (3.7%), and Sarcocystis gigantea in two (0.94%). Rodents from peri-urban and urban environments can act as indicators of environmental contamination by oocysts of apicomplexan parasites with cats as definitive hosts, such as T. gondii, H. hammondi, and S. gigantea, the latter of which has never been previously recorded in rodents. Moreover, the presence of B. besnoiti, a parasite with an unidentified definitive host in Europe, sheds light on the potential role of these hosts as infection sentinels.

The histopathological and molecular diagnosis of myocardial sarcocystosis in sheep, and the immunohistochemical correlation of parasitic lesions with MMP - 2 and MMP - 9

This study was aimed at the histopathological and immunohistochemical level investigate cardiac tissue lesions caused by Sarcocystis spp. in sheep, and thereby, to collect detailed information on the presence of these parasites and the pathogenesis of sarcocystosis in ovine animals. PCR, a molecular diagnostic method, was used for the identification of the parasite species. Microscopic examination revealed that out of the 45 ovine cardiac tissue samples examined 23 contained Sarcocystis spp. cysts. The PCR analysis results showed that out of the 23 cardiac tissue specimens infected with sarcocysts, 7 were determined to be infected with Sarcocystis gigantea and 16 with Sarcocystis tenella. Immunohistochemical examination demonstrated that, semiquantitatively, MMP - 9 staining was stronger than MMP - 2 staining in the periphery of the parasite cysts.

Molecular Survey on Sarcocystis Species in Slaughtered Sheep in Hamedan, Iran

Introduction: Sarcocystis is an apicomplexan heteroxenous protozoan leading to adverse consequences for production in sheep with remarkable importance in public health. The current study aimed to investigate molecular prevalence data on Sarcocystis spp. in slaughtered sheep using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in Hamedan, Iran.
 Methods and materials: The carcass of 60 sheep slaughtered in the Hamedan abattoir were sampled from May 2019 to June 2020. Heart, scapular, esophageal and diaphragmatic muscles were macroscopically examined and recorded as positive in case of the presence of tissue cyst. In this regard, 50 grams of each muscle was sliced and transferred to the laboratory on ice for microscopic and molecular analysis. The molecular identification of Sarcocystis spp. was performed using the PCR-RFLP method.
 Results: By microscopy, all specimens contained bradyzoites in cysts. The molecular analysis further revealed Sarcocystis species, including Sarcocystis gigantea (S. gigantea), S. tenella, and S. arieticanis.
 Conclusion: The present study emphasized that three Sarcocystis spp. were circulating among sheep and carnivorous hosts in the investigated area. Further molecular investigations are highly recommended to better evaluate the epidemiology of this zoonotic parasite.

In Vitro Genotoxic Effects of Sarcocystis gigantea Cystizoites Acetone Powder Extract on Sheep Lymphocytes

The toxin of the protozoan intracellular parasite of sheep Sarcocystis gigantea is associated with many clinical and pathological signs. The aim of the study was to investigate In Vitro various chromosomal aberrations due to sarcocystosis infection. Macrocysts of Sarcocystis gigantea were isolated from local karadi sheep, homogenized with glass Dounce homogenizer; acetone powder was prepared from it and used in various concentrations to investigate the chromosomal aberration in vitro against sheep lymphocytes. The direct effects of parasite cystizoites acetone powder revealed various genotoxicity effects. These effects included chromosomal aberration (Isogap, Breaks and Dicentrics) and chromatids aberration (Gap and Deletion). It had also an effect on the mitotic index of the lymphocyte cells division. These genotoxicities were studied for the first time with in vitro technique using sheep lymphocytes. These results reflected that Sarcocystis gigantean parasite could cause structural and internal aberration in the chromosomes of their hosts.

Infection survey and morphological characteristics of Sarcocystis spp. in naturally infected Tibetan sheep from Qinghai in northwestern China.

Sarcocystosis is a parasitic disease caused by intracellular coccidian protozoans that belong to the genus Sarcocystis. These parasites can cause diseases of the nervous system, abortion and economically significant losses in host animals. Previous studies have reported that Sarcocystis is found in mammals, birds and reptiles, while molecular and morphological studies of infected Tibetan sheep have not been performed in the Qinghai region. The aim of this study was to determine the prevalence of Sarcocystis spp. in Tibetan sheep in Qinghai, northwestern China. The results showed that in 1155 samples, sarcocysts from unspecified species were found in 50% (577/1155) of the sheep tissues by microscopy detection. The positive rates of sarcocysts in the diaphragmatic, esophageal and cardiac muscles were 78.4% (175/223), 29.1% (207/711), and 88.2% (195/221), respectively. Ultrastructural features were exclusively observed in Sarcocystis gigantea in the esophageal tissues. The specific architecture was characterized as a space between the two layers of the original capsule wall, which was filled with fiber bundles and tissue fluid. Cauliflower-like protrusions of the original capsule wall were observed toward the outer surface of the capsule. Prominent protrusions contained fibers and matrix. In addition, the Sarcocystis 18S rRNA genes from 6 esophageal tissue samples were cloned, sequenced, and aligned to related sequences from GenBank. All 5S. gigantea sequences examined in this study were grouped into the same cluster and belonged to the same genotype. The other 5Sarcocystis tenella sequences were obtained from cardiac muscle and diaphragm muscle and belonged to the same clade. Overall, this study revealed a high infection rate of Sarcocystis in Tibetan sheep in the region. The results of this study may provide a reference for further research investigating the sarcocystosis epidemic in Qinghai, China.

A Heavy Infection Macro-cysts of <i>Sarcocystis</i> Spp. Case in a Najdi Sheep at Buraidah Slaughter House, Qassim, Saudi Arabia

Background: Protozoal diseases caused by <i>Sarcocystis</i> species can cause serious damage in sheep and goats flocks, inducing decreased growth conversion rates and partial or complete condemnation of carcasses at the slaughter house. Sarcocystis spp. affecting sheep and goats are host specific for their intermediate hosts and family specific for their final hosts. They derive their name from the intramuscular cyst stage present in the intermediate host. Objective: Macro and micro identification of macro-cysts of <i>Sarcocystis</i> spp. affecting a Najdi sheep carcass. Method: Detection of macrosarcocystis during routine post mortem examination and identification the detected cyst by histopathological investigation. Result: The macroscopic sarcocysts were found cleared and scattered in almost all skeletal muscles including shoulder, chest and diaphragm as well as thigh. Both morphological and histopathological characteristics indicated that the detected cysts may be identified as <i>Sarcocystis gigantea</i>. Conclusion: It is a case study of macro-cysts of <i>Sarcocystis</i> spp. affecting local breed of Najdi sheep. It is considered a seldom detected case all over many years during meat inspection at Buraidah slaughter house. The possible control measures are those of simple hygiene. Farm dogs should not be housed in, or allowed access to, fodder stores nor should they be allowed to defecate in pens where livestock are housed. It is also important that they are not fed uncooked meat.

Molecular characterisation of five Sarcocystis species in domestic sheep (Ovis aries) from Spain

The major aim of the present study was to determine by molecular methods whether the wide and narrow types of macroscopic sarcocysts in Spanish sheep belonged to different species, that is, Sarcocystis gigantea and Sarcocystis medusiformis, respectively. Additionally, we wanted to identify and characterize molecularly the species forming microscopic sarcocysts and determine the phylogenetic placement of all species found. Portions of the oesophagus, diaphragm and hind legs containing macroscopic sarcocysts were collected from slaughtered culled ewes at an abattoir in the Province of Madrid, Central Spain, but both macroscopic and microscopic sarcocysts were isolated for molecular examination. Genomic DNA from 63 sarcocysts (21 macroscopic, 42 microscopic) were examined at the cytochrome c oxidase subunit I gene (cox1), while selected isolates of each species found were further examined at the 18S and 28S ribosomal RNA (rRNA) genes. The 63 sarcocysts comprised five cox1 sequence types, each corresponding to a particular sarcocyst type, and thus represented five Sarcocystis spp. The slender fusiform and thick macrocysts belonged to S. medusiformis and S. gigantea, respectively. The microscopic sarcocysts belonged to Sarcocystis arieticanis, Sarcocystis tenella and a Sarcocystis mihoensis-like species with slanting thorn-like cyst wall protrusions, which was characterised molecularly for the first time. Based on its phylogenetic position, the S. mihoensis-like species probably uses corvids as definitive hosts.

Occurrence of Sarcocystis gigantea macrocysts and high frequency of S. tenella microcysts in sheep from southern Brazil.

The aim of this study was to investigate the occurrence of sarcocysts in sheep slaughtered in the Rio Grande do Sul state, Brazil. Heart and esophagus samples from 130 sheep were subjected to macroscopic and microscopic examination, followed by molecular analysis. Ten sheep (7.7%) had Sarcocystis gigantea macrocysts in esophagus, as identified by gene sequencing. Microcysts were present in 96.1% of the sheep, with a higher frequency (p < .05) in the heart (91.5%) compared to the esophagus (81.5%) samples. The microcysts were identified as Sarcocystis tenella by gene sequencing. Our results revealed a high frequency of Sarcocystis spp. infection in sheep from southern Brazil. To the authors knowledge, this is the first molecular confirmation of S. gigantea presence in Brazil.

Cross-sectional investigation on sheep sarcosporidiosis in Sardinia, Italy

An epidemiological survey on sarcosporidiosis was carried out with a cross sectional investigation on macroscopic and microscopic Sarcocystis spp. in Sarda breed sheep slaughtered in different abbatoirs of Sardinia, Italy. For the macroscopic survey, muscular samples (diaphragm, abdominal and intercostals muscles, cutaneous muscles and muscles of the thigh) from 769 slaughtered Sarda sheep, oesophagus (n=365) and laryngeal and pterygoid muscles (n=521) were macroscopically investigated and Polymerase Chain Reaction (PCR) on selected macroscopic cysts was performed for a molecular identification of macroscopic Sarcocystis species. For the microscopic investigation 112 heart samples from slaughtered Sarda sheep were collected and investigated with two different protocols: unstained (compression) examination and a molecular technique. The overall prevalence of infection for macroscopic forms of sarcocysts was of 23.3% (179/769) with prevalences higher in the oesophagus (31.6%; 125/395) compared with the other investigated tissue type; two different morphotypes, classified as large oval (LO) macroscopic cysts, identified as Sarcocystis gigantea, and slender fusiform (SF) sarcocysts, were identified. The examination of heart samples revealed an overall prevalence of 77.7% (87/112) for Sarcocystis spp.; the nested-PCR analysis of heart samples allowed to identify the microscopic species, Sarcocystis tenella and Sarcocystis arieticanis, with prevalences of 95.5% (107/112) and 17.8% (5/112) respectively. Reported results highlight the high prevalence of Sarcocystis infection in the island and suggests the need of an improvement of control and prevention strategies for this parasitosis.

Natural Sarcocystis gigantea infection in sheep from Southern Brazil

ABSTRACT: Protozoal diseases caused by species of Sarcocystiscan cause serious damage in sheep flocks, inducing decreased growth conversion rates and partial or complete loss of carcasses at the slaughterhouse. This article describes an outbreak of Sarcocystis giganteainfection in sheep slaughtered in a farm in Rio Grande do Sul State, Brazil. Between July and September 2013, three sheep showed multiple nodules in the esophagus that were microscopically characterized as encapsulated cysts filled with elongated, basophilic, nucleated structures morphologically consistent with S. giganteabradyzoites. Diagnosis was made based on the epidemiological, macroscopic, and microscopic findings. This is the first report of this infection in sheep in Rio Grande do Sul and should be recognized by veterinarians, especially during meat inspection.

Survey of prevalence and genotyping of Sarcocystis parasite in sheep of North Khorasan province

Background & Objective: Sarcocystosis is caused by the genus of Sarcocystis, an intracellular protozoan parasite in the phylum Apicomplexa. More than one hundred species of Sarcocystis can infect domestic and wild animals. This parasite is pathogenic for animals such as cattle and sheep; it also causes gastrointestinal symptoms in humans. The objective of this study was to determine the prevalence and genotyping of Sarcocystis strains in North Khorasan province. Materials&Methods: In this study, 320 samples of liver, diaphragm, heart and muscles were collected from 80 sheep (40 males and 40 females).The histological study was conducted for finding Sarcocyst. Furthermore, for observation of sarcocystis bradyzoite, 70 g of each tissue were digested using tissue digestion. PCR analysis was performed on all samples. Sequencing was done on 12 PCR product. Also, genotyping was carried out by Blast search and the homology analysis on 18 sRNA region. Data were analyzed by SPSS 16 software. Results . Sarcocyts were seen using histological method. Tissue digestion method showed that all samples(100%)were infected with Sarcocystis. Genotyping of 12 samples proved that sarcocystis belonged to Sarcocystis tenella (75%), Sarcocystis moulei (8/33%), and Sarcocystis gigantea (16/66%). Conclusion : The prevalence of Sarcocystis infection was high in the tissues of sheep in North Khorasan province. Moreover, Sarcocystis tenella, Sarcocystis moulei and Sarcocystis gigantea were reported for the first time in this province and Sarcocystis tenella was the most common species. Keywords: Sarcocystis, Genotyping, sheep, North khorasan province

The survival of Sarcocystis gigantea sporocysts following exposure to various chemical and physical agents

Using in vitro excystation as a measure of viability, it was found that at 4°C Sarcocystis gigantea sp sporocysts survived considerably better in tap water (85% excystation after 174 days) than in either 2.5% potassium dichromate (15% excystation after 174 days) or 2% sulphuric acid (0% excystation after 5 days). Although they were able to resist 48 h suspension at room temperature in most laboratory reagents and disinfectants tested, six (sulphuric acid, ammonia, methanol, ethanol, potassium hydroxide, sodium hydroxide, Medol) had substantial sporocysticidal properties. Further investigation with three of these showed that sporocyst excystation was reduced from 65% to less than 10% following contact with 2.5% sulphuric acid for 1 h or with 2% ammonia or 4% Medol for 4 h. Sporocysts were either killed or had their ability to excyst severely impaired by heating to 60°C and 55°C for 5 and 60 min, respectively, by exposure to ultraviolet radiation at a dose of 4000 ET, or by prolonged storage in water at 24°C. Sporocysts exposed to either constant or intermittent freezing at −18°C suffered a comparatively slow decline in excystation rate with time, as did those subjected to desiccation. The duration of survival of desiccated sporocysts was inversely related to relative humidity and after 245 days at 33% relative humidity and temperatures of 15°C or 24°C, 60% of such sporocysts excysted.

Separation of Antigens from Sarcocystis Species Using Chromatofocusing

Crude antigen preparations from bradyzoites of Sarcocystis species exhibit a high degree of cross-reactivity with antisera against heterologous Sarcocystis species, preventing the development of a species-specific immunological test for sarcocystiosis. In this study, we fractionated bradyzoite-derived protein extracts from Sarcocystis tenella, Sarcocystis arieticanis, Sarcocystis gigantea, and Sarcocystis muris by chromatofocusing and obtained distinct protein elution profiles for each species. We then examined the isolated protein fractions for antigenicity with homologous and heterologous reference sera in an enzyme-linked immunosorbent assay. Whereas some antigenic fractions of bradyzoite proteins had equally high reactivity with the homologous and heterologous sera, the reactivity of other fractions was 3-38 times higher with homologous serum than with heterologous sera. Mice immunized with less cross-reactive protein fractions of S. gigantea and S. muris bradyzoites produced a specific immune serum. Thus, it is possible to isolate species-specific antigens from crude mixtures of bradyzoite-derived Sarcocystis antigens for development of species-specific immunological tests for sarcocystiosis.

Activation of human CD4+ and CD8+ cells bySarcocystis gigantea lectin

As recently reported, Sarcocystis gigantea lectin (SGL) is a powerful mitogen and a polyclonal activator (Syn. S. ovifelis) of human peripheral B-cells. In the present study we investigated the reactivity of human T-helper (CD4) and T-suppressor (CD8) cells to SGL. Mononuclear cells (MNCs) from five newborns and six adults were examined cytofluorometrically for the expression of cell-surface differentiation and activation antigens using a set of seven monoclonal antibodies. In all, 96% of cord-blood and 81% of adult CD4 cells expressed receptors for interleukin-2 (Tac+) after 64 and 164 h microgram/ml). The percentages of neonatal and adult Tac+ CD8 cells amounted to 67% and 59%, respectively. Major histocompatibility complex (MHC) class II antigens identified using the monoclonal anti-human leukocyte antigen (HLA)-DR antibody L243 were expressed on 30% (CD4) and 44% (CD8) of adult T-cells. Neonatal HLA-DR+ T-lymphocytes were not detectable. In parallel, functional tests were performed to examine cell proliferation and MNC antibody production.

The in vitro excystation of Sarcocystis gigantea sporocysts

Studies on the in vitro excystation of Sarcocystis gigantea sporocysts revealed that pretreatment before exposure to trypsin and bile was an essential prerequisite. However, in contrast to Sarcocystis tenella and Sarcocystis capracanis, incubation in cysteine hydrochloride under CO 2 was largely unsuccessful for excysting Sarcocystis gigantea: of the pretreatments testes, only exposure to sodium hypochlorite proved effective. Excystation from sodium hypochlorite-pretreated S. gigantea sporocysts took place in trypsin and bile between temperatures of 30 and 43°C and occurred rapidly at 39°C. While the presence of bile or bile salts was essential for this process, that of trypsin was not, although more sporocysts excysted in its presence than in its absence. Excystation occurred in the presence of all bile types tested but not when Tween 80 was substituted for bile. The highest levels of excystation were recorded when cattle or sheep bile or sodium taurocholate were used and the lowest when chicken or pig bile were employed. Neither the concentration of sheep bile above 2.5%, nor hydrogen ion concentration (pH range 5.0–10.0) appeared to have any marked effect on the level of excystation obtained.

Production ofSarcocystis giganteasporocysts by experimentally infected cats

Abstract A study of 28 infections in 22 individually housed cats fed excised cysts of Sarcocystis gigantea from naturally infected sheep, revealed a consistent pattern of sporocyst shedding. In all cats, sporocyst excretion commenced 10 or 11 days postinfection (PI). Peak production occurred between 13 and 22 days PI, in most instances on Days 17and 18. Peaknumbers (rounded) rangedfrom 550 to 260 000 (mean = 53 000) sporocysts per gram of faeces or from 38 000 to 6.6 million (mean = 1.7 million) sporocysts per day. The number of days sporocysts were shed ranged from 26 to at least 60 days PI but in 26 of the 28 infections examined, > 80% of the total sporocyst yield was produced within 30 days of infection. The total numbers of sporocysts produced by individual cats over the patent period ranged from 164 000 to 56.6 million (mean = 12.7 million). These numbers tended to increase with increasing infective dose and tobe greater in those cats receiving multiple rather than equivalent single doses. Neither th...

Sarcocystis gigantea lectin ? mitogen and polyclonal B-cell activator

The present study further examined the in vitro response of human mononuclear cells (MNC) to the Sarcocystis gigantea lectin (SGL). The results confirm our previous report that SGL is mitogenic for human MNC. We now report that SGL is not only a potent mitogen but also a polyclonal activator for human peripheral B cells. As was true for pokeweed mitogen (PWM, 2 micrograms/ml), the addition of SGL (25 micrograms protein/ml) to cultures of MNC caused lymphocyte proliferation and B-cell maturation, indicated by a marked increase in IgG and IgM production. As measured by the [3H]-thymidine incorporation assay, SGL induced significantly higher proliferative responses than PWM (P less than 0.01, n = 24). The values obtained by SGL and PWM for IgG and IgM synthesis were essentially identical. As opposed to SGL, the sarcotoxin-containing fraction (SGTF) did not induce antibody formation or proliferative responses in human MNC.

Interactions betweenSarcocystis gigantea lectin and toxin-containing fractions in human lymphocyte cultures

Sarcocystis gigantea extract (SGE) was separated by affinity chromatography into one lectin-containing fraction (SGL) that was mitogenic to mononuclear cells (MNC) and another that lacked this lectin activity. The SGL-depleted Sarcocystis extract (SGTF) contained the so-called Sarcotoxin, inducing only a slight increase in MNC proliferation. Furthermore, preincubation of MNC with SGTF for 60 min suppressed the mitogenic capacity of SGL by 60%-90%. The results presented indicate that SGTF interacts with human MNC differently than SGL, particularly by interfering with the mitogenic lectin. These findings suggest that SGL and SGTF may be involved in different immunomechanisms induced by the parasite.

Evaluation of a concentration method for counting sarcocystis gigantea sporocysts in cat faeces

A technique for determining the numbers of S. gigantea sporocysts in cat faeces using a concentration procedure and haemocytometer was evaluated. The results showed that it was more accurate than a modified McMaster method and had a mean recovery rate of 73% at four levels of infection ranging from about 2000 to over 20 000 sporocysts per gram of faeeces.