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Aspergillus ochraceus and Aspergillus niger are spoilage and mycotoxin-producing fungi that can contaminate agricultural commodities and derived products. In the present study, menthol, eugenol, and their combination (mix 1:1) were tested to determine their contact and fumigation toxicity against the two fungi. Menthol, eugenol, and their mixture significantly reduced mycelial growth and spore germination at concentrations from 300 to 600 µg/mL, and the inhibitory effects showed clear dose dependence. The minimum inhibitory concentration (MIC) values against A. ochraceus were 500 µg/mL (menthol), 400 µg/mL (eugenol), and 300 µg/mL (mix 1:1), while the MIC values for A. niger were 500 µg/mL (menthol), 600 µg/mL (eugenol), and 400 µg/mL (mix 1:1). Additionally, the analyzed compounds exhibited more than 50% protection against A. ochraceus and A. niger by fumigation of stored cereal grains (maize, barley, and rice) in sealed containers. The binary mixture of menthol and eugenol showed synergistic effects against both fungi in both in vitro direct contact and stored grain fumigation trials. The results of the present study provide a scientific basis for the application of a combination of natural antifungals in food preservation.

New bioactive natural products, the phenolic composition and the biological activities of organic extracts from the needles of the Algerian Pinus coulteri were investigated. The analysis by HPLC-DAD of crude extract revealed the presence of 10 phenolic acids and nine flavonoids. In vitro anti-oxidant activities were performed using four different tests. The greatest antiradical activity was found in the ethyl acetate fraction (IC50 = 3.2 ± 0.3 µg/mL), whereas the diethyl ether fraction had the higher contents of total phenolics and flavonoids and exhibited a highest activity in reducing power and β-carotene-linoleic acid tests with EC50= 67.1 ± 0.4 μg/mL and 71.5 ± 0.2% of inhibition, respectively. Furthermore, a low to moderate antimicrobial activity according to all extracts was revealed against eight bacteria tested. The MIC value of chloroform fraction showed a strong degree of antibacterial activity (<0.09 mg/mL). The crude extract was found toxic with LC50 value of 15.2 μg/mL by brine shrimp toxicity assay. The needle extract of P. coulteri is rich in valuable biologically active compounds and could represent a new resource of anti-oxidant agents for the treatment of diseases.

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