Technical variability and required sample size of helminth egg isolation procedures

Abstract

Measurements of parasite load are often very variable. This implies that little confidence can be attached to single measurements of parasite numbers and egg concentrations, and that many measurements are required for the detection of differences between groups of hosts or parasites. For studies that aim to detect these differences, it is important to increase the precision (closeness of repeated measures to each other) of parasite numbers, because it determines the number of samples that is needed to find significant differences among groups. In this study, sample sizes required to detect group differences were estimated using nematode egg counts of faecal samples of dairy cattle. They were found to be much lower for a centrifugation technique than for the widely used McMaster technique in replicate samples, in spite of a generally similar mean FEC. For example, the sample size required to detect FEC differences between groups of 10, 50, and 250 eggs per gram (EPG) were 46, 25, and 27 for the McMaster technique and 8, 5, and 12 for the SSF method, respectively. Interestingly, sample sizes required for faeces with a relatively high egg concentration (∼1000 EPG) were also considerably lower than for the McMaster technique in spite of a higher mean EPG of the latter method. This implies that technical variation can be reduced considerably by simple methods of egg isolation. Given that the range of egg concentration is similar for a number of nematodes of livestock and human helminths, a reduction of technical error will aid studies with many group comparisons such as vaccination strategies against parasites with typically low FECs and studies of the genetics of host resistance. It may also lead to improved guidelines for measures related to public health.