Synthetic gene as target to assess the sensitivity of PCR to detect Trichinella spp. larvae in meat from a non-endemic region.

Trichinellosis is a zoonotic disease exotic in Brazil but commonly found worldwide including South American countries like Argentina. International trading of swine meat needs an official Trichinella-free diagnosis commonly carried out by pepsin-HCl digestion of diaphragm tissue fragments followed by microscopic examination for the presence or absence of Trichinella larvae. The easiness of this diagnostic method allows it to be performed at slaughtering plants but, in contrast, it lacks sensitivity and does not allow species differentiation, which is fundamental for determining geographical and species distribution of different genotypes. In our study, we aimed to evaluate a highly sensitive diagnostic method based on the polymerase chain reaction (PCR) that would allow us to detect and classify different species of Trichinella. Thus, we designed a synthetic gene and selected five sets of primers targeting specific regions of the Trichinella genome. The synthetic gene was cloned into a plasmid and then used to optimize PCR conditions. Using our PCR, we were able to detect 0.001 pg of the synthetic gene, which corresponded to 0.01 larvae. Then, we collected 175 samples of Suidae (domestic and wild boars) diaphragm fragments that were pooled into groups, digested with pepsin-HCl, and had the DNA extracted for analysis by PCR. The clinical samples evaluated were negative by PCR. Our results indicate that the PCR-based method might be a useful diagnostic method complementary to the pepsin-HCl digestion method currently in use, mostly in non-endemic areas.

ß-Glucan improves wound healing in silver catfish (Rhamdia quelen).

The immune modulating activity of ß-glucan on aquatic species has been a matter of intense investigation. Here, we aimed to investigate the effect of ß-glucan on wound healing of silver catfish, a Neotropical South American scale-free fish. Small sections of skin and muscle (3 mm in diameter) were removed and fish were bathed daily with ß-glucan (0.1% and 0.5%) up to 28 days when cicatrization was complete. A group of fish similarly injured and non-exposed to ß-glucan was used as control. Wound closure and healing was monitored visually and by histopathological analysis. In fish bathed with 0.5% ß-glucan we found reduced blood cortisol levels at day one post-wounding and, by day 7 post wounding, the deposition of granulation tissue was higher compared to non-exposed fish. In addition, from day 7 forward, wound size was significantly lower in fish bathed with 0.5% ß-glucan. Histopathological analysis of the wounded site indicated a thin layer of immature epidermal cells at day one post wounding. A discrete inflammation with mixed inflammatory cell infiltrate was observed on wounded muscle and was lower by day 7 post wounding on fish bathed with 0.5% ß-glucan. By day 14 post wounding, the deposition of collagen fibers and the presence of fibroblast and new muscle fibers were higher in fish exposed to 0.5% ß-glucan, and dermis restoration was complete. Thus, our results indicate that in silver catfish wound healing occurs rapidly and improves greatly by daily bathing with ß-glucan.