Sarcocystis entzerothi n. sp. from the European roe deer (Capreolus capreolus)

Prakas, Petras; Rudaitytė, Eglė; Butkauskas, Dalius; Kutkienė, Liuda

doi:10.1007/s00436-016-5288-7

Cited by 22 publications

(8 citation statements)

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“…from cattle have been molecularly characterised at the genes for 18S rRNA (18S rRNA), 28S rRNA (28S rRNA) and cytochrome c oxidase subunit I (cox1) and at nuclear rDNA internal transcribed spacer 1 (ITS1) [8,10,11,15]. Databases contain many 18S rRNA gene sequences of the genus Sarcocystis used for species identification [16][17][18]. However, studies have revealed that cox1 is the preferable target to differentiate taxonomically related Sarcocystis spp.…”

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confidence: 99%

Molecular identification of four Sarcocystis species in cattle from Lithuania, including S. hominis, and development of a rapid molecular detection method

et al. 2020

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Background Six 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. Methods The 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. Results Under 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%). Conclusions Although 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.

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confidence: 99%

Molecular identification of four Sarcocystis species in cattle from Lithuania, including S. hominis, and development of a rapid molecular detection method

et al. 2020

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“…The sarcocysts of S. matsuoae possessed slender finger-like protrusions (Fig. 2f) similar to those of S. entzerothi (Prakas et al, 2017). The size of their sarcocysts were also nearly the same, being 1115 × 112 μm in S. matsuoae (Table 3) and 1139 × 108 μm in S. entzerothi (Prakas et al, 2017).…”

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confidence: 82%

“…Hence, this cyst type may not be as prevalent as suggested by its repeated reporting, and the sarcocysts examined by LM or SEM in some of these studies might belong to a different species. The finger-like cyst-wall protrusions of the single cyst depicted in the TEM micrographs in these papers seem to have a broad round base and a thinner, more flattened distal end, causing the tip of the protrusions to bend over, which are features known from sarcocysts of S. entzerothi (Prakas et al, 2017), Sarcocystis bovifelis , Sarcocystis bovini and Sarcocystis sinensis (Gjerde, 2016a), whereas sarcocysts of S. tarandi have erect columnar/cylindrical protrusions (Gjerde, 1985). Moreover, since S. matsuoae was placed as a sister species to S. entzerothi , it seems likely that the sarcocyst depicted in the abovementioned TEM micrographs belong to S. matsuoae .…”

Section: Discussionmentioning

confidence: 90%

“…1, Fig. S2), sequences of type 3 were clearly separated from those of S. entzerothi from roe deer and farmed sika deer in Lithuania (Prakas et al, 2017; Rudaitytė-Lukošienė et al, 2018). Likewise, the cox1 and 18S rDNA sequences of type 3 shared an identity of only 94.3–94.4% and 96.9–97.6% respectively, with those of S. entzerothi .…”

Section: Discussionmentioning

confidence: 94%

“…The genus Sarcocystis comprises intracellular protozoan parasites that require two hosts, usually in a prey-predator relationship, to maintain their life cycle. More than 200 species in various mammals, birds and reptiles have been reported as valid based on their definitive or intermediate hosts, cross transmission studies, sarcocyst morphology and/or comparative molecular studies (Prakas et al, 2016, 2017; Gjerde, 2016b; Gjerde et al, 2017a, b, 2018). Prior to the molecular era, sarcocysts in intermediate hosts were assigned to species mainly on the basis of their morphology as seen in wet mounts or histological sections by light microscopy (LM) or in ultrathin sections by transmission electron microscopy (TEM).…”

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confidence: 99%

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Morphological and molecular characteristics of seven Sarcocystis species from sika deer (Cervus nippon centralis) in Japan, including three new species

Abe

Matsuo

Moribe

et al. 2019

International Journal for Parasitology: Parasites and Wildlife

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Samples of diaphragm were collected from 53 sika deer from Gifu Prefecture, Japan; 220 sarcocysts were isolated, examined in wet mounts and classified according to their cyst wall protrusions. The sarcocysts were then examined molecularly in order to assign them to different species. All but 11 of the 220 sarcocysts were initially identified by means of a multiplex PCR assay targeting cox1 of five species, whereas the remaining 11 sarcocysts were identified by standard PCR and sequencing. DNA from selected sarcocysts was used for PCR amplification and sequencing of cox1 (59 sequences) and 18S rDNA (23 sequences). The 220 sarcocysts comprised seven major cox1 sequence types or species. Types 4 and 7 were assigned to the known species Sarcocystis pilosa and Sarcocystis ovalis, whereas types 1, 3 and 5 were considered to represent three new species, for which the names Sarcocystis japonica, Sarcocystis matsuoae and Sarcocystis gjerdei have been proposed. Types 2 and 6 were most similar to Sarcocystis tarandi and Sarcocystis taeniata, respectively, but could not be unequivocally assigned to these species. Sarcocysts belonging to S. japonica were macroscopic with fairly thick finger-like protrusions, whereas most sarcocysts of the six other species were microscopic. Sarcocysts of S. cf. tarandi and S. matsuoae were spindle-shaped and possessed thin finger-like cyst-wall protrusions. Sarcocysts of S. pilosa and S. gjerdei had similar hair-like protrusions, whereas those of S. cf. taeniata had a smooth surface. Sarcocysts of S. japonica, S. pilosa, S. cf. tarandi, S. gjerdei, S. matsuoae, S. cf. taeniata and S. ovalis were found in 50 (94.3%), 29 (54.7%), 22 (41.5%), 10 (18.9%), 8 (15.1%), 6 (11.3%) and 1 (1.9%) of the 53 sika deer examined, respectively. An improved multiplex PCR assay targeting cox1 was developed, through which the seven Sarcocystis spp. found in the present study could be identified.

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Morphological and molecular characterization of four Sarcocystis spp., including Sarcocystis linearis n. sp., from roe deer (Capreolus capreolus) in Italy

et al. 2017

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Fresh (frozen/thawed) muscle samples from four 2-12-year-old roe deer (Capreolus capreolus) from the Sondrio province in north-eastern Italy were examined under a dissecting microscope, and about 180 sarcocysts were isolated and identified to morphological type in wet mounts by light microscopy (LM). Seventy-seven of these sarcocysts were subsequently examined by molecular methods, comprising polymerase chain reaction (PCR) amplification and sequencing of the partial cytochrome c oxidase subunit I gene (cox1) of all isolates, as well as PCR amplification, cloning and sequencing of the complete18S ribosomal RNA (rRNA) gene of two isolates of each species found. By LM, three major sarcocyst types were recognised: spindle-shaped sarcocysts, 0.5-3 mm long, either with no clearly recognisable protrusions (thin-walled) or with finger-like protrusions (thick-walled); and slender, thread-like sarcocysts, 2-3 mm long, with hair-like protrusions. Sequencing of cox1 revealed that the sarcocysts belonged to four different species. Those with no visible protrusions either belonged to Sarcocystis gracilis (n = 24) or to a Sarcocystis taeniata-like species (n = 19), whereas those with finger- and hair-like protrusions belonged to Sarcocystis silva (n = 27) and Sarcocystis capreolicanis (n = 7), respectively. The 19 cox1 sequences of the S. taeniata-like species, comprising five haplotypes, differed from each other at 0-16 of 1038 nucleotide positions (98.5-100% identity). They differed from 25 previous cox1 sequences of S. taeniata from moose and sika deer (with 98.0-100% intraspecific identity), at 33-43 nucleotide positions (95.9-96.8% interspecific identity), and there were 20 fixed nucleotide differences between the two populations. In the phylogenetic analysis based on cox1 sequences, the two populations formed two separate monophyletic clusters. The S. taeniata-like species in roe deer was therefore considered to represent a separate species, which was named Sarcocystis linearis n. sp. At the 18S rRNA gene, however, the two species could not be clearly separated from each other. Thus, there was considerable intraspecific sequence variation in the 18S rRNA gene of S. linearis (98.1-99.9% identity between 24 sequences), which was similar both in magnitude and nature to the variation previously found in this gene of S. taeniata. The new 18S rRNA gene sequences of S. linearis shared an identity of 97.9-99.6% with those of S. taeniata (overlap between intra- and interspecific identity), and in the phylogenetic tree, sequences of the two species were interspersed. By scanning electron microscopy (SEM), the sarcocysts of S. linearis were found to possess regularly spaced, thin and narrow ribbon-like cyst wall protrusions (about 2.8-3.2 μm long, 0.3-0.4 μm wide and about 0.02-0.03 μm thick), terminating in a plate-like structure of the same thickness but with an elliptic outline (about 0.3-0.4 μm wide and 0.7-0.9 μm long). The terminal plates were connected in the middle with the band-like portion of the protrusions like the boar...

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Sarcocystis entzerothi n. sp. from the European roe deer (Capreolus capreolus)

Cited by 22 publications

References 44 publications

Molecular identification of four Sarcocystis species in cattle from Lithuania, including S. hominis, and development of a rapid molecular detection method

Molecular identification of four Sarcocystis species in cattle from Lithuania, including S. hominis, and development of a rapid molecular detection method

Morphological and molecular characteristics of seven Sarcocystis species from sika deer (Cervus nippon centralis) in Japan, including three new species

Morphological and molecular characterization of four Sarcocystis spp., including Sarcocystis linearis n. sp., from roe deer (Capreolus capreolus) in Italy

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