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The serum pepsinogen level is widely accepted as a useful parameter for monitoring gastrointestinal nematode infections in first-season grazing calves. However, several aspects, with possible implications for its practicality and the way to use the pepsinogen test, have received little attention to date. The objectives of this study were to evaluate (1) the reproducibility of a pepsinogen assay; (2) the required sample size for a reliable diagnosis and (3) the compliance to and effect of advice based on pepsinogen levels determined in year one on the nematode infection levels in the next generation of calves. Despite a high repeatability of the pepsinogen assay within the reference lab, the reproducibility between different labs was poor. There was more variation in pepsinogen levels between herds than between animals within a herd, suggesting that it is most useful to make a herd level diagnosis. Sample size calculation indicated that sampling seven animals sufficed to obtain a reliable indication of calf groups of up to 40 animals. Eighty-two herds were followed-up over two consecutive years and 39 (48%) had followed the advice that was formulated based on pepsinogen levels. Thirty-nine percent of the herds were advised to reduce the intensity of chemoprophylaxis. Samples from animals of those herds showed a higher pepsinogen level in the second year, but none of them had a mean pepsinogen level that is considered excessive. It is concluded that pepsinogen determination at housing from a limited number of animals can contribute to the design of the anthelmintic control strategy in the next year and a more targeted use of anthelmintics. However, more efforts are needed to harmonize the assay between veterinary diagnostic labs.