No abstract

The direct effect of nitric oxide (NO) on the viability of Toxocara canis larvae was studied. We observed that the nitric oxide donors, SIN-1 and SNOG, exert no cytotoxic effect on the in vitro viability of T. canis larvae. In addition, we developed a model in rats to elucidate the role of NO during T. canis infection. We evaluated different indicators in four experimental groups: morphological parameters, the total number cells and cell types recovered, nitrite and protein concentration, lactate dehydrogenase and alkaline phosphatase enzymatic activity in the bronchoalveolar lavage fluid, lung index and detection of anti-T. canis specific antibodies. We observed significant differences between non-infected and infected groups. The infected animals treated with the inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine were less damaged than infected, non-treated animals. Our results suggest that the in vivo inhibition of the synthesis of NO triggered by iNOS diminishes the deleterious effects of the parasite upon the host, especially the vascular alterations in the lungs. We could show that in vivo production of NO induced by infection with T. canis results in direct host damage. Thus, this induction may constitute an evasion/adaptation mechanism of the parasite.

The effect of four Toxocara canis antigens on nitric oxide (NO) and prostaglandin E2 (PGE2) synthesis was studied in vitro using rat alveolar macrophages. Somatic and excretory/secretory T. canis antigens prepared from adult worms and LII larvae were incubated with rat alveolar macrophages obtained by bronchoalveolar lavage at concentrations of 0.1-50 microg/ml. Both excretory/secretory adult antigen (ESA) and somatic LII antigen (SLII) stimulate the release of nitrites by alveolar macrophages. This effect was specific (inhibited by L-NAME and L-canavanine) and dose-dependent; 30 microg and 10 microg being the most effective concentrations of ESA and SLII, respectively. Western blot and reverse transcriptase-polymerase chain reaction analyses revealed that ESA antigen stimulates the production of NO at transcriptional level. T. canis ESA also stimulated macrophages to produce PGE2 at transcriptional level. The addition of L-canavanine decreased the release of PGE2 significantly, which suggests that NO mediates the production of this prostaglandin. These results indicate that T. canis can stimulate the release of vasodilatory mediators by macrophages of the host.

Infection by Dirofilaria immitis is a rare cause of solitary pulmonary nodule. We describe the second case of transitory pulmonary nodule found by us in the course of a case-finding study, the third one reported in the literature. The detection of cases of transient pulmonary dirofilariotic nodules suggests that human infection is more prevalent than usually thought, and that this kind of manifestation is frequent in human dirofilariosis. Greater awareness is warranted in endemic areas.

Previous studies of ours have demonstrated that a recombinant protein (Fh15) related to fatty acid-binding proteins did not induce significant protection in rabbits challenged 2 or 4 wk postimmunization over nonimmunized controls. In the current study, rabbits were immunized with Fh15 and challenged with Fasciola hepatica metacercariae 12 and 20 wk later. In the current study in which longer lag periods for challenge infection after the second immunization were used, worm burden reductions compared to adjuvant controls were a significant 43% and 76%, respectively. Importantly, rabbits immunized with Fh15 had significant numbers of immature flukes, 66% in the 12-wk period and 84% in the 20-wk lag period as compared to controls. In addition, liver lesions were clearly diminished in the vaccinated rabbits. Enzyme-linked immunosorbent assay absorbance values showed that immunized rabbits developed high antibody levels to Fh15 from 8 wk after the first immunization and did not increase after challenge. These results suggest that a recombinant F. hepatica molecule related to fatty acid-binding proteins induces protective (worm burden reductions), anti-fecundity (immature flukes), and anti-pathology (less liver lesions) effects in rabbits and may serve as a model for the immunoprophylaxis of fascioliasis.