Improvement of counting helminth eggs with internal standard

The sanitary regulations introduce a requirement for parasitological control of sewage water used for irrigation and fertilisation of crops. The existing methods of estimation of helminth eggs, based on the coproscopic method using sedimentation, desorption, centrifugation and flotation of the material, allow for only 30-75% efficiency. The aim of this paper is to present an improved quantitative estimation method. The proposed method is based on an internal standard. A predetermined number of eggs of Ascaris lumbricoides suis L is coloured with crystalline violet. These marked eggs are added to sample being examined. Counting the marked eggs in the sample and comparing them with their introduced number allows to estimate the efficiency of the procedure. This results in an increase in the efficiency of the method to 100%. It is possible to calculate the real number of helminth eggs in the sample on the basis of the estimated efficiency of the procedure. The sensitivity of the proposed method is at the level of about 0.4 ppm. The necessary number of replications in the method is about 10 times lower than that in the existing methods. The number of natural (not coloured) eggs does not influence the efficiency of the method with an internal standard.