Development of a quantitative PCR (qPCR) for Giardia and analysis of the prevalence, cyst shedding and genotypes of Giardia present in sheep across four states in Australia

Abstract

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Greater intensity and frequency of Cryptosporidium and Giardia oocyst shedding beyond the neonatal period is associated with reductions in growth, carcase weight and dressing efficiency in sheep

“…Giardia spp. were identified in 20.2% of faecal samples (Table 3), with prevalence and faecal oocyst concentration previously reported in greater detail (Yang et al, 2014b Cryptosporidium spp. shedding at pre-slaughter was associated with about 1.2% lower dressing %, within both high shedding (P<0.01) and low shedding (P<0.01) lambs compared to non-shedding lambs (Table 4).…”

“…The mean RSQ (R squared) was 0.98 and the % Relative Standard Deviation (RDS) = 5.5% (Yang et al, 2014b).…”

“…The selection of animals included in this experiment and sample collection methods have been previously described (Yang et al, 2014a;2014b). In brief, lambs from eight farms were included in this study (Table 1).…”

“…All Cryptosporidium positives were screened using a C. parvum and C. hominis specific qPCR at a unique Cryptosporidium specific gene (Clec) that codes for a novel mucin-like glycoprotein that contains a C-type lectin domain (CTLD) previously described (Morgan et al, 1996;Yang et al, 2009;Bhalchandra et al, 2013). Cryptosporidium and Giardia oocysts per gram of faeces (g -1 ) were measured by qPCR as previously described (Yang et al, 2014a;2014b). PCR contamination controls were used including negative controls and separation of preparation and amplification areas.…”

“…All samples were screened at the actin locus for Cryptosporidium and Giardia at the glutamate dehydrogenase (gdh) locus using quantitative PCR (qPCR) previously described (Yang et al, 2014a;2014b). An internal amplification control (IAC) which consisted of a fragment of a coding region from Jembrana Disease Virus cloned into a pGEM-T vector (Promega, USA), was used as previously described (Yang et al, 2013).…”