This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Until recently, the detection, identification and assignment of Entamoeba organisms to species relied mainly on morphology and the host in which parasites were identified (Stensvold et al., 2010;Stensvold et al., 2011). However, morphology is not a reliable tool for delimiting Entamoeba species as cyst morphology varies substantially within as well as between uninucleated cyst-producing species from different ruminant hosts (Noble and Noble, 1952;Pillai and Kain, 1999;Stensvold et al., 2010). The use of molecular tools is therefore essential to resolve the identification, taxonomy, epidemiology and clinical significance of Entamoeba species without reliance on parasite cultures or experimental infections (Stensvold et al., 2011; Jacob et al., 2016).Rangeland goats are an introduced animal species in Australia. They can be legally trapped and reared by licensed operators (goat depots) for the domestic and export meat markets, which was worth approximately $AUS242 million in 2014 (MLA, 2015). Few 4 studies have conducted genetic characterisation of Entamoeba species from ruminants (Stensvold et al., 2010; Jacob et al., 2016), which is important for understanding their evolutionary and taxonomic relationships. In the present study, uninucleate Entamoeba cysts were identified in the faeces of Rangeland goats in Western Australia by microscopy and were characterised at the 18S ribosomal RNA (rRNA) and actin loci.
Materials and methodsFaecal samples were collected by rectal palpation from 125 male AustralianRangeland goats (Capra hircus) on arrival at a commercial goat depot near Geraldton, Western Australia, after capture and transport from a sheep and cattle extensive rangeland grazing property, North Wooramel station, located 78 km east of Denham and 113 km south east of Carnarvon in the Gascoyne region of Western Australia. On arrival, the goats weighed on average 30.7 ± 0.3 kg (±SEM), and the estimated age of the goats, based on dentition, was between 9 -12 months. Faecal samples were immediately placed on ice until transported to the lab, where microscopy work was performed on the same day of collection and then stored in the refrigerator (4.0°C) until DNA extraction was performed. All sample collection methods used were approved by the Murdoch University Animal Ethics Committee (approval number R2617/13).Direct microscopic examination of faecal suspensions in saline and wet mounted with 0.9% saline and Lugol's iodine was conducted. Entamoeba cysts were concentrated using zinc-sulfate gradient floatation (Faust's method) (Ramos et al., 2005) and observed ...