Detection of Leishmania infantum DNA in the non-parasitized lung of dogs with visceral leishmaniasis.

BACKGROUND: Despite the very low or absent parasitism in the lungs, the interstitial pneumonitis is a common lesion found in humans and dogs with visceral leishmaniasis. The lung is a neglected organ in the study of dogs and humans with visceral leishmaniasis, but interstitial pneumonitis represents an important lesion characterized by thickening of the alveolar septum due to fibrosis and inflammatory exudate, and its pathogenesis is still uncertain. In this study, the polymerase chain reaction (PCR) was used to detect Leishmania infantum in paraffin-embedded lung biopsies from naturally infected dogs from an endemic area in Minas Gerais State, Brazil; PCR was compared to histological and immunohistochemical techniques for detecting Leishmania. RESULTS: Eighteen dogs in which leishmaniasis had been diagnosed by serological tests - indirect immunofluorescence assay (IFAT), enzyme-linked immunosorbent assay (ELISA) and complement fixation tests (CFT) - were classified as asymptomatic, oligosymptomatic or symptomatic. Nine of the 18 dogs studied had a positive PCR (50%) but parasites were not detected by histopathological and immunocytochemistry methods. CONCLUSIONS: These data indicate that PCR on DNA extracted from paraffin-embedded tissue is a valuable method for detecting Leishmania infantum parasites in lungs of naturally infected dogs, despite the apparent absence of parasites from standard HE (hematoxylin and eosin) stained slides and of labeled parasites from immunocytochemical preparations.

Leishmania infantum is present in vaginal secretions of naturally infected bitches at lower levels in oestrogenized bitches than in non-oestrogenized bitches.

Naturally Leishmania infantum infected bitches were divided into oestrogenized (n = 11) and non-oestrogenized (n = 6) groups. Vaginal secretions were collected for polymerase chain reaction (PCR), and vulval, vaginal and uterine tissues were collected for the immunohistochemical (IHQ) identification of L. infantum. Parasite DNA was identified in vaginal secretions of non-oestrogenized (41.8%) and oestrogenized (18.2%) bitches (P<0.05; Fisher's Exact test). IHQ was positive in vulvar dermis (23.5%) and vaginal mucosa (17.7%) but negative in endometrium of all bitches. Poor association between positive vaginal secretion PCR and tissue IHQ (Kappa index) were observed. The results showed that genital secretions are a potential source for dog contamination.

Identification of virulence factors in Leishmania infantum strains by a proteomic approach.

Knowledge of Leishmania virulence is essential for understanding how the contact between the pathogen and host cells can lead to pathogenesis. Virulence in two L. infantum strains was characterized using macrophages and hamsters. Next, we used difference gel electrophoresis (DIGE) and mass spectrometry to identify the differentially expressed proteins. A total of 63 spots were identified corresponding to 36 proteins; 20 were up-regulated, in which 16 had been previously associated with Leishmania virulence. Considering our results and what has been reported before, we suggest the hypothesis that L. infatum virulence could be a result of the increased expression of KMP-11 and metallopeptidase, associated with an improved parasite-host interacting efficiency and degradation of the protective host proteins and peptides, respectively. Other factors are tryparedoxin peroxidase and peroxidoxin, which protect the parasite against the stress response, and 14-3-3 protein-like, which can prolong infected host cell lifetime. Proteins as chaperones and endoribonuclease L-PSP can increase parasite survival. Enolase is able to perform versatile functions in the cell, acting as a chaperone or in the transcription process, or as a plasminogen receptor or in cell migration events. As expected in more invasive cells with high replication rates, energy consumption and protein synthesis are higher, with up-regulation of Rieske iron-sulfur protein precursor, EF-2, S-adenosylhomocysteine, and phosphomannomutase.