Research Brief First molecular identification of Sarcocystis miescheriana (Protozoa, Apicomplexa) from wild boar (Sus scrofa) in Iran Eshrat Beigom Kia a , Hossein Mirhendi a , Mostafa Rezaeian a , Farzaneh Zahabiun a , Mitra Sharbatkhori b,⇑ a Department of Medical Parasitology & Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran b Department of Medical Parasitology & Mycology, School of Medicine, Golestan University of Medical Sciences, Gorgan, Iran article info Article history: Received 24 July 2010 Received in revised form 13 November 2010 Accepted 16 November 2010 Available online 21 November 2010 Keywords: Sarcocystis miescheriana Protozoa Apicomplexa Wild boar Iran abstract Sarcocystis isolate obtained from the thigh muscle of a wild boar (Sus scrofa), captured from Gilan Province, northern Iran, was subjected to molecular analysis. Genomic DNA was obtained using a DNA extraction tissue kit and Polymerase chain reaction (PCR) for amplification of the 18S ribosomal DNA region yielded an 842 bp DNA band on agarose gel. Analysis of DNA sequencing by BLAST confirmed the isolate as Sarcocystis miescheriana and the sequence was deposited in GenBank by Accession No. GU395554. This is the first molecular identification of an isolate of S. miescheriana in Iran. Ó 2010 Elsevier Inc. All rights reserved. 1. Introduction Sarcocystis is a genus of cyst-forming coccidia belonging to the phylum Apicomplexa. This genus is comprised of more than 200 species (Frenkel and Smith, 2003). The organism has an obligatory heteroxenous life cycle, with a sexual stage in enteroepithelial cells of the definitive host, and asexual generation in the tissues of the intermediate host (Dubey et al., 1989). Usually herbivores and omnivores serve as intermediate hosts while omnivores and carni- vores are definitive hosts (Lindsay et al. 1995). Intermediate and definitive hosts can harbor various Sarcocystis spp (Mehlhorn and Heydorn, 1978). Species in the family Suidae generally harbor tissue cysts of two species of Sarcocystis, namely Sarcocystis mie- scheriana (Kühn, 1865) Labbé, 1899 (synonym: Sarcocystis suicanis) (Heydorn et al., 1975; Erber, 1977; Mehlhorn and Heydorn, 1978) and S. suihominis (Tadros & Laarman, 1976) (Heydorn, 1977). The validity of third species, Sarcocystis porcifelis (Dubey, 1976) is still unclear (Dubey et al., 1989). Species identification in Sarcocystis is usually performed by morphological characterization of sarcocyst especially cyst wall structure and sporocysts under light or transmission and scanning electron microscopy. Since the appearance may change depending on the location and developmental stage of sarcocyst and other conditions of parasitized cell, molecular studies have been sug- gested to confirm morphological species identification (Levine, 1986; Dahlgren and Gjerde, 2007). Gene sequence data analysis is also very useful tool to clarify whether morphologically similar sarcocysts in intermediate hosts are the same or different species (Yang et al., 2001). In Iran, some species of livestock are commonly parasitized by species of Sarcocystis. There have been several studies on preva- lence of infection in cattle and sheep from different parts of the country, but a few have identified the species involved (Naghibi et al., 2002; Dalimi et al., 2008). In the present study, the first molecular identification of S. miescheriana from a wild boar in northern Iran is documented. 2. Materials and methods The source of parasite was a portion of a frozen thigh muscle of a wild boar hunted in April 2007 from Javaher Dasht Forest, Siah- kal, Gilan Province, a temperate area in northern Iran, bordering the Caspian Sea. A portion of the muscle tissue was prepared for direct examina- tion by light microscope. Gross inspection of unstained tissue smears revealed the presence of Sarcocystis cysts. A piece of muscle was preserved in 10% formalin and after tissue processing using conventional histological methods, and 5 lm sections were pre- pared and stained with H&E (haematoxylin-eosin). For molecular identification, small pieces of muscles were pre- served in 80% ethanol alcohol until further use. Total genomic DNA was extracted from a small piece of muscle stored in ethanol, employing DNeasy blood & tissue (Qiagen, Hilden, Germany) 0014-4894/$ - see front matter Ó 2010 Elsevier Inc. All rights reserved. doi:10.1016/j.exppara.2010.11.007 ⇑ Corresponding author. Fax: +98 171 4440225. E-mail address: mitra.sharbatkhori@gmail.com (M. Sharbatkhori). Experimental Parasitology 127 (2011) 724–726 Contents lists available at ScienceDirect Experimental Parasitology journal homepage: www.elsevier.com/locate/yexpr