Tihomir Kovač

Antifungal and antiaflatoxigenic activities of coumarinyl thiosemicarbazides against Aspergillus flavus NRRL 3251

Abstract The antifungal and antiaflatoxigenic effects of two series of coumarinyl thiosemicarbazides on Aspergillus flavus NRRL 3251 were studied. Fungi were grown in YES medium for 72 h at 29 °C in the presence of 0, 0.1, 1, and 10 μg mL-1 of coumarinyl thiosemicarbazides: one series with substitution in position 7 and another with substitution in position 4 of the coumarin core. Dry mycelia weight determination was used for antifungal activity estimation, while the aflatoxin B1 content in YES media, determined by the dilute and shoot LC-MS/MS technique, was used for the antiaflatoxigenic effect estimation. Standard biochemical assays were used for oxidative status marker (TBARS, SOD, CAT, and GPX) determination in A. flavus NRRL 3251 mycelia. Results show that 7-substituted-coumarinyl thiosemicarbazides possess a better antifungal and antiaflatoxigenic activity than 4-substituted ones. The most prominent substituted compound was the compound 3, N-(4-chlorophenyl)-2-(2-((4-methyl-2-oxo-2H-chromen-7-yl)oxy)acetyl)hydrazine-1-carbothioamide, which completely inhibited aflatoxin production at the concentration of 10 μg mL-1. Oxidative stress response of A. flavus exposed to the selected compounds points to the modulation of oxidative stress as a possible reason of aflatoxin production inhibition.

Inhibitory effect of coumarin derivatives on apple (cv. Idared) polyphenol oxidase

Inhibitory effect of 32 coumarin derivatives (20 Schiff bases, 5 thiosemicarbazides, 5 thiazolidinones, and their precursors, 7- hydroxy-4-methylcoumarin and 4-methylcoumarin- 7-yl hydrazine carboxylate) on partially purified apple polyphenol oxidase was investigated. Thirteen coumarin derivatives inhibited polyphenol oxidase (5 Schiff bases, 5 thiosemicarbazides, 1 thiazolidinone, 4-methyl- 7-hydroxycoumarin and 4-methylcoumarin-7-yl hydrazine carboxylate), while 19 derivatives showed no effect on enzyme activity. The most effective inhibitors were thiosemicarbazides, with 4-methyl-1-(2-(4-methyl-2-oxo-2H-chromen- 7-yloxy)acetyl) thiosemicarbazide (compound C23) being the most prominent inhibitor (IC50 = 10.45 µM). The importance of thiosemicarbazide moiety as crucial structure element for strong apple PPO inhibition was confirmed by its cyclisation to thiazolidinone bearing the same substituents as corresponding thiosemicarbazide. Capture of the sulphur atom of thiosemicarbazide group within tiazolidinone ring caused significant loss of inhibitory effect against apple PPO.

Fullerol C60(OH)24 nanoparticles modulate aflatoxin B1 biosynthesis in Aspergillus flavus

The water soluble fullerene C60 daughter product - fullerols C60(OH)24 (FNP) possesses a great potential of modifying secondary metabolites biosynthesis. In order to clarify the extent of interaction, the impact of FNP (10, 100 and 1000 ng mL−1) on aflatoxin production and the available precursors of biosynthesis pathway from Aspergillus flavus NRRL 3251 was determined, in both the mycelia and yeast extract sucrose (YES) medium, during a 168-hour growth period at 29 °C in the dark. The FNP of 8 nm in diameter, and with a zeta potential of −33 mV affected mycelial growth at 1000 ng mL−1 while conidia production was slightly affected at 10 ng mL−1. The FNP effect on aflatoxin and it biosynthetic precursors was concentration dependent and alteration of the sterigmatocystin (ST) export from the cell was observed. Most of the monitored aflatoxin precursors, except norsolorinic acid, were detected in both mycelia and YES medium. However, observed precursor concentrations were much higher in mycelia, with exception of ST. The study shows the loss of FNP antioxidative effect after 120 hours of growth, and strong concentration dependent aflatoxigenic effect after that time. Thus, this data is relevant to guide future considerations on FNP-fungal interactions in the environments and on risk assessment.

Jučer maskirani, a danas modificirani – što je sljedeće s mikotoksinima?

Abstract Mycotoxins are secondary metabolites produced by toxigenic fungi in crops worldwide. In (micro)organisms such as plants, fungi, bacteria, or animals they may be further metabolised and modified, but this is also true for food processing, which may lead to a wide range of masked mycotoxin forms. These often remain undetected by analytical methods and are the culprits for underestimates in risk assessments. Furthermore, once ingested, modified mycotoxins can convert back to their parent forms. This concern has raised the need for analytical methods that can detect and quantify modified mycotoxins as essential for accurate risk assessment. The promising answer is liquid chromatography-mass spectrometry. New masked mycotoxin forms are now successfully detected by iontrap, time-of-flight, or high-resolution orbitrap mass spectrometers. However, the toxicological relevance of modified mycotoxins has not been fully clarified.

Correction to: Characterization of Croatian Honeys by Right-Angle Fluorescence Spectroscopy and Chemometrics

The original version of this article unfortunately contained an incorrect version of author’s names and surnames (underlined). The correct names and surnames are: “Ivica Strelec”, “Lidija Brodar”, “Ivana Flanjak”, “Frane Čačić Kenjerić”, “Tihomir Kovač”, “Daniela Čačić Kenjerić”, “Ljiljana Primorac”.