Effect of the essential oils from Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on the synthesis of ochratoxin A by Aspergillus niger and Aspergillus carbonarius isolated from tropical wine grapes.

The influence of essential oils (EOs) extracted from the leaves of Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on ochratoxin A (OTA) synthesis by fungi was studied. The extraction of EOs was performed by hydrodistillation (Clevenger apparatus) over a 2-h period and subsequently analyzed by GC-MS and GC-FID. The toxigenic activity of the essential oils (31.25; 15.62 and 7.81 µg mL-1) was evaluated by inhibiting the production of OTA by Aspergillus niger and Aspergillus carbonarius in Czapek agar medium culture. The quantification of the toxin was performed by HPLC. The production of OTA was dependent on the fungal species, incubation temperature (15 and 25 °C) and the presence of the essential oils. In tests carried out at 15 °C, the oils caused a reduction in OTA synthesis that ranged from 57.60 to 76.93% and from 54.78 to 98.68% for the fungal species A. carbonarius and A. niger, respectively. At 25 °C, reductions ranged from the 38.66 to 75.93% and from 17.94 to 71.79% for the respective fungi. The study concluded that natural products could be potential biocontrol agents against OTA contamination in food.

Influence of physical and chemical characteristics of wine grapes on the incidence of Penicillium and Aspergillus fungi in grapes and ochratoxin A in wines.

The incidence of filamentous fungi and toxin levels in grapes and wines varies depending on the variety of grapes, the wine region, agricultural practices, weather conditions, the harvest and the winemaking process. In this sense, the objective of this study was to evaluate the diversity of Aspergillus and Penicillium fungi isolated from wine grapes of the semi-arid tropical region of Brazil, evaluate the presence of ochratoxin A (OTA) in the experimental wine and verify if there is a correlation between occurrence of these fungi and the physicochemical characteristics of the wine grapes grown in the region. For the isolation of fungi we used the direct plating technique. The presence of OTA in the experimental wine was detected by high-performance liquid chromatography. The species found were Aspergillus niger, A. carbonarius, A. aculeatus, A. niger Aggregate, A. flavus, A. sojae, Penicillium sclerotiorum, P. citrinum, P. glabrum, P. decumbens, P. solitum and P. implicatum. All isolates of A. carbonarius were OTA producers and all P. citrinum were citrinin producers. The highest concentration of OTA was found in red wine (0.29µg/L). All species identified in this study, except A. flavus, showed a positive correlation with at least one physicochemical parameter assessed, highlighting the pectin content, total sugar, total acidity and phenolic compounds.

Effect of Fungicides on Growth and Ochratoxin A Production by Aspergillus carbonarius from Brazilian Wine Grapes.

Prevention in the field of mycotoxin-producing fungi is the most effective strategy for controlling the presence of mycotoxins in foods. Chemical fungicides are widely used to protect crops, so their implications on mycotoxin production need to be considered. Therefore, the aim of this study was to evaluate the effect in vitro and on grapes of five fungicides commonly used on grape cultures in Brazil on Aspergillus carbonarius growth and ochratoxin A (OTA) production. At the doses recommended by manufacturers, most fungicides significantly reduced A. carbonarius growth and OTA production in vitro, whereas this effect was influenced by the type of fungicide, dose, and temperature. Temperature was the main factor that influenced the effectiveness of fungicides. In general, at 15°C, fungicides showed the greatest reduction on fungal growth and OTA production. Fungicide effect on grapes was different to that on a semisynthetic grape medium. All fungicide doses were not effective at controlling A. carbonarius in grapes. Thus, the direct effect of fungicides on grapes must be studied to obtain a better approximation of field conditions. The results indicate that the use of fungicides at the doses recommended by manufacturers is better than the application at low doses. This study showed that at the lowest doses, where fungal growth is not inhibited, fungicides positively stimulate OTA production.

Effect of temperature, water activity, and pH on growth and production of ochratoxin A by Aspergillus niger and Aspergillus carbonarius from Brazilian grapes.

The growth of ochratoxigenic fungus and the presence of ochratoxin A (OTA) in grapes and their derivatives can be caused by a wide range of physical, chemical, and biological factors. The determination of interactions between these factors and fungal species from different climatic regions is important in designing models for minimizing the risk of OTA in wine and grape juice. This study evaluated the influence of temperature, water activity (aw), and pH on the development and production of OTA in a semisynthetic grape culture medium by Aspergillus carbonarius and Aspergillus niger strains. To analyze the growth conditions and production of OTA, an experimental design was conducted using response surface methodology as a tool to assess the effects of these abiotic variables on fungal behavior. A. carbonarius showed the highest growth at temperatures from 20 to 33°C, aw between 0.95 and 0.98, and pH levels between 5 and 6.5. Similarly, for A. niger, temperatures between 24 and 37°C, aw greater than 0.95, and pH levels between 4 and 6.5 were optimal. The greatest toxin concentrations for A. carbonarius and A. niger (10 µg/g and 7.0 µg/g, respectively) were found at 15°C, aw 0.99, and pH 5.35. The lowest pH was found to contribute to greater OTA production. These results show that the evaluated fungi are able to grow and produce OTA in a wide range of temperature, aw, and pH. However, the optimal conditions for toxin production are generally different from those optimal for fungal growth. The knowledge of optimal conditions for fungal growth and production of OTA, and of the stages of cultivation in which these conditions are optimal, allows a more precise assessment of the potential risk to health from consumption of products derived from grapes.

Synthesis, HIV-RT inhibitory activity and SAR of 1-benzyl-1H-1,2,3-triazole derivatives of carbohydrates.

This paper describes the synthesis of several 1-benzyl-1H-1,2,3-triazoles attached to different carbohydrate templates and their in vitro inhibitory profile against HIV-1 reverse transcriptase. In addition a theoretical comparison of the most active compounds with other classical antivirals was also performed. Our results showed 2a, 2d and 2g as the most active compounds that inhibited the HIV-1 reverse transcriptase catalytic activity with cytotoxicity lower than AZT and SI higher than DDC and DDI. The overall theoretical analysis of the molecular descriptors of 2a, 2d and 2g revealed that their HOMO energy is similar to other antivirals in use (AZT, DDC, DDI and 3TC) and together with the volume may contribute for the biological profile as they may allow new interactions with the target. In fact the 1,2,3-triazole compounds presented more lipophilicity and higher molecular volume and weight than the antivirals studied, which suggested that these features might not only contribute for new interactions with the HIV-RT but also influence the specificity and consequently the low cytoxicity profile of these compounds. Thus these data point them as promising leading compounds for generating new anti-HIV-RT compounds.

SAR of a series of anti-HSV-1 acridone derivatives, and a rational acridone-based design of a new anti-HSV-1 3H-benzo[b]pyrazolo[3,4-h]-1,6-naphthyridine series.

Herpes Simplex Virus (HSV) infections are among the most common human diseases. In this work, we assess the structural features and electronic properties of a series of ten 1-hydroxyacridone derivatives (1a-j) recently described as a new class of non-nucleoside inhibitors of Herpes Simplex Virus-1 (HSV-1). Based on these molecules, we applied rigid analogue and isosteric replacement approaches to design and synthesize nine new 3H-benzo[b]pyrazolo[3,4-h]-1,6-naphthyridine derivatives (2a-i). The biological and computational results of these new molecules were compared with 1-hydroxyacridones. An inhibitory profile was observed in 10-Cl substituted 3H-benzo[b]pyrazolo[3,4-h]-1,6-naphthyridine derivative (2f), which presents the same substituent at the analogous position of 1-hydroxyacridone derivative (1b). The structure-activity relationship (SAR) studies pointed out the 10-position next to nitrogen atom as important for the anti-HSV-1 profile in the pyrazolo-naphthyridine derivatives tested, which reinforced the promising profile for further experimental investigation. The most potent acridone and pyrazolo-naphthridine derivatives were also submitted to an in silico ADMET screening in order to determine their overall drug-score, which confirmed their potential antiviral profile.