2017)Morphological and molecular characterization of three Eimeria species from captured rangeland goats in Western Australia. Veterinary Parasitology: Regional Studies and Reports, 9. pp. 75-83. ABSTRACT Faecal shedding of Eimeria by captured rangeland goats (Capra hircus) was investigated using a longitudinal observational study. Faecal samples were collected from 125 male goats on four occasions. The first sampling occurred following capture and transport, immediately after arrival at a commercial goat depot (feedlot) in Western Australia, with subsequent 3 sample collections occurring at one month intervals thereafter. Goats were composite breed and aged approximately 9-12 months on arrival at the feedlot. Prevalence and shedding intensity (faecal oocyst concentration) for Eimeria were determined using qPCR. Species were identified from individual oocysts (isolated using micromanipulation) using molecular analysis at two loci, specifically 18S rRNA and mitochondrial cytochrome oxidase gene (COI), and confirmed by microscopy. Longitudinal prevalence (animals positive at least once) for Eimeria spp. by qPCR was 90.4%, with 60% goats shedding Eimeria spp. on more than one occasion. Point prevalence (prevalence at a single sampling occasion) ranged from 2.4% (fourth sampling) to 70.4% (second sampling). Three species were identified at the 18S rRNA locus and confirmed by microscopy: E. christenseni (longitudinal prevalence for single infection 34.4%), E. hirci (17.6%) and E. arloingi (8.8%) over the four sample collections. Mixed infections were identified in 56.8% goats (longitudinal prevalence). 18S rRNA sequences from E. christenseni and E. hirci and were 100% homologous with ovine E. ahsata and E. crandallis respectively, and E. arloingi was 100% similar to caprine E. arloingi. At the COI locus, E. christenseni, E. hirci and E. arloingi grouped separately, and were closely related to ovine E. ahsata, with genetic similarities of 96.5%, 92.6% and 91.4% respectively. This is the first report for molecular characteristics of caprine-derived Eimeria spp. using a combination of 18S rRNA and COI. Molecular techniques can be used to identify Eimeria ACCEPTED MANUSCRIPT A C C E P T E D M A N U S C R I P T spp. in goat faecal samples, specifically through characterization at 18S locus and other gene loci when used in parallel. Molecular techniques offer some advantages over microscopy for identification of Eimeria species, particularly with respect to precision.

No abstract

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. A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT A C C E P T E D M A N U S C R I P T1 ABSTRACT Faecal shedding of Cryptosporidium and Giardia by captured rangeland goats was investigated using a longitudinal study with four faecal samples collected from 125 male goats once monthly for four months, commencing immediately after capture and transport to a commercial goat depot (feedlot). Goats were composite breed and aged approximately 9-12 months on arrival. Faecal samples were screened for Cryptosporidium and Giardia presence and concentration using quantitative PCR and sequencing at the 18S ribosomal RNA locus (Cryptosporidium), and glutamate dehydrogenase and β-giardin loci (Giardia). Longitudinal prevalence for Cryptosporidium was 27.2% (point prevalence range 3-14%) with 3 species identified: C. xiaoi (longitudinal prevalence 13.6%), C. ubiquitum (6.4%) and C. parvum (3.2%). Sub-typing at the gp60 locus identified C. ubiquitum XIIa, C. parvum IIaA17G2R1 and C. parvum IIaA17G4R1. This is the first report of the zoonotic C. parvum subtype IIaA17G4R1 in goats. The pattern of genotypes shed in faeces changed over the duration of study with C. ubiquitum identified only at the first and second samplings, and C. parvum identified only at the fourth sampling. Longitudinal prevalence for Giardia duodenalis was 29.6% (point prevalence range 4-12%) with all positives sub-typed as assemblage E. Only 2/125 goats were identified to be shedding Cryptosporidium or Giardia on more than one occasion. This is the first report of Cryptosporidium and Giardia genotypes in captured rangeland goats. Faecal shedding of zoonotic Cryptosporidium spp. and potentially zoonotic G. duodenalis has implications for food safety and effluent management

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 ...

Caprine arthritis encephalitis (CAE) is a chronic debilitating disease of goats caused by a lentivirus responsible for economic losses as a result of a drop in milk production and weight loss. The objective of the study was to determine if indigenous goats from five different regions in the Sultanate of Oman exhibit serological evidence of exposure to CAEV using a competitive-inhibition ELISA technique. Blood samples were collected from slaughtered goats (N=1,110) and from the National Serum Bank (n=528). In total, 83 (5.1%) of screened samples were classed as seropositive. The results provide the first serological evidence for the presence of CAEV in Oman.