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.

Efeitos antifúngico e antiaflatoxigênico de timol e carvacrol em Aspergillus flavus / Antifungal and antiaflatoxigenic activities of thymol and carvacrol against Aspergillus flavus

O objetivo deste estudo foi avaliar os efeitos do timol e carvacrol contra Aspergillus flavus e a produção de aflatoxinas. Foram determinadas a concentração inibitória mínima (CIM), o espectro fungitóxico e a atividade inibidora de micotoxinas pelo timol e carvacrol. Os resultados mostraram que timol e carvacrol exibiram ação fungicida de acordo com a CIM de 2500 e 30 µg mL-1, respectivamente. Tanto o timol quanto o carvacrol inibiram significativamente o crescimento de A. flavus (p<0,05) a partir de 600 e 15 µg mL-1, respectivamente. A biomassa fúngica, estimada pela determinação da concentração de ergosterol, foi significativamente reduzida (p <0,05) em 2500 µg mL-1 de timol e 250 µg mL-1 de carvacrol. O timol e o carvacrol exibiram efeitos antiaflatoxigênicos em 600 e 125 µg mL-1, respectivamente. O timol e o carvacrol exibiram atividade antifúngica, mas não exibiram alta atividade antiaflatoxigênica. Carvacrol e timol podem ser considerados como potentes compostos naturais antifúngicos contra A. flavus.

The aim of this study was to evaluate the effects of thymol and carvacrol upon production of aflatoxins by Aspergillus flavus and upon its growth. Minimal inhibitory concentration (MIC), fungitoxic spectrum and mycotoxin inhibition were assessed. Results showed thymol and carvacrol exhibited fungicidal action, as determined by MIC values of 2500 and 30 µg mL-1, respectively. Both thymol and carvacrol significantly inhibited growth of A. flavus (p<0.05) at concentrations of 600 and 15 µg mL-1, respectively. Fungal biomass, as estimated by determination of ergosterol concentration, was significantly reduced (p<0.05) at thymol concentrations of 2500 µg mL-1 and at carvacrol concentrations of 250 µg mL-1. Thymol and carvacrol exhibited antiaflatoxigenic effects at concentrations of 600 and 125 µg mL-1, respectively. While both thymol and carvacrol showed possessing antifungal activities, neither were highly antiaflatoxigenic. Carvacrol and thymol might be considered for use as potential antifungal natural compounds against A. flavus.

Antifungal activity and inhibition of aflatoxins production by Zingiber officinale Roscoe essential oil against Aspergillus flavus in stored maize grains / Atividade antifúngica e inibição da produção de aflatoxinas pelo óleo essencial de Zingiber officinale Roscoe contra Aspergillus flavus em grãos de milho armazenados

ABSTRACT: Essential oils are possible alternatives to the use of synthetic pesticides for control of fungal contamination. Ginger (Zingiber officinale) essential oil (GEO) is known for having antifungal and antiaflatoxigenic properties, but its use as a fumigant in situ has not been studied yet. The aim of this study was to evaluate GEO's effects upon Aspergillus flavus as a fumigant agent in stored maize grains. The main compounds reported in GEO were α-zingiberene (23.85%) and geranial (14.16%), characterized by gas chromatography-mass spectrometry and nuclear magnetic resonance. The GEO was used as a fumigant in irradiated maize grains in concentrations ranging from 5 to 50 µg/g and the resulting effects were compared to a synthetic antifungal agent (carbendazim and thiram), an antifungal traditionally used for seed treatment. The antifungal efficacy of GEO against A. flavus has been proven in a dose-dependent manner through in situ (maize grains) test. The GEO inhibited aflatoxin production at concentrations 25 and 50 µg/g and controlled fungal growth. Therefore, GEO can be used as an effective and non-toxic alternative to conventional treatments in stored maize grains for the natural control of A. flavus.

RESUMO: Os óleos essenciais são possíveis alternativas ao uso de praguicidas sintéticos no controle da contaminação por fungos. O óleo essencial de gengibre (Zingiber officinale - OEG) é conhecido por possuir propriedades antifúngicas e antiaflatoxigênicas, mas seu uso como fumigante in situ ainda não foi estudado. O objetivo deste estudo foi avaliar os efeitos do OEG sobre Aspergillus flavus como agente fumigante em grãos de milho armazenados. Os principais compostos encontrados no OEG foram α-zingibereno (23,85%) e geranial (14,16%), caracterizados por cromatografia gasosa acoplada a espectrometria de massas e ressonância magnética nuclear. O OEG foi utilizado como fumigante em grãos de milho irradiados em concentrações variando de 5 a 50 µg/g e os efeitos resultantes foram comparados a um agente antifúngico sintético (carbendazim e tiram), antifúngico tradicionalmente usado para o tratamento de sementes. A eficácia antifúngica do OEG contra A. flavus foi comprovada de maneira dependente da dose através do teste in situ (grãos de milho). O OEG inibiu a produção de aflatoxina nas concentrações 25 e 50 µg/g e controlou o crescimento de fungos. Portanto, o OEG pode ser usado como uma alternativa eficaz e não tóxica aos tratamentos convencionais em grãos de milho armazenados para o controle natural de A. flavus.