Aeromonas bacteria can cause an infection characterized by septicemia and is one of the most common pathogens in tropical fish. This disease is responsible for high morbidity and mortality rates, causing considerable losses in aquaculture. Thus, the understanding of its pathophysiology is crucial to develop control strategies of this bacterial infection in farmed fish. This study aimed to characterize early pathological aspects of acute sepsis in pacu (Piaractus mesopotamicus) experimentally infected with Aeromonas hydrophila. A total of 160 juvenile pacus were inoculated intraperitoneally with A. hydrophila (1.78 x 109 CFU/mL) and at 0 (control), 1, 3, 6, and 9 hours post-inoculation (hpi), animals were anesthetized and samples were collected for microbiological, light microscopy and transmission electron microscopy (TEM) analyzes. The results showed the occurrence of hemodynamic alterations, such as hemorrhage and congestion, which were observed mainly after 6 and 9 hpi. It was possible to re-isolate Aeromonas at all sampling times except in control group. However, just after 9 hpi it was possible to find the bacteria in all fish and tissues. Light microscopy analyses revealed a degenerative process, necrosis and vascular damage mainly at 6 and 9 hpi. According to the ultrastructural examination, areas of cellular death were identified in all examined tissues, especially at 6 and 9 hpi. However, the most severe, related to necrosis, were observed after 6 and 9 hpi. The findings suggested that this bacterium spreads in the first hpi through the fish organs, mainly affecting spleen, liver and kidney, causing irreversible lesions at the molecular level.
Os objetivos deste trabalho foram os de avaliar o percentual de sobrevivência, a microbiota instestinal, a integridade da mucosa, e a qualidade da carcaça de juvenis de Tilápias-do-Nilo, Oreochromis niloticus, após 80 dias de alimentação com dieta contendo aditivo probiótico (Bacillus cereus 4,0x10(8) UFCg-1 e Bacillus subtilis 4,0x10(8) UFCg-1), na proporção de 4g/kg de ração peletizada. O delineamento experimental foi inteiramente casualizado com dois tratamentos, sendo um grupo controle e outro alimentado com dieta adicionada de probiótico. Foram realizados os cálculos do percentual de sobrevivência relativa, análise da microbiota intestinal por cultura microbiológica, análise histomorfométrica da mucosa intestinal e análise químico-bromatológica da carcaça dos peixes. Os resultados demonstraram que as tilápias do grupo tratado apresentaram percentual de sobrevivência relativa maior (P<0,05) que o do grupo controle e colonização intestinal por B. cereus e B. subtilis com maior (P<0,05) número de unidades formadoras de colônia em relação ao grupo controle. A análise histomorfométrica demonstrou que o grupo alimentado com aditivo probiótico apresentou vilosidades mais altas e mais largas, além de maior número de células caliciformes que o observado no grupo controle (P<0,05). Em relação à qualidade de carcaça os resultados demonstraram que houve interferência positiva (P<0,05) do probiótico no grupo tratado em relação ao controle quanto aos teores de proteína e extrato etéreo. Estes resultados permitem inferir que a suplementação com probiótico, como testado neste estudo, induziu a colonização intestinal por bactérias benéficas e promoveu maior percentual de sobrevivência relativa, diminuiu a descamação da mucosa, e favoreceu o aumento do número de células caliciformes.
The aim of this study was to evaluate the effect of probiotic Bacillus amyloliquefaciens on the growth performance, blood profile and intestinal morphometry in Nile tilapia (Oreochromis niloticus) reared in cages. 936 Nile tilapias were distributed in 12 cages (1.5 m3). Fish were fed for 90 days on basal diets containing 0 (control); 1×106 CFU g-1; 5×106CFU g-1; and 1×107CFU g-1 of the probiotic. The results showed no significant difference on performance and proximal composition of fish. Blood glucose and hemoglobin were lower in 1×107 CFU g-1 group, suggesting improves of the homeostatic state of the fish. Other hematimetric indices did not differ between groups. It was observed significant increase of villi height and in the number of goblet cells of the intestine in fish supplemented 5×106 CFU g-1 and 1×107 CFU g-1 of food suggesting that fish improved the digestion and absorption of nutrients. However, more studies are needed to determine the efficacy of this probiotic in field conditions in Nile tilapia.
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.