Mycobacterium bovis BCG expressing the proteins CP40 or CP09720 of Corynebacterium pseudotuberculosis promotes protection in mice after challenge.

In this study, we assessed the survival and immune response of mice vaccinated with recombinant Mycobacterium bovis BCG Pasteur that expressed the CP40 or CP09720 proteins after the mice were challenged with a C. pseudotuberculosis MIC-6 virulent strain. Six groups of mice (n = 10 mice per group) were immunised with a sterile 0.9% saline solution (G1), 106 CFU of M. bovis BCG Pasteur (G2), 106 CFU of M. bovis BCG/cp40 (G3), 106 CFU of M. bovis BCG/cp09720 (G4), M. bovis BCG/cp40 boosted with rCP40 (G5), and M. bovis BCG/cp09720 boosted with rCP09720 (G6). The highest survival rate of 90% was observed in the G5 group, followed by 80% in the G6 group and 70% in the G3 and G4 groups. Moreover, a significantly greater induction of IFN-γ and IL-10 was found in the G3 group and higher IL-17 levels were recorded in the G5 group compared to their levels in the control group (G1) (p < 0.05). A specific humoral immune response (total IgG) was found in the G5 and G6 groups on day 42 compared to the level of response in the G1 group. These results indicated that the vector vaccine elicited significantly greater survival of mice in all experimental groups after a strong virulent challenge and induced a strong immune response.

Bioprospection of the trichomonacidal activity of lipid extracts derived from marine macroalgae Gigartina skottsbergii.

Algal extracts are sources of bioactive substances with applications in the development of novel alternative drugs against several diseases, including trichomoniasis sexually transmitted infection caused by Trichomonas vaginalis. Factors such as clinical failures and resistant strains limit the success of the existing drugs available for treating this disease. Therefore, searching for viable alternatives to these drugs is essential for the treatment of this disease. The present study was conducted for, in vitro and in silico characterization of extracts obtained from marine macroalgae Gigartina skottsbergii at stages gametophidic, cystocarpic, and tetrasporophidic. In addition, antiparasitic activity of these extracts against the ATCC 30236 isolate of T. vaginalis, their cytotoxicity, and gene expression of trophozoites after treatment were evaluated. The minimum inhibitory concentration and 50% inhibition concentration were determined for each extract. Results: In vitro analysis of the extracts' anti-T. vaginalis activity revealed an inhibitory effect of 100%, 89.61%, and 86.95% for Gigartina skottsbergii at stages gametophidic, cystocarpic, and tetrasporophidic, respectively, at 100 µg/mL. In silico analysis revealed the interactions between constituents of the extracts and enzymes from T. vaginalis, with significant free energy values obtained for the binding. None of the extract concentrations exhibited cytotoxic effects on VERO cell line compared to control, while cytotoxicity on HMVII vaginal epithelial cells line was observed at 100 µg/mL (30% inhibition). Gene expression analysis revealed differences in the expression profile of T. vaginalis enzymes between the extract-treated and control groups. According to these results, Gigartina skottsbergii extracts exhibited satisfactory antiparasitic activity.

Evaluation of the Association of Recombinant Proteins NanH and PknG from Corynebacterium pseudotuberculosis Using Different Adjuvants as a Recombinant Vaccine in Mice.

Caseous lymphadenitis is a chronic contagious disease that causes economic losses worldwide. Treatments are ineffective, thus demonstrating the importance of vaccination. In this study, rNanH and rPknG proteins from Corynebacterium pseudotuberculosis were associated with saponin or aluminum hydroxide adjuvants. Three experimental groups (10 animals each) were immunized with sterile 0.9% saline solution (G1), rNanH + rPknG + Saponin (G2), rNanH + rPknG + Al(OH)3 (G3). The mice received two vaccine doses 21 days apart. Animals were challenged 21 days after the last immunization and evaluated for 50 days, with endpoint criteria applied when needed. The total IgG production levels of the experimental groups increased significantly on day 42 when compared to the control (p < 0.05). When tested against rNanH, G2 had a better rate of anti-rNanH antibodies compared to G3. In the anti-rPknG ELISA, the levels of total IgG, IgG1, and IgG2a antibodies were higher in G2. The vaccines generated partial protection, with 40% of the animals surviving the challenge. The association of recombinant NanH and PknG proteins led to promising protection rates in mice, and although using different adjuvants did not interfere with the survival rate, it influenced the immune response generated by the vaccine formulations.

In vitro nematocidal activity of Punica granatum L. against gastrointestinal helminths in goats.

The objective of this study was to evaluate the in vitro ovicidal activity, phytochemistry, and toxicity of a saline extract obtained from peel of Punica granatum L fruits. The ovicidal activity was evaluated by the hatching inhibition of eggs recovered from fecal samples of naturally infected goats; the phytochemical analysis was carried out using the fruit peel; and the toxicity was tested on Artemia salina, using saline extract. The results showed that the ovicidal effect of the tested extract was 99% (25 mg mL-1), 99% (12.5 mg mL-1), 98% (6.25 mg mL-1), and 95% (3.12 mg mL-1), higher than that of the control drug, thiabendazole (83%). The phytochemical analysis showed presence of phenols, anthraquinones, and condensed and hydrolysable tannins in the fruit extract. The toxicity test of the extract of P. granatum showed an LC50 of 6.19 mg mL-1, which indicates a safe use for a concentration of 3.12 mg mL-1, since it was the tested concentration that was below the reliable LC50. The saline extract from peels of P. granatum has ovicidal activity, important secondary metabolites, and absence of toxicity at the lowest concentration tested. However, in vivo tests in experimental models are recommended before performing experiments in ruminants.

Investigação dos perigos de natureza biológica em saladas comercializadas em restaurantes self service / Investigation of biological hazards in salads marketed in self service restaurants

Com o intuito de obter uma alimentação mais prática, que não interfira no cumprimento da jornada de trabalho, grande parcela dos indivíduos tem recorrido a refeições provenientes de estabelecimentos de autosserviço, inclusive as saladas prontas para o consumo. Neste sentido, este estudo teve por finalidade a investigação dos perigos de natureza biológica em saladas cruas de restaurantes tipo self service localizados no município de Mossoró, Rio Grande do Norte. Foram realizadas análises parasitárias, pesquisa de coliformes e determinação de Escherichia coli e Salmonella sp., observando-se a conformidade das amostras com a legislação vigente. Sobre os micro-organismos, foi constatada a presença de coliformes a 45°C, mas com valores dentro do preconizado, bem como a ausência de E. coli e Salmonella sp. Apesar de não detectada a presença de parasitos, foram observadas sujidades em algumas das saladas avaliadas. Tais resultados ressaltam a necessidade de maiores cuidados quanto à manipulação do produto.

In order to obtain a more practical diet, which does not interfere with the work day, a large proportion of individuals have used meals from self-service establishments, including ready-to-eat salads. In this sense, the purpose of the study was to investigate biological hazards in raw salads of self-service restaurants located in the municipality of Mossoró, Rio Grande do Norte. Parasite analyzes, coliform research and determination of Escherichia coli and Salmonella sp. were realized, observing the compliance of the samples with the current legislation. On the microorganisms, the presence of coliforms at 45°C, but with values within the recommended, as well as the absence of E. coli and Salmonella sp. Although no parasites were detected, soils were observed in some of the evaluated salads. These results highlight the need for greater care in handling the product.

Anthelmintic effect of Cassia fistula and Combretum leprosum protein fractions against goat gastrointestinal nematodes.

In this study, we evaluated the ovicidal and larvicidal activity of protein preparations obtained from Cassia fistula L. and Combretum leprosum Mart. leaves on the gastrointestinal parasites of goats. Protein preparations were obtained after the extraction of C. fistula L. and C. leprosum Mart. leaves, followed by protein fractionation (with ammonium sulfate saturation percentages of 30%, 30%-60%, and 60%-90%) and dialysis, which resulted in protein fractions (called F1, F2, and F3, respectively). The fractions were evaluated by egg hatching (the eggs were recovered in stool samples from naturally infected goats) and larval development tests. The results reveled that the inhibition of hatching of eggs caused by the protein fractions of C. fistula (38%) were similar to that of the control drug, thiabendazole. In addition, the fractions of C. fistula caused significant inhibition (61-69%) of larval development also. However, C. leprosum did not reveal significant inhibition of egg hatching and larval development. We conclude that C. fistula L. showed better ovicidal and larvicidal activity against endoparasites.

Anthelmintic effect of Cassia fistula and Combretum leprosum protein fractions against goat gastrointestinal nematodes / Efeito antihelmíntico das frações proteicas de Cassia fistula e Combretum leprosum contra nematodeos gastrintestinais de caprinos

Abstract In this study, we evaluated the ovicidal and larvicidal activity of protein preparations obtained from Cassia fistula L. and Combretum leprosum Mart. leaves on the gastrointestinal parasites of goats. Protein preparations were obtained after the extraction of C. fistula L. and C. leprosum Mart. leaves, followed by protein fractionation (with ammonium sulfate saturation percentages of 30%, 30%-60%, and 60%-90%) and dialysis, which resulted in protein fractions (called F1, F2, and F3, respectively). The fractions were evaluated by egg hatching (the eggs were recovered in stool samples from naturally infected goats) and larval development tests. The results reveled that the inhibition of hatching of eggs caused by the protein fractions of C. fistula (38%) were similar to that of the control drug, thiabendazole. In addition, the fractions of C. fistula caused significant inhibition (61-69%) of larval development also. However, C. leprosum did not reveal significant inhibition of egg hatching and larval development. We conclude that C. fistula L. showed better ovicidal and larvicidal activity against endoparasites.

Resumo Neste estudo, foram avaliadas as atividades ovicida e larvicida de preparações proteicas de Cassia fistula L. e Combretum leprosum Mart. em parasitas gastrointestinais de caprinos. As preparações proteicas foram obtidas por extração das folhas de C. fistula L. e C. leprosum Mart. seguido pelo fracionamento proteico (com porcentagens de saturação de sulfato de amônio de 30%, 30-60%, 60-90%) e diálise, resultando nas frações proteicas (intituladas F1, F2 e F3, respectivamente). As frações foram avaliadas nos testes de eclosão de ovos (os ovos foram recuperados em amostras de fezes de cabras naturalmente infectadas) e de desenvolvimento larvar. Os resultados revelaram que a inibição da eclosão de ovos causada pelas frações proteicas de C. fistula (38%) foi semelhante à do fármaco controle, o tiabendazol. Além disso, as frações de C. fistula também causaram inibição significativa (61-69%) do desenvolvimento larvar. No entanto, C. leprosum não revelou inibição significativa na eclosão dos ovos e no desenvolvimento larvar. Concluiu-se que C. fistula L. mostrou uma melhor atividade ovicida e larvicida contra endoparasitas.