Antagonistic lactic acid bacteria in association with Saccharomyces cerevisiae as starter cultures for standardization of sour cassava starch production.

In order to improve cassava's palatability and reduce its toxicity, this root is fermented and applied in foods, such as sour cassava starch used to prepare cheese bread and biscuits. This fermentation occurs spontaneously with lactic acid bacteria (LAB) and yeasts. However, it remains an empirical process, with long duration and lack of product quality homogeneity. This work aims to use starter cultures in a pilot-scale fermentation process for the production of sour cassava starch. After differentiation of strains, Lactobacillus plantarum Lp3, which exhibited great total titratable acidity (TTA) (5.01 ± 0.05%) and antagonistic activity against Bacillus cereus, Escherichia coli and Salmonella Typhimurium, together with Lactobacillus brevis Lb9 (with lesser TTA values: 2.71 ± 0.10%, but amylolytic activity: 2.75 ± 0.61 mm) were tested as single and co-cultures with Saccharomyces cerevisiae UFMG-A1007. LAB and yeasts were inoculated at counts of 8 and 7 log10 CFU/g, respectively, and they remained until the 28th day only in co-culture, highlighting the importance of the yeast for the LAB viability. Although single cultures lead to higher acidity during fermentation, the final product acidity obtained with single cultures did not differ from the acidity obtained with L. plantarum Lp3 in association with S. cerevisiae UFMG-A1007. Therefore, this co-culture exhibited higher potential to be tested as a starter culture in industrial-scale fermentation studies because both microorganisms were in high counts until the end of fermentation and contributed to a final product safe for human consumption, with satisfactory acidity, expansion capacity, and physicochemical properties.

Synthesis, molecular modeling studies and evaluation of antifungal activity of a novel series of thiazole derivatives.

In the search for new antifungal agents, a novel series of fifteen hydrazine-thiazole derivatives was synthesized and assayed in vitro against six clinically important Candida and Cryptococcus species and Paracoccidioides brasiliensis. Eight compounds showed promising antifungal activity with minimum inhibitory concentration (MIC) values ranging from 0.45 to 31.2 µM, some of them being equally or more active than the drug fluconazole and amphotericin B. Active compounds were additionally tested for toxicity against human embryonic kidney (HEK-293) cells and none of them exhibited significant cytotoxicity, indicating high selectivity. Molecular modeling studies results corroborated experimental SAR results, suggesting their use in the design of new antifungal agents.

Antifungal activity of eicosanoic acids isolated from the endophytic fungus Mycosphaerella sp. against Cryptococcus neoformans and C. gattii.

The antifungal effects of two eicosanoic acids, 2-amino-3,4-dihydroxy-2-25-(hydroxymethyl)-14-oxo-6,12-eicosenoic acid (compound 1) and myriocin (compound 2), isolated from Mycosphaerella sp. were evaluated against Cryptococcus neoformans and C. gattii. The compounds displayed antifungal activities against several isolates of C. neoformans and C. gattii, with minimal inhibitory concentration (MIC) values ranging from 0.49 to 7.82 µM for compound 1 and 0.48-1.95 µM for compound 2. In the checkerboard microtiter test, both compounds exhibited synergistic activity with amphotericin B against C. gattii. Ultrastructural analysis revealed several signs of damage in C. gattii and C. neoformans cells treated with compounds 1 and 2, including deformities in cell shape, depressions on the surface, and withered cells. The cells of C. gattii treated with compounds 1 and 2 showed less loss of cellular material in comparison to those treated with amphotericin B. The difference in cellular material loss increased in a test compound concentration-dependent manner. Consistent with this observation, compounds 1 and 2 were able to internalize propidium iodide (PI) in C. gattii cells. In addition, compound 2 induced the formation of several pseudohyphae, suggesting that it could reduce virulence in C. gattii cells. The study results show that these natural products led to membrane damage; however, this may not be the main target of action. These compounds have potential antifungal activity and could be useful in further studies for developing more effective combination therapies with amphotericin B and reducing side effects in patients.

Thiazole compounds with activity against Cryptococcus gattii and Cryptococcus neoformans in vitro.

Human cryptococcosis can occur as a primary or opportunistic infection and develop as an acute, subacute, or chronic, systemic infection involving different host organs. We evaluated the antifungal activity of thirteen compounds against Cryptococcus gattii and Cryptococcus neoformans in vitro, by assessing the toxicity of the compounds showing the greatest antifungal activity in VERO cells and murine macrophages. From these results, four compounds were considered promising for further studies because they displayed low cytotoxicity and significant antifungal activity. The heterocyclic compounds 1b, 1c, 1d, and 1m have antifungal activity levels between that of amphotericin B and fluconazole in vitro. The death curve of Cryptococcus spp. treated with these four compounds was similar to the curve obtained for amphotericin B, in that we observed a significant reduction in cell viability within the first 24 h of treatment. Additionally, we found that there was no effect when these compounds were combined with amphotericin and fluconazole, except for 1c, which antagonized the effect of amphotericin B against C. gattii, also reflected in the reduction of the post-antifungal effect (PAFE); however, this interaction did not alter the ergosterol content. The results shown in this paper reveal the discovery of novel thiazole compounds, which are easy to synthesize, and with potentially exhibit antifungal activity, and display low cytotoxicity in normal mammalian cells. These compounds can be used as prototypes for the design of new antifungal drugs against C. gattii and C. neoformans.