Riboflavin induces Metarhizium spp. to produce conidia with elevated tolerance to UV-B, and upregulates photolyases, laccases and polyketide synthases genes.

AIMS: The effect of nutritional supplementation of two Metarhizium species with riboflavin (Rb) during production of conidia was evaluated on (i) conidial tolerance (based on germination) to UV-B radiation and on (ii) conidial expression following UV-B irradiation, of enzymes known to be active in photoreactivation, viz., photolyase (Phr), laccase (Lac) and polyketide synthase (Pks). METHODS AND RESULTS: Metarhizium acridum (ARSEF 324) and Metarhizium robertsii (ARSEF 2575) were grown either on (i) potato dextrose agar medium (PDA), (ii) PDA supplemented with 1% yeast extract (PDAY), (iii) PDA supplemented with Rb (PDA+Rb), or (iv) PDAY supplemented with Rb (PDAY+Rb). Resulting conidia were exposed to 866·7 mW m-2 of UV-B Quaite-weighted irradiance to total doses of 3·9 or 6·24 kJ m-2 . Some conidia also were exposed to 16 klux of white light (WL) after being irradiated, or not, with UV-B to investigate the role of possible photoreactivation. Relative germination of conidia produced on PDA+Rb (regardless Rb concentration) or on PDAY and exposed to UV-B was higher compared to conidia cultivated on PDA without Rb supplement, or to conidia suspended in Rb solution immediately prior to UV-B exposure. The expression of MaLac3 and MaPks2 for M. acridum, as well as MrPhr2, MrLac1, MrLac2 and MrLac3 for M. robertsii was higher when the isolates were cultivated on PDA+Rb and exposed to UV-B followed by exposure to WL, or exposed to WL only. CONCLUSIONS: Rb in culture medium increases the UV-B tolerance of M. robertsii and M. acridum conidia, and which may be related to increased expression of Phr, Lac and Pks genes in these conidia. SIGNIFICANCE AND IMPACT OF THE STUDY: The enhanced UV-B tolerance of Metarhizium spp. conidia produced on Rb-enriched media may improve the effectiveness of these fungi in biological control programs.

Clinical laser treatment of toenail onychomycoses.

Onychomycoses are fungal infections of the fingernails or toenails having a prevalence of 3% among adults and accounts for 50% of nail infections. It is caused by dermatophytes, non-dermatophyte filamentous fungi, and yeasts. Compressions and microtraumas significantly contribute to onychomycosis. Laser and photodynamic therapies are being proposed to treat onychomycosis. Laser light (1064 nm) was used to treat onychomycosis in 156 affected toenails. Patients were clinically followed up for 9 months after treatment. Microbiological detection of fungal presence in lesions was accomplished. A total of 116 samples allowed the isolation of at least a fungus. Most of nails were affected in more than two thirds surface (some of them in the full surface). In 85% of cases, after 18 months of the onset of treatment, culture turned negative. After 3 months months, only five patients were completely symptom-free with negative culture. In 25 patients, only after 6 months, the absence of symptoms was achieved and the cultures negativized; in 29 patients, 9 months were required. No noticeable adverse effects were reported. This study reinforces previous works suggesting the applicability of laser therapies to treat toenail onychomycosis.

Impact of infrared treatment on quality and fungal decontamination of mung bean (Vigna radiata L.) inoculated with Aspergillus spp.

BACKGROUND: Mung bean is a rich source of protein, carbohydrates and fiber content. It also exhibits a high level of antioxidant activity due to the presence of phenolic compounds. Aspergillus flavus and A. niger are the two major fungal strains associated with stored mung bean that lead to post-harvest losses of grains and also cause serious health risks to human beings. Thus there is a need to explore an economical decontamination method that can be used without affecting the biochemical parameters of grains. RESULTS: It was observed that infrared (IR) treatment of mung bean surface up to 70 °C for 5 min at an intensity of 0.299 kW m-2 led to complete visible inhibition of fungal growth. Scanning electron microscopy revealed that surface irregularities and physical disruption of spores coat are the major reasons behind the inactivation of IR-treated fungal spores. It was also reported that IR treatment up to 70 °C for 5 min does not cause any negative impact on the biochemical and physical properties of mung bean. CONCLUSION: From the results of the present study, it was concluded that IR treatment at 70 °C for 5 min using an IR source having an intensity of 0.299 kW m-2 can be successfully used as a method of fungal decontamination. The fungal spore population was reduced (approximately 5.3 log10 CFU g-1 reductions) without significantly altering the biochemical and physical properties of grains. © 2017 Society of Chemical Industry.

A novel photosensitization treatment for the inactivation of fungal spores and cells mediated by curcumin.

The global concerns regarding the emergence of fungicide-resistant strains and the impact of the excessive use of fungicidal practises on our health, food, and environment have increased, leading to a demand for alternative clean green technologies as treatments. Photosensitization is a treatment that utilises a photosensitiser, light and oxygen to cause cell damage to microorganisms. The effect of photosensitization mediated by curcumin on Aspergillus niger, Aspergillus flavus, Penicillium griseofulvum, Penicillium chrysogenum, Fusarium oxysporum, Candida albicans and Zygosaccharomyces bailii was investigated using three methods. The viability of spores/cells suspended in aqueous buffer using different concentrations of curcumin solution (100-1000µM) and light dose (0, 24, 48, 72 and 96J/cm2) were determined. Spraying curcumin solution on inoculated surfaces of agar plates followed by irradiation and soaking spores/cells in curcumin solution prior to irradiation was also investigated. In aqueous mixtures, photosensitised spores/cells of F. oxysporum and C. albicans were inhibited at all light doses and curcumin concentrations, while inactivation of A. niger, A. flavus P. griseofulvum, P. chrysogenum and Z. bailii were highly significant (P<0.001) reduced by 99%, 88.9%, 78%, 99.7% and 99.2% respectively. On the surface of agar plates, spores/cells exposed to a light dose of 360J/cm2 sprayed with curcumin at 800µM showed complete inhibition for A. niger, F. oxysporum, C. albicans and Z. bailii, while A. flavus P. griseofulvum, and P. chrysogenum reduced by 75%, 80.4% and 88.5% respectively. Soaking spores/cells with curcumin solution prior to irradiation did not have a significant effect on the percentage reduction. These observations suggest that a novel photosensitization mediated curcumin treatment is effective against fungal spores/cells and the variation of percentage reduction was dependent on curcumin concentration, light dosage and fungal species.

Rhizophagus irregularis MUCL 41833 can colonize and improve P uptake of Plantago lanceolata after exposure to ionizing gamma radiation in root organ culture.

Long-lived radionuclides such as (90)Sr and (137)Cs can be naturally or accidentally deposited in the upper soil layers where they emit ß/γ radiation. Previous studies have shown that arbuscular mycorrhizal fungi (AMF) can accumulate and transfer radionuclides from soil to plant, but there have been no studies on the direct impact of ionizing radiation on AMF. In this study, root organ cultures of the AMF Rhizophagus irregularis MUCL 41833 were exposed to 15.37, 30.35, and 113.03 Gy gamma radiation from a (137)Cs source. Exposed spores were subsequently inoculated to Plantago lanceolata seedlings in pots, and root colonization and P uptake evaluated. P. lanceolata seedlings inoculated with non-irradiated AMF spores or with spores irradiated with up to 30.35 Gy gamma radiation had similar levels of root colonization. Spores irradiated with 113.03 Gy gamma radiation failed to colonize P. lanceolata roots. P content of plants inoculated with non-irradiated spores or of plants inoculated with spores irradiated with up to 30.35 Gy gamma radiation was higher than in non-mycorrhizal plants or plants inoculated with spores irradiated with 113.03 Gy gamma radiation. These results demonstrate that spores of R. irregularis MUCL 41833 are tolerant to chronic ionizing radiation at high doses.

Comparative Sensitivity of Trichophyton and Aspergillus Conidia to Inactivation by Violet-Blue Light Exposure.

OBJECTIVE: To investigate the use of 405 nm light for inhibiting the growth of selected species of dermatophytic and saprophytic fungi. BACKGROUND DATA: The increasing incidence and resilience of dermatophytic fungal infections is a major issue, and alternative treatment methods are being sought. METHODS: The sensitivity of the dermatophytic fungi Trichophyton rubrum and Trichophyton mentagrophytes to 405 nm violet-blue light exposure was investigated, and the results compared with those obtained with the saprophytic fungus Aspergillus niger. Microconidia of T. rubrum and T. mentagrophytes and conidia of A. niger were seeded onto Sabauroud dextrose agar plates and irradiated with 405 nm light from an indium-gallium-nitride 99-DIE light-emitting diode (LED) array and the extent of inhibition was measured. RESULTS: Germination of the microconidia of the Trichophyton species was completely inhibited using an irradiance of 35 mW/cm(2) for 4 h (dose of 504 J/cm(2)). A. niger conidia showed greater resistance, and colonial growth developed after light exposure. In liquid suspension tests, 405 nm light dose levels of 360, 720, and 1440 J/cm(2) resulted in complete inactivation of T. rubrum microconidia, whereas A. niger showed greater resistance, and at the highest dose level applied (1440 J/cm(2)) although A niger hyphae were completely inactivated, only a 3-log10 reduction of a 5-log10 conidial suspension was achieved. CONCLUSIONS: The study results demonstrate the relatively high sensitivity of Trichophyton microconidia to 405 nm violet-blue light, and this is may be of potential interest regarding the control and treatment of dermatophyte infections.

Dark Period Following UV-C Treatment Enhances Killing of Botrytis cinerea Conidia and Controls Gray Mold of Strawberries.

Strawberries are available throughout the year either from production in the field or from high and low tunnel culture. Diversity of production conditions results in new challenges in controlling diseases before and after harvest. Fungicides have traditionally been used to control these diseases; however, their limitations necessitate a search for new approaches. We found that UV-C irradiation of Botrytis cinerea, a major pathogen of strawberry, can effectively kill this fungus if a dark period follows the treatment. The inclusion of a 4-h dark period resulted in almost complete kill of B. cinerea conidia on agar media at a dose of 12.36 J/m2. The UV-C dose did not cause a reduction in photosynthesis in strawberry leaves or discoloration of sepals, even after exposing plants repeatedly (twice a week) for 7 weeks. Although irradiation of dry conidia of B. cinerea with this dose resulted in some survival, the conidia were not infective and not able to cause decay even when inoculated onto a highly susceptible mature apple fruit. Irradiation of strawberry pollen at 12.36 J/m2 did not affect pollen germination, tube growth and length in vitro, or germination and tube growth in the style of hand-pollinated emasculated strawberry flowers. No negative effect of the UV-C treatment was observed on fruit yield and quality in high tunnel culture. In the fruit and flower petal inoculation tests, the UV-C treatment was highly effective in reducing fruit decay and petal infection. This UV-C treatment with an exposure time of 60 s may be useful in controlling gray mold in tunnel production of strawberries and may also have the potential for use in intensive field and indoor production of other fruits and vegetables providing that a 4-h dark period follows the irradiation.

Variable Efficacy of the Proteinaceous Antifungal YvgO in Select Fruit Juices and Teas as a Complement with UV Methods of Food Protection.

Heat-resistant fungal spores present a processing challenge for beverages and fruit juices, as thermal and UV strategies are often inadequate in reducing heat-resistant fungal burdens to acceptable levels. While effective against pathogenic or invasive bacteria, germicidal UV light treatments also fail to achieve an appreciable reduction of heat-resistant fungal spores. As an alternative, the efficacy of the antifungal protein YvgO was examined across a selection of fruit juices and teas, as well as solid model matrices. Compared with its efficacy in analogous liquid matrices, the apparent efficacy of YvgO was diminished on acidified solid matrices due to a reduction in YvgO diffusion. Using an XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] tetrazolium dye cytotoxicity assay, the effective concentrations to reduce growth by 50% were elucidated in samples challenged with Byssochlamys fulva H25. The MICs were determined and ranged from 2 ppm in apple juice and acidified teas to approximately 3 to 12 ppm for lemonade and orange, white cranberry, blueberry, prune, cherry, and grape juices. Apple cider and nonacidified teas showed reduced efficacy, with MICs exceeding 100 ppm. Tannin-rich products readily removed YvgO from the product, impairing its efficacy. Adding bovine serum albumin as a competitive inhibitor effectively reversed the YvgO-tannin association and restored efficacy in black but not green tea matrices. When challenged with a 5-log CFU inoculum of B. fulva, the shelf lives of the products were extended for various times up to 28 days in a concentrationdependent manner. However, initial efficacy was not predictive of shelf life extension, as some products exhibited improved protection at just two- and fourfold concentrations above the MIC, while others only exhibited long-term stability when concentrations exceeded 20 times the MIC. As such, YvgO may be an attractive alternative to currently available protection strategies and will provide needed diversity for natural food protectants.

Cochliobolus lunatus colonizes potato by adopting different invasion strategies on cultivars: New insights on temperature dependent-virulence.

Extreme temperature fluctuations affect the interaction dynamics of Cochliobolus lunatus through temperature-dependent virulence, virulence differentiation and induced-virulence which poses a major threat to global food security. The relationship between higher temperature and pathogenicity of C. lunatus on reported hosts are poorly understood. In this study, temperature stress was applied on C. lunatus to investigate the correlation among the different types of conidia. Additionally, a comparative dissection of the invasion process, infection structures and conidial germination pattern on four different Solanum tuberosum L. (potato) cultivars were performed. Based on microscopic examination, it was found that C. lunatus adopts different hyphae morphology and septation pattern at different temperature regimes and produce different types of conidia. The study showed that four-celled conidia are overproduced at elevated temperature (>30 °C) than one, two, three and five-celled conidia. Our finding revealed that C. lunatus conidia exhibit bipolar germination (>14.67%, P<0.05), unipolar germination (>35.33%, P<0.05), penetrate subcutaneously via epidermal anticlinal cell wall (>0.33%, P < 0.05) and differentially form appressoria-like structures during colonization of four different potato cultivars. Importantly, it is shown that unipolar germination and bipolar germination in C. lunatus are independently occurring phenomenon irrespective of the host. It is confirmed that C. lunatus adopt different but highly successful strategies on four different potato cultivars to incite brown-to-black leaf spot disease. Altogether, our data showed that increase in temperature enhances C. lunatus virulence on different potato cultivars irrespective of their inherent thermotolerant traits.

Enhanced conidial thermotolerance and field efficacy of Beauveria bassiana by saccharides.

Insecticide efficacy of Beauveria bassiana conidia was improved by optimizing the concentrations of conidial heat-protective saccharides (glucose, sucrose, maltose, trehalose, α-lactose, and mannitol) using response surface methodology. Two field trials in tea gardens were carried out to control leafhopper (Empoasca vitis) by spraying B. bassiana conidia together with the optimized saccharides (0.26 g glucose, 0.28 g lactose, 0.24 g mannitol per ml). In the field studies, B. bassiana conidia were applied to control Empoasca vitis with and without saccharides and compared with bifenthrin, a pyrethroid insecticide. With the optimal concentrations of saccharides, the conidial germination rate reached 72% and the control efficacy of the saccharides group (65.7%) was equal to the bifenthrin group (69.4%), which improved by about 55%.

An investigation into the inhibitory effect of ultraviolet radiation on Trichophyton rubrum.

Fungal infection of nails, onychomycosis, is predominantly caused by Trichophyton rubrum. This infection is an important public health concern due to its persistent nature and high recurrence rates. Alternative treatments are urgently required. One such alternative is phototherapy involving the action of photothermal or photochemical processes. The aim of this novel study was to assess which wavelengths within the ultraviolet (UV) spectrum were inhibitory and equally important nail transmissible. Initial irradiations of T. rubrum spore suspensions were carried out using a tunable wavelength lamp system (fluence ≤3.1 J/cm(2)) at wavelengths between 280 and 400 nm (UVC to UVA) to evaluate which wavelengths prevented fungal growth. Light-emitting diodes (LEDs) of defined wavelengths were subsequently chosen with a view to evaluate and potentially implement this technology as a low-cost "in-home" treatment. Our experiments demonstrated that exposure at 280 nm using an LED with a fluence as low as 0.5 J/cm(2) was inhibitory, i.e., no growth following a 2-week incubation (p < 0.05; one-way ANOVA), while exposure to longer wavelengths was not. A key requirement for the use of phototherapy in the treatment of onychomycosis is that it must be nail transmissible. Our results indicate that the treatment with UVC is not feasible given that there is no overlap between the antifungal activity observed at 280 nm and transmission through the nail plate. However, a potential indirect application of this technology could be the decontamination of reservoirs of infection such as the shoes of infected individuals, thus preventing reinfection.

Spore Loads May Not be Used Alone as a Direct Indicator of the Severity of Nosema ceranae Infection in Honey Bees Apis mellifera (Hymenoptera:Apidae).

Nosema ceranae Fries et al., 1996, a microsporidian parasite recently transferred from Asian honey bees Apis cerana F., 1793, to European honey bees Apis mellifera L., 1758, has been suspected as one of the major culprits of the worldwide honey bee colony losses. Spore load is a commonly used criterion to describe the intensity of Nosema infection. In this study, by providing Nosema-infected bees with sterilized pollen, we confirmed that pollen feeding increased the spore loads of honey bees by several times either in the presence or absence of a queen. By changing the amount of pollen consumed by bees in cages, we showed that spore loads increased with an increase in pollen consumption. Nosema infections decrease honey bee longevity and transcription of vitellogenin, either with or without pollen feeding. However, the reduction of pollen consumption had a greater impact on honey bee longevity and vitellogenin level than the increase of spore counts caused by pollen feeding. These results indicate that spore loads may not be used alone as a direct indicator of the severity of N. ceranae infection in honey bees.

Inactivation of A. ochraceus spores and detoxification of ochratoxin A in coffee beans by gamma irradiation.

UNLABELLED: Ochratoxin A (OTA) produced in food by Aspergillus ochraceus is known to cause adverse health effects. Among the plantation products, green coffee beans are prone to fungal attack and get contaminated with OTA frequently. A fungal strain isolated from green coffee beans was characterized by morphological analyses as well as internal transcribed spacer (ITS) and 5.8S rDNA sequencing, turned out to be A. ochraceus, however, nontoxigenic. Hence, additional strains of A. ochraceus were procured and characterized for toxin production. Presterilized green coffee beans were spiked with a toxigenic strain and treated with gamma radiation. Minimum inhibitory dose (MID) of gamma radiation for 10(4) and 10(8) spores of A. ochraceus strain per 10 g of green coffee beans was found to be approximately 1 and approximately 2.5 kGy, respectively. The radiation treatment (10 kGy) almost degraded the preformed or in vitro added OTA (50 ppb) in coffee beans. OTA degradation was found to be enhanced with increase in moisture content. Cytotoxicity in terms of cell viability was found to be reduced significantly for radiation treated OTA in MTT [3-(4,5-dimethylthiazole-2yl)-2,5-diphenyl tetrazolium bromide] assay as well as flow cytometric analysis when studied using human intestinal epithelial (Int-407) cells. Similar finding was also observed with E. coli MG1655 cells. Thus the inclusion of gamma radiation treatment in the postharvest processing chain of green coffee beans could help in eliminating toxigenic fungi as well as destroying preformed OTA without affecting the sensory attributes. PRACTICAL APPLICATION: In general, mycotoxins including ochratoxin A (OTA) are highly stable to detoxifying agents. Green coffee beans are prone to fungal attack and could get frequently contaminated with the OTA due to improper drying or rehydration during storage. Gamma radiation processing of green coffee beans was found to eliminate the A. ochraceus spores as well as inactivate OTA without affecting its sensory attributes. Thus inclusion of gamma radiation in the postharvest processing chain of green coffee beans would be very useful for consumer safety and coffee trade.

Production of thermotolerant entomopathogenic Isaria fumosorosea SFP-198 conidia in corn-corn oil mixture.

Low thermotolerance of entomopathogenic fungi is a major impediment to long-term storage and effective application of these biopesticides under seasonal high temperatures. The effects of high temperatures on the viability of an entomopathogenic fungus, Isaria fumosorosea SFP-198 (KCTC 0499BP), produced on different substrates amended with various additives were explored. Ground corn was found to be superior in producing the most thermotolerant conidia compared to yellow soybean, red kidney bean, and rice in a polyethylene bag production system. Using ground corn mixed with corn oil as a substrate resulted in only 7% reduction in germination compared to ground corn alone (67% reduction) after exposure of conidia to 50 degrees C for 2 h. Corn oil as an additive for ground corn was followed by inorganic salts (KCl and NaCl), carbohydrates (sucrose and dextrin), a sugar alcohol (sorbitol), and plant oils (soybean oil and cotton seed oil) in ability to improve conidial thermotolerance. Unsaturated fatty acids, such as linoleic acid and oleic acid, the main components of corn oil, served as effective additives for conidial thermotolerance in a dosage-dependent manner, possibly explaining the improvement by corn oil. This finding suggests that the corn-corn oil mixture can be used to produce highly thermotolerant SFP-198 conidia and provides the relation of unsaturated fatty acids as substrates with conidial thermotolerance.

Use of UV-C treatment to inhibit the microbial growth and maintain the quality of Yali pear.

UNLABELLED: The effects of UV-C radiation on microbial growth in vitro (Monilinia fruticola) and in inoculated Yali pears (Pyrus bretschneideri Rehd.) were investigated. Moreover, postharvest quality and the activities of defense and antioxidant enzymes were analyzed after the pears were exposed to UV-C irradiation at an energy level of 5 kJ m⁻².The results showed that spore germination of M. fructicola was significantly inhibited by each of the 3 doses (1, 5, and 10 kJ m⁻²) in vitro. In the in vivo assays, lesion diameter on the fruit being inoculated before or after the UV-C treatment was both significantly lower than that on the fruit of control. Meanwhile, the activities of phenylalanine ammonia lyase, ß-1,3-glucanase, superoxide dismutase, catalase, and glutathione reductase were induced to high levels by UV-C treatment. We conclude that UV-C treatment could reduce postharvest disease by the germicidal and induced effects and maintain the quality by enhancing the antioxidant enzyme activities. PRACTICAL APPLICATION: UV-C radiation has recently been proposed as a new technology to avoid chemical fungicides. However, there are few studies regarding the effect of UV-C treatment on Yali pear. In this study, we found that 5 kJ m⁻² UV-C irradiation can control postharvest disease and maintain the quality of Yali pear. This method may be applied to reduce the decay of Yali pears during exporting and storage.

Cultural methods and environmental conditions affecting gray mold and its management in lisianthus.

Gray mold, caused by Botrytis cinerea, severely affects the base of the stems of lisianthus (Eustoma grandiflorum) plants as well as the cut stems left after flowers are harvested. This study examined infection of lisianthus plants by B. cinerea under laboratory and commercial greenhouse production conditions typical for Israel and evaluated cultural methods for manipulating disease development in commercial greenhouses. Although the lower nodes of lisianthus stems are typically infected, in this study, the inherent susceptibility of these nodes was less than that of nodes midway up the stem. Greater light intensity (4,860 lux) was associated with significantly more severe stem wounds than lower light intensities of 140 to 1,020 lux. Lower light intensity (140 lux) was associated with significantly more severe leaf infection. The development of gray mold along leaves toward the stem was slower at 26 degrees C than at 18 to 20 degrees C and was fastest at relative humidity (RH) levels close to saturation (>99%). B. cinerea infection developed in all stem wounds exposed to 65 to 99% RH and at temperatures of 12 to 29 degrees C. Infection severity in stem wounds (measured as lesion length) on whole plants was significantly less at 26 degrees C than at 18 or 22 degrees C, and was significantly higher at 99% RH compared with 70 to 85 and 85 to 95% RH. Severity of gray mold was the greatest at 15 to 22 degrees C and 85 to 99% RH. Under commercial greenhouse conditions, supplemental calcium (Ca(NO3)2) applied in fertigation or as a spray led to moderate yet significant reduction in disease severity. In addition, polyethylene soil cover and the use of buried drip irrigation instead of surface drip irrigation suppressed gray mold significantly on cut stems following harvest. Covering the soil with polyethylene also suppressed gray mold significantly as compared with the common practice of growing lisianthus in bare soil.

Optimization of production of monacolin K from gamma-irradiated Monascus mutant by use of response surface methodology.

Monascus isolate number 711, which is capable of producing monacolin K as an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, the key enzyme of cholesterol synthesis, was isolated from Ang-kak, the red yeast rice koji. To increase the monacolin K-producing activity of the strain, spore suspensions of the strain were subjected to gamma-irradiation. One thousand mutants were generated via gamma-irradiation and screened using bioassay and high performance liquid chromatography analysis. Several mutants with higher productivities of monacolin K than that of the parent strain were primarily selected. Mutant KU609 was finally selected because of its characteristics of high monacolin K production and non-citrinin-producing activity under our test conditions. Response surface methodology was used to analyze the effect of culture medium on the production of monacolin K in mixed solid-state cultures. The optimal values of nutritional ingredients for the maximal production were soytone, glucose, MgSO4, and barley at concentrations of 0.5 g, 0.48 g, 0.053 g, and 9 g, respectively. The final monacolin K production of Monascus KU609 was increased almost 100-fold compared to that of the parent strain.

Inactivation of conidia of Botrytis cinerea and Monilinia fructigena using UV-C and heat treatment.

The effect of UV-C (lambda = 254 nm) and heat treatment was investigated on the inactivation of conidia of Botrytis cinerea and Monilinia fructigena, two major postharvest spoilage fungi of strawberries and cherries, respectively. Both fungi were grown at 21 degrees C in the dark and conidia were isolated after 1 week by washing the mycelium with a mild detergent solution. After filtration and resuspension in phosphate buffer to a titer of 10(5) to 10(6) cfu/ml, the conidia were subjected to different treatments. The applied UV-C doses varied from 0.01 to 1.50 J/cm2, and the conditions for the thermal treatment were 3, 5, 10, 15 and 20 min at temperatures ranging from 35 to 48 degrees C. Both techniques were applied individually and in combination. Spore inactivation increased with increasing intensity of single treatments. No surviving spores of B. cinerea were observed after 15 min at 45 degrees C or an UV-C treatment of 1.00 J/cm2. M. fructigena was more sensitive and a thermal treatment of 3 min at 45 degrees C or an UV-C treatment of 0.50 J/cm2 resulted in complete spore inactivation. Combination of both techniques reduced the required intensity of the treatment for inactivation of both fungi. The order of the applications had a significant effect on the degree of inactivation. The inactivation of B. cinerea conidia was greater when the heat treatment came first, and for M. fructigena, most inactivation was achieved when the heat treatment was preceded with an UV-C irradiation.

Riboflavin, overproduced during sporulation of Ashbya gossypii, protects its hyaline spores against ultraviolet light.

Riboflavin (vitamin B2), essential in tiny amounts as a precursor for oxidoreductase coenzymes, is a yellow pigment. Although it causes cytotoxicity via photoinduced damage of macromolecules, several microorganisms are striking overproducers. A question, unanswered for decades, is whether riboflavin overproducers can benefit from this property. Here, we report an ultraviolet (UV) protective effect of riboflavin. The spores of Ashbya gossypii, a riboflavin-overproducing fungus, are more sensitive to UV than those of Aspergillus nidulans. The addition of riboflavin to suspensions improves the UV resistance of both spore types. Interestingly, we show that regulation of sporulation and riboflavin overproduction in A. gossypii are linked. In batch culture, both were elevated when growth ceased. At constant growth rates, obtained in a chemostat culture, neither was elevated. Supplementation of cultures by cAMP, a known stress signal, negatively affected sporulation as well as riboflavin overproduction, establishing a second, independent argument for the linkage.

Evaluation of drug phototoxicity by photosensitization of Trichophyton mentagrophytes.

Long wave ultraviolet radiation to suspensions of T. mentagrophytes, in the presence of photosensitizing chemicals, resulted in death of the fungus as measured by loss in colony-forming ability. Photosensitization could be demonstrated, with appropriate exposures and concentrations, with chlorpromazine, promethazine, perphenazine, anthracene, sulphanilamide, trimethylpsoralen, tolbutamide, tetrachlorosalicylanilide, and tribromosalicylanilide. No phototoxicity was detected with demethylchlortetracycline and thiazides. The present system seems to be simple, reproducible, and sensitive as an in vitro screening test for the photosensitizing potential of chemicals.