RESUMEN
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.
Asunto(s)
Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Sarcocystis , Sarcocistosis , Sensibilidad y Especificidad , Enfermedades de las Ovejas , Animales , Sarcocystis/genética , Sarcocystis/aislamiento & purificación , Sarcocystis/clasificación , Ovinos , Sarcocistosis/veterinaria , Sarcocistosis/diagnóstico , Sarcocistosis/parasitología , Enfermedades de las Ovejas/parasitología , Enfermedades de las Ovejas/diagnóstico , Técnicas de Amplificación de Ácido Nucleico/métodos , Técnicas de Amplificación de Ácido Nucleico/veterinaria , Técnicas de Diagnóstico Molecular/métodos , ARN Ribosómico 28S/genética , ADN Protozoario/genéticaRESUMEN
Sarcocystis is a genus of protozoa with a worldwide distribution infecting a wide range of animals, including humans. Wild boars can harbor at least two species of Sarcocystis, that is, the zoonotic Sarcocystis suihominis, using humans as definitive hosts, and Sarcocystis miescheriana, for which wild and domestic canids serve as definitive hosts. In Portugal, hunting holds significant economic and social importance, and wild boars are among the most appreciated hunted species. As the consumption of wild boar meat can expose humans to several foodborne pathogens, the presence of trained hunters can make a difference in ensuring animal health surveillance and food safety. Herein, we report the detection of macroscopic cystic lesions associated with S. miescheriana in a wild boar hunted for human consumption, resulting in carcass condemnation. To the best of the authors' knowledge, the presence of S. miescheriana in wild boar tissues has never been associated with macroscopic pathological alterations before. Although S. miescheriana cannot infect humans, carcasses affected by grossly visible pathological changes must be declared unfit for consumption. Therefore, our finding points out the potential economic damage associated with carcass rejection due to the presence of gross lesions associated with generalized sarcocystosis. Nonetheless, further studies are required to investigate these alterations that currently appear to be occasional findings.
Asunto(s)
Sarcocystis , Sarcocistosis , Sus scrofa , Enfermedades de los Porcinos , Animales , Sarcocystis/aislamiento & purificación , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sus scrofa/parasitología , Enfermedades de los Porcinos/parasitología , Porcinos , Portugal , Inocuidad de los Alimentos , Humanos , Carne/parasitologíaRESUMEN
Muscles of 25 bobcats (Lynx rufus) from remote areas of Mississippi in 2017 were tested for parasites. Testing for Sarcocystis infections included microscopic examination of fresh unstained muscle squashes, pepsin digestion of hearts and tongues, and histological sections of paraffin-embedded tissues. Sarcocystis spp. infections were detected in the muscles of 21 (84%) by a combination of methods. Sarcocysts were detected in the unstained tongue squashes of 2 bobcats. Sarcocystis sp. bradyzoites were detected in the pepsin digests of 3 of 19 hearts, and 12 of 19 tongues. In paraffin-embedded histological sections, sarcocysts were detected in 7 of 25 hearts, 17 of 25 tongues, and 5 of 23 limb muscles. Based on the character of the cyst wall, at least 3 morphologic types of sarcocysts were detected: those with small spikes on the cyst wall, corresponding to Sarcocystis felis, those with long villar protrusions, corresponding to Sarcocystis neurona, and those lacking visible cyst wall protrusions, representing an unidentified type of sarcocyst. Myositis associated with sarcocysts was seen in the tongues of 3, and in the limb muscles of 1 bobcat. Multilocus genotyping of the DNA extracted from paraffin-embedded sections from 2 bobcats, employing 18S, 28S, COI, ITS-1, and 5.8S and rpoB genes, diagnosed Sarcocystis caninum, S. felis, Sarcocystis lutrae, and S. neurona. An encapsulated species of Trichinella was identified in the tongue of 1; it represents the first documented occurrences in bobcats from Mississippi. Taken together, these observations suggest intensive exposure of these wild carnivores to Trichinella tissue cysts, implies predation or scavenging on these tissues promotes parasite transmission, and raises caution concerning zoonotic risk when such meat is rendered for human consumption.
Asunto(s)
Lynx , Sarcocystis , Sarcocistosis , Lengua , Trichinella , Triquinelosis , Animales , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocystis/clasificación , Sarcocystis/aislamiento & purificación , Sarcocystis/genética , Lynx/parasitología , Mississippi , Triquinelosis/veterinaria , Triquinelosis/parasitología , Trichinella/aislamiento & purificación , Trichinella/clasificación , Trichinella/genética , Lengua/parasitología , Femenino , Masculino , Corazón/parasitología , Músculo Esquelético/parasitología , ADN Protozoario/aislamiento & purificación , ADN Protozoario/química , PrevalenciaRESUMEN
Sarcocystis bertrami (synonym: Sarcocystis fayeri) is a coccidian parasite that infects horses and donkeys in several countries. Dogs are known as definitive hosts of the parasite, however, the patent period is not well defined, and S. bertrami shed by dogs has never been confirmed by molecular methods. Here we investigated the shedding of S. bertrami by experimentally infected dogs and examined the excreted parasites by morphological and molecular tools. Three dogs of small breeds (one Yorkshire terrier and two miniature Pinschers) were acquired with ages of 30 and 60 days and were exclusively fed commercial dog food. Two dogs consumed equine muscle tissues containing cysts of S. bertrami. The third dog served as negative control and was simultaneously fed commercial dog food. The two animals that received equine tissues shed sporocysts and/or oocysts in their feces after prepatent periods of 13 and 23 days. The patent periods were 47 and 14 days. Sporocysts showed average dimensions of 14.19⯵m (± 0.53) x 10.06⯵m (± 0.44). The control dog did not shed sporocysts or oocysts of the parasite. Interestingly, patent periods had never been reported, and for one dog, the patent period (47 days) was longer than that reported for other Sarcocystidae parasites. PCRs to the gene 18S and to the internal transcribed spacer 1 (ITS1) of the rDNA were successfully performed with DNA extracted from sporocysts. ITS1 sequences were also obtained from the equine tissue cysts used to infect the dogs. Nucleotide sequences of cloned fragments of 18S from sporocysts, and ITS1 from both stages (tissue cysts and sporocysts) matched with S. bertrami (18S: 97.50-99.88â¯%; ITS1: 88.76-95.21â¯%), although high molecular diversity was observed with data from these loci. PCR to cox1 using sporocysts' DNA failed to amplify any product. The possibility of the existence of an additional and undescribed Sarcocystis species in the excreted sporocysts, besides S. bertrami, cannot be excluded from this experiment. To our knowledge, this is the first molecular confirmation of S. bertrami in canine feces. Sporocyst dimensions and prepatent periods observed in this study were similar to those previously described for S. bertrami and S. fayeri. In conclusion, the molecular, morphological and biological data generated here fit in previous descriptions for both S. bertrami and S. fayeri.
Asunto(s)
Enfermedades de los Perros , Heces , Oocistos , Sarcocystis , Sarcocistosis , Animales , Sarcocystis/genética , Sarcocystis/clasificación , Sarcocystis/aislamiento & purificación , Perros , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Enfermedades de los Perros/parasitología , Heces/parasitología , Caballos/parasitología , ARN Ribosómico 18S/genética , ADN Protozoario/genéticaRESUMEN
BACKGROUND: Sarcocystis is a food-borne zoonotic protozoan whose final hosts are humans, dogs, cats, and other carnivores and intermediate hosts are birds and mammals, especially humans and herbivores. Humans become infected by eating raw and undercooked meat contaminated with bradyzoites or by consuming water or food contaminated with the sporocyst stage of the parasite. OBJECTIVES: The aim of this study was to investigate the effects of gamma radiation and electron beam on the survival rate of Sarcocystis bradyzoites in infected beef and to determine the effective dose. METHODS: Three replicates of 100 g of infected meat were treated with different doses (0.5, 1, 1.5 and 2 kGy). As a control, 20 g of contaminated meat was stored separately at 4°C. The viability of the bradyzoites after digestion in pepsin solution was assessed, stained (trypan blue) and unstained, under a stereomicroscope. To assess survival of the bradyzoites, the irradiated meat samples were fed to 30 dogs. After 10 days, faecal samples were examined for sporocysts. RESULTS: The results showed that the highest and lowest mortality rate of Sarcocystis bradyzoites in infected organs using electron beam at a dose of 2 kGy were 92.5% and 100%, respectively, and the lowest mortality rate at a dose of 0.5 kGy were 2.5% and 7.89%, respectively. CONCLUSION: The results of statistical analysis showed that the mortality rate of Sarcocystis bradyzoites was significant between different doses of gamma ray and electron beam, so that gamma rays were better compared to electron beam in destroying Sarcocystis bradyzoites.
Asunto(s)
Sarcocystis , Sarcocystis/efectos de la radiación , Sarcocystis/fisiología , Animales , Bovinos , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Carne Roja/parasitología , Rayos gamma , Perros , Irradiación de Alimentos , Relación Dosis-Respuesta en la Radiación , Enfermedades de los Bovinos/parasitología , ElectronesRESUMEN
Sarcocystis miescheriana infection is an important cause of carcass condemnation during meat inspection. The infection can cause morbidity and mortality in domestic pigs. In this study, an 8-month-old finisher pig was presented to a local abattoir for slaughter. Multiple white nodular lesions affecting the meat were observed, resulting in the condemnation of the carcass. Consequently, half of the carcass was submitted to the necropsy diagnostic laboratory in the School of Veterinary Medicine for further evaluation. Grossly, all superficial and deep muscle groups had severe multifocal macrocysts (3 mm × 2 mm × 1 mm) on the surface and extending deep into the skeletal musculature. Histopathology revealed moderate multifocal granulomatous and eosinophilic myositis with intralesional degenerated and intact parasites. Sample genomic DNA sequence analysis of the 18S RNA gene showed 100% identity to S. miescheriana in the GenBank. This is the first report of S. miescheriana in Grenada, West Indies.
Asunto(s)
Sarcocystis , Sarcocistosis , Enfermedades de los Porcinos , Animales , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocystis/aislamiento & purificación , Sarcocystis/genética , Enfermedades de los Porcinos/parasitología , Enfermedades de los Porcinos/patología , Porcinos , Grenada/epidemiología , Sus scrofaRESUMEN
Several wild game meat species, including deer and feral pigs are hunted and consumed in Australia. Feral pigs and deer are not indigenous to Australia, but they have proliferated extensively and established their presence in every state and territory. Following the report of a sambar deer displaying Sarcocystis like white cysts in its rump muscles, the present study was conducted to explore the prevalence of Sarcocystis infections in wild deer and feral pigs in the southeastern regions of Australia. Oesophagus, diaphragm, and heart tissue from 90 deer and eight feral pigs were examined visually for sarcocysts. All results were negative. PCR testing of randomly selected deer and feral pigs yielded positive results, which were subsequently supported by histopathology. This is the first study to report the presence of Sarcocystis spp. in deer and feral pigs in Australia. As no visual cysts were found on the heart or oesophagus that came back positive with PCR, infected animals, particularly those reared free-range, could be passing through meat quality checks unidentified. If people consume this meat without cooking it properly, it may lead to a human infection of Sarcocystis. However, a more targeted study focused on determining the parasite's prevalence and assessing its risks is necessary to determine if it constitutes a food safety issue. As this species has been found not only in feral pigs but also in domestic pigs, the potential for infection spreading between feral pigs and pigs in free-range livestock systems is high, potentially posing a large problem for the Australian pork industry, particularly with the increased emphasis on free-range pig husbandry. Future studies should concentrate on determining the species of Sarcocystis in feral animals commonly consumed as game meat to determine potential zoonotic risks. This could also include a more in-depth look at the prevalence of Sarcocystis infections in other game animals. Identifying where these parasites are present and to what extent, are important areas for future studies.
Asunto(s)
Animales Salvajes , Ciervos , Carne , Sarcocystis , Sarcocistosis , Enfermedades de los Porcinos , Animales , Sarcocystis/aislamiento & purificación , Sarcocystis/genética , Sarcocystis/clasificación , Ciervos/parasitología , Australia/epidemiología , Porcinos , Sarcocistosis/epidemiología , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Animales Salvajes/parasitología , Enfermedades de los Porcinos/parasitología , Enfermedades de los Porcinos/epidemiología , Carne/parasitología , Prevalencia , HumanosRESUMEN
Equine protozoal myeloencephalitis (EPM) is a challenging disease to diagnose in horses with neurological signs. To optimize contemporary diagnostic testing, including the use of serum:CSF antibody ratios, the SarcoFluor antibody test for Sarcocystis neurona requires revalidation. The SarcoFluor, a previously validated immunofluorescent antibody test (IFAT) for the detection of antibodies specific to S. neurona in serum and cerebrospinal fluid (CSF) of naturally infected horses was analyzed using recent data and considering a serum:CSF antibody ratio threshold. Utilization of serum and CSF phosphorylated neurofilament heavy protein (pNfH) concentrations in support of an EPM diagnosis was also evaluated. 172 horses were divided into three groups: EPM-positive horses (EPM+, n=42), neurological non-EPM horses (n=74) confirmed with non-EPM neurological diseases (cervical vertebral compressive myelopathy, equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy), and control horses (control, n=56) without neurological signs and neurological abnormalities on histology. Logistic regression was used to compare EPM diagnostic regimens. Specifically, EPM+ horses were compared with neurological non-EPM horses showing neurological signs. To consider diagnostic utility, post-test probabilities were calculated by titer. When differentiating between EPM and other neurological diseases, the combination of serum and CSF SarcoFluor testing added more information to the model accuracy than either test alone. Using serum and CSF for pNfH in support of an EPM diagnosis did not identify cutoffs with statistically significant odds ratios but increased the overall model accuracy when used with the IFAT. Utilization of IFAT titers against S. neurona in serum and CSF result in a high post-test probability of detecting EPM+ horses in a clinical setting.
Asunto(s)
Anticuerpos Antiprotozoarios , Enfermedades de los Caballos , Sarcocystis , Sarcocistosis , Animales , Caballos , Sarcocystis/inmunología , Anticuerpos Antiprotozoarios/sangre , Anticuerpos Antiprotozoarios/líquido cefalorraquídeo , Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/parasitología , Enfermedades de los Caballos/líquido cefalorraquídeo , Sarcocistosis/veterinaria , Sarcocistosis/diagnóstico , Sarcocistosis/parasitología , Sensibilidad y Especificidad , Técnica del Anticuerpo Fluorescente/veterinaria , Encefalomielitis Equina/veterinaria , Encefalomielitis Equina/diagnóstico , Encefalomielitis Equina/parasitología , Encefalomielitis/veterinaria , Encefalomielitis/parasitología , Encefalomielitis/diagnóstico , Encefalomielitis/líquido cefalorraquídeoRESUMEN
Sarcocystis spp. are cyst-forming coccidia characterized by a two-host predator-prey life cycle. Sarcocysts are formed in muscles or nervous system of the intermediate host, while sporocysts develop in the small intestine of the definitive host. The intermediate hosts of Sarcocystis falcatula are wild birds. Colombia is one of the countries with the greatest biodiversity of birds, however, there are few studies related to this parasite in wild birds. This study presents the morphological and molecular detection of Sarcocystis falcatula collected from the emerald toucanet (Aulacorhynchus albivitta), a wild bird species endemic to South America. Pectoral muscle samples were obtained, and microscopic and molecular detection was performed by light microscopy, transmission electron microscopy, and amplifying of the first internal transcribed spacer (ITS-1) and surface antigen-encoding genes (SAGs). Sarcocystis measured an average of 161 × 42 µm, with a cyst wall â¼0.4 µm thick. Ultrastructurally, the sarcocyst wall type 11b-like consisted of numerous villar protrusions of 850 nm wide on average. The ITS-1 sequence showed 97.0-99.7% identity to S. falcatula previously described from birds in the United States and Brazil, respectively. Concatenated phylogenetic analysis based on SAG2, SAG3 and SAG4 confirmed that the new isolate is grouped with other sequences of Sarcocystis from South America, but divergent from those isolates obtained in North America. The results of this study demonstrate for the first time the presence of S. falcatula in a wild bird from Colombia.
Asunto(s)
Enfermedades de las Aves , Sarcocystis , Sarcocistosis , Animales , Sarcocystis/genética , Sarcocystis/clasificación , Sarcocystis/aislamiento & purificación , Sarcocystis/ultraestructura , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocistosis/epidemiología , Colombia , Enfermedades de las Aves/parasitología , Filogenia , Microscopía Electrónica de Transmisión/veterinaria , ADN Protozoario/análisis , Falconiformes/parasitologíaRESUMEN
Sarcocystis spp. cause pigeon protozoan encephalitis, a neuronal disease. A female pigeon exhibiting torticollis had a necrotic area in the cerebral hemisphere surrounded by lesions with perivascular cuffing, gliosis, granulomatous foci, and meningitis. Non-necrotic lesions were also observed in the brainstem. Intact and degenerative schizonts were observed within the neuropils and neurons in the lesions. Deoxyribonucleic acid (DNA) was extracted from paraffin-embedded brain tissues and genetically analyzed after gel electrophoresis to determine Sarcocystis spp. using specific primer sets for 28S ribosomal ribonucleic acid and internal transcribed spacer region-1. DNA sequencing confirmed a significant homology with S. calchasi. This is the first report of meningoencephalitis with malacia caused by S. calchasi in a rock pigeon in Japan.
Asunto(s)
Enfermedades de las Aves , Columbidae , Meningoencefalitis , Sarcocystis , Sarcocistosis , Animales , Sarcocystis/aislamiento & purificación , Sarcocystis/genética , Columbidae/parasitología , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocistosis/patología , Femenino , Japón , Enfermedades de las Aves/parasitología , Enfermedades de las Aves/patología , Meningoencefalitis/veterinaria , Meningoencefalitis/parasitología , Meningoencefalitis/patología , Encéfalo/patología , Encéfalo/parasitologíaRESUMEN
The genus Sarcocystis includes protozoan parasites with an indirect life cycle. Sarcocystis spp. can infect various animal species and humans, causing sarcocystosis, a parasitosis of economic importance and zoonotic concern. Wild boars can act as intermediate hosts for Sarcocystis miescheriana and the zoonotic Sarcocystis suihominis that infects humans by consumption of raw or undercooked infected swine meat. In the present study, the diaphragmatic muscle tissue of 123 wild boars hunted in Greece was examined to determine the frequency of Sarcocystis spp. The samples were examined by tissue compression and molecular techniques. Under light microscopy, 34 out of 123 (27.6%) wild boars tested positive for Sarcocystis spp., while a higher infection prevalence (75%) was revealed by multiplex PCR performed in 100 of the samples. The partial mtDNA cox1 gene (~ 1100 bp) of 20 samples tested positive for S. miescheriana by multiplex PCR was amplified and sequenced. Sarcocystis miescheriana was identified as the only species involved in these infections. This is the first study on the prevalence of Sarcocystis spp. in wild animals in Greece. Further, large-scale surveys are needed to assess the prevalence and species of this parasite in Greece and to design efficient control and preventive measures in a One Health perspective.
Asunto(s)
Sarcocystis , Sarcocistosis , Sus scrofa , Enfermedades de los Porcinos , Animales , Sarcocystis/genética , Sarcocystis/aislamiento & purificación , Sarcocystis/clasificación , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocistosis/epidemiología , Grecia/epidemiología , Sus scrofa/parasitología , Enfermedades de los Porcinos/parasitología , Enfermedades de los Porcinos/epidemiología , Porcinos , ADN Protozoario/genética , Microscopía , Prevalencia , Análisis de Secuencia de ADN , ADN Mitocondrial/genética , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Complejo IV de Transporte de Electrones/genética , Diafragma/parasitologíaRESUMEN
Asexual replication in the apicomplexan Sarcocystis neurona involves two main developmental stages: the motile extracellular merozoite and the sessile intracellular schizont. Merozoites invade host cells and transform into schizonts that undergo replication via endopolygeny to form multiple (64) daughter merozoites that are invasive to new host cells. Given that the capabilities of the merozoite vary significantly from the schizont, the patterns of transcript levels throughout the asexual lifecycle were determined and compared in this study. RNA-Seq data were generated from extracellular merozoites and four intracellular schizont development time points. Of the 6,938 genes annotated in the S. neurona genome, 6,784 were identified in the transcriptome. Of these, 4,111 genes exhibited significant differential expression between the merozoite and at least one schizont development time point. Transcript levels were significantly higher for 2,338 genes in the merozoite and 1,773 genes in the schizont stages. Included in this list were genes encoding the secretory pathogenesis determinants (SPDs), which encompass the surface antigen and SAG-related sequence (SAG/SRS) and the secretory organelle proteins of the invasive zoite stage (micronemes, rhoptries, and dense granules). As anticipated, many of the S. neurona SPD gene transcripts were abundant in merozoites. However, several SPD transcripts were elevated in intracellular schizonts, suggesting roles unrelated to host cell invasion and the initial establishment of the intracellular niche. The hypothetical genes that are potentially unique to the genus Sarcocystis are of particular interest. Their conserved expression patterns are instructive for future investigations into the possible functions of these putative Sarcocystis-unique genes. IMPORTANCE: The genus Sarcocystis is an expansive clade within the Apicomplexa, with the species S. neurona being an important cause of neurological disease in horses. Research to decipher the biology of S. neurona and its host-pathogen interactions can be enhanced by gene expression data. This study has identified conserved apicomplexan orthologs in S. neurona, putative Sarcocystis-unique genes, and gene transcripts abundant in the merozoite and schizont stages. Importantly, we have identified distinct clusters of genes with transcript levels peaking during different intracellular schizont development time points, reflecting active gene expression changes across endopolygeny. Each cluster also has subsets of transcripts with unknown functions, and investigation of these seemingly Sarcocystis-unique transcripts will provide insights into the interesting biology of this parasite genus.
Asunto(s)
Merozoítos , Sarcocystis , Sarcocystis/genética , Sarcocystis/crecimiento & desarrollo , Merozoítos/crecimiento & desarrollo , Esquizontes/genética , Esquizontes/crecimiento & desarrollo , Proteínas Protozoarias/genética , Proteínas Protozoarias/metabolismo , Transcriptoma , Perfilación de la Expresión Génica , Reproducción Asexuada/genética , Animales , Sarcocistosis/parasitología , Sarcocistosis/veterinaria , Estadios del Ciclo de Vida/genéticaRESUMEN
A senile male black capuchin monkey (Sapajus nigritus) kept under human care in a Zoo was found dead after 2 weeks presenting signals of weight loss and hyporexia. Histopathological revealed a necrotizing encephalitis. Although it was not observed microscopically, Sarcocystis sp infection was detected in brain tissue from molecular assays. These infections have been rarely described in neotropical primates, particularly associated with tissue lesions.
Asunto(s)
Enfermedades de los Monos , Sarcocystis , Sarcocistosis , Animales , Sarcocistosis/veterinaria , Sarcocistosis/diagnóstico , Sarcocistosis/parasitología , Sarcocystis/aislamiento & purificación , Sarcocystis/genética , Enfermedades de los Monos/parasitología , Enfermedades de los Monos/diagnóstico , Masculino , Animales de Zoológico , Resultado Fatal , Encefalitis/veterinaria , Encefalitis/parasitología , Encefalitis/diagnóstico , SapajusRESUMEN
Currently, research on apicomplexan Sarcocystis parasites is mainly carried out by analyzing animal carcasses. However, environmental studies would not only allow faster detection of possible sources of infection but also avoid the use of animals for investigations. Therefore, in the current study, we aimed to identify tested Sarcocystis species in sediment collected from water bodies located in the southeastern Baltic countries. A total of 99 sediment samples were collected during the summer from different types of water bodies in Estonia, Latvia, Lithuania, and Poland. Species-specific nested PCR targeting cox1 gene was used for the detection of selected Sarcocystis species (S. cruzi, S. bovifelis, S. hirsuta, S. arieticanis, S. tenella, S. capracanis, S. miescheriana, and S. bertrami) infecting livestock. The results showed a statistically lower (p < 0.05) occurrence of Sarcocystis parasites in Estonia (50%) compared to three countries, where the detection rate of Sarcocystis spp. DNA was remarkably higher, ranging from 88 to 100%. Among Sarcocystis species tested, S. cruzi (83.8%) and S. arieticanis (55.6%) using cattle and sheep as their intermediate hosts were most commonly identified. The detection rates of some of the analyzed Sarcocystis species were significantly different in southeastern Baltic countries. It is discussed that the detection rates of certain Sarcocystis species depend not only on the number of animals per 1 km2 but also on various ecological factors and farming practices that differ in the amount of contact domestic animals have with predators and the potential for animals to become infected through natural water or food sources.
Asunto(s)
Ecosistema , Sedimentos Geológicos , Sarcocystis , Sarcocystis/genética , Sarcocystis/aislamiento & purificación , Sarcocystis/clasificación , Animales , Sedimentos Geológicos/parasitología , Polonia , Ovinos , Reacción en Cadena de la Polimerasa , Sarcocistosis/parasitología , Sarcocistosis/veterinaria , Sarcocistosis/epidemiología , Bovinos , Lituania/epidemiología , Países Bálticos , Biodiversidad , ADN Protozoario/genética , Letonia/epidemiología , EstoniaRESUMEN
Sarcocystis spp. are protozoan parasites that form cysts in the organs and musculature of various animal species. The species Sarcocystis miescheriana and Sarcocystis suihominis are pathogenic to pigs and wild boars (Sus scrofa), acting as intermediate hosts, while humans are the definitive host for S. suihominis. To date, there have been no reports of the identification of these coccidian species in Sus scrofa in Brazil. Therefore, in this study, we conducted the first molecular identification of Sarcocystis species using PCR-RFLP and sequencing. A total of 210 samples were analyzed, of this total, 67 tested positive for Sarcocystis spp., representing 31.9% of the total samples assessed. Out of the total positive samples, 55 (82.1%) were identified as S. miescheriana and 8 (11.9%) as S. suihominis, a zoonotic species. Additionally, other species related to bovines, such as S. cruzi and zoonotic S. hominis, were detected in 3.0% of the samples, serving as contaminants in the pork products. The presence of S. suihominis in swine and wild boar samples is concerning due to the zoonotic risk and potential environmental contamination, as humans act as definitive hosts, also for the presence of S. hominis as a bovine contaminant in pork sausages. Furthermore, we confirmed the efficacy of the PCR-RFLP technique as a reliable tool for the identification of Sarcocystis species, demonstrating its potential use in laboratories for molecular diagnosis and rapid identification of these parasites, aiming to protect public health and ensure food safety.
Asunto(s)
Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Sarcocystis , Sarcocistosis , Sus scrofa , Enfermedades de los Porcinos , Animales , Sarcocystis/genética , Sarcocystis/aislamiento & purificación , Sarcocystis/clasificación , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocistosis/epidemiología , Brasil/epidemiología , Sus scrofa/parasitología , Enfermedades de los Porcinos/parasitología , Enfermedades de los Porcinos/epidemiología , Porcinos , Reacción en Cadena de la Polimerasa/veterinariaRESUMEN
Sarcocystis spp. are apicomplexan cyst-forming parasites that can infect numerous vertebrates, including birds. Sarcosporidiosis infection was investigated in three muscles (breast, right and left thigh muscle) and one organ (heart) of four Razorbill auks (Alca torda) stranded between November and December 2022 on the shores of the Mediterranean Sea in Nabeul and Bizerte governorates, Northern Tunisia. Two of the four tested A. torda were PCR positive for 18S rRNA Sarcocystis spp. gene. Among the examined 16 muscles/organs, only one breast and one right thigh were Sarcocystis spp. PCR-positive (12.5% ± 8.3, 2/16). Our results showed a relatively high molecular prevalence of Sarcocystis spp. in Razorbill auks (A. torda). Sarcocystis spp. sequence described in the present study (GenBank number: OR516818) showed 99.56-100% identity to Sarcocystis falcatula. In conclusion, our results confirmed the infection of Razorbill auks (A. torda) by S. falcatula. Further research is needed on different migratory seabirds' species in order to identify other Sarcocystis species.
Asunto(s)
ARN Ribosómico 18S , Sarcocystis , Sarcocistosis , Sarcocystis/genética , Sarcocystis/aislamiento & purificación , Sarcocystis/clasificación , Animales , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocistosis/epidemiología , Túnez/epidemiología , Mar Mediterráneo , ARN Ribosómico 18S/genética , Enfermedades de las Aves/parasitología , Enfermedades de las Aves/epidemiología , ADN Protozoario/genética , Filogenia , Charadriiformes/parasitología , Reacción en Cadena de la Polimerasa , Prevalencia , Análisis de Secuencia de ADN , ADN Ribosómico/genética , ADN Ribosómico/químicaRESUMEN
In the present study, tissue samples (tongue, esophagus and heart) were investigated from dromedary camels of India for identification and characterization of Sarcocystis spp. using histopathology, PCR and gene sequencing. Genomic DNA extracted from these tissue samples was used for PCR amplification of the cytochrome c oxidase subunit I gene (cox1) of Sarcocystis spp. and the partial sequence of small subunit ribosomal RNA (18S rRNA) gene of the S. cameli. The PCR products were purified, sequenced and analyzed using bioinformatics tools. Based on phylogenetic analysis of the cox1 gene, the sequences of the present study clustered with those of S. cameli, hosted by dromedary camels of Iraq and a close association was observed with S. masoni hosted by dogs and alpacas of China. Until now, there are no 18S rRNA sequences of S. cameli available in GenBank and this is the first study recording 18S rRNA sequences of S. cameli which were grouped with S. masoni from alpaca of China and guanaco and llama of Argentina in phylogenetic analysis. These findings could be useful for further studies on the characterization through molecular epidemiology, genetic diversity and host specificity of S. cameli.
Asunto(s)
Camelus , Filogenia , ARN Ribosómico 18S , Sarcocystis , Sarcocistosis , Animales , Sarcocystis/genética , Sarcocystis/clasificación , Sarcocystis/aislamiento & purificación , Camelus/parasitología , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocistosis/epidemiología , ARN Ribosómico 18S/genética , India/epidemiología , Complejo IV de Transporte de Electrones/genética , Complejo IV de Transporte de Electrones/análisisRESUMEN
BACKGROUND: The geographic distribution and host-parasite interaction networks of Sarcocystis spp. in small mammals in eastern Asia remain incompletely known. METHODS: Experimental infections, morphological and molecular characterizations were used for discrimination of a new Sarcocystis species isolated from colubrid snakes and small mammals collected in Thailand, Borneo and China. RESULTS: We identified a new species, Sarcocystis muricoelognathis sp. nov., that features a relatively wide geographic distribution and infects both commensal and forest-inhabiting intermediate hosts. Sarcocystis sporocysts collected from rat snakes (Coelognathus radiatus, C. flavolineatus) in Thailand induced development of sarcocysts in experimental SD rats showing a type 10a cyst wall ultrastructure that was identical with those found in Rattus norvegicus from China and the forest rat Maxomys whiteheadi in Borneo. Its cystozoites had equal sizes in all intermediate hosts and locations, while sporocysts and cystozoites were distinct from other Sarcocystis species. Partial 28S rRNA sequences of S. muricoelognathis from M. whiteheadi were largely identical to those from R. norvegicus in China but distinct from newly sequenced Sarcocystis zuoi. The phylogeny of the nuclear 18S rRNA gene placed S. muricoelognathis within the so-called S. zuoi complex, including Sarcocystis attenuati, S. kani, S. scandentiborneensis and S. zuoi, while the latter clustered with the new species. However, the phylogeny of the ITS1-region confirmed the distinction between S. muricoelognathis and S. zuoi. Moreover, all three gene trees suggested that an isolate previously addressed as S. zuoi from Thailand (KU341120) is conspecific with S. muricoelognathis. Partial mitochondrial cox1 sequences of S. muricoelognathis were almost identical with those from other members of the group suggesting a shared, recent ancestry. Additionally, we isolated two partial 28S rRNA Sarcocystis sequences from Low's squirrel Sundasciurus lowii that clustered with those of S. scandentiborneensis from treeshews. CONCLUSIONS: Our results provide strong evidence of broad geographic distributions of rodent-associated Sarcocystis and host shifts between commensal and forest small mammal species, even if the known host associations remain likely only snapshots of the true associations.
Asunto(s)
Enfermedades de los Roedores , Sarcocystis , Sarcocistosis , Ratas , Animales , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , ARN Ribosómico 28S/genética , Reacción en Cadena de la Polimerasa , Ratas Sprague-Dawley , ARN Ribosómico 18S/genética , Filogenia , Sciuridae , Murinae , Enfermedades de los Roedores/parasitologíaRESUMEN
PURPOSE: Using molecular techniques, we have previously shown that carnivorous mammals of the family Mustelidae might be common definitive hosts for various protozoan Sarcocystis species. In the present study we aimed to unravel whether Sarcocystis species using ungulates as intermediate hosts and canids or felids as definitive hosts can be found in intestine of mustelids. METHODS: Small intestine samples of 93 individual mustelids of five different species from Lithuania were examined. Sarcocystis species were identified based on species-specific PCR and subsequent cox1 sequencing. RESULTS: Six Sarcocystis species (S. arieticanis, S. bertrami, S. capracanis, S. capreolicanis, S. linearis and S. morae) defined by ungulate-canid life cycle were detected for the first time in small intestines of mustelids. By contrast, the prevalence of Sarcocystis characterised by ungulate-felid life cycle was low (3.2%). Overall, 76% of the examined animals were positive for at least one of the studied Sarcocystis species. Four species, S. arieticanis, S. bertrami, S. capracanis and S. morae were most commonly found, with the detection rate of about 40%. CONCLUSIONS: The current finding, in addition to our previous studies, suggests that mustelids play an important role in the spread of various Sarcocystis species.
Asunto(s)
Intestino Delgado , Mustelidae , Sarcocystis , Sarcocistosis , Animales , Sarcocistosis/veterinaria , Sarcocistosis/parasitología , Sarcocystis/genética , Sarcocystis/clasificación , Sarcocystis/aislamiento & purificación , Intestino Delgado/parasitología , Mustelidae/parasitología , Lituania , Estadios del Ciclo de Vida , Reacción en Cadena de la Polimerasa , FilogeniaRESUMEN
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.