Inclusion Complexes of Litsea cubeba (Lour.) Pers Essential Oil into ß-Cyclodextrin: Preparation, Physicochemical Characterization, Cytotoxicity and Antifungal Activity.

The uses of natural compounds, such as essential oils (EOs), are limited due to their instability to light, oxygen and temperature, factors that affect their application. Therefore, improving stability becomes necessary. The objective of this study was to prepare inclusion complexes of Litsea cubeba essential oil (LCEO) with ß-cyclodextrin (ß-CD) using physical mixing (PM), kneading (KN) and co-precipitation (CP) methods and to evaluate the efficiency of the complexes and their physicochemical properties using ATR-FTIR, FT-Raman, DSC and TG. The study also assessed cytotoxicity against human colorectal and cervical cancer cells and antifungal activity against Aspergillus flavus and Fusarium verticillioides. The complexation efficiency results presented significant evidence of LCEO:ß-CD inclusion complex formation, with KN (83%) and CP (73%) being the best methods used in this study. All tested LCEO:ß-CD inclusion complexes exhibited toxicity to HT-29 cells. Although the cytotoxic effect was less pronounced in HeLa tumor cells, LCEO-KN was more active against Hela than non-tumor cells. LCEO-KN and LCEO-CP inclusion complexes were efficient against both toxigenic fungi, A. flavus and F. verticillioides. Therefore, the molecular inclusion of LCEO into ß-CD was successful, as well as the preliminary biological results, evidencing that the ß-CD inclusion process may be a viable alternative to facilitate and increase future applications of this EO as therapeutic medication, food additive and natural antifungal agent.

Analytical and toxicological aspects of dithiocarbamates: an overview of the last 10 years.

Compilation studies related to toxicological aspects and also biological monitoring and analysis methods for specific fungicides and, mainly, those that belong to the class of the dithiocarbamates (DTCs) have not been carried out at least in the last ten years. DTCs - dimethyldithiocarbamates, ethylenebisditiocarbamates, propylenebisditiocarbamates - are organosulfur compounds that form complexes due to the presence of different chemical elements, which bind strongly and inhibit enzymes that are essential to the functioning of the organism, causing a serious proven adverse effect on biological systems, such as alteration of thyroid hormones, teratogenesis and neurotoxicity. It is still evident, as shown by world data, that the growing consumption of fungicides has increasingly exposed the population in general and, in particular, workers who deal with these substances. There is a scarcity of studies in the literature discussing the toxicological and analytical aspects that are important for understanding the real effects of DTCs and monitoring human exposure to them. Therefore, the aim of this work was to expose, in a comprehensive way and through a narrative review, gaps in research related to the fungicides of the DTCs class, their metabolites, as well as the toxicological and analytical aspects involved. The review is divided into two parts: (1) Toxicological aspects, including toxicokinetics, toxicodynamics and toxidromes; and (2) Analytical Toxicology, which comprises biomarkers, sample preparation and identification/quantification methods.

Anti-mycotoxigenic and antifungal activity of ginger, turmeric, thyme and rosemary essential oils in deoxynivalenol (DON) and zearalenone (ZEA) producing Fusarium graminearum.

This study aimed to evaluate the antimycotoxigenic effect of essential oils (EOs) obtained from four different aromatic plants on the production of deoxynivalenol (DON) and zearalenone (ZEA) by Fusarium graminearum. The EOs from ginger (GEO), turmeric (TEO), thyme (ThEO) and rosemary (REO) were obtained by hydrodistillation and identified by gas chromatography/mass spectrometry (GC/MS). The major compounds found were mostly monoterpenes and sesquiterpenes. The minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were 11.25, 364, 366 and 11,580 µg mL-1 for ThEO, GEO, REO and TEO, respectively. The results evidenced that the assessed EOs inhibited DON and partially ZEA production by F. graminearum. ThEO and GEO were the EOs with most potent antimycotoxigenic action for DON and ZEA, respectively. These EOs have shown promising results in vitro regarding inhibition of mycotoxin production and might be used in the future as substitutes for synthetic fungicides.

Metals in Brazilian family farming grapes and estimated daily intake.

The aim of this study was to determine concentrations of metals in peel, pulp, and seeds of grapes obtained from family farms in Brazil, compare them to the maximum threshold levels and to evaluate the risk by estimating the daily intake (EDI). Grape samples were collected from farms and levels of Cd, Cr, Cu, Mn, Ni, Pb and Zn were assessed via ICP-MS. The highest metal levels were found in grape peels, Cu at the highest concentration (107.6 mg kg-1). Cr, Cu, and Pb were found at concentrations which exceeded maximum threshold levels. The EDI of Cd, Cu and Pb through consumption of grapes for the assessed Brazilian population was 0.29, 1822 and 3.02 µg/kg bw/day, respectively. The EDI of Cu was above the Provisionary Tolerable Daily Intake (PTDI). Thus, there are possible health risks due to the occurrence of Cu in Brazilian grapes.

Litsea cubeba essential oil: chemical profile, antioxidant activity, cytotoxicity, effect against Fusarium verticillioides and fumonisins production.

The purpose of this study was to determine the chemical profile of Litsea cubeba essential oil, carry out an in vitro evaluation of its antioxidant potential and its cytotoxicity, as well as its antifungal and antimicotoxigenic activities against Fusarium verticillioides. Most of the compounds observed in the EO were neral (32.75%) and geranial (37.67%). The radical scavenging capacity of 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid was 104.4 and 56.4 mmol Trolox mg-1, respectively, indicating good antioxidant activity. The EO studied by us revealed cytotoxic effect against HT-29 and HeLa cancer cells. The Minimum Inhibitory and Minimum Fungicidal Concentrations against F. verticillioides were both 125 µg mL-1. Morphological investigation, performed by fluorescence microscopy and scanning electron microscopy, showed that hyphae and microconidia structures underwent changes after treatment with the EO. Analyses performed with the EO strongly reduced the mycelial development of F. verticillioides and the synthesis of fumonisins B1 and B2 in dose-dependence effect compared (P < 0.01) with the fungal control (105 conidia mL-1) and positive control (fludioxonil + metalaxyl-M). Thus, the results obtained in vitro suggest that L. cubeba EO has excellent antioxidant, fungicidal, and antimycotoxigenic effects.

Occurrence, exposure evaluation and risk assessment in child population for aflatoxin M1 in dairy products in Brazil.

This study aimed to evaluate the risk concerning child population's health because of the occurrence of AFM1 in UHT milk, powdered milk (PM) and infant formulae (IF). Determination of AFM1 was performed in 60 samples and evaluation of the mycotoxin exposure was carried out through the determination of the estimated daily intake (EDI), whereas risk characterization was evaluated with the calculation of the risk of Hepatocellular Carcinoma (HCC) and the Margin of Exposure (MOE). AFM1 ranged from 150 to 1020 ng/kg, and all the positive samples exceeded the limits stablished by European Community. The EDI for AFM1 ranged according to the age group of the population studied (0-5 years old) from 0.828 to 2.523, 0-2.113 and 0.029-0.833 ng/kg b. w./day in UHT, PM and IF, respectively. The number of HCC cases associated with AFM1 exposure (0.0015 a 0.0045) was higher than the limit of 0.001 case/100,000. MOE values for AFM1 were 728 to 239, considerably below the security margin of 10,000. These results point to a potential risk to the health of Brazilian child population exposed to AFM1 in dairy products.

Antifungal and antimycotoxigenic effects of Zingiber officinale, Cinnamomum zeylanicum and Cymbopogon martinii essential oils against Fusarium verticillioides.

There is an increasing demand for fungi control in grains, especially toxigenic. Also, there is growing concern on the use of synthetic fungicides; thus alternatives are needed. The aim of this study was to evaluate the antifungal and antimycotoxigenic action of essential oils (EOs) from Zingiber officinale, Cinnamomum zeylanicum and Cymbopogon martinii against Fusarium verticillioides, a spoilage and toxigenic fungus. Essential oils were first chemically characterised by gas chromatography coupled to mass spectrometry, and their antioxidant potential was measured by the DPPH, ABTS and FRAP methods. Minimum inhibitory concentration (MIC) and disc diffusion were used to assess antifungal activity. Scanning electron microscopy was used to evaluate morphological changes in the fungus. Antimycotoxigenic activity of the EOs against the production of fumonisin B1 and B2 by F. verticillioides was evaluated using ultra-high-performance liquid chromatography system. Z. officinale, C. zeylanicum and C. martinii EOs were predominantly composed by zingiberene and geranial; eugenol; and geraniol, respectively. All the EOs had high antioxidant power, especially that from C. zeylanicum. The MICs were 250, 500 and 2,000 µg mL-1 for C. zeylanicum, C. martinii and Z. officinale EOs, respectively. Mycelial reduction of F. verticillioides was observed when EOs were used, and the lowest activity was detected in the Z. officinale EO. Overall, the tested EOs promoted structural damage to the fungal cell wall, decreased conidia size and mycelial reduction. Antimycotoxigenic evaluation of the EOs evidenced a significant reduction (p < .05) in the production of fumonisins B1 and B2 with all the EOs evaluated in the study. These results suggest that especially C. zeylanicum and C. martinii EOs are highly useful for controlling F. verticillioides and fumonisins production.

Occurrence and risk assessment of population exposed to deoxynivalenol in foods derived from wheat flour in Brazil.

Deoxynivalenol (DON) is the most important of the trichothecenes in terms of amounts and occurrence in wheat. This compound was shown to be associated with a glomerulonephropathy involving an increase of immunoglobulin A in humans. This study assessed the occurrence of DON in wheat flour and the exposure of Brazilian teenagers, adults and elderly to this mycotoxin due to intake of wheat flour-based products. DON extraction in wheat flour was carried out by solid phase extraction and the quantification was performed by ultra-high proficiency liquid chromatography with diode-array detection. A total of 77.9% of all samples were positive for DON, with concentrations ranging from 73.50 to 2794.63 µg kg-1. The intake was calculated for the average and 90th percentile of the contamination levels of DON in foods based-wheat for teenagers, adults and elderly in Brazil, and compared with the provisional maximum tolerable daily intakes (PMTDI). Females of all age groups were exposed to DON at higher levels when compared to males in regard of consumption of breads and pastas. Teenagers were the main consumers of foods derived from wheat flour, with maximum probable daily intakes of 1.28 and 1.20 µg kg-1 b.w. day-1 for females and males, respectively. This population is at an increased risk of exposure to DON due to consumption of wheat flour-based foods in Brazil.