Dataset Information

Molecular Identification of Sarcocystis rileyi and Sarcocystis sp. (Closely Related to Sarcocystis wenzeli) in Intestines of Mustelids from Lithuania.

ABSTRACT: The genus Sarcocystis is a group of numerous protozoan parasites having a two-host life cycle. Based on laboratory experiments and/or phylogenetic analysis results it was shown that seven Sarcocystis spp. producing sarcocsyts in bird tissues are transmitted via predatory placental mammals. To date the role of small mammals of the family Mustelidae in the distribution of avian Sarcocystis spp. have not been studied. During the current investigation, intestinal mucosa scrapings of 115 mustelids belonging to five species were tested for S. albifronsi, S. anasi, S. rileyi, and S. wenzeli infecting anseriforms and chickens. Microscopically, free sporocysts, sporulating oocysts, and loose oocysts were found in 61 samples (53.0%). Using nested PCR targeting the ITS1 region and sequencing, S. rileyi was confirmed in eight American minks, two European polecats and single European badger. Sarcocystis sp. was identified in one American mink and one European pine marten. Based on the partial ITS1 region this parasite showed that 100% identity to pathogenic Sarcocystis sp. caused a fatal infection in backyard chickens from Brazil. Phylogenetically, the Sarcocystis sp. identified in our study was most closely related to S. wenzeli parasitising domestic fowl (Gallus domesticus).

SUBMITTER: Prakas P

PROVIDER: S-EPMC9913585 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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Molecular Identification of Sarcocystis rileyi and Sarcocystis sp. (Closely Related to Sarcocystis wenzeli) in Intestines of Mustelids from Lithuania.

Prakas Petras P Moskaliova Darija D Šneideris Donatas D Juozaitytė-Ngugu Evelina E Maziliauskaitė Evelina E Švažas Saulius S Butkauskas Dalius D

Animals : an open access journal from MDPI 20230129 3

The genus Sarcocystis is a group of numerous protozoan parasites having a two-host life cycle. Based on laboratory experiments and/or phylogenetic analysis results it was shown that seven Sarcocystis spp. producing sarcocsyts in bird tissues are transmitted via predatory placental mammals. To date the role of small mammals of the family Mustelidae in the distribution of avian Sarcocystis spp. have not been studied. During the current investigation, intestinal mucosa scraping ...[more]

PMID: 36766356

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Project description:We investigated the morphology and phylogenetic relationships of novel and previously recognized Sarcocystis spp. infecting small mammals and colubrid snakes in Asia. The nuclear 18S rRNA and mitochondrial cox1 of Sarcocystis sp.1 from mangrove snakes (Boiga dendrophila) in Thailand and Sarcocystis sp.2 from a ricefield rat (Rattus argentiventer) in Sumatra were partially sequenced. Sporocysts of Sarcocystis sp.1 induced development of sarcocysts in experimentally infected rats, which showed a unique ultrastructure that was observed previously by S.P. Kan in rats from Malaysia; therefore, we describe this species as Sarcocystis kani sp. nov. Its integration into the 18S rRNA phylogeny of Sarcocystis spp. cycling between small mammals and colubrid snakes helped clarify relationships among the so-called S. zuoi-complex of molecularly cryptic species: Sarcocystis kani sp. nov., S. sp.2, S. attenuati, S. scandentiborneensis, and S. zuoi were all included in this clade. Tree topology was resolved into dichotomies congruent with the morphological disparities between the taxa. However, cox1 gene sequencing (including newly sequenced S. singaporensis and S. zamani) revealed that Sarcocystis kani, S. attenuati, and S. scandentiborneensis were identical suggesting a recent, common ancestry. To identify other distinctive features, lineage-specific molecular patterns within both genes were examined revealing that all 18S rRNA sequences of the S. zuoi - complex possess a unique, 7-nt long motif in helix 38 of domain V7 that was different in S. clethrionomyelaphis which branched off basally from the complex. Three-dimensional homology modelling of COX1 protein structure identified amino acid substitutions within the barcode area specific for the S. zuoi-complex and substantial divergence in structurally important amino acids between Sarcocystis species of snakes as definitive hosts and other lineages of the Sarcocystidae. We discuss the utility of selected genes for species delimitation of the Sarcocystis spp. under investigation, which probably evolved during recent radiations of their intermediate and definitive hosts.

Project description:BackgroundSix Sarcocystis species are known to use cattle (Bos taurus) as the intermediate host, two of which, S. hominis and S. heydorni, are zoonotic. There is a need for a method that will enable rapid identification of the Sarcocystis species in cattle.MethodsThe diaphragm muscles of 102 cattle from Lithuania were examined for the presence of Sarcocystis spp., using two different methods for species identification. Individual sarcocysts were isolated from squash preparations of the diaphragm muscle under the light microscope, followed by genetic characterisation of excised cysts using sequence analysis of the 18S rRNA (18S rRNA) and cytochrome c oxidase subunit I (cox1) genes. The same cattle muscle samples were digested and species-specific PCR analyses targeting cox1 were developed to identify the Sarcocystis isolates to the species level.ResultsUnder the light microscope, sarcocysts were detected in 87.3% of animals, and Sarcocystis infection was verified in all digested samples. Three species, namely S. cruzi (n = 20), S. bovifelis (n = 23) and S. hirsuta (n = 6), were identified by DNA sequence analysis of isolated sarcocysts. Based on sequence analysis of cox1, the level of genetic variability depended on Sarcocystis species and geographical location. Four Sarcocystis species, S. cruzi (96.1%), S. bovifelis (71.6%), S. hirsuta (30.4%) and S. hominis (13.7%), were confirmed in the digested samples. In individual samples, the most common finding was two species of Sarcocystis (44.1%), followed by three species (26.5%), a single species (24.5%) and four species (4.9%).ConclusionsAlthough examination of tissue preparations under the light microscrope did not detect any sarcocysts belonging to S. hominis, this species was identified in the digested samples subjected to a cox1-specific PCR analysis. These results demonstrate the need for effective molecular diagnosis techniques to detect Sarcocystis spp., which may be present at a lower prevalence and not detectable among the limited number of sarcocysts identified individually under the light microscope.

Project description:BackgroundThe cyst-forming coccidia of the genus Sarcocystis (Sarcocystidae) are widespread protists of mammals, particularly of domestic and wild ruminants. Research on genus Sarcocystis in wild members of the subfamily Caprinae is, however, rather limited. Sarcocystis in the Alpine ibex (Capra ibex) have only been investigated in depth once and then solely by morphological techniques. In the current investigation we aimed to morphologically and genetically characterise Sarcocystis species of Alpine ibex in Austria.MethodsSarcocysts detected in the diaphragm and myocardium muscles were morphologically described using light microscopy and transmission electron microscopy (TEM). Isolated sarcocysts were molecularly identified and characterised at the level of the 18S ribosomal RNA (rRNA) gene and cytochrome c oxidase I gene (cox1). The obtained sequences were subjected to phylogenetic analysis.ResultsThree Sarcocystis species, namely S. capracanis, S. cornagliai and S. rossii n. sp., were found in Alpine ibex. For the first time, we genetically characterised S. cornagliai, which is most closely related to Sarcocystis species that are transmitted by corvid birds. Sarcocysts of S. rossii n. sp. were found to be ribbon-shaped, with pointed tips. Hair-like protrusions about 5 μm in length were observed on sarcocyst walls. Observation of toluidine blue-stained semi-thin sections revealed that the sarcocyst of S. rossii n. sp. was thin-walled. Using TEM, cyst walls were observed to be similar to type 7a, with thin hair-like villar protrusions on the cyst wall, which were filled with many fine electron-dense granules. The ground substance layer was particularly thin, measuring 0.2-0.4 μm. The cox1 sequences of S. rossii n. sp. had the highest similarity to those of Sarcocystis arieticanis and Sarcocystis hircicanis. Sarcocystis rossii n. sp. had a close phylogenetic relationship with species that use canids as definitive hosts.ConclusionsThis study confirms the role of the Alpine ibex as an intermediate host of three Sarcocystis species and sets a new host record for S. capracanis. It also provides the first molecular data on Sarcocystis from Alpine ibex and on S. cornagliai. In addition, a new species, S. rossii, was identified and described. Phylogenetic analyses suggested corvid birds and canids as potential definitive hosts for S. cornagliai and S. rossii n. sp., respectively.

Dataset Information

Molecular Identification of Sarcocystis rileyi and Sarcocystis sp. (Closely Related to Sarcocystis wenzeli) in Intestines of Mustelids from Lithuania.

Publications

Molecular Identification of <i>Sarcocystis rileyi</i> and <i>Sarcocystis</i> sp. (Closely Related to <i>Sarcocystis wenzeli</i>) in Intestines of Mustelids from Lithuania.

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