RESUMO
Foot-and-mouth disease (FMD) is considered as contagious in livestock, which is caused by the Picornavridae virus family known as FMD virus (FMDV). In the present study, the VP1 gene from FMDV (O strain) was expressed and purified. In addition, nanoliposomes were utilized as an adjuvant. After formulating the nanoliposomes with DMPC, DMPG, and cholesterol, the recombinant VP1 protein was encapsulated in the nanoliposomes. Further, the intended nanoliposomes in the sizes of 400 and 200 nm, nanoliposome-inactivated FMDV, Freund's adjuvant-inactivated FMDV, and Freund's adjuvant-recombinant VP1 mixtures, and PBS buffer (negative control) were injected to six groups of five guinea pigs. Furthermore, the guinea pig serums were analyzed through using ELISA and serum neutralization tests after four boosting vaccinations. Based on the results, the immunogenicity of the 200 nm-nanoliposomes encapsulating recombinant VP1 protein was more than that of 400 nm-ones so that 200 nm-nanoliposomes could trigger the immune response against FMDV in guinea pigs.
Assuntos
Vírus da Febre Aftosa , Febre Aftosa , Vacinas Virais , Cobaias , Animais , Adjuvante de Freund , Proteínas do Capsídeo/genética , Anticorpos Antivirais , Febre Aftosa/prevenção & controle , Modelos AnimaisRESUMO
Background: Autolysate products from yeast origin are very interesting for food, feed, cosmetic, pharmaceutical, and fermentation industries. The lysis process greatly influences the quality and efficiency of the final autolysates. Objectives: Here, we have compared four lysis methods based on autolysis, plasmolysis (with ethanol 1.5% (v/v) and coconut fatty acids 1% (w/w)) and hydrolysis (with alkaline protease 0.4 % (v/w)) on degrading the baker's yeast Saccharomyces cerevisiae. Materials and Methods: The efficiency of processes was evaluated according to the recovered solid and protein contents, release of intracellular materials, cell viability, microscopy imaging, degree of hydrolysis and electrophoresis studies. Results: Results showed that the increased recovered solids and proteins, as well as a higher degree of hydrolysis (DH) were obtained for the enzymatic hydrolyzed cells using alkaline protease. SDS-PAGE analysis also confirmed the results. Further, functionality of the final products by agglutination test showed that the hydrolyzed cells could effectively bind pathogenic bacteria compared to the other cell lysates. Conclusions: In conclusion, this work provides adequate evidence for efficiency of alkaline protease for producing the nutritional cell lysates from baker's S. cerevisiae used in food, feed, cosmetic, and pharmaceutical applications. Moreover, this was the first report on using coconut fatty acids and alkaline protease in lysis of baker's yeast.