Expanding the known geographic range of Leishmania (Mundinia) enriettii: clinical and molecular characterization in naturally infected guinea pigs from southern Brazil

ABSTRACT: Three domestic guinea pigs (Cavia porcellus) from Ampére, Paraná State, Brazil, presenting crusted nodular lesions on the pinnae, snout, and digits of all limbs were examined at the University Veterinary Hospital of the Federal University of the Southern Border in February 2023. Cytological evaluation by fine-needle aspiration revealed Leishmania amastigotes within macrophages. Two animals were euthanized for further histopathological, immunohistochemical, parasitological, and molecular investigations. Skin samples exhibited a diffuse mixed inflammatory infiltrate, predominantly histiocytic, with macrophages densely parasitized by amastigotes. Immunohistochemistry (IHC) using an anti-mTXNPx monoclonal antibody confirmed the abundant presence of amastigotes in the skin, providing key diagnostic support. Liver sections showed mild lymphocytic infiltrates, with specific IHC labeling in pericentral hepatocytes indicating antigenic presence despite the absence of visible parasites. Kidneys, lungs, and spleen exhibited mild architectural changes with no detectable amastigotes on microscopy. Promastigote forms were successfully isolated in NNN/LIT medium from one liver sample. Molecular analysis using hsp70-targeted PCR and sequencing confirmed the presence of Leishmania DNA in the skin, spleen, and liver, identifying the species as Leishmania (Mundinia) enriettii. The absence of microscopically visible amastigotes in tissues, despite molecular and immunohistochemical evidence, underscores the value of a multimodal diagnostic approach. This study expands the known geographic distribution of L. enriettii in Paraná State and reinforces the guinea pig’s role as the only confirmed reservoir of this parasite in Brazil.