Sarcocystis spp. infection in South American deer huemul ( Hippocamelus bisulcus ) and pudu ( Pudu puda ) from Patagonia

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PROTOZOOLOGY - SHORT COMMUNICATION

Sarcocystis spp. infection in South American deer huemul (Hippocamelus bisulcus) and pudu (Pudu puda) from Patagonian National Parks, Argentina Elizabeth Chang Reissig 1,2

&

Elisa Helman 2,3 & Gastón Moré 2,3

Received: 27 March 2020 / Accepted: 13 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020

Abstract Sarcocystis spp. are intracellular protozoan parasites with heteroxenous life cycles. This study described Sarcocystis spp. infection in adult South American native deer huemul (Hippocamelus bisulcus) and pudu (Pudu puda). Heart, diaphragm, tongue, and skeletal muscle samples were collected from 5 huemuls and 2 pudus, found dead in National Parks. Direct microscopic examination, transmission electron microscopy, PCR, and sequencing were performed. Sarcocystis spp. microscopic thinwalled cysts were identified in 3 huemuls and 1 pudu. Several cysts from 1 huemul and 1 pudu were observed by TEM; ultrastructure was similar to previously reported as cyst wall type 17 and types 2 and 8, respectively. Fragments of the 18S rRNA and cytochrome c oxidase subunit I (cox1) genes were amplified and sequenced from 3 individual cysts from 2 huemuls and 2 cysts from the pudu. The sequences from huemuls showed a high identity among them (> 99%) at both amplified targets. The highest identities were > 99.7% at 18S rRNA and 93% at cox1 with S. tarandivulpes sequences. The 18S rRNA gene sequences from pudus showed an identity > 99.7% with Sarcocystis sp., S. taeniata, and S. linearis sequences, while the cox1 sequences were different, one showing 99.42% identity with S. venatoria and the other 98.22% with S. linearis. A single species, similar to S. tarandivulpes, was identified in all huemul samples while 2 molecularly different Sarcocystis spp. were found in 1 pudu with high similarities to either S. venatoria or to S. linearis, S. taeniata-like, and S. morae. Based on the cox1 sequence identities, at least the Sarcocystis sp. in huemuls might represent a new species, primarily occurring in this host. Additional sarcocyst isolates from both hosts need to be examined molecularly in order to firmly establish whether these species are indeed native to huemuls and/or pudus or are derived from introduced deer species. Keywords Sarcocystis . Ultrastructure . PCR-sequencing . Native cervids/wild ungulates Section Editor: Daniel K Howe Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00436-020-06889-9) contains supplementary material, which is available to authorized users. * Gastón Moré [email protected] 1

Natural Resources Department, Institute of Forest and Agriculture Research Bariloche (IFAB, CONICET-INTA), Agricultural Experimental Station of the National Institute of Agriculture and Technology (EEA-INTA Bariloche), Modesta Victoria 4455, (8400) San Carlos de Bariloche, Río Negro, Argentina

2

Argentinean National Council of Scientific and Technological Research (CONICET), Godoy Cruz 2290, CABA (C1425FQB) Buenos