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Title: Avaliação da resposta inflamatória gastroduodenal pela imuno-histoquímica associada a investigação de Helicobacter spp. em cavalos de corrida
Keywords: Gastroduodenite;  Equino;  Helicobacter spp;  Cavalo de corrida;  Imunohistoquímica;  Helicobacter;  Gastroduodenitis;  Racehorse;  Immunohistochemistry
Abstract: Gastroduodenites and gastroduodenal ulceration are frequent diseases of Thoroughbred racehorses. They bring discomfort to the animal and economical losses due to a drop in performance. It is speculated that both the training race and Helicobacter spp. infection can play an important role in the pathogenesis of gastroduodenal disease. In addition, in Brazil, there are no reports of Helicobacter spp. infection of the horse gastric mucosa and there are very few researches about it around the world. The aim of this study was to evaluate by immunohistochemistry the inflammatory gastroduodenal reaction and investigate Helicobacter spp. in the gastroduodenal mucosa of Thoroughbred racehorses. Necropsy was performed in 15 horses and samples of the gastric regions (aglandular fundus, glandular fundus, margo plicatus, antrum, pylorus and duodenum) were obtained. Urease rapid test, direct bacterioscopic exam, histopathology (H&E and Warthin-Starry stains), and Immunohistochemistry (anti- H.pylori, anti-Myeloperoxidase, anti CD3 and anti-CD20 antibodies) were performed. The macroscopic changes found in these animals were mucus, fibrin exudate, blood, edema, erythema, hyperkeratosis and hemorrhage. Histopathology revealed mild to severe diffuse lymphoplasmacytic infiltrate throughout the mucosa sometimes accompanied by areas of polymorphonuclear infiltration, a feature that shows activity. In addition, all animals showed some degree of acanthosis and hyperkeratosis in the glandular stomach. Erosions and ulcers frequencies were 64.3% and 19.6% respectively. Urease activity was detected in five of the nine tested animals. Rods or even round coccoid-shaped organisms were detected in 10 animals. Warthin-Starry stain showed silver impregnated coccus and curved rods in all horses. Immunohistochemistry detected Helicobacter spp. in the glandular and squamous mucosa of 14 horses. The antrum was the most colonized region. Warthin Starry and Immunohistochemistry had statistically similar results. The gastroduodenal region with the highest number of polymorphonuclear cells (MPO +) per field was the margo plicatus. And the duodenum was the gastroduodenal region with the highest number of T (CD3 +) and B (CD20 +) lymphocytes per field. Helicobacter spp. infection was associated with higher numbers of CD3 positive cells infiltrating the gastroduodenal mucosa. Therefore, the horses are susceptible to natural infection with Helicobacter spp.. The inflammatory cell infiltration is closely linked to the kind of histopathologic changes, for example, erosion and deposition of fibrin are associated to an increased infiltration of CD3 and CD20 positive cells, while ulcers, increase the number of MPO-positive cells . Helicobacter spp. colonization is associated with T lymphocytes infiltration. Moreover, several factors may be related to the development of gastroduodenal lesions. Further studies on that matter are necessary in order to establish the relationship between Helicobacter spp., gastroduodenal disorders and inflammatory cells in this pathogenesis
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