Despite the significant increase in the generation of SARS-CoV-2 contaminated domestic and hospital wastewater, little is known about the ecotoxicological effects of the virus or its structural components in freshwater vertebrates. In this context, this study evaluated the deleterious effects caused by SARS-CoV-2 Spike protein on the health of Danio rerio , zebrafish. We demonstrated, for the first time, that zebrafish injected with fragment 16 to 165 (rSpike), which corresponds to the N-terminal portion of the protein, presented mortalities and adverse effects on liver, kidney, ovary and brain tissues. The conserved genetic homology between zebrafish and humans might be one of the reasons for the intense toxic effects followed inflammatory reaction from the immune system of zebrafish to rSpike which provoked damage to organs in a similar pattern as happen in severe cases of COVID-19 in humans, and, resulted in 78,6% of survival rate in female adults during the first seven days. The application of spike protein in zebrafish was highly toxic that is suitable for future studies to gather valuable information about ecotoxicological impacts, as well as vaccine responses and therapeutic approaches in human medicine. Therefore, besides representing an important tool to assess the harmful effects of SARS-CoV-2 in the aquatic environment, we present the zebrafish as an animal model for translational COVID-19 research.

Camu camu, Myrciaria dubia, is an Amazon plant that presents high levels of vitamin C in its composition. Several studies in animals and humans have demonstrated their efficiency in the prevention and treatment of various diseases. However, there are no reports of its properties in fish. The aim of this study was to evaluate the effect of the oral administration of the extract of this plant in the immune parameters in Nile tilapia, Oreochromis niloticus. 400 Nile tilapia (80 ± 5 g) were randomly distributed into 20 tanks with 1500 L capacity each (20 fish/tank). After a week of adaptation to environmental conditions, it was provided a diet for 5 weeks, using different levels of inclusion of camu camu extract: 0, 50, 100, 250, and 500 mg/kg of feed. Each treatment consisted of four replicates. It was obtained 40.5 mg of vitamin C/g of camu camu pulp powder by high-performance liquid chromatography. At the end of the trial period, fish were inoculated with Aeromonas hydrophila in the swim bladder. Samples were taken after 6; 24 and 48 h of the challenge. Results revealed that fish supplemented with this herb showed significant increase (P < 0.05) in white blood cells counts in blood and exudate, burst respiratory activity, lysozyme activity, serum bactericidal activity, direct agglutination, and melanomacrophage centers count. Red blood cells count, hemoglobin, hematocrit, and biochemical profile of fish supplemented with the herb presented no statistical differences compared to control group (P > 0.05). No histopathological lesions were observed in intestine, kidney, spleen, and gills. It can be concluded that the addition of Myrciaria dubia in tilapia feed improves the immune response and the growth after 5 weeks, especially, at a dose of 500 mg/kg.

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Establishing new experimental animal models to assess the safety and immune response to the antigen used in the development of COVID-19 vaccine is an imperative issue. Based on the advantages of using zebrafish as a model in research, herein we suggest doing this to test the safety of the putative vaccine candidates and to study immune response against the virus. We produced a recombinant N-terminal fraction of the Spike SARS-CoV-2 protein and injected it into adult female zebrafish. The specimens generated humoral immunity and passed the antibodies to the eggs. However, they presented adverse reactions and inflammatory responses similar to severe cases of human COVID-19. The analysis of the structure and function of zebrafish and human Angiotensin-converting enzyme 2, the main human receptor for virus infection, presented remarkable sequence similarities. Moreover, bioinformatic analysis predicted protein-protein interaction of the Spike SARS-CoV-2 fragment and the Toll-like receptor pathway. It might help in the choice of future therapeutic pharmaceutical drugs to be studied. Based on the in vivo and in silico results presented here, we propose the zebrafish as a model for translational research into the safety of the vaccine and the immune response of the vertebrate organism to the SARS-CoV-2 virus.

Environmental changes affect fish homeostasis, turning them more susceptible to diseases. In Brazil, outbreaks of Streptococcus agalactiae infection have been reported in Nile tilapia when they are outside of their thermal comfort zone. This investigation evaluated mortality rate and which were the most infected organs at temperatures that naturally occurred in southern of Brazil. Forty Nile tilapia (Oreochromis niloticus) were infected with S. agalactiae and distributed in four groups (n=10) and each group was exposed to a different temperature: G1: 24°C, G2: 26°C, G3: 28°C, and G4: 32°C. Fish were monitored for 10 days. In this period, fish that presented irreversible clinical signs were sacrificed and samples of brain, liver, and kidney were collected for bacteriological and molecular analysis. Signs compatible with a streptococcal infection were observed in all groups. Highest mortality rates occurred at 24°C and 32°C. The brain was the most affected organ with the highest percentage of isolation of S. agalactiae by both methods of diagnosis. The results suggest that, as in mammals, temperatures that are further away from the comfort zone influence fish homeostasis, increasing susceptibility to bacterial infections. Keywords:Homeostasis. Oreochromis niloticus. Teleost. Thermal stress. ResumoMudanças ambientais afetam a homeostase dos peixes, tornando-os mais suscetíveis a doenças. No Brasil, têm sido relatados surtos de infecção por Streptococcus agalactiae em tilápia do Nilo, principalmente quando se encontram fora da zona de conforto térmico. No presente trabalho, foi avaliada a taxa de mortalidade e determinado quais foram os órgãos mais afetados por essa bactéria em temperaturas que ocorrem naturalmente no Sul do Brasil. Quarenta tilápias-do-nilo (Oreochromis niloticus) foram infectadas por Streptococcus agalactiae e distribuídas em quatro grupos (n = 10), cada um deles submetidos a diferentes temperaturas: G1: 24°C, G2: 26°C, G3: 28°C e G4: 32°C. Os peixes foram monitorados durante 10 dias. Os peixes com sinais clínicos irreversíveis foram sacrificados e coletadas amostras de cérebro, fígado e rins para análise bacteriológica e molecular. Foram observados sinais compatíveis com infecção estreptocócica em todos os grupos. A taxa de mortalidade mais elevada ocorreu nos grupos mantidos nas temperaturas de 24°C e 32°C. O cérebro foi o órgão mais afetado, com a maior percentagem de isolamento de S. agalactiae pelos dois métodos de diagnóstico. Os resultados sugerem que, tal como nos mamíferos, temperaturas que estão mais afastadas da zona de conforto afetam significativamente a homeostase dos peixes, aumentando a sua susceptibilidade para infecções bacterianas. Palavras-chave:Homeostase. Oreochromis niloticus. Teleósteo. Estresse por temperatura.