Viability assessment of Ascaris suum eggs stained with fluorescent dyes using digital colorimetric analysis.

The aim of the study was to develop a method for the colorimetric evaluation of nematode eggs using appropriate instruments. The materials for the study were live and dead (inactivated) eggs of the Ascaris suum. Viability of the eggs was assessed using four different kits for fluorescent staining (for each technique, a series of photos were taken). Images of stained eggs were analysed using graphic software with RGB (red-green-blue) function. The viability of the eggs was assessed according to the relative positions of the distributions of colour intensities of live or dead eggs - distributions area's overlap index (DAOI), and distributions area's separation index (DASI) were calculated. Computer analysis of the intensity of green colour was not satisfactory. However, analysis of images in the spectrum of red colour proved useful for the effective differentiation between live or dead eggs. The best parameters were observed using the Annexin V FITC Apoptosis Detection Kit (DASI = 41 and 67). The investigation confirmed the usefulness of fluorescent dyes used in conjunction with digital analysis for the assessment of the viability of A. suum eggs. The use of computer software allowed a better objectivity of the assessment, especially in the case of doubtful staining.

First case of Trichinella nativa infection in wild boar in Central Europe-molecular characterization of the parasite.

The examination of wild boars gained in Poland shows for the first time occurrence of Trichinella nativa, freeze-resistant species of Trichinella in this host from the central Europe region. This finding is not only one of several cases of T. nativa invasion in wild boars all over the world but also one of the very few cases of T. nativa detected so far beyond the known boundary of occurrence of this species. The molecular characterization of discovered larvae based on analysis of partial genes: 5s rDNA-ISR and CO1 confirm the findings. Moreover, the analyzed DNA sequences of both genes present new haplotypes of T. nativa in comparison to that described previously.