Effect of liquid-state fermentation on the antioxidant and functional properties of raw and roasted buckwheat flours.

The influence of liquid-state fermentation (LSF) by selected lactic acid bacteria (LAB) and Rhizopus oligosporus fungi on the content of rutin and total phenolic compounds (TPC), antioxidant capacity measured by ABTS test, FRAP assay and photochemiluminescence technique, and the inhibitory activity against formation of fluorescent advanced glycation end-products (AGEs) in vitro of raw and roasted buckwheat flours was studied. LSF caused a slight, specific LAB-dependent increase in TPC and a decrease in rutin content. Fermented raw buckwheat flours contained higher amounts of rutin and TPC with one exception when the highest increase in TPC was noted in roasted flour fermented by fungi. A LAB-dependent difference in the antioxidant capacity of buckwheat flours was noted while the inhibitory activity of fermented flours against AGEs formation was generally reduced. It can be concluded that LSF with selected LAB and fungi may improve the antioxidant and functional properties of buckwheat flours.

Effects of hot air treatment in combination with Pichia guilliermondii on postharvest preservation of peach fruit.

BACKGROUND: Antagonistic yeast and hot air treatment are two promising methods for conferring resistance to pathogenic fungi. The study assessed the effectiveness of hot-air treatment (45 °C, 4 h) and antagonistic yeast (Pichia guilliermondii at 108 CFU mL-1 ) alone or in combination on the two major postharvest diseases (Rhizopus stolonifer and Penicillium expansum), as well as the quality and antioxidant parameters in harvested peaches. RESULTS: The combination of hot-air treatment and Pichia guilliermondii had notable inhibitory effects on infections in peach fruit wounds. In addition, the individual hot-air treatment or Pichia guilliermondii could improve quality indexes to varying degrees, but the combination of the above two treatments could achieve the highest efficacy. Furthermore, compared with other groups, the combined treatment induced the highest activities of superoxide dismutase and catalase, improved the content of total phenolics and reduced glutathione most obviously. Lastly, the most significant reductions in malondialdehyde content and relative electrical conductivity were observed in the combination-treated fruit. CONCLUSIONS: The combined treatment could control fungal diseases, besides delay the decline of quality and antioxidant parameters, so as to achieve the purpose of fresh keeping for harvested peach fruit. © 2018 Society of Chemical Industry.

Improvement of the Culture Medium for the Dichlorvos-Ammonia (DV-AM) Method to Selectively Detect Aflatoxigenic Fungi in Soil.

The dichlorvos-ammonia (DV-AM) method is a simple but sensitive visual method for detecting aflatoxigenic fungi. Here we sought to develop a selective medium that is appropriate for the growth of aflatoxigenic fungi among soil mycoflora. We examined the effects of different concentrations of carbon sources (sucrose and glucose) and detergents (deoxycholate (DOC), Triton X-100, and Tween 80) on microorganisms in soils, using agar medium supplemented with chloramphenicol. The results demonstrated that 5⁻10% sucrose concentrations and 0.1⁻0.15% DOC concentrations were appropriate for the selective detection of aflatoxigenic fungi in soil. We also identified the optimal constituents of the medium on which the normal rapid growth of Rhizopus sp. was completely inhibited. By using the new medium along with the DV-AM method, we succeeded in the isolation of aflatoxigenic fungi from non-agricultural fields in Fukui city, Japan. The fungi were identified as Aspergillus nomius based on their calmodulin gene sequences. These results indicate that the new medium will be useful in practice for the detection of aflatoxigenic fungi in soil samples including those from non-agricultural environments.

Iron restriction inside macrophages regulates pulmonary host defense against Rhizopus species.

Mucormycosis is a life-threatening respiratory fungal infection predominantly caused by Rhizopus species. Mucormycosis has incompletely understood pathogenesis, particularly how abnormalities in iron metabolism compromise immune responses. Here we show how, as opposed to other filamentous fungi, Rhizopus spp. establish intracellular persistence inside alveolar macrophages (AMs). Mechanistically, lack of intracellular swelling of Rhizopus conidia results in surface retention of melanin, which induces phagosome maturation arrest through inhibition of LC3-associated phagocytosis. Intracellular inhibition of Rhizopus is an important effector mechanism, as infection of immunocompetent mice with swollen conidia, which evade phagocytosis, results in acute lethality. Concordantly, AM depletion markedly increases susceptibility to mucormycosis. Host and pathogen transcriptomics, iron supplementation studies, and genetic manipulation of iron assimilation of fungal pathways demonstrate that iron restriction inside macrophages regulates immunity against Rhizopus. Our findings shed light on the pathogenetic mechanisms of mucormycosis and reveal the role of macrophage-mediated nutritional immunity against filamentous fungi.

Effect on tomato plant and fruit of the application of biopolymer-oregano essential oil coatings.

BACKGROUND: Oregano essential oil (EO) was incorporated into film-forming dispersions (FFDs) based on biopolymers (chitosan and/or methylcellulose) at two different concentrations. The effect of the application of the FFDs was evaluated on tomato plants (cultivar Micro-Tom) at three different stages of development, and on pre-harvest and postharvest applications on tomato fruit. RESULTS: The application of the FFDs at '3 Leaves' stage caused phytotoxic problems, which were lethal when the EO was applied without biopolymers. Even though plant growth and development were delayed, the total biomass and the crop yield were not affected by biopolymer-EO treatments. When the FFDs were applied in the 'Fruit' stage the pre-harvest application of FFDs had no negative effects. All FFDs containing EO significantly reduced the respiration rate of tomato fruit and diminished weight loss during storage. Moreover, biopolymer-EO FFDs led to a decrease in the fungal decay of tomato fruit inoculated with Rhizopus stolonifer spores, as compared with non-treated tomato fruit and those coated with FFDs without EO. CONCLUSION: The application of biopolymer-oregano essential oil coatings has been proven to be an effective treatment to control R. stolonifer in tomato fruit. © 2016 Society of Chemical Industry.

Modification of Prenylated Stilbenoids in Peanut (Arachis hypogaea) Seedlings by the Same Fungi That Elicited Them: The Fungus Strikes Back.

Aspergillus oryzae and Rhizopus oryzae were compared for inducing the production of prenylated stilbenoids in peanut seedlings. The fungus was applied at two different time points: directly after soaking (day 1) or after 2 days of germination (day 3). Aspergillus- and Rhizopus-elicited peanut seedlings accumulated an array of prenylated stilbenoids, with overlap in compounds induced, but also with compounds specific to the fungal treatment. The differences were confirmed to be due to modification of prenylated stilbenoids by the fungus itself. Each fungus appeared to deploy different strategies for modification. The content of prenylated stilbenoids modified by fungi accounted for around 8% to 49% (w/w) of total stilbenoids. The contents of modified prenylated stilbenoids were higher when the fungus was applied on day 1 instead of day 3. Altogether, type of fungus and time point of inoculation appeared to be crucial parameters for optimizing accumulation of prenylated stilbenoids in peanut seedlings.

Preparation and application of a novel bioflocculant by two strains of Rhizopus sp. using potato starch wastewater as nutrilite.

A complex bioflocculant MBF917 was prepared by Rhizopus sp. M9 and M17 using potato starch wastewater (PSW) as nutrilite, and its flocculation characteristics of treating PSW were studied. Culture conditions of the two strains were optimized, and flocculating conditions of the bioflocculant for treating PSW were also investigated. The optimal and economical culture conditions were determined as COD of about 1600 mg/L, 0.3 g/L urea and 0.04 g/L potassium dihydrogen phosphate, with no need of adding carbon sources or adjusting pH. When the bioflocculant was used to flocculate PSW, the optimal dosage was 0.1 mL/L with addition of 5 mL/L 10% CaCl2 as coagulant aid, and there was no need to adjust pH. After flocculation, COD and turbidity removal rates of the PSW could reach 54.09% and 92.11%, respectively, and 1.1g/L proteic substance was recycled from the PSW as a byproduct that could be used for animal feed.

Induction of Rhizopus oryzae germination under starvation using host metabolites increases spore susceptibility to heat stress.

Sweetpotato is a nutritional source worldwide. Soft rot caused by Rhizopus spp. is a major limiting factor in the storage of produce, rendering it potentially unsafe for human consumption. In this study, Rhizopus oryzae was used to develop a concept of postharvest disease control by weakening the pathogen through induction of spore germination under starvation conditions. We isolated the sweetpotato active fractions (SPAFs) that induce spore germination and used them at a low dose to enhance spore weakening caused by starvation. Germination in SPAF at 1 mg/ml weakened the pathogen spores by delaying their ability to form colonies on rich media and by increasing their sensitivity to heat stress. The weakening effect was also supported by reduced metabolic activity, as detected by Alarmar Blue fluorescent dye assays. Spores incubated with SPAF at 1 mg/ml showed DNA fragmentation in some of their nuclei, as observed by TUNEL assay. In addition, these spores exhibited changes in ultrastructural morphology (i.e., shrinkage of germ tubes, nucleus deformation, and vacuole formation) which are hallmarks of programmed cell death. We suggest that induction of spore germination under starvation conditions increases their susceptibility to stress and, therefore, might be considered a new strategy for pathogen control.

Phytase production by Rhizopus microsporus var. microsporus biofilm: characterization of enzymatic activity after spray drying in presence of carbohydrates and nonconventional adjuvants.

Microbial phytases are enzymes with biotechnological interest for the feed industry. In this article, the effect of spray-drying conditions on the stability and activity of extracellular phytase produced by R. microsporus var. microsporus biofilm is described. The phytase was spray-dried in the presence of starch, corn meal (>150 µm), soy bean meal (SB), corn meal (<150 µm) (CM), and maltodextrin as drying adjuvants. The residual enzyme activity after drying ranged from 10.7% to 60.4%, with SB and CM standing out as stabilizing agents. Water concentration and residual enzyme activity were determined in obtained powders as a function of the drying condition. When exposed to different pH values, the SB and CM products were stable, with residual activity above 50% in the pH range from 4.5 to 8.5 for 60 min. The use of CM as drying adjuvant promoted the best retention of enzymatic activity compared with SB. Spray drying of the R. microsporus var. microsporus phytase using different drying adjuvants showed interesting results, being quite feasible with regards their biotechnological applications, especially for poultry diets.

Glyceollins, one of the phytoalexins derived from soybeans under fungal stress, enhance insulin sensitivity and exert insulinotropic actions.

Glyceollins are a category of phytoalexins that are produced by soybeans under fungal stress, but their effects on glucose homeostasis remain unknown. We hypothesized that glyceollins play an important role in glucose homeostasis by regulating glucose utilization in adipocytes and improving beta-cell function and survival. Glyceollins improved insulin-stimulated glucose uptake in 3T3-L1 adipocytes without activating the peroxisome proliferator-activated receptor-gamma agonist. They decreased triacylglycerol accumulation in adipocytes. In addition, glyceollins slightly improved glucose-stimulated insulin secretion without palmitate treatment in Min6 cells, and they potentiated insulinotropic actions when 500 microM palmitate was used to induce beta-cell dysfunction. This was associated with decreased beta-cell apoptosis because of the attenuation of endoplasmic reticulum stress, as determined by mRNA levels of XBP-1, ATF-4, ATF-6, and CHOP. Glyceollins also potentiated GLP-1 secretion to enhance insulinotropic actions in enteroendocrine cells. In conclusion, glyceollins help normalize glucose homeostasis by potentiating beta-cell function and survival and improving glucose utilization in adipocytes.

Effect of spores of saprophytic fungi on phytoalexin accumulation in seeds of frog-eye leaf spot and stem canker-resistant and -susceptible soybean (Glycine max L.) cultivars.

Two saprophytic fungi (Mucor ramosissimus and Rhizopus sp.) were tested for their ability to induce phytoalexin production by seeds of frog-eye leaf spot and stem canker-resistant and -susceptible soybean (Glycine max L.) cultivars. Only M. ramosissimus was shown to elicit a response and qualitative differences in phytoalexin accumulation were found between the susceptible and resistant cultivars. Glyceollins I, II, and III and glycinol were isolated from the susceptible cultivar, whereas Glyceollins I, II, and III, glycinol, glyceocarpin, genistein, isoformononetin, and N-acetyltyramine accumulated in the resistant cultivar in response to the same fungal elicitor. Genistein was found to be an inducibly formed isoflavonoid instead of a constitutive metabolite in the resistant cultivar, whereas N-acetyltyramine is described for the first time as a soybean phytoalexin. All the compounds, except genistein, showed fungitoxic activity against Cladosporium sphaerospermum. Spectral data of the pterocarpan phytoalexins, genistein, and N-acetyltyramine are also given in this work.

Evaluation of substrates and storage conditions for preparing and maintaining starter cultures for tempeh fermentation.

Heat-pasteurized cassava root, cowpeas, partially defatted peanuts, rice and soybeans were evaluated for their suitability to support growth and sporulation of the tempeh mold, Rhizopus microsporus var. oligosporus, and the oncom mold, Neurospora intermedia, at 25, 30 and 37 degrees C. The molds grew best and sporulated most luxuriantly on cassava and rice incubated at 37 degrees C. Viability of molds remained high for up to 30 weeks when dried (a(w) 0.48), powdered substrates on which the molds had been cultured, were stored at 5, 25 and 37 degrees C. Survival was best when powders were stored at 5 degrees C, although storage at 25 degrees C did not cause a marked decline in CFU/g. R. microsporus var. oligosporus starter cultures produced on rice and then stored for 20 weeks at these temperatures compared most favorably with commercial starter cultures for preparing high-quality soybean tempeh. The simple technology required to prepare these starter cultures enhances the potential for their application in developing countries where the introduction of nontraditional fermented legume foods to low-protein diets could help to ameliorate malnutrition.