Abstract
The Faecal Egg Count Reduction Test (FECRT) is the most widely used field-based method for estimating anthelmintic efficacy and as an indicator of the presence of anthelmintic resistant nematodes in cattle, despite never having been validated against the gold standard of controlled slaughter studies. The objectives of this study were to assess the normality of cattle faecal egg count (FEC) data and their transformed versions, since confidence intervals used to aid the interpretation of the FECRT, are derived from data assumed to be normally distributed, and violation of this assumption could potentially lead to the misclassification of anthelmintic efficacy. Further, probability distributions and associated parameters were evaluated to determine those most appropriate for representing cattle FEC data, which could be used to estimate percentage reductions and confidence limits. FEC data were analysed from 2175 cattle on 52 farms using a McMaster method at two different diagnostic sensitivities (30 and 15 eggs per gram (epg)) and a sensitive centrifugal flotation technique (SCFT) with a sensitivity of 1 epg. FEC data obtained from all egg count methods were found to be non-normal even upon transformation; therefore, it would be recommended that confidence or credible intervals be generated using either a Bootstrapping or Bayesian approach, respectively, since analyses using these frameworks do not necessarily require the assumption of normality. FEC data obtained using the SCFT method were best represented by distributions associated with the negative binomial and hence arithmetic means could be used in FECRT calculations.Where FEC data were obtained with less sensitive counting techniques (i.e. McMaster 30 or 15 epg), zero-inflated distributions and their associated central tendency were the most appropriate and would be recommended to use, i.e. the arithmetic group mean divided by the proportion of non-zero counts present; otherwise apparent anthelmintic efficacy could be misrepresented.
Highlights
For over 60 years the control of helminth parasites, due to their ever growing impact on animal health and welfare (Crofton, 1966; Vlassoff and McKenna, 1994; Corwin, 1997; Molento, 2009; Voort et al, 2013; Charlier et al, 2014), has increasingly relied on the use of anthelmintics
The true representation of resistance is difficult to assess mainly due to inconsistencies in treatment dose administrations, faecal sample collection and handling methods, faecal egg counting techniques used, associated experimental designs (Taylor, 2012) and the lack of robust methods for determining anthelmintic resistance under field conditions i.e. the lack of field data supported by controlled slaughter studies, or the availability of validated molecular and in-vitro methods for cattle nematodes
Faecal Egg Count Reduction Test (FECRT) Methods for calculating the FECRT involve determining the arithmetic group mean Faecal egg counts (FECs) on Day 0, and/or Day 14, and calculating the percentage reduction and the 95% confidence intervals for treated and untreated groups of animals
Summary
For over 60 years the control of helminth parasites, due to their ever growing impact on animal health and welfare (Crofton, 1966; Vlassoff and McKenna, 1994; Corwin, 1997; Molento, 2009; Voort et al, 2013; Charlier et al, 2014), has increasingly relied on the use of anthelmintics. The true representation of resistance is difficult to assess mainly due to inconsistencies in treatment dose administrations, faecal sample collection and handling methods, faecal egg counting techniques used, associated experimental designs (Taylor, 2012) and the lack of robust methods for determining anthelmintic resistance under field conditions i.e. the lack of field data supported by controlled slaughter studies, or the availability of validated molecular and in-vitro methods for cattle nematodes. The most common method used to investigate anthelmintic resistance is the Faecal Egg Count Reduction Test (FECRT) (Coles et al, 1992, 2006). This test has not been validated against slaughter studies and the European Medicines Agency (EMA) regards this test as an estimation of efficacy, and not confirmation of resistance (EMA, 2014). True resistance must be confirmed through laboratory slaughter studies, potentially supported by molecular level studies, or methods such as egg hatch tests (Vidyashankar et al, 2012)
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