title: Real-time PCR assay in Leishmania-infected dogs treated with meglumine antimoniate and allopurinol.
abstract: A real-time PCR assay was exploited for monitoring the Leishmania DNA load in different tissues from 18 naturally-infected dogs before and after treatment with a combination of meglumine antimoniate (100mg/kg/day, subcutaneously) and allopurinol (10mg/kg/day, orally) for 30 days. After the combined therapy, allopurinol was continued at the same dose until the end of the observation period. Whole blood samples, lymph node aspirates, and skin biopsies were collected at the time of diagnosis, 1 month after starting therapy, and every 3 months for 2 years. In six dogs parasite load assessments continued every 6 months for a further 3 years. At each assessment, the dogs were examined for signs of disease and a clinical score was recorded. At diagnosis, the highest Leishmania DNA load was detected in lymph node aspirates. From 1-6 months post-therapy a general improvement in clinical conditions was recorded in all dogs, which correlated with a decrease in the parasite DNA load in all tested tissues, even though it was less pronounced in lymph node aspirates. In the period from 9-24 months post-therapy, a re-increase in parasite load was observed in the tissues of some dogs, concomitant with a disease relapse. The results show that the combined therapy with meglumine antimoniate and allopurinol promoted a clinical improvement which was accompanied by a reduction in the parasitic load in the blood, skin and lymph nodes but, even after long period of allopurinol administration alone, Leishmania may persist in dog tissues.
authors.affiliation: Dipartimento di Scienze Cliniche Veterinarie, Università di Napoli Federico II, Napoli, Italy. laumanna@unina.it
authors.forename: Laura
authors.initials: L
authors.lastname: Manna
authors.forename: Stefano
authors.initials: S
authors.lastname: Reale
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Vitale
authors.forename: Esther
authors.initials: E
authors.lastname: Picillo
authors.forename: Luigi Michele
authors.initials: LM
authors.lastname: Pavone
authors.forename: Angelo Elio
authors.initials: AE
authors.lastname: Gravino
mesh_terms: D000493:Allopurinol / Q000008:administration & dosage / Q000627:therapeutic use*; D000818:Animals; D000981:Antiprotozoal Agents / Q000627:therapeutic use*; D016054:DNA, Protozoan / Q000302:isolation & purification; D004283:Dog Diseases / Q000188:drug therapy*; D004285:Dogs; D004359:Drug Therapy, Combination; D007891:Leishmania / Q000302:isolation & purification; D007896:Leishmaniasis / Q000188:drug therapy / Q000662:veterinary*; D008198:Lymph Nodes / Q000469:parasitology; D008536:Meglumine / Q000008:administration & dosage / Q000627:therapeutic use*; D000077485:Meglumine Antimoniate; D009942:Organometallic Compounds / Q000008:administration & dosage / Q000627:therapeutic use*; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012867:Skin / Q000469:parasitology
chemical_list: D000981:Antiprotozoal Agents; D016054:DNA, Protozoan; D009942:Organometallic Compounds; D000493:Allopurinol; D008536:Meglumine; D000077485:Meglumine Antimoniate