The Eimeria species are highly pathogenic parasites of @entity1573 . Research aimed at reducing their impact is hindered by a lack of non-subjective, quantitative, tools to measure parasite replication in the host. The time-consuming, and often time-sensitive, nature of existing approaches precludes their use in large-scale genetic, epidemiological, and evolutionary analyses. We have used quantitative real-time PCR (qPCR) to accurately quantify @entity12917 in @entity1573 tissue and shown this to be more efficient and sensitive than traditional methodologies. We tested four @entity1573 -specific reference qPCR assays and found beta-actin (actb) to be optimal for sample normalisation. In an experimental setting, @entity1573 were inoculated with 500, 1500, or 4500 @entity12917 oocysts and parasite replication and the impact of @entity281 measured by i) qPCR analysis of DNA extracted from caecal tissues collected at five and eight days post- @entity281 (dpi), ii) faecal oocyst counts (FOCs) on samples taken from six to eight dpi, and iii) lesion scoring on caeca collected post-mortem at five and eight dpi. Quantitative real-time PCR test results indicated a significant dose-dependent increase in parasite numbers among study groups for samples collected five dpi (i.e., prior to gametogony) (R(2)=0.994) (p<0.002) but not in those from day eight (after most oocyst shedding) (R(2)=0.006) (p>0.379). A strong dose-dependent increase in parasite replication and severity of @entity281 was also revealed by FOC (R(2)=0.997) and lesion scoring. Importantly, qPCR offers substantial improvements for animal welfare via improved statistical power and reduced group sizes in experimental studies. The described qPCR method overcomes subjective limitations of coproscopic quantification, allows reproducible medium- to high-throughput examination of tissues, faeces, and oocysts, and is a valuable tool for determining the impact of @entity281 in both experimental and field settings.

Quantitative real-time PCR (qPCR) for XXXX replication--Implications for experimental refinement and animal welfare.

@entity1573 :: ('9031', 'Species') :: ['chickens', 'chicken'] @entity12917 :: ('5802', 'Species') :: ['Eimeria tenella', 'E. tenella'] @entity281 :: ('MESH:D007239', 'Disease') :: ['Eimeria infections', 'infection']
@entity12917 :: (5802,Species) :: ['Eimeria tenella']
