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. 2023 Jan 2:20:39-45.
doi: 10.1016/j.ijppaw.2022.12.007. eCollection 2023 Apr.

Identification of a new Sarcocystis sp. in marsh deer (Blastocerus dichotomus) from wetlands of Argentina

Yanina Berra  1   2 , Gastón Moré  3   2 , Elisa Helman  3   2 , Hernan D Argibay  4 , M Marcela Orozco  2   5
Affiliations

Identification of a new Sarcocystis sp. in marsh deer (Blastocerus dichotomus) from wetlands of Argentina

Yanina Berra et al. Int J Parasitol Parasites Wildl. .

Abstract

The marsh deer (Blastocerus dichotomus) is the largest South American native deer species and is listed as "Vulnerable" by IUCN due to the population reduction. As part of a conservation and disease surveillance program, muscle samples from 14 marsh deer found dead in 2016 and 2017 in northeast Argentina were obtained at necropsy. Samples from each animal were processed as pooled muscles (heart, diaphragm, tongue and hindlimb) by homogenization and direct microscopical observation to detect intracellular Sarcocystis spp. cysts. Sarcocysts were observed in six samples, and several cysts recovered from two samples were processed by transmission electron microscopy. The cysts were thin-walled and showed a cyst-wall ultrastructure with ribbon-like protrusions similar to other species using cervids as intermediate host and canids as definitive hosts. Genomic DNA from individual sarcocysts from three marsh deer were successfully amplified by PCR of 18S rRNA and COI gene fragments and further sequenced. Sequence comparison revealed a 99.3-100% identity among them and only 93.7-96.6% and 88.8-89.7% identity at 18S rRNA and COI markers, respectively, with other Sarcocystis spp. Despite morphological similarities, the high sequence divergence at 18S rRNA and COI fragments allowed the assumption that Sarcocystis sp. from marsh deer is a different species from others using cervids as intermediate hosts. Therefore, we propose the name Sarcocystis blastoceris n. sp. for the species infecting marsh deer.

Keywords: Blastocerus dichotomus; Marsh deer; PCR-Sequencing; Sarcocystis; Ultrastructure.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Image 1
Graphical abstract
Fig. 1
Fig. 1
A and B: Microscopical sarcocyst detected in marsh deer muscles. A: Optical microscopy image from a complete cyst. B: higher magnification of a portion from the cyst displayed in A. Note the thin cyst wall without apparent protrusions.
Fig. 2
Fig. 2
Transmission electron microscopy (TEM) image of the cyst wall from a microscopical sarcocyst in a marsh deer muscle (17-12CDP). Note the primary cyst wall (Pcw) from which arise bent ribbon-like protrusions folded over the cyst surface. The ground substance layer (gs) showed no granules nor microtubules. Electron lucid amylopectin granules (am) appear irregularly distributed within vacuolated bradyzoites (vb). Both, bradyzoites and host muscle cell are decomposed.
Fig. 3
Fig. 3
Neighbor-Joining consensus phylogenetic tree. Phylogenetic tree based on an alignment of 41 Sarcocystis spp. 18S rRNA sequences performed with GENEIOUS software (Version R9), using a Tamura-Nei genetic distance model. The three sequences from marsh deer obtained in the present study are in bold. Branch consensus support is expressed as % from 1000 bootstraps. Sequence M97703 from T. gondii used as outgroup.
Fig. 4
Fig. 4
Neighbor-Joining consensus phylogenetic tree. Phylogenetic tree based on an alignment of 40 Sarcocystis spp. COI sequences performed with GENEIOUS software (Version R9), using a Tamura-Nei genetic distance model and no outgroup. The three sequences from marsh deer (one from each animal) are in bold. Branch consensus support is expressed as % from 1000 bootstraps. Sequence JX473257 from T. gondii used as outgroup.
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