No abstract

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.

No abstract

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.

The anthelmintic efficiency of doramectin, fenbendazole, and nitroxynil, used individually or in combination, was determined by the Fecal Egg Count Reduction (FECR) test and cultivation of larvae of anthelminthic-treated sheep grouped as follows: G1 (doramectin), G2 (fenbendazole), G3 (nitroxynil), G4 (doramectin + fenbendazole), G5 (doramectin + nitroxynil), G6 (fenbendazole + nitroxynil), G7 (doramectin + nitroxynil + fenbendazole), G8 (untreated). In addition to individually used doramectin and fenbendazole, the helminths were also resistant to the combination of doramectin + fenbendazole; nitroxynil + fenbendazole; and doramectin + nitroxynil + fenbendazole, with their FECR rates ranging from 62-83%. The helminths showed possible nitroxynil-resistance, but had low resistance when the drug was administered in combination with doramectin. The evaluation of individual helminth species revealed that fenbendazole was fully effective against Cooperia; doramectin (G1), moderately effective against Haemonchus and insufficiently active against Cooperia; nitroxynil, effective against Haemonchus and insufficiently active against Cooperia. It was concluded from the results that herd nematodes are resistant to doramectin, fenbendazole, and nitroxynil, and that the combined use of the drugs not only fails to significantly improve the anthelmintic efficiency against Haemonchus and Cooperia, but is also cost-ineffective.Keywords: Parasitic resistance, Haemonchus, Cooperia, small ruminants, anthelmintic association. ResumoEficiências da doramectina, fenbendazole e nitroxynil, utilizados individualmente ou associadamente, foram determinadas através do Teste de Redução na Contagem de Ovos nas Fezes (RCOF) e cultivo de larvas. Os grupos experimentais foram os seguintes: G1 (ovinos tratados com doramectina), G2 (fenbendazole), G3 (nitroxynil), G4 (doramectina + fenbendazole), G5 (doramectina + nitroxynil), G6 (fenbendazole + nitroxynil), G7 (doramectina + fenbendazole + nitroxynil) e G8, não tratados. Os helmintos foram considerados resistentes a doramectina e ao fenbendazole isoladamente e às associações doramectina + fenbendazole, fenbendazole + nitroxynil, e doramectina + fenbendazole + nitroxynil, com taxas de RCOF variando de 62-83%. Helmintos foram considerados suspeitos de resistência ao nitroxynil e apresentaram baixa resistência, quando esta droga foi associada à doramectina. Dos tratamentos isolados, o fenbendazole demonstrou total eficácia (100%) contra Cooperia. Doramectina (G1) foi moderadamente efetiva contra Haemonchus e insuficientemente ativa contra Cooperia, e o nitroxynil efetivo contra Haemonchus (93,2%) e insuficientemente ativo contra Cooperia (0%). Concluiu-se neste estudo que os nematódeos do rebanho são resistentes à doramectina, fenbendazole e nitroxynil, e que, ainda que associadas, não devem ser utilizadas no rebanho por não melhorarem a eficiência anti-helmíntica nem a efetividade contra Haemonchus e Cooperia e por não apresentarem custo-benefício justificado.