Caracterização morfológica do intestino do robalo peva (Centropomus parallelus) e avaliação do efeito do Bacillus subtilis sobre mastócitos com grânulos eosinofílicos (MCs/EGC) e células caliciformes intestinais

• Ano de defesa: 2016
• Autor(a) principal: Silva, Wémeson Ferreira da [UNIFESP]
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo (UNIFESP)
• Palavras-chave em Português: Morphology; Immunohistochemistry; Intestinal mucosa; Mast cells; Bacillus subtilis; Sea bass; Morfologia; Imunohistoquímica; Mucosa intestinal; Mastócitos; Robalo
• Link de acesso: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=4016638; http://repositorio.unifesp.br/handle/11600/47308

Characterize anatomical and morphologically different sites and layers of the intestine of fat snook (Centropomus parallelus). Quantify mast cells containing eosinophilic granules (MCs/EGC) and goblet cells after study specimens being fed with probiotic Bacillus subtilis. Material and Methods: A total of 50 animals were used. The morphologic, histochemical and ultra-structural study was carried out on 23 individuals, the histochemical and immune-histochemical was performed on 12 animals and 15 specimens was used as controls for the number of MCs/EGC and goblet cells, and for any variation in number of them after the supplementation of probiotic at the doses of 5g and 10g per kg of regular fish treat. Histological sections for light microscopic study were 5?m thick, were stained with HE dye, toluidine blue (semithin sections of 1?m), Nilceu M. Castro trichrome staining method and silver staining. For histochemical analysis, staining methods comprised PAS, pH-2.5 and pH-0.5 Alcian blue (AB), pH-2.5 AB plus PAS. For MCs/EGC morphology study the methods used were ferric hematoxylin eosin, Congo red, pH-6 acridine orange and toluidine blue. The histochemical analysis was carried out on sections treated with PAS, pH-2.5 and pH-0.5 AB, pH-2.5 AB plus PAS. Antibodies anti-tryptase, chimase and serotonine were used for immunohistochemical study. Tissue fragments 0.5mm-sized, fixed in Karnovsky solution were conventionally processed for Transmission Electron Microscope imaging. MCs/EGC and goblet cells were quantified in one-square millimeter areas of serial sections from sites of the intestine of animals from all 3 study groups. Results: Anatomically, the intestine comprised pyloric caeca, proximal and distal intestine, and rectum. Morphologically, the layer comprised the mucosa, submucosa, muscular layer and serosa. The mucosa presented an epithelium containing absorptive cells (enterocytes), endocrine cells, intraepithelial limphocytes, rodlet cells, goblet cells positive for neutral glycoproteins, acidic carboxylated and sulfated glycosaminoglycans, lamina propria consisted of connective tissue, blood vessels, MCs / EGC, leukocytes and leukocyte infiltrate, and a muscular mucosa between the mucosa and the submucosa. Connective tissue, blood vessels and leucocytic infiltrate formed the submucosa. Their granules of MCs/EGC showed an acidic and basic character and were also metachromatic, presence of neutral glycoproteins, carboxylated and sulfated glycosaminoglycans and positive for tryptase, chymase and serotonine were finding. The muscular layer showed an inner circular and an outer longitudinal arrangement and it?s connective tissue contained nerve plexuses. The serosa was formed by a thin layer of connective tissue, containing vessels and the subserosa nerve plexus. Ultrastructurally, the mucosa showed enterocytes showing an euchromatic nucleus within a cytoplasm containing mitochondria and electrondense spherical granules, goblet cells showing euchromatic nucleus and cytoplasm plentiful of granules of varied size and electrondensity, four types of enteroendocrine cells, intraepithelial phagocytes, rodlet cells showing euchromatic nucleus and rod-shaped cytoplasmic granules of varying electron density and MCs/EGC with euchromatic nucleus and spherical cytoplasmic granules. The quantification of MCs/EGC showed statistically significant difference only for animals in the group 5-grams, which had an increased mean number of these cells when compared with that showed by the control group. Conclusions: The intestine of the fat snook has pyloric caeca, proximal and distal intestines and rectum. Its mucosa contains enterocytes, goblet cells and enteroendocrine cells, intraepithelial lymphocytes, rodlet cells, phagocytes and MCs/EGC. The granules of MCs/EGC had basic and acidic content as well as tryptase, chymase and serotonine. Ultra-structurally, four different types of endocrine cells were identified and the supplementation of probiotic increased the number of MCs/EGC in the intestinal mucosa and submucosa.