AÇÃO DA LUZ ULTRAVIOLETA E DA RIBOFLAVINA NA INATIVAÇÃO DE LEISHMANIA EM SANGUE CANINO CONSERVADO EM BOLSAS PARA TRANSFUSÃO

It is extremely important for the transfusion medicine the safety in blood components transfer procedure, minimizing the occurrence of pathogen transmission. This study investigated the efficiency of ultraviolet light and riboflavin in the inactivation of Leishmania infantum chagasi in canine blood samples collected in plastic bags for transfusion. To detect positive blood bags to be used in the experiment, it was performed conventional PCR of the bags collected from symptomatic animals, with positive aspiration of lymph node and IFA (title 1: 640), from an epidemic region for the disease. After 21 days of storage at 4°C, parasitized canine blood riboflavin was added to a final concentration of 50μM. Thereafter, the bag was placed in the illuminator at a wavelength of 365 nm UV light for 30 to 45 minutes and maintained over a homogenizer blood bag. To prove the inactivation, they were used 28 hamsters (Mesocricetus auratus), adult, male. Seven animals were inoculated with the blood without treatment (group leishmaniasis: GL); seven, with blood after the addition of riboflavin (riboflavin group: GRB); seven, with blood after treatment with riboflavin associated with ultraviolet light for 30 minutes (treated group 1: GT30) and seven, with blood after treatment with riboflavin associated with ultraviolet light for 45 minutes (treated group 2: GT45). The animals were kept for 120 days in boxes for hamsters in Infectorio of Zoonoses and Public Healthy, FMVZ, Unesp, Botucatu, and followed clinically. The blood of hamsters was collected by intracardiac puncture for realization of serological and molecular tests. Spleen, liver and bone marrow were extracted after the autopsy, were macerated, and the bone marrow obtained after maceration of the sternum, and DNA extraction was performed to make conventional PCR and qPCR. The parasitized blood has not lost the ability to cause infection after the storage period (21 days). The hamsters inoculated with blood treated with riboflavin and UV light for 30 and 45 minutes were PCR positive, while hamsters not showing clinical signs of the disease. In qPCR, it was identified the association of riboflavin with UV light had reduced the number of Leishmania, but did not completely eliminate the parasites, demonstrating the importance of laboratory diagnosis of leishmaniasis tests in the selection of dogs as blood donors.