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MODIFIED DIGESTION TECHNIQUE FOR DETECTION OF SARCOCYSTIS BRADYZOITES.

Sarcocystis spp. is a zoonotic intracellular protozoan parasite with an obligatory two-host life cycle causing a disease namely sarcosporidiosis or sarcocystosis (Fazly Ann et al. 2013). According to Hamidinejat et al. (2010) the gold standard in detecting sarcocystis is a digestion technique. It is a commonly apply technique and considered one of the most sensitive methods to detect the presence of sarcocystis bradyzoites in muscular tissues (Dubey et al. 1989). The aim of this study is to detect the presence of sarcocystis bradyzoites using modified digestion technique. Fifty grams of each skeletal muscle samples were minced and homogenized using blender containing 100 ml distilled water. The homogenized samples were then settled down for 5 minutes in a 250 ml beaker. After removal of the supernatant, the remaining 50 ml sediment was digested with pepsin (Sigma®, USA) solution and 1.5% hydrochloric acid (HmbG®, Germany). The mixtures were then incubated at 30°C in the waterbath (Memmert W350, Germany) for 12 hours. The digested samples were sieved through a nylon-meshed tea strainer and centrifuged at 1500 rpm for 5 minutes (Eppendorf Centrifuge 5804R, Germany). After discarding the supernatant, a drop of sediment was stained with Giemsa (Sigma®, USA) and examined under the light microscope (Leica DME, USA) at x400 magnification for detection of sarcocystis bradyzoites. A few digestion techniques have been published using different types of enzyme as digestion medium (Dubey et al. 1989; Beyazit et al. 2007; Rahdar and Salehi 2011; Obijiaku et al. 2013). Bradyzoites are released from the sarcocyst by incubation in the digestion medium containing either trypsin, or pepsin and hydrochloric acid (Huong 1999). In this technique, the sarcocyst wall was dissolved by peptic digestion. The released bradyzoites survive appear crescent or banana-shape under phase contrast microscope. The length of bradyzoites may differ remarkably from 3-5µm up to 18µm depending on the species of sarcocystis. The shape may also vary according to their density within the cysts or the method of preparation. As a recommendation, this developed modified digestion techniques should be taken into consideration to be used as an alternative for detection of the presence of sarcocystis bradyzoites.

Fazly Ann, Z., N. A. Muhamad Syamsul Naim, W.O.B. Wan Normaziah, S. Geethamalar, M.I. Iswadi and M. Ramlan 

Veterinary Research Institute, 59 Jalan Sultan Azlan Shah, 31400, Ipoh, Perak, Malaysia

Corresponding Author: Fazly Ann Zainalabidin

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Fazly Ann, Z., N. A. Muhamad Syamsul Naim, W.O.B. Wan Normaziah, S. Geethamalar, M.I. Iswadi and M. Ramlan (2014) Modified Digestion Technique for Detection of Sarcocystis Bradyzoites. 1st ASEAN Regional Conference on Animal Production (ARCAP) & 35th Malaysian Society of Animal Production (MSAP) Annual Conference “Bridging Technology Gap for ASEAN Animal Production”, 4th-6th June 2014, Riverside Majestic Hotel Kuching, Sarawak, Malaysia, pp.189-190


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