Axiocam cc1

Faecal samples were analysed in the Laboratory of Parasitology of the National Institute of Tropical Medicine (INMeT) in Puerto Iguazú, Misiones, Argentina. We processed samples using a semi-quantitative flotation method with a saturated sugar solution^{55 (link)}. We weighted 3 g of faecal sample, then homogenised it, and after centrifugation we mounted it on a slide. We used a Carl Zeiss Primo Star Microscope to identify parasite structures and took pictures with Carl Zeiss AxioCam Cc1 using the 40x magnifier. Eggs and larvae were counted and identified on the basis of colour, shape, content and size. Although most samples reached 3 g after being processed, some samples did not reach this threshold. However, since some parasites were detectable even in < 3 g samples, instead of discarding them, we decided to consider faecal sample’s weight as a control factor in our analyses.

HeLa cells was seeded in 12-well flat-bottomed plates at a density of 1 × 10⁴ cells per well with 1 mL of CM and incubated for 24 h at 37 °C with 5% CO₂. Cells were incubated with serum-free medium (SS) for 12 h at 37 °C and an atmosphere of 5% CO₂ and incubated with different concentrations of the CDPs. After treatment, the cells were washed with PBS. Cells were fixed with paraformaldehyde (PFA at 4%) for 10 min on ice and collocated on cover glass, placed into a holder with a drop of PBS and glycerol 1:1 and photographed using an inverted phase-contrast microscope (Carl-Zeiss HB0-50, Gottingen, Germany) equipped with an AxioCam/Cc1 digital camera (Carl-Zeiss, Gottingen, Germany). Additionally cell cultures were observed directly using a confocal microscope (Olympus FV1000), images of the HeLa cells were taken using 40× magnification.

Alkaline phosphatase blue in situ hybridization stained embryos were mounted for brightfield microscopy in 80% glycerol, 20% PBST, 1 mM EDTA. Images were obtained using an AxioCam CC1 on an AxioPlan2 microscope with a PLAN-NEOFLUAR 20×/0.5 or 10×/0.3 objective and DIC optics using the AxioVs40 Software (all Zeiss). From image stacks (1 μm step size), minimum intensity projections of z-planes as indicated in the figure legends were generated using ImageJ. Embryos stained by fluorescent in situ hybridization and immunohistochemistry were recorded using a Zeiss LSM 510 or LSM 880. All figures were assembled using Adobe Photoshop CS4 or CS6. When linear adjustment of levels was made, histograms were clipped to the same values for experimental and control images.