Promastigote Leishmania-organisms were diagnostically cultivated in vitro from popliteal lymph node aspirates obtained from 32 of in total 36 dogs returning from endemic areas. Isoenzyme analysis (glucosephosphate-isomerase (GPI), phosphoglucomutase (PGM) and glutamate-oxaloacetate-transaminase (GOT) resulted in the identification of Leishmania infantum (syn. Leishmania (L.) infantum) for all 18 isolates investigated. Parasites were still able to be cultivated in vitro in 79% of 28 biopsies (from 15 dogs) even following chemotherapy by Glucantime, independent of the time of sampling and the course of disease after treatment. Dogs with a progressive form of disease (despite chemotherapy) showed only a minor or no reduction (between 0 and 4.8%) of the relative antibody concentration (determined by ELISA), whereas regressive forms of disease (without recurrences observed in the period of 10 to 37 months after therapy) demonstrated a marked reduction of the relative antibody concentration (between 6.7 and 16.2%) within the first 5 to 8 months; thereafter the decrease diminished and changed to a persistent low relative antibody concentration.