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Fluke burdens, faecal egg output and hepatic damage were assessed in goats immunised with recombinant cathepsin L1 (rCL1) plus Quil A (n=7) or Quil A alone (control; n=7) and challenged with Fasciola hepatica. There were no significant differences in fluke burdens (56+/-26 vs. 92+/-53), average fluke length (20.9+/-3.0 vs. 21.0+/-3.4mm) or faecal egg output (525+/-533 vs. 758+/-677.8 eggs per gram at 17 weeks post-infection) between vaccinated and infected control groups, respectively, but high individual variability was evident. Morphometric and histopathological studies showed reduced hepatic damage in the vaccinated group compared to the infected control, but high individual variability was also recorded. Further vaccine trials in goats should be carried out using a larger number of animals to evaluate rCL1 as a vaccine for fasciolosis in goats., (Copyright 2009 Elsevier Ltd. All rights reserved.)

Protection against Fasciola hepatica in goats immunized with a synthetic recombinant antigen from Schistosoma mansoni fatty acid-binding protein 14 (rSm14) was investigated by assessing worm burdens, serum levels of hepatic enzymes, faecal egg count and hepatic damage, which was evaluated using gross and microscopic morphometric observation. The nature of the local immune response was assessed by examining the distribution of CD2+, CD4+, CD8+ and γ´+ T lymphocytes along with IgG+, IL-4+ and IFN-γ+ cells in the liver and hepatic lymph nodes (HLN). The goats used consisted of group 1 (unimmunized and uninfected), group 2 [infected control - immunized with Quillaia A (Quil A)] and group 3 (immunized with rSm14 in Quil A and infected), each containing seven animals. Immunization with rSm14 in Quil A adjuvant induced a reduction in gross hepatic lesions of 56.6% (p < 0.001) and reduced hepatic and HLN infiltration of CD2+, CD4+, CD8+ and γ´+ T lymphocytes as well as IL-4+ and IFN-γ+ cells (p < 0.05). This is the first report of caprine immunization against F. hepatica using a complete rSm14 molecule derived from S. mansoni. Immunization reduced hepatic damage and local inflammatory infiltration into the liver and HLN. However, considering that Quil A is not the preferential/first choice adjuvant for Sm14 immunization, further studies will be undertaken using the monophosphoryl lipid A-based family of adjuvants during clinical trials to facilitate anti-Fasciolavaccine development.

Worm burden, hepatic damage and local cellular and humoral immune responses were assessed in goats immunized with glutathione-S-transferase and challenged with Fasciola hepatica. Infected but unimmunized and uninfected control groups were also studied. Hepatic damage was evaluated grossly and microscopically. Local immune response was evaluated by (1) microscopical examination of hepatic lymph nodes (HLNs); (2) analysis of the distribution of CD2(+), CD4(+), CD8(+), T-cell receptor gammadelta(+) lymphocytes and immunoglobulin (Ig) G(+) plasma cells; and (3) investigation of the distribution of cells expressing interleukin (IL)-4 and interferon (IFN)-gamma in the hepatic inflammatory infiltrates and HLNs. Immunized animals did not have significant reduction in fluke number, but there was significant (P<0.05) reduction of fluke size relative to the control groups. The lesions in the two infected groups were similar and consisted of fibrous perihepatitis and white tortuous tracts, mainly involving the left hepatic lobe. Microscopical lesions were similar in both infected groups and were typical of chronic fascioliosis. These included portal fibrosis, inflammatory infiltration with plasma cells, formation of lymphoid follicles, accumulation of haemosiderin-laden macrophages and granulomatous foci. Both infected groups had a marked local immune response characterized by infiltration of CD2(+), CD4(+) and CD8(+) T lymphocytes, and IgG(+) plasma cells in hepatic lesions and in HLNs. There was no expression of IL-4 or INF-gamma by cells in the hepatic inflammatory infiltrate, but expression of INF-gamma in HLNs was much lower than that of IL-4, suggesting an immune response dominated by T helper 2 cells., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Protection against Fasciola hepatica in goats immunized with Peroxiredoxin (Prx) was assessed. The experimental trial consisted of three groups of seven animals; group 1 were unimmunized and uninfected, group 2 were immunized with adjuvant only and group 3 were immunized with recombinant Prx in adjuvant (immunized and infected). Immunization with Prx in Quil A adjuvant, group 3, induced a reduction in fluke burden of 33.04% when compared to adjuvant control, group 2, although this difference was not significant. The hepatic gross and microscopical morphometric study revealed lower damage in the Prx-immunized compared to group 2 (p<0.05). Furthermore, immunohistochemical studies revealed that the Prx-immunized group exhibited reduced infiltration of CD4(+), CD8(+), IFN-gamma(+) and TCR(+) (p<0.05); and CD2(+) and IL-4(+) (p<0.001) in hepatic lesions. Levels of anti-Prx serum IgG in group 3 showed a significant increase at the 4th week after challenge infection compared with group 2 (p<0.0001). This is the first report of ruminant immunization with recombinant Prx of F. hepatica. The study shows that this vaccine significantly reduces hepatic damage and encourages further studies to improve the vaccine efficacy., ((c) 2010 Elsevier Ltd. All rights reserved.)

The nature of the local immune response was assessed studying the distribution of CD2(+), CD4(+), CD8(+), gammadelta(+) T lymphocytes, IgM(+) B cells, IL-4(+) and IFN-gamma(+) cells in the liver and hepatic lymph nodes (HLN) of goats immunised with a synthetic peptide of the Sm14 antigen from Schistosoma mansoni and challenged with Fasciola hepatica. A morphometric study of HLN was also carried out in order to evaluate the hyperplasia of lymphoid follicles. Despite the decrease in fluke burdens found in the immunised group (45.9%) respect to the infected control group, this difference was not statistically significant due to the high individual variability. In liver, a significant increase of CD2(+), CD4(+), CD8(+), gammadelta(+) T lymphocytes was found in the infected groups respect to the uninfected control and in the infected control respect to the immunised group. HLN showed a significant enlargement due to the hyperplasia of lymphoid follicles and infiltration of CD2(+), CD4(+), CD8(+), gammadelta(+) T lymphocytes in both infected groups respect to the uninfected control, with no significant differences between the infected control and immunised group. IFN-gamma(+) lymphoid cells was absent or very occasional in HLN where the number of IL-4(+) cells was higher than that of IFN-gamma, suggesting a polarized Th2 response in immunised and in infected control group.

Two flocks of broiler chickens aged 15 to 30 days presented respiratory signs such as dyspnea and up to 25% mortality. These were the only two flocks in the farm where a bed of sunflower shells was used instead of the rice-hull bedding used in other flocks. At necropsy, severe ascites, right heart hypertrophy, pulmonary congestion, and extensive multifocal granulomatous pneumonia were recorded. Histopathologic examination revealed chronic multifocal mycotic granulomatous pneumonia. Aspergillus fumigatus was identified by microbiologic study from pulmonary specimens. After disinfecting the floor and changing the bedding, no clinical signs were recorded in the farm. Severe chronic granulomatous pneumonia caused by A. fumigatus in the chickens of the present study may have caused hypoxia, leading to pulmonary hypertension, heart failure, and ascites.

The expression of iNOS by macrophages in 33 dogs suffering from spontaneous leishmaniasis was analysed by immunohistochemistry in skin, liver and lymph nodes. A correlation study between the number of macrophages expressing iNOS and the number of macrophages containing leishmania amastigotes was carried out. Formalin-fixed paraffin-embedded tissue sections from the skin (28 cases), popliteal lymph nodes (8 cases) and liver (3 cases) of dogs of different age, sex and breed suffering from leishmaniasis were included in the study. Dogs were referred as positive for Leishmania spp by serology and the diagnosis was confirmed by demonstration of leishmania amastigotes within macrophages by histopathology. Tissue samples of skin (3 cases), popliteal lymph nodes (5 cases) and liver (3 cases) from dogs seronegative for leishmaniasis with no histopathological changes were included in the study as controls. The immunohistochemical study revealed that macrophages containing a high number of leishmania did not express iNOS. Correlation between the number of macrophages expressing iNOS and the number of macrophages containing leishmania amastigotes was assessed using the Spearman test. High expression of iNOS in macrophages was related with low number of leishmania amastigotes in macrophages in all cases (r=-0.47, p=0.002). These results suggest that iNOS expression by macrophages plays an important role during the control of Leishmania infection in dogs.