In our study, methods for the biomarkers of detoxification and oxidative stress (glutathione GSH, glutathioneStransferase GST, glutathione peroxidase GPx, glutathione reductase GR, lipid peroxidation LP) were introduced and optimized for enchytraeids Enchytraeus albidus and Enchytraeus crypticus. Physiological levels of parameters were characterized and modulations caused by pH, carbendazim, and DMNQ were investigated. Aquatic 96 h test was adopted as suitable exposure assay for testing effects of chemicals on these biomarkers. Numbers of 10 E. albidus and 20 E. crypticus, pH adjusted to 6 to 7, and minimum of three replicates are recommendations derived by experiments in our study. GSH, GR, and GST seemed to be inducible by stress like pH or exposure to toxicants, although stronger effects would be expected from literature data. Our pilot study showed a new direction in enchytraeid ecotoxicology which now needs further research.