Comparison of the Modified McMaster and Mini-FLOTAC methods for the enumeration of nematode eggs in egg spiked and naturally infected chicken excreta

Abstract

Excreta egg counting techniques are used for indirectly estimating the magnitude of gastrointestinal nematode infection in live animals. The aim of this study was to optimise laboratory and field sampling methods for routine monitoring of nematode infections in chickens by evaluating the sensitivity, accuracy, and precision of the Modified McMaster (MM) and Mini-FLOTAC (MF) methods using laying chicken excreta samples spiked with estimated true numbers of eggs (Experiment 1 = 5-1500 EPG (eggs/g); Experiment 2 = 5-500 EPG) without and with operator effects, respectively or using individual fresh excreta (n = 230) and fresh floor excreta (n = 42) from naturally infected free-range layer farms. The Coefficient of Variation (CV) was assessed within and between operators and the time spent on sample preparation and counting was also evaluated. MF was more sensitive than MM at ≤ 50 EPG level but not above this while MM had a significantly higher egg recovery rate than MF for ≥ 50 EPG levels (MM = 89.7 %, MF = 68.2 %; P < 0.0001). Operator factors did not have a significant effect (P = 0.358-0.998) on egg counts across methods and EPG levels. The CV between replicates of the MM and MF methods for ≥ 50 EPG was 43.4 and 36.5 %, respectively. The inter-observer CV of the MM and MF methods for ≥ 50 EPG levels was 63.8 and 44.3 % respectively. When the naturally infected free-range layers which were individual caged for excreta sampling, the proportion of samples positive for MM and MF were 91.7 and 96.5 %, respectively (P = 0.023). MM resulted in significantly (P = 0.029) higher excreta egg counts (604) than MF (460) with the difference between methods greatest at higher EPG levels. Fresh floor excreta (pooled or individual) and individual caged chicken excreta did not have significant effect on egg counts (P = 0.274). The total time taken for sample preparation and egg counting was significantly lower using the MM method (4.3-5.7 min) than the MF method (16.9-23.8 min) (P < 0.0001). In conclusion, MM was more accurate than MF, particularly at higher EPG levels, but slightly less precise and sensitive, particularly at low EPG levels, while taking less than 25 % of the laboratory time per sample. Our observations indicate that the MM method is more appropriate for rapid diagnosis of chicken nematodes in the field. Pooled fresh floor excreta samples would be sufficient to indicate infection level in free range farms.