Characterisation of a new, highly effective method for detecting nematode eggs (Ascaris spp., Toxocara spp., Trichuris spp.) in sewage sludge containing flocculants

Abstract

Because traditional methods used for sewage sludge parasitological examinations have low sensitivity, a new, highly effective method (own method - OM) was devised. The principle of this method is to eliminate the flocculent effect on the structure of sewage sludge by mechanically damaging floccules in the presence of surfactants and to increase the effectiveness of egg isolation processes in large volumes of liquids. The objective of this study was to estimate the effectiveness of the OM in detecting nematode eggs in sewage sludge samples containing flocculants. In the first stage, the effectiveness of the OM was compared to 4 other methods routinely used in parasitological examinations of dehydrated sewage sludge. Next, method standardisation was performed using sewage sludge samples supplemented with eggs from 3 parasite species (Ascaris suum, Toxocara canis and Trichuris vulpis). The study demonstrated that OM efficiency was 6-65 times greater than other methods, depending on the method and type of detected eggs. Limit of detection (LOD) calculations for the OM were performed on samples supplemented with a known number of parasite eggs resulting in 10, 5 and 3 eggs/50 g of sample for A. suum, T. vulpis and T. canis eggs, respectively. The limits of quantification (LOQ) of the OM were established as 200 eggs/50 g of sample for A. suum and T. vulpis eggs and 50 eggs/50 g of sample for T. canis eggs. The rectilinear regression functions, which determined the relationship between the number of eggs detected in OM measurements and the number of eggs contained in the samples, were characterised by high and statistically significant coefficients of determination (r²). The slopes of the trend lines were 0.3188, 0.3821 and 0.3276, and the intercepts were -11.223, -9.0261 and -23.15 for A. suum, T. canis and T. vulpis eggs, respectively. Method sensitivity, calculated as the slope coefficient of the regression function and expressed as a percentage, ranged from 32% to 38% depending on egg type. The study confirmed that the OM may be applied to quantify parasite eggs in dehydrated sewage sludge containing polyelectrolytes.

Keywords: Ascaris spp.; Eggs; Parasitological method standardisation; Sewage sludge; Toxocara spp.; Trichuris spp..