Effect of treatment of Fusarium head blight infected barley grains with hop essential oil nanoemulsion on the quality and safety of malted barley.

Fusarium mycotoxin contamination of malting barley has been a persistent food safety issue for malting companies. In this study, the effect of hop essential oil (HEO) nanoemulsion on fungal biomass and mycotoxin production during the malting process was evaluated. Furthermore, the localization of fungal hyphae on the surface and inside the tissue of barley and malts was observed. The application of HEO nanoemulsion reduced fungal biomass and deoxynivalenol (DON) contents at each stage of the malting process as compared to control. During malting process, the fungal hyphae on kernel surfaces was reduced appreciably after steeping. However, the increment of hyphae was observed between the husk and testa layer of barley after germination than raw barley grains. In addition to its antifungal activity, the antioxidant activity of HEO in the treated malts suppressed the formation of aldehydes. This study lays the foundation for the utilization of HEO in the malting industry.

Clove oil-in-water nanoemulsion: Mitigates growth of Fusarium graminearum and trichothecene mycotoxin production during the malting of Fusarium infected barley.

Fusarium mycotoxin contamination in malting barley is of great concerns in malting industry. Our recent study found that clove oil nanoemulsions can act as highly efficient antifungal agents in vitro. Therefore, we explored the efficacy of clove oil nanoemulsions on Fusarium growth and mycotoxin during malting process. The impact of emulsifier types (Tween 80, BSA and quillaja saponins) on the formation of clove oil nanoemulsion, the mitigation effects on mycotoxin levels and fungal biomass, and the clove oil flavor residues on malts were measured. We observed that 1.5 mg clove oil/g nanoemulsion showed a negligible influence on germinative energy of barley, while still efficiently eliminated the DON levels and toxicogenic fungal biomass as quantified by Tri5 DNA content. Tween 80-stablized clove oil nanoemulsion displayed higher mycotoxin inhibitory activity and less flavor impact on the final malt. The results indicated the potential application of essential oil nanoemulsion during the malting process.

Influence of nonionic and ionic surfactants on the antifungal and mycotoxin inhibitory efficacy of cinnamon oil nanoemulsions.

The influence of ionic surfactants (cationic surfactant lauric arginate and anionic surfactant lysolecithin) on the physical properties, antifungal and mycotoxin inhibitory efficacy of Tween 80 stabilized cinnamon oil-in-water nanoemulsions was investigated. Nanoemulsion droplets of similar particle diameter (∼100 nm), but variable electrical characteristics, were formed by mixing 0.1 wt% ionic surfactant with 0.9 wt% Tween 80 before homogenization. The nanoemulsions were physically stable over 28 days at 23 °C. The antifungal activity (against mycelial growth and spore germination) and mycotoxin inhibitory activity of cinnamon oil nanoemulsions bearing positive, neutral, and negative charge surface was then evaluated against two chemotypes of Fusarium graminearum. In general, the cinnamon oil played a decisive role in the resulting antifungal and mycotoxin inhibitory activities. The surfactant charge had a limited impact on the antifungal mycotoxin inhibitory activities of cinnamon oil in the nanoemulsions. Both ionic surfactant-based cinnamon oil nanoemulsions showed greater activity in inhibiting mycelial growth and mycotoxin production of F. graminearum than those based on Tween 80. Treatment of mycelium with cinnamon oil nanoemulsions resulted in the loss of cytoplasm from fungal hyphae, and accounted for the antifungal action. These results have important implications for the design of essential oil based nanoemulsions as effective antifungal delivery systems in foods.

Influence of oil phase composition on the antifungal and mycotoxin inhibitory activity of clove oil nanoemulsions.

The influence of oil composition on the physical properties and antifungal and mycotoxin inhibitory activity of clove oil-in-water nanoemulsions was investigated. Physically stable clove oil-in-water nanoemulsions could be fabricated by incorporating either ≥75 wt% of corn oil or ≥50 wt% of medium chain triacylglycerol (MCT) into clove oil before homogenization to prevent Ostwald ripening. The clove oil-in-water nanoemulsions with mean diameters of <150 nm showed high physical stability over 30 days storage. The antifungal activity of physically stable clove oil nanoemulsions was further evaluated using effective concentration (EC) and inhibitory activity towards mycotoxin production in two chemotypes of Fusarium graminearum isolates. The composition of the oil phase, i.e., ripening inhibitor type and concentration, in clove oil-in-water nanoemulsions had a remarkable impact on antifungal activity as well as inhibition of mycotoxin production. In general, under the same clove oil concentration in oil phase, the addition of MCT decreased the antifungal and mycotoxin inhibitory activity of clove oil more than corn oil. Compared with the bulk clove oil, this study also indicated that the mycotoxin inhibitory activity of clove was significantly enhanced when encapsulated in nanoemulsions. These results have important implications for the design of essential oil based nanoemulsions as effective antifungal and detoxification delivery systems in the food or other industries.

Sfp-type 4'-phosphopantetheinyl transferase is required for lysine synthesis, tolerance to oxidative stress and virulence in the plant pathogenic fungus Cochliobolus sativus.

Polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs) are the major enzymes involved in the biosynthesis of secondary metabolites, which have diverse activities, including roles as pathogenicity/virulence factors in plant pathogenic fungi. These enzymes are activated by 4'-phosphopantetheinylation at the conserved serine residues, which is catalysed by 4'-phosphopantetheinyl transferase (PPTase). PPTase is also required for primary metabolism (α-aminoadipate reductase, AAR). In the genome sequence of the cereal fungal pathogen Cochliobolus sativus, we identified a gene (PPT1) orthologous to the PPTase-encoding genes found in other filamentous ascomycetes. The deletion of PPT1 in C. sativus generated mutants (Δppt1) that were auxotrophic for lysine, unable to synthesize melanin, hypersensitive to oxidative stress and significantly reduced in virulence to barley cv. Bowman. To analyse the pleiotropic effects of PPT1, we also characterized deletion mutants for PKS1 (involved in melanin synthesis), AAR1 (for AAR) and NPS6 (involved in siderophore-mediated iron metabolism). The melanin-deficient strain (Δpks1) showed no differences in pathogenicity and virulence compared with the wild-type strain. Lysine-auxotrophic mutants (Δaar1) induced spot blotch symptoms, as produced by the wild-type strain, when inoculated on wounded barley leaves or when lysine was supplemented. The Δnps6 strain showed a slightly reduced virulence compared with the wild-type strain, but exhibited significantly higher virulence than the Δppt1 strain. Our results suggest that an unknown virulence factor, presumably synthesized by PKSs or NRPSs which are activated by PPTase, is directly responsible for high virulence of C. sativus on barley cv. Bowman.