A case of canine ocular onchocercosis in Portugal.

Onchocercosis is a newly recognized disease in dogs that has been reported with higher frequency in Europe and in the United States. We report a case of a 3-year-old male mongrel stray dog from the Algarve region (South Portugal) who had a retrobulbar granuloma containing a filaroid nematode of the genus Onchocerca. A gravid adult female parasite was embedded in a granulomatous inflammation adjacent to the sclera beyond the retina. The parasite was 191 to 267 mum in diameter (mean = 225 mum), surrounded by a cuticule and owing a uterus that was filled with small unsheated microfilariae. The cuticule consisted of two separated layers in longitudinal sections. The external layer had cuticular ridges and the internal layer contained striations. Sequencing of the COI and ND5 mitochondrial genes confirmed the identity of this parasite as Onchocerca lupi. Furthermore, the first sequence of the 12S mitochondrial gene is reported in this study.

Application of fluorescence in situ hybridisation (FISH) to the analysis of yeast population dynamics in winery and laboratory grape must fermentations.

To analyse the yeast population diversity during wine fermentations, specific fluorescein-labelled oligonucleotide probes targeted to the D1/D2 region of the 26S rRNA of different yeast species known to occur frequently in this environment were designed and tested with reference strains. The probes were then used to identify wine must isolates and to follow, in combination with plate counts, the evolution of yeast populations in two winery fermentations of white and red grape musts. In both cases, a high diversity of non-Saccharomyces yeast species was detected, including Candida stellata, Hanseniaspora uvarum, H. guilliermondii, Kluyveromyces marxianus, K. thermotolerans and Torulaspora delbrueckii. Some of these species (e.g., K. marxianus, K. thermotolerans and T. delbrueckii) were present in significant amounts during the tumultuous fermentation stage, despite the predominance of Saccharomyces cerevisiae cells following the inoculation of the wine musts with a starter strain. To further clarify the yeast population dynamics at the late phase of the fermentations, and because winery conditions do not allow a reliable control of experimental variables, strains isolated from the industrial musts were used to conduct two laboratory microvinifications in synthetic grape juice, using different ratios of S. cerevisiae/non-Saccharomyces in the inocula. Under these conditions, the results were similar to those obtained in the winery, showing a yeast profile with mixed species throughout the first fermentation stage, i.e. until about 40-50% of the total sugar was consumed. Non-Saccharomyces yeasts were outgrown by S. cerevisiae only after ethanol reached concentrations around 4-5% (v/v), which argues in favour of a potential important role of non-Saccharomyces in the final organoleptic characteristics of the wine.