ABSTRACT. This work is a sequence of studies on tropical fish of economic importance that evaluated the effects of two different stress of handling on the physiology and haematology of Oreochromis niloticus L. acclimated for 10 days before the essay. The stress consisted in net capture of all fish from each aquarium for 30s emersion. Fish exposed to single stress (SS) the samples were collected in the times 0, 10, 20, 30, 40, 50, 60, 120, 180, 240 and 300min. after stress. In the consecutive stress (CS) the samples were collected in the times 0; 15min. after the first stress; 15min. after the second stress; 15min. after the third stress and 15, 30, 45, 60, 120, 180 e 240min. after the fourth stress totalizing four stimuli every 60min. Fish exposed to SS showed increased cortisol and glucose concentrations at 60min. as well as in the leucocytes number and hematocrit at 50min. after stress. Cortisol did not alter in fish exposed to CS, but glucose increased 15min. after the third stress. On the other hand, CS provoked reduction in the leucocytes number and later hematocrit increasing. Neutrophilia and lymphopenia were related to SS and CS.Key words: Oreochromis niloticus, consecutive stress, cortisol, glucose, haematology. RESUMO. Resposta fisiológica e hematológica de Oreochromis niloticus (Osteichthyes:Cichlidae) exposto ao estresse único e consecutivo de captura. Este trabalho é seqüência de estudos com peixes tropicais de importância econômica avaliando os efeitos de dois tipos de estresse sobre a fisiologia e hematologia de O. niloticus L, aclimatados durante 10 dias antes do experimento. O estresse consistiu na captura de todos os peixes do aquário com rede e emersão por 30 s. Nos animais submetidos ao estímulo único de captura (EU) as amostras foram coletadas nos tempos 0, 10, 20, 30, 40, 50, 60, 120, 180, 240 e 300min. após o estresse. No estímulo consecutivo (EC) as amostras foram coletadas nos tempos 0; 15min. após o primeiro estresse; 15min. após o segundo estresse; 15min. após o terceiro estresse e 15, 30, 45, 60, 120, 180 e 240min. após o quarto estresse totalizando quatro estímulos a cada 60min. Os peixes expostos ao EU apresentaram aumento nas concentrações de cortisol e glicose 60min., bem como no número de leucócitos e hematócrito 50min. após o estresse. Não houve alteração do cortisol nos peixes do EC, mas a glicose aumentou 15min. após o terceiro estresse. Por outro lado, o EC provocou redução no número de leucócitos e aumento tardio do hematócrito. Observou-se neutrofilia e linfopenia após o EU e EC.

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The present study aimed to describe and characterize the cellular components during the evolution of chronic granulomatous inflammation in the teleost fish pacus (P. mesopotamicus) induced by Bacillus Calmette-Guerin (BCG), using S-100, iNOS and cytokeratin antibodies. 50 fish (120±5.0 g) were anesthetized and 45 inoculated with 20 μL (40 mg/mL) (2.0 x 106 CFU/mg) and five inoculated with saline (0,65%) into muscle tissue in the laterodorsal region. To evaluate the inflammatory process, nine fish inoculated with BCG and one control were sampled in five periods: 3rd, 7th, 14th, 21st and 33rd days post-inoculation (DPI). Immunohistochemical examination showed that the marking with anti-S-100 protein and anti-iNOS antibodies was weak, with a diffuse pattern, between the third and seventh DPI. From the 14th to the 33rd day, the marking became stronger and marked the cytoplasm of the macrophages. Positivity for cytokeratin was initially observed in the 14th DPI, and the stronger immunostaining in the 33rd day, period in which the epithelioid cells were more evident and the granuloma was fully formed. Also after the 14th day, a certain degree of cellular organization was observed, due to the arrangement of the macrophages around the inoculated material, with little evidence of edema. The arrangement of the macrophages around the inoculum, the fibroblasts, the lymphocytes and, in most cases, the presence of melanomacrophages formed the granuloma and kept the inoculum isolated in the 33rd DPI. The present study suggested that the granulomatous experimental model using teleost fish P. mesopotamicus presented a similar response to those observed in mammals, confirming its importance for studies of chronic inflammatory reaction.

Summary The study objective was to make histological, histochemical and morphometric evaluations on the splenic Melanomacrophage centers (MMCs) of tilapias, Oreochromis niloticus (Linnaeus, 1758), that were subjected to chronic inflammation stimuli by implantation (IMP) of a glass coverslip in the subcutaneous tissue and through inoculation of the bacillus Calmette‐Guerin (BCG). Randomly distributed in four groups were 150 tilapias: IMP (n = 45); IMP+BCG (n = 45); BCG (n = 45); and control (n = 15). Nine fish per treatment and three control fish were sampled on days 3, 7, 14, 21 and 33. The results demonstrated that increased numbers and areas of these structures were related to the type of stimulus, and that these were greater for the specific response. The principal pigment component identified was hemosiderin. Results suggest that the intensity of the MMC response in O. niloticus depended on the type of inflammatory stimulus used, and that it was greater in fish inoculated with BCG, which induced a granulomatous inflammation when compared to the foreign body inflammatory response induced by the glass coverslips.

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