Fig 1.
Experimental design of Ascaris suum single- and reinfection.
Fig 2.
Number of larvae recovered from host organs.
(A) Liver on the 4th day post-infection; (B) lung on the 8th day post-infection; (C) BAL on the 8th day post-infection; and (D) gut on the 12th day post-infection. Filled circles–single infection (SI) group; Open circles–reinfection (RE) group. Mann-Whitney test was used to assess differences between groups and are depicted in the graphs by the p values.
Fig 3.
Histopathological visualization of lesions caused by larval migration in the liver on the 4th day post-infection and area of the lesions caused by larval migration.
(A and B) Non-infected mouse; (C and D) Single-infected mouse with the presence of necrosis (arrowheads) and mild inflammatory infiltrate (arrows); (E and F) Reinfected mouse with presence of intense inflammatory infiltrate (arrows) and necrosis (*). Lower magnification Bar scale = 50 μm. Higher magnification Bar scale = 200 μm. (G) Area of lesion caused by larval migration in liver on the 4th day post-infection. Mann-Whitney test was used to evaluate differences between groups.
Fig 4.
Histopathological visualization of the lesion caused by larval migration in the lungs on the 8th day post-infection and area of the lesion caused by larval migration.
(A and B) Non-infected mouse; (C and D) Single-infected mouse with slight thickening of the septum at the expense of inflammatory infiltrates, hyperaemia and haemorrhage (*); (E and F) Reinfected mouse with inflammatory infiltrate, intense hyperaemia and haemorrhage, causing extensive thickening of the septum (*). Lower magnification Bar scale = 50 μm. Higher magnification Bar scale = 100 μm. (G) Area of the lesion caused by larval migration and inflammation in the lung on the 8th day post-infection; Mann-Whitney test was used to assess differences between the groups. (H-I) Optical density representing the MPO and EPO activity in the lung at 8 days post-infection. (H) EPO production in the lung on the 8th day post-infection. (I) MPO production in the lung on the 8th day post-infection. Kruskal-Wallis test followed by Dunn´s multiple comparisons test was used to evaluate differences between groups. The p values in the graphs represent the significant differences.
Fig 5.
Assessment of lung mechanics after single or multiple Ascaris infection in mice.
Forced spirometry was performed to investigate the injury by modifications in lung functions. The parameters assessed were Functional Vital Capacity (A), Inspiratory Capacity (B), Dynamic Compliance Forced (C), Chord Compliance (D), Expiratory Volume at 100 msec (E) and Lung Resistance (F). Kruskal-Wallis test followed by Dunn´s multiple comparisons test was used to evaluate differences among groups. Results are shown as the mean ± SEM. * p < 0.05; ** p < 0.01.
Fig 6.
Levels of haemoglobin, total protein, mononuclear and granulocyte cell counts in the BAL on the 8th day post-infection.
(A) Haemoglobin levels in BAL on the 8th day post-infection. (B) Larvae from A. suum surrounded by leukocytes in the BAL 8 days post-infection. (C) Total protein levels in the BAL on the 8th day post-infection. (D) Total leukocytes counts in the BAL on the 8th day post-infection. (E) Macrophage counts in the BAL. (F) Lymphocyte cell counts in the BAL. (G) Neutrophil cell counts in the BAL. (H) Eosinophil cell counts in the BAL. Kruskal-Wallis test followed by Dunn´s multiple comparisons test was used to evaluate differences among the groups. Results are shown as the mean ± SEM and were represented * and # for was used where * p< 0.05; ** p< 0.01 and *** p< 0.001 for the differences among all groups in the respective time; and # p< 0.05 and ## p<0.01 for differences to the control group at the same time.
Fig 7.
Mononuclear and granulocyte cell counting in the blood at different time points after A. suum experimental infection.
(A) Lymphocyte cell counts. (B) Monocyte cell count. (C) Neutrophil cell counts. (D) Eosinophil cell counts. Filled circles–non-infected group; Open circles–single-infected group; and divided circles–reinfected group. Two-way ANOVA test followed by multiple comparison test were used to compare the variances between the groups. Results are shown as the mean ± SEM and were represented ‘*’ and ‘#’. * p< 0.05; ** p< 0.01 *** p< 0.001 and **** p< 0.0001 represent the differences between all groups in the respective time; and # p<0.05 and ## p<0.01 represent the differences to the non-infected group.
Fig 8.
Systemic cytokine production profile from the first day to 12 days post-infection.
(A) IL-4, (B) IL-5, (C) IL-10, (D) IL-6, (E) IL-17A, (F) IFN-γ, (G) TNF-α and IL-2 (H). Two-way ANOVA test followed by multiple comparisons test were used to compare the variances between the groups. Results are shown as the mean ± SEM and were represented by ‘*’ and ‘#’. * p< 0.05, ** p< 0.01, *** p< 0.001; and **** p< 0.0001 are indicated to represent the differences compared with single-infected group in the respective time; and # p< 0.05, ## p< 0.01, ### p <0.001, and #### p< 0.0001 represent the differences compared with non-infected group.