Abstract

Red deer (Cervus elaphus) are hosts of liver fluke (Fasciola hepatica); yet, prevalence is rarely quantified in wild populations. Testing fresh samples from remote regions by faecal examination (FE) can be logistically challenging; hence, we appraise frozen storage and the use of a coproantigen ELISA (cELISA) for F. hepatica surveillance. We also present cELISA surveillance data for red deer from the Highlands of Scotland. Diagnoses in faecal samples (207 frozen, 146 fresh) were compared using a cELISA and by FE. For each storage method (frozen or fresh), agreement between the two diagnostics was estimated at individual and population levels, where population prevalence was stratified into cohorts (e.g., by sampling location). To approximate sensitivity and specificity, 65 post-slaughter whole liver examinations were used as a reference. At the individual level, FE and cELISA diagnoses agreed moderately (κfrozen = 0.46; κfresh = 0.51), a likely reflection of their underlying principles. At the population level, FE and cELISA cohort prevalence correlated strongly (Pearson’s R = 0.89, p < 0.0001), reflecting good agreement on relative differences between cohort prevalence. In frozen samples, prevalence by cELISA exceeded FE overall (42.8% vs. 25.8%) and in 9/12 cohorts, alluding to differences in sensitivity; though, in fresh samples, no significant difference was found. In 959 deer tested by cELISA across the Scottish Highlands, infection prevalence ranged from 9.6% to 53% by sampling location. We highlight two key advantages of cELISA over FE: i) the ability to store samples long term (frozen) without apparent loss in diagnostic power; and ii) reduced labour and the ability to process large batches. Further evaluation of cELISA sensitivity in red deer, where a range of fluke burdens can be obtained, is desirable. In the interim, the cELISA is a practicable diagnostic for F. hepatica surveillance in red deer, and its application here has revealed considerable geographic, temporal, sex and age related differences in F. hepatica prevalence in wild Scottish Highland red deer.

Highlights

  • Red deer (Cervus elaphus) are a recognised host of liver fluke (Fasciola hepatica) throughout Europe [1,2,3,4,5]

  • In 146 fresh faecal samples collected during 2013–14, prevalence of F. hepatica estimated by faecal examination (FE) (12.3%) and coproantigen enzyme-linked immunosorbent assay (cELISA) (9.6%) did not differ significantly (McNemar’s χ2 = 0.64, df = 1, p-value = 0.42) (S1 Table); whereas in 159 frozen samples collected during 2012–13, prevalence estimated by cELISA (42.8%) was significantly greater than FE (25.8%) (McNemar’s χ2 = 16.5, df = 1, p-value 10 per cohort), prevalence estimated by cELISA in frozen samples exceeded FE in 4/6 cohorts; whereas in fresh samples, prevalence estimated by cELISA only exceeded FE in 1/7 cohorts

  • Sex and estate (S2 Table; where n > 10 per cohort), prevalence in frozen samples estimated by cELISA exceeded FE in 9/12 cohorts, and estimates of prevalence by cELISA and FE were strongly correlated (Pearson’s R = 0.89, p < 0.0001) (Fig 1); whereas in fresh samples, prevalence estimated by cELISA only exceeded FE in 2/5 and the correlation between the cELISA and FE was not significant (Pearson’s R = 0.85, p < 0.067)

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Summary

Introduction

Red deer (Cervus elaphus) are a recognised host of liver fluke (Fasciola hepatica) throughout Europe [1,2,3,4,5]. In contrast to domestic ruminants, in which disease monitoring is routine, apart from a handful of species, (namely, coypu (Myocastor coypus) [6,7], European hare (Lepus europaeus) [8,9], European rabbit (Oryctolagus cuniculus) [8], wild boar (Sus scrofa) [10] and roe deer (Capreolus capreolus) [11]), data concerning F. hepatica in wildlife is limited to incidental observations, which is the case for red deer in the Scottish Highlands [3]. F. hepatica prevalence of 11% and 40–90% egg viability, implicate wild boar (S. scrofa) as a likely reservoir to cattle in NW Spain [10]

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