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Sample records for aspergillus flavus growth

  1. Inhibitory effects of gossypol-related compounds on growth of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gossypolone demonstrated growth inhibitory activity against Aspergillus flavus isolate AF13. Growth inhibition was concentration dependent, with a 50% effective dose of 90 µg gossypolone per mL of medium (165 µM). Growth inhibition levels of up to 95% were achieved with gossypolone concentrations ...

  2. Aspergillus flavus Blast2GO gene ontology database: elevated growth temperature alters amino acid metabolism

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The availability of a representative gene ontology (GO) database is a prerequisite for a successful functional genomics study. Using online Blast2GO resources we constructed a GO database of Aspergillus flavus. Of the predicted total 13,485 A. flavus genes 8,987 were annotated with GO terms. The mea...

  3. Hybridization between Aspergillus flavus and Aspergillus parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To date the sexual stages or teleomorphs have been described for three aflatoxigenic species in Aspergillus section Flavi: Petromyces flavus, P. parasiticus and P. nomius. In this study we examined the possibility of interspecific matings between A. flavus and A. parasiticus. These species can b...

  4. Controlling Aspergillus flavus and Aspergillus parasiticus growth and aflatoxin production in poultry feed using carvacrol and trans-cinnamaldehyde.

    PubMed

    Yin, Hsin-Bai; Chen, Chi-Hung; Kollanoor-Johny, Anup; Darre, Michael J; Venkitanarayanan, Kumar

    2015-09-01

    Aflatoxins (AF) are toxic metabolites primarily produced by molds, Aspergillus flavus and Aspergillus parasiticus. Contamination of poultry feed with AF is a major concern to the poultry industry due to severe economic losses stemming from poor performance, reduced egg production, and diminished egg hatchability. This study investigated the inhibitory effect of 2 generally regarded as safe (GRAS), natural plant compounds, namely carvacrol (CR) and trans-cinnamaldehyde (TC), on A. flavus and A. parasiticus growth and AF production in potato dextrose broth (PDB) and in poultry feed. In broth culture, PDB supplemented with CR (0%, 0.02%, 0.04% and 0.08%) or TC (0%, 0.005%, 0.01% and 0.02%) was inoculated with A. flavus or A. parasiticus (6 log CFU/mL), and mold counts and AF production were determined on days 0, 1, 3, and 5. Similarly, 200 g portions of poultry feed supplemented with CR or TC (0%, 0.4%, 0.8%, and 1.0%) were inoculated with each mold, and their counts and AF concentrations in the feed were determined at 0, 1, 2, 3, 4, 8, and 12 weeks of storage. Moreover, the effect of CR and TC on the expression of AF synthesis genes in A. flavus and A. parasiticus (aflC, nor1, norA, and ver1) was determined using real-time quantitative PCR (RT-qPCR). All experiments had duplicate samples and were replicated 3 times. Results indicated that CR and TC reduced A. flavus and A. parasiticus growth and AF production in broth culture and chicken feed (P<0.05). All tested concentrations of CR and TC decreased AF production in broth culture and chicken feed by at least 60% when compared to controls (P<0.05). In addition, CR and TC down-regulated the expression of major genes associated with AF synthesis in the molds (P<0.05). Results suggest the potential use of CR and TC as feed additives to control AF contamination in poultry feed. PMID:26217023

  5. The effect of water activity and storage temperature on the growth of Aspergillus flavus in medicinal herbs.

    PubMed

    Kulshrestha, Rashmi; Gupta, Chandra Prakash; Shukla, Gyanesh; Kundu, Madan Gopal; Bhatnagar, Satyendra Prasad; Katiyar, Chandra Kant

    2008-08-01

    The quality control of medicinal herbs post harvesting or after collection becomes very critical because of susceptibility to fungal invasion during storage depending on the temperature and humidity of the storage area. The information on moisture equilibrium is important on the process and storage of foods which can be extended to medicinal herbs. In the present study, the growth of Aspergillus flavus was observed on selected ten medicinal herbs with water activity aw above 0.81 when stored at 25 +/- 2 degrees C, 30 +/- 2 degrees C and 40 +/- 2 degrees C except for Picrorhiza kurrooa and Alpinia galanga which were found to have anti-fungal properties. Aspergillus flavus did not grow in any samples of medicinal herbs with water activity aw below 0.81 at temperatures of 25 +/- 2 degrees C, 30 +/- 2 degrees C and 40 +/- 2 degrees C. Also Aspergillus flavus did not grow in any samples of medicinal herbs with water activity aw above 0.81 when stored below 10 +/- 2 degrees C. Therefore it can be concluded that the contamination of medicinal herbs with aflatoxins can be minimized by controlling water activity and storage temperature. Sorption isotherms (desorption) can be interpreted to determine the optimum drying which can lower the water activity to the level required for preventing growth of Aspergillus flavus and also for ensuring quality of medicinal herbs which may get destroyed upon over drying. Furthermore, it also saves incremental cost in prolonged drying over the optimum drying. PMID:18553273

  6. The Inhibitory Effects of Curcuma longa L. Essential Oil and Curcumin on Aspergillus flavus Link Growth and Morphology

    PubMed Central

    Mossini, Simone Aparecida Galerani; Ferreira, Francine Maery Dias; Arrotéia, Carla Cristina; da Costa, Christiane Luciana; Nakamura, Celso Vataru; Machinski Junior, Miguel

    2013-01-01

    The essential oil from Curcuma longa L. was analysed by GC/MS. The major components of the oil were ar-turmerone (33.2%), α-turmerone (23.5%) and β-turmerone (22.7%). The antifungal activities of the oil were studied with regard to Aspergillus flavus growth inhibition and altered morphology, as preliminary studies indicated that the essential oil from C. longa inhibited Aspergillus flavus Link aflatoxin production. The concentration of essential oil in the culture media ranged from 0.01% to 5.0% v/v, and the concentration of curcumin was 0.01–0.5% v/v. The effects on sporulation, spore viability, and fungal morphology were determined. The essential oil exhibited stronger antifungal activity than curcumin on A. flavus. The essential oil reduced the fungal growth in a concentration-dependent manner. A. flavus growth rate was reduced by C. longa essential oil at 0.10%, and this inhibition effect was more efficient in concentrations above 0.50%. Germination and sporulation were 100% inhibited in 0.5% oil. Scanning electron microscopy (SEM) of A. flavus exposed to oil showed damage to hyphae membranes and conidiophores. Because the fungus is a plant pathogen and aflatoxin producer, C. longa essential oil may be used in the management of host plants. PMID:24367241

  7. Volatile trans-2-hexenal a soybean aldehyde inhibits Aspergillus flavus growth and aflatoxin production in corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trans-2-hexenal, a volatile aldehyde, is produced by soybean [Glycine max (L.) Merr] and other plants via the lipoxygenase pathway. In vitro tests showed it significantly (p< 0.001) reduced Aspergillus flavus germinating conidial viability at 10 µM, with approximately 95% viability reduction observ...

  8. 76 FR 16297 - Aspergillus flavus

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-23

    ...This regulation establishes an exemption from the requirement of a tolerance for residues of the microbial pesticide, Aspergillus flavus AF36, in or on corn food and feed commodities, when applied/used as an antifungal agent. The Arizona Cotton Research and Protection Council submitted a petition to EPA under the Federal Food, Drug, and Cosmetic Act (FFDCA), requesting an amendment to the......

  9. Sexual reproduction in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is the major producer of carcinogenic aflatoxins in crops worldwide and is also an important opportunistic human pathogen in aspergillosis. The sexual state of this heterothallic fungus is described from crosses between strains of the opposite mating type. Sexual reproduction oc...

  10. Effect of temperature and water activity on growth and aflatoxin production by Aspergillus flavus and Aspergillus parasiticus on cured meat model systems.

    PubMed

    Peromingo, Belén; Rodríguez, Alicia; Bernáldez, Victoria; Delgado, Josué; Rodríguez, Mar

    2016-12-01

    Dry-cured hams may be colonised by aflatoxin-producing Aspergillus flavus and Aspergillus parasiticus during the ripening process. The objective of this study was to evaluate the interaction between non-ionic water stress and temperatures may have on lag phases prior to growth, growth rates and aflatoxin production by two strains of each A. parasiticus and A. flavus on meat matrices over a period of 12days. Results showed that A. flavus CBS 573.65 had shorter lag phases than A. parasiticus CECT 2688, however the growth rates were quite similar. For both species, no growth occurred at 10°C and all aw tested and optimum growth happened at 25°C and 0.95 aw. Similar aflatoxin B1 production profiles between both species were found, however A. flavus produced much higher concentration of such toxin than A. parasiticus. Both species produced aflatoxins when the temperature and the aw were ≥15°C and ≥0.90. PMID:27498402

  11. NITRIFICATION BY ASPERGILLUS FLAVUS1

    PubMed Central

    Marshall, K. C.; Alexander, M.

    1962-01-01

    Marshall, K. C. (Cornell University, Ithaca, N. Y.) and M. Alexander. Nitrification by Aspergillus flavus. J. Bacteriol. 83:572–578. 1962.—Aspergillus flavus has been shown to produce bound hydroxylamine, nitrite, and nitrate when grown in peptone, amino acid, or buffered ammonium media. Free hydroxylamine was not detected in these cultures, but it was found in an unbuffered ammonium medium in which neither nitrite nor nitrate was formed. Evidence was obtained for the presence of β-nitropropionic acid in the filtrate of an actively nitrifying culture. Alumina treatment of an ammonium medium prevented the formation by growing cultures of nitrite and nitrate but not bound hydroxylamine. The effect of alumina treatment was reversed by the addition of 10−3m CeCl3 to the medium. Extracts of the fungus contained peroxidase and an enzyme capable of catalyzing the production of nitrite from β-nitropropionic acid. The nitrite-forming enzyme is apparently specific for β-nitropropionate; no activity was found with nitromethane, nitroethane, and nitropropane as substrates. Nitrate was not reduced to nitrite nor was nitrite oxidized to nitrate by the hyphal extracts. The significance of these observations in nitrification by A. flavus is discussed. PMID:14470254

  12. Buckwheat achenes antioxidant profile modulates Aspergillus flavus growth and aflatoxin production.

    PubMed

    Chitarrini, G; Nobili, C; Pinzari, F; Antonini, A; De Rossi, P; Del Fiore, A; Procacci, S; Tolaini, V; Scala, V; Scarpari, M; Reverberi, M

    2014-10-17

    Buckwheat (Fagopyrum spp.) is a "pseudo-cereal" of great interest in the production of healthy foods since its flour, derived from achenes, is enriched with bioactive compounds and, due to the absence of gluten, may be used in composition of celiac diets. Amongst buckwheat species, F. tataricum achenes possess a larger amount of the antioxidant flavenol rutin than the common buckwheat F. esculentum. Ongoing climate change may favor plant susceptibility to the attack by pathogenic, often mycotoxigenic, fungi with consequent increase of mycotoxins in previously unexploited feeds and foodstuffs. In particular, Aspergillus flavus, under suitable environmental conditions such as those currently occurring in Italy, may produce aflatoxin B1 (AFB1), the most carcinogenic compound of fungal origin which is classified by IARC as Category 1. In this study, the viable achenes of two buckwheat species, F. tataricum (var. Golden) and F. esculentum (var. Aelita) were inoculated with an AFB1-producing A. flavus NRRL 3357 to analyze their relative performances against fungal invasion and toxin contamination. Notably, we sought the existence of a correlation between the amount of tocols/flavonols in the achenes of buckwheat, infected and non-infected with A. flavus, and to analyze the ability of the pathogen to grow and produce toxin during achene infection. Results suggest that achenes of F. tataricum, the best producer of antioxidant compounds in this study, are less susceptible to A. flavus infection and consequently, but not proportionally, to mycotoxin contamination compared with F. esculentum. Moreover, rutin-derived quercetin appears to be more efficient in inhibiting aflatoxin biosynthesis than the parent compound. PMID:25108759

  13. The role of Aspergillus flavus veA in the production of extracellular proteins during growth on starch substrates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aflatoxin-producer and opportunistic plant pathogenic, filamentous fungus Aspergillus flavus is responsible for the contamination of corn and other important agricultural commodities. In order to obtain nutrients from the host A. flavus produces a variety of extracellular hydrolytic enzymes. Int...

  14. Effects of Cymbopogon citratus L. essential oil on the growth, morphogenesis and aflatoxin production of Aspergillus flavus ML2-strain.

    PubMed

    Helal, G A; Sarhan, M M; Abu Shahla, A N K; Abou El-Khair, E K

    2007-02-01

    The mycelial growth of Aspergillus flavus Link was completely inhibited using 1.5 (microl/ml or 2.0 (microl/ml of Cymbopogon citratus essential oil applied by fumigation or contact method in Czapek's liquid medium, respectively. This oil was found also to be fungicidal at the same concentrations. The sublethal doses 1.0 and 1.5 (microl/ml inhibited about 65% of fungal growth after five days of incubation and delayed conidiation as compared with the control. Microscopic observations using Light Microscope (LM), Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) were carried out to determine the ultra structural modifications of A. flavus hyphae after treatment with C. citratus essential oil. The hyphal diameter decreased and hyphal wall appeared as precipitates and disappeared in some regions. This oil also caused plasma membrane disruption and mitochondrial structure disorganization. Moreover, Ca(+2), K(+) and Mg(+2) leakages increased from the fumigated mycelium and its total lipid content decreased, while the saturated and unsaturated fatty acids increased. One of the most important results obtained during this study was the ability of C. citratus essential oil at its sublethal dose to completely inhibit aflatoxin B(1) production from A. flavus. These findings increase the possibility of exploiting C. citratus essential oil as an effective inhibitor of biodegradation and storage contaminating fungi and also in fruit juice preservation. PMID:17304618

  15. Genomics of Aspergillus flavus mycotoxin production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aspergilli show immense ecological and metabolic diversity. To date, the sequences of fifteen different Aspergillus genomes have been determined providing scientists with an exciting resource to improve the understanding of Aspergillus molecular genomics. Aspergillus flavus, one of the most wide...

  16. Inhibition of growth of Aspergillus flavus and fungal alpha-amylases by a lectin-like protein from Lablab purpureus.

    PubMed

    Fakhoury, A M; Woloshuk, C P

    2001-08-01

    Aspergillus flavus is a fungal pathogen of maize causing an important ear rot disease when plants are exposed to drought and heat stress. Associated with the disease is the production of aflatoxins, which are a series of structurally related mycotoxins known to be carcinogenic. Previous research has suggested that the alpha-amylase of A. flavus promotes aflatoxin production in the endosperm of infected maize kernels. We report here the isolation and characterization of a 36-kDa alpha-amylase inhibitor from Lablab purpureus (AILP). AILP inhibited the alpha-amylases from several fungi but had little effect on those from animal and plant sources. The protein inhibited conidial germination and hyphal growth of A. flavus. The amino acid sequence indicated that AILP is similar to lectin members of a lectin-arcelin-alpha-amylase inhibitor family described in common bean and shown to be a component of plant resistance to insect pests. AILP also agglutinated papain-treated red blood cells from human and rabbit. These data indicate that AILP represents a novel variant in the lectin-arcelin-alpha-amylase inhibitor family of proteins having lectin-like and alpha-amylase inhibitory activity. PMID:11497467

  17. Recombination and cryptic heterokaryosis in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a pathogen of many agronomically important crops worldwide and can also cause human and animal diseases. A. flavus is the major producer of aflatoxins (AFs), which are carcinogenic secondary metabolites. In the United States, mycotoxins have been estimated to cause agricultur...

  18. Aspergillus flavus Genomics for Controlling Aflatoxin Contamination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The main objectives of the Aspergillus flavus genomics program are to identify genes and regulatory components involved in aflatoxin biosynthesis for solving aflatoxin contamination in agricultural crops. A. flavus Expressed Sequence Tags (EST), microarray and whole genome sequencing have been achi...

  19. Cinnamaldehyde inhibits fungal growth and aflatoxin B1 biosynthesis by modulating the oxidative stress response of Aspergillus flavus.

    PubMed

    Sun, Qi; Shang, Bo; Wang, Ling; Lu, Zhisong; Liu, Yang

    2016-02-01

    Cinnamaldehyde (CIN) is a promising natural preservative and generally recognized as safe for commodities as well as consumers. In this work, the antifungal effects of CIN on Aspergillus flavus were evaluated both in solid and in liquid culture conditions. Our results indicated that CIN effectively inhibited radial growth, spore production, mycelium formation, and aflatoxin B1 biosynthesis by A. flavus in a dose-dependent manner. At the concentration of 104 mg L(-1), CIN exposure was able to completely inhibit fungal growth as well as aflatoxin B1 production. Furthermore, the inhibitory activities of CIN were closely connected with the treatment period and the tested fungal species. Compared with the control strains, CIN dose dependently changed the morphology and ultrastructure of mycelium in different degree. Especially, the reduction of hydrogen peroxide was considered to follow the destruction of mitochondrial. Meanwhile, CIN significantly cut the levels of lipid peroxidation and reduced glutathione. The activity of total superoxide dismutase was significantly inhibited after CIN treatment at the end of incubation, whereas the activities of catalase and glutathione peroxidase were opposite. These results indicated that the inhibitory effect of CIN could attribute to oxidative stress alleviation possibly induced by modifications of cellular structure as well as redox status. PMID:26585445

  20. Mycotoxin production and predictive modelling kinetics on the growth of Aspergillus flavus and Aspergillus parasiticus isolates in whole black peppercorns (Piper nigrum L).

    PubMed

    Yogendrarajah, Pratheeba; Vermeulen, An; Jacxsens, Liesbeth; Mavromichali, Evangelia; De Saeger, Sarah; De Meulenaer, Bruno; Devlieghere, Frank

    2016-07-01

    The growth and mycotoxin production of three Aspergillus flavus isolates and an Aspergillus parasiticus isolate were studied in whole black peppercorns (Piper nigrum L.) using a full factorial design with seven water activity (aw) (0.826-0.984) levels and three temperatures (22, 30 and 37°C). Growth rates and lag phases were estimated using linear regression. Diverse secondary models were assessed for their ability to describe the radial growth rate as a function of individual and combined effect of aw and temperature. Optimum radial growth rate ranged from 0.75±0.04 to 2.65±0.02mm/day for A. flavus and 1.77±0.10 to 2.50±0.10mm/day for A. parasiticus based on the Rosso cardinal estimations. Despite the growth failure of some isolates at marginal conditions, all the studied models showed good performance to predict the growth rates. Validation of the models was performed on independently derived data. The bias factors (0.73-1.03), accuracy factors (0.97-1.36) and root mean square error (0.050-0.278) show that the examined models are conservative predictors of the colony growth rate of both fungal species in black peppers. The Rosso cardinal model can be recommended to describe the individual aw effect while the extended Gibson model was the best model for describing the combined effect of aw and temperature on the growth rate of both fungal species in peppercorns. Temperature optimum ranged from 30 to 33°C, while aw optimum was 0.87-0.92 as estimated by multi-factorial cardinal model for both species. The estimated minimum temperature and aw for A. flavus and A. parasiticus for growth were 11-16°C and 0.73-0.76, respectively, hence, achieving these conditions should be considered during storage to prevent the growth of these mycotoxigenic fungal species in black peppercorns. Following the growth study, production of mycotoxins (aflatoxins B1, B2, G1, G2, sterigmatocystin and O-methyl sterigmatocystin (OMST)) was quantified using LC-MS/MS. Very small

  1. Effect of Citrus reticulata and Cymbopogon citratus essential oils on Aspergillus flavus growth and aflatoxin production on Asparagus racemosus.

    PubMed

    Singh, Priyanka; Shukla, Ravindra; Kumar, Ashok; Prakash, Bhanu; Singh, Shubhra; Dubey, Nawal Kishore

    2010-09-01

    Essential oils extracted from Citrus reticulata and Cymbopogon citratus were tested in vitro against the toxigenic strain of Aspergillus flavus, isolated from the tuberous roots of Asparagus racemosus, used in preparation of herbal drugs. The essential oils completely inhibited the growth of A. flavus at 750 ppm and also exhibited a broad fungitoxic spectrum against nine additional fungi isolated from the roots. Citrus reticulata and Cymbopogon citratus essential oils completely inhibited aflatoxin B(1) production at 750 and 500 ppm, respectively. During in vivo investigation, the incidence of fungi and aflatoxin B(1) production decreased considerably in essential oil-treated root samples. The findings thus indicate possible exploitation of the essential oils as effective inhibitor of aflatoxin B(1) production and as post-harvest fungitoxicant of traditionally used plant origin for the control of storage fungi. These essential oils may be recommended as plant-based antifungals as well as aflatoxin B(1) suppressors in post-harvest processing of herbal samples. PMID:20401550

  2. Influence of propionic acid on growth and aflatoxin production by Aspergillus flavus in liquid submerged and solid substrate conditions.

    PubMed

    al-Hilli, A L; Smith, J E

    1992-01-01

    The present experiments demonstrate that sublethal concentrations of propionic acid stimulated aflatoxin production considerably in submerged shaken culture and solid substrate culture of Aspergillus flavus. In liquid conditions aflatoxin formation was significantly influenced by the time of addition of propionic acid. The spores initially swelled into large spherical cells, and the resultant hyphae developed into a swollen, stunted, and excessively branched mycelium. PMID:1573566

  3. Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1

    PubMed Central

    Farzaneh, Mohsen; Shi, Zhi-Qi; Ahmadzadeh, Masoud; Hu, Liang-Bin; Ghassempour, Alireza

    2016-01-01

    In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1), caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC) method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut. PMID:27298596

  4. Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1.

    PubMed

    Farzaneh, Mohsen; Shi, Zhi-Qi; Ahmadzadeh, Masoud; Hu, Liang-Bin; Ghassempour, Alireza

    2016-06-01

    In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1), caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC) method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut. PMID:27298596

  5. The effect of various antifungal agents on aflatoxin production and growth characteristics of Aspergillus parasiticus and Aspergillus flavus in liquid medium.

    PubMed

    Stewart, R G; Wyatt, R D; Ashmore, M D

    1977-09-01

    Various antifungal agents were added to a medium of 2% yeast extract-4% sucrose. Spores of a toxigenic strain of Aspergillus parasiticus were inoculated into the medium and incubated at 26 degrees C. for 10 days. Growth of the mold and aflatoxin formation were monitored every 48 hours during the experiment. Of the antifungal agents evaluated, propionic acid and crystal violet were the most effective compounds in retarding mold growth. Propionic acid was fungicidal at concentrations greater than 3.0 microgram/ml. whereas crystal violet exhibited a mold retarding activity at levels greater than 2,0 microgram/ml. Crystal violet retarded the growth rate of the mold during the initial stages of growth, however, this retardation was overcome after 10 days of incubation. Crystal violet also retarded aflatoxin production and sporulation of Aspergillus parasiticus; however, aflatoxin production was the most sensitive parameter. A survey involving 12 toxigenic isolates of A. parasiticus and A. flavus indicated that these species vary markedly in susceptibility to crystal violet. PMID:415299

  6. Regulation of Aspergillus flavus Aflatoxin Biosynthesis and Development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The filamentous fungus Aspergillus flavus produces a family of potent mutagenic and carcinogenic compounds collectively known as aflatoxins (AF). These secondary metabolites contaminate a number of oilseed crops during growth of the fungus and this can result in severe negative economic and health i...

  7. Evaluation of aflatoxin degradation by Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are highly toxic and hepatocarcinogenic compounds produced by Aspergillus flavus and A. parasiticus during infection of corn (maize), peanuts, cotton seed, and tree nuts (Figure 1). To minimize exposure to aflatoxins the U.S. Food and Drug Administration enforces a 20 ppb limit of aflatox...

  8. Genomic analysis of aspergillus flavus pathogenesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and Fusarium verticillioides colonize developing maize seeds and contaminate them with mycotoxins. Maize genotypes differ in resistance to these fungi, but incorporation of adequate resistance into desirable hybrids has been challenging.Little is known about pathogenesis of seeds...

  9. Interaction between maize seed and Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is an opportunistic fungal pathogen that colonizes maize seeds and contaminates them with aflatoxin. The fungus is localized in the endosperm and aleurone. To investigate the plant microbe interaction, we conducted histological and molecular studies to characterize the internal co...

  10. Cyclopiazonic acid biosynthesis by Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cyclopiazonic acid (CPA) is an indole-tetramic acid mycotoxin produced by some strains of Aspergillus flavus. Characterization of the CPA biosynthesis gene cluster confirmed that formation of CPA is via a three-enzyme pathway. This review examines the structure and organization of the CPA genes, elu...

  11. [Effect of alcoholic extracts of wild plants on the inhibition of growth of Aspergillus flavus, Aspergillus niger, Penicillium chrysogenum, Penicillium expansum, Fusarium moniliforme and Fusarium poae moulds].

    PubMed

    Tequida-Meneses, Martín; Cortez-Rocha, Mario; Rosas-Burgos, Ema Carina; López-Sandoval, Susana; Corrales-Maldonado, Consuelo

    2002-06-01

    Fungicidal activity of wild plants Larrea tridentata, Karwinskia humboldtiana, Ricinus communis, Eucalyptus globulus, Ambrosia ambrosioides, Nicotiana glauca, Ambrosia confertiflora, Datura discolor, Baccharis glutinosa, Proboscidea parviflora, Solanum rostratum, Jatropha cinerea, Salpianthus macrodonthus y Sarcostemma cynanchoides was evaluated against the moulds species Aspergillus flavus, Aspergillus niger, Penicillium chrysogenum, Penicillium expansum, Fusarium poae y Fusarium moniliforme moulds species. Alcoholic extracts 6% (w/v) were prepared using six grams of dried plant powders (leaves and stems) and alcohol (70% ethanol or 70% methanol). A spore suspension (1x10(6); ufc/ml) of each mould was prepared by adding saline solution (0.85%) and 0.1% tween 80. The extracts were mixed with Czapeck yeast agar (CYA) at 45-50 degrees C in 1:10 relation on Petri dishes. Triplicate Petri dishes of each treatment and for each mould were centrally inoculated and three Petri dishes were used without treatment as controls. The inoculated dishes and controls were incubated at 25 +/- 2 degrees C for eight days. The incubated dishes were examined each 48 h and after the colony diameter (radial growth) was measured. Two mould species were controlled by L. tridentata, B. glutinosa and P. parviflora. Extracts of L. tridentata in methanol or ethanol at 41.5-100% inhibited all six species of moulds. PMID:12828509

  12. Inhibition of aflatoxin metabolism and growth of Aspergillus flavus in liquid culture by a DNA methylation inhibitor.

    PubMed

    Yang, Kunlong; Zhuang, Zhenhong; Zhang, Feng; Song, Fengqin; Zhong, Hong; Ran, Fanlei; Yu, Song; Xu, Gaopo; Lan, Faxiu; Wang, Shihua

    2015-01-01

    Aflatoxins (AFs) are a group of highly oxygenated polyketidese-derived toxins mainly produced by Aspergillus flavus and A. parasiticus, whose biosynthesis mechanisms are extremely sophisticated. Methylation is known as the major form of epigenetic regulation, which is correlated with gene expression. As the DNA methylation inhibitor 5-azacytidine (5-AC) blocks AF production, we studied AFB1 metabolism and morphological changes of A. flavus by treatment with 5-AC in liquid culture. The results show that 5-AC caused a decrease in AF production and concurrent changes in morphology. In addition, we isolated a non-aflatoxigenic mutant of A. flavus, showing a significant reduction in pigment production, after 5-AC treatment. This mutant showed significant reduction in the expression of genes in the AF biosynthesis pathway, and conidia formation. Furthermore, as AF biosynthesis and oxidative stress are intimately related events, we assessed the viability of A. flavus to oxidative stress after treatment with 5-AC, which showed that the mutant was more sensitive to the strong oxidant hydrogen peroxide. We found that the non-aflatoxigenic mutant showed a decrease in reactive oxygen species (ROS) and metabolites indicative of oxidative stress, which may be caused by the disruption of the defence system against excessive ROS formation after 5-AC treatment. These data indicate that 5-AC, as an inactivator of DNA methyltransferase, plays a very important role in AFB1 metabolism and the development of A. flavus, which might provide an effective strategy to pre- or post-harvest control of AFs. PMID:25312249

  13. Aspergillus flavus SUMO Contributes to Fungal Virulence and Toxin Attributes.

    PubMed

    Nie, Xinyi; Yu, Song; Qiu, Mengguang; Wang, Xiuna; Wang, Yu; Bai, Youhuang; Zhang, Feng; Wang, Shihua

    2016-09-01

    Small ubiquitin-like modifiers (SUMOs) can be reversibly attached to target proteins in a process known as SUMOylation, and this process influences several important eukaryotic cell events. However, little is known regarding SUMO or SUMOylation in Aspergillus flavus. Here, we identified a novel member of the SUMO family in A. flavus, AfSumO, and validated the existence of SUMOylation in this pathogenic filamentous fungus. We investigated the roles of AfsumO in A. flavus by determining the effects of AfsumO mutations on the growth phenotype, stress response, conidia and sclerotia production, aflatoxin biosynthesis, and pathogenicity to seeds, and we found that SUMOylation plays a role in fungal virulence and toxin attributes. Taken together, these results not only reveal potential mechanisms of fungal virulence and toxin attributes in A. flavus but also provide a novel approach for promising new control strategies of this fungal pathogen. PMID:27532332

  14. Efficacy of aqueous garlic extract on growth, aflatoxin B1 production, and cyto-morphological aberrations of Aspergillus flavus, causing human ophthalmic infection: topical treatment of A. flavus keratitis.

    PubMed

    Ismaiel, Ahmed A; Rabie, Gamal H; Kenawey, Saied E M; Abd El-Aal, Marwa A

    2012-10-01

    By using agar well diffusion assay, antifungal activity of aqueous extract prepared from Egyptian garlic (Allium sativum L.) was evaluated in vitro against two strains of Aspergillus flavus (OC1 and OC10) causing human ocular infection. The recorded minimum inhibitory concentration (MIC) for growth inhibition of both strains was 3.60 mg/ml. Aqueous garlic extract (AGE) was used in successive in vivo tests as an attempt to cure rabbit's fungal keratitis caused by A. flavus OC1. Findings showed that diluted preparation of AGE was effective topical antifungal agent and succeeded to cure severe A. flavus keratitis in a time course less than 10 days without any observable side effects. Microscopic examination showed that AGE induced deleterious cyto-morphological aberrations in A. flavus target cells. AGE applied to Czapek's broth via contact method was more effective on growth, spores and aflatoxin B1 production than AGE applied to the same broth at the same concentration via fumigation method. PMID:24031964

  15. Cyclopiazonic Acid Biosynthesis of Aspergillus flavus and Aspergillus oryzae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cyclopiazonic acid (CPA) is an indole-tetramic acid neurotoxin produced by some of the same strains of A. flavus that produce aflatoxins and by some Aspergillus oryzae strains. Despite its discovery 40 years ago, few reviews of its toxicity and biosynthesis have been reported. This review examines w...

  16. Effects of temperature, water activity and incubation time on fungal growth and aflatoxin B1 production by toxinogenic Aspergillus flavus isolates on sorghum seeds.

    PubMed

    Lahouar, Amani; Marin, Sonia; Crespo-Sempere, Ana; Saïd, Salem; Sanchis, Vicente

    2016-01-01

    Sorghum, which is consumed in Tunisia as human food, suffers from severe colonization by several toxigenic fungi and contamination by mycotoxins. The Tunisian climate is characterized by high temperature and humidity that stimulates mold proliferation and mycotoxin accumulation in foodstuffs. This study investigated the effects of temperature (15, 25 and 37°C), water activity (aw, between 0.85 and 0.99) and incubation time (7, 14, 21 and 28 d) on fungal growth and aflatoxin B1 (AFB1) production by three Aspergillus flavus isolates (8, 10 and 14) inoculated on sorghum grains. The Baranyi model was applied to identify the limits of growth and mycotoxin production. Maximum diameter growth rates were observed at 0.99 a(w) at 37°C for two of the isolates. The minimum aw needed for mycelial growth was 0.91 at 25 and 37°C. At 15°C, only isolate 8 grew at 0.99 a(w). Aflatoxin B1 accumulation could be avoided by storing sorghum at low water activity levels (≤0.91 a(w)). Aflatoxin production was not observed at 15°C. This is the first work on the effects of water activity and temperature on A. flavus growth and AFB1 production by A. flavus isolates on sorghum grains. PMID:26920121

  17. Cyclopiazonic Acid Biosynthesis of Aspergillus flavus and Aspergillus oryzae

    PubMed Central

    Chang, Perng-Kuang; Ehrlich, Kenneth C.; Fujii, Isao

    2009-01-01

    Cyclopiazonic acid (CPA) is an indole-tetramic acid neurotoxin produced by some of the same strains of A. flavus that produce aflatoxins and by some Aspergillus oryzae strains. Despite its discovery 40 years ago, few reviews of its toxicity and biosynthesis have been reported. This review examines what is currently known about the toxicity of CPA to animals and humans, both by itself or in combination with other mycotoxins. The review also discusses CPA biosynthesis and the genetic diversity of CPA production in A. flavus/oryzae populations. PMID:22069533

  18. Nuclear heterogeneity in conidial populations of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a major producer of aflatoxin and an opportunistic pathogen for a wide range of hosts. Understanding genotypic and phenotypic variations within strains of A. flavus is important for controlling disease and reducing aflatoxin contamination. A. flavus is multinucleate and predomi...

  19. The potential role of oxidative stress in Aspergillus flavus survivability and aflatoxin biosynthesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination of food and feed occurs due to growth of Aspergillus flavus. This poses a serious health risk because of aflatoxin’s toxic and carcinogenic properties which negatively impact human and livestock health. Colonization and subsequent aflatoxin production by A. flavus is typicall...

  20. Identification of hydrolytic activities expressed by Aspergillus flavus grown on cotton carpel tissue

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus secreted at least two endoxylanase activities, two esterase activities and a pectolytic activity when grown on a medium containing cotton carpel tissue as a sole carbon source. A concentrated sample of A. flavus growth medium (6-day) was subjected to gel filtration chromatography...

  1. NON-TOXIGENIC ASPERGILLUS FLAVUS ISOLATES FOR REDUCING AFLATOXIN IN MISSISSIPPI DELTA CORN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The potential for two non-toxigenic isolates of Aspergillus flavus CT3 and K49 isolated from the Mississippi Delta to reduce aflatoxin contamination of corn was assessed in a field study. These two isolates exhibited comparable growth and aggressiveness as the toxigenic A. flavus isolate F3W4. The...

  2. A first glance into the genome sequence of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins, produced by Aspergillus flavus and A. parasiticus, are toxic and carcinogenic metabolites. They contaminate agricultural crops before harvest and post harvest grains during storage. In order to reduce and eliminate aflatoxin contamination of food and feed, Aspergillus flavus genomics p...

  3. Ecology, development and gene regulation in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is one of the most widely known species of Aspergillus. It was described as a species in 1809 and first reported as a plant pathogen in 1920. More recently, A. flavus has emerged as an important opportunistic pathogen and is now rec¬ognized as the second leading cause of aspergill...

  4. An attempt to model the probability of growth and aflatoxin B1 production of Aspergillus flavus under non-isothermal conditions in pistachio nuts.

    PubMed

    Aldars-García, Laila; Ramos, Antonio J; Sanchis, Vicente; Marín, Sonia

    2015-10-01

    Human exposure to aflatoxins in foods is of great concern. The aim of this work was to use predictive mycology as a strategy to mitigate the aflatoxin burden in pistachio nuts postharvest. The probability of growth and aflatoxin B1 (AFB1) production of aflatoxigenic Aspergillus flavus, isolated from pistachio nuts, under static and non-isothermal conditions was studied. Four theoretical temperature scenarios, including temperature levels observed in pistachio nuts during shipping and storage, were used. Two types of inoculum were included: a cocktail of 25 A. flavus isolates and a single isolate inoculum. Initial water activity was adjusted to 0.87. Logistic models, with temperature and time as explanatory variables, were fitted to the probability of growth and AFB1 production under a constant temperature. Subsequently, they were used to predict probabilities under non-isothermal scenarios, with levels of concordance from 90 to 100% in most of the cases. Furthermore, the presence of AFB1 in pistachio nuts could be correctly predicted in 70-81 % of the cases from a growth model developed in pistachio nuts, and in 67-81% of the cases from an AFB1 model developed in pistachio agar. The information obtained in the present work could be used by producers and processors to predict the time for AFB1 production by A. flavus on pistachio nuts during transport and storage. PMID:26187836

  5. Suppression of Aflatoxin Biosynthesis in Aspergillus flavus by 2-Phenylethanol Is Associated with Stimulated Growth and Decreased Degradation of Branched-Chain Amino Acids.

    PubMed

    Chang, Perng-Kuang; Hua, Sui Sheng T; Sarreal, Siov Bouy L; Li, Robert W

    2015-10-01

    The saprophytic soil fungus Aspergillus flavus infects crops and produces aflatoxin. Pichia anomala, which is a biocontrol yeast and produces the major volatile 2-phenylethanol (2-PE), is able to reduce growth of A. flavus and aflatoxin production when applied onto pistachio trees. High levels of 2-PE are lethal to A. flavus and other fungi. However, at low levels, the underlying mechanism of 2-PE to inhibit aflatoxin production remains unclear. In this study, we characterized the temporal transcriptome response of A. flavus to 2-PE at a subinhibitory level (1 μL/mL) using RNA-Seq technology and bioinformatics tools. The treatment during the entire 72 h experimental period resulted in 131 of the total A. flavus 13,485 genes to be significantly impacted, of which 82 genes exhibited decreased expression. They included those encoding conidiation proteins and involved in cyclopiazonic acid biosynthesis. All genes in the aflatoxin gene cluster were also significantly decreased during the first 48 h treatment. Gene Ontology (GO) analyses showed that biological processes with GO terms related to catabolism of propionate and branched-chain amino acids (valine, leucine and isoleucine) were significantly enriched in the down-regulated gene group, while those associated with ribosome biogenesis, translation, and biosynthesis of α-amino acids OPEN ACCESS Toxins 2015, 7 3888 were over-represented among the up-regulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that metabolic pathways negatively impacted among the down-regulated genes parallel to those active at 30 °C, a condition conducive to aflatoxin biosynthesis. In contrast, metabolic pathways positively related to the up-regulated gene group resembled those at 37 °C, which favors rapid fungal growth and is inhibitory to aflatoxin biosynthesis. The results showed that 2-PE at a low level stimulated active growth of A. flavus but concomitantly rendered decreased activities in

  6. Suppression of Aflatoxin Biosynthesis in Aspergillus flavus by 2-Phenylethanol Is Associated with Stimulated Growth and Decreased Degradation of Branched-Chain Amino Acids

    PubMed Central

    Chang, Perng-Kuang; Hua, Sui Sheng T.; Sarreal, Siov Bouy L.; Li, Robert W.

    2015-01-01

    The saprophytic soil fungus Aspergillus flavus infects crops and produces aflatoxin. Pichia anomala, which is a biocontrol yeast and produces the major volatile 2-phenylethanol (2-PE), is able to reduce growth of A. flavus and aflatoxin production when applied onto pistachio trees. High levels of 2-PE are lethal to A. flavus and other fungi. However, at low levels, the underlying mechanism of 2-PE to inhibit aflatoxin production remains unclear. In this study, we characterized the temporal transcriptome response of A. flavus to 2-PE at a subinhibitory level (1 µL/mL) using RNA-Seq technology and bioinformatics tools. The treatment during the entire 72 h experimental period resulted in 131 of the total A. flavus 13,485 genes to be significantly impacted, of which 82 genes exhibited decreased expression. They included those encoding conidiation proteins and involved in cyclopiazonic acid biosynthesis. All genes in the aflatoxin gene cluster were also significantly decreased during the first 48 h treatment. Gene Ontology (GO) analyses showed that biological processes with GO terms related to catabolism of propionate and branched-chain amino acids (valine, leucine and isoleucine) were significantly enriched in the down-regulated gene group, while those associated with ribosome biogenesis, translation, and biosynthesis of α-amino acids were over-represented among the up-regulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that metabolic pathways negatively impacted among the down-regulated genes parallel to those active at 30 °C, a condition conducive to aflatoxin biosynthesis. In contrast, metabolic pathways positively related to the up-regulated gene group resembled those at 37 °C, which favors rapid fungal growth and is inhibitory to aflatoxin biosynthesis. The results showed that 2-PE at a low level stimulated active growth of A. flavus but concomitantly rendered decreased activities in branched-chain amino acid degradation

  7. Removal of aflatoxin B1 and inhibition of Aspergillus flavus growth by the use of Lactobacillus plantarum on olives.

    PubMed

    Kachouri, Faten; Ksontini, Hamida; Hamdi, Moktar

    2014-10-01

    Olives can be contaminated with a wide variety of molds (Aspergillus and/or Penicillium) that can be occurring naturally on fresh and processed olives and could support mycotoxin production. The aim of this work was to investigate aflatoxin B1 (AFB1) production by fungi and its bioaccumulation in olives during storage and to study the impact of the application of Lactobacillus plantarum on the inhibition of mold development and production of AFB1. Two different treatments were applied: (i) olives with natural microflora and (ii) olives inoculated with Aspergillus flavus after elimination of natural microflora. AFB1 has been extracted from olives and quantitated by high-performance liquid chromatography using a fluorescence detector. Results showed the absence of this metabolite in the olives for the season 2008 to 2009. In 2009 to 2010, AFB1 was detected at the level of 11 μg/kg. The application of L. plantarum during the storage of olives favors the reduction of the level of AFB1 to 5.9 μg/kg correlated with a decrease in the amount of molds (86.3%). The images obtained by environmental scanning electron microscopy showed that L. plantarum was able to adhere to the olive surface and probably produce a biofilm that inhibits the multiplication of yeast and fungi by oxygen competition. Results showed an increase of antioxidant activity and amount of total phenolic compounds of olives, respectively, by 24 and 8.6%. In many olives contaminated with A. flavus, AFB1 was present at an initial level of 5.15 μg/kg and increased to 6.55 μg/kg after 8 days of storage. The biological detoxification of AFB1 in olives by L. plantarum is confirmed by the reduction of the level of AFB1 to 2.12 μg/kg on day 0 and its absence after 4 days of storage. PMID:25285494

  8. The major volatile compound 2-phenylethanol from the biocontrol yeast, Pichia anomala, inhibits growth and expression of aflatoxin biosynthetic genes of Aspergillus flavus.

    PubMed

    Hua, Sui Sheng T; Beck, John J; Sarreal, Siov Bouy L; Gee, Wai

    2014-05-01

    Aspergillus flavus is a ubiquitous saprophyte that is able to produce the most potent natural carcinogenic compound known as aflatoxin B1 (AFB1). This toxin frequently contaminates crops including corn, cotton, peanuts, and tree nuts causing substantial economic loss worldwide. Consequently, more than 100 countries have strict regulations limiting AFB1 in foodstuffs and feedstuffs. Plants and microbes are able to produce volatile compounds that act as a defense mechanism against other organisms. Pichia anomala strain WRL-076 is a biocontrol yeast currently being tested to reduce AF contamination of tree nuts in California. We used the SPME-GC/MS analysis and identified the major volatile compound produced by this strain to be 2-phenylethanol (2-PE). It inhibited spore germination and AF production of A. flavus. Inhibition of AF formation by 2-PE was correlated with significant down regulation of clustering AF biosynthesis genes as evidenced by several to greater than 10,000-fold decrease in gene expression. In a time-course analysis we found that 2-PE also altered the expression patterns of chromatin modifying genes, MYST1, MYST2, MYST3, gcn5, hdaA and rpdA. The biocontrol capacity of P. anomala can be attributed to the production of 2-PE, which affects spore germination, growth, toxin production, and gene expression in A. flavus. PMID:24504634

  9. Scleral Buckle Infection with Aspergillus Flavus

    PubMed Central

    Bouhaimed, Manal; Al-Dhibi, Hassan; Al-Assiri, Abdullah

    2008-01-01

    Purpose: To present a case of scleral buckle infection with Aspergillus flavus in a tertiary eye center in Saudi Arabia. Methods: A retrospective case report of a 28-year-old Saudi male who presented with a six-month history of conjunctival injection and discharge from the left eye which had undergone uncomplicated conventional retinal detachment surgery, at the King Khaled Eye Specialist Hospital in Riyadh, Saudi Arabia, in the form of cryopexy, subretinal fluid drainage and scleral buckle (grooved segmental sponge and circumferential band with sleeve) for a macula on retinal detachment four years earlier. A diagnosis of infected extruded scleral buckle was made and the buckle was removed. Results: The infected scleral buckle was removed under local anesthesia with administration of sub-conjunctival irrigation of 50 mg solution of Vancomycin, and sub-conjunctival injection of 25mg of Vancomycin. Post operative microbiological studies revealed infection with silver staining of moderate Aspergillus flavus hyphae. Visual acuity of the left eye improved from 20/200 before surgery to 20/60 in the two years follow-up visit. Conclusion: This case report indicates the importance of considering infection with multiple organisms – including fungal ones – in cases of scleral buckle infections in our population. PMID:20379425

  10. Effect of Equisetum arvense and Stevia rebaudiana extracts on growth and mycotoxin production by Aspergillus flavus and Fusarium verticillioides in maize seeds as affected by water activity.

    PubMed

    Garcia, Daiana; Ramos, Antonio J; Sanchis, Vicente; Marín, Sonia

    2012-02-01

    Cereals are a very important part of the human and animal diets. However, agricultural products can be contaminated by moulds and their mycotoxins. Plant extracts, particularly those of Equisetum arvense and Stevia rebaudiana have been reported previously to contain antioxidant compounds which may have antifungal properties. In this study, E. arvense and S. rebaudiana extracts were tested for their control of mycotoxigenic fungi in maize. The extracts were tested separately and as a mixture for their effect on growth of Aspergillus flavus and Fusarium verticillioides. Extracts were added to unsterilised inoculated maize at different water activity (a(w)) levels (0.85-0.95). Moulds were inoculated and incubated for 30 days. Results confirmed that the extract of E. arvense and a mixture 1:1 of Equisetum-Stevia may be effective for the inhibition of both growth of A. flavus and aflatoxin production at high water activity levels (pre-harvest conditions). In general, growth of the F. verticillioides was reduced by the use of plant extracts, especially at 0.95 a(w). However, fumonisin presence was not significantly affected. E. arvense and S. rebaudiana extracts could be developed as an alternative treatment to control aflatoxigenic mycobiota in moist maize. PMID:22104120

  11. Occurrence of aflatoxins in mahua (Madhuca indica Gmel.) seeds: synergistic effect of plant extracts on inhibition of Aspergillus flavus growth and aflatoxin production.

    PubMed

    Sidhu, O P; Chandra, Harish; Behl, H M

    2009-04-01

    Occurrence of aflatoxin in Madhuca indica Gmel. seeds was determined by competitive ELISA. Eighty percent of mahua seed samples were found to be contaminated with aflatoxin. Total aflatoxin content ranged from 115.35 to 400.54ppb whereas the concentration of AFB(1) was in the range of 86.43 to 382.45ppb. Mahua oil was extracted by cold press expeller and analysed for contamination of aflatoxin in both the oil and cake samples. Total aflatoxin and aflatoxin B(1) were 220.66 and 201.57ppb in oil as compared to that in cake samples where it was 87.55 and 74.35ppb, respectively. Various individual and combined plant extracts were evaluated for their efficacy against growth of Aspergillus flavus and aflatoxin production in vitro. Combination of botanicals were found to be more effective in controlling fungal growth and aflatoxin production than individual extracts. Results of the present study suggests that synergistic effect of plant extracts can be used for control of fungal growth and aflatoxin production. These natural plant products may successfully replace synthetic chemicals and provide an alternative method to protect mahua as well as other agricultural commodities of nutritional significance from toxigenic fungi such as A. flavus and aflatoxin production. PMID:19167450

  12. Nutrient environment influences competition among Aspergillus flavus genotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Structures of Aspergillus flavus populations, shaped by intraspecific competition, influence the incidences and severities of crop aflatoxin contamination. Competition for nutrients may be one factor modulating intraspecific interactions, but influences of specific types and concentrations of nutrie...

  13. Population shifts and mating-type heterokaryosis in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a fungal pathogen of many agronomically important crops worldwide. We sampled A. flavus strains from a cornfield in Rocky Mount, NC. This field was planted in 2010 and plots were inoculated at tasselling with either AF36 or NRRL 21882 (=Afla-Guard) biocontrol strains, both of...

  14. Mating-type heterokaryosis and population shifts in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a fungal pathogen of many agronomically important crops worldwide. We sampled A. flavus strains from a cornfield in Rocky Mount, NC. This field was planted in 2010 and plots were inoculated at tasselling with either AF36 or NRRL 21882 (=Afla-Guard) biocontrol strains, both of...

  15. The maize rachis affects Aspergillus flavus movement during ear development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus expressing green fluorescent protein (GFP) was used to follow infection in ears of maize hybrids resistant and susceptible to the fungus. Developing ears were needle-inoculated with GFP-transformed A. flavus 20 days after silk emergence, and GFP fluorescence in the pith was evalu...

  16. Sexual reproduction in Aspergillus flavus sclerotia naturally produced in corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is the major producer of carcinogenic aflatoxins worldwide in crops. Populations of A. flavus are characterized by high genetic variation and the source of this variation is likely sexual reproduction. The fungus is heterothallic and laboratory crosses produce ascospore-bearing ...

  17. Population structure of Aspergillus flavus before and after biocontrol treatment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a fungal pathogen of many important crops worldwide. We sampled A. flavus strains from a cornfield in Rocky Mount, North Carolina, over a period of two years. Plots were inoculated at tasselling with either AF36 or NRRL 21882 (=Afla-Guard) biocontrol strains, both of which are ...

  18. Population dynamics of Aspergillus flavus following biocontrol treatment of corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a fungal pathogen of many agronomically important crops worldwide. We sampled A. flavus strains from a cornfield in Rocky Mount, North Carolina, over a period of two years. The field was planted in 2010 and plots were inoculated at tasselling with either AF36 or NRRL 21882 (=Af...

  19. Potential of Aspergillus flavus Genomics for Applications in Biotechnology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a common saprophyte and opportunistic pathogen that survives in the natural environment by extracting nutrition from plant debris, insect carcasses and a variety of other carbon sources. A. flavus produces numerous secondary metabolites and hydrolytic enzymes. The primary obj...

  20. Population ecology of Aspergillus flavus associated with Mississippi Delta Soils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Understanding the source of Aspergillus flavus is required to effectively manage within-field aflatoxin contamination of maize (Zea mays L.). Studies assessed the density of A. flavus propagules and other soil microflora (Fusarium spp., total fungi) associated with Mississippi Delta soils, and corr...

  1. The major volatile compound 2-phenylethanol from the biocontrol yeast Pichia anomala inhibits growth and expression of aflatoxin biosynthetic genes of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a globally distributed fungus and an important food contaminant because it produces the most potent natural carcinogenic compound known as aflatoxin (AF) B1. The major volatile from a yeast strain, Pichia anomala WRL-076 was identified by SPEM-GC/MS analysis to be 2-phenylethan...

  2. What can Aspergillus flavus genome offer for mycotoxin research?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genomic study of filamentous fungi has made significant advances in recent years, and the genomes of several species in the genus Aspergillus have been sequenced, including Aspergillus flavus. This ubiquitous mold is present as a saprobe in a wide range of agricultural and natural habits, and c...

  3. WHOLE GENOME COMPARISON OF ASPERGILLUS FLAVUS AND A. ORYZAE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a plant and animal pathogen that also produces the potent carcinogen aflatoxin. Aspergillus oryzae is a closely related species that has been used for centuries in the food fermentation industry and is generally regarded as safe (GRAS). Whole genome sequences for these two fu...

  4. Nitrification of Aspartate by Aspergillus flavus

    PubMed Central

    Hatcher, H. J.; Schmidt, E. L.

    1971-01-01

    Heterotrophic conversion of l-aspartic acid to nitrification products by Aspergillus flavus was studied in a replacement incubation system. Numerous amino acids supported nitrification; aspartate and glutamate were about equivalent as the best sources of nitrate. Addition of sodium bicarbonate to the incubation system substantially enhanced nitrate formation for all nitrifiable amino acids except aspartic acid, but the basis for the bicarbonate effect is obscure. The yield of nitrate from l-aspartate was not approached by forms of aspartic acid resulting from substitution on the beta carbon, the amino nitrogen, or the gamma carboxyl group or by aspartate presented as the d-configuration. There was no relationship between nitrate formation and the occurrence of such possible intermediates as nitrite, bound hydroxylamine, ammonia, aspergillic acid, and beta-nitropropionic acid. Uniformly labeled 14C-l-aspartate that was nitrified in replacement incubation led to no accumulation of label in possible nitrification products in the culture filtrate. Label was found in components of the mycelium after acid hydrolysis, with heaviest accumulation in what appeared to be glucosamine and an unidentified compound, possibly acetylglucosamine. Detectable label was redistributed into serine, glycine, and threonine. Images PMID:5549699

  5. Enhanced diversity and aflatoxigenicity in interspecific hybrids of Aspergillus flavus and Aspergillus parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and A. parasiticus are two of the most important aflatoxin-producing species that contaminate agricultural commodities worldwide. Both species are heterothallic and undergo sexual reproduction in laboratory crosses. Here, we examine the possibility of interspecific matings betwe...

  6. Functional Genomic Analysis of Aspergillus flavus Interacting with Resistant and Susceptible Peanut

    PubMed Central

    Wang, Houmiao; Lei, Yong; Yan, Liying; Wan, Liyun; Ren, Xiaoping; Chen, Silong; Dai, Xiaofeng; Guo, Wei; Jiang, Huifang; Liao, Boshou

    2016-01-01

    In the Aspergillus flavus (A. flavus)–peanut pathosystem, development and metabolism of the fungus directly influence aflatoxin contamination. To comprehensively understand the molecular mechanism of A. flavus interaction with peanut, RNA-seq was used for global transcriptome profiling of A. flavus during interaction with resistant and susceptible peanut genotypes. In total, 67.46 Gb of high-quality bases were generated for A. flavus-resistant (af_R) and -susceptible peanut (af_S) at one (T1), three (T2) and seven (T3) days post-inoculation. The uniquely mapped reads to A. flavus reference genome in the libraries of af_R and af_S at T2 and T3 were subjected to further analysis, with more than 72% of all obtained genes expressed in the eight libraries. Comparison of expression levels both af_R vs. af_S and T2 vs. T3 uncovered 1926 differentially expressed genes (DEGs). DEGs associated with mycelial growth, conidial development and aflatoxin biosynthesis were up-regulated in af_S compared with af_R, implying that A. flavus mycelia more easily penetrate and produce much more aflatoxin in susceptible than in resistant peanut. Our results serve as a foundation for understanding the molecular mechanisms of aflatoxin production differences between A. flavus-R and -S peanut, and offer new clues to manage aflatoxin contamination in crops. PMID:26891328

  7. Population ecology of Aspergillus flavus associated with Mississippi Delta soils.

    PubMed

    Zablotowicz, R M; Abbas, H K; Locke, M A

    2007-10-01

    Understanding the source of Aspergillus flavus is required to manage aflatoxin contamination of maize (Zea mays L.). Studies assessed A. flavus propagules, Fusarium spp., and total fungi associated with Mississippi Delta soils. Soils from 12 and 15 sites were collected in 2000 and 2001, respectively. The propagule density of A. flavus ranged from log(10) 2.0 to 4.3 colony-forming units (cfu) g(-1) soil, while total fusaria ranged from log(10) 3.0 to 5.4 cfu g(-1) soil. The highest populations of A. flavus were associated with soils containing higher organic matter, especially in sites under a no-tillage management. The frequency of aflatoxin production in isolates ranged from 13 to 81% depending on soil. In 2001, there was a highly significant correlation between A. flavus and the history of maize cultivation. Soil fertility factors such as organic matter content, nitrate and extractable phosphorus correlated with the density of Aspergillus, Fusarium spp., and total fungi. The relationship between soil parameters and Aspergillus populations may be useful in predicting the contribution of soil microflora to aflatoxin contamination. PMID:17886181

  8. Nonaflatoxigenic Aspergillus flavus TX9-8 Competitively Prevents Aflatoxin Production by A. flavus Isolates of Large and Small Sclerotial Morphotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Toxigenic Aspergillus flavus is the main etiological agent for aflatoxin contamination of crops. Using nonaflatoxigenic A. flavus isolates to competitively exclude toxigenic A. flavus isolates in agricultural fields has become an adopted approach to reduce aflatoxin contamination. We determined th...

  9. Mating-type heterokaryosis in Aspergillus flavus in North Carolina

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a well-known pathogen of many important agricultural commodities and is a major producer of aflatoxins (AFs), which are carcinogenic polyketides that pose a serious health risk to humans and animals. Recently, heterokaryosis and the presence of cryptic alleles were shown to ex...

  10. Genetic Response to Seed Colonizatin by Aspergillus flavus in Peanut

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous studies to evaluate peanut genotypes for in vitro resistance to seed colonization by Aspergillus flavus have not resulted in the development of cultivars with resistance to aflatoxin contamination in the field. New breeding lines showing pre-harvest field resistance to aflatoxin contaminat...

  11. Evidence of aneuploidy modulating aflatoxigenicity in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a well-known pathogen of many important agricultural commodities and is a major producer of aflatoxins, which are carcinogenic polyketides that pose a serious health risk to humans and animals. Aflatoxin contamination in peanut exports worldwide accounts for as much as $450 mi...

  12. Sexual reproduction in Aspergillus flavus and A. parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and A. parasiticus are potent producers of carcinogenic and hepatotoxic aflatoxins, polyketide-derived secondary metabolites that contaminate a wide variety of agricultural crops. Strains with opposite mating-type genes MAT1-1 and MAT1-2 within each species were crossed in an att...

  13. Cryptic Sexuality in Aspergillus parasiticus and A. flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascomycetous fungi of the genus Aspergillus comprise a wide variety of species of biotechnological importance (e.g. A. sojae, A. oryzae, A. niger) as well as pathogens and toxin producers (e.g. A. flavus, A. parasiticus, A. fumigatus, A. nidulans). With the exception of A. nidulans, which is a homot...

  14. Characterization of toxigenic and atoxigenic Aspergillus flavus isolates from pistachio

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thirty eight Aspergillus flavus isolates collected from a pistachio orchard in California were analyzed for production of aflatoxin (AF), cyclopiazonic acid (CPA), vegetative compatibility groups (VCGs) and mating types. All toxigenic isolates produced both AFB1 and CPA. Twenty-one percent of the i...

  15. METHODS TO SAMPLE AIR BORNE PROPAGULES OF ASPERGILLUS FLAVUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several techniques (cyclone samplers, filter samplers and rotorods) were evaluated for detection of airborne Aspergillus flavus Link propagules in a cultivated region of southwest Arizona. Cyclone samplers operated continuously for 168 h (7 d) collected a dry sample that was ideal for quantificatio...

  16. Recombination and cryptic heterokaryosis in experimental populations of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus infects both plants and animals, and is of toxicological importance due to its production of aflatoxins (AFs) and other mycotoxins. Mycotoxins can cause agricultural losses totaling upwards of $1.4 billion annually. Recent efforts to reduce AF concentrations have focused on the us...

  17. The effect of 2-phenylethanol treatment on Aspergillus flavus transcriptome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pichia anomalais, which produces the antimicrobial volatile 2-phenylethanol (2-PE), is effective in reducing A. flavus growth and aflatoxin production. We treated A. flavus NRRL3357 with 2-PE and analyzed changes in the transcriptomic profiles at different stages of fungal growth. RNA-Seq reads from...

  18. Molecular characterization of atoxigenic Aspergillus flavus isolates collected in China.

    PubMed

    Wei, Dandan; Zhou, Lu; Selvaraj, Jonathan Nimal; Zhang, Chushu; Xing, Fuguo; Zhao, Yueju; Wang, Yan; Liu, Yang

    2014-07-01

    Aspergillus flavus strains were isolated from peanut fields of Liaoning, Shandong, Hubei and Guangdong Provinces in China, and identified through phenotypic and molecular approaches. Of the 323 A. flavus strains isolated, 76 strains did not produce aflatoxins detectable by UPLC. The incidence of atoxigenic A. flavus strains decreased with increase in temperature and increased with increase in latitude in different geographical locations. Amplification of all the aflatoxin genes in the aflatoxin gene cluster in the atoxigenic isolates showed that there were 25 deletion patterns (A-Y), with 22 deletion patterns identified for the first time. Most of the atoxigenic A. flavus isolates with gene deletions (97%) had deletions in at least one of the four genes (aflT, nor-1, aflR, and hypB), indicating that these four genes could be targeted for rapid identification of atoxigenic strains. The atoxigenic isolates with gene deletions, especially the isolates with large deletions, are potential candidates for aflatoxin control. PMID:24879349

  19. Environmental and Developmental Factors Influencing Aflatoxin Production by Aspergillus flavus and Aspergillus parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are carcinogenic mycotoxins formed by a number of fungi in the genus Aspergillus. The major fungi responsible for aflatoxin formation in crop seeds in the field and in storage are Aspergillus flavus and A. parasiticus. This review emphasizes developmental, environmental, biological and ...

  20. Hyperspectral imagery for observing spectral signature change in Aspergillus flavus

    NASA Astrophysics Data System (ADS)

    DiCrispino, Kevin; Yao, Haibo; Hruska, Zuzana; Brabham, Kori; Lewis, David; Beach, Jim; Brown, Robert L.; Cleveland, Thomas E.

    2005-11-01

    Aflatoxin contaminated corn is dangerous for domestic animals when used as feed and cause liver cancer when consumed by human beings. Therefore, the ability to detect A. flavus and its toxic metabolite, aflatoxin, is important. The objective of this study is to measure A. flavus growth using hyperspectral technology and develop spectral signatures for A. flavus. Based on the research group's previous experiments using hyperspectral imaging techniques, it has been confirmed that the spectral signature of A. flavus is unique and readily identifiable against any background or surrounding surface and among other fungal strains. This study focused on observing changes in the A. flavus spectral signature over an eight-day growth period. The study used a visible-near-infrared hyperspectral image system for data acquisition. This image system uses focal plane pushbroom scanning for high spatial and high spectral resolution imaging. Procedures previously developed by the research group were used for image calibration and image processing. The results showed that while A. flavus gradually progressed along the experiment timeline, the day-to-day surface reflectance of A. flavus displayed significant difference in discreet regions of the wavelength spectrum. External disturbance due to environmental changes also altered the growth and subsequently changed the reflectance patterns of A. flavus.

  1. Potential involvement of Aspergillus flavus laccases in peanut invasion at low water potential

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus (Link) accumulates aflatoxins in peanuts, mainly affecting immature kernels during drought. Peanut invasion by A. flavus induces synthesis of phytoalexins, mostly stilbenoids, as a plant defense mechanism. Fungal laccases are often related to pathogenicity, and among other subst...

  2. Genes Differentially Expressed by Aspergillus flavus Strains After Loss of Aflatoxin Production by Serial Transfers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are carcinogenic fungal secondary metabolites produced by Aspergillus flavus and other closely related species. To better understand the molecular events that are associated with aflatoxin production, three separate nonaflatoxigenic A. flavus strains were produced through serial transfer...

  3. Comparative Genomics of Aspergillus flavus and A. oryzae: An Early View

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus produces aflatoxins and is the second leading cause of aspergillosis in immunocompromised individuals. Aspergillus oryzae, on the other hand, has been used for centuries in Japan for the fermentation of food. The recently available whole genome sequences of Aspergillus flavus an...

  4. Lipids in Aspergillus flavus-maize interaction

    PubMed Central

    Scarpari, Marzia; Punelli, Marta; Scala, Valeria; Zaccaria, Marco; Nobili, Chiara; Ludovici, Matteo; Camera, Emanuela; Fabbri, Anna A.; Reverberi, Massimo; Fanelli, Corrado

    2014-01-01

    In some filamentous fungi, the pathways related to the oxidative stress and oxylipins production are involved both in the process of host-recognition and in the pathogenic phase. In fact, recent studies have shown that the production of oxylipins in filamentous fungi, yeasts and chromists is also related to the development of the organism itself and to mechanisms of communication with the host at the cellular level. The oxylipins, also produced by the host during defense reactions, are able to induce sporulation and to regulate the biosynthesis of mycotoxins in several pathogenic fungi. In A. flavus, the oxylipins play a crucial role as signals for regulating the biosynthesis of aflatoxins, the conidiogenesis and the formation of sclerotia. To investigate the involvement of an oxylipins based cross-talk into Z. mays and A. flavus interaction, we analyzed the oxylipins profile of the wild type strain and of three mutants of A. flavus that are deleted at the Aflox1 gene level also during maize kernel invasion. A lipidomic approach has been addressed through the use of LC-ToF-MS, followed by a statistical analysis of the principal components (PCA). The results showed the existence of a difference between the oxylipins profile generated by the WT and the mutants onto challenged maize. In relation to this, aflatoxin synthesis which is largely hampered in vitro, is intriguingly restored. These results highlight the important role of maize oxylipin in driving secondary metabolism in A. flavus. PMID:24578700

  5. Inhibition of Aflatoxin Production of Aspergillus flavus by Lactobacillus casei

    PubMed Central

    Chang, Injeong

    2007-01-01

    Lactobacillus casei KC-324 was tested for its ability to inhibit aflatoxin production and mycelial growth of Aspergillus flavus ATCC 15517 in liquid culture. Aflatoxin B1 biosynthesis and mycelial growth were inhibited in both simultaneous culture and individual antagonism assays,suggesting that the inhibitory activity was due to extracellular metabolites produced in cell-free supernatant fluids of the cultured broth of L. casei KC-324. In cell-free supernatant fluids of all media tested,deMan,Rogosa and Sharpe broth,potato dextrose broth,and Czapek-Dox broth + 1% yeast extract showed higher antiaflatoxigenic activity. In these case, fungal growths, however, was not affected as measured by mycelial dry weight. The antiaflatoxigenic metabolites from L. casei KC-324 were produced over wide range of temperatures between 25℃ and 37℃. However, these metabolites were not thermostable since the inhibitory activity of the supernatant was inactivated within 30 minutes at 100℃ and 121℃. The inhibitory activity was not influenced by changing pH of supernatant between 4 and 10. However,the antiaflatoxigenic activity was slightly reduced at pH 10. PMID:24015075

  6. The complete mitochondrial genome sequence of Aspergillus flavus.

    PubMed

    Yan, Zhengsong; Chen, Dan; Shen, Yiping; Ye, Baodong

    2016-07-01

    Aspergillus flavus is a haploid filamentous fungus that is common in the environment and has been implicated in human infections. The complete mitochondrial genome of A. flavus has been determined by high-throughput sequencing technology in this work. Our study revealed that the mitochondrial genome of A. flavus is 31,602 bp long, with an A + T content of 74.83%, which consists of a usual set of mitochondrial proteins and RNA genes, including large and small ribosomal RNAs, 15 proteins, and 20 tRNA genes and contains two introns. Notably, it also contains two hypothetical proteins without obvious homology to any known proteins. All structural genes are located on one strand and are apparently transcribed in one direction. Codon usage analysis indicated that all protein coding genes employ the standard fungal mitochondrial start and stop codons; and the nucleotide bias toward AT was also reflected in the codon usage. The complete mitochondrial genomes of A. flavus would be useful for future investigation of the genetic, evolution, and clinical identification of Aspergillus species. PMID:25922962

  7. A systems approach to model the relationship between aflatoxin gene cluster expression, environmental factors, growth and toxin production by Aspergillus flavus

    PubMed Central

    Abdel-Hadi, Ahmed; Schmidt-Heydt, Markus; Parra, Roberto; Geisen, Rolf; Magan, Naresh

    2012-01-01

    A microarray analysis was used to examine the effect of combinations of water activity (aw, 0.995–0.90) and temperature (20–42°C) on the activation of aflatoxin biosynthetic genes (30 genes) in Aspergillus flavus grown on a conducive YES (20 g yeast extract, 150 g sucrose, 1 g MgSO4·7H2O) medium. The relative expression of 10 key genes (aflF, aflD, aflE, aflM, aflO, aflP, aflQ, aflX, aflR and aflS) in the biosynthetic pathway was examined in relation to different environmental factors and phenotypic aflatoxin B1 (AFB1) production. These data, plus data on relative growth rates and AFB1 production under different aw × temperature conditions were used to develop a mixed-growth-associated product formation model. The gene expression data were normalized and then used as a linear combination of the data for all 10 genes and combined with the physical model. This was used to relate gene expression to aw and temperature conditions to predict AFB1 production. The relationship between the observed AFB1 production provided a good linear regression fit to the predicted production based in the model. The model was then validated by examining datasets outside the model fitting conditions used (37°C, 40°C and different aw levels). The relationship between structural genes (aflD, aflM) in the biosynthetic pathway and the regulatory genes (aflS, aflJ) was examined in relation to aw and temperature by developing ternary diagrams of relative expression. These findings are important in developing a more integrated systems approach by combining gene expression, ecophysiological influences and growth data to predict mycotoxin production. This could help in developing a more targeted approach to develop prevention strategies to control such carcinogenic natural metabolites that are prevalent in many staple food products. The model could also be used to predict the impact of climate change on toxin production. PMID:21880616

  8. Aspergillus flavus-Induced Brain Abscess in an Immunocompetent Child

    PubMed Central

    Al-Maskari, Nawal; Hussain, Ibrahim; Jumaa, Suleiman; Al-Shail, Essam A.

    2016-01-01

    Intracranial aspergillosis is an extremely rare manifestation of invasive aspergillosis in immunocompetent children and is associated with high morbidity and mortality. We report a 12-year-old immunocompetent male child who was referred to the King Faisal Specialist Hospital & Research Centre, Riyadh, Saudi Arabia, in May 2010 after a sudden-onset headache and loss of consciousness. Brain imaging revealed a large right space-occupying occipital lesion and the patient underwent a craniotomy and resection. Histopathology of the lesion revealed necrotising granulomatous fungal encephalitis with many hyphae engulfed by multinucleated giant histiocytes. Two days later, a computed tomography scan showed debulking of the fungal mass and the patient was discharged on oral voriconazole. However, imaging at a six-week follow-up showed progression of the abnormality. A residual or persistent fungal brain lesion was suspected. Further neurosurgical resection of the lesion was performed and cultures showed growth of Aspergillus flavus. The patient was treated successfully with antifungal therapy over the following two years. PMID:27226920

  9. Genomics of peanut-Aspergillus flavus interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination caused by Aspergillus fungi is a great concern in peanut production worldwide. Pre-harvest Aspergillii infection and aflatoxin contamination are usually severe in peanuts that are grown under drought stressed conditions. Genomic research can provide new tools and resources to...

  10. Transcriptomic profiling of Aspergillus flavus in response to 5-azacytidine.

    PubMed

    Lin, Jian-Qing; Zhao, Xi-Xi; Zhi, Qing-Qing; Zhao, Ming; He, Zhu-Mei

    2013-07-01

    Aspergillus flavus is a common saprophyte and opportunistic pathogen producing aflatoxin (AF) and many other secondary metabolites. 5-Azacytidine (5-AC), a derivative of the nucleoside cytidine, is widely used for studies in epigenetics and cancer biology as an inactivator of DNA methyltransferase and is also used for studying secondary metabolism in fungi. Our previous studies showed that 5-AC affects development and inhibits AF production in A. flavus, and that A. flavus lacks DNA methylation. In this study, an RNA-Seq approach was applied to explore the mechanism of 5-AC's effect on A. flavus. We identified 240 significantly differentially expressed (Q-value<0.05) genes after 5-AC treatment, including two backbone genes respectively in secondary metabolite clusters #27 and #35. These two clusters are involved in development or survival of sclerotia. GO functional enrichment analysis showed that these significantly differentially expressed genes were mainly involved in catalytic activity and proteolytic functions. The expressed transcripts of most genes in the AF biosynthetic gene cluster in A. flavus showed no significant changes after treatment with 5-AC and were expressed at low levels, and the transcription regulator genes aflR and aflS in this cluster did not show differential expression relative to the sample without 5-AC treatment. We found that the veA gene, which encodes protein bridges VelB and LaeA, decreased profoundly the expressed transcripts, and brlA, which encodes an early regulator of development, increased its transcripts in A. flavus after 5-AC treatment. Our data support a model whereby 5-AC affects development through increasing the expression of brlA by depressing the expression of veA and AF production through suppressing veA expression and dysregulating carboxypeptidase activity, which then prevents the aflatoxisomes (vesicles) from performing their normal function in AF formation. Furthermore, the suppressed veA expression weakens or

  11. Genetic variability of Aspergillus flavus isolates from a Mississippi corn field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungus Aspergillus flavus represents a major threat to food safety and food security on a worldwide scale. Corn, peanuts, cotton, rice and edible nuts, can be colonized by A. flavus strains that produce carcinogenic aflatoxins. A biological strategy for control of toxigenic A. flavus starins inv...

  12. Atoxigenic Aspergillus flavus endemic to Italy for biocontrol of aflatoxins in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Effective biological control of aflatoxin­producing Aspergillus flavus with atoxigenic members of that species requires suitable A. flavus well adapted to and resident in target agroecosystems. Eighteen atoxigenic isolates of A. flavus endemic in Italy were compared for ability to reduce aflatoxin c...

  13. Host Genes Involved in the Interaction between Aspergillus flavus and Maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination caused by Aspergillus flavus is a major concern in maize production. Understanding the complex interrelationships of genes during the A. flavus-maize interaction may be the key to developing strategies to interrupt the aflatoxin contamination process. The A. flavus Genome Seq...

  14. Selection of Aspergillus flavus isolates for biological control of aflatoxins in corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungus Aspergillus flavus is responsible for producing carcinogenic mycotoxins, the aflatoxins, on corn (maize) and other crops. An additional harmful toxin, cyclopiazonic acid, is produced by some isolates of A. flavus. Several A. flavus strains that do not produce one or both of these mycoto...

  15. Aspergillus flavus and aflatoxin B1 in flour production.

    PubMed

    Halt, M

    1994-10-01

    This paper discusses the results of investigations of contamination with aflatoxin-producing fungi and aflatoxin B1 affecting 545 samples of wheat grains, 475 samples of intermediate products of wheat grain being milled to flour (like middlings) and 238 samples of flour. A significant contamination with moulds was detected in analyzed samples. Although Aspergillus (34.87%) and Penicillium (32.37%) dominated, other types were also present, e.g., Cladosporium, Fusarium, Mucor, Alternaria, Rhizopus, Absidia and Trichoderma (listed in order of frequency). The presence of Aspergillus flavus, the known aflatoxin producer, was detected in 9.94% of analyzed samples. Isolates of A. Flavus were capable of producing aflatoxin B1 under favourable conditions. Aflatoxin B1 was found in 76.8% of samples contaminated with A. flavus. The highest contamination with aflatoxin B1 was detected in wheat grain samples (mean value of 16.3 micrograms/kg) and in intermediate products of wheat grain being milled to flour (mean value of 11.13 micrograms/kg). Contamination was lower in flour samples (mean value of 4.13 micrograms/kg). With regard to proposed standards given by the FAO and WHO, under which the content of aflatoxin should not exceed 30 micrograms/kg in food products, only two of 96 samples did not meet these criteria. PMID:7859854

  16. Effect of gamma radiation on Aspergillus flavus and Aspergillus ochraceus ultrastructure and mycotoxin production

    NASA Astrophysics Data System (ADS)

    Ribeiro, J.; Cavaglieri, L.; Vital, H.; Cristofolini, A.; Merkis, C.; Astoreca, A.; Orlando, J.; Carú, M.; Dalcero, A.; Rosa, C. A. R.

    2011-05-01

    The aim of this work was to study the effect of gamma radiation (2 kGy) on Aspergillus flavus and Aspergillus ochraceus ultrastructure. Moreover, the influence on aflatoxin B 1 and ochratoxin A production was also observed. Irradiated A. flavus strain showed a dull orangish colony while control strain showed the typical green color. Minor differences were observed on stipes, metulae and conidia size between control and irradiated A. flavus and A. ochraceus strains. Irradiated fungi showed ultrastructural changes on cell wall, plasmalema and cytoplasm levels. The levels of mycotoxins produced by irradiated strains were two times greater than those produced by control strains. Successive transferences of irradiated strains on malt extract agar allowed the fungus to recuperate morphological characteristics. Although minor changes in the fungal morphology were observed, ultrastructural changes at cell wall level and the increase of mycotoxin production ability were observed. Inappropriate storage of irradiated food and feed would allow the development of potentially more toxicogenic fungal propagules.

  17. The population genomics of mycotoxin diversity in Aspergillus flavus and Aspergillus parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycotoxins, and especially the aflatoxins, are an enormous problem in agriculture, with aflatoxin B1 being the most carcinogenic known natural compound. The worldwide costs associated with aflatoxin monitoring and crop losses are in the hundreds of millions of dollars. Aspergillus flavus and A. par...

  18. Development and evaluation of peanut germplasm with resistance to Aspergillus flavus from core collection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanut (Arachis hypogaea L.), one of the main oil and cash crops in the world, is susceptible to Aspergillus flavus, resulting in loss in quality. Aspergillus flavus infection is a problem for peanut production and industry in China. Therefore, it is imperative to develop new peanut germplasm with ...

  19. Structure analysis of an Aspergillus flavus kernels population in North Italy. First analysis of an Aspergillus flavus kernels population based on vegetative compatibility groups in Northern Italy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In order to gain insight into the causal agents of aflatoxin contamination of maize in Italy, populations of Aspergillus flavus on maize produced in the most affected area were characterized. Forty-six percent of A. flavus, isolated from maize kernels collected in 5 districts of northern Italy betwe...

  20. Investigations on the Antifungal Effect of Nerol against Aspergillus flavus Causing Food Spoilage

    PubMed Central

    Tian, Jun; Zeng, Xiaobin; Zeng, Hong; Feng, Zhaozhong; Miao, Xiangmin; Peng, Xue

    2013-01-01

    The antifungal efficacy of nerol (NEL) has been proved against Aspergillus flavus by using in vitro and in vivo tests. The mycelial growth of A. flavus was completely inhibited at concentrations of 0.8 μL/mL and 0.1 μL/mL NEL in the air at contact and vapor conditions, respectively. The NEL also had an evident inhibitory effect on spore germination in A. flavus along with NEL concentration as well as time-dependent kinetic inhibition. The NEL presented noticeable inhibition on dry mycelium weight and synthesis of aflatoxin B1 (AFB1) by A. flavus, totally restraining AFB1 production at 0.6 μL/mL. In real food system, the efficacy of the NEL on resistance to decay development in cherry tomatoes was investigated in vivo by exposing inoculated and control fruit groups to NEL vapor at different concentration. NEL vapors at 0.1 μL/mL air concentration significantly reduced artificially contaminated A. flavus and a broad spectrum of fungal microbiota. Results obtained from presented study showed that the NEL had a great antifungal activity and could be considered as a benefit and safe tool to control food spoilage. PMID:24453813

  1. Loss of msnA, a Putative Stress Regulatory Gene, in Aspergillus parasiticus and Aspergillus flavus Increased Production of Conidia, Aflatoxins and Kojic Acid

    PubMed Central

    Chang, Perng-Kuang; Scharfenstein, Leslie L.; Luo, Meng; Mahoney, Noreen; Molyneux, Russell J.; Yu, Jiujiang; Brown, Robert L.; Campbell, Bruce C.

    2011-01-01

    Production of the harmful carcinogenic aflatoxins by Aspergillus parasiticus and Aspergillus flavus has been postulated to be a mechanism to relieve oxidative stress. The msnA gene of A. parasiticus and A. flavus is the ortholog of Saccharomyces cerevisiae MSN2 that is associated with multi-stress response. Compared to wild type strains, the msnA deletion (∆msnA) strains of A. parasiticus and A. flavus exhibited retarded colony growth with increased conidiation. The ∆msnA strains also produced slightly higher amounts of aflatoxins and elevated amounts of kojic acid on mixed cereal medium. Microarray assays showed that expression of genes encoding oxidative stress defense enzymes, i.e., superoxide dismutase, catalase, and cytochrome c peroxidase in A. parasiticus ∆msnA, and the catalase A gene in A. flavus ∆msnA, was up-regulated. Both A. parasiticus and A. flavus ∆msnA strains produced higher levels of reactive oxygen species (ROS), and ROS production of A. flavus msnA addback strains was decreased to levels comparable to that of the wild type A. flavus. The msnA gene appears to be required for the maintenance of the normal oxidative state. The impairment of msnA resulted in the aforementioned changes, which might be used to combat the increased oxidative stress in the cells. PMID:22069691

  2. How Peroxisomes Affect Aflatoxin Biosynthesis in Aspergillus Flavus

    PubMed Central

    Reverberi, Massimo; Punelli, Marta; Smith, Carrie A.; Zjalic, Slaven; Scarpari, Marzia; Scala, Valeria; Cardinali, Giorgia; Aspite, Nicaela; Pinzari, Flavia; Payne, Gary A.; Fabbri, Anna A.; Fanelli, Corrado

    2012-01-01

    In filamentous fungi, peroxisomes are crucial for the primary metabolism and play a pivotal role in the formation of some secondary metabolites. Further, peroxisomes are important site for fatty acids β-oxidation, the formation of reactive oxygen species and for their scavenging through a complex of antioxidant activities. Oxidative stress is involved in different metabolic events in all organisms and it occurs during oxidative processes within the cell, including peroxisomal β-oxidation of fatty acids. In Aspergillus flavus, an unbalance towards an hyper-oxidant status into the cell is a prerequisite for the onset of aflatoxin biosynthesis. In our preliminary results, the use of bezafibrate, inducer of both peroxisomal β-oxidation and peroxisome proliferation in mammals, significantly enhanced the expression of pex11 and foxA and stimulated aflatoxin synthesis in A. flavus. This suggests the existence of a correlation among peroxisome proliferation, fatty acids β-oxidation and aflatoxin biosynthesis. To investigate this correlation, A. flavus was transformed with a vector containing P33, a gene from Cymbidium ringspot virus able to induce peroxisome proliferation, under the control of the promoter of the Cu,Zn-sod gene of A. flavus. This transcriptional control closely relates the onset of the antioxidant response to ROS increase, with the proliferation of peroxisomes in A. flavus. The AfP33 transformant strain show an up-regulation of lipid metabolism and an higher content of both intracellular ROS and some oxylipins. The combined presence of a higher amount of substrates (fatty acids-derived), an hyper-oxidant cell environment and of hormone-like signals (oxylipins) enhances the synthesis of aflatoxins in the AfP33 strain. The results obtained demonstrated a close link between peroxisome metabolism and aflatoxin synthesis. PMID:23094106

  3. Inhibition of Aspergillus flavus in soil by antagonistic Pseudomonas strains reduces the potential for airborne spore dispersal

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pseudomonas chlororaphis strain JP1015 and Pseudomonas fluorescens strain JP2175 were previously isolated from Mississippi cornfield soil samples and selected for their growth inhibition of Aspergillus flavus in laboratory culture. In this study, the antifungal activity of these bacterial strains a...

  4. Development of a GFP-Expressing Aspergillus flavus Strain to Study Fungal Invasion, Colonization, and Resistance in Cottonseed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cotton bolls were inoculated with a green fluorescent protein (GFP)-expressing Aspergillus flavus (strain 70) to monitor fungal growth, mode of entry, colonization of cottonseeds and production of aflatoxins. The GFP strain and the wild type did not differ significantly in pathogen aggressiveness a...

  5. Effects of Nutrients in Substrates of Different Grains on Aflatoxin B1 Production by Aspergillus flavus

    PubMed Central

    Liu, Jie; Sun, Lvhui; Zhang, Niya; Zhang, Jiacai; Guo, Jiao; Li, Chong; Rajput, Shahid Ali; Qi, Desheng

    2016-01-01

    The current study was to better understand the potential factors affecting aflatoxin B1 (AFB1) accumulation varies between different grains. The nutrient composition and contents of defatted substrates were determined; additionally, according to the nutrient content of the substrates, the effects of starch, soluble sugars, amino acids, and trace elements on AFB1 production and mycelial growth in Czapek-Dox medium were examined. These results verified that removal of lipids from ground substrates significantly reduced the substrate's potential for AFB1 production by Aspergillus flavus. Maltose, glucose, sucrose, arginine, glutamic acid, aspartic acid, and zinc significantly induced AFB1 production up to 1.7- to 26.6-fold. And stachyose more significantly promoted A. flavus growth than the other nutrients. Thus, this study demonstrated that, combined with the nutrients content of grains, in addition to lipids, sucrose, stachyose, glutamic acid, and zinc might play key roles in various grains that are differentially infected by A. flavus. Particularly, two new nutrients (arginine and stachyose) of the grains we found significantly stimulate AFB1 production and A. flavus growth, respectively. The results provide new concepts for antifungal methods to protect food and animal feed from AFB1 contamination. PMID:27294129

  6. Effects of Nutrients in Substrates of Different Grains on Aflatoxin B1 Production by Aspergillus flavus.

    PubMed

    Liu, Jie; Sun, Lvhui; Zhang, Niya; Zhang, Jiacai; Guo, Jiao; Li, Chong; Rajput, Shahid Ali; Qi, Desheng

    2016-01-01

    The current study was to better understand the potential factors affecting aflatoxin B1 (AFB1) accumulation varies between different grains. The nutrient composition and contents of defatted substrates were determined; additionally, according to the nutrient content of the substrates, the effects of starch, soluble sugars, amino acids, and trace elements on AFB1 production and mycelial growth in Czapek-Dox medium were examined. These results verified that removal of lipids from ground substrates significantly reduced the substrate's potential for AFB1 production by Aspergillus flavus. Maltose, glucose, sucrose, arginine, glutamic acid, aspartic acid, and zinc significantly induced AFB1 production up to 1.7- to 26.6-fold. And stachyose more significantly promoted A. flavus growth than the other nutrients. Thus, this study demonstrated that, combined with the nutrients content of grains, in addition to lipids, sucrose, stachyose, glutamic acid, and zinc might play key roles in various grains that are differentially infected by A. flavus. Particularly, two new nutrients (arginine and stachyose) of the grains we found significantly stimulate AFB1 production and A. flavus growth, respectively. The results provide new concepts for antifungal methods to protect food and animal feed from AFB1 contamination. PMID:27294129

  7. Copper induction and differential expression of laccase in Aspergillus flavus.

    PubMed

    Gomaa, Ola M; Momtaz, Osama A

    2015-03-01

    Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus . Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent. PMID:26221119

  8. Role of 3-Nitropropanoic Acid in Nitrate Formation by Aspergillus flavus1

    PubMed Central

    Doxtader, K. G.; Alexander, M.

    1966-01-01

    Doxtader, K. G. (Cornell University, Ithaca, N.Y.), and M. Alexander. Role of 3-nitropropanoic acid in nitrate formation by Aspergillus flavus. J. Bacteriol. 91:1186–1191. 1966.—Aspergillus flavus formed nitrate, 3-nitropropanoic acid (3-NPA), kojic acid, and a substance tentatively identified as N-formyl-N-hydroxy-glycine during growth in a medium with ammonium as sole nitrogen source. The concentration of the nitro compound reached a maximum prior to the appearance of nitrate; the 3-NPA level subsequently decreased with a concomitant increase in nitrate concentration. Replacement cultures of A. flavus produced nitrate from culture filtrates containing 3-NPA or from synthetic 3-NPA but not when supplied with fresh ammonium-sucrose medium, the nitrate-nitrogen formed being equivalent to 50% of the quantity of the 3-NPA-nitrogen initially present. Neither nitrate nor 3-NPA was synthesized by the fungus during growth in media with low pH or low ammonium concentrations. It is proposed that 3-NPA is either an intermediate or is in equilibrium with an intermediate in nitrification by the fungus. PMID:5929750

  9. Comparative Chemistry of Aspergillus oryzae (RIB40) and A. flavus (NRRL 3357)

    PubMed Central

    Rank, Christian; Klejnstrup, Marie Louise; Petersen, Lene Maj; Kildgaard, Sara; Frisvad, Jens Christian; Gotfredsen, Charlotte Held; Larsen, Thomas Ostenfeld

    2012-01-01

    Aspergillus oryzae and A. flavus are important species in industrial biotechnology and food safety and have been some of the first aspergilli to be fully genome sequenced. Bioinformatic analysis has revealed 99.5% gene homology between the two species pointing towards a large coherence in the secondary metabolite production. In this study we report on the first comparison of secondary metabolite production between the full genome sequenced strains of A. oryzae (RIB40) and A. flavus (NRRL 3357). Surprisingly, the overall chemical profiles of the two strains were mostly very different across 15 growth conditions. Contrary to previous studies we found the aflatrem precursor 13-desoxypaxilline to be a major metabolite from A. oryzae under certain growth conditions. For the first time, we additionally report A. oryzae to produce parasiticolide A and two new analogues hereof, along with four new alkaloids related to the A. flavus metabolites ditryptophenalines and miyakamides. Generally the secondary metabolite capability of A. oryzae presents several novel end products likely to result from the domestication process from A. flavus. PMID:24957367

  10. Use of Pyrosequencing to Quantify Incidence of a Specific Aspergillus flavus Strain Within Complex Fungal Communities Associated with Commercial Cotton Crops

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are highly toxic carcinogens produced by several species of Aspergillus, and its presence in foods causes chronic health effects including immune-system suppression, growth retardation, cancer, and death in both humans and domestic animals. Atoxigenic strains of Aspergillus flavus have b...

  11. AMINO ACID SUPPLEMENTATION REVEALS DIFFERENTIAL REGULATION OF AFLATOXIN BIOSYNTHESIS IN ASPERGILLUS FLAVUS NRRL 3357 AND ASPERGILLUS PARASITICUS SRRC 143

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic and carcinogenic secondary metabolites produced by the fungi Aspergillus flavus and A. parasiticus. In order to better understand the molecular mechanisms that regulate aflatoxin production, the biosynthesis of the toxin in A. flavus and A. parasiticus grown in yeast extract su...

  12. Antibiotic Extraction as a Recent Biocontrol Method for Aspergillus Niger andAspergillus Flavus Fungi in Ancient Egyptian mural paintings

    NASA Astrophysics Data System (ADS)

    Hemdan, R.; et al.

    Biodeterioration of mural paintings by Aspergillus niger and Aspergillus flavus Fungi has been proved in different mural paintings in Egypt nowadays. Several researches have studied the effect of fungi on mural paintings, the mechanism of interaction and methods of control. But none of these researches gives us the solution without causing a side effect. In this paper, for the first time, a recent treatment by antibiotic "6 penthyl α pyrone phenol" was applied as a successful technique for elimination of Aspergillus niger and Aspergillus flavus. On the other hand, it is favorable for cleaning Surfaces of Murals executed by tembera technique from the fungi metabolism which caused a black pigments on surfaces.

  13. Evaluation of intraspecific competition (Aspergillus flavus Link) and aflatoxin formation in suspended disc culture and preharvest maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The abilities of non-aflatoxin producing strains of Aspergillus flavus NRRL 32354; 18543; 21882; 21368 as well as domesticated koji strains Aspergillus oryzae (syn. A. flavus var. oryzae) NRRL 451; 1911; 5592; 6271; 30038 to interfere with aflatoxin formation by A. flavus NRRL 3357; 32355 were exami...

  14. The Aspergillus flavus Histone Acetyltransferase AflGcnE Regulates Morphogenesis, Aflatoxin Biosynthesis, and Pathogenicity

    PubMed Central

    Lan, Huahui; Sun, Ruilin; Fan, Kun; Yang, Kunlong; Zhang, Feng; Nie, Xin Y.; Wang, Xiunai; Zhuang, Zhenhong; Wang, Shihua

    2016-01-01

    Histone acetyltransferases (HATs) help regulate fungal development and the production of secondary metabolites. In this study, we determined that the HAT AflGcnE influenced morphogenesis and aflatoxin biosynthesis in Aspergillus flavus. We observed that AflGcnE localized to the nucleus and cytoplasm during the conidial production and germination stages, while it was located mainly in the nucleus during the hyphal development stage. Deletion of AflgcnE inhibited the growth of A. flavus and decreased the hydrophobicity of the cell surface. The ΔAflgcnE mutant exhibited a lack of asexual sporulation and was unable to generate sclerotia. Additionally, AflgcnE was required to maintain cell wall integrity and genotoxic stress responses. Importantly, the ΔAflgcnE mutant did not produce aflatoxins, which was consistent with a significant down-regulation of aflatoxin gene expression levels. Furthermore, our data revealed that AflgcnE is a pathogenicity factor required for colonizing maize seeds. In summary, we revealed that A. flavus AflGcnE is crucial for morphological development, aflatoxin biosynthesis, stress responses, and pathogenicity. Our findings help clarify the functional divergence of GcnE orthologs, and may provide a possible target for controlling A. flavus infections of agriculturally important crops. PMID:27625637

  15. The Aspergillus flavus Histone Acetyltransferase AflGcnE Regulates Morphogenesis, Aflatoxin Biosynthesis, and Pathogenicity.

    PubMed

    Lan, Huahui; Sun, Ruilin; Fan, Kun; Yang, Kunlong; Zhang, Feng; Nie, Xin Y; Wang, Xiunai; Zhuang, Zhenhong; Wang, Shihua

    2016-01-01

    Histone acetyltransferases (HATs) help regulate fungal development and the production of secondary metabolites. In this study, we determined that the HAT AflGcnE influenced morphogenesis and aflatoxin biosynthesis in Aspergillus flavus. We observed that AflGcnE localized to the nucleus and cytoplasm during the conidial production and germination stages, while it was located mainly in the nucleus during the hyphal development stage. Deletion of AflgcnE inhibited the growth of A. flavus and decreased the hydrophobicity of the cell surface. The ΔAflgcnE mutant exhibited a lack of asexual sporulation and was unable to generate sclerotia. Additionally, AflgcnE was required to maintain cell wall integrity and genotoxic stress responses. Importantly, the ΔAflgcnE mutant did not produce aflatoxins, which was consistent with a significant down-regulation of aflatoxin gene expression levels. Furthermore, our data revealed that AflgcnE is a pathogenicity factor required for colonizing maize seeds. In summary, we revealed that A. flavus AflGcnE is crucial for morphological development, aflatoxin biosynthesis, stress responses, and pathogenicity. Our findings help clarify the functional divergence of GcnE orthologs, and may provide a possible target for controlling A. flavus infections of agriculturally important crops. PMID:27625637

  16. RNA-Seq-Based Transcriptome Analysis of Aflatoxigenic Aspergillus flavus in Response to Water Activity

    PubMed Central

    Zhang, Feng; Guo, Zhenni; Zhong, Hong; Wang, Sen; Yang, Weiqiang; Liu, Yongfeng; Wang, Shihua

    2014-01-01

    Aspergillus flavus is one of the most important producers of carcinogenic aflatoxins in crops, and the effect of water activity (aw) on growth and aflatoxin production of A. flavus has been previously studied. Here we found the strains under 0.93 aw exhibited decreased conidiation and aflatoxin biosynthesis compared to that under 0.99 aw. When RNA-Seq was used to delineate gene expression profile under different water activities, 23,320 non-redundant unigenes, with an average length of 1297 bp, were yielded. By database comparisons, 19,838 unigenes were matched well (e-value < 10−5) with known gene sequences, and another 6767 novel unigenes were obtained by comparison to the current genome annotation of A. flavus. Based on the RPKM equation, 5362 differentially expressed unigenes (with |log2Ratio| ≥ 1) were identified between 0.99 aw and 0.93 aw treatments, including 3156 up-regulated and 2206 down-regulated unigenes, suggesting that A. flavus underwent an extensive transcriptome response during water activity variation. Furthermore, we found that the expression of 16 aflatoxin producing-related genes decreased obviously when water activity decreased, and the expression of 11 development-related genes increased after 0.99 aw treatment. Our data corroborate a model where water activity affects aflatoxin biosynthesis through increasing the expression of aflatoxin producing-related genes and regulating development-related genes. PMID:25421810

  17. Potential of essential oils for protection of grains contaminated by aflatoxin produced by Aspergillus flavus

    PubMed Central

    Esper, Renata H.; Gonçalez, Edlayne; Marques, Marcia O. M.; Felicio, Roberto C.; Felicio, Joana D.

    2014-01-01

    Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic metabolite produced by Aspergillus species on food and agricultural commodities. Inhibitory effects of essential oils of Ageratum conyzoides (mentrasto) and Origanum vulgare (oregano) on the mycelial growth and aflatoxin B1 production by Aspergillus flavus have been studied previously in culture medium. The aim of this study was to evaluate aflatoxin B1 production by Aspergillus flavus in real food systems (corn and soybean) treated with Ageratum conyzoides (mentrasto) and Origanum vulgare (oregano) essential oils. Samples with 60 g of the grains were treated with different volumes of essential oils, 200, 100, 50, and 10 μL for oregano and 50, 30, 15, and 10 μL for mentrasto. Fungal growth was evaluated by disk diffusion method. Aflatoxin B1 production was evaluated inoculating suspensions of A. flavus containing 1.3 × 105 spores/mL in 60 g of grains (corn and soybeans) after adjusting the water activity at 0.94. Aflatoxin was quantified by photodensitometry. Fungal growth and aflatoxin production were inhibited by essential oils, but the mentrasto oil was more effective in soybeans than that of oregano. On the other hand, in corn samples, the oregano essential oil was more effective than that of mentrasto. Chemical compositions of the essential oils were also investigated. The GC/MS oils analysis showed that the main component of mentrasto essential oil is precocene I and of the main component of oregano essential oil is 4-terpineol. The results indicate that both essential oils can become an alternative for the control of aflatoxins in corn and soybeans. PMID:24926289

  18. Dual genome microarray: Fusarium verticillioides and Aspergillus flavus gene expression in co-culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins produced by Aspergillus flavus, and fumonisins produced by Fusarium verticillioides, are prominent among the mycotoxins associated with economic losses to the maize grain industry worldwide. F. verticillioides is also recognized as a systemic endophyte of maize that prevents opportunisti...

  19. Twenty-four microsatellite markers for the aflatoxin-producing fungus Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus infects both plants and humans and contaminates diverse agricultural crops with aflatoxins, highly carcinogenic fungal metabolites. We describe 24 microsatellite markers developed to assess genetic diversity and recombination within and between three vegetative compatibility group...

  20. Gene Profiling for Studying the Mechanism of Aflatoxin Biosynthesis in Aspergillus flavus and A. parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic and carcinogenic polyketide metabolites produced by certain fungal species, including Aspergillus flavus and A. parasiticus. Because many internal and external factors, such as nutrition and environment affect aflatoxin biosynthesis, we have analyzed the transcriptome of A. fla...

  1. Evaluation of Atoxigenic Strains of Aspergillus flavus as Potential Biocontrol Agents for Aflatoxin in Maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination resulting from maize infection by Aspergillus flavus is both an economic concern and public health concern. Therefore, strategies for controlling maize contamination are being investigated. Abilities of 11 naturally occurring atoxigenic strains in Nigeria to reduce aflatox...

  2. Recombination, balancing selection and geographic subdivision among worldwide populations of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a global agent of aflatoxin contamination of economically important crops such as corn, peanuts, and cottonseed. Extensive studies have elucidated the biochemical and regulatory mechanisms of aflatoxin production, but basic knowledge of the evolutionary processes that maintain ...

  3. Flocculation behavior and mechanism of bioflocculant produced by Aspergillus flavus.

    PubMed

    Aljuboori, Ahmad H Rajab; Idris, Azni; Al-joubory, Hamid Hussain Rijab; Uemura, Yoshimitsu; Ibn Abubakar, B S U

    2015-03-01

    In this study, the flocculation behavior and mechanism of a cation-independent bioflocculant IH-7 produced by Aspergillus flavus were investigated. Results showed 91.6% was the lowest flocculating rate recorded by IH-7 (0.5 mg L(-1)) at pH range 4-8. Moreover, IH-7 showed better flocculation performance than polyaluminum chloride (PAC) at a wide range of flocculant concentration (0.06-25 mg L(-1)), temperature (5-45 °C) and salinity (10-60% w/w). The current study found that cation addition did not significantly enhance the flocculating rate and IH-7 is a positively charged bioflocculant. These findings suggest that charge neutralization is the main flocculation mechanism of IH-7 bioflocculant. IH-7 was significantly used to flocculate different types of suspended solids such as activated carbons, kaolin clays, soil solids and yeast cells. PMID:25560664

  4. Effect of sexual recombination on population diversity in aflatoxin production by Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is the major producer of carcinogenic aflatoxins (AFs) in crops worldwide. Recent efforts to reduce AF concentrations in crops have focused on the use of two non-aflatoxigenic A. flavus strains, AF36 and NRRL 21882 (Afla-Guard), as biological control agents. These products are a...

  5. A GOOD ENDOPHYTE OF MAIZE: ACREMONIUM ZEAE ANTIBIOTICS INHIBITORY TO ASPERGILLUS FLAVUS AND FUSARIUM VERTICILLIOIDES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The maize endophyte Acremonium zeae Gams and Sumner is antagonistic to kernel rotting and mycotoxin producing fungi Aspergillus flavus and Fusarium verticillioides in cultural tests for antagonism and interferes with A. flavus infection and aflatoxin contamination of preharvest maize kernels. Chemi...

  6. EST Profiling for Elucidation of Molecular Regulation of Aflatoxin bBiosynthesis in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic and carcinogenic polyketide metabolites produced by fungal species, including Aspergillus flavus and A. parasiticus. Many internal and external factors, such as nutrition and environment, affect aflatoxin biosynthesis. A. flavus EST has been carried out and a microarray has be...

  7. Aspergillus Flavus/Aflatoxin Occurrence and Expression of Aflatoxin Biosynthesis Genes in Soil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins (AF) are carcinogenic metabolites produced by several species of Aspergillus, including A. flavus. Although A. flavus is readily isolated from environmental samples, soil and plant material are considered the natural habitat of this fungus. Studies were conducted on a Dundee silt loam to ...

  8. Volatile profiles of toxigenic and non-toxigenic Aspergillus flavus using SPME for solid phase extraction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Toxigenic and atoxigenic strains of Aspergillus flavus were grown on potato dextrose agar (PDA) and wetted sterile, cracked corn for 21 and 14 days, respectively. Volatile compounds produced by A. flavus, as well as those present in the PDA controls and sterile cracked corn, were collected using sol...

  9. Formation of Aspergillus flavus sclerotia on corn grown under different drought stress conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a major producer of carcinogenic aflatoxins worldwide in corn, peanuts, tree nuts, cottonseed, spices and other crops. Many countries have strict limits on the amount of aflatoxins permitted in human commodities and animal feed. Sclerotia produced by A. flavus serve several f...

  10. Green Fluorescent Protein as a Reporter To Monitor Gene Expression and Food Colonization by Aspergillus flavus

    PubMed Central

    Du, Wanglei; Huang, Zhengyu; Flaherty, Joseph E.; Wells, Kevin; Payne, Gary A.

    1999-01-01

    Transformants of Aspergillus flavus containing the Aequorea victoria gfp gene fused to a viral promoter or the promoter region and 483 bp of the coding region of A. flavus aflR expressed green fluorescence detectable without a microscope or filters. Expression of green fluorescent protein fluorescence was correlated with resistance to aflatoxin accumulation in five corn genotypes inoculated with these transformants. PMID:9925624

  11. The release of the Aspergillus flavus whole genome sequence for public access

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are produced by Aspergillus flavus and A. parasiticus. These toxic and carcinogenic compounds contaminate pre-harvest agricultural crops in the field and post-harvest grains during storage. In order to reduce and eliminate aflatoxin contamination of food and feed, Aspergilllus flavus wh...

  12. Integrated database for identifying candate genes for Aspergillus flavus resistance in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus Link:Fr, an opportunistic fungus that produces aflatoxin, is pathogenic to maize and other oilseed crops. Aflatoxin is a potent carcinogen, and its presence markedly reduces the value of grain. Understanding and enhancing host resistance to A. flavus infection and/or subsequent af...

  13. IDENTIFICATION OF MAIZE KERNEL ENDOSPERM PROTEINS ASSOCIATED WITH RESISTANCE TO AFLATOXIN CONTAMINATION BY ASPERGILLUS FLAVUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are carcinogens produced mainly by Aspergillus flavus during infection of susceptible crops, such as maize (Zea mays L.). Previously, embryo proteins from maize genotypes resistant or susceptible to A. flavus infection were compared using proteomics and resistance-associated proteins wer...

  14. Recombination and lineage-specific gene loss in the aflatoxin gene cluster of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins produced by Aspergillus flavus are potent carcinogens that contaminate agricultural crops. Recent efforts to reduce aflatoxin concentrations in crops have focused on biological control using nonaflatoxigenic A. flavus strains AF36 (= NRRL 18543) and NRRL 21882 (the active component of af...

  15. Developing an in vitro method to assess aflatoxin biosynthesis suppression in Aspergillus flavus through RNAi technologies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The soil-inhabitant fungus Aspergillus flavus is consistently associated with agronomical fields, where it promptly colonizes important crops such as corn (Zea mays) and peanuts (Arachis hypogaea). The consumption of A. flavus-contaminated of food grains poses a potential threat for human and animal...

  16. The launch of the Aspergillus flavus genome browser and limited release of whole genome sequence

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are produced by Aspergillus flavus and A. parasiticus. These toxic and carcinogenic compounds contaminate pre-harvest agricultural crops in the field and post-harvest grains during storage. In order to reduce and eliminate aflatoxin contamination of food and feed, Aspergilllus flavus wh...

  17. Characterization of an Aspergillus flavus alkaline protease and its role in the infection of maize kernels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A 33 kDa protein present in Aspergillus flavus infected maize embryo tissue was identified as a fungal alkaline protease (ALP). This protein became one of the major extracellular proteins of A. flavus in potato dextrose broth medium cultural filtrate after 3 days, but was expressed at low levels or ...

  18. Aspergillus flavus Genomic Data Mining Provides Clues for Its Use in Producing Biobased Products

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is notorious for its ability to produce aflatoxins. It is also an opportunistic pathogen that infects plants, animals and human beings. The ability to survive in the natural environment, living on plant tissues (leaves or stalks), live or dead insects make A. flavus a ubiquitous...

  19. A two-dimenstional proteome reference map of the aflatoxigenic fungus Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The description of A. flavus proteome provides insight into its basic biology and a basis for its future proteomic investigations. Aspergillus flavus is a widely distributed fungal pathogen that infects important agricultural commodities (maize, tree nuts, etc.) and contaminates them with aflatoxin...

  20. Molecular markers associated with resistance to Aspergillus flavus in maize: QTL and discriminant analyses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination of maize (Zea mays L.) grain caused by Aspergillus flavus is a serious health hazard to animals and humans. Resistance to infection by A. flavus is poorly understood. The objectives of this investigation were to identify potential candidate markers associated with resistance ...

  1. Genetic Isolation among Sympatric Vegetative Compatibility Groups of the aflatoxin-producing fungus Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus, fungal pathogen of animals and both wild and economically important plants, is most recognized for producing aflatoxin, a cancer-causing secondary metabolite, that contaminates food and animal feed globally. A. flavus is asexual and has a vegetative incompatibility system that li...

  2. Sexual recombination and the possibility of cryptic heterokaryosis in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus infects both plants and animals and is of toxicological importance due to its production of aflatoxins (AFs). Recent efforts to reduce AF concentrations have focused on the use of the biocontrols AF36 and Afla-Guard®, both of which contain nonaflatoxigenic A. flavus strains as an ...

  3. Sexual reproduction in Aspergillus flavus sclerotia: acquisition of novel alleles from soil populations and uniparental mitochondrial inheritance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus colonizes agricultural commodities worldwide and contaminates them with carcinogenic aflatoxins. The high genetic diversity of A. flavus populations is largely due to sexual reproduction characterized by the formation of ascospore-bearing ascocarps embedded within sclerotia. A. ...

  4. [Aflatoxins produced by Aspergillus flavus in soya and other legumes].

    PubMed

    Topsy, K

    1977-01-01

    There is no doubt that our programme of applied nutrition must include soya on account of the high nutritive value of the legume. This underlines research undertaken here regarding the risks of contamination by A. flavus and the subsequent formation of aflatoxins on and in soya beans. We have studied on parallel lines soya beans and other legumes important in the local dietary habits. These legumes are either obtained locally or imported. On every specimen of legume we have tried to confirm, or otherwise, the presence of A. flavus and the aflatoxins. This was followed by experimenting on the conditions for growth and formation of aflatoxins on every one of the legumes. During subsequent experiments we have studied mixtures of legumes containing soya. Research on these lines has shown the inhibitory effects of legumes such as lentils, dried garden peas, Bengal gram, green peas, red peas, and broad beans on the growth of A. flavus and the formation of aflatoxins. Groundnut, on the other hand, seems to encourage such growth and such formation. The results of the experiments carried out, as above, lead us to conclude that soya must form part of our national food monitoring programme. PMID:418721

  5. Community structure of Aspergillus flavus and persistence of the atoxigenic strain A flavus AF36 in applied fields

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic and carcinogenic metabolites produced by several fungi in Aspergillus Section Flavi that frequently contaminate crops. Aflatoxins impact the value of crops. The use of atoxigenic strains of A. flavus to displace aflatoxin producers is a proven method to reduce aflatoxin contamin...

  6. Non-aflatoxigenic Aspergillus flavus isolates reduce aflatoxins, cyclopiazonic acid and fumonisin in corn (maize)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus strains vary widely in their production of aflatoxins and cyclopiazonic acid (CPA). A total of 500 Aspergillus strains isolated from a variety of sources showed 16.4% were negative for both aflatoxin and CPA, 41.3% were positive for both mycotoxins, 13.0% were positive only fo...

  7. NsdC and NsdD affect Aspergillus flavus morphogenesis and aflatoxin production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The transcription factors NsdC and NsdD have been shown to be necessary for sexual development in Aspergillus nidulans. Herein we examine the role of these proteins in development and aflatoxin production of the agriculturally important, aflatoxin-producing fungus, Aspergillus flavus. We found tha...

  8. Sexual reproduction influences aflatoxin chemotype diversity in worldwide populations of Aspergillus flavus and A. parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic polyketides produced by several Aspergillus species that contaminate food crops worldwide. Aspergillus flavus and A. parasiticus are the most common agents of aflatoxin contamination of oil-rich crops. The genes involved in aflatoxin biosynthesis are clustered and convert acetat...

  9. rmtA, encoding a putative anginine methyltransferase, regulates secondary metabolism and development in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is found colonizing numerous oil seed crops such as corn, peanuts, sorghum, treenuts and cotton worldwide, contaminating them with aflatoxin and other harmful potent toxins. In the phylogenetically related model fungus Aspergillus nidulans, the methyltransferase, RmtA, has been de...

  10. RNA interference reduces aflatoxin accumulation by Aspergillus flavus in peanut seeds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are among the most powerful carcinogens in nature. They are produced by the fungal pathogen Aspergillus flavus Link and other Aspergillus species. Aflatoxins accumulate in many crops, including rice, wheat, oats, pecans, pistachios, soybean, cassava, almonds, peanuts, beans, corn and cot...

  11. Population genetics as a tool for understanding toxigenesis in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Species in Aspergillus section Flavi commonly infect agricultural staples such as corn, peanuts, cottonseed, and tree nuts and produce an array of mycotoxins, the most potent of which is aflatoxin. Aspergillus flavus is the dominant aflatoxin-producing species in the majority of crops. Populations...

  12. Antifungal properties and inhibitory effects upon aflatoxin production of Thymus vulgaris L. by Aspergillus flavus Link.

    PubMed

    Kohiyama, Cássia Yumie; Yamamoto Ribeiro, Milene Mayumi; Mossini, Simone Aparecida Galerani; Bando, Erika; Bomfim, Natália da Silva; Nerilo, Samuel Botião; Rocha, Gustavo Henrique Oliveira; Grespan, Renata; Mikcha, Jane Martha Graton; Machinski, Miguel

    2015-04-15

    The antifungal and antiaflatoxigenic properties of Thymus vulgaris essential oil (TEO) were evaluated upon Aspergillus flavus "in vitro". Suspension containing 10(6) of A. flavus were cultivated with TEO in concentrations ranging from 50 to 500 μg/mL. TEO reached minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) at 250 μg/mL. Inhibition of ergosterol biosynthesis was detected at a concentration of 100 μg/mL of TEO. Morphological evaluation performed by both light microscopy and scanning electron microscopy showed that antifungal activity of TEO could be detected starting at a concentration of 50 μg/mL and the fungicide effect at a concentration of 250 μg/mL. TEO completely inhibited production of both B1 and B2 aflatoxins (AFB1 and AFB2) at a concentration of 150 μg/mL. This way, fungal biomass development and aflatoxin production were dependent on TEO concentration. Therefore, TEO was capable of controlling the growth of A. flavus and its production of aflatoxins. PMID:25466118

  13. Lack of aflatoxin production by Aspergillus flavus on a resistant peanut line is associated with delayed expression of aflatoxin genes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins, produced by Aspergillus flavus and Aspergillus parasiticus, are the most toxic fungal secondary metabolites and the most potent carcinogens that contaminate agricultural commodities such as peanuts, cotton and corn. Understanding the underlying mechanisms of crop resistance to fungal in...

  14. Development and Evaluation of an Affymetrix array for Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A multi-species Affymetrix GeneChip array was developed to study development, metabolism and pathogenicity of A. flavus. This chip based on the whole genome sequence of A. flavus, contains 13,000 A. flavus genes, 8,000 maize genes and 25 human and mouse innate immune response genes, as well as the ...

  15. Decontamination of Aspergillus flavus and Aspergillus parasiticus spores on hazelnuts via atmospheric pressure fluidized bed plasma reactor.

    PubMed

    Dasan, Beyhan Gunaydin; Mutlu, Mehmet; Boyaci, Ismail Hakki

    2016-01-01

    In this study, an atmospheric pressure fluidized bed plasma (APFBP) system was designed and its decontamination effect on aflatoxigenic fungi (Aspergillus flavus and Aspergillus parasiticus) on the surface of hazelnuts was investigated. Hazelnuts were artificially contaminated with A. flavus and A. parasiticus and then were treated with dry air plasma for up to 5min in the APFBP system at various plasma parameters. Significant reductions of 4.50 log (cfu/g) in A. flavus and 4.19 log (cfu/g) in A. parasiticus were achieved after 5min treatments at 100% V - 25kHz (655W) by using dry air as the plasma forming gas. The decontamination effect of APFBP on A. flavus and A. parasiticus spores inoculated on hazelnuts was increased with the applied reference voltage and the frequency. No change or slight reductions were observed in A. flavus and A. parasiticus load during the storage of plasma treated hazelnuts whereas on the control samples fungi continued to grow under storage conditions (30days at 25°C). Temperature change on hazelnut surfaces in the range between 35 and 90°C was monitored with a thermal camera, and it was demonstrated that the temperature increase taking place during plasma treatment did not have a lethal effect on A. flavus and A. parasiticus spores. The damage caused by APFBP treatment on Aspergillus spp. spores was also observed by scanning electron microscopy. PMID:26398284

  16. Production of chitin deacetylase by Aspergillus flavus in submerged conditions.

    PubMed

    Narayanan, Karthik; Parameswaran, Binod; Pandey, Ashok

    2016-07-01

    Chitosan is a biopolymer obtained by deacetylation of chitin and has been proven to have various applications in industry and biomedicine. Deacetylation of chitin using the enzyme chitin deacetylase (CDA) is favorable in comparison to the hazardous chemical method involving strong alkali and high temperature. A fungal strain producing CDA was isolated from environmental samples collected from coastal regions of South Kerala, India. It was identified as Aspergillus flavus by morphological characteristics and ITS DNA analysis. Nutritional requirement for maximum production of CDA under submerged condition was optimized using statistical methods including Plackett-Burman and response surface methodology central composite design. A 5.98-fold enhancement in CDA production was attained in shake flasks when the fermentation process parameters were used at their optimum levels. The highest CDA activity was 57.69 ± 1.68 U under optimized bioprocess conditions that included 30 g L(-1) glucose, 40 g L(-1) yeast extract, 15 g L(-1) peptone, and 7 g L(-1) MgCl2 at initial media pH of 7 and incubation temperature of 32°C after 48 hr of incubation, while the unoptimized basal medium yielded 9.64 ± 2.04 U. PMID:26474347

  17. Toward elucidation of genetic and functional genetic mechanisms in corn host resistance to Aspergillus flavus infection and aflatoxin contamination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are carcinogenic mycotoxins produced by the fungus Aspergillus flavus. Aflatoxin contamination in pre-harvest corn profusely happens when heat and drought field conditions favor A. flavus colonization. Commercial corn hybrids are generally susceptible to A. flavus infection. An ideal cont...

  18. Stimulation by Hyphopichia burtonii and Bacillus amyloliquefaciens of aflatoxin production by Aspergillus flavus in irradiated maize and rice grains

    SciTech Connect

    Cuero, R.G.; Smith, J.E.; Lacey, J.

    1987-05-01

    Aspergillus flavus was grown on maize and rice extract agars and on irradiated viable cracked maize and rice grains, either in pure culture or in dual culture with wild strains of either Hyphopichia burtonii or Bacillus amyloliquefaciens. Aflatoxin production by A. flavus and its growth and interactions with the other microorganisms were studied at three water activities (a/sub w/) (0.98, 0.95, and 0.90) and two temperatures (25 and 16/sup 0/C). Both H. burtonii and B. amyloliquefaciens markedly stimulated growth and aflotoxin production by A. flavus on cracked maize, especially at 25/sup 0/C and 0.95 and 0.98 a/sub w/. No aflatoxin was detected in pure cultures of A. flavus on cracked rice after 12 days of incubation at 25/sup 0/C, but some was produced by mixed cultures at 16/sup 0/C and 0.98 a/sub w/. The morphological interactions among A. flavus, H. burtonii, and B. amyloliquefaciens were also examined on maize and rice extract agars under similar controlled conditions.

  19. 77 FR 14287 - Aspergillus flavus AF36; Amendment to an Exemption From the Requirement of a Tolerance

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-09

    ...This regulation amends the existing temporary tolerance exemption for Aspergillus flavus AF36 by establishing a permanent exemption from the requirement of a tolerance for residues of the microbial pesticide, Aspergillus flavus AF36, in or on pistachio when applied as an antifungal agent and used in accordance with good agricultural practices. On behalf of the Arizona Cotton Research and......

  20. RNA sequencing of an nsdC mutant reveals global regulation of secondary metabolic gene clusters in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The zinc finger transcription factor nsdC is required for both sexual development and aflatoxin production in the saprophytic fungus Aspergillus flavus. While previous work with an nsdC knockout mutant was conducted in Aspergillus nidulans and A. flavus strain 3357, here we demonstrate perturbations...

  1. Expression of Genes by Aflatoxigenic and Nonaflatoxigenic Strains of Aspergillus flavus Isolated from Brazil Nuts.

    PubMed

    Baquião, Arianne Costa; Rodriges, Aline Guedes; Lopes, Evandro Luiz; Tralamazza, Sabina Moser; Zorzete, Patricia; Correa, Benedito

    2016-08-01

    The aims of the present study were to monitor the production of aflatoxin B1 (AFB1) and mycelial growth, and to evaluate the expression of genes directly and indirectly involved in the biosynthesis of aflatoxins by Aspergillus flavus isolated from Brazil nuts. Six previously identified A. flavus strains were grown on coconut agar at 25°C for up to 10 days. Mycotoxins were separated by high-performance liquid chromatography and fungal growth was measured daily using the diametric mycelial growth rate. Transcriptional analysis was performed by real-time polymerase chain reaction (PCR) after 2 and 7 d of incubation using specific primers (aflR, aflD, aflP, lipase, metalloprotease, and LaeA). Three (50%) of the six A. flavus isolates produced AFB1 (ICB-1, ICB-12, and ICB-54) and three (50%) were not aflatoxigenic (ICB-141, ICB-161, and ICB-198). Aflatoxin production was observed from d 2 of incubation (1.5 ng/g for ICB-54) and increased gradually with time of incubation until d 10 (15,803.6 ng/g for ICB-54). Almost all A. flavus isolates exhibited a similar gene expression pattern after 2 d of incubation (p > 0.10). After 7 d of incubation, the LaeA (p < 0.05) and metalloprotease (p < 0.05) genes were the most expressed by nonaflatoxigenic strains, whereas aflatoxigenic isolates exhibited higher expression of the aflR (p < 0.05) and aflD genes (p < 0.05). Our results suggest that the expression of aflR and aflD is correlated with aflatoxin production in A. flavus and that overexpression of aflR could affect the transcriptional and aflatoxigenic pattern (ICB-54). Elucidation of the molecular mechanisms that regulate the secondary metabolism of toxigenic fungi may permit the rational silencing of the genes involved and consequently the programmed inhibition of aflatoxin production. Knowledge of the conditions, under which aflatoxin genes are expressed, should contribute to the development of innovative and more cost-effective strategies to

  2. A protective endophyte of maize: Acremonium zeae antibiotics inhibitory to Aspergillus flavus and Fusarium verticillioides.

    PubMed

    Wicklow, Donald T; Roth, Shoshannah; Deyrup, Stephen T; Gloer, James B

    2005-05-01

    The maize endophyte Acremonium zeae is antagonistic to kernel rotting and mycotoxin producing fungi Aspergillus flavus and Fusarium verticillioides in cultural tests for antagonism, and interferes with A. flavus infection and aflatoxin contamination of preharvest maize kernels. Chemical studies of an organic extract from maize kernel fermentations of Acremonium zeae (NRRL 13540), which displayed significant antifungal activity against Aspergillus flavus and F. verticillioides, revealed that the metabolites accounting for this activity were two newly reported antibiotics pyrrocidines A and B. Pyrrocidines were detected in fermentation extracts for 12 NRRL cultures of Acremonium zeae isolated from maize kernels harvested in Illinois (4/4 cultures), North Carolina (5/5), Georgia (1/2) and unrecorded locations within the USA (2/2). Pyrrocidine B was detected by LCMSMS in whole symptomatic maize kernels removed at harvest from ears of a commercial hybrid that were wound-inoculated in the milk stage with A. zeae (NRRL 13540) or (NRRL 13541). The pyrrocidines were first reported from the fermentation broth of an unidentified filamentous fungus LL-Cyan426, isolated from a mixed Douglas Fir hardwood forest on Crane Island Preserve, Washington, in 1993. Pyrrocidine A exhibited potent activity against most Gram-positive bacteria, including drug-resistant strains, and was also active against the yeast Candida albicans. In an evaluation of cultural antagonism between 13 isolates of A. zeae in pairings with A. flavus (NRRL 6541) and F. verticillioides (NRRL 25457), A. zeae (NRRL 6415) and (NRRL 34556) produced the strongest reaction, inhibiting both organisms at a distance while continuing to grow through the resulting clear zone at an unchanged rate. Maximum colony diameters for A. zeae (NRRL 6415) and (NRRL 13540), on potato dextrose agar after 14 d, were attained within the range of 25-30 degrees C, with less growth recorded at 15 degrees and 37.5 degrees and no growth at 5

  3. RNA sequencing of an nsdC mutant reveals global regulation of secondary metabolic gene clusters in Aspergillus flavus.

    PubMed

    Gilbert, Matthew K; Mack, Brian M; Wei, Qijian; Bland, John M; Bhatnagar, Deepak; Cary, Jeffrey W

    2016-01-01

    The filamentous fungus, Aspergillus flavus (A. flavus) is an opportunistic pathogen capable of invading a number of crops and contaminating them with toxic secondary metabolites such as aflatoxins. Characterizing the molecular mechanisms governing growth and development of this organism is vital for developing safe and effective strategies for reducing crop contamination. The transcription factor nsdC has been identified as being required for normal asexual development and aflatoxin production in A. flavus. Building on a previous study using a large (L)-sclerotial morphotype A. flavus nsdC mutant we observed alterations in conidiophore development and loss of sclerotial and aflatoxin production using a nsdC mutant of a small (S)-sclerotial morphotype, that normally produces aflatoxin and sclerotia in quantities much higher than the L-morphotype. RNA sequencing analysis of the nsdC knockout mutant and isogenic control strain identified a number of differentially expressed genes related to development and production of secondary metabolites, including aflatoxin, penicillin and aflatrem. Further, RNA-seq data indicating down regulation of aflatrem biosynthetic gene expression in the nsdC mutant correlated with HPLC analyses showing a decrease in aflatrem levels. The current study expands the role of nsdC as a globally acting transcription factor that is a critical regulator of both asexual reproduction and secondary metabolism in A. flavus. PMID:26686623

  4. Control of Aspergillus flavus in maize with plant essential oils and their components.

    PubMed

    Montes-Belmont, R; Carvajal, M

    1998-05-01

    The effects of 11 plant essential oils for maize kernel protection against Aspergillus flavus were studied. Tests were conducted to determine optimal levels of dosages for maize protection, effects of combinations of essential oils, and residual effects and toxicity of essential oils to maize plants. Principal constituents of eight essential oils were tested for ability to protect maize kernels. Essential oils of Cinnamomum zeylanicum (cinnamon), Mentha piperita (peppermint), Ocimum basilicum (basil), Origanum vulgare (origanum), Teloxys ambrosioides (the flavoring herb epazote), Syzygium aromaticum (clove), and Thymus vulgaris (thyme) caused a total inhibition of fungal development on maize kernels. Thymol and o-methoxycinnamaldehyde significantly reduced maize grain contamination. The optimal dosage for protection of maize varied from 3 to 8%. Combinations of C. zeylanicum with the remaining oils gave efficient control. A residual effect of C. zeylanicum was detected after 4 weeks of kernel treatment. No phytotoxic effect on germination and corn growth was detected with any of these oils. PMID:9709236

  5. Global Survey of Canonical Aspergillus flavus G Protein-Coupled Receptors

    PubMed Central

    Affeldt, Katharyn J.; Carrig, Joseph; Amare, Meareg

    2014-01-01

    ABSTRACT G protein-coupled receptors (GPCRs) are transmembrane receptors that relay signals from the external environment inside the cell, allowing an organism to adapt to its surroundings. They are known to detect a vast array of ligands, including sugars, amino acids, pheromone peptides, nitrogen sources, oxylipins, and light. Despite their prevalence in fungal genomes, very little is known about the functions of filamentous fungal GPCRs. Here we present the first full-genome assessment of fungal GPCRs through characterization of null mutants of all 15 GPCRs encoded by the aflatoxin-producing fungus Aspergillus flavus. All strains were assessed for growth, development, ability to produce aflatoxin, and response to carbon sources, nitrogen sources, stress agents, and lipids. Most GPCR mutants were aberrant in one or more response processes, possibly indicative of cross talk in downstream signaling pathways. Interestingly, the biological defects of the mutants did not correspond with assignment to established GPCR classes; this is likely due to the paucity of data for characterized fungal GPCRs. Many of the GPCR transcripts were differentially regulated under various conditions as well. The data presented here provide an extensive overview of the full set of GPCRs encoded by A. flavus and provide a framework for analysis in other fungal species. PMID:25316696

  6. Inhibition of Aspergillus flavus in soil by antagonistic Pseudomonas strains reduces the potential for airborne spore dispersal.

    PubMed

    Palumbo, Jeffrey D; O'Keeffe, Teresa L; Kattan, Ali; Abbas, Hamed K; Johnson, Bobbie J

    2010-06-01

    Pseudomonas chlororaphis strain JP1015 and P. fluorescens strain JP2175 were previously isolated from Mississippi cornfield soil samples and selected for their growth inhibition of Aspergillus flavus in laboratory culture. In this study, the antifungal activity of these bacterial strains against A. flavus in soil coculture was determined. Growth of A. flavus was inhibited up to 100-fold by P. chlororaphis strain JP1015 and up to 58-fold by P. fluorescens strain JP2175 within 3 days following soil coinoculation. A. flavus propagule densities after 16 days remained 7- to 20-fold lower in soil treated with either bacterial strain. Using a bench-scale wind chamber, we demonstrated that treatments of soil with P. chlororaphis strain JP1015 and P. fluorescens strain JP2175 reduced airborne spores dispersed across a 1 m distance by 75- to 1,000-fold and 10- to 50-fold, respectively, depending on soil type and inoculum level. These results suggest that application of these bacterial strains may be effective in reducing soil populations of mycotoxigenic fungi, thereby reducing fungal spore formation, and ultimately reducing the potential for crop plant infection via airborne transmission. PMID:20465408

  7. Isolation and structural elucidation of acidic terpenoid phytoalexins in maize and their interactions with Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plants use a variety of physical and chemical defenses in response to herbivory and pathogen attack. Infection of maize by the fungal pathogen Aspergillus flavus results in the accumulation of aflatoxins, which are among the most detrimental biogenic substances known to man. The majority of maize de...

  8. Intraspecific competition during infection by Aspergillus flavus is influenced by plant host species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Communities of Aspergillus flavus are composed of diverse genotypes that collectively influence incidence and severity of crop aflatoxin contamination. Isolates vary in competitive ability on maize, but empirical data on the extent to which host-specific influences determine outcomes of competition ...

  9. Genes differentially expressed by Aspergillus flavus strains after loss of aflatoxin production by serial transfers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are carcinogenic fungal secondary metabolites produced by Aspergillus flavus and other closely related species. Levels of aflatoxins in agricultural commodities are stringently regulated by many countries and thus aflatoxins are a major concern to both producers and consumers. A cluster...

  10. Crop rotation and soil temperature influence the community structure of Aspergillus flavus in soil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus, the most important cause of aflatoxin contamination has two major morphotypes commonly named S and L strains. Strain S isolates, on average, produce more aflatoxins than the strain L isolates. The S strain has been implicated as the primary causal agent of several contamination e...

  11. Aflatoxigenesis induced in Aspergillus flavus by oxidative stress and reduction by phenolic antioxidants from tree nuts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almonds, pistachios, and walnuts grown in California have an aggregate value of over $3.3 billion, with a large proportion of the crop being exported. However, these tree nuts can be subject to contamination by aflatoxins, metabolites produced primarily by Aspergillus flavus and parasiticus, and im...

  12. RNA-seq analysis of an nsdC mutant in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The C2H2-type transcription factor NsdC (Never in Sexual Development C) has been shown to play a role in asexual development and secondary metabolite production in Aspergillus flavus, an agriculturally relevant, aflatoxin-producing species. The nsdC knoackout mutant demonstrates perturbed morphologi...

  13. Influence of Gene Expression on Variable Aflatoxin Production by Different Strains of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a globally distributed fungus. It causes disease in human and crop plants due to the production of numerous conidia dispersed by air movement and possibly by insects. The fungus is an economically important food contaminant because it produces the most potent natural carcinogen...

  14. 40 CFR 180.1206 - Aspergillus flavus AF36; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... tolerance is established for residues of Aspergillus flavus AF36 in or on corn, field, forage; corn, field, grain; corn, field, stover; corn, field, aspirated grain fractions; corn, sweet, kernel plus cob with husk removed; corn, sweet, forage; corn, sweet, stover; corn, pop, grain; and corn, pop, stover,...

  15. 40 CFR 180.1206 - Aspergillus flavus AF36; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... tolerance is established for residues of Aspergillus flavus AF36 in or on corn, field, forage; corn, field, grain; corn, field, stover; corn, field, aspirated grain fractions; corn, sweet, kernel plus cob with husk removed; corn, sweet, forage; corn, sweet, stover; corn, pop, grain; and corn, pop, stover,...

  16. 40 CFR 180.1206 - Aspergillus flavus AF36; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... tolerance is established for residues of Aspergillus flavus AF36 in or on corn, field, forage; corn, field, grain; corn, field, stover; corn, field, aspirated grain fractions; corn, sweet, kernel plus cob with husk removed; corn, sweet, forage; corn, sweet, stover; corn, pop, grain; and corn, pop, stover,...

  17. Aspergillus flavus Aflatoxin Occurrence and Expression of Aflatoxin Biosynthesis Genes in Soil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The carcinogen, aflatoxin B1 (AFB1) produced by Aspergillus flavus, is a major food safety concern in crops. However, information on AFB1 occurrence in soil and crop residue is scarce. A series of experiments investigated the occurrence of AFB1 in soil and corn residues, and ascertained the ecology ...

  18. Identifying cotton (Gossypium hirsutum L.) genes induced in response to Aspergillus flavus infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination in cottonseed, caused primarily by Aspergillus flavus, is a global concern in terms of food safety and economy. Current strategies to reduce the risk of aflatoxin contamination rely mostly upon biological control with non-aflatoxigenic strains and chemical control measures. ...

  19. CONSTRUCTION OF EXPRESSION CASSETTES TO CONFER RESISTANCE TO ASPERGILLUS FLAVUS IN COTTON

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have been working to develop cotton that is resistant to the fungal pathogen Aspergillus flavus using a genetic engineering approach. Success of this project depends upon the identification of appropriate regulatory elements, as well as structural genes that can be linked to confer a new pathoge...

  20. Potential roles of environmental oxidative stress in aflatoxin production revealed in the Aspergillus flavus transcriptome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination caused by Aspergillus flavus infection in crops is known to be exacerbated primarily by abiotic stresses such as drought stress, and biotic stresses such as arthropod infestation. These stresses result in the production and accumulation of reactive oxygen species (ROS) in the...

  1. Managing and Monitoring of Aspergillus flavus in Corn Using Bioplastic-based Formulation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study, we evaluated the feasibility of bioplastic-based formulations for delivering a non-aflatoxigenic strain of Aspergillus flavus and for monitoring Aspergilli with the final objective of controlling aflatoxin contamination in corn. Field application of inoculated bioplastic granules show...

  2. Hyperspectral image classification and development of fluorescence index for single corn kernels infected with Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic secondary metabolites predominantly produced by the fungi Aspergillus flavus and A. parasiticus. Aflatoxin contaminated corn is toxic to domestic animals when ingested in feed and is a known carcinogen associated with liver and lung cancer in humans. Consequently, aflatoxin leve...

  3. Aspergillus flavus whole genome and EST sequence releases and construction of homologous gene search blast server

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic and carcinogenic secondary metabolites. These compounds, produced by Aspergillus flavus and A. parasiticus, contaminate pre-harvest agricultural crops in the field and post-harvest grains during storage. In order to reduce and eliminate aflatoxin contamination of food and feed...

  4. Resistance to Aspergillus flavus in maize and peanut: Molecular biology, breeding, environmental stress and future perspectives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The colonization of maize (Zea mays L.) and peanut (Arachis hypogaea L.) by the fungal pathogen Aspergillus flavus and A. parasiticus results in the contamination with carcinogenic mycotoxins known as aflatoxins leading to economic losses as well as a potential health threat to human. The interactio...

  5. Identification of novel metabolites from Aspergillus flavus by high resolution and multiple stage mass spectrometry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus contains more than 55 gene clusters which are predicted to encode proteins involved in secondary metabolite production. One of these, cluster 27, contains a polyketide synthase (pks27) gene which encodes a protein that is highly homologous to the aflatoxin cluster PKS. Comparative...

  6. Genome wide association mapping of Aspergillus flavus and aflatoxin accumulation resistance in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Contamination of maize with aflatoxin, produced by the fungus Aspergillus flavus, has severe health and economic consequences. Efforts to reduce aflatoxin accumulation in maize have focused on identifying and selecting germplasm with natural host resistance factors, and several maize lines with sign...

  7. Ear Rot, Aflatoxin Accumulation, and Fungal Biomass in Maize after Inoculation with Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin, a toxin produced by the fungus Aspergillus flavus Link:Fries, occurs naturally in maize (Zea mays L.). Aflatoxin is a potent human carcinogen and is toxic to livestock, pets, and wildlife. When contaminated with aflatoxin, the value of maize grain is markedly reduced. Eight germplasm l...

  8. Isolation of maize soil and rhizosphere bacteria with antagonistic activity against Aspergillus flavus and Fusarium verticillioides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial isolates from Mississippi maize field soil and maize rhizosphere samples were evaluated for their potential as biological control agents against Aspergillus flavus and Fusarium verticillioides. Isolated strains were screened for antagonistic activities in liquid co-culture against A. flav...

  9. Is rachis lignification a deterrent to Aspergillus flavus movement through the developing maize ear?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In previous research, the proteomes of the immature rachis of Aspergillus flavus resistant and susceptible maize inbreds were compared using differential-in-gel electrophoresis (DIGE). One of the proteins that was 66-fold more abundant in the resistant inbred MP313E than in the susceptible inbred S...

  10. The inhibitory effect of Bacillus megaterium on aflatoxin biosynthetic pathway gene expression in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is one of the major fungal mold that colonize peanut in the field and during storage. The impacts to human and animal health and to economy in agriculture and commerce are significant since this mould produces the most potent natural toxins, aflatoxins, which are carcinogenic, mut...

  11. Insights into sexual reproduction in Aspergillus flavus from variation in experimental crosses and natural populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus contaminates many important crops worldwide and is the major producer of aflatoxins, which are cancer-causing secondary metabolites. Biological control is the most effective means of reducing inoculum levels of detrimental aflatoxin-producing fungal pathogens in agricultural syst...

  12. Understanding the Genetics of Regulation of Aflatoxin Production and Aspergillus flavus Development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are polyketide-derived, toxic and carcinogenic secondary metabolites produced primarily by two fungal species, Aspergillus flavus and A. parasiticus, on crops such as corn, peanuts, cottonseed and treenuts. Regulatory guidelines issued by the U.S. Food and Drug Administration (FDA) preve...

  13. Application of biotechnology towards the enhancement of maize resistance to aflatoxin contamination by Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Contamination of maize with aflatoxins by the fungi Aspergillus flavus and A. parasiticus poses serious health hazards to humans and animals worldwide. This important fact and the regulations instituted in many countries to control the occurrence of aflatoxins in foods and feed have stimulated rese...

  14. Transcriptomic analysis reveal diverse responses to environmental oxidative stress in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Drought stress predisposes oilseed crops such as maize and peanut to infection by Aspergillus flavus resulting in their contamination with aflatoxins. Drought stress in plants results in the accumulation of reactive oxygen species (ROS) in their tissues, and these ROS have been shown to stimulate af...

  15. Comparison of Inoculation Methods for Evaluating Maize for Resistance to Aspergillus flavus Infection and Aflatoxin Accumulation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin, the most potent carcinogen found in nature, is produced by the fungus Aspergillus flavus. Aflatoxin occurs naturally in maize, Zea mays L. Growing maize hybrids with genetic resistance to aflatoxin contamination is generally considered a highly desirable way to reduce losses to aflatoxin....

  16. Evaluation of the atoxigenic Aspergillus flavus strain AF36 in pistachio orchards

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The atoxigenic strain Aspergillus flavus AF36, which has been extensively used as a biocontrol agent in commercial corn and cotton fields to reduce aflatoxin contamination, was applied in research pistachio orchards from 2002 to 2005 and in commercial pistachio orchards from 2008 to 2011. AF36 was a...

  17. Aspergillus flavus Genomics as a Tool for Studying the Mechanism of Aflatoxin Formation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a pathogen that infects plants, animals, and humans. It produces the most potent carcinogens, known as aflatoxins, when it infects agricultural crops. In order to devise strategies to control aflatoxin contamination of pre-harvest agricultural crops and post harvest grains du...

  18. Identification of atoxigenic Aspergillus flavus isolates to reduce aflatoxin contamination of maize in Kenya

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acute aflatoxin poisonings (aflatoxicosis) in Kenya have led to the deaths of several hundred people between 2004 and 2006. Etiology of contamination in the outbreak districts (Eastern Province) identified an unusual fungal community structure dominated by the highly toxigenic Aspergillus flavus S s...

  19. Testing the efficacy of eGFP-transformed Aspergillus flavus as biocontrol strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Current biological control methods to prevent pre-harvest aflatoxin contamination of corn, cottonseed, and ground and tree nuts involve field inoculation of non-aflatoxigenic Aspergillus flavus. To date, the efficacy of this approach requires annual reapplication of the biocontrol agent. The reason ...

  20. A maize lectin-like protein with antifungal activity against Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The filamentous fungus, Aspergillus flavus, causes an ear rot on maize and produces a mycotoxin, aflatoxin, in colonized maize kernels. Aflatoxins are carcinogenic to humans and animals upon ingestion. The presence of aflatoxins in food and feed is strictly regulated by several governmental agenci...

  1. Evidence of extensive recombination in the aflatoxin gene cluster of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic compounds produced by several Aspergillus species that contaminate crops worldwide. A. flavus is the most common agent of aflatoxin contamination of corn, peanuts, cottonseed, figs and tree nuts in the US. Extensive studies have elucidated the biochemical and regulatory mechan...

  2. Comparative transcriptome analysis of Aspergillus flavus isolates under different oxidative stresses and culture media

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and aflatoxin contamination in the field are known to be influenced by numerous stress factors, particularly drought and heat stress. However, the purpose of aflatoxin production is unknown. Here, we report transcriptome analyses comprised of 282.6 Gb of sequencing data describing...

  3. New Monomeric Stilbenoids from Peanut (Arachis hypogaea) Seeds Challenged by an Aspergillus flavus Strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two new stilbene derivatives have been isolated from peanut seeds challenged by an Aspergillus flavus strain, along with chiricanine B that has not been reported from peanuts, as well as a stilbenoid that has been known as a synthetic product. The structures of these new putative phytoalexins were d...

  4. Efficacy of water dispersible formulations of biocontrol strains of Aspergillus flavus for aflatoxin management in corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Field experiments were conducted in 2011 and 2012 to evaluate the efficacy of water dispersible granule (WDG) formulations of biocontrol strains of Aspergillus flavus in controlling aflatoxin contamination of corn. In 2011, when aflatoxin was present at very high levels, no WDG treatment provided s...

  5. Characterization of aflatoxigenic Aspergillus flavus and A. parasiticus strain isolates from animal feedstuffs in northeastern Iran

    PubMed Central

    Davari, E; Mohsenzadeh, M; Mohammadi, Gh; Rezaeian-Doloei, R

    2015-01-01

    Aflatoxins are secondary toxic metabolites produced by some Aspergillus spp. particularly, Aspergillus flavus and A. parasiticus that contaminate food and feed. The objective of this study was to evaluate the contamination of feedstuffs with Aspergillus spp. and detect genes involved in the aflatoxin biosynthesis pathway of A. flavus and A. parasiticus isolates. A total of 110 cow feed samples (comprised of silage, concentrate, hay and total mixed ration) from 30 industrial and semi-industrial dairy farms of Khorasan Razavi province, northeastern Iran, were examined using cultural and PCR methods. 68 (61.82%) Aspergillus spp. were isolated from 110 samples of feedstuff. The predominant Aspergillus isolates were A. fumigates (21.81%), followed by A. flavus (17.27%), A. niger (10%), A. parasiticus (8.18%), and A. oryzae (4.54%). Fungal contamination levels of industrial and semi-industrial dairy farm samples were not significantly different (P>0.05). Using four sets of primers, a quadruplex PCR was developed to detect genes (nor1, ver1, omtA and aflR) at different loci coding enzymes in the aflatoxin biosynthesis pathway of A. flavus and A. parasiticus strains. Out of 28 strains of A. flavus and A. parasiticus, 10 isolates (35.71%) showed a quadruplet pattern indicating the important genes involved in the aflatoxin biosynthesis pathway, encoded for functional products. These isolates were confirmed to be aflatoxigenic by Thin Layer Chromatography. 18 isolates (64.29%) had three, two and single molecular patterns. The results obtained by this study show that rapid and specific detection of aflatoxigenic molds is important to ensure the microbiological safety of feedstuffs. PMID:27175167

  6. Toxigenic potentiality of Aspergillus flavus and Aspergillus parasiticus strains isolated from black pepper assessed by an LC-MS/MS based multi-mycotoxin method.

    PubMed

    Yogendrarajah, Pratheeba; Devlieghere, Frank; Njumbe Ediage, Emmanuel; Jacxsens, Liesbeth; De Meulenaer, Bruno; De Saeger, Sarah

    2015-12-01

    A liquid chromatography triple quadrupole tandem mass spectrometry method was developed and validated to determine mycotoxins, produced by fungal isolates grown on malt extract agar (MEA). All twenty metabolites produced by different fungal species were extracted using acetonitrile/1% formic acid. The developed method was applied to assess the toxigenic potentiality of Aspergillus flavus (n = 11) and Aspergillus parasiticus (n = 6) strains isolated from black peppers (Piper nigrum L.) following their growth at 22, 30 and 37 °C. Highest mean radial colony growth rates were observed at 30 °C for A. flavus (5.21 ± 0.68 mm/day) and A. parasiticus (4.97 ± 0.33 mm/day). All of the A. flavus isolates produced aflatoxin B1 and O-methyl sterigmatocystin (OMST) while 91% produced aflatoxin B2 (AFB2) and 82% of them produced sterigmatocystin (STERIG) at 30 °C. Except one, all the A. parasiticus isolates produced all the four aflatoxins, STERIG and OMST at 30 °C. Remarkably high AFB1 was produced by some A. flavus isolates at 22 °C (max 16-40 mg/kg). Production of mycotoxins followed a different trend than that of growth rate of both species. Notable correlations were found between different secondary metabolites of both species; R(2) 0.87 between AFB1 and AFB2 production. Occurrence of OMST could be used as a predictor for AFB1 production. PMID:26338134

  7. Modeling kinetics of aflatoxin production by Aspergillus flavus in maize-based medium and maize grain.

    PubMed

    Garcia, Daiana; Ramos, Antonio J; Sanchis, Vicente; Marín, Sonia

    2013-03-15

    Predictive mycology has dealt mainly with germination, growth and inactivation of fungi while the issue of mycotoxin production remains relatively unexplored. Very few studies provide biomass dry weight/colony size data along with mycotoxin data for the same sample times, thus the ratio mycotoxin accumulation per fungal biomass dry weight/colony size has rarely been reported. For this reason, the objective of the present study was to model the kinetics of mycotoxin production under the assumption of existing both no-growth-associated and growth-associated production. Aspergillus flavus was chosen as a model mycotoxigenic microorganism, and it was grown in maize agar medium and maize grain at 0.90 and 0.99 aw at 25°C. A significant positive correlation (p<0.05) was observed among the biomass responses (colony radius and biomass dry weight) in agar medium and colony radius in maize at both aw levels assayed. The Luedeking-Piret model was used to model AFB1 production and reasonable percentages of variability were explained. Moreover, AFB1 production was in general slightly better predicted through colony area. As conclusion, aflatoxin production may follow a mixed-growth associated trend, confirming that toxin formation does not present a clear delay in relation to growth under certain conditions. PMID:23422844

  8. A survey on distribution and toxigenicity of Aspergillus flavus from indoor and outdoor hospital environments.

    PubMed

    Sepahvand, Asghar; Shams-Ghahfarokhi, Masoomeh; Allameh, Abdolamir; Jahanshiri, Zahra; Jamali, Mojdeh; Razzaghi-Abyaneh, Mehdi

    2011-11-01

    In the present study, genetic diversity and mycotoxin profiles of Aspergillus flavus isolated from air (indoors and outdoors), levels (surfaces), and soils of five hospitals in Southwest Iran were examined. From a total of 146 Aspergillus colonies, 63 isolates were finally identified as A. flavus by a combination of colony morphology, microscopic criteria, and mycotoxin profiles. No Aspergillus parasiticus was isolated from examined samples. Chromatographic analyses of A. flavus isolates cultured on yeast extract-sucrose broth by tip culture method showed that approximately 10% and 45% of the isolates were able to produce aflatoxin B(1) (AFB(1)) and cyclopiazonic acid (CPA), respectively. Around 40% of the isolates produced sclerotia on Czapek-Dox agar. The isolates were classified into four chemotypes based on the ability to produce AF and CPA that majority of them (55.5%) belonged to chemotype IV comprising non-mycotoxigenic isolates. Random amplified polymorphic DNA (RAPD) profiles generated by a combination of four selected primers were used to assess genetic relatedness of 16 selected toxigenic and non-toxigenic isolates. The resulting dendrogram demonstrated the formation of two separate clusters for the A. flavus comprised both mycotoxigenic and non-toxigenic isolates in a random distribution. The obtained results in this study showed that RAPD profiling is a promising and efficient tool to determine intra-specific genetic variation among A. flavus populations from hospital environments. A. flavus isolates, either toxigenic or non-toxigenic, should be considered as potential threats for hospitalized patients due to their obvious role in the etiology of nosocomial aspergillosis. PMID:22083786

  9. Aspergillus flavus diversity on crops and in the environment can be exploited to reduce aflatoxin exposure and improve health

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Humans and animals are exposed to aflatoxins, toxic carcinogenic fungal metabolites, through consumption of contaminated food and feed. Aspergillus flavus, the primary causal agent of crop aflatoxin contamination, is composed of phenotypically and genotypically diverse vegetative compatibility group...

  10. Aspergillus flavus and Fusariumverticillioides Induce Tissue Specific Gene Expression of PRms and UGT in Maize Seed before Fungal Colonization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and Fusariumverticillioides are fungal pathogens that colonize maize seeds and contaminate them with mycotoxins. To investigate the plant microbe interactions, we conducted histological and molecular studies to characterize the internal colonization of maize seed by the two fungal...

  11. Sequence of host contact influences the outcome of competition among Aspergillus flavus isolates during host tissue invasion

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Biological control of aflatoxin contamination by Aspergillus flavus is achieved by competitive exclusion of aflatoxin producers by atoxigenic strains. However, factors dictating the extent to which competitive displacement occurs during host infection are unknown. The role of preemptive exclusion in...

  12. Aspergillus flavus and fusarium verticillioides induce tissue specific gene expression of PRms and UGT in maize kernels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and Fusarium verticillioides are fungal pathogens capable of colonizing maize kernels and contaminating them with mycotoxins. Development of effective control strategies is extremely difficult because pathogenesis and host response remain poorly understood. In this study, we monit...

  13. The master transcription factor MtfA governs aflatoxin production, morphological development, and pathogenicity in the fungus Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus produces a variety of toxic secondary metabolites, among them the aflatoxins (AFs) are the most well-known. These compounds are highly mutagenic and carcinogenic, particularly AFB1. A. flavus is capable of colonizing economically important crops contaminating them with AFs. Molecu...

  14. Evaluation of resistance to aflatoxin contamination in kernels of maize genotypes using a GFP-expressing Aspergillus flavus strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Evaluation of resistance or susceptibility of corn inbreds to infection by Aspergillus flavus was evaluated by a kernel screening assay. A GFP-expressing strain of A. flavus was used to accomplish this study to measure fungal spread and aflatoxin levels in real time. Among the four inbreds tested, ...

  15. Aspergillus flavus Genomics for Development of Strategies to Interrupt Aflatoxin Formation and Discovery of Fungal Enzymes for Biofuel Production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus produces toxic and the most carcinogenic mycotoxins, the aflatoxins. The primary objectives of our A. flavus genomics program are to reduce and eliminate aflatoxin contamination in food and feed and control fungal infection in preharvest crops such as corn, cotton, peanut and tre...

  16. Elucidation of veA Dependent Genes Associated with Aflatoxin and Sclerotial Production in Aspergillus flavus by Functional Genomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aflatoxin-producing fungi, Aspergillus flavus and A. parasiticus, form structures called sclerotia that allow for survival under adverse conditions. Deletion of the veA gene in A. flavus and A. parasiticus blocks production of aflatoxin, as well as sclerotial formation. We used microarray tech...

  17. Toxigenic Aspergillus flavus and other fungi of public health concern in food and organic matter in southwest Nigeria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Six Aspergillus flavus isolates out of 17 fungal isolates were sampled from diverse food and organic matter in southwest Nigeria. All the A. flavus samples produced aflatoxin and cyclopiazonic acid. These six isolates constitute a ready mycobank of toxigenic species for analytical research involving...

  18. Genome sequence of Aspergillus flavus NRRL 3357, a strain that causes aflatoxin contamination of food and feed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination of food and livestock feed results in significant annual crop losses internationally. Aspergillus flavus is the major fungus responsible for this loss. Additionally, A. flavus is the second leading cause of aspergillosis in immune compromised human patients. Here we report th...

  19. Effect of sexual recombination on population diversity in aflatoxin production by Aspergillus flavus and evidence for cryptic heterokaryosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is the major producer of carcinogenic aflatoxins (AFs) in crops worldwide. Natural populations of A. flavus show tremendous variation in AF production, some of which can be attributed to environmental conditions, differential regulation of the AF biosynthetic pathway, and deletio...

  20. Field Assessment of Non-toxigenic Aspergillus flavus Strain K49 in Competitive Displacement of Toxigenic Isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Non-toxigenic strains of Aspergillus flavus offer the potential to control aflatoxin contamination by competitive displacement of indigenous populations of A. flavus colonizing corn grain. Two sets of experiments were conducted to assess the competitiveness of strain K49 when challenged against two...

  1. PR10 expression in maize and its effect on host resistance against Aspergillus flavus infection and aflatoxin production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize (Zea mays L.) is a major crop susceptible to Aspergillus flavus infection and subsequent contamination with aflatoxins, the potent carcinogenic secondary metabolites of the fungus. Protein profiles of maize genotypes resistant and susceptible to A. flavus infection and/or aflatoxin contaminati...

  2. Extracellular Xylanolytic and Pectinolytic Hydrolase Production by Aspergillus flavus Isolates Contributes to Crop Invasion.

    PubMed

    Mellon, Jay E

    2015-08-01

    Several atoxigenic Aspergillus flavus isolates, including some being used as biocontrol agents, and one toxigenic isolate were surveyed for the ability to produce extracellular xylanolytic and pectinolytic hydrolases. All of the tested isolates displayed good production of endoxylanases when grown on a medium utilizing larch xylan as a sole carbon substrate. Four of the tested isolates produced reasonably high levels of esterase activity, while the atoxigenic biocontrol agent NRRL 21882 isolate esterase level was significantly lower than the others. Atoxigenic A. flavus isolates 19, 22, K49, AF36 (the latter two are biocontrol agents) and toxigenic AF13 produced copious levels of pectinolytic activity when grown on a pectin medium. The pectinolytic activity levels of the atoxigenic A. flavus 17 and NRRL 21882 isolates were significantly lower than the other tested isolates. In addition, A. flavus isolates that displayed high levels of pectinolytic activity in the plate assay produced high levels of endopolygalacturonase (pectinase) P2c, as ascertained by isoelectric focusing electrophoresis. Isolate NRRL 21882 displayed low levels of both pectinase P2c and pectin methyl esterase. A. flavus appears capable of producing these hydrolytic enzymes irrespective of aflatoxin production. This ability of atoxigenic isolates to produce xylanolytic and pectinolytic hydrolases mimics that of toxigenic isolates and, therefore, contributes to the ability of atoxigenic isolates to occupy the same niche as A. flavus toxigenic isolates. PMID:26295409

  3. Identification of Aspergillus flavus isolates as potential biocontrol agents of aflatoxin contamination in crops.

    PubMed

    Rosada, L J; Sant'anna, J R; Franco, C C S; Esquissato, G N M; Santos, P A S R; Yajima, J P R S; Ferreira, F D; Machinski, M; Corrêa, B; Castro-Prado, M A A

    2013-06-01

    Aspergillus flavus, a haploid organism found worldwide in a variety of crops, including maize, cottonseed, almond, pistachio, and peanut, causes substantial and recurrent worldwide economic liabilities. This filamentous fungus produces aflatoxins (AFLs) B1 and B2, which are among the most carcinogenic compounds from nature, acutely hepatotoxic and immunosuppressive. Recent efforts to reduce AFL contamination in crops have focused on the use of nonaflatoxigenic A. flavus strains as biological control agents. Such agents are applied to soil to competitively exclude native AFL strains from crops and thereby reduce AFL contamination. Because the possibility of genetic recombination in A. flavus could influence the stability of biocontrol strains with the production of novel AFL phenotypes, this article assesses the diversity of vegetative compatibility reactions in isolates of A. flavus to identify heterokaryon self-incompatible (HSI) strains among nonaflatoxigenic isolates, which would be used as biological controls of AFL contamination in crops. Nitrate nonutilizing (nit) mutants were recovered from 25 A. flavus isolates, and based on vegetative complementation between nit mutants and on the microscopic examination of the number of hyphal fusions, five nonaflatoxigenic (6, 7, 9 to 11) and two nontoxigenic (8 and 12) isolates of A. flavus were phenotypically characterized as HSI. Because the number of hyphal fusions is reduced in HSI strains, impairing both heterokaryon formation and the genetic exchanges with aflatoxigenic strains, the HSI isolates characterized here, especially isolates 8 and 12, are potential agents for reducing AFL contamination in crops. PMID:23726204

  4. Extracellular Xylanolytic and Pectinolytic Hydrolase Production by Aspergillus flavus Isolates Contributes to Crop Invasion

    PubMed Central

    Mellon, Jay E.

    2015-01-01

    Several atoxigenic Aspergillus flavus isolates, including some being used as biocontrol agents, and one toxigenic isolate were surveyed for the ability to produce extracellular xylanolytic and pectinolytic hydrolases. All of the tested isolates displayed good production of endoxylanases when grown on a medium utilizing larch xylan as a sole carbon substrate. Four of the tested isolates produced reasonably high levels of esterase activity, while the atoxigenic biocontrol agent NRRL 21882 isolate esterase level was significantly lower than the others. Atoxigenic A. flavus isolates 19, 22, K49, AF36 (the latter two are biocontrol agents) and toxigenic AF13 produced copious levels of pectinolytic activity when grown on a pectin medium. The pectinolytic activity levels of the atoxigenic A. flavus 17 and NRRL 21882 isolates were significantly lower than the other tested isolates. In addition, A. flavus isolates that displayed high levels of pectinolytic activity in the plate assay produced high levels of endopolygalacturonase (pectinase) P2c, as ascertained by isoelectric focusing electrophoresis. Isolate NRRL 21882 displayed low levels of both pectinase P2c and pectin methyl esterase. A. flavus appears capable of producing these hydrolytic enzymes irrespective of aflatoxin production. This ability of atoxigenic isolates to produce xylanolytic and pectinolytic hydrolases mimics that of toxigenic isolates and, therefore, contributes to the ability of atoxigenic isolates to occupy the same niche as A. flavus toxigenic isolates. PMID:26295409

  5. Detection of Aspergillus flavus in stored peanuts using real-time PCR and the expression of aflatoxin genes in toxigenic and atoxigenic A. flavus isolates.

    PubMed

    Mahmoud, Mohamed A

    2015-04-01

    Aspergillus flavus is the main species from section Flavi responsible for aflatoxin accumulation in stored peanuts. Rapid methods to detect A. flavus could help to prevent aflatoxins from entering the food chain. A real-time polymerase chain reaction (RTi-PCR) assay was standardized for rapid, specific, and sensitive detection of A. flavus in stored peanuts. A. flavus was detected in 53.6% and 50% of peanut samples by RTi-PCR and A. flavus and Aspergillus parasiticus agar culture, respectively, with 95% agreement between them. Twenty-two A. flavus isolates were screened using high-performance liquid chromatography for their capacity to produce aflatoxin AFB1 (B1). B1 was produced by >72% of the isolates. Sixteen isolates produced B1 at concentrations ranging from 1.64 to 109.18 μg/mL. Four aflatoxin biosynthetic pathway genes (aflD, aflM, aflP, and aflQ) were evaluated using PCR and reverse-transcription PCR in 22 A. flavus isolates from peanut kernels with the aim of rapidly and accurately differentiating toxigenic and atoxigenic isolates. The PCR amplification of genes did not correlate with aflatoxin production capability. The expression of aflD and aflQ was a good marker for differentiating toxigenic from atoxigenic isolates. PMID:25621617

  6. Effect of climate change on Aspergillus flavus and aflatoxin B1 production

    PubMed Central

    Medina, Angel; Rodriguez, Alicia; Magan, Naresh

    2014-01-01

    This review considers the available information on the potential impact of key environmental factors and their interactions on the molecular ecology, growth and aflatoxin production by Aspergillus flavus in vitro and in maize grain. The recent studies which have been carried out to examine the impact of water activity × temperature on aflatoxin biosynthesis and phenotypic aflatoxin production are examined. These have shown that there is a direct relationship between the relative expression of key regulatory and structural genes under different environmental conditions which correlate directly with aflatoxin B1 production. A model has been developed to integrate the relative expression of 10 biosynthetic genes in the pathway, growth and aflatoxin B1 (AFB1) production which was validated under elevated temperature and water stress conditions. The effect of interacting conditions of aw × temperature × elevated CO2 (2 × and 3 × existing levels) are detailed for the first time. This suggests that while such interacting environmental conditions have little effect on growth they do have a significant impact on aflatoxin biosynthetic gene expression (structural aflD and regulatory aflR genes) and can significantly stimulate the production of AFB1. While the individual factors alone have an impact, it is the combined effect of these three abiotic factors which have an impact on mycotoxin production. This approach provides data which is necessary to help predict the real impacts of climate change on mycotoxigenic fungi. PMID:25101060

  7. Antifungal activity of extracts of Rosmarinus officinalis and Thymus vulgaris against Aspergillus flavus and A. ochraceus.

    PubMed

    Centeno, S; Calvo, M A; Adelantado, C; Figueroa, S

    2010-05-01

    The antifungal activity of ethanolic extracts of Rosmarinus officinalis and Thymus vulgaris were tested against strains of Aspergillus flavus and A. ochraceus, since these two species are common contaminants of cereals and grains and are able to produce and accumulate mycotoxins. The methodology used is based on measuring the inhibition halos produced by discs impregnated with the extracts and establishing their Minimum Inhibitory Concentration (MIC) as well as the Minimum Fungicide Concentration (MFC). The results obtained suggest that the assayed extracts affect the proper development of A. flavus and A. ochraceus; leading to a lower MIC (1200 ppm) and MFC (2400 ppm) for T. vulgaris extract against A. ochraceus than against A. flavus. The results show, that the extracts of Rosmarinus officinalis and Thymus vulgaris used at low concentrations could have significant potential for the biological control of fungi in foodstuffs. PMID:20973400

  8. Molecular characterization of aflatoxigenic and non-aflatoxigenic Aspergillus flavus isolates collected from corn grains.

    PubMed

    Mahmoud, M A; Ali, H M; El-Aziz, A R M; Al-Othman, M R; Al-Wadai, A S

    2014-01-01

    Twelve species from six fungal genera were found to be associated with corn (Zea mays L.) grain samples collected from three main regions of Saudi Arabia. The average frequencies of the most common genera were Aspergillus (11.4%), Fusarium (9.5%), Penicillium (5.1%), and Alternaria (5.8%). Fifteen isolates of Aspergillus flavus were screened by HPLC for their ability to produce aflatoxins (AF). The percentage of aflatoxigenic A. flavus isolates was 53%. Eight isolates produced AF, at concentrations ranging 0.7-2.9 ppb. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) molecular markers were used to genetically characterize isolates of A. flavus and to discriminate between the aflatoxigenic and non-aflatoxigenic isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus population, which was useful for genetic characterization. The clustering in the RAPD and ISSR dendrograms obtained was unrelated to geographic origin. The RAPD and ISSR markers could not discriminate between aflatoxigenic and non-aflatoxigenic isolates, but the ISSR primers were somewhat better. PMID:25501147

  9. Nutritional changes in powdered red pepper upon in vitro infection of Aspergillus flavus

    PubMed Central

    Tripathi, Smita; Mishra, H.N.

    2009-01-01

    Quantitative losses in various biochemical constituents like capsaicin, carotenes, ascorbic acid, polyphenols, mineral matter, sugars (soluble and insoluble), protein and fat were estimated after the successful growth of Aspergillus flavus for 30 days on powdered red pepper. The fungal biomass was measured by ergosterol content and Aflatoxin B1 by HPLC. Amongst the various nutritional constituents evaluated for nutritional losses and changes the highest nutritional loss was reported in total carotenoids (88.55%) followed by total sugars (85.5%). The protein content of the infected sample increased from 18.01% to 23%. The nutritional profile of chilli powder (Capsicum annum var. sannam L.) shows highest share of total soluble sugars (32.89%) and fiber content (21.05%), followed by protein (18.01%) and fat (13.32%) making it an ideal solid- substrate for mould growth. At the end of incubation the fungal biomass was 192. 25 mg / 100 gram powder, total plate count 17.5 X 10 4 CFU/g and Aflatoxin B1 content was 30.06 μg / kg. PMID:24031333

  10. The Mechanism of Antifungal Action of Essential Oil from Dill (Anethum graveolens L.) on Aspergillus flavus

    PubMed Central

    Tian, Jun; Ban, Xiaoquan; Zeng, Hong; He, Jingsheng; Chen, Yuxin; Wang, Youwei

    2012-01-01

    The essential oil extracted from the seeds of dill (Anethum graveolens L.) was demonstrated in this study as a potential source of an eco-friendly antifungal agent. To elucidate the mechanism of the antifungal action further, the effect of the essential oil on the plasma membrane and mitochondria of Aspergillus flavus was investigated. The lesion in the plasma membrane was detected through flow cytometry and further verified through the inhibition of ergosterol synthesis. The essential oil caused morphological changes in the cells of A. flavus and a reduction in the ergosterol quantity. Moreover, mitochondrial membrane potential (MMP), acidification of external medium, and mitochondrial ATPase and dehydrogenase activities were detected. The reactive oxygen species (ROS) accumulation was also examined through fluorometric assay. Exposure to dill oil resulted in an elevation of MMP, and in the suppression of the glucose-induced decrease in external pH at 4 µl/ml. Decreased ATPase and dehydrogenase activities in A. flavus cells were also observed in a dose-dependent manner. The above dysfunctions of the mitochondria caused ROS accumulation in A. flavus. A reduction in cell viability was prevented through the addition of L-cysteine, which indicates that ROS is an important mediator of the antifungal action of dill oil. In summary, the antifungal activity of dill oil results from its ability to disrupt the permeability barrier of the plasma membrane and from the mitochondrial dysfunction-induced ROS accumulation in A. flavus. PMID:22272289

  11. Degeneration of aflatoxin gene clusters in Aspergillus flavus from Africa and North America.

    PubMed

    Adhikari, Bishwo N; Bandyopadhyay, Ranajit; Cotty, Peter J

    2016-12-01

    Aspergillus flavus is the most common causal agent of aflatoxin contamination of food and feed. However, aflatoxin-producing potential varies widely among A. flavus genotypes with many producing no aflatoxins. Some non-aflatoxigenic genotypes are used as biocontrol agents to prevent contamination. Aflatoxin biosynthesis genes are tightly clustered in a highly conserved order. Gene deletions and presence of single nucleotide polymorphisms (SNPs) in aflatoxin biosynthesis genes are often associated with A. flavus inability to produce aflatoxins. In order to identify mechanisms of non-aflatoxigenicity in non-aflatoxigenic genotypes of value in aflatoxin biocontrol, complete cluster sequences of 35 A. flavus genotypes from Africa and North America were analyzed. Inability of some genotypes to produce aflatoxin resulted from deletion of biosynthesis genes. In other genotypes, non-aflatoxigenicity originated from SNP formation. The process of degeneration differed across the gene cluster; genes involved in early biosynthesis stages were more likely to be deleted while genes involved in later stages displayed high frequencies of SNPs. Comparative analyses of aflatoxin gene clusters provides insight into the diversity of mechanisms of non-aflatoxigenicity in A. flavus genotypes used as biological control agents. The sequences provide resources for both diagnosis of non-aflatoxigenicity and monitoring of biocontrol genotypes during biopesticide manufacture and in the environment. PMID:27576895

  12. Loss of msnA, a putative stress regulatory gene, in Aspergillus parasiticus and Aspergillus flavus increased production of conidia, aflatoxins and kojic acid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Production of the harmful carcinogenic aflatoxins by Aspergillus parasiticus and Aspergillus flavus has been postulated to be a mechanism to relieve oxidative stress. The msnA gene, the ortholog of Saccharomyces cerevisiae MSN2 associated with multi-stress response, of the two species was disrupted....

  13. Genome Sequences of Eight Aspergillus flavus spp. and One A. parasiticus sp., Isolated from Peanut Seeds in Georgia

    PubMed Central

    Wang, Xinye Monica; Palencia, Edwin R.

    2016-01-01

    Aspergillus flavus and A. parasiticus fungi produce carcinogenic mycotoxins in peanut seeds, causing considerable impact on both human health and the economy. Here, we report nine genome sequences of Aspergillus spp., isolated from Georgia peanut seeds in 2014. The information obtained will lead to further biodiversity studies that are essential for developing control strategies. PMID:27081142

  14. Aspergillus flavus biomass in maize estimated by quantitative real-time polymerase chain reaction is strongly correlated with aflatoxin concentration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus causes Aspergillus ear rot of maize and produces aflatoxins. There are published assertions that resistance to aflatoxin accumulation and pathogen colonization are distinct traits in maize. However, the levels of colonization are difficult to characterize for a pathogen such as ...

  15. Genome Sequences of Eight Aspergillus flavus spp. and One A. parasiticus sp., Isolated From Peanut Seeds in Georgia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and A. parasiticus fungi, carcinogen-mycotoxins producers, infect peanut seeds, causing considerable impact on both human health and the economy. Here we report 9 genome sequences of Aspergillus spp. isolated from peanut seeds. The information obtained will allow conducting biodiv...

  16. Clustered Genes Involved in Cyclopiazonic Acid Production are Next to the Aflatoxin Biosynthesis Gene Cluster in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cyclopiazonic acid (CPA), an indole-tetramic acid toxin, is produced by many species of Aspergillus and Penicillium. In addition to CPA Aspergillus flavus produces polyketide-derived carcinogenic aflatoxins (AFs). AF biosynthesis genes form a gene cluster in a subtelomeric region. Isolates of A. fla...

  17. A Caleosin-Like Protein with Peroxygenase Activity Mediates Aspergillus flavus Development, Aflatoxin Accumulation, and Seed Infection.

    PubMed

    Hanano, Abdulsamie; Almousally, Ibrahem; Shaban, Mouhnad; Blee, Elizabeth

    2015-09-01

    Caleosins are a small family of calcium-binding proteins endowed with peroxygenase activity in plants. Caleosin-like genes are present in fungi; however, their functions have not been reported yet. In this work, we identify a plant caleosin-like protein in Aspergillus flavus that is highly expressed during the early stages of spore germination. A recombinant purified 32-kDa caleosin-like protein supported peroxygenase activities, including co-oxidation reactions and reduction of polyunsaturated fatty acid hydroperoxides. Deletion of the caleosin gene prevented fungal development. Alternatively, silencing of the gene led to the increased accumulation of endogenous polyunsaturated fatty acid hydroperoxides and antioxidant activities but to a reduction of fungal growth and conidium formation. Two key genes of the aflatoxin biosynthesis pathway, aflR and aflD, were downregulated in the strains in which A. flavus PXG (AfPXG) was silenced, leading to reduced aflatoxin B1 production in vitro. Application of caleosin/peroxygenase-derived oxylipins restored the wild-type phenotype in the strains in which AfPXG was silenced. PXG-deficient A. flavus strains were severely compromised in their capacity to infect maize seeds and to produce aflatoxin. Our results uncover a new branch of the fungal oxylipin pathway and may lead to the development of novel targets for controlling fungal disease. PMID:26116672

  18. A Caleosin-Like Protein with Peroxygenase Activity Mediates Aspergillus flavus Development, Aflatoxin Accumulation, and Seed Infection

    PubMed Central

    Almousally, Ibrahem; Shaban, Mouhnad; Blee, Elizabeth

    2015-01-01

    Caleosins are a small family of calcium-binding proteins endowed with peroxygenase activity in plants. Caleosin-like genes are present in fungi; however, their functions have not been reported yet. In this work, we identify a plant caleosin-like protein in Aspergillus flavus that is highly expressed during the early stages of spore germination. A recombinant purified 32-kDa caleosin-like protein supported peroxygenase activities, including co-oxidation reactions and reduction of polyunsaturated fatty acid hydroperoxides. Deletion of the caleosin gene prevented fungal development. Alternatively, silencing of the gene led to the increased accumulation of endogenous polyunsaturated fatty acid hydroperoxides and antioxidant activities but to a reduction of fungal growth and conidium formation. Two key genes of the aflatoxin biosynthesis pathway, aflR and aflD, were downregulated in the strains in which A. flavus PXG (AfPXG) was silenced, leading to reduced aflatoxin B1 production in vitro. Application of caleosin/peroxygenase-derived oxylipins restored the wild-type phenotype in the strains in which AfPXG was silenced. PXG-deficient A. flavus strains were severely compromised in their capacity to infect maize seeds and to produce aflatoxin. Our results uncover a new branch of the fungal oxylipin pathway and may lead to the development of novel targets for controlling fungal disease. PMID:26116672

  19. Aspergillus flavus-Induced Brain Abscess in an Immunocompetent Child: Case report.

    PubMed

    Al-Maskari, Nawal; Hussain, Ibrahim; Jumaa, Suleiman; Al-Shail, Essam A

    2016-05-01

    Intracranial aspergillosis is an extremely rare manifestation of invasive aspergillosis in immunocompetent children and is associated with high morbidity and mortality. We report a 12-year-old immunocompetent male child who was referred to the King Faisal Specialist Hospital & Research Centre, Riyadh, Saudi Arabia, in May 2010 after a sudden-onset headache and loss of consciousness. Brain imaging revealed a large right space-occupying occipital lesion and the patient underwent a craniotomy and resection. Histopathology of the lesion revealed necrotising granulomatous fungal encephalitis with many hyphae engulfed by multinucleated giant histiocytes. Two days later, a computed tomography scan showed debulking of the fungal mass and the patient was discharged on oral voriconazole. However, imaging at a six-week follow-up showed progression of the abnormality. A residual or persistent fungal brain lesion was suspected. Further neurosurgical resection of the lesion was performed and cultures showed growth of Aspergillus flavus. The patient was treated successfully with antifungal therapy over the following two years. PMID:27226920

  20. Cryptic Sexuality Influences Aflatoxigenicity in Aspergillus parasiticus and A. flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascomycetous fungi of the genus Aspergillus comprise a wide variety of species of biotechnological importance as well as pathogens and toxin producers. Recent studies report A. fumigatus to be heterothallic and possibly undergoing sexual reproduction. We therefore investigated whether compatible mat...

  1. Molecular characterisation of Aspergillus flavus isolates from peanut fields in India using AFLP

    PubMed Central

    Singh, Diwakar; Radhakrishnan, T.; Kumar, Vinod; Bagwan, N.B.; Basu, M.S.; Dobaria, J.R.; Mishra, Gyan P.; Chanda, S.V.

    2015-01-01

    Aflatoxin contamination of peanut, due to infection by Aspergillus flavus, is a major problem of rain-fed agriculture in India. In the present study, molecular characterisation of 187 Aspergillus flavus isolates, which were sampled from the peanut fields of Gujarat state in India, was performed using AFLP markers. On a pooled cluster analysis, the markers could successfully discriminate among the ‘A’, ‘B’ and ‘G’ group A. flavus isolates. PCoA analysis also showed equivalent results to the cluster analysis. Most of the isolates from one district could be clustered together, which indicated genetic similarity among the isolates. Further, a lot of genetic variability was observed within a district and within a group. The results of AMOVA test revealed that the variance within a population (84%) was more than that between two populations (16%). The isolates, when tested by indirect competitive ELISA, showed about 68.5% of them to be atoxigenic. Composite analysis between the aflatoxin production and AFLP data was found to be ineffective in separating the isolate types by aflatoxigenicity. Certain unique fragments, with respect to individual isolates, were also identified that may be used for development of SCAR marker to aid in rapid and precise identification of isolates. PMID:26413047

  2. Molecular characterization of Aspergillus flavus and aflatoxin contamination of wheat grains from Saudi Arabia.

    PubMed

    Al-Wadai, A S; Al-Othman, M R; Mahmoud, M A; Abd El-Aziz, A R M

    2013-01-01

    Twelve species belonging to six fungal genera were found to be associated with wheat (Triticum aestivum L.) grain samples collected from three main regions in Saudi Arabia. The most common genera (average frequency) were Aspergillus (14.3%), Fusarium (29.1%), Penicillium (9.3%), and Alternaria (8.2%). Nineteen isolates of Aspergillus flavus were screened for their ability to produce aflatoxins using HPLC. Thirteen isolates produced aflatoxins ranging from 0.5 to 2.6 µg/kg. Inter-simple sequence repeats (ISSR), and random amplified polymorphic DNA (RAPD) molecular markers were used, with the aim of genetically characterizing strains of A. flavus to discriminate between aflatoxigenic and non-aflatoxigenic isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus population, useful for genetic characterization. Clustering based on RAPD and ISSR dendograms was unrelated to geographic origin. RAPD and ISSR markers were not suitable to discriminate aflatoxigenic and non-aflatoxigenic isolates, but ISSR primers were better compared to RAPD. PMID:24065675

  3. Seventeen years of subcutaneous infection by Aspergillus flavus; eumycetoma confirmed by immunohistochemistry.

    PubMed

    Ahmed, Sarah A; Abbas, Manal A; Jouvion, Gregory; Al-Hatmi, Abdullah M S; de Hoog, G Sybren; Kolecka, Anna; Mahgoub, El Sheikh

    2015-12-01

    Chronic subcutaneous infections caused by Aspergillus species are considered to be extremely rare. Because these fungi are among the most common laboratory contaminants, their role as eumycetoma causative agents is difficult to ascertain. Here, we report the first case of A. flavus eumycetoma confirmed by isolation, molecular identification and immunohistochemical analysis. Patient was a 55-year-old male from Sudan suffering from eumycetoma on his left foot for a period of 17 years. He developed swelling, sinuses and white grain discharge was observed. He has been operated nine times and was treated with several regimens of ketoconazole and itraconazole without improvement. Initial diagnosis based on histology and radiology was Scedosporium eumycetoma. However, examination of the biopsy revealed A. flavus, which was identified by molecular analysis and MALDI-TOF MS. Immunohistochemistry using antibody directed against Aspergillus species was positive. Because of the earlier treatment failures with ketoconazole and itraconazole, therapy with voriconazole was initiated. However, in vitro susceptibility testing yielded a lower Minimum Inhibitory Concentration (MIC) value for itraconazole (0.25 μg ml(-1) ) than for voriconazole (1 μg ml(-1) ). Based on the presented results, A. flavus can be considered as one of the agents of white-grain eumycetoma. PMID:26497138

  4. The DmtA methyltransferase contributes to Aspergillus flavus conidiation, sclerotial production, aflatoxin biosynthesis and virulence.

    PubMed

    Yang, Kunlong; Liang, Linlin; Ran, Fanlei; Liu, Yinghang; Li, Zhenguo; Lan, Huahui; Gao, Peili; Zhuang, Zhenhong; Zhang, Feng; Nie, Xinyi; Kalayu Yirga, Shimuye; Wang, Shihua

    2016-01-01

    DNA methylation is essential for epigenetic regulation of gene transcription and development in many animals, plants and fungi. We investigated whether DNA methylation plays a role in the development and secondary metabolism of Aspergillus flavus, identified the DmtA methyltransferase from A. flavus, and produced a dmtA knock-out mutant by replacing the dmtA coding sequence with the pyrG selectable marker. The A. flavus dmtA null mutant lines produced white fluffy mycelium in liquid medium, and displayed a slightly flavescent conidial pigmentation compared with the normal yellow of the wild-type strain when grown on agar. The ΔdmtA lines exhibited decreased conidiation and aflatoxin (AF) biosynthesis, compared with the wild-type line, suggesting that the DmtA knock-out affected the transcriptional level of genes in the AF cluster. In particular, sclerotia development and host colonization were altered in the dmtA null mutants. Green fluorescent protein tagging at the C-terminus of DmtA showed that DmtA localized to the nucleus and cytoplasm. DNA methylation content measurements in the dmtA mutants revealed no widespread DNA methylation in the mutants or wild-type lines. Thus, our findings suggest that DmtA, apart from being a C-5 cytosine methyltransferase in A. flavus, contributes to asexual development, aflatoxin biosynthesis, sclerotial production and virulence. PMID:26979781

  5. Inactivation of Aspergillus flavus in drinking water after treatment with UV irradiation followed by chlorination.

    PubMed

    Al-Gabr, Hamid Mohammad; Zheng, Tianling; Yu, Xin

    2013-10-01

    The disinfection process for inactivating microorganisms at drinking water treatment plants is aimed for safety of drinking water for humans from a microorganism, such as bacteria, viruses, algae, fungi by using chlorination, ozonation, UV irradiation, etc. In the present study, a combination of two disinfectants, UV irradiation followed by chlorination, was evaluated for inactivating Aspergillus flavus under low contact time and low dosage of UV irradiation. The results indicated an inverse correlation between the inactivation of A. flavus by using UV irradiation only or chlorination alone. By using UV radiation, the 2 log10 control of A. flavus was achieved after 30 s of irradiation, while chlorination was observed to be more effective than UV, where the 2 log was achieved at chlorine concentration of 0.5, 1, 2 and 3 mg/l, in contact time of 60, 5, 1 and 1 min, respectively. However, combined use (UV irradiation followed by chlorination) was more effective than using either UV or chlorination alone; 5 s UV irradiation followed by chlorination produced 4 log10 reduction of A. flavus at chlorine concentrations of 2 and 3 mg/l under a contact time of 15 min. The results indicated that efficiency of UV irradiation improves when followed by chlorination at low concentrations. PMID:23831798

  6. Aflatoxins in Rice Artificially Contaminated with Aflatoxin-producing Aspergillus flavus under Natural Storage in Japan.

    PubMed

    Sugihara, Satoshi; Doi, Hiroyuki; Kato, Masahiko; Mitoh, Yoshihiro; Tsuda, Toshihide; Ikeda, Satoru

    2016-06-01

    Aflatoxin (AFT) contamination is frequent in foods grown in tropical regions, including rice. Although AFTs are generally not found in temperate-region foods, global warming has affected typical temperate-region climates, potentially permitting the contamination of foods with AFT-producing Aspergillus flavus (A. flavus). Here we investigated the AFT production in rice during storage under natural climate conditions in Japan. We examined AFTs in brown rice and rough rice artificially contaminated with A. flavus for 1 year in Japan, and we subjected AFTs in white rice to the same treatment in airtight containers and examined the samples in warm and cold seasons, simulating the storage of white rice in general households. In the brown rice, AFTs increased after 2 months (March) and peaked after 9 months (October). The AFT contamination in the rough rice was minimal. After the polishing and cooking of the brown rice, AFTs were undetectable. In the white rice stored in airtight containers, AFTs increased after 1 month (August) and peaked after 2 months (September). Minimal AFTs were detected in the cold season. Thus, AFT contamination in rice may occur in temperate regions following A. flavus contamination. The storage of rice as rough rice could provide be useful for avoiding AFT contamination. PMID:27339205

  7. Role of Oxidative Stress in Sclerotial Differentiation and Aflatoxin B1 Biosynthesis in Aspergillus flavus

    PubMed Central

    Grintzalis, Konstantinos; Vernardis, Spyros I.; Klapa, Maria I.

    2014-01-01

    We show here that oxidative stress is involved in both sclerotial differentiation (SD) and aflatoxin B1 biosynthesis in Aspergillus flavus. Specifically, we observed that (i) oxidative stress regulates SD, as implied by its inhibition by antioxidant modulators of reactive oxygen species and thiol redox state, and that (ii) aflatoxin B1 biosynthesis and SD are comodulated by oxidative stress. However, aflatoxin B1 biosynthesis is inhibited by lower stress levels compared to SD, as shown by comparison to undifferentiated A. flavus. These same oxidative stress levels also characterize a mutant A. flavus strain, lacking the global regulatory gene veA. This mutant is unable to produce sclerotia and aflatoxin B1. (iii) Further, we show that hydrogen peroxide is the main modulator of A. flavus SD, as shown by its inhibition by both an irreversible inhibitor of catalase activity and a mimetic of superoxide dismutase activity. On the other hand, aflatoxin B1 biosynthesis is controlled by a wider array of oxidative stress factors, such as lipid hydroperoxide, superoxide, and hydroxyl and thiyl radicals. PMID:25002424

  8. The DmtA methyltransferase contributes to Aspergillus flavus conidiation, sclerotial production, aflatoxin biosynthesis and virulence

    PubMed Central

    Yang, Kunlong; Liang, Linlin; Ran, Fanlei; Liu, Yinghang; Li, Zhenguo; Lan, Huahui; Gao, Peili; Zhuang, Zhenhong; Zhang, Feng; Nie, Xinyi; Kalayu Yirga, Shimuye; Wang, Shihua

    2016-01-01

    DNA methylation is essential for epigenetic regulation of gene transcription and development in many animals, plants and fungi. We investigated whether DNA methylation plays a role in the development and secondary metabolism of Aspergillus flavus, identified the DmtA methyltransferase from A. flavus, and produced a dmtA knock-out mutant by replacing the dmtA coding sequence with the pyrG selectable marker. The A. flavus dmtA null mutant lines produced white fluffy mycelium in liquid medium, and displayed a slightly flavescent conidial pigmentation compared with the normal yellow of the wild-type strain when grown on agar. The ΔdmtA lines exhibited decreased conidiation and aflatoxin (AF) biosynthesis, compared with the wild-type line, suggesting that the DmtA knock-out affected the transcriptional level of genes in the AF cluster. In particular, sclerotia development and host colonization were altered in the dmtA null mutants. Green fluorescent protein tagging at the C-terminus of DmtA showed that DmtA localized to the nucleus and cytoplasm. DNA methylation content measurements in the dmtA mutants revealed no widespread DNA methylation in the mutants or wild-type lines. Thus, our findings suggest that DmtA, apart from being a C-5 cytosine methyltransferase in A. flavus, contributes to asexual development, aflatoxin biosynthesis, sclerotial production and virulence. PMID:26979781

  9. 75 FR 9596 - Notice of Filing of a Pesticide Petition for Residues of a Aspergillus flavus AF36 on Corn Food...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-03

    ... From the Federal Register Online via the Government Publishing Office ENVIRONMENTAL PROTECTION AGENCY Notice of Filing of a Pesticide Petition for Residues of a Aspergillus flavus AF36 on Corn Food... residues of the antifungal ] agent, Aspergillus flavus AF36, in or on corn food and feed commodities....

  10. Antimicrobial effects of ionizing radiation on artificially and naturally contaminated cacao beans. [Aspergillus flavus; Penicillium citrinum

    SciTech Connect

    Restaino, L.; Myron, J.J.J.; Lenovich, L.M.; Bills, S.; Tscherneff, K.

    1984-04-01

    With an initial microbial level of ca. 10/sup 7/ microorganisms per g of Ivory Coast cacao beans, 5 kGy of gamma radiation from a Co/sup 60/ source under an atmosphere of air reduced the microflora per g by 2.49 and 3.03 logs at temperatures of 35 and 50/sup 0/C, respectively. Bahia cacao beans were artificially contaminated with dried spores of Aspergillus flavus and Penicillium citrinum, giving initial fungal levels of 1.9 x 10/sup 4/ and 1.4 x 10/sup 3/ spores per g of whole Bahia cacao beans, respectively. The average D/sub 10/ values for A. flavus and P. citrinum spores on Bahia cacao beans were 0.66 and 0.88 kGy, respectively. 12 references.

  11. Degradation of polyurethane by Aspergillus flavus (ITCC 6051) isolated from soil.

    PubMed

    Mathur, Garima; Prasad, Ramasare

    2012-07-01

    The present study deals with the isolation of fungi from soil with the ability to degrade polyurethane (PU). A pure fungal isolate was analyzed for its ability to utilize PU as a sole carbon source in shaking culture for 30 days. Incubation of PU with Aspergillus flavus resulted in 60.6% reduction in weight of PU. The scanning electron microscopy and Fourier transform infrared spectroscopy (FTIR) results showed certain changes on the surface of PU film and formation of some new intermediate products after polymer breakdown. Thermogravimetric curves showed changes between the thermal behavior of the samples that were inoculated with A. flavus and control. FTIR spectra showed detectable changes in control and incubated samples, suggesting that degradation occurs, with the decreased intensity of band at 1,715 cm(-1), corresponding to ester linkages. We have identified an extracellular esterase activity which might be responsible for the polyurethanolytic activity. PMID:22367637

  12. Aspergillus flavus infection induces transcriptional and physical changes in developing maize kernels

    PubMed Central

    Dolezal, Andrea L.; Shu, Xiaomei; OBrian, Gregory R.; Nielsen, Dahlia M.; Woloshuk, Charles P.; Boston, Rebecca S.; Payne, Gary A.

    2014-01-01

    Maize kernels are susceptible to infection by the opportunistic pathogen Aspergillus flavus. Infection results in reduction of grain quality and contamination of kernels with the highly carcinogenic mycotoxin, aflatoxin. To understanding host response to infection by the fungus, transcription of approximately 9000 maize genes were monitored during the host-pathogen interaction with a custom designed Affymetrix GeneChip® DNA array. More than 4000 maize genes were found differentially expressed at a FDR of 0.05. This included the up regulation of defense related genes and signaling pathways. Transcriptional changes also were observed in primary metabolism genes. Starch biosynthetic genes were down regulated during infection, while genes encoding maize hydrolytic enzymes, presumably involved in the degradation of host reserves, were up regulated. These data indicate that infection of the maize kernel by A. flavus induced metabolic changes in the kernel, including the production of a defense response, as well as a disruption in kernel development. PMID:25132833

  13. Recombinant expression and inhibition mechanism analysis of pectin methylesterase from Aspergillus flavus.

    PubMed

    Jiang, Xiuping; Jia, Qiulei; Chen, Lei; Chen, Qi; Yang, Qing

    2014-06-01

    Phytopathogenic microorganisms can produce pectin methylesterase (PME) to degrade plant cell walls during plant invasion. This enzyme is thought to be a virulence factor of phytopathogens. In this work, PME from Aspergillus flavus (AFPME) was expressed in Pichia pastoris and an in vitro inhibitor study was performed. The purified AFPME with a yield of 52.2% was resolved as one band with a molecular mass of c. 40 kDa by SDS-PAGE. Optimal activity of the enzyme occurred at a temperature of 55 °C and a pH of 4.8. Epigallocatechin gallate (EGCG) strongly inhibited the activity of recombinant AFPME. The molecular docking analysis indicated that EGCG could form hydrogen bonds and π-π interactions with some amino acid residues in the active site of AFPME. Our studies provide a novel strategy for the control of the plant invasion of A. flavus. PMID:24766423

  14. Nanocapsular dispersion of cinnamaldehyde for enhanced inhibitory activity against aflatoxin production by Aspergillus flavus.

    PubMed

    Li, Hongbo; Shen, Qingshan; Zhou, Wei; Mo, Haizhen; Pan, Daodong; Hu, Liangbin

    2015-01-01

    Cinnamaldehyde (CA) is marginally soluble in water, making it challenging to evenly disperse it in foods, and resulting in lowered anti-A. flavus efficacy. In the present study, nano-dispersed CA (nano-CA) was prepared to increase its aqueous solubility. Free and nano-dispersed CA were compared in terms of their inhibitory activity against fungal growth and aflatoxin production of A. flavus both in Sabouraud Dextrose (SD) culture and in peanut butter. Our results indicated that free CA inhibited the mycelia growth and aflatoxin production of A. flavus with a minimal inhibitory concentration (MIC) value of 1.0 mM, but promoted the aflatoxin production at some concentrations lower than the MIC. Nano-CA had a lower MIC value of 0.8 mM against A. flavus, and also showed improved activity against aflatoxin production without the promotion at lower dose. The solidity of peanut butter had an adverse impact on the antifungal activity of free CA, whereas nano-dispersed CA showed more than 2-fold improved activity against the growth of A. flavus. Free CA still promoted AFB1 production at the concentration of 0.25 mM, whereas nano-CA showed more efficient inhibition of AFB1 production in the butter. PMID:25853318

  15. Identification of novel metabolites from Aspergillus flavus by high resolution and multiple stage mass spectrometry.

    PubMed

    Malysheva, Svetlana V; Arroyo-Manzanares, Natalia; Cary, Jeffrey W; Ehrlich, Kenneth C; Vanden Bussche, Julie; Vanhaecke, Lynn; Bhatnagar, Deepak; Di Mavungu, José Diana; De Saeger, Sarah

    2014-01-01

    The filamentous fungus Aspergillus flavus is one of the most important species in the Aspergillus genus and is distributed worldwide as a prevalent aflatoxin-producing food and feed contaminant. A. flavus contains more than 55 gene clusters that are predicted to encode proteins involved in secondary metabolite production. One of these, cluster 27, contains a polyketide synthase (pks27) gene that encodes a protein that is highly homologous to the aflatoxin cluster PKS. Comparative metabolomics, using ultra-high performance liquid chromatography (UHPLC) coupled to high resolution Orbitrap mass spectrometry (MS) was used to detect metabolites differentially expressed in the A. flavus wild-type and ∆pks27 mutant strains. Metabolite profiling was aided by a statistical differential analysis of MS data using SIEVE software. This differential analysis combined with accurate mass data from the Orbitrap and ion trap multiple stage MS allowed four metabolites to be identified that were produced only by the wild-type culture. These included asparasone A (358 Da), an anthraquinone pigment, and related anthraquinones with masses of 316, 340 and 374 Da. These latter three compounds had similar fragmentation patterns to that of asparasone A. The 316 Da anthraquinone is particularly interesting because it is most likely formed by incorporation of seven malonyl-CoA units rather than the eight units required for the formation of asparasone A. The 340 and 374 Da metabolites are the dehydration and an oxy-derivative of asparasone A, respectively. Asparasone A was also identified in extracts from several other Aspergillus species. PMID:24405210

  16. Non-aflatoxigenic Aspergillus flavus to prevent aflatoxin contamination in crops: advantages and limitations

    PubMed Central

    Ehrlich, Kenneth C.

    2014-01-01

    Aspergillus flavus is a diverse assemblage of strains that include aflatoxin-producing and non-toxigenic strains with cosmopolitan distribution. The most promising strategy currently being used to reduce preharvest contamination of crops with aflatoxin is to introduce non-aflatoxin (biocontrol) A. flavus into the crop environment. Whether or not introduction of biocontrol strains into agricultural fields is enough to reduce aflatoxin contamination to levels required for acceptance of the contaminated food as fit for consumption is still unknown. There is no question that biocontrol strains are able to reduce the size of the populations of aflatoxin-producing strains but the available data suggests that at most only a four- to five-fold reduction in aflatoxin contamination is achieved. There are many challenges facing this strategy that are both short term and long term. First, the population biology of A. flavus is not well understood due in part to A. flavus’s diversity, its ability to form heterokaryotic reproductive forms, and its unknown ability to survive for prolonged periods after application. Second, biocontrol strains must be selected that are suitable for the environment, the type of crop, and the soil into which they will be introduced. Third, there is a need to guard against inadvertent introduction of A. flavus strains that could impose an additional burden on food safety and food quality, and fourth, with global warming and resultant changes in the soil nutrients and concomitant microbiome populations, the biocontrol strategy must be sufficiently flexible to adapt to such changes. Understanding genetic variation within strains of A. flavus is important for developing a robust biocontrol strategy and it is unlikely that a “one size fits all” strategy will work for preharvest aflatoxin reduction. PMID:24575088

  17. New Monomeric Stilbenoids from Peanut (Arachis hypogaea) Seeds Challenged by an Aspergillus flavus Strain.

    PubMed

    Sobolev, Victor S; Krausert, Nicole M; Gloer, James B

    2016-01-27

    Two new stilbene derivatives have been isolated from peanut seeds challenged by an Aspergillus flavus strain, along with chiricanine B, which has not been previously reported from peanuts, as well as a stilbenoid reported previously only as a synthetic product. The structures of these new putative phytoalexins were determined by analysis of (1)H and (13)C NMR, HRESIMS, MS(n), and UV data. The new stilbenoids were named arahypin-13 (21), arahypin-14 (22), and arahypin-15 (23). Together with other known bioactive peanut stilbenoids that were also produced in the challenged seeds, these new compounds may play a defensive role against invasive fungi. PMID:26672388

  18. Novel regulation of aflatoxin B1 biosynthesis in Aspergillus flavus by piperonal.

    PubMed

    Park, Eun-Sil; Bae, In Kyung; Kim, Ho Jin; Lee, Sung-Eun

    2016-08-01

    The present study investigated its inhibitory role in aflatoxin (AF) biosynthesis. Treating only AFB1- and B2-producing Aspergillus flavus with piperonal completely inhibited AFB1 production with high sclerotial formation, resulting in 20-fold higher AFG2 production. On the other hand, benzodioxole and eugenol suppressed AFB1 production without AFG formation, while methyleugenol showed potent inhibition of AFB1 production with slight production of AFG1. These results indicate that natural products may change aflatoxin biosynthesis, and highlight a novel regulation of AFG2 production by piperonal. It is the first report for chemical regulation on AFG2 production in non-AFG producing-aspergilli. PMID:26273991

  19. Kojic acid production from cocoa juice by Aspergillus flavus entrapped in calcium alginate.

    PubMed

    el-Sharkawy, S H

    1995-06-01

    Sixteen microorganisms of Aspergillus strains were screened for production of kojic acid using cocoa juice as carbon source. Only Aspergillus flavus ATCC 9179 was found to produce the acid in low yield (22 mg/ml). Calcium alginate immobilization of the cells was used under optimum conditions to maximize the yield of kojic acid (60 mg/ml). Cultures were incubated in the medium with 50% of cocoa juice added in pulses of 8 ml each every 96 hours, and 4% methanol, pH 3.5, 150 rpm, 26 degrees C for three weeks. The incubations were monitored by thin layer and high pressure liquid chromatography. Kojic acid was extracted from the culture broth by organic solvent, concentrated and crystallized. The chemical identity of kojic acid was determined by HPLC, MS, 1H- and 13C-NMR spectroscopy. PMID:7546538

  20. Characterization of Natural Antisense Transcript, Sclerotia Development and Secondary Metabolism by Strand-Specific RNA Sequencing of Aspergillus flavus

    PubMed Central

    Yin, Chao; Guo, Yong; Lin, Ying; Pan, Li; Wang, Bin

    2014-01-01

    Aspergillus flavus has received much attention owing to its severe impact on agriculture and fermented products induced by aflatoxin. Sclerotia morphogenesis is an important process related to A. flavus reproduction and aflatoxin biosynthesis. In order to obtain an extensive transcriptome profile of A. flavus and provide a comprehensive understanding of these physiological processes, the isolated mRNA of A. flavus CA43 cultures was subjected to high-throughput strand-specific RNA sequencing (ssRNA-seq). Our ssRNA-seq data profiled widespread transcription across the A. flavus genome, quantified vast transcripts (73% of total genes) and annotated precise transcript structures, including untranslated regions, upstream open reading frames (ORFs), alternative splicing variants and novel transcripts. We propose natural antisense transcripts in A. flavus might regulate gene expression mainly on the post-transcriptional level. This regulation might be relevant to tune biological processes such as aflatoxin biosynthesis and sclerotia development. Gene Ontology annotation of differentially expressed genes between the mycelia and sclerotia cultures indicated sclerotia development was related closely to A. flavus reproduction. Additionally, we have established the transcriptional profile of aflatoxin biosynthesis and its regulation model. We identified potential genes linking sclerotia development and aflatoxin biosynthesis. These genes could be used as targets for controlled regulation of aflatoxigenic strains of A. flavus. PMID:24849659

  1. Self-sufficient redox biotransformation of lignin-related benzoic acids with Aspergillus flavus.

    PubMed

    Palazzolo, Martín A; Mascotti, María L; Lewkowicz, Elizabeth S; Kurina-Sanz, Marcela

    2015-12-01

    Aromatic carboxylic acids are readily obtained from lignin in biomass processing facilities. However, efficient technologies for lignin valorization are missing. In this work, a microbial screening was conducted to find versatile biocatalysts capable of transforming several benzoic acids structurally related to lignin, employing vanillic acid as model substrate. The wild-type Aspergillus flavus growing cells exhibited exquisite selectivity towards the oxidative decarboxylation product, 2-methoxybenzene-1,4-diol. Interestingly, when assaying a set of structurally related substrates, the biocatalyst displayed the oxidative removal of the carboxyl moiety or its reduction to the primary alcohol whether electron withdrawing or donating groups were present in the aromatic ring, respectively. Additionally, A. flavus proved to be highly tolerant to vanillic acid increasing concentrations (up to 8 g/L), demonstrating its potential application in chemical synthesis. A. flavus growing cells were found to be efficient biotechnological tools to perform self-sufficient, structure-dependent redox reactions. To the best of our knowledge, this is the first report of a biocatalyst exhibiting opposite redox transformations of the carboxylic acid moiety in benzoic acid derivatives, namely oxidative decarboxylation and carboxyl reduction, in a structure-dependent fashion. PMID:26445878

  2. Environmental influences on maize-Aspergillus flavus interactions and aflatoxin production.

    PubMed

    Fountain, Jake C; Scully, Brian T; Ni, Xinzhi; Kemerait, Robert C; Lee, Robert D; Chen, Zhi-Yuan; Guo, Baozhu

    2014-01-01

    Since the early 1960s, the fungal pathogen Aspergillus flavus (Link ex Fr.) has been the focus of intensive research due to the production of carcinogenic and highly toxic secondary metabolites collectively known as aflatoxins following pre-harvest colonization of crops. Given this recurrent problem and the occurrence of a severe aflatoxin outbreak in maize (Zea mays L.), particularly in the Southeast U.S. in the 1977 growing season, a significant research effort has been put forth to determine the nature of the interaction occurring between aflatoxin production, A. flavus, environment and its various hosts before harvest. Many studies have investigated this interaction at the genetic, transcript, and protein levels, and in terms of fungal biology at either pre- or post-harvest time points. Later experiments have indicated that the interaction and overall resistance phenotype of the host is a quantitative trait with a relatively low heritability. In addition, a high degree of environmental interaction has been noted, particularly with sources of abiotic stress for either the host or the fungus such as drought or heat stresses. Here, we review the history of research into this complex interaction and propose future directions for elucidating the relationship between resistance and susceptibility to A. flavus colonization, abiotic stress, and its relationship to oxidative stress in which aflatoxin production may function as a form of antioxidant protection to the producing fungus. PMID:24550905

  3. Utilization of waste fruit-peels to inhibit aflatoxins synthesis by Aspergillus flavus: a biotreatment of rice for safer storage.

    PubMed

    Naseer, R; Sultana, Bushra; Khan, M Z; Naseer, D; Nigam, Poonam

    2014-11-01

    Antifungal activity in lemon and pomegranate peels was considerable against Aspergillus flavus, higher in pomegranate (DIZ 37mm; MIC 135μg/mL). Powdered peels (5, 10, 20% w/w) were mixed in inoculated rice. The inhibitory effect on fungal-growth and production of aflatoxins by A. flavus was investigated at storage conditions - temperature (25, 30°C) and moisture (18%, 21%) for 9months. The maximum total aflatoxins accumulated at 30°C, 21% moisture and at 25°C, 18% moisture were 265.09 and 163.45ng/g, respectively in control. Addition of pomegranate-peels inhibited aflatoxins production to 100% during four month-storage of rice at 25°C and 18% moisture, while lemon-peels showed similar inhibitory effect for 3months at same conditions. However a linear correlation was observed in aflatoxins level with temperature and moisture. Studies showed that both fruit-wastes are potent preventer of aflatoxin production in rice, useful for a safer and longer storage of rice. PMID:25270080

  4. Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris

    PubMed Central

    Karim, Kazi Muhammad Rezaul; Hossain, Md. Anowar; Sing, Ngieng Ngui; Mohd Sinang, Fazia; Hussain, Mohd Hasnain Md.; Roslan, Hairul Azman

    2016-01-01

    A novel thermostable glucoamylase cDNA without starch binding domain (SBD) of Aspergillus flavus NSH9 was successfully identified, isolated, and overexpressed in Pichia pastoris GS115. The complete open reading frame of glucoamylase from Aspergillus flavus NSH9 was identified by employing PCR that encodes 493 amino acids lacking in the SBD. The first 17 amino acids were presumed to be a signal peptide. The cDNA was cloned into Pichia pastoris and the highest expression of recombinant glucoamylase (rGA) was observed after 8 days of incubation period with 1% methanol. The molecular weight of the purified rGA was about 78 kDa and exhibited optimum catalytic activity at pH 5.0 and temperature of 70°C. The enzyme was stable at higher temperature with 50% of residual activity observed after 20 min at 90°C and 100°C. Low concentration of metal (Mg++, Fe++, Zn++, Cu++, and Pb++) had positive effect on rGA activity. This rGA has the potential for use and application in the saccharification steps, due to its thermostability, in the starch processing industries. PMID:27504454

  5. Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris.

    PubMed

    Karim, Kazi Muhammad Rezaul; Husaini, Ahmad; Hossain, Md Anowar; Sing, Ngieng Ngui; Mohd Sinang, Fazia; Hussain, Mohd Hasnain Md; Roslan, Hairul Azman

    2016-01-01

    A novel thermostable glucoamylase cDNA without starch binding domain (SBD) of Aspergillus flavus NSH9 was successfully identified, isolated, and overexpressed in Pichia pastoris GS115. The complete open reading frame of glucoamylase from Aspergillus flavus NSH9 was identified by employing PCR that encodes 493 amino acids lacking in the SBD. The first 17 amino acids were presumed to be a signal peptide. The cDNA was cloned into Pichia pastoris and the highest expression of recombinant glucoamylase (rGA) was observed after 8 days of incubation period with 1% methanol. The molecular weight of the purified rGA was about 78 kDa and exhibited optimum catalytic activity at pH 5.0 and temperature of 70°C. The enzyme was stable at higher temperature with 50% of residual activity observed after 20 min at 90°C and 100°C. Low concentration of metal (Mg(++), Fe(++), Zn(++), Cu(++), and Pb(++)) had positive effect on rGA activity. This rGA has the potential for use and application in the saccharification steps, due to its thermostability, in the starch processing industries. PMID:27504454

  6. Morphological and molecular identification of filamentous Aspergillus flavus and Aspergillus parasiticus isolated from compound feeds in South Africa.

    PubMed

    Iheanacho, Henry E; Njobeh, Patrick B; Dutton, Francis M; Steenkamp, Paul A; Steenkamp, Lucia; Mthombeni, Julian Q; Daru, Barnabas H; Makun, Anthony H

    2014-12-01

    Isolation of filamentous species of two Aspergillum genera from compound feeds produced in South Africa, and subsequent extraction of their individual DNA in this study, presents a simple but rapid molecular procedure for high through-put analysis of the individual morphological forms. DNA was successfully isolated from the Aspergillus spp. from agar cultures by use of a commercial kit. Agarose gel electrophoresis fractionation of the fungi DNA, showed distinct bands. The DNA extracted by this procedure appears to be relatively pure with a ratio absorbance at 260 and 280 nm. However, the overall morphological and molecular data indicated that 67.5 and 51.1% of feed samples were found to be contaminated with Aspergillus flavus and Aspergillus parasiticus, respectively, with poultry feed having the highest contamination mean level of 5.7 × 105 CFU/g when compared to cattle (mean: 4.0 × 106 CFU/g), pig (mean: 2.7 × 104 CFU/g) and horse (1.0 × 102 CFU) feed. This technique presents a readily achievable, easy to use method in the extraction of filamentous fungal DNA and it's identification. Hence serves as an important tool towards molecular study of these organisms for routine analysis check in monitoring and improving compound feed quality against fungal contamination. PMID:25084661

  7. Efficacy of a coating composed of chitosan from Mucor circinelloides and carvacrol to control Aspergillus flavus and the quality of cherry tomato fruits.

    PubMed

    de Souza, Evandro L; Sales, Camila V; de Oliveira, Carlos E V; Lopes, Laênia A A; da Conceição, Maria L; Berger, Lúcia R R; Stamford, Thayza C M

    2015-01-01

    Cherry tomato (Lycopersicon esculentum Mill) fruits are susceptible to contamination by Aspergillus flavus, which may cause the development of fruit rot and significant postharvest losses. Currently there are significant drawbacks for the use of synthetic fungicides to control pathogenic fungi in tomato fruits, and it has increased the interest in exploring new alternatives to control the occurrence of fungal infections in these fruits. This study evaluated the efficacy of chitosan (CHI) from Mucor circinelloides in combination with carvacrol (CAR) in inhibiting A. flavus in laboratory media and as a coating on cherry tomato fruits (25°C, 12 days and 12°C, 24 days). During a period of storage, the effect of coatings composed of CHI and CAR on autochthonous microflora, as well as on some quality characteristics of the fruits such as weight loss, color, firmness, soluble solids, and titratable acidity was evaluated. CHI and CAR displayed MIC valuesof 7.5 mg/mL and 10 μL/mL, respectively, against A. flavus. The combined application of CHI (7.5 or 3.75 mg/mL) and CAR (5 or 2.5 μL/mL) strongly inhibited the mycelial growth and spore germination of A. flavus. The coating composed of CHI (3.75 mg/mL) and CAR (2.5 or 1.25 μL/mL) inhibited the growth of A. flavus in artificially contaminated fruits, as well as the native fungal microflora of the fruits stored at room or low temperature. The application of the tested coatings preserved the quality of cherry tomato fruits as measured by some physicochemical attributes. From this, composite coatings containing CHI and CAR offer a promising alternative to control postharvest infection caused by A. flavus or native fungal microflora in fresh cherry tomato fruits without negatively affecting their quality over storage. PMID:26257717

  8. Efficacy of a coating composed of chitosan from Mucor circinelloides and carvacrol to control Aspergillus flavus and the quality of cherry tomato fruits

    PubMed Central

    de Souza, Evandro L.; Sales, Camila V.; de Oliveira, Carlos E. V.; Lopes, Laênia A. A.; da Conceição, Maria L.; Berger, Lúcia R. R.; Stamford, Thayza C. M.

    2015-01-01

    Cherry tomato (Lycopersicon esculentum Mill) fruits are susceptible to contamination by Aspergillus flavus, which may cause the development of fruit rot and significant postharvest losses. Currently there are significant drawbacks for the use of synthetic fungicides to control pathogenic fungi in tomato fruits, and it has increased the interest in exploring new alternatives to control the occurrence of fungal infections in these fruits. This study evaluated the efficacy of chitosan (CHI) from Mucor circinelloides in combination with carvacrol (CAR) in inhibiting A. flavus in laboratory media and as a coating on cherry tomato fruits (25°C, 12 days and 12°C, 24 days). During a period of storage, the effect of coatings composed of CHI and CAR on autochthonous microflora, as well as on some quality characteristics of the fruits such as weight loss, color, firmness, soluble solids, and titratable acidity was evaluated. CHI and CAR displayed MIC valuesof 7.5 mg/mL and 10 μL/mL, respectively, against A. flavus. The combined application of CHI (7.5 or 3.75 mg/mL) and CAR (5 or 2.5 μL/mL) strongly inhibited the mycelial growth and spore germination of A. flavus. The coating composed of CHI (3.75 mg/mL) and CAR (2.5 or 1.25 μL/mL) inhibited the growth of A. flavus in artificially contaminated fruits, as well as the native fungal microflora of the fruits stored at room or low temperature. The application of the tested coatings preserved the quality of cherry tomato fruits as measured by some physicochemical attributes. From this, composite coatings containing CHI and CAR offer a promising alternative to control postharvest infection caused by A. flavus or native fungal microflora in fresh cherry tomato fruits without negatively affecting their quality over storage. PMID:26257717

  9. RmtA, a Putative Arginine Methyltransferase, Regulates Secondary Metabolism and Development in Aspergillus flavus

    PubMed Central

    Satterlee, Timothy; Cary, Jeffrey W.; Calvo, Ana M.

    2016-01-01

    Aspergillus flavus colonizes numerous oil seed crops such as corn, peanuts, treenuts and cotton worldwide, contaminating them with aflatoxin and other harmful potent toxins. In the phylogenetically related model fungus Aspergillus nidulans, the methyltransferase, RmtA, has been described to be involved in epigenetics regulation through histone modification. Epigenetics regulation affects a variety of cellular processes, including morphogenesis and secondary metabolism. Our study shows that deletion of rmtA in A. flavus results in hyperconidiating colonies, indicating that rmtA is a repressor of asexual development in this fungus. The increase in conidiation in the absence of rmtA coincides with greater expression of brlA, abaA, and wetA compared to that in the wild type. Additionally, the rmtA deletion mutant presents a drastic reduction or loss of sclerotial production, while forced expression of this gene increased the ability of this fungus to generate these resistant structures, revealing rmtA as a positive regulator of sclerotial formation. Importantly, rmtA is also required for the production of aflatoxin B1 in A. flavus, affecting the expression of aflJ. Furthermore, biosynthesis of additional metabolites is also controlled by rmtA, indicating a broad regulatory output in the control of secondary metabolism. This study also revealed that rmtA positively regulates the expression of the global regulatory gene veA, which could contribute to mediate the effects of rmtA on development and secondary metabolism in this relevant opportunistic plant pathogen. PMID:27213959

  10. RmtA, a Putative Arginine Methyltransferase, Regulates Secondary Metabolism and Development in Aspergillus flavus.

    PubMed

    Satterlee, Timothy; Cary, Jeffrey W; Calvo, Ana M

    2016-01-01

    Aspergillus flavus colonizes numerous oil seed crops such as corn, peanuts, treenuts and cotton worldwide, contaminating them with aflatoxin and other harmful potent toxins. In the phylogenetically related model fungus Aspergillus nidulans, the methyltransferase, RmtA, has been described to be involved in epigenetics regulation through histone modification. Epigenetics regulation affects a variety of cellular processes, including morphogenesis and secondary metabolism. Our study shows that deletion of rmtA in A. flavus results in hyperconidiating colonies, indicating that rmtA is a repressor of asexual development in this fungus. The increase in conidiation in the absence of rmtA coincides with greater expression of brlA, abaA, and wetA compared to that in the wild type. Additionally, the rmtA deletion mutant presents a drastic reduction or loss of sclerotial production, while forced expression of this gene increased the ability of this fungus to generate these resistant structures, revealing rmtA as a positive regulator of sclerotial formation. Importantly, rmtA is also required for the production of aflatoxin B1 in A. flavus, affecting the expression of aflJ. Furthermore, biosynthesis of additional metabolites is also controlled by rmtA, indicating a broad regulatory output in the control of secondary metabolism. This study also revealed that rmtA positively regulates the expression of the global regulatory gene veA, which could contribute to mediate the effects of rmtA on development and secondary metabolism in this relevant opportunistic plant pathogen. PMID:27213959

  11. Effects of Gamma and Electron Beam Radiation on Brazil Nuts Artificially Inoculated with Aspergillus flavus.

    PubMed

    Assunção, Ednei; Reis, Tatiana Alves; Baquião, Arianne Costa; Corrêa, Benedito

    2015-07-01

    The aim of this study was to evaluate the effects of gamma radiation (GR) and electron beam (EB) on Brazil nut samples contaminated with Aspergillus flavus. Fifty samples were spread with an A. flavus suspension and incubated at 30°C and a relative humidity of 93%. After 15 days of incubation, mycobiota and aflatoxin analysis were performed. The samples were divided into three groups (control, group 1, and group 2) that received radiation doses of 0 kGy (control) and 5 and 10 kGy each of GR and EB (groups 1 and 2). Noninoculated samples were irradiated with the same doses for sensory evaluation. The results showed that after 15 days of incubation, the average water activity of the samples was 0.80. The irradiation with GR and EB at doses of 5 and 10 kGy was able to eliminate A. flavus in Brazil nut samples. Aflatoxin analysis showed that EB doses of 5 and 10 kGy reduced aflatoxin B1 levels by 53.32 and 65.66%, respectively, whereas the same doses of GR reduced the levels of this toxin by 70.61 and 84.15% compared with the level in the control groups. Sensory evaluation demonstrated that the texture and odor of irradiated Brazil nut samples were acceptable. The taste evaluation indicated that 5 kGy of GR was judged acceptable. The results highlight that both irradiation processes (5- and 10-kGy doses) showed efficiency in A. flavus and aflatoxin elimination. GR and EB treatments resulted in some alterations in the sensory attributes of samples with the doses used in this study; however, Brazil nut samples irradiated with 5-kGy GR doses were considered acceptable. PMID:26197295

  12. Ecology of Aspergillus flavus, Regulation of Aflatoxin Production and Management Strategies to Reduce Aflatoxin Contamination of Corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The contamination of corn (maize) by fungi and the accumulation of mycotoxins are a serious agricultural problem for human and animal health. One particular devastating group of mycotoxins, called aflatoxins, has been intensely studied since the 1960s. Studies of Aspergillus flavus, the agricultura...

  13. Evidence for geographic isolation and distinct patterns of recombination in the aflatoxin gene cluster of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic compounds produced by several Aspergillus species that contaminate food crops worldwide. A. flavus is the most common agent of aflatoxin contamination of corn, peanuts, cottonseed, figs and tree nuts in the US. Extensive studies have elucidated the biochemical and regulatory m...

  14. Use of a Granular Bioplastic Formulation for Carrying Conidia of a Non-aflatoxigenic Strain of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous research demonstrated that aflatoxin contamination in corn grown in Mississippi is reduced by field application of wheat grains pre-inoculated with the non-aflatoxigenic Aspergillus flavus strain NRRL 30797. To facilitate field applications of the biocontrol isolate, a series of laboratory ...

  15. Evaluation of recycled bioplastic pellets and a sprayable formulation for application of an Aspergillus flavus biocontrol strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Biocontrol of Aspergillus flavus using inoculated bioplastic granules has been proven to be effective under laboratory and field conditions. In the present study, the use of low-density pellets from recycled bioplastic as a biocontrol strain carrier was evaluated. Applying recycled bioplastic pell...

  16. Identification of maize genes associated with host plant resistance and susceptibility to Aspergillus flavus infection and aflatoxin accumulation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus infection and aflatoxin contamination of maize pose negative impacts in agriculture and health. Commercial maize hybrids are generally susceptible to this fungus. Significant levels of host plant resistance have been observed in certain maize inbred lines. This study was conducted...

  17. Direct genetic evidence to support the presence of sexual recombination within the life cycle of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus contaminates many important crops worldwide and is the major producer of aflatoxins, which are cancer-causing secondary metabolites. In the United States, mycotoxins have been estimated to cause agricultural losses totaling upwards of $1.4 billion annually, with aflatoxin contamin...

  18. Proteomic analysis of the maize rachis: Potential roles of constitutive and induced proteins in resistance to Aspergillus flavus and aflatoxin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infection of the maize (Zea mays L.) with aflatoxigenic fungus Aspergillus flavus and consequent contamination with carcinogenic aflatoxin is a persistent and serious agricultural problem causing disease and significant crop losses worldwide. The rachis (cob) is an important structure of maize ear ...

  19. Community structure of Aspergillus flavus and A. parasiticus in major almond producing areas of California, United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several nut crops including almonds, pistachios, and walnuts can become contaminated with mycotoxins. Of greatest economic significance are aflatoxins, which are mainly produced by members of Aspergillus section Flavi. The distribution of the two sclerotial-size morphotypes of A. flavus (i.e. S and ...

  20. Comparison of the side-needle and knife techniques for inducing Aspergillus flavus infection and aflatoxin accumulation in corn hybrids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin in corn grain is a problem in many areas of the world. Any combination of environmentally stressful or agronomically unfavorable conditions can increase the likelihood of Aspergillus flavus infection and production of aflatoxin in the corn grain. In the absence of a consistent natural A....

  1. Correlation and Classification of Single Kernel Fluorescence Hyperspectral Data with Aflatoxin Concentration in Corn Kernels Inoculated with Aspergillus flavus Spores

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to examine the relationship between fluorescence emissions of corn kernels inoculated with Aspergillus flavus and aflatoxin contamination levels within the kernels. The choice of methodology was based on the principle that many biological materials exhibit fluorescenc...

  2. Evaluation of the expression genes associated with resistance to Aspergillus flavus colonization and aflatoxin production in different maize lines.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are carcinogenic toxic compounds produced by Aspergillus flavus during infection of crops including maize (Zea mays L.). Contamination of maize with aflatoxin is exacerbated by late season drought stress. Previous studies have implicated numerous resistance-associated proteins (RAPs) that...

  3. Characterization of a maize association mapping panel for new sources of Aspergillus flavus and aflatoxin accumulation resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize (Zea mays L.) susceptibility to ear rot and aflatoxin accumulation by Aspergillus flavus (Link:Fr) causes significant economic and human health damage worldwide. Although host plant resistance is an ideal solution to the problem, no commercial varieties display sufficient levels of resistance ...

  4. Use of functional genomics to assess the impact of climate change on Aspergillus flavus and aflatoxin production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is an opportunistic pathogenic fungus that infects several crops of agricultural importance, among them, corn, cotton, and peanuts. Once established as a pathogen the fungus may secrete secondary metabolites commonly known as mycotoxins, that if consumed by humans or animals may r...

  5. Fluorescent viability stains to probe the metabolic status of aflatoxigenic fungus in dual culture of Aspergillus flavus and Pichia anomala

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The metabolic activity of aflatoxigenic fungus, Aspergillus flavus co-cultured with a biocontrol yeast, Pichia anomala was examined using several vital stains. Both the FUN-1 stain and the combined use of DiBAC4(5) with CDFA-AM stains demonstrated that P. anomala inactivated the ATP generating syst...

  6. Effects of laeA deletion on Aspergillus flavus conidial development and hydrophobicity may contribute to loss of aflatoxin production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The laeA gene encodes a nuclear protein that governs production of multiple fungal secondary metabolites. We examined the effects of laeA deletion in an Aspergillus flavus strain. Compared to wild type, expression of genes involved in secondary metabolism, conidiation and hydrophobicity was drastica...

  7. The Stress Response Regulator AflSkn7 Influences Morphological Development, Stress Response, and Pathogenicity in the Fungus Aspergillus flavus.

    PubMed

    Zhang, Feng; Xu, Gaopo; Geng, Longpo; Lu, Xiaoyan; Yang, Kunlong; Yuan, Jun; Nie, Xinyi; Zhuang, Zhenhong; Wang, Shihua

    2016-01-01

    This study focused on AflSkn7, which is a stress response regulator in the aflatoxin-producing Aspergillus flavus. The ΔAflSkn7 mutants exhibited partially defective conidial formation and a complete inability to generate sclerotia, indicating AflSkn7 affects A. flavus asexual and sexual development. The mutants tolerated osmotic stress but were partially susceptible to the effects of cell wall stress. Additionally, the ΔAflSkn7 mutants were especially sensitive to oxidative stress. These observations confirmed that AflSkn7 influences oxidative stress responses rather than osmotic stress responses. Additionally, AflSkn7 was observed to increase aflatoxin biosynthesis and seed infection rates. These results indicate AflSkn7 affects A. flavus morphological development, stress response, aflatoxin production, and pathogenicity. The results of this study may facilitate the development of new methods to manage A. flavus infections. PMID:27399770

  8. Menadione-Induced Oxidative Stress Re-Shapes the Oxylipin Profile of Aspergillus flavus and Its Lifestyle

    PubMed Central

    Zaccaria, Marco; Ludovici, Matteo; Sanzani, Simona Marianna; Ippolito, Antonio; Aiese Cigliano, Riccardo; Sanseverino, Walter; Scarpari, Marzia; Scala, Valeria; Fanelli, Corrado; Reverberi, Massimo

    2015-01-01

    Aspergillus flavus is an efficient producer of mycotoxins, particularly aflatoxin B1, probably the most hepatocarcinogenic naturally-occurring compound. Although the inducing agents of toxin synthesis are not unanimously identified, there is evidence that oxidative stress is one of the main actors in play. In our study, we use menadione, a quinone extensively implemented in studies on ROS response in animal cells, for causing stress to A. flavus. For uncovering the molecular determinants that drive A. flavus in challenging oxidative stress conditions, we have evaluated a wide spectrum of several different parameters, ranging from metabolic (ROS and oxylipin profile) to transcriptional analysis (RNA-seq). There emerges a scenario in which A. flavus activates several metabolic processes under oxidative stress conditions for limiting the ROS-associated detrimental effects, as well as for triggering adaptive and escape strategies. PMID:26512693

  9. The Stress Response Regulator AflSkn7 Influences Morphological Development, Stress Response, and Pathogenicity in the Fungus Aspergillus flavus

    PubMed Central

    Zhang, Feng; Xu, Gaopo; Geng, Longpo; Lu, Xiaoyan; Yang, Kunlong; Yuan, Jun; Nie, Xinyi; Zhuang, Zhenhong; Wang, Shihua

    2016-01-01

    This study focused on AflSkn7, which is a stress response regulator in the aflatoxin-producing Aspergillus flavus. The ΔAflSkn7 mutants exhibited partially defective conidial formation and a complete inability to generate sclerotia, indicating AflSkn7 affects A. flavus asexual and sexual development. The mutants tolerated osmotic stress but were partially susceptible to the effects of cell wall stress. Additionally, the ΔAflSkn7 mutants were especially sensitive to oxidative stress. These observations confirmed that AflSkn7 influences oxidative stress responses rather than osmotic stress responses. Additionally, AflSkn7 was observed to increase aflatoxin biosynthesis and seed infection rates. These results indicate AflSkn7 affects A. flavus morphological development, stress response, aflatoxin production, and pathogenicity. The results of this study may facilitate the development of new methods to manage A. flavus infections. PMID:27399770

  10. Larval Preference and Performance of Amyelois transitella (Navel Orangeworm, Lepidoptera: Pyralidae) in Relation to the Fungus Aspergillus flavus.

    PubMed

    Ampt, Eline A; Bush, Daniel S; Siegel, Joel P; Berenbaum, May R

    2016-02-01

    The navel orangeworm, Amyelois transitella (Walker), is a polyphagous pest of California nut crops and is responsible for extensive losses in the United States. It directly damages crops by feeding and contaminating nuts with frass and webbing and vectors saprophytic fungi that infect crops. The navel orangeworm is commonly associated with Aspergillus species, including the toxigenic Aspergillus flavus, which causes crop loss by producing carcinogens, including aflatoxin B1. This lepidopteran-fungus association is the most economically serious pest complex in Central Valley orchards, and evidence indicates that this relationship is mutualistic. We assessed preference and performance of navel orangeworm larvae associated with A. flavus in behavioral bioassays in which neonates were allowed to orient within arenas to media with or without fungal tissue, and performance bioassays in which larvae were reared with and without A. flavus on potato dextrose agar (PDA) and a semidefined almond PDA diet to evaluate effects on development and pupal weight. Navel orangeworm larvae were attracted to A. flavus and developed faster in its presence, indicating a nutritional benefit to the caterpillars. Larvae reached pupation ∼33% faster on diet containing A. flavus, and pupal weights were ∼18% higher for males and ∼13% higher for females on this diet. Our findings indicate that A. flavus plays an important role in larval orientation and development on infected hosts. The preference-performance relationship between navel orangeworms and Aspergillus flavus is consistent with a facultative mutualism that has broad implications for pest management efforts and basic understanding of Lepidoptera-plant interactions. PMID:26491042

  11. Effects of Hydrogen Peroxide on Different Toxigenic and Atoxigenic Isolates of Aspergillus flavus

    PubMed Central

    Fountain, Jake C.; Scully, Brian T.; Chen, Zhi-Yuan; Gold, Scott E.; Glenn, Anthony E.; Abbas, Hamed K.; Lee, R. Dewey; Kemerait, Robert C.; Guo, Baozhu

    2015-01-01

    Drought stress in the field has been shown to exacerbate aflatoxin contamination of maize and peanut. Drought and heat stress also produce reactive oxygen species (ROS) in plant tissues. Given the potential correlation between ROS and exacerbated aflatoxin production under drought and heat stress, the objectives of this study were to examine the effects of hydrogen peroxide (H2O2)-induced oxidative stress on the growth of different toxigenic (+) and atoxigenic (−) isolates of Aspergillus flavus and to test whether aflatoxin production affects the H2O2 concentrations that the isolates could survive. Ten isolates were tested: NRRL3357 (+), A9 (+), AF13 (+), Tox4 (+), A1 (−), K49 (−), K54A (−), AF36 (−), and Aflaguard (−); and one A. parasiticus isolate, NRRL2999 (+). These isolates were cultured under a H2O2 gradient ranging from 0 to 50 mM in two different media, aflatoxin-conducive yeast extract-sucrose (YES) and non-conducive yeast extract-peptone (YEP). Fungal growth was inhibited at a high H2O2 concentration, but specific isolates grew well at different H2O2 concentrations. Generally the toxigenic isolates tolerated higher concentrations than did atoxigenic isolates. Increasing H2O2 concentrations in the media resulted in elevated aflatoxin production in toxigenic isolates. In YEP media, the higher concentration of peptone (15%) partially inactivated the H2O2 in the media. In the 1% peptone media, YEP did not affect the H2O2 concentrations that the isolates could survive in comparison with YES media, without aflatoxin production. It is interesting to note that the commercial biocontrol isolates, AF36 (−), and Aflaguard (−), survived at higher levels of stress than other atoxigenic isolates, suggesting that this testing method could potentially be of use in the selection of biocontrol isolates. Further studies will be needed to investigate the mechanisms behind the variability among isolates with regard to their degree of oxidative stress

  12. Aspergillus flavus aflatoxin occurrence and expression of aflatoxin biosynthesis genes in soil.

    PubMed

    Accinelli, Cesare; Abbas, H K; Zablotowicz, R M; Wilkinson, J R

    2008-05-01

    The carcinogen aflatoxin B1 (AFB1) produced by Aspergillus flavus is a major food safety concern in crops. However, information on AFB1 occurrence in soil and crop residue is scarce. A series of experiments investigated the occurrence of AFB1 in soil and corn residues and ascertained the ecology of A. flavus in a Dundee silt loam soil. Samples of untilled soil (0-2 cm) and residues were collected in March 2007 from plots previously planted with a corn isoline containing the Bacillus thuringiensis (Bt) endotoxin gene or the parental non-Bt isoline. AFB1 levels were significantly different in various corn residues. The highest AFB1 levels were observed in cobs containing grain, with 145 and 275 ng.g-1 in Bt and non-Bt, respectively (P > or = F = 0.001). Aflatoxin levels averaged 3.3 and 9.6 ng.g-1 in leaves and (or) stalks and cobs without grain, respectively. All soils had AFB1 ranging from 0.6 to 5.5 ng.g-1 with similar levels in plots from Bt and non-Bt corn. Based on cultural methods, soil contained from log10 3.1 to 4.5 A. flavus cfu.g-1 with about 60% of isolates producing aflatoxin. Laboratory experiments demonstrated that AFB1 is rapidly degraded in soil at 28 degrees C (half-life < or = 5 days). The potential of the soil A. flavus to produce aflatoxins was confirmed by molecular methods. Transcription of 5 aflatoxin biosynthesis genes, including aflD, aflG, aflP, aflR, and aflS, were detected by reverse transcription - polymerase chain reaction analysis in soil. Although AFB1 appears to be transient in soils, it is clear that AFB1 is produced in surface soil in the presence of corn residues, as indicated by A. flavus cfu levels, AFB1 detection, and expression of aflatoxin biosynthetic genes. PMID:18449222

  13. Gene Expression Profiling and Identification of Resistance Genes to Aspergillus flavus Infection in Peanut through EST and Microarray Strategies

    PubMed Central

    Guo, Baozhu; Fedorova, Natalie D.; Chen, Xiaoping; Wan, Chun-Hua; Wang, Wei; Nierman, William C.; Bhatnagar, Deepak; Yu, Jiujiang

    2011-01-01

    Aspergillus flavus and A. parasiticus infect peanut seeds and produce aflatoxins, which are associated with various diseases in domestic animals and humans throughout the world. The most cost-effective strategy to minimize aflatoxin contamination involves the development of peanut cultivars that are resistant to fungal infection and/or aflatoxin production. To identify peanut Aspergillus-interactive and peanut Aspergillus-resistance genes, we carried out a large scale peanut Expressed Sequence Tag (EST) project which we used to construct a peanut glass slide oligonucleotide microarray. The fabricated microarray represents over 40% of the protein coding genes in the peanut genome. For expression profiling, resistant and susceptible peanut cultivars were infected with a mixture of Aspergillus flavus and parasiticus spores. The subsequent microarray analysis identified 62 genes in resistant cultivars that were up-expressed in response to Aspergillus infection. In addition, we identified 22 putative Aspergillus-resistance genes that were constitutively up-expressed in the resistant cultivar in comparison to the susceptible cultivar. Some of these genes were homologous to peanut, corn, and soybean genes that were previously shown to confer resistance to fungal infection. This study is a first step towards a comprehensive genome-scale platform for developing Aspergillus-resistant peanut cultivars through targeted marker-assisted breeding and genetic engineering. PMID:22069737

  14. Genome-wide transcriptome analysis of cotton (Gossypium hirsutum L.) identifies candidate gene signatures in response to aflatoxin producing fungus Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic metabolites and potent carcinogen produced from asexual fungi Aspergillus flavus and A. parasiticus. Aflatoxins can contaminate cottonseed under conducive preharvest and postharvest conditions. U.S. federal regulations restrict the use of aflatoxin contaminated cottonseed at >20...

  15. ROLE OF COMPETITION AND ADVERSE CULTURE CONDITIONS IN PREVENTING THE LOSS OF A AFLATOXIN PRODUCTION BY ASPERGILLUS FLAVUS DURING SERIAL TRANSFERS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is genetically unstable when repeatedly transferred in culture. Serial transfers often result in loss of aflatoxin production and in associated morphological changes such as reduced sporulation, proliferation of aerial hyphae and an inability to produce sclerotia. However, degene...

  16. Extracellular biosynthesis and characterization of silver nanoparticles using Aspergillus flavus NJP08: A mechanism perspective

    NASA Astrophysics Data System (ADS)

    Jain, Navin; Bhargava, Arpit; Majumdar, Sonali; Tarafdar, J. C.; Panwar, Jitendra

    2011-02-01

    The present study demonstrates an eco-friendly and low cost protocol for synthesis of silver nanoparticles using the cell-free filtrate of Aspergillus flavus NJP08 when supplied with aqueous silver (Ag+) ions. Identification of the fungal isolate was based on nuclear ribosomal DNA internal transcribed spacer (ITS) identities. Transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS) revealed the formation of spherical metallic silver nanoparticles. The average particle size calculated using Dynamic Light Scattering measurements (DLS) was found to be 17 +/- 5.9 nm. UV-Visible and Fourier transform infrared (FTIR) spectroscopy confirmed the presence of extracellular proteins. SDS-PAGE profiles of the extracellular proteins showed the presence of two intense bands of 32 and 35 kDa, responsible for the synthesis and stability of silver nanoparticles, respectively. A probable mechanism behind the biosynthesis is discussed, which leads to the possibility of using the present protocol in future ``nano-factories''.

  17. Relating significance and relations of differentially expressed genes in response to Aspergillus flavus infection in maize.

    PubMed

    Asters, Matthew C; Williams, W Paul; Perkins, Andy D; Mylroie, J Erik; Windham, Gary L; Shan, Xueyan

    2014-01-01

    Aspergillus flavus is a pathogenic fungus infecting maize and producing aflatoxins that are health hazards to humans and animals. Characterizing host defense mechanism and prioritizing candidate resistance genes are important to the development of resistant maize germplasm. We investigated methods amenable for the analysis of the significance and relations among maize candidate genes based on the empirical gene expression data obtained by RT-qPCR technique from maize inbred lines. We optimized a pipeline of analysis tools chosen from various programs to provide rigorous statistical analysis and state of the art data visualization. A network-based method was also explored to construct the empirical gene expression relational structures. Maize genes at the centers in the network were considered as important candidate genes for maize DNA marker studies. The methods in this research can be used to analyze large RT-qPCR datasets and establish complex empirical gene relational structures across multiple experimental conditions. PMID:24770700

  18. Relating significance and relations of differentially expressed genes in response to Aspergillus flavus infection in maize

    PubMed Central

    Asters, Matthew C.; Williams, W. Paul; Perkins, Andy D.; Mylroie, J. Erik; Windham, Gary L.; Shan, Xueyan

    2014-01-01

    Aspergillus flavus is a pathogenic fungus infecting maize and producing aflatoxins that are health hazards to humans and animals. Characterizing host defense mechanism and prioritizing candidate resistance genes are important to the development of resistant maize germplasm. We investigated methods amenable for the analysis of the significance and relations among maize candidate genes based on the empirical gene expression data obtained by RT-qPCR technique from maize inbred lines. We optimized a pipeline of analysis tools chosen from various programs to provide rigorous statistical analysis and state of the art data visualization. A network-based method was also explored to construct the empirical gene expression relational structures. Maize genes at the centers in the network were considered as important candidate genes for maize DNA marker studies. The methods in this research can be used to analyze large RT-qPCR datasets and establish complex empirical gene relational structures across multiple experimental conditions. PMID:24770700

  19. Immobilization of Keratinase from Aspergillus flavus K-03 for Degradation of Feather Keratin

    PubMed Central

    2005-01-01

    Extracellular keratinase isolated from Aspergillus flavus K-03 was immobilized on calcium alginate. The properties and reaction activities of free and immobilized keratinase with calcium alginate were characterized. The immobilized keratinase showed proteolytic activity against soluble azo-casein and azo-keratin, and insoluble feather keratin. Heat stability and pH tolerance of keratinase were greatly enhanced by immobilization. It also displayed a higher level of heat stability and an increased tolerance toward alkaline pHs compared with free keratinase. During the durability test at 40℃, 48% of the original enzyme activity of the immobilized keratinase was remained after 7 days of incubation. The immobilized keratinase exhibited better stability, thus increasing its potential for use in industrial application. PMID:24049486

  20. Aflatoxin Biosynthesis and Sclerotial Development in Aspergillus flavus and Aspergillus parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are a family of fungal secondary metabolites. They are produced by species in the genus Aspergillus. Within the last decade, significant advances have been made in understanding the biochemistry, genetics, and gene regulation of aflatoxin biosynthesis. Many scientists have used aflatox...

  1. Inactivation of Aspergillus flavus spores in a sealed package by cold plasma streamers

    NASA Astrophysics Data System (ADS)

    Sohbatzadeh, F.; Mirzanejhad, S.; Shokri, H.; Nikpour, M.

    2016-06-01

    The main objective of this study is to investigate the inactivation efficacy of cold streamers in a sealed package on pathogenic fungi Aspergillus flavus ( A. flavus) spores that artificially contaminated pistachio surface. To produce penetrating cold streamers, electric power supply was adapted to deposit adequate power into the package. The plasma streamers were generated by an alternating high voltage with carrier frequency of 12.5 kHz which was suppressed by a modulated pulsed signal at frequency of 110 Hz. The plasma exposition time was varied from 8 to 18 min to show the effect of the plasma treatment on fungal clearance while the electrode and sample remained at room temperature. This proved a positive effect of the cold streamers treatment on fungal clearance. Benefits of deactivation of fungal spores by streamers inside the package include no heating, short treatment time and adaptability to existing processes. Given its ability to ensure the safety and longevity of food products, this technology has great potential for utilization in food packaging and processing industry. In this study, moisture and pH changes of pistachio samples after plasma streamers treatment were also investigated.

  2. Inactivation of Aspergillus flavus spores in a sealed package by cold plasma streamers

    NASA Astrophysics Data System (ADS)

    Sohbatzadeh, F.; Mirzanejhad, S.; Shokri, H.; Nikpour, M.

    2016-01-01

    The main objective of this study is to investigate the inactivation efficacy of cold streamers in a sealed package on pathogenic fungi Aspergillus flavus (A. flavus) spores that artificially contaminated pistachio surface. To produce penetrating cold streamers, electric power supply was adapted to deposit adequate power into the package. The plasma streamers were generated by an alternating high voltage with carrier frequency of 12.5 kHz which was suppressed by a modulated pulsed signal at frequency of 110 Hz. The plasma exposition time was varied from 8 to 18 min to show the effect of the plasma treatment on fungal clearance while the electrode and sample remained at room temperature. This proved a positive effect of the cold streamers treatment on fungal clearance. Benefits of deactivation of fungal spores by streamers inside the package include no heating, short treatment time and adaptability to existing processes. Given its ability to ensure the safety and longevity of food products, this technology has great potential for utilization in food packaging and processing industry. In this study, moisture and pH changes of pistachio samples after plasma streamers treatment were also investigated.

  3. A Novel Y319H Substitution in CYP51C Associated with Azole Resistance in Aspergillus flavus

    PubMed Central

    Paul, R. A.; Rudramurthy, S. M.; Meis, J. F.; Mouton, J. W.

    2015-01-01

    This study aimed to explore any mutation in the CYP51 gene conferring azole resistance in Aspergillus flavus. Two voriconazole-resistant and 45 voriconazole-susceptible isolates were included in the study. Sequence analysis demonstrated a T1025C nucleotide change in CYP51C, resulting in the Y319H amino acid substitution in one resistant isolate. However, the earlier described T788G mutation in CYP51C conferring voriconazole resistance in A. flavus isolates was present in all isolates, irrespective of their susceptibility status. PMID:26248359

  4. A Novel Y319H Substitution in CYP51C Associated with Azole Resistance in Aspergillus flavus.

    PubMed

    Paul, R A; Rudramurthy, S M; Meis, J F; Mouton, J W; Chakrabarti, A

    2015-10-01

    This study aimed to explore any mutation in the CYP51 gene conferring azole resistance in Aspergillus flavus. Two voriconazole-resistant and 45 voriconazole-susceptible isolates were included in the study. Sequence analysis demonstrated a T1025C nucleotide change in CYP51C, resulting in the Y319H amino acid substitution in one resistant isolate. However, the earlier described T788G mutation in CYP51C conferring voriconazole resistance in A. flavus isolates was present in all isolates, irrespective of their susceptibility status. PMID:26248359

  5. Aspergillus flavus genetic diversity of corn fields treated with non-toxigenic strain afla-guard in the southern U.S

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus genetic diversity of corn fields treated with the non-toxigenic strain Afla-Guard (NRRL 21882) was determined for 384 A. flavus isolates from 14 locations within 6 states in the southern U.S. ELISA test has determined low levels of toxigenic strains (only 91 positive). Nearly hal...

  6. Functional characterization of a veA-dependent polyketide synthase gene in Aspergillus flavus necessary for the synthesis of asparasone, a sclerotium-specific pigment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The filamentous fungus, Aspergillus flavus, produces the toxic and carcinogenic, polyketide synthase (PKS)-derived family of secondary metabolites termed aflatoxins. While analysis of the A. flavus genome has identified many other PKSs capable of producing secondary metabolites, to date, only a few ...

  7. Volatile profiles and aflatoxin production by toxigenic and non-toxigenic isolates of Aspergillus flavus grown on sterile and non-sterile cracked corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a saprophytic fungus which can grow on corn and produce aflatoxins which render it unsafe for food and feed consumption. In this study, aflatoxin and non-aflatoxin producing isolates of A. flavus were grown separately on wet (20% water added), sterile or non-sterile cracked co...

  8. Use of UHPLC high resolution Orbitrap mass spectrometry to investigate genes involved in the production of secondary metabolites in Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungus Aspergillus flavus is known for its ability to produce the toxic and carcinogenic aflatoxins in food and feed. While aflatoxins are of most concern, A. flavus is predicted to be capable of producing many more metabolites based on a study of its complete genome sequence. Some of these meta...

  9. Environmental distribution and genetic diversity of vegetative compatibility groups determine biocontrol strategies to mitigate aflatoxin contamination of maize by Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize infected by aflatoxin-producing Aspergillus flavus may become contaminated with aflatoxins and as a result, threaten human health, food security, and farmers’ income in developing countries where maize is a staple. Environmental distribution and genetic diversity of A. flavus can influence the...

  10. Transcriptome analysis of Aspergillus flavus reveals veA-dependent regulation of secondary metabolite gene clusters, including the novel aflavarin cluster

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The global regulatory veA gene governs development and secondary metabolism in numerous fungal species, including Aspergillus flavus. This is especially relevant since A. flavus infects crops of agricultural importance worldwide, contaminating them with potent mycotoxins. The most well-known are afl...

  11. Intraspecific Aflatoxin Inhibition in Aspergillus flavus Is Thigmoregulated, Independent of Vegetative Compatibility Group and Is Strain Dependent

    PubMed Central

    Huang, Changwei; Jha, Archana; Sweany, Rebecca; DeRobertis, Catherine; Damann,, Kenneth E.

    2011-01-01

    Biological control of preharvest aflatoxin contamination by atoxigenic stains of Aspergillus flavus has been demonstrated in several crops. The assumption is that some form of competition suppresses the fungus's ability to infect or produce aflatoxin when challenged. Intraspecific aflatoxin inhibition was demonstrated by others. This work investigates the mechanistic basis of that phenomenon. A toxigenic and atoxigenic isolate of A. flavus which exhibited intraspecific aflatoxin inhibition when grown together in suspended disc culture were not inhibited when grown in a filter insert-plate well system separated by a .4 or 3 µm membrane. Toxigenic and atoxigenic conidial mixtures (50∶50) placed on both sides of these filters restored inhibition. There was ∼50% inhibition when a 12 µm pore size filter was used. Conidial and mycelial diameters were in the 3.5–7.0 µm range and could pass through the 12 µm filter. Larger pore sizes in the initially separated system restored aflatoxin inhibition. This suggests isolates must come into physical contact with one another. This negates a role for nutrient competition or for soluble diffusible signals or antibiotics in aflatoxin inhibition. The toxigenic isolate was maximally sensitive to inhibition during the first 24 hrs of growth while the atoxigenic isolate was always inhibition competent. The atoxigenic isolate when grown with a green fluorescent protein (GFP) toxigenic isolate failed to inhibit aflatoxin indicating that there is specificity in the touch inhibiton. Several atoxigenic isolates were found which inhibited the GFP isolate. These results suggest that an unknown signaling pathway is initiated in the toxigenic isolate by physical interaction with an appropriate atoxigenic isolate in the first 24 hrs which prevents or down-regulates normal expression of aflatoxin after 3–5 days growth. We suspect thigmo-downregulation of aflatoxin synthesis is the mechanistic basis of intraspecific aflatoxin inhibition

  12. Cloning and functional expression of the gene encoding an inhibitor against Aspergillus flavus alpha-amylase, a novel seed lectin from Lablab purpureus (Dolichos lablab).

    PubMed

    Kim, Young-Hwa; Woloshuk, Charles P; Cho, Eun Hee; Bae, Jung Myung; Song, Young-Sun; Huh, Gyung Hye

    2007-04-01

    Maize is one of the more important agricultural crops in the world and, under certain conditions, prone to attack from pathogenic fungi. One of these, Aspergillus flavus, produces toxic and carcinogenic metabolites, called aflatoxins, as byproducts of its infection of maize kernels. The alpha-amylase of A. flavus is known to promote aflatoxin production in the endosperm of these infected kernels, and a 36-kDa protein from the Lablab purpureus, denoted AILP, has been shown to inhibit alpha-amylase production and the growth of A. flavus. Here, we report the isolation of six full-length labAI genes encoding AILP and a detailed analysis of the activities of the encoded proteins. Each of the six labAI genes encoded sequences of 274 amino acids, with the deduced amino acid sequences showing approximately 95-99% identity. The sequences are similar to those of lectin members of a legume lectin-arcelin-alpha-amylase inhibitor family reported to function in plant resistance to insect pests. The labAI genes did not show any of the structures characteristic of conserved structures identified in alpha-amylase inhibitors to date. The recombinant proteins of labAI-1 and labAI-2 agglutinated human red blood cells and inhibited A. flavus alpha-amylase in a manner similar to that shown by AILP. These data indicate that labAI genes are a new class of lectin members in legume seeds and that their proteins have both lectin and alpha-amylase inhibitor activity. These results are a valuable contribution to our knowledge of plant-pathogen interactions and will be applicable for developing protocols aimed at controlling A. flavus infection. PMID:17149640

  13. Biosynthesis of extracellular and intracellular gold nanoparticles by Aspergillus fumigatus and A. flavus.

    PubMed

    Gupta, Saurabh; Bector, Shruti

    2013-05-01

    Green chemistry is a boon for the development of safe, stable and ecofriendly nanostructures using biological tools. The present study was carried out to explore the potential of selected fungal strains for biosynthesis of intra- and extracellular gold nanostructures. Out of the seven cultures, two fungal strains (SBS-3 and SBS-7) were selected on the basis of development of dark pink colour in cell free supernatant and fungal beads, respectively indicative of extra- and intracellular gold nanoparticles production. Both biomass associated and cell free gold nanoparticles were characterized using X-ray diffractogram (XRD) analysis and transmission electron microscopy (TEM). XRD analysis confirmed crystalline, face-centered cubic lattice of metallic gold nanoparticles along with average crystallite size. A marginal difference in average crystallite size of extracellular (17.76 nm) and intracellular (26 and 22 nm) Au-nanostructures was observed using Scherrer equation. In TEM, a variety of shapes (triangles, spherical, hexagonal) were observed in both extra- and intracellular nanoparticles. 18S rRNA gene sequence analysis by multiple sequence alignment (BLAST) indicated 99 % homology of SBS-3 to Aspergillus fumigatus with 99 % alignment coverage and 98 % homology of SBS-7 to Aspergillus flavus with 98 % alignment coverage respectively. Native-PAGE and activity staining further confirmed enzyme linked synthesis of gold nanoparticles. PMID:23400423

  14. A volatile relationship: profiling an inter-kingdom dialogue between two plant pathogens, Ralstonia Solanacearum and Aspergillus Flavus.

    PubMed

    Spraker, Joseph E; Jewell, Kelsea; Roze, Ludmila V; Scherf, Jacob; Ndagano, Dora; Beaudry, Randolph; Linz, John E; Allen, Caitilyn; Keller, Nancy P

    2014-05-01

    Microbes in the rhizosphere have a suite of extracellular compounds, both primary and secondary, that communicate with other organisms in their immediate environment. Here, we describe a two-way volatile interaction between two widespread and economically important soil-borne pathogens of peanut, Aspergillus flavus and Ralstonia solanacearum, a fungus and bacterium, respectively. In response to A. flavus volatiles, R. solanacearum reduced production of the major virulence factor extracellular polysaccharide (EPS). In parallel, A. flavus responded to R. solanacearum volatiles by reducing conidia production, both on plates and on peanut seeds and by increasing aflatoxin production on peanut. Volatile profiling of these organisms using solid-phase micro-extraction gas chromatography mass spectroscopy (SPME-GCMS) provided a first glimpse at the compounds that may drive these interactions. PMID:24801606

  15. Comparison of expression of secondary metabolite biosynthesis cluster genes in Aspergillus flavus, A. parasiticus, and A. oryzae.

    PubMed

    Ehrlich, Kenneth C; Mack, Brian M

    2014-06-01

    Fifty six secondary metabolite biosynthesis gene clusters are predicted to be in the Aspergillus flavus genome. In spite of this, the biosyntheses of only seven metabolites, including the aflatoxins, kojic acid, cyclopiazonic acid and aflatrem, have been assigned to a particular gene cluster. We used RNA-seq to compare expression of secondary metabolite genes in gene clusters for the closely related fungi A. parasiticus, A. oryzae, and A. flavus S and L sclerotial morphotypes. The data help to refine the identification of probable functional gene clusters within these species. Our results suggest that A. flavus, a prevalent contaminant of maize, cottonseed, peanuts and tree nuts, is capable of producing metabolites which, besides aflatoxin, could be an underappreciated contributor to its toxicity. PMID:24960201

  16. VeA Is Associated with the Response to Oxidative Stress in the Aflatoxin Producer Aspergillus flavus

    PubMed Central

    Baidya, Sachin; Duran, Rocio M.; Lohmar, Jessica M.; Harris-Coward, Pamela Y.; Cary, Jeffrey W.; Hong, Sung-Yong; Roze, Ludmila V.; Linz, John E.

    2014-01-01

    Survival of fungal species depends on the ability of these organisms to respond to environmental stresses. Osmotic stress or high levels of reactive oxygen species (ROS) can cause stress in fungi resulting in growth inhibition. Both eukaryotic and prokaryotic cells have developed numerous mechanisms to counteract and survive the stress in the presence of ROS. In many fungi, the HOG signaling pathway is crucial for the oxidative stress response as well as for osmotic stress response. This study revealed that while the osmotic stress response is only slightly affected by the master regulator veA, this gene, also known to control morphological development and secondary metabolism in numerous fungal species, has a profound effect on the oxidative stress response in the aflatoxin-producing fungus Aspergillus flavus. We found that the expression of A. flavus homolog genes involved in the HOG signaling pathway is regulated by veA. Deletion of veA resulted in a reduction in transcription levels of oxidative stress response genes after exposure to hydrogen peroxide. Furthermore, analyses of the effect of VeA on the promoters of cat1 and trxB indicate that the presence of VeA alters DNA-protein complex formation. This is particularly notable in the cat1 promoter, where the absence of VeA results in abnormally stronger complex formation with reduced cat1 expression and more sensitivity to ROS in a veA deletion mutant, suggesting that VeA might prevent binding of negative transcription regulators to the cat1 promoter. Our study also revealed that veA positively influences the expression of the transcription factor gene atfB and that normal formation of DNA-protein complexes in the cat1 promoter is dependent on AtfB. PMID:24951443

  17. Genome-wide analysis of the Zn(II)₂Cys₆ zinc cluster-encoding gene family in Aspergillus flavus.

    PubMed

    Chang, Perng-Kuang; Ehrlich, Kenneth C

    2013-05-01

    Proteins with a Zn(II)₂Cys₆ domain, Cys-X₂-Cys-X₆-Cys-X₅₋₁₂-Cys-X₂-Cys-X₆₋₉-Cys (hereafter, referred to as the C6 domain), form a subclass of zinc finger proteins found exclusively in fungi and yeast. Genome sequence databases of Saccharomyces cerevisiae and Candida albicans have provided an overview of this family of genes. Annotation of this gene family in most fungal genomes is still far from perfect and refined bioinformatic algorithms are urgently needed. Aspergillus flavus is a saprophytic soil fungus that can produce the carcinogenic aflatoxin. It is the second leading causative agent of invasive aspergillosis. The 37-Mb genome of A. flavus is predicted to encode 12,000 proteins. Two and a half percent of the total proteins are estimated to contain the C6 domain, more than twofold greater than those estimated for yeast, which is about 1 %. The variability in the spacing between cysteines, C₃-C₄ and C₅-C₆, in the zinc cluster enables classification of the domains into distinct subgroups, which are also well conserved in Aspergillus nidulans. Sixty-six percent (202/306) of the A. flavus C6 proteins contain a specific transcription factor domain, and 7 % contain a domain of unknown function, DUF3468. Two A. nidulans C6 proteins containing the DUF3468 are involved in asexual conidiation and another two in sexual differentiation. In the anamorphic A. flavus, a homolog of the latter lacks the C6 domain. A. flavus being heterothallic and reproducing mainly through conidiation appears to have lost some components involved in homothallic sexual development. Of the 55 predicted gene clusters thought to be involved in production of secondary metabolites, only about half have a C6-encoding gene in or near the gene clusters. The features revealed by the A. flavus C6 proteins likely are common for other ascomycete fungi. PMID:23563886

  18. Efficacy of Mentha spicata essential oil in suppression of Aspergillus flavus and aflatoxin contamination in chickpea with particular emphasis to mode of antifungal action.

    PubMed

    Kedia, Akash; Dwivedy, Abhishek Kumar; Jha, Dhruva Kumar; Dubey, Nawal Kishore

    2016-05-01

    The present study reports in vivo antifungal and antiaflatoxigenic efficacy of Mentha spicata essential oil (EO) against toxigenic Aspergillus flavus strain LHP(C)-D6 in chickpea food system up to 12 months of storage. In addition, the mode of antifungal action of EO was also determined to understand the mechanism of fungal growth inhibition. The in vivo study with different concentrations of M. spicata EO showed dose-dependent decrease in fungal colony count as well as aflatoxin B1 concentration. The EO caused >50% protection in inoculated sets and >70% protection in uninoculated sets of chickpea food system against A. flavus at 1.0 μL mL(-1) air concentration. However, at the same concentration, EO caused 100% inhibition to aflatoxin B1 production in both sets when analyzed through high-performance liquid chromatography (HPLC). The antifungal target of EO in fumigated cells of A. flavus was found to be the plasma membrane when analyzed through electron microscopic observations and ions leakage test. The EO fumigated chickpea seeds showed 100% seed germination and seedling growth after 12 months of storage. Based on these observations, M. spicata EO can be recommended as plant-based preservative for safe protection of food commodities during storage conditions against fungal and most importantly mycotoxin contaminations. PMID:26338202

  19. Genetic Similarity among One Aspergillus flavus Strain Isolated from a Patient Who Underwent Heart Surgery and Two Environmental Strains Obtained from the Operating Room

    PubMed Central

    Diaz-Guerra, Teresa M.; Mellado, Emilia; Cuenca-Estrella, Manuel; Gaztelurrutia, Lourdes; Navarro, Jose Ignacio Villate; Tudela, Juan L. Rodríguez

    2000-01-01

    We report the simultaneous isolation of one Aspergillus flavus strain from the aortic prosthesis of a heart surgery patient and another two isolates recovered from a dual-reservoir cooler-heater used in the operating room where this patient was operated on. Genetic typing of these three isolates by randomly amplified polymorphic DNA (RAPD) revealed identical genotypes. Eight unrelated control strains of A. flavus had eight different genotypes. These results clearly indicated the nosocomial origin of the A. flavus strain isolated from the patient. We suggest that the RAPD technique is a rapid and reliable tool to ascertain the epidemiology of infections caused by A. flavus. PMID:10835021

  20. Toward elucidation of genetic and functional genetic mechanisms in corn host resistance to Aspergillus flavus infection and aflatoxin contamination.

    PubMed

    Shan, Xueyan; Williams, W Paul

    2014-01-01

    Aflatoxins are carcinogenic mycotoxins produced by some species in the Aspergillus genus, such as A. flavus and A. parasiticus. Contamination of aflatoxins in corn profusely happens at pre-harvest stage when heat and drought field conditions favor A. flavus colonization. Commercial corn hybrids are generally susceptible to A. flavus infection. An ideal strategy for preventing aflatoxin contamination is through the enhancement of corn host resistance to Aspergillus infection and aflatoxin production. Constant efforts have been made by corn breeders to develop resistant corn genotypes. Significantly low levels of aflatoxin accumulation have been determined in certain resistant corn inbred lines. A number of reports of quantitative trait loci have provided compelling evidence supporting the quantitative trait genetic basis of corn host resistance to aflatoxin accumulation. Important findings have also been obtained from the investigation on candidate resistance genes through transcriptomics approach. Elucidation of molecular mechanisms will provide in-depth understanding of the host-pathogen interactions and hence facilitate the breeding of corn with resistance to A. flavus infection and aflatoxin accumulation. PMID:25101068

  1. Toward elucidation of genetic and functional genetic mechanisms in corn host resistance to Aspergillus flavus infection and aflatoxin contamination

    PubMed Central

    Shan, Xueyan; Williams, W. Paul

    2014-01-01

    Aflatoxins are carcinogenic mycotoxins produced by some species in the Aspergillus genus, such as A. flavus and A. parasiticus. Contamination of aflatoxins in corn profusely happens at pre-harvest stage when heat and drought field conditions favor A. flavus colonization. Commercial corn hybrids are generally susceptible to A. flavus infection. An ideal strategy for preventing aflatoxin contamination is through the enhancement of corn host resistance to Aspergillus infection and aflatoxin production. Constant efforts have been made by corn breeders to develop resistant corn genotypes. Significantly low levels of aflatoxin accumulation have been determined in certain resistant corn inbred lines. A number of reports of quantitative trait loci have provided compelling evidence supporting the quantitative trait genetic basis of corn host resistance to aflatoxin accumulation. Important findings have also been obtained from the investigation on candidate resistance genes through transcriptomics approach. Elucidation of molecular mechanisms will provide in-depth understanding of the host–pathogen interactions and hence facilitate the breeding of corn with resistance to A. flavus infection and aflatoxin accumulation. PMID:25101068

  2. Development and evaluation of ITS- and aflP-based LAMP assays for rapid detection of Aspergillus flavus in food samples.

    PubMed

    Liu, Peiqing; Li, Benjin; Yin, Rongmei; Weng, Qiyong; Chen, Qinghe

    2014-09-01

    Aspergillus flavus is a common filamentous fungus that produces aflatoxins and presents a major threat to agriculture and human health. Previous studies focused mainly on the detection of A. flavus or aflatoxin separately. Here, we developed internal transcribed spacer (ITS)- and aflP-based rapid detection of A. flavus in food samples using the loop-mediated isothermal amplification (LAMP) method. The ITS1-5.8S-ITS2 rDNA region of A. flavus and the aflatoxin-encoding gene aflP were used as target regions. The detection limits of A. flavus and aflP were 10 fg and 1 pg pure DNA, respectively, which allows aflatoxin-contaminated samples to be differentiated from infected samples and reduces false-negative or false-positive results. For specificity testing, DNA extracted from 7 A. flavus, 5 different Aspergillus spp., and 21 other fungi were used, and our results showed that A. flavus strains are detected by ITS-based detection and aflatoxigenic A. flavus strains are detected by aflP-based detection. Furthermore, the ITS- and aflP-based LAMP assays were used for detection analysis of DNA from food samples artificially and naturally contaminated with A. flavus. Our results showed that the detection rate of A. flavus based on the multi-ITS-based LAMP detection is 100% and that the aflatoxigenic strains in all A. flavus are detected by the aflP-based LAMP assay. The LAMP protocol described in our study represents a rapid and highly specific and sensitive diagnostic method for A. flavus detection, which can be used as a diagnostic tool that simplifies A. flavus monitoring and guarantees the quality and safety of foods. PMID:25126831

  3. Effect of corn and peanut cultivation on soil populations of Aspergillus flavus and A. parasiticus in southwestern Georgia.

    PubMed Central

    Horn, B W; Greene, R L; Dorner, J W

    1995-01-01

    The effect of corn and peanut cultivation on the proportion of Aspergillus flavus to A. parasiticus in soil was examined. Soil populations were monitored in three fields during three different years in southwestern Georgia. Each field was planted in both peanuts and corn, and soil was sampled within plots for each crop. A. flavus and A. parasiticus were present in similar proportions in plots from all fields at the beginning of the growing season. A. terreus, A. niger, and A. fumigatus were the other dominant aspergilli in soil. Fields A and B did not show drought stress in peanut or corn plants, and soil populations of A. flavus and A. parasiticus remained stable during the course of the year. In field C, drought stress in corn plants with associated A. flavus infection and aflatoxin contamination greatly increased soil populations of A. flavus relative to A. parasiticus upon dispersal of corn debris to the soil surface by a combine harvester. Colonization of organic debris after it has been added to the soil may maintain soil populations of A. parasiticus despite lower crop infection. PMID:7618858

  4. ROS Involves the Fungicidal Actions of Thymol against Spores of Aspergillus flavus via the Induction of Nitric Oxide

    PubMed Central

    Shen, Qingshan; Zhou, Wei; Li, Hongbo; Hu, Liangbin; Mo, Haizhen

    2016-01-01

    Aspergillus flavus is a well-known pathogenic fungus for both crops and human beings. The acquisition of resistance to azoles by A. flavus is leading to more failures occurring in the prevention of infection by A. flavus. In this study, we found that thymol, one of the major chemical constituents of the essential oil of Monarda punctate, had efficient fungicidal activity against A. flavus and led to sporular lysis. Further studies indicated that thymol treatment induced the generation of both ROS and NO in spores, whereas NO accumulation was far later than ROS accumulation in response to thymol. By blocking ROS production with the inhibitors of NADPH oxidase, NO generation was also significantly inhibited in the presence of thymol, which indicated that ROS induced NO generation in A. flavus in response to thymol treatment. Moreover, the removal of either ROS or NO attenuated lysis and death of spores exposed to thymol. The addition of SNP (exogenous NO donor) eliminated the protective effects of the inhibitors of NADPH oxidase on thymol-induced lysis and death of spores. Taken together, it could be concluded that ROS is involved in spore death induced by thymol via the induction of NO. PMID:27196096

  5. ROS Involves the Fungicidal Actions of Thymol against Spores of Aspergillus flavus via the Induction of Nitric Oxide.

    PubMed

    Shen, Qingshan; Zhou, Wei; Li, Hongbo; Hu, Liangbin; Mo, Haizhen

    2016-01-01

    Aspergillus flavus is a well-known pathogenic fungus for both crops and human beings. The acquisition of resistance to azoles by A. flavus is leading to more failures occurring in the prevention of infection by A. flavus. In this study, we found that thymol, one of the major chemical constituents of the essential oil of Monarda punctate, had efficient fungicidal activity against A. flavus and led to sporular lysis. Further studies indicated that thymol treatment induced the generation of both ROS and NO in spores, whereas NO accumulation was far later than ROS accumulation in response to thymol. By blocking ROS production with the inhibitors of NADPH oxidase, NO generation was also significantly inhibited in the presence of thymol, which indicated that ROS induced NO generation in A. flavus in response to thymol treatment. Moreover, the removal of either ROS or NO attenuated lysis and death of spores exposed to thymol. The addition of SNP (exogenous NO donor) eliminated the protective effects of the inhibitors of NADPH oxidase on thymol-induced lysis and death of spores. Taken together, it could be concluded that ROS is involved in spore death induced by thymol via the induction of NO. PMID:27196096

  6. Characterization of Expressed Sequence Tag-Derived Simple Sequence Repeat Markers for Aspergillus flavus: Emphasis on Variability of Isolates from the Southern United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Simple Sequence Repeat (SSR) markers were developed from Aspergillus flavus expressed sequence tag (EST) database to conduct an analysis of genetic relationships of Aspergillus isolates from numerous host species and geographical regions, but primarily from the United States. Twenty-nine primers wer...

  7. Tissue-specific gene expression in maize seeds during colonization by Aspergillus flavus and Fusarium verticillioides.

    PubMed

    Shu, Xiaomei; Livingston, David P; Franks, Robert G; Boston, Rebecca S; Woloshuk, Charles P; Payne, Gary A

    2015-09-01

    Aspergillus flavus and Fusarium verticillioides are fungal pathogens that colonize maize kernels and produce the harmful mycotoxins aflatoxin and fumonisin, respectively. Management practice based on potential host resistance to reduce contamination by these mycotoxins has proven difficult, resulting in the need for a better understanding of the infection process by these fungi and the response of maize seeds to infection. In this study, we followed the colonization of seeds by histological methods and the transcriptional changes of two maize defence-related genes in specific seed tissues by RNA in situ hybridization. Maize kernels were inoculated with either A. flavus or F. verticillioides 21-22 days after pollination, and harvested at 4, 12, 24, 48, 72, 96 and 120 h post-inoculation. The fungi colonized all tissues of maize seed, but differed in their interactions with aleurone and germ tissues. RNA in situ hybridization showed the induction of the maize pathogenesis-related protein, maize seed (PRms) gene in the aleurone and scutellum on infection by either fungus. Transcripts of the maize sucrose synthase-encoding gene, shrunken-1 (Sh1), were observed in the embryo of non-infected kernels, but were induced on infection by each fungus in the aleurone and scutellum. By comparing histological and RNA in situ hybridization results from adjacent serial sections, we found that the transcripts of these two genes accumulated in tissue prior to the arrival of the advancing pathogens in the seeds. A knowledge of the patterns of colonization and tissue-specific gene expression in response to these fungi will be helpful in the development of resistance. PMID:25469958

  8. Reduction of Aflatoxin in Pistachio Through Biological Control of Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A retrotransposon, AFLAV (A. flavus retrotransposon), has been recently characterized in A. flavus. Complete DNA sequence of 7784 bp containing the AFLAV has been submitted to GenBank (accession number AY485785). Multicopies of this transposon are dispersed in the chromosomes of A. flavus. PCR pr...

  9. Molecular Technique to Fingerprint Aspergillus flavus Causing Aflatoxin Contamination in Food

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A retrotransposon, AFLAV (A. flavus retrotransposon), has been recently characterized in A. flavus. Complete DNA sequence of 7784 bp containing the AFLAV has been submitted to GenBank (accession number AY485785). Multicopies of this transposon are dispersed in the chromosomes of A. flavus. PCR pri...

  10. Sexual Reproduction in Aspergillus flavus Sclerotia: Acquisition of Novel Alleles from Soil Populations and Uniparental Mitochondrial Inheritance

    PubMed Central

    Horn, Bruce W.; Gell, Richard M.; Singh, Rakhi; Sorensen, Ronald B.; Carbone, Ignazio

    2016-01-01

    Aspergillus flavus colonizes agricultural commodities worldwide and contaminates them with carcinogenic aflatoxins. The high genetic diversity of A. flavus populations is largely due to sexual reproduction characterized by the formation of ascospore-bearing ascocarps embedded within sclerotia. A. flavus is heterothallic and laboratory crosses between strains of the opposite mating type produce progeny showing genetic recombination. Sclerotia formed in crops are dispersed onto the soil surface at harvest and are predominantly produced by single strains of one mating type. Less commonly, sclerotia may be fertilized during co-infection of crops with sexually compatible strains. In this study, laboratory and field experiments were performed to examine sexual reproduction in single-strain and fertilized sclerotia following exposure of sclerotia to natural fungal populations in soil. Female and male roles and mitochondrial inheritance in A. flavus were also examined through reciprocal crosses between sclerotia and conidia. Single-strain sclerotia produced ascospores on soil and progeny showed biparental inheritance that included novel alleles originating from fertilization by native soil strains. Sclerotia fertilized in the laboratory and applied to soil before ascocarp formation also produced ascospores with evidence of recombination in progeny, but only known parental alleles were detected. In reciprocal crosses, sclerotia and conidia from both strains functioned as female and male, respectively, indicating A. flavus is hermaphroditic, although the degree of fertility depended upon the parental sources of sclerotia and conidia. All progeny showed maternal inheritance of mitochondria from the sclerotia. Compared to A. flavus populations in crops, soil populations would provide a higher likelihood of exposure of sclerotia to sexually compatible strains and a more diverse source of genetic material for outcrossing. PMID:26731416

  11. Characterization of the Maize Chitinase Genes and Their Effect on Aspergillus flavus and Aflatoxin Accumulation Resistance

    PubMed Central

    Hawkins, Leigh K.; Mylroie, J. Erik; Oliveira, Dafne A.; Smith, J. Spencer; Ozkan, Seval; Windham, Gary L.; Williams, W. Paul; Warburton, Marilyn L.

    2015-01-01

    Maize (Zea mays L.) is a crop of global importance, but prone to contamination by aflatoxins produced by fungi in the genus Aspergillus. The development of resistant germplasm and the identification of genes contributing to resistance would aid in the reduction of the problem with a minimal need for intervention by farmers. Chitinolytic enzymes respond to attack by potential pathogens and have been demonstrated to increase insect and fungal resistance in plants. Here, all chitinase genes in the maize genome were characterized via sequence diversity and expression patterns. Recent evolution within this gene family was noted. Markers from within each gene were developed and used to map the phenotypic effect on resistance of each gene in up to four QTL mapping populations and one association panel. Seven chitinase genes were identified that had alleles associated with increased resistance to aflatoxin accumulation and A. flavus infection in field grown maize. The chitinase in bin 1.05 identified a new and highly significant QTL, while chitinase genes in bins 2.04 and 5.03 fell directly beneath the peaks of previously published QTL. The expression patterns of these genes corroborate possible grain resistance mechanisms. Markers from within the gene sequences or very closely linked to them are presented to aid in the use of marker assisted selection to improve this trait. PMID:26090679

  12. In vitro activities of five antifungal agents against 199 clinical and environmental isolates of Aspergillus flavus, an opportunistic fungal pathogen.

    PubMed

    Khodavaisy, S; Badali, H; Hashemi, S J; Aala, F; Nazeri, M; Nouripour-Sisakht, S; Sorkherizi, M S; Amirizad, K; Aslani, N; Rezaie, S

    2016-06-01

    Aspergillus flavus is the second leading cause of invasive and non-invasive aspergillosis, as well as the most common cause of fungal sinusitis, cutaneous infections, and endophthalmitis in tropical countries. Since resistance to antifungal agents has been observed in patients, susceptibility testing is helpful in defining the activity spectrum of antifungals and determining the appropriate drug for treatment. A collection of 199 clinical and environmental strains of Aspergillus flavus consisted of clinical (n=171) and environmental (n=28) were verified by DNA sequencing of the partial b-tubulin gene. MICs of amphotericin B, itraconazole, voriconazole, posaconazole, and MEC of caspofungin were determined in accordance with the Clinical and Laboratory Standards Institute M38-A2 document. Caspofungin, followed by posaconazole, exhibited the lowest minimum inhibitory concentrations (MIC). All isolates had caspofungin MEC90 (0.063μg/ml) lower than the epidemiologic cutoff values, and 3.5% of the isolates had amphotericin B MIC higher than the epidemiologic cutoff values. However, their clinical effectiveness in the treatment of A. flavus infection remains to be determined. PMID:26948143

  13. Effect of temperature and water activity on gene expression and aflatoxin biosynthesis in Aspergillus flavus on almond medium.

    PubMed

    Gallo, Antonia; Solfrizzo, Michele; Epifani, Filomena; Panzarini, Giuseppe; Perrone, Giancarlo

    2016-01-18

    Almonds are among the commodities at risk of aflatoxin contamination by Aspergillus flavus. Temperature and water activity are the two key determinants in pre and post-harvest environments influencing both the rate of fungal spoilage and aflatoxin production. Varying the combination of these parameters can completely inhibit or fully activate the biosynthesis of aflatoxin, so it is fundamental to know which combinations can control or be conducive to aflatoxin contamination. Little information is available about the influence of these parameters on aflatoxin production on almonds. The objective of this study was to determine the influence of different combinations of temperature (20 °C, 28 °C, and 37 °C) and water activity (0.90, 0.93, 0.96, 0.99 aw) on growth, aflatoxin B1 (AFB1) production and expression of the two regulatory genes, aflR and aflS, and two structural genes, aflD and aflO, of the aflatoxin biosynthetic cluster in A. flavus grown on an almond medium solidified with agar. Maximum accumulation of fungal biomass and AFB1 production was obtained at 28 °C and 0.96 aw; no fungal growth and AFB1 production were observed at 20 °C at the driest tested conditions (0.90 and 0.93 aw). At 20° and 37 °C AFB1 production was 70-90% lower or completely suppressed, depending on aw. Reverse transcriptase quantitative PCR showed that the two regulatory genes (aflR and aflS) were highly expressed at maximum (28 °C) and minimum (20 °C and 37 °C) AFB1 production. Conversely the two structural genes (aflD and aflO) were highly expressed only at maximum AFB1 production (28 °C and 0.96-0.99 aw). It seems that temperature acts as a key factor influencing aflatoxin production which is strictly correlated to the induction of expression of structural biosynthesis genes (aflD and aflO), but not to that of aflatoxin regulatory genes (aflR and aflS), whose functional products are most likely subordinated to other regulatory processes acting at post-translational level

  14. Identification of alternative splice variants in Aspergillus flavus through comparison of multiple tandem MS search algorithms

    PubMed Central

    2011-01-01

    Background Database searching is the most frequently used approach for automated peptide assignment and protein inference of tandem mass spectra. The results, however, depend on the sequences in target databases and on search algorithms. Recently by using an alternative splicing database, we identified more proteins than with the annotated proteins in Aspergillus flavus. In this study, we aimed at finding a greater number of eligible splice variants based on newly available transcript sequences and the latest genome annotation. The improved database was then used to compare four search algorithms: Mascot, OMSSA, X! Tandem, and InsPecT. Results The updated alternative splicing database predicted 15833 putative protein variants, 61% more than the previous results. There was transcript evidence for 50% of the updated genes compared to the previous 35% coverage. Database searches were conducted using the same set of spectral data, search parameters, and protein database but with different algorithms. The false discovery rates of the peptide-spectrum matches were estimated < 2%. The numbers of the total identified proteins varied from 765 to 867 between algorithms. Whereas 42% (1651/3891) of peptide assignments were unanimous, the comparison showed that 51% (568/1114) of the RefSeq proteins and 15% (11/72) of the putative splice variants were inferred by all algorithms. 12 plausible isoforms were discovered by focusing on the consensus peptides which were detected by at least three different algorithms. The analysis found different conserved domains in two putative isoforms of UDP-galactose 4-epimerase. Conclusions We were able to detect dozens of new peptides using the improved alternative splicing database with the recently updated annotation of the A. flavus genome. Unlike the identifications of the peptides and the RefSeq proteins, large variations existed between the putative splice variants identified by different algorithms. 12 candidates of putative isoforms

  15. Efficacy of water-dispersible formulations of biological control strains of Aspergillus flavus for aflatoxin management in corn.

    PubMed

    Weaver, Mark A; Abbas, Hamed K; Jin, Xixuan; Elliott, Brad

    2016-01-01

    Field experiments were conducted in 2011 and 2012 to evaluate the efficacy of water-dispersible granule (WDG) formulations of biocontrol strains of Aspergillus flavus in controlling aflatoxin contamination of corn. In 2011, when aflatoxin was present at very high levels, there was no WDG treatment that could provide significant protection against aflatoxin contamination. The following year a new WDG formulation was tested that resulted in 100% reduction in aflatoxin in one field experiment and ≥ 49% reduction in all five WDG treatments with biocontrol strain 21882. Large sampling error, however, limited the resolution of various treatment effects. Corn samples were also subjected to microbial analysis to understand better the mechanisms of successful biocontrol. In the samples examined here, the size of the A. flavus population on the grain was associated with the amount of aflatoxin, but the toxigenic status of that population was a poor predictor of aflatoxin concentration. PMID:26730792

  16. The Master Transcription Factor mtfA Governs Aflatoxin Production, Morphological Development and Pathogenicity in the Fungus Aspergillus flavus

    PubMed Central

    Zhuang, Zhenhong; Lohmar, Jessica M.; Satterlee, Timothy; Cary, Jeffrey W.; Calvo, Ana M.

    2016-01-01

    Aspergillus flavus produces a variety of toxic secondary metabolites; among them, the aflatoxins (AFs) are the most well known. These compounds are highly mutagenic and carcinogenic, particularly AFB1. A. flavus is capable of colonizing a number of economically-important crops, such as corn, cotton, peanut and tree nuts, and contaminating them with AFs. Molecular genetic studies in A. flavus could identify novel gene targets for use in strategies to reduce AF contamination and its adverse impact on food and feed supplies worldwide. In the current study, we investigated the role of the master transcription factor gene mtfA in A. flavus. Our results revealed that forced overexpression of mtfA results in a drastic decrease or elimination of several secondary metabolites, among them AFB1. The reduction in AFB1 was accompanied by a decrease in aflR expression. Furthermore, mtfA also regulates development; conidiation was influenced differently by this gene depending on the type of colonized substrate. In addition to its effect on conidiation, mtfA is necessary for the normal maturation of sclerotia. Importantly, mtfA positively affects the pathogenicity of A. flavus when colonizing peanut seeds. AF production in colonized seeds was decreased in the deletion mtfA strain and particularly in the overexpression strain, where only trace amounts were detected. Interestingly, a more rapid colonization of the seed tissue occurred when mtfA was overexpressed, coinciding with an increase in lipase activity and faster maceration of the oily part of the seed. PMID:26805883

  17. Efficacy of the combined application of chitosan and Locust Bean Gum with different citrus essential oils to control postharvest spoilage caused by Aspergillus flavus in dates.

    PubMed

    Aloui, Hajer; Khwaldia, Khaoula; Licciardello, Fabio; Mazzaglia, Agata; Muratore, Giuseppe; Hamdi, Moktar; Restuccia, Cristina

    2014-01-17

    This study reports the efficacy of the combined application of chitosan (CH) and Locust Bean Gum (LBG) in combination with different citrus essential oils (EOs) to inhibit Aspergillus flavus in vitro and on artificially infected dates for a storage period of 12 days. The effect of these treatments on the fruits' sensory characteristics was evaluated to verify the complete absence of off-odours and off-flavours. Bergamot EO was the most effective in reducing mycelial growth, followed by bitter orange EO. Both bergamot and bitter orange oils significantly reduced conidial germination and a complete inhibition was obtained at concentrations higher than 2%. The mixtures based on CH-2% (v/v) bergamot EO or CH-2% (v/v) bitter orange EO proved to be the most effective coatings to reduce conidial germination resulting in an 87-90% inhibition compared with the control. In fruit decay assays coatings based on CH incorporating citrus oils were able to reduce fungal decay in the range of 52-62% at day 12. The study results and the complete absence of off-flavours and off-odours demonstrate the potential of CH coatings carrying citrus EOs at sub-inhibitory concentrations to control postharvest growth of A. flavus in dates. PMID:24291176

  18. EVALUATION OF A BIOPESTICIDE, PICHIA ANOMALA WRL-076 TO CONTROL ASPERGILLUS FLAVUS IN A COMMERCIAL ORCHARD

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Existing literatures indicate that wounds in plant tissues provide the entry to A. flavus. By mechanically wounding pistachio nut-fruits, sufficient number of nut-fruits conducive to A. flavus and fungal infection are generated. The wounded nut-fruits are easily recognized for sampling. Two experim...

  19. Purification and Characterization of a Keratinase from a Feather-Degrading Fungus, Aspergillus flavus Strain K-03

    PubMed Central

    2007-01-01

    A keratinolytic enzyme secreted by Aspergillus flavus K-03 cultured in feather meal basal medium (FMBM) containing 2% (w/v) chicken feather was purified and characterized. Keratinolytic enzyme secretion was the maximal at day 16 of the incubation period at pH 8 and 28℃. No relationship was detected between enzyme yield and increase of fungal biomass. The fraction obtained at 80% ammonium sulfate saturation showed 2.39-fold purification and was further purified by gel filtration in Sephadex G-100 followed by ion exchange chromatography on DEAE-Sephadex A-50, yielding an active protein peak showing 11.53-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymograms indicated that the purified keratinase is a monomeric enzyme with 31 kDa molecular weight. The extracellular keratinase of A. flavus was active in a board range of pH (7~10) and temperature (30℃~70℃) profiles with the optimal for keratinase activity at pH 8 and 45℃. The keratinase activity was totally inhibited by protease inhibitors such as phenylmethylsulfonyl fluoride (PMSF), iodoacetic acid, and ethylenediaminetetraacetate (EDTA) while no reduction of activity by the addition of dithiothreitol (DTT) was observed. N-terminal amino acid sequences were up to 80% homologous with the fungal subtilisins produced by Fusarium culmorum. Therefore, on the basis of these characteristics, the keratinase of A. flavus K-03 is determined to be subtilisins-like. PMID:24015101

  20. Overexpression of aflR Leads to Upregulation of Pathway Gene Transcription and Increased Aflatoxin Production in Aspergillus flavus

    PubMed Central

    Flaherty, J. E.; Payne, G. A.

    1997-01-01

    The aflatoxin biosynthetic pathway regulatory gene, aflR, encodes a putative 47-kDa protein containing a zinc cluster DNA binding motif. It is required for the transcription of all of the characterized aflatoxin pathway genes in both Aspergillus flavus and Aspergillus parasiticus. The objective of this study was to examine the effects of aflR overexpression on temporal gene expression, aflatoxin production, and nitrate inhibition of aflatoxin biosynthesis in A. flavus. An inducible expression construct was made by fusing the coding region of aflR to the promoter region of the A. flavus adh1 gene. This construct was transformed into A. flavus 656-2 (FGSC A1010), a strain mutated at the aflR locus. Strain 656-2 containing the adh1(p)::aflR construct had induced transcription of two early aflatoxin pathway genes, nor-1 and pksA, and produced wild-type concentrations of aflatoxin in a temporal pattern similar to that of wild-type strains of A. flavus. Strains 656-2 and 86-10 (FGSC A1009) an aflatoxigenic strain, were transformed with a construct containing the constitutive promoter gpdA driving aflR. Transformants of these strains constitutively expressed aflR, fas-1A, pksA, nor-1, and omtA but did not constitutively produce aflatoxin. Strain 86-10 containing the gpdA(p)::aflR construct produced 50 times more aflatoxin than 86-10, but the temporal pattern of aflatoxin production was the same as for 86-10, and aflatoxin production was also induced by sucrose. The addition of 10 g of nitrate per liter to sucrose low salts medium inhibited aflatoxin production by both strain 86-10 and a transformant of 86-10 containing the gpdA(p)::aflR construct, indicating that nitrate inhibition of aflatoxin biosynthesis does not occur solely at the level of aflR transcription. These studies show that constitutive overexpression of the pathway transcriptional regulatory gene aflR leads to higher transcript accumulation of pathway genes and increased aflatoxin production but that the

  1. Morphological and toxigenic variability in the Aspergillus flavus isolates from peanut (Arachis hypogaea L.) production system in Gujarat (India).

    PubMed

    Singh, Diwakar; Thankappan, Radhakrishnan; Kumar, Vinod; Bagwan, Naimoddin B; Basu, Mukti S; Dobaria, Jentilal R; Mishra, Gyan P; Chanda, Sumitra

    2015-03-01

    Morphological and toxigenic variability in 187 Aspergillus flavus isolates, collected from a major Indian peanut production system, from 10 districts of Gujarat was studied. On the basis of colony characteristics, the isolates were grouped as group A (83%), B (11%) and G (6%). Of all the isolates, 21%, 47% and 32% were found to be fast-growing, moderately-fast and slow-growing respectively, and nosclerotia and sclerotia production was recorded in 32.1% and 67% isolates respectively. Large, medium and small number of sclerotia production was observed in 55, 38 and 34 isolates respectively. Toxigenic potential based on ammonia vapour test was not found reliable, while ELISA test identified 68.5%, 18.7% and 12.8% isolates as atoxigenic, moderately-toxigenic and highly-toxigenic, respectively. On clustering, the isolates were grouped into 15 distinct clusters, 'A' group of isolates was grouped distinctly in different clusters, while 'B' and 'G' groups of isolates were clustered together. No association was observed between morphological-diversity and toxigenic potential of the isolates. From the present investigation, most virulent isolates were pooled to form a consortium for sick-plot screening of germplasm, against Aspergillus flavus. In future, atoxigenic isolates may be evaluated for their potential to be used as bio-control agent against toxigenicisolates. PMID:25895268

  2. Optimization of process parameters influencing the submerged fermentation of extracellular lipases from Pseudomonas aeruginosa, candida albicans and Aspergillus flavus.

    PubMed

    Padhiar, Jigita; Das, Arijit; Bhattacharya, Sourav

    2011-11-15

    The present study was aimed at optimization, production and partial purification of lipases from Pseudomonas aeruginosa, Candida albicans and Aspergillus flavus. Various nutritional and physical parameters affecting lipase production such as carbon and nitrogen supplements, pH, temperature, agitation speed and incubation time were studied. Refined sunflower oil (1% v/v) and tryptone at a pH of 6.2 favored maximum lipase production in Pseudomonas at 30 degrees C and 150 rpm, when incubated for 5 days. In C. albicans refined sunflower oil (3% v/v) and peptone resulted in maximum lipase production at pH 5.2, 30 degrees C and 150 rpm, when incubated for 5 days. In A. flavus coconut oil (3% v/v) and peptone yielded maximum lipase at pH 6.2, 37 degrees C, 200 rpm after an incubation period of 5 days. The lipases were partially purified by ammonium sulphate precipitation and dialysis. In P. aeruginosa enzyme activity of the dialyzed fraction was found to be 400 U mL-' and for C. albicans 410 U mL(-1). The dialysed lipase fraction from A. flavus demonstrated an activity of 460 U mL(-1). The apparent molecular weights of the dialyzed lipases were determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The dialyzed lipase fraction obtained from P. aeruginosa revealed molecular weights of 47, 49 and 51 kDa, whereas, lipases from C. albicans and A. flavus demonstrated 3 bands (16.5, 27 and 51 kDa) and one band (47 kDa), respectively. These extracellular lipases may find wide industrial applications. PMID:22514878

  3. The two genome sequence release and blast server construction for aflatoxin-producing L and S strains Aspergillus parasiticus and A. flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are toxic and carcinogenic secondary metabolites. These compounds, produced by Aspergillus flavus and A. parasiticus, contaminate pre-harvest agricultural crops in the field and post-harvest grains during storage. In order to reduce and eliminate aflatoxin contamination of food and feed...

  4. Field efficacy of a mixture of atoxigenic Aspergillus flavus Link:Fr vegetative compatibility groups in preventing aflatoxin contamination in maize (Zea mays L.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Competitive exclusion of aflatoxin producers by endemic atoxigenic strains of Aspergillus flavus is a proven tool for aflatoxin management being adapted for use in Africa. Field efficacy of an experimental formulation consisting of four native atoxigenic strains (La3303, La3304, La3279 and Ka16127) ...

  5. An Aspergillus flavus secondary metabolic gene cluster containing a hybrid PKS-NRPS is necessary for synthesis of the 2-pyridones, leporins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genome of the filamentous fungus, Aspergillus flavus, has been shown to harbor as many as 55 putative secondary metabolic gene clusters including the one responsible for production of the toxic and carcinogenic, polyketide synthase (PKS)-derived family of secondary metabolites termed aflatoxins....

  6. Postharvest Aspergillus flavus colonization in responding to preharvest field condition of drought stress and oligo-macroarray profiling of developing corn kernel gene expression under drought stress

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Drought stress is a major factor known to contribute to preharvest aflatoxin contamination of maize kernels. Aspergillus flavus infection of maize kernels occurs earlier than aflatoxin accumulation in developing kernels. Recent studies have demonstrated higher concentration of defense or stress-rela...

  7. Genetic analysis of the Aspergillus flavus vegetative compatibility group to which a biological control agent that limits aflatoxin contamination in USA crops belongs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The most effective means of reducing aflatoxin contamination of agricultural crops is through competitive exclusion of aflatoxin-producing fungi using vegetative compatibility groups (VCGs) of Aspergillus flavus that do not produce aflatoxin. Presently there are two VCGs in the USA registered with t...

  8. A public platform for the verification of the phenotypic effect of candidate genes for resistance to aflatoxin accumulation and Aspergillus flavus infection in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A public candidate gene testing pipeline for resistance to aflatoxin accumulation or Aspergillus flavus infection in maize is presented here. The pipeline consists of steps for identifying, testing, and verifying the association of any maize gene sequence with resistance under field conditions. Reso...

  9. Effect of inoculum concentrations of Aspergillus flavus and A. parasiticus on aflatoxin accumulation and kernel infection in resistant and susceptible maize hybrids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Over a three year period, we compared aflatoxin accumulation and kernel infection in maize hybrids inoculated with six inoculum concentrations of Aspergillus flavus isolate NRRL 3357 or A. parasiticus isolate NRRL 6111 which is a norsolorinic acid producer. Aflatoxin resistant and susceptible mai...

  10. Non-pheromonal control of navel orangeworm as a promising method toward decreasing contamination of Aspergillus flavus in California tree nuts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The navel orangeworm (NOW) is a major insect pest of tree nuts and is a vector of Aspergillus flavus – a fungus responsible for aflatoxin contamination of California tree nuts. Despite the presence of NOW throughout a typical season, the identification of particular VOCs, or their potential role as ...

  11. Effect of nontoxigenic Aspergillus flavus and A. parasiticus on aflatoxin contamination of wounded peanut seeds inoculated with agricultural soil containing natural fungal populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanuts and other seed and grain crops are commonly contaminated with carcinogenic aflatoxins, secondary metabolites produced by Aspergillus flavus and A. parasiticus. Aflatoxin contamination of peanuts in the field can be reduced by 77 to 98% with biological control through the application of nont...

  12. RNAi silencing of the 14 kDa trypsin inhibitor protein in maize and its effect on host resistance against Aspergillus flavus infection/aflatoxin production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize (Zea mays L.) is one of the major crops susceptible to Aspergillus flavus Link ex. Fries infection and subsequent aflatoxin contamination. Previous studies found the expression of an antifungal 14 kDa trypsin inhibitor (TI) was associated with maize aflatoxin resistance. To further investigate...

  13. Effect of exogenous jasmonic acid application on Aspergillus flavus kernel infection and aflatoxin production in two maize hybrids (Zea mays L.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Jasmonic acid (JA), produced by the octadecanoid pathway, is a phytohormone that triggers induced resistance against certain pathogens and arthropod herbivores. The octadecanoid pathway has been implicated in playing a role in the Aspergillus flavus-maize seed interaction. In field studies, the ef...

  14. Identification and quantification of a toxigenic and non-toxigenic Aspergillus flavus strain in contaminated maize using quantitative real-time PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins, which are produced by the fungus Aspergillus flavus, are toxic to humans, livestock, and pets. The value of maize (Zea mays) grain is markedly reduced when contaminated with aflatoxin. Plant resistance and biological control using non-toxin producing strains are considered effective st...

  15. Non-aflatoxigenic Aspergillus flavus as potential biocontrol agents to reduce aflatoxin contamination in peanuts harvested in Northern Argentina.

    PubMed

    Alaniz Zanon, María Silvina; Barros, Germán Gustavo; Chulze, Sofía Noemí

    2016-08-16

    Biological control is one of the most promising strategies for preventing aflatoxin contamination in peanuts at field stage. A population of 46 native Aspergillus flavus nonaflatoxin producers were analysed based on phenotypic, physiological and genetic characteristics. Thirty-three isolates were characterized as L strain morphotype, 3 isolates as S strain morphotype, and 10 isolates did not produce sclerotia. Only 11 of 46 non-aflatoxigenic isolates did not produce cyclopiazonic acid. The vegetative compatibility group (VCG) diversity index for the population was 0.37. For field trials we selected the non-aflatoxigenic A. flavus AR27, AR100G and AFCHG2 strains. The efficacy of single and mixed inocula as potential biocontrol agents in Northern Argentina was evaluated through a 2-year study (2014-2015). During the 2014 peanut growing season, most of the treatments reduced the incidence of aflatoxigenic strains in both soil and peanut kernel samples, and no aflatoxin was detected in kernels. During the 2015 growing season, there was a reduction of aflatoxigenic strains in kernel samples from the plots treated with the potential biocontrol agents. Reductions of aflatoxin contamination between 78.36% and 89.55% were observed in treated plots in comparison with the un-inoculated control plots. This study provides the first data on aflatoxin biocontrol based on competitive exclusion in the peanut growing region of Northern Argentina, and proposes bioproducts with potential use as biocontrol agents. PMID:27220011

  16. Transcriptional profiles uncover Aspergillus flavus-induced resistance in maize kernels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination caused by the opportunistic pathogen A. flavus is a major concern in maize production prior to harvest and during storage, and also a concern in many other crops, such as peanuts, cottonseed, tree nuts, and rice. Although a number of resistant maize lines with low aflatoxin c...

  17. Analysis of genetic and aflatoxin diversity among Aspergillus flavus strains isolated from sorghum seeds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A total of 34 A. flavus isolates were recovered from sorghum seeds sampled across five states in India. Our study included (1) species confirmation through PCR assay, (2) an aflatoxin cluster genotype assay using developed multiplex PCR, (3) quantification of total aflatoxin concentrations by the iC...

  18. Characterization of AFLAV, a Tfl/Sushi retrotransposon from Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The insert of pAF28 contains a 4.5 kb region which encodes a truncated retrotransposon (AfRTL-1). In search for a full-length and intact copy of retrotransposon, we exploited a novel PCR cloning strategy by amplifying a 3.4 kb region from the genomic DNA of A. flavus NRRL 6541. The fragment was cl...

  19. rtfA, a putative RNA-Pol II transcription elongation factor gene, is necessary for normal morphological and chemical development in Aspergillus flavus.

    PubMed

    Lohmar, Jessica M; Harris-Coward, Pamela Y; Cary, Jeffrey W; Dhingra, Sourabh; Calvo, Ana M

    2016-06-01

    The filamentous fungus Aspergillus flavus is an agriculturally important opportunistic plant pathogen that produces potent carcinogenic compounds called aflatoxins. We identified the A. flavus rtfA gene, the ortholog of rtf1 in Saccharomyces cerevisiae and rtfA in Aspergillus nidulans. Interestingly, rtfA has multiple cellular roles in this mycotoxin-producing fungus. In this study, we show that rtfA regulates conidiation. The rtfA deletion mutant presented smaller conidiophores with significantly reduced conidial production compared to the wild-type strain. The absence of rtfA also resulted in a significant decrease or lack of sclerotial production under conditions that allowed abundant production of these resistance structures in the wild type. Importantly, the deletion of rtfA notably reduced the production of aflatoxin B1, indicating that rtfA is a regulator of mycotoxin biosynthesis in A. flavus. In addition, the deletion rtfA also altered the production of several unknown secondary metabolites indicating a broader regulatory scope. Furthermore, our study revealed that rtfA controls the expression of the global regulators veA and laeA, which further influence morphogenesis and secondary metabolism in A. flavus. PMID:27020290

  20. Molecular variation analysis of Aspergillus flavus using polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer rDNA region

    PubMed Central

    Zarrin, Majid; Erfaninejad, Maryam

    2016-01-01

    Aspergillus flavus is the second most common disease-causing species of Aspergillus in humans. The fungus is frequently associated with life-threatening infections in immunocompromised hosts. The primary aim of the present study was to analyze the genetic variability among different isolates of A. flavus using polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP). A total of 62 A. flavus isolates were tested in the study. Molecular variability was searched for by analysis of the PCR amplification of the internal transcribed spacer (ITS) regions of ribosomal DNA using restriction enzymes. PCR using primers for ITS1 and ITS4 resulted in a product of ~600 bp. Amplicons were subjected to digestion with restriction endonucleases EcoRI, HaeIII and TaqI. Digestion of the PCR products using these restriction enzymes produced different patterns of fragments among the isolates, with different sizes and numbers of fragments, revealing genetic variability. In conclusion, ITS-RFLP is a useful molecular tool in screening for nucleotide polymorphisms among A. flavus isolates. PMID:27588085

  1. Comparative Analysis of the Performance of Aspergillus flavus on Resistant and Susceptible Maize Genotypes during Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus, a mycotoxicogenic fungal genus, produces carcinogenic aflatoxins in crops like peanuts and maize. Development of fungal resistant maize cultivars is one strategy used to decrease contamination. Successful development and identification of resistant maize genotypes requires evaluation o...

  2. Aspergillus Flavus/Aflatoxin Occurrence and Expression of Aflatoxin Biosynthesis Genes in Soil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycotoxins, including aflatoxins, fumonisins, cyclopiazonic acid (CPA), and zearalenone, produced by Aspergillus and Fusarium species when present in grain can cause serious health problems in livestock and humans. Little is known about the occurrence of these toxins in corn plant debris post-harve...

  3. Aspergillus flavus Conidia-derived Carbon/Sulfur Composite as a Cathode Material for High Performance Lithium–Sulfur Battery

    PubMed Central

    Xu, Maowen; Jia, Min; Mao, Cuiping; Liu, Sangui; Bao, Shujuan; Jiang, Jian; Liu, Yang; Lu, Zhisong

    2016-01-01

    A novel approach was developed to prepare porous carbon materials with an extremely high surface area of 2459.6 m2g−1 by using Aspergillus flavus conidia as precursors. The porous carbon serves as a superior cathode material to anchor sulfur due to its uniform and tortuous morphology, enabling high capacity and good cycle lifetime in lithium sulfur-batteries. Under a current rate of 0.2 C, the carbon-sulfur composites with 56.7 wt% sulfur loading deliver an initial capacity of 1625 mAh g−1, which is almost equal to the theoretical capacity of sulfur. The good performance may be ascribed to excellent electronic networks constructed by the high-surface-area carbon species. Moreover, the semi-closed architecture of derived carbons can effectively retard the polysulfides dissolution during charge/discharge, resulting in a capacity of 940 mAh g−1 after 120 charge/discharge cycles. PMID:26732547

  4. Potential for aflatoxin B1 and B2 production by Aspergillus flavus strains isolated from rice samples

    PubMed Central

    Lai, Xianwen; Zhang, He; Liu, Ruicen; Liu, Chenglan

    2014-01-01

    In this study, we investigated the potential for aflatoxin B1 (AFB1) and B2 (AFB2) production in rice grain by 127 strains of Aspergillus flavus isolated from rice grains collected from China. These strains were inoculated onto rice grains and incubated at 28 °C for 21 days. AFB1 and AFB2 were extracted and quantified by high-performance liquid chromatography coupled with fluorescence detection. Among the tested strains, 37% produced AFB1 and AFB2 with levels ranging from 175 to 124 101 μg kg−1 for AFB1 and from not detected to 10 329 μg kg−1 for AFB2. The mean yields of these isolates were 5884 μg kg−1 for AFB1 and 1968 μg kg−1 for AFB2. Overall, most of the aflatoxigenic strains produced higher levels of AFB1 than AFB2 in rice. The obtained information is useful for assessing the risk of aflatoxin contamination in rice samples. PMID:25737649

  5. Aspergillus flavus Conidia-derived Carbon/Sulfur Composite as a Cathode Material for High Performance Lithium-Sulfur Battery

    NASA Astrophysics Data System (ADS)

    Xu, Maowen; Jia, Min; Mao, Cuiping; Liu, Sangui; Bao, Shujuan; Jiang, Jian; Liu, Yang; Lu, Zhisong

    2016-01-01

    A novel approach was developed to prepare porous carbon materials with an extremely high surface area of 2459.6 m2g-1 by using Aspergillus flavus conidia as precursors. The porous carbon serves as a superior cathode material to anchor sulfur due to its uniform and tortuous morphology, enabling high capacity and good cycle lifetime in lithium sulfur-batteries. Under a current rate of 0.2 C, the carbon-sulfur composites with 56.7 wt% sulfur loading deliver an initial capacity of 1625 mAh g-1, which is almost equal to the theoretical capacity of sulfur. The good performance may be ascribed to excellent electronic networks constructed by the high-surface-area carbon species. Moreover, the semi-closed architecture of derived carbons can effectively retard the polysulfides dissolution during charge/discharge, resulting in a capacity of 940 mAh g-1 after 120 charge/discharge cycles.

  6. Aspergillus flavus Conidia-derived Carbon/Sulfur Composite as a Cathode Material for High Performance Lithium-Sulfur Battery.

    PubMed

    Xu, Maowen; Jia, Min; Mao, Cuiping; Liu, Sangui; Bao, Shujuan; Jiang, Jian; Liu, Yang; Lu, Zhisong

    2016-01-01

    A novel approach was developed to prepare porous carbon materials with an extremely high surface area of 2459.6 m(2)g(-1) by using Aspergillus flavus conidia as precursors. The porous carbon serves as a superior cathode material to anchor sulfur due to its uniform and tortuous morphology, enabling high capacity and good cycle lifetime in lithium sulfur-batteries. Under a current rate of 0.2 C, the carbon-sulfur composites with 56.7 wt% sulfur loading deliver an initial capacity of 1625 mAh g(-1), which is almost equal to the theoretical capacity of sulfur. The good performance may be ascribed to excellent electronic networks constructed by the high-surface-area carbon species. Moreover, the semi-closed architecture of derived carbons can effectively retard the polysulfides dissolution during charge/discharge, resulting in a capacity of 940 mAh g(-1) after 120 charge/discharge cycles. PMID:26732547

  7. ord1, an oxidoreductase gene responsible for conversion of O-methylsterigmatocystin to aflatoxin in Aspergillus flavus.

    PubMed Central

    Prieto, R; Woloshuk, C P

    1997-01-01

    Among the enzymatic steps in the aflatoxin biosynthetic pathway, the conversion of O-methylsterigmatocystin to aflatoxin has been proposed to be catalyzed by an oxidoreductase. Transformants of Aspergillus flavus 649WAF2 containing a 3.3-kb genomic DNA fragment and the aflatoxin biosynthesis regulatory gene aflR converted exogenously supplied O-methylsterigmatocystin to aflatoxin B1. A gene, ord1, corresponding to a transcript of about 2 kb was identified within the 3.3-kb DNA fragment. The promoter region presented a putative AFLR binding site and a TATA sequence. The nucleotide sequence of the gene revealed an open reading frame encoding a protein of 528 amino acids with a deduced molecular mass of 60.2 kDa. The gene contained six introns and seven exons. Heterologous expression of the ord1 open reading frame under the transcriptional control of the Saccharomyces cerevisiae galactose-inducible gal1 promoter results in the ability to convert O-methylsterigmatocystin to aflatoxin B1. The data indicate that ord1 is sufficient to accomplish the last step of the aflatoxin biosynthetic pathway. A search of various databases for similarity indicated that ord1 encodes a cytochrome P-450-type monooxygenase, and the gene has been assigned to a new P-450 gene family named CYP64. PMID:9143099

  8. Antifungal activity of metabolites from the marine sponges Amphimedon sp. and Monanchora arbuscula against Aspergillus flavus strains isolated from peanuts (Arachis hypogaea).

    PubMed

    Arevabini, Cynthia; Crivelenti, Yasmin D; de Abreu, Mariana H; Bitencourt, Tamires A; Santos, Mário F C; Berlinck, Roberto G S; Hajdu, Eduardo; Beleboni, Renê O; Fachin, Ana L; Marins, Mozart

    2014-01-01

    Contamination of preharvest and stored peanuts (Arachis hypogaea L.) by aflatoxigenic strains of Aspergillus flavus is an important economical and food safety problem in many tropical and subtropical areas of the world. The present investigation reports the antifungal activity of a halitoxins/amphitoxins enriched extract obtained from the sponge Amphimedon sp. (HAEEAsp), and of batzelladine L isolated from the sponge Monanchora arbuscula on Aspergillus flavus isolated from stored peanuts. A PCR system directed against the ITS region and aflatoxin biosynthetic pathway genes of A. flavus was applied for identification of aflatoxin producing strains. The HAEEAsp extract and batzelladine L showed minimal inhibitory concentration (MIC) in the range between 1.9 to 15.6 microg/mL and between 1.9 to 7.8 microg/mL, respectively. The minimal fungicide concentration (MFC) of HAEEAsp extract and batzelladine L was in the range between 3.9 to 31.3 microg/mL and 3.9 to 15.6 microg/mL, respectively. These results indicate that these marine alkaloids may be further explored for the development of potential lead compounds active against aflatoxigenic fungi. PMID:24660456

  9. Biotechnological advances for combating Aspergillus flavus and aflatoxin contamination in crops.

    PubMed

    Bhatnagar-Mathur, Pooja; Sunkara, Sowmini; Bhatnagar-Panwar, Madhurima; Waliyar, Farid; Sharma, Kiran Kumar

    2015-05-01

    Aflatoxins are toxic, carcinogenic, mutagenic, teratogenic and immunosuppressive byproducts of Aspergillus spp. that contaminate a wide range of crops such as maize, peanut, and cotton. Aflatoxin not only affects crop production but renders the produce unfit for consumption and harmful to human and livestock health, with stringent threshold limits of acceptability. In many crops, breeding for resistance is not a reliable option because of the limited availability of genotypes with durable resistance to Aspergillus. Understanding the fungal/crop/environment interactions involved in aflatoxin contamination is therefore essential in designing measures for its prevention and control. For a sustainable solution to aflatoxin contamination, research must be focused on identifying and improving knowledge of host-plant resistance factors to aflatoxin accumulation. Current advances in genetic transformation, proteomics, RNAi technology, and marker-assisted selection offer great potential in minimizing pre-harvest aflatoxin contamination in cultivated crop species. Moreover, developing effective phenotyping strategies for transgenic as well as precision breeding of resistance genes into commercial varieties is critical. While appropriate storage practices can generally minimize post-harvest aflatoxin contamination in crops, the use of biotechnology to interrupt the probability of pre-harvest infection and contamination has the potential to provide sustainable solution. PMID:25804815

  10. Aspergillus flavus VelB acts distinctly from VeA in conidiation and may coordinate with FluG to modulate sclerotial production.

    PubMed

    Chang, Perng-Kuang; Scharfenstein, Leslie L; Li, Ping; Ehrlich, Kenneth C

    2013-01-01

    The proteins VeA, VelB and LaeA of Aspergillus nidulans form a heterotrimeric complex (the velvet complex) in the dark to coordinate sexual development and production of some secondary metabolites. VeA and VelB of A. nidulans and Aspergillus fumigatus also are repressors of conidiation, but VeA of Aspergillus flavus in studied strains acts positively on conidiation. In the present study, we show via yeast-two hybrid assays that interactions among A. flavus VeA, VelB, and LaeA are conserved as in the A. nidulans velvet complex. We found that FluG, which is required for conidiophore formation in A. nidulans but whose deletion in A. flavus delays onset of conidiation, was probably an interacting partner of VelB. Deletion of velB in A. flavus CA14 severely impaired conidiation in the dark although to a lesser extent than deletion of veA. In both mutants fluG deletion resulted in further decreased conidiation even in the light. Deletion of fluG in the ΔlaeA strain, however, did not affect conidiation. All mutant types were unable to produce aflatoxin and sclerotia. Cross-complementation of the ΔvelB strain with gpdA::veA restored conidiation but not aflatoxin production although aflR, the aflatoxin pathway regulatory gene, was expressed at a normal level. Cross-complementation of the ΔveA strain with gpdA::velB failed to restore conidiation and aflatoxin production. The ΔvelB strain complemented with or a wild type transformed by gpdA::velB had elevated sclerotial production as the ΔfluG strain. Concerted interactions of A. flavus VeA and VelB with LaeA are critical for conidiation and aflatoxin biosynthesis. VelB may have a dual role and likely coordinates with FluG to modulate sclerotial production. PMID:23994319

  11. Genome-Wide Transcriptome Analysis of Cotton (Gossypium hirsutum L.) Identifies Candidate Gene Signatures in Response to Aflatoxin Producing Fungus Aspergillus flavus

    PubMed Central

    Bedre, Renesh; Rajasekaran, Kanniah; Mangu, Venkata Ramanarao; Sanchez Timm, Luis Eduardo; Bhatnagar, Deepak; Baisakh, Niranjan

    2015-01-01

    Aflatoxins are toxic and potent carcinogenic metabolites produced from the fungi Aspergillus flavus and A. parasiticus. Aflatoxins can contaminate cottonseed under conducive preharvest and postharvest conditions. United States federal regulations restrict the use of aflatoxin contaminated cottonseed at >20 ppb for animal feed. Several strategies have been proposed for controlling aflatoxin contamination, and much success has been achieved by the application of an atoxigenic strain of A. flavus in cotton, peanut and maize fields. Development of cultivars resistant to aflatoxin through overexpression of resistance associated genes and/or knocking down aflatoxin biosynthesis of A. flavus will be an effective strategy for controlling aflatoxin contamination in cotton. In this study, genome-wide transcriptome profiling was performed to identify differentially expressed genes in response to infection with both toxigenic and atoxigenic strains of A. flavus on cotton (Gossypium hirsutum L.) pericarp and seed. The genes involved in antifungal response, oxidative burst, transcription factors, defense signaling pathways and stress response were highly differentially expressed in pericarp and seed tissues in response to A. flavus infection. The cell-wall modifying genes and genes involved in the production of antimicrobial substances were more active in pericarp as compared to seed. The genes involved in auxin and cytokinin signaling were also induced. Most of the genes involved in defense response in cotton were highly induced in pericarp than in seed. The global gene expression analysis in response to fungal invasion in cotton will serve as a source for identifying biomarkers for breeding, potential candidate genes for transgenic manipulation, and will help in understanding complex plant-fungal interaction for future downstream research. PMID:26366857

  12. Genome-Wide Transcriptome Analysis of Cotton (Gossypium hirsutum L.) Identifies Candidate Gene Signatures in Response to Aflatoxin Producing Fungus Aspergillus flavus.

    PubMed

    Bedre, Renesh; Rajasekaran, Kanniah; Mangu, Venkata Ramanarao; Sanchez Timm, Luis Eduardo; Bhatnagar, Deepak; Baisakh, Niranjan

    2015-01-01

    Aflatoxins are toxic and potent carcinogenic metabolites produced from the fungi Aspergillus flavus and A. parasiticus. Aflatoxins can contaminate cottonseed under conducive preharvest and postharvest conditions. United States federal regulations restrict the use of aflatoxin contaminated cottonseed at >20 ppb for animal feed. Several strategies have been proposed for controlling aflatoxin contamination, and much success has been achieved by the application of an atoxigenic strain of A. flavus in cotton, peanut and maize fields. Development of cultivars resistant to aflatoxin through overexpression of resistance associated genes and/or knocking down aflatoxin biosynthesis of A. flavus will be an effective strategy for controlling aflatoxin contamination in cotton. In this study, genome-wide transcriptome profiling was performed to identify differentially expressed genes in response to infection with both toxigenic and atoxigenic strains of A. flavus on cotton (Gossypium hirsutum L.) pericarp and seed. The genes involved in antifungal response, oxidative burst, transcription factors, defense signaling pathways and stress response were highly differentially expressed in pericarp and seed tissues in response to A. flavus infection. The cell-wall modifying genes and genes involved in the production of antimicrobial substances were more active in pericarp as compared to seed. The genes involved in auxin and cytokinin signaling were also induced. Most of the genes involved in defense response in cotton were highly induced in pericarp than in seed. The global gene expression analysis in response to fungal invasion in cotton will serve as a source for identifying biomarkers for breeding, potential candidate genes for transgenic manipulation, and will help in understanding complex plant-fungal interaction for future downstream research. PMID:26366857

  13. Improvement of the antifungal activity of Litsea cubeba vapor by using a helium-neon (He-Ne) laser against Aspergillus flavus on brown rice snack bars.

    PubMed

    Suhem, Kitiya; Matan, Narumol; Matan, Nirundorn; Danworaphong, Sorasak; Aewsiri, Tanong

    2015-12-23

    The aim of this study was to improve the antifungal activity of the volatile Litsea cubeba essential oil and its main components (citral and limonene) on brown rice snack bars by applying He-Ne laser treatment. Different volumes (50-200 μL) of L. cubeba, citral or limonene were absorbed into a filter paper and placed inside an oven (18 L). Ten brown rice snack bars (2 cm wide × 4 cm long × 0.5 cm deep) were put in an oven and heated at 180 °C for 20 min. The shelf-life of the treated snack bars at 30 °C was assessed and sensory testing was carried out to investigate their consumer acceptability. A count of total phenolic content (TPC) and Fourier transform infrared spectroscopy (FTIR) on the properties of essential oil, citral, and limonene before and after the laser treatment was studied for possible modes of action. It was found that the laser treatment improved the antifungal activity of the examined volatile L. cubeba and citral with Aspergillus flavus inhibition by 80% in comparison with those of the control not treated with the laser. L. cubeba vapor at 100 μL with the laser treatment was found to completely inhibit the growth of natural molds on the snack bars for at least 25 days; however, without essential oil vapor and laser treatment, naturally contaminating mold was observed in 3 days. Results from the sensory tests showed that the panelists were unable to detect flavor and aroma differences between essential oil treatment and the control. Laser treatment caused an increase in TPC of citral oil whereas the TPC in limonene showed a decrease after the laser treatment. These situations could result from the changing peak of the aliphatic hydrocarbons that was revealed by the FTIR spectra. PMID:26433461

  14. Transcriptome Analysis of Aspergillus flavus Reveals veA-Dependent Regulation of Secondary Metabolite Gene Clusters, Including the Novel Aflavarin Cluster

    PubMed Central

    Cary, J. W.; Han, Z.; Yin, Y.; Lohmar, J. M.; Shantappa, S.; Harris-Coward, P. Y.; Mack, B.; Ehrlich, K. C.; Wei, Q.; Arroyo-Manzanares, N.; Uka, V.; Vanhaecke, L.; Bhatnagar, D.; Yu, J.; Nierman, W. C.; Johns, M. A.; Sorensen, D.; Shen, H.; De Saeger, S.; Diana Di Mavungu, J.

    2015-01-01

    The global regulatory veA gene governs development and secondary metabolism in numerous fungal species, including Aspergillus flavus. This is especially relevant since A. flavus infects crops of agricultural importance worldwide, contaminating them with potent mycotoxins. The most well-known are aflatoxins, which are cytotoxic and carcinogenic polyketide compounds. The production of aflatoxins and the expression of genes implicated in the production of these mycotoxins are veA dependent. The genes responsible for the synthesis of aflatoxins are clustered, a signature common for genes involved in fungal secondary metabolism. Studies of the A. flavus genome revealed many gene clusters possibly connected to the synthesis of secondary metabolites. Many of these metabolites are still unknown, or the association between a known metabolite and a particular gene cluster has not yet been established. In the present transcriptome study, we show that veA is necessary for the expression of a large number of genes. Twenty-eight out of the predicted 56 secondary metabolite gene clusters include at least one gene that is differentially expressed depending on presence or absence of veA. One of the clusters under the influence of veA is cluster 39. The absence of veA results in a downregulation of the five genes found within this cluster. Interestingly, our results indicate that the cluster is expressed mainly in sclerotia. Chemical analysis of sclerotial extracts revealed that cluster 39 is responsible for the production of aflavarin. PMID:26209694

  15. Evaluation of mRNA Expression Levels of cyp51A and mdr1, Candidate Genes for Voriconazole Resistance in Aspergillus flavus

    PubMed Central

    Fattahi, Azam; Zaini, Farideh; Kordbacheh, Parivash; Rezaie, Sasan; Safara, Mahin; Fateh, Roohollah; Farahyar, Shirin; Kanani, Ali; Heidari, Mansour

    2015-01-01

    Background: Voriconazole Resistance (VRC-R) in Aspergillus flavus isolates impacts the management of aspergillosis, since azoles are the first choice for prophylaxis and therapy. However, to the best of our knowledge, the mechanisms underlying voriconazole resistance are poorly understood. Objectives: The present study was designed to evaluate mRNA expression levels of cyp51A and mdr1 genes in voriconazole resistant A. flavus by a Real-Time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) technique. Materials and Methods: Five A. flavus isolates with resistance to VRC were examined by a RT-PCR approach. Results: Four out of five isolates revealed cyp51A and mdr1 mRNA overexpression. Interestingly, the isolate, which was negative for cyp51A and mdr1 mRNA expression showed a high voriconazole Minimum Inhibitory Concentration (MIC). Furthermore, a computational-based analysis predicted that voriconazole resistance could be mediated through cooperation with a network protein interaction. Conclusions: Our experimental and in silico findings may provide new insight in the complex molecular pathways of drug resistance and also could assist design an efficient therapeutic strategy for aspergillosis treatment. PMID:26865941

  16. Regulation of aflatoxin biosynthesis and branched-chain amino acids metabolism in Aspergillus flavus by 2-phenylethanol reveal biocontrol mechanism of Pichia anomala

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pichia anomala WRL-076 is a biocontrol yeast which has been shown to inhibit growth and aflatoxin production of A. flavus. Using the SPME-GC/MS analysis we identified that the volatile, 2-phenylethanol (2-PE) produced by this yeast and demonstrated that the compound inhibited aflatoxin production. W...

  17. Use of pyrosequencing to quantify incidence of a specific Aspergillus flavus strain within complex fungal communities associated with commercial cotton crops.

    PubMed

    Das, Modan K; Ehrlich, Kenneth C; Cotty, Peter J

    2008-03-01

    Atoxigenic strains of Aspergillus flavus have been used as aflatoxin management tools on over 50,000 hectares of commercial crops since 2000. To assess treatment efficacy, atoxigenic strain incidence is routinely monitored by vegetative compatibility analyses (VCA) that require culturing, generation of auxotrophs, and complementation with tester mutants. Two pyrosequencing assays (PA) that require no culturing were developed for monitoring incidences of atoxigenic strains on ginned cottonseed. The assays, which quantify frequencies of characteristic single nucleotide polymorphisms (SNPs) in the aflR and pksA genes, were validated against standard VCA on cottonseed collected from commercial gins in South Texas, Arizona, and Southern California where the atoxigenic strain AF36 is used to manage aflatoxin contamination. Cottonseed washings were subjected to both VCA and PA. PA was performed directly on DNA isolated from particulates pelleted from the wash water by centrifugation. Addition of CaCl(2) and diatomaceous earth prior to pelleting increased the amount of DNA isolated. Accuracy and reproducibility of the PA were contrasted with those for the VCA that has been used for over a decade. Correlation coefficients between VCA and PA indicated good correspondence between the results from the two assays (r = 0.91 for aflR assay and r = 0.80 for pksA assay). PAs were highly variable for samples with low incidences of A. flavus due to variability in the initial polymerase chain reaction step. This held for both DNA isolated from cottonseed washes and for mixtures of purified DNA. For samples yielding low quantities of A. flavus DNA, averaging of results from 4 to 5 replicates was required to achieve acceptable correlations with VCA. Pyrosequencing has the potential to become a powerful tool for monitoring atoxigenic strains within complex A. flavus communities without limitations imposed by traditional culturing methods. PMID:18944078

  18. Aspergillus: introduction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Species in the genus Aspergillus possess versatile metabolic activities that impact our daily life both positively and negatively. Aspergillus flavus and Aspergillus oryzae are closely related fungi. While the former is able to produce carcinogenic aflatoxins and is an etiological agent of aspergill...

  19. The potential of Origanum vulgare L. (Lamiaceae) essential oil in inhibiting the growth of some food-related Aspergillus species

    PubMed Central

    Carmo, Egberto Santos; de Oliveira Lima, Edeltrudes; de Souza, Evandro Leite

    2008-01-01

    Origanum vulgare L. (Lamiaceae) has been currently known for their interesting antimicrobial activity being regarded as alternative antimicrobial for use is food conservation systems. This study aimed to evaluate the effectiveness of O. vulgare essential oil in inhibiting the growth of some food-related Aspergillus species (A. flavus, A. parasiticus, A. terreus, A. ochraceus, A. fumigatus and A. niger). The essential oil revealed a strong anti-Aspergillus property providing an inhibition of all assayed mould strains. MIC values were between 80 and 20 μL/mL being found a MIC50 of 40 μL/mL. The essential oil at concentration of 80 and 40 μL/mL provided a fungicidal effect on A. flavus, A. fumigatus and A. niger noted by a total inhibition of the radial mycelial growth along 14 days of interaction. In addition, the essential oil was able to inhibit the mould spores germination when assayed at concentrations of 80 and 40 μL/mL. Our results showed the interesting anti-Aspergillus activity of O. vulgare essential oil supporting their possible use as anti-mould compound in food conservation. PMID:24031231

  20. Use of UHPLC high-resolution Orbitrap mass spectrometry to investigate the genes involved in the production of secondary metabolites in Aspergillus flavus.

    PubMed

    Arroyo-Manzanares, Natalia; Di Mavungu, José Diana; Uka, Valdet; Malysheva, Svetlana V; Cary, Jeffrey W; Ehrlich, Kenneth C; Vanhaecke, Lynn; Bhatnagar, Deepak; De Saeger, Sarah

    2015-01-01

    The fungus Aspergillus flavus is known for its ability to produce the toxic and carcinogenic aflatoxins in food and feed. While aflatoxins are of most concern, A. flavus is predicted to be capable of producing many more metabolites based on a study of its complete genome sequence. Some of these metabolites could be of great importance in food and feed safety. Here we describe an analytical methodology based on Orbitrap HRMS technology that allows the untargeted determination of fungal metabolites, in support of the study of the function of genes involved in secondary metabolism in fungi. The applied strategy implies the detection and identification of differentially expressed metabolites in extracts of wild-type and mutant fungal strains, using Orbitrap high-resolution mass spectrometry (HRMS) accurate mass data. The suitability of this approach was demonstrated by the confirmation of previously characterised genes involved in the aflatoxin biosynthetic pathway, namely a polyketide synthase (pksA), an oxidoreductase (ordA) and a methyltransferase (omtA) gene. Subsequently, the proposed methodology was applied for the detection and identification of metabolites produced by a yet uncharacterised gene cluster in A. favus, cluster 23. Comparative Orbitrap HRMS analysis of extracts of A. flavus wild-type strain and an over-expression mutant for the transcription factor of gene cluster 23 (lepE) demonstrated that this gene cluster is responsible for the production a set of 2-pyridone derivatives, the leporins. Besides the known derivatives leporin B and leporin B precursor that could be identified by automatic de-replication of the accurate mass data, five other compounds belonging to this class of fungal secondary metabolites were detected and identified for the first time, combining MS and multiple-stage MS data. PMID:26278397

  1. Identification and Quantification of a Toxigenic and Non-Toxigenic Aspergillus flavus Strain in Contaminated Maize Using Quantitative Real-Time PCR

    PubMed Central

    Mylroie, J. Erik; Ozkan, Seval; Shivaji, Renuka; Windham, Gary L.; Alpe, Michael N.; Williams, W. Paul

    2016-01-01

    Aflatoxins, which are produced by Aspergillus flavus, are toxic to humans, livestock, and pets. The value of maize (Zea mays) grain is markedly reduced when contaminated with aflatoxin. Plant resistance and biological control using non-toxin producing strains are considered effective strategies for reducing aflatoxin accumulation in maize grain. Distinguishing between the toxin and non-toxin producing strains is important in determining the effectiveness of bio-control strategies and understanding inter-strain interactions. Using polymorphisms found in the fungal rRNA intergenic spacer region (IGS) between a toxigenic strain of A. flavus (NRRL 3357) and the non-toxigenic strain used in the biological control agent Afla-Guard® (NRRL 21882), we developed a set of primers that allows for the identification and quantification of the two strains using quantitative PCR. This primer set has been used to screen maize grain that was inoculated with the two strains individually and co-inoculated with both strains, and it has been shown to be effective in both the identification and quantification of both strains. Screening of co-inoculated ears from multiple resistant and susceptible genotypic crosses revealed no significant differences in fungal biomass accumulation of either strain in the field tests from 2010 and 2011 when compared across the means of all genotypes. Only one genotype/year combination showed significant differences in strain accumulation. Aflatoxin accumulation analysis showed that, as expected, genotypes inoculated with the toxigenic strain accumulated more aflatoxin than when co-inoculated with both strains or inoculated with only the non-toxigenic strain. Furthermore, accumulation of toxigenic fungal mass was significantly correlated with aflatoxin accumulation while non-toxigenic fungal accumulation was not. This primer set will allow researchers to better determine how the two fungal strains compete on the maize ear and investigate the interaction

  2. An industry perspective on the use of “atoxigenic” strains of Aspergillus flavus as biological control agents and the significance of cyclopiazonic acid

    PubMed Central

    King, Eileen D; (Bobby) Bassi, Albeit B; Ross, David C; Druebbisch, Bernd

    2011-01-01

    Several nonaflatoxigenic strains of Aspergillus flavus have been registered in the United States to reduce aflatoxin accumulation in maize and other crops, but there may be unintended negative consequences if these strains produce cyclopiazonic acid (CPA). AF36, a nonaflatoxigenic, CPA-producing strain has been shown to produce CPA in treated maize and peanuts. Alternative strains, including Afla-Guard® brand biocontrol agent and K49, do not produce CPA and can reduce both aflatoxin and CPA in treated crops. Chronic toxicity of CPA has not been studied, and recent animal studies show significant harmful effects from short-term exposure to CPA at low doses. Grower and industry confidence in this approach must be preserved through transparency. PMID:22844262

  3. A Public Platform for the Verification of the Phenotypic Effect of Candidate Genes for Resistance to Aflatoxin Accumulation and Aspergillus flavus Infection in Maize

    PubMed Central

    Warburton, Marilyn L.; Williams, William Paul; Hawkins, Leigh; Bridges, Susan; Gresham, Cathy; Harper, Jonathan; Ozkan, Seval; Mylroie, J. Erik; Shan, Xueyan

    2011-01-01

    A public candidate gene testing pipeline for resistance to aflatoxin accumulation or Aspergillus flavus infection in maize is presented here. The pipeline consists of steps for identifying, testing, and verifying the association of selected maize gene sequences with resistance under field conditions. Resources include a database of genetic and protein sequences associated with the reduction in aflatoxin contamination from previous studies; eight diverse inbred maize lines for polymorphism identification within any maize gene sequence; four Quantitative Trait Loci (QTL) mapping populations and one association mapping panel, all phenotyped for aflatoxin accumulation resistance and associated phenotypes; and capacity for Insertion/Deletion (InDel) and SNP genotyping in the population(s) for mapping. To date, ten genes have been identified as possible candidate genes and put through the candidate gene testing pipeline, and results are presented here to demonstrate the utility of the pipeline. PMID:22069738

  4. The Potential Inhibitory Effect of Cuminum Cyminum, Ziziphora Clinopodioides and Nigella Sativa Essential Oils on the Growth of Aspergillus Fumigatus and Aspergillus

    PubMed Central

    Khosravi, A.R.; Minooeianhaghighi, M.H.; Shokri, H.; Emami, S.A.; S.M., Alavi; Asili, J.

    2011-01-01

    The goals of this study were to evaluate the effectiveness of Cuminum cyminum, Ziziphora clinopodioides and Nigella sativa essential oils to inhibit the growth of Aspergillus fumigatus and A. flavus and to evoke ultrastructural changes. The fungi were cultured into RPMI 1640 media in the presence of oils at concentrations of 8, 6, 5, 4, 3, 2, 1.5, 1.25, 1, 0.75 and 0.5 mg/ml in broth microdilution and 2, 1.5, 1 and 0.5 mg/ml in broth macrodilution methods with shaking for 48 h at 28oC. Conidial and mycelial samples exposed to 0.25, 0.5, 1, 1.5 and 2 mg essential oils/ml for 5 days in 2% yeast extract granulated plus 15% Saccharose media were processed for transmission electron microscopy (TEM). Based on broth dilution methods, C. cyminum and to a lesser extent Z. clinopodioides oils exhibited the strongest activity against A. fumigatus and A. flavus with MIC90 ranging from 0.25 to 1.5 mg/ml, while the oil from N. sativa exhibited relatively moderate activity against two above fungi with MIC90 ranging from 1.5 to 2 mg/ml. The main changes observed by TEM were in the cell wall, plasma membrane and membranous organelles; in particular, in the nuclei and mitochondria. These modifications in fungal structure were associated with the interference of the essential oils with the enzymes responsible for cell wall synthesis, which disturbed normal growth. Moreover, the essential oils caused high vacuolation of the cytoplasm, detachment of fibrillar layer of cell wall, plasma membrane disruption and disorganization of the nuclear and mitochondrial structures. Aspergillus fumigatus and A. flavus growth inhibition induced by these oils were found to be well-correlated with subsequent morphological changes of the fungi exposed to different fungistatic concentrations of the oils. Our results show the anti-Aspergillus activities of C. cyminum, Z. clinopodioides and N. sativa essential oils, which strengthens the potential use of these substances as anti-mould in the future. PMID

  5. Effect of essential oils on Aspergillus spore germination, growth and mycotoxin production: a potential source of botanical food preservative

    PubMed Central

    Gemeda, Negero; Woldeamanuel, Yimtubezinash; Asrat, Daniel; Debella, Asfaw

    2014-01-01

    Objective To investigate effect of essential oils on Aspergillus spore germination, growth and mycotoxin production. Method In vitro antifungal and antiaflatoxigenic activity of essential oils was carried out using poisoned food techniques, spore germination assay, agar dilution assay, and aflatoxin arresting assay on toxigenic strains of Aspergillus species. Results Cymbopogon martinii, Foeniculum vulgare and Trachyspermum ammi (T. ammi) essential oils were tested against toxicogenic isolates of Aspergillus species. T. ammi oil showed highest antifungal activity. Absolute mycelial inhibition was recorded at 1 µl/mL by essential oils of T. ammi. The oil also showed, complete inhibition of spore germination at a concentration of 2 µl/mL. In addition, T. ammi oil showed significant antiaflatoxigenic potency by totally inhibiting aflatoxin production from Aspergillus niger and Aspergillus flavus at 0.5 and 0.75 µl/mL, respectively. Cymbopogon martinii, Foeniculum vulgare and T. ammi oils as antifungal were found superior over synthetic preservative. Moreover, a concentration of 5 336.297 µl/kg body weight was recorded for LC50 on mice indicating the low mammalian toxicity and strengthening its traditional reputations. Conclusions In conclusion, the essential oils from T. ammi can be a potential source of safe natural food preservative for food commodities contamination by storage fungi. PMID:25183114

  6. In vitro experimental environments lacking or containing soil disparately affect competition experiments of Aspergillus flavus and co-occurring fungi in maize grains.

    PubMed

    Falade, Titilayo D O; Syed Mohdhamdan, Sharifah H; Sultanbawa, Yasmina; Fletcher, Mary T; Harvey, Jagger J W; Chaliha, Mridusmita; Fox, Glen P

    2016-07-01

    In vitro experimental environments are used to study interactions between microorganisms, and to predict dynamics in natural ecosystems. This study highlights that experimental in vitro environments should be selected to match closely the natural environment of interest during in vitro studies to strengthen extrapolations about aflatoxin production by Aspergillus and competing organisms. Fungal competition and aflatoxin accumulation were studied in soil, cotton wool or tube (water-only) environments, for Aspergillus flavus competition with Penicillium purpurogenum, Fusarium oxysporum or Sarocladium zeae within maize grains. Inoculated grains were incubated in each environment at two temperature regimes (25 and 30°C). Competition experiments showed interaction between the main effects of aflatoxin accumulation and the environment at 25°C, but not so at 30°C. However, competition experiments showed fungal populations were always interacting with their environments. Fungal survival differed after the 72-h incubation in different experimental environments. Whereas all fungi incubated within the soil environment survived, in the cotton wool environment none of the competitors of A. flavus survived at 30°C. With aflatoxin accumulation, F. oxysporum was the only fungus able to interdict aflatoxin production at both temperatures. This occurred only in the soil environment and fumonisins accumulated instead. Smallholder farmers in developing countries face serious mycotoxin contamination of their grains, and soil is a natural reservoir for the associated fungal propagules, and a drying and storage surface for grains on these farms. Studying fungal dynamics in the soil environment and other environments in vitro can provide insights into aflatoxin accumulation post-harvest. PMID:27264786

  7. Genetic Analysis of the Aspergillus flavus Vegetative Compatibility Group to Which a Biological Control Agent That Limits Aflatoxin Contamination in U.S. Crops Belongs.

    PubMed

    Grubisha, Lisa C; Cotty, Peter J

    2015-09-01

    Some filamentous fungi in Aspergillus section Flavi produce carcinogenic secondary compounds called aflatoxins. Aflatoxin contamination is routinely managed in commercial agriculture with strains of Aspergillus flavus that do not produce aflatoxins. These non-aflatoxin-producing strains competitively exclude aflatoxin producers and reshape fungal communities so that strains with the aflatoxin-producing phenotype are less frequent. This study evaluated the genetic variation within naturally occurring atoxigenic A. flavus strains from the endemic vegetative compatibility group (VCG) YV36. AF36 is a strain of VCG YV36 and was the first fungus used in agriculture for aflatoxin management. Genetic analyses based on mating-type loci, 21 microsatellite loci, and a single nucleotide polymorphism (SNP) in the aflC gene were applied to a set of 237 YV36 isolates collected from 1990 through 2005 from desert legumes and untreated fields and from fields previously treated with AF36 across the southern United States. One haplotype dominated across time and space. No recombination with strains belonging to VCGs other than YV36 was detected. All YV36 isolates carried the SNP in aflC that prevents aflatoxin biosynthesis and the mat1-2 idiomorph at the mating-type locus. These results suggest that VCG YV36 has a clonal population structure maintained across both time and space. These results demonstrate the genetic stability of atoxigenic strains belonging to a broadly distributed endemic VCG in both untreated populations and populations where the short-term frequency of VCG YV36 has increased due to applications of a strain used to competitively exclude aflatoxin producers. This work supports the hypothesis that strains of this VCG are not involved in routine genetic exchange with aflatoxin-producing strains. PMID:26092465

  8. Genetic Analysis of the Aspergillus flavus Vegetative Compatibility Group to Which a Biological Control Agent That Limits Aflatoxin Contamination in U.S. Crops Belongs

    PubMed Central

    Cotty, Peter J.

    2015-01-01

    Some filamentous fungi in Aspergillus section Flavi produce carcinogenic secondary compounds called aflatoxins. Aflatoxin contamination is routinely managed in commercial agriculture with strains of Aspergillus flavus that do not produce aflatoxins. These non-aflatoxin-producing strains competitively exclude aflatoxin producers and reshape fungal communities so that strains with the aflatoxin-producing phenotype are less frequent. This study evaluated the genetic variation within naturally occurring atoxigenic A. flavus strains from the endemic vegetative compatibility group (VCG) YV36. AF36 is a strain of VCG YV36 and was the first fungus used in agriculture for aflatoxin management. Genetic analyses based on mating-type loci, 21 microsatellite loci, and a single nucleotide polymorphism (SNP) in the aflC gene were applied to a set of 237 YV36 isolates collected from 1990 through 2005 from desert legumes and untreated fields and from fields previously treated with AF36 across the southern United States. One haplotype dominated across time and space. No recombination with strains belonging to VCGs other than YV36 was detected. All YV36 isolates carried the SNP in aflC that prevents aflatoxin biosynthesis and the mat1-2 idiomorph at the mating-type locus. These results suggest that VCG YV36 has a clonal population structure maintained across both time and space. These results demonstrate the genetic stability of atoxigenic strains belonging to a broadly distributed endemic VCG in both untreated populations and populations where the short-term frequency of VCG YV36 has increased due to applications of a strain used to competitively exclude aflatoxin producers. This work supports the hypothesis that strains of this VCG are not involved in routine genetic exchange with aflatoxin-producing strains. PMID:26092465

  9. Vitality Stains and Real Time PCR Studies to Delineate the Interactions of Pichia anomala and Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were to probe the effect of the yeast, P. anomala against A flavus by using real time RT-PCR technique and vitality fluorescent stains. Yeast and fungi were inoculated into a 250 ml-flask containing 50 ml potato dextrose broth (PDB) at yeast to fungus (Y : F) ratios of ...

  10. Application of loop-mediated isothermal amplification assays for direct identification of pure cultures of Aspergillus flavus, A. nomius, and A. caelatus and for their rapid detection in shelled Brazil nuts.

    PubMed

    Luo, Jie; Taniwaki, Marta H; Iamanaka, Beatriz T; Vogel, Rudi F; Niessen, Ludwig

    2014-02-17

    Brazil nuts have a high nutritional content and are a very important trade commodity for some Latin American countries. Aflatoxins are carcinogenic fungal secondary metabolites. In Brazil nuts they are produced predominantly by Aspergillus (A.) nomius and A. flavus. In the present study we applied and evaluated two sets of primers previously published for the specific detection of the two species using loop-mediated isothermal amplification (LAMP) technology. Moreover, a primer set specific for A. caelatus as a frequently occurring non-aflatoxigenic member of Aspergillus section Flavi in Brazil nuts was newly developed. LAMP assays were combined with a simplified DNA release method and used for rapid identification of pure cultures and rapid detection of A. nomius and A. flavus from samples of shelled Brazil nuts. An analysis of pure cultures of 68 isolates representing the major Aspergillus species occurring on Brazil nuts showed that the three LAMP assays had individual accuracies of 61.5%, 84.4%, and 93.3% for A. flavus, A. nomius, and A. caelatus, respectively when morphological identification was used as a reference. The detection limits for conidia added directly to the individual LAMP reactions were found to be 10⁵ conidia per reaction with the primer set ID9 for A. nomius and 10⁴ conidia per reaction with the primer set ID58 for A. flavus. Sensitivity was increased to 10¹ and 10² conidia per reaction for A. nomius and A. flavus, respectively, when sample preparation included a spore disruption step. The results of LAMP assays obtained during the analysis of 32 Brazil nut samples from different regions of Brazil and from different steps in the production process of the commodity were compared with results obtained from mycological analysis and aflatoxin analysis of corresponding samples. Compared with mycological analysis of the samples, the Negative Predictive Values of LAMP assays were 42.1% and 12.5% while the Positive Predictive Values were 61

  11. Environmental distribution and genetic diversity of vegetative compatibility groups determine biocontrol strategies to mitigate aflatoxin contamination of maize by Aspergillus flavus.

    PubMed

    Atehnkeng, Joseph; Donner, Matthias; Ojiambo, Peter S; Ikotun, Babatunde; Augusto, Joao; Cotty, Peter J; Bandyopadhyay, Ranajit

    2016-01-01

    Maize infected by aflatoxin-producing Aspergillus flavus may become contaminated with aflatoxins, and as a result, threaten human health, food security and farmers' income in developing countries where maize is a staple. Environmental distribution and genetic diversity of A. flavus can influence the effectiveness of atoxigenic isolates in mitigating aflatoxin contamination. However, such information has not been used to facilitate selection and deployment of atoxigenic isolates. A total of 35 isolates of A. flavus isolated from maize samples collected from three agro-ecological zones of Nigeria were used in this study. Ecophysiological characteristics, distribution and genetic diversity of the isolates were determined to identify vegetative compatibility groups (VCGs). The generated data were used to inform selection and deployment of native atoxigenic isolates to mitigate aflatoxin contamination in maize. In co-inoculation with toxigenic isolates, atoxigenic isolates reduced aflatoxin contamination in grain by > 96%. A total of 25 VCGs were inferred from the collected isolates based on complementation tests involving nitrate non-utilizing (nit(-)) mutants. To determine genetic diversity and distribution of VCGs across agro-ecological zones, 832 nit(-) mutants from 52 locations in 11 administrative districts were paired with one self-complementary nitrate auxotroph tester-pair for each VCG. Atoxigenic VCGs accounted for 81.1% of the 153 positive complementations recorded. Genetic diversity of VCGs was highest in the derived savannah agro-ecological zone (H = 2.61) compared with the southern Guinea savannah (H = 1.90) and northern Guinea savannah (H = 0.94) zones. Genetic richness (H = 2.60) and evenness (E5  = 0.96) of VCGs were high across all agro-ecological zones. Ten VCGs (40%) had members restricted to the original location of isolation, whereas 15 VCGs (60%) had members located between the original source of isolation and a distance

  12. Natural control of corn postharvest fungi Aspergillus flavus and Penicillium sp. using essential oils from plants grown in Argentina.

    PubMed

    Camiletti, Boris X; Asensio, Claudia M; Pecci, María de la Paz Giménez; Lucini, Enrique I

    2014-12-01

    The objective in this study was to evaluate the antifungal activity of essential oils from native and commercial aromatic plants grown in Argentina against corn postharvest fungi and to link the essential oil bioactivity with lipid oxidation and morphological changes in fungus cell membrane. Essential oil (EO) of oregano variety Mendocino (OMen), Cordobes (OCor), and Compacto (OCom), mint variety Inglesa (Mi), and Pehaujo (Mp), Suico (Sui); rosemary (Ro), and Aguaribay (Ag) were tested in vitro against 4 corn fungi: A. flavus (CCC116-83 and BXC01), P. oxalicum (083296), and P. minioluteum (BXC03). The minimum fungicidal concentration (MFC) and the minimum inhibitory concentration (MIC) were determined. The chemical profiles of the EOs were analyzed by GC-MS. Lipid oxidation in cell membrane of fungi was determined by hydroperoxides and related with essential oil antifungal activity. The major compounds were Thymol in OCor (18.66%), Omen (12.18%), and OCom (9.44%); menthol in Mi and Mp; verbenone in Sui; dehydroxy-isocalamendiol in Ag; and eucaliptol in Ro. OCor, Omen, and OCom showed the best antifungal activity. No antifungal activity was observed in Ag and Ro EO. The hydroperoxide value depended on the fungi (P < 0.001) and the antimicrobial agent (P < 0.001).Membrane lipids were oxidized by Sui EO in A. flavus BXC01 and A. flavus CCC116-83 (0.021 and 0.027 meqO2 /kg, respectively). The results suggest that the EOs of OCor, OMen, OCom, Mi, Mp, and Sui grown in Argentina can be used as natural alternatives to control fungi that produce mycotoxin in maize. PMID:25376651

  13. Efficacy of Some Essential Oils Against Aspergillus flavus with Special Reference to Lippia alba Oil an Inhibitor of Fungal Proliferation and Aflatoxin B1 Production in Green Gram Seeds during Storage.

    PubMed

    Pandey, Abhay K; Sonker, Nivedita; Singh, Pooja

    2016-04-01

    During mycofloral analysis of green gram (Vigna radiata (L.) R. Wilczek) seed samples taken from different grocery stores by agar and standard blotter paper methods, 5 fungal species were identified, of which Aspergillus flavus exhibited higher relative frequency (75.20% to 80.60%) and was found to produce aflatoxin B1 . On screening of 11 plant essential oils against this mycotoxigenic fungi, Lippia alba essential oil was found to be most effective and showed absolute inhibition of mycelia growth at 0.28 μL/mL. The oil of L. alba was fungistatic and fungicidal at 0.14 and 0.28 μL/mL, respectively. Oil had broad range of fungitoxicity at its MIC value and was absolutely inhibited the AFB1 production level at 2.0 μL/mL. Chemical analysis of this oil revealed geranial (36.9%) and neral (29.3%) as major components followed by myrcene (18.6%). Application of a dose of 80 μL/0.25 L air of Lippia oil in the storage system significantly inhibited the fungal proliferation and aflatoxin production without affecting the seed germination rate. By the virtue of fungicidal, antiaflatoxigenic nature and potent efficacy in storage food system, L. alba oil can be commercialized as botanical fungicide for the protection of green gram seeds during storage. PMID:26928885

  14. High incidence of Aspergillus flavus and aflatoxins in stored groundnut in Ghana and the use of a microbial assay to assess the inhibitory effects of plant extracts on aflatoxin synthesis.

    PubMed

    Awuah, R T; Kpodo, K A

    1996-01-01

    Groundnut samples from 21 selected markets in the 10 regions of Ghana yielded high levels of the aflatoxigenic fungus Aspergillus flavus on half-strength potato dextrose agar. The fungus was associated with 31.7 and 12.8%, respectively, of all damaged and undamaged kernels assayed. Only 0.24% of total kernels assayed yielded A. parasiticus. Other fungi detected from total kernels assayed were A. niger (34%), A. candidus (1.45%), A. tamarii (3.93%), A. ochraceous (5.26%), Fusarium spp. (1.7%) Penicillium spp. (5.19%), a Mucor sp. (2.3%), a Trichoderma sp. (0.2%), Rhizopus stolonifer (12%) and certain unidentifiable fungi (11.72%). Total aflatoxin levels ranging from 5.7 to 22, 168 ppb were identified with damaged kernel samples. The mycotoxin was not detected in 50% of undamaged kernel samples tested and very low levels mostly ranging from 0.1 to 12.2 ppb were associated with the undamaged samples that tested positive for aflatoxins. In a novel in vitro microbial assay to determine the effectiveness of certain plant extracts against aflatoxin synthesis, extracts from Xylopia aethiopica, Monodera myristica, Cinnamomum verum and Piper nigrum permitted fungal growth in 1.5% potato-dextrose broth while completely suppressing NOR formation. These extracts, however, could not suppress NOR formation in a yeast extract sucrose medium. PMID:8981776

  15. Characterization of species of the Aspergillus section Nigri from corn field isolates co-infected with Aspergillus flavus/parasiticus species and the potential for ochratoxin A production.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Members of the Aspergillus section Nigri, known as black-spored aspergilli, can contaminate several substrates including maize. Although some species within the group can produce plant disease symptoms such as black mold in onions and maize ear rot, the main concern with A. niger aggregate contamina...

  16. Absence of the Aflatoxin Biosynthesis Gene, norA, allows accumulation of deoxyaflatoxin B1 in Aspergillus flavus cultures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Conversion of O-methylsterigmatocystin (OMST) to aflatoxin B1 (AFB1), a highly toxic and carcinogenic fungal metabolite of some Aspergillus species, begins with its oxidation catalyzed by the cytochrome P450 monooxygenase, OrdA (AflQ). The complexity of the subsequent oxidation, hydration, ring-ope...

  17. Genome Sequences of Eight Aspergillus flavus spp. and One A. parasiticus sp., Isolated from Peanut Seeds in Georgia.

    PubMed

    Faustinelli, Paola C; Wang, Xinye Monica; Palencia, Edwin R; Arias, Renée S

    2016-01-01

    Aspergillus flavusandA. parasiticusfungi produce carcinogenic mycotoxins in peanut seeds, causing considerable impact on both human health and the economy. Here, we report nine genome sequences ofAspergillusspp., isolated from Georgia peanut seeds in 2014. The information obtained will lead to further biodiversity studies that are essential for developing control strategies. PMID:27081142

  18. Beyond aflatoxin: four distinct expression patterns and functional roles associated with Aspergillus flavus secondary metabolism gene clusters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Species of Aspergillus produce a diverse array of secondary metabolites, and recent genomic analysis has predicted that these species have the capacity to synthesize many more compounds. It has been possible to infer the presence of 55 gene clusters associated with secondary metabolism in Aspergill...

  19. Effects of Temperature, Water Activity, and Incubation Time on Production of Aflatoxins and Cyclopiazonic Acid by an Isolate of Aspergillus flavus in Surface Agar Culture

    PubMed Central

    Gqaleni, N.; Smith, J. E.; Lacey, J.; Gettinby, G.

    1997-01-01

    An experiment with a full factorial design was used to study the effects of and interactions among temperature, water activity (a(infw)), incubation period, and substrate on coproduction of aflatoxins (AF) and cyclopiazonic acid (CPA) by an isolate of Aspergillus flavus. Analysis of variance showed that there was a complex interaction among all of these factors and that this influenced the relative concentrations of the mycotoxins produced. The optimum temperatures for the production of AF and CPA were 30(deg)C and 25(deg)C, respectively. Both mycotoxins were maximally produced (0.306 to 0.330 (mu)g of AF(middot)ml of medium(sup-1), 4.040 to 6.256 (mu)g of CPA(middot)ml of medium(sup-1)) at an a(infw) of 0.996 and after 15 days of incubation. No AF were produced in either yeast extract agar or Czapek yeast autolysate agar medium at an a(infw) of 0.90 at 20 or 37(deg)C after 15 days (minimum conditions), while 0.077 to 0.439 (mu)g of CPA(middot)ml of medium(sup-1) was produced under the same conditions. Yeast extract agar favored maximum AF production, and Czapek yeast autolysate agar favored maximum CPA production. PMID:16535539

  20. Effect of cinnamomum zeylanicum blume essential oil on the growth and morphogenesis of some potentially pathogenic Aspergillus species

    PubMed Central

    Carmo, Egberto Santos; de Oliveira Lima, Edeltrudes; de Souza, Evandro Leite; de Sousa, Frederico Barbosa

    2008-01-01

    Cinnamomum zeylanicum Blume is known for a wide range of medicinal properties. This study aimed to assess the interference of C. zeylanicum essential oil on the growth and morphogenesis of some potentially pathogenic Aspergillus species. The essential oil presented strong antifungal effect causing the growth inhibition of the assayed strains and development of large growth inhibition zones. MIC50 and MIC90 values were 40 and 80 μL/mL, respectively. 80, 40 and 20 μL/mL of the oil strongly inhibited the radial mycelial growth of A. niger, A. flavus and A. fumigatus along 14 days. 80 and 40 μL/mL of the oil caused a 100% inhibition of the fungal spore germination. Main morphological changes observed under light microscopy provided by the essential oil in the fungal strains were decreased conidiation, leakage of cytoplasm, loss of pigmentation and disrupted cell structure indicating fungal wall degeneration. It is concluded that C. zeylanicum essential oil could be known as potential antifungal compound, particularly, to protect against the growth of Aspergillus species. PMID:24031186

  1. Microarray-Based Mapping for the Detection of Molecular Markers in Response to Aspergillus flavus Infection in Susceptible and Resistant Maize Lines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were (1) to evaluate differential gene expression levels for resistance to A. flavus kernel infection in susceptible (Va35) and resistant (Mp313E) maize lines using Oligonucleotide and cDNA microarray analysis, (2) to evaluate differences in A. flavus accumulation betwee...

  2. Atypical Aspergillus parasiticus isolates from pistachio with aflR gene nucleotide insertion identical to Aspergillus sojae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are the most toxic and carcinogenic secondary metabolites produced primarily by the filamentous fungi Aspergillus flavus and Aspergillus parasiticus. The toxins cause devastating economic losses because of strict regulations on distribution of contaminated products. Aspergillus sojae are...

  3. VeA is associated with the response to oxidative stress in the aflatoxin producer Aspergillus flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Survival of fungal species depends on the ability of these organisms to respond to environmental stresses. Osmotic stress or high levels of reactive oxygen species (ROS) can cause stress in fungi resulting in growth inhibition. Both eukaryotic and prokaryotic cells have developed numerous mechanisms...

  4. Based on biochemical and physiological behavior, where is Aspergillus egyptiacus better placed?

    PubMed

    Zohri, A A; Ismail, M A

    1994-01-01

    Physiological and biochemical properties were tested in 45 isolates of Aspergillus egyptiacus (16 isolates), Emericella nidulans (16) and Aspergillus versicolor (13). The three fungal species exhibited common and similar features. The big similarity between A. egyptiacus and E. nidulans was greater than between A. egyptiacus and A. versicolor. It included the inability to produce base either from sodium citrate or lactic acid media, growth at 45 degrees C (thermophilicity), and production of very similar pigmentations on Aspergillus flavus and parasiticus agar. A. egyptiacus is therefore better placed in the Aspergillus nidulans-Emericella assemblage. PMID:7537240

  5. Effect of Cymbopogon martinii, Foeniculum vulgare, and Trachyspermum ammi Essential Oils on the Growth and Mycotoxins Production by Aspergillus Species

    PubMed Central

    Woldeamanuel, Yimtubezinash; Asrat, Daniel; Debella, Asfaw

    2014-01-01

    This study was performed to investigate effect of essential oils on Aspergillus spore germination, growth, and mycotoxin production. In vitro antifungal and antiaflatoxigenic activities of Cymbopogon martinii, Foeniculum vulgare, and Trachyspermum ammi essential oils were carried out on toxigenic strains of Aspergillus species. Plant materials were hydrodistilled for 4-5 h in Clevenger apparatus. 0.25 μL/mL, 0.5 μL/mL, 1 μL/mL, 2 μL/mL, and 4 μL/mL concentrations of each essential oil were prepared in 0.1% Tween 80 (V/V). T. ammi oil showed highest antifungal activity. Absolute mycelial inhibition was recorded at 1 μL/mL by essential oils of T. ammi. The oil also showed complete inhibition of spore germination at a concentration of 2 μL/mL. In addition, T. ammi oil showed significant antiaflatoxigenic potency by totally inhibiting toxin production from A. niger and A. flavus at 0.5 and 0.75 μL/mL, respectively. C. martinii, F. vulgare, and T. ammi oils as antifungals were found superior over synthetic preservative. Moreover, a concentration of 5336.297 μL/kg body weight was recorded for LC50 on mice indicating the low mammalian toxicity. In conclusion, the essential oils from T. ammi can be a potential source of safe natural food preservative for food commodities contamination by Aspergillus species. PMID:26904653

  6. Detection of Aspergillus flavus and A. fumigatus in Bronchoalveolar Lavage Specimens of Hematopoietic Stem Cell Transplants and Hematological Malignancies Patients by Real-Time Polymerase Chain Reaction, Nested PCR and Mycological Assays

    PubMed Central

    Zarrinfar, Hossein; Mirhendi, Hossein; Fata, Abdolmajid; Khodadadi, Hossein; Kordbacheh, Parivash

    2015-01-01

    Background: Pulmonary aspergillosis (PA) is one of the most serious complications in immunocompromised patients, in particular among hematopoietic stem cell transplants (HSCT) and patients with hematological malignancies. Objectives: The current study aimed to evaluate the incidence of PA and utility of molecular methods in HSCT and patients with hematological malignancies, four methods including direct examination, culture, nested polymerase chain reaction (PCR) and real-time PCR were performed on bronchoalveolar lavage (BAL) specimens in Tehran, Iran. Patients and Methods: During 16 months, 46 BAL specimens were obtained from individuals with allogeneic HSCT (n = 18) and patients with hematological malignancies (n = 28). Direct wet mounts with 20% potassium hydroxide (KOH) and culture on mycological media were performed. The molecular detection of Aspergillus fumigatus and A. flavus was done by amplifying the conserved sequences of internal transcribed spacer 1 (ITS1) ribosomal DNA by nested-PCR and the β-tubulin gene by TaqMan real-time PCR. Results: Seven (15.2%) out of 46 specimens were positive in direct examination and showed branched septate hyphae; 11 (23.9%) had positive culture including eight (72.7%) A. flavus and three (27.3%) A. fumigatus; 22 (47.8%) had positive nested-PCR and eight (17.4%) had positive real-time PCR. The incidence of invasive pulmonary aspergillosis (IPA) in these patients included proven IPA in 1 (2.2%), probable IPA in 10 (21.7%), possible IPA in 19 (41.3%) and not IPA in 16 cases (34.8%). Conclusions: The incidence of IPA in allogeneic HSCT and patients with hematological malignancies was relatively high and A. flavus was the most common cause of PA. As molecular methods had higher sensitivity, it may be useful as screening methods in HSCT and patients with hematological malignancies, or to determine when empirical antifungal therapy can be withheld. PMID:25763133

  7. Effect of Carum copticum essential oil on growth and aflatoxin formation by Aspergillus strains.

    PubMed

    Kazemi, M

    2015-01-01

    The objectives of this study were to determine the antiaflatoxin B1 activity in vitro of the essential oil (EO) extracted from the seeds of Carum copticum and to evaluate its antifungal activity in vivo as a potential food preservative. The C. copticum EO exhibited noticeable inhibition on dry mycelium and synthesis of aflatoxin B1 (AFB1) by Aspergillus flavus, completely inhibiting AFB1 production at 4 μL/mL. C. copticum EOs showed the lowest percentages of decayed cherry tomatoes for all fungi compared with the control at 100 μL/mL with values of 5.01 ± 67% for A. flavus and 5.98 ± 54% for Aspergillus niger. The results indicated that the percentage of infected fruits is significantly (p < 0.01) reduced by the EO at 16°C for 30 days. In this case, the oil at 100 μL/mL concentration showed the highest inhibition of fungal infection with a value of 80.45% compared with the control. Thus, the EO of dill could be used to control food spoilage and as a potential source of food preservative. PMID:25342249

  8. Localization of growth and secretion of proteins in Aspergillus niger.

    PubMed

    Wösten, H A; Moukha, S M; Sietsma, J H; Wessels, J G

    1991-08-01

    Hyphal growth and secretion of proteins in Aspergillus niger were studied using a new method of culturing the fungus between perforated membranes which allows visualization of both parameters. At the colony level the sites of occurrence of growth and general protein secretion were correlated. In 4-d-old colonies both growth and secretion were localized at the periphery of the colony, whereas in a 5-d-old colony growth and secretion also occurred in a more central zone of the colony where conidiophore differentiation was observed. However, in both cases glucoamylase secretion was mainly detected at the periphery of the colonies. At the hyphal level immunogold labelling showed glucoamylase secretion at the tips of leading hyphae only. Microautoradiography after labelling with N-acetylglucosamine showed that these hyphae were probably all growing. Glucoamylase secretion could not be demonstrated immediately after a temperature shock which stopped growth. These results indicate that glucoamylase secretion is located at the tips of growing hyphae only. PMID:1955876

  9. Expression of Aspergillus hemoglobin domain activities in Aspergillus oryzae grown on solid substrates improves growth rate and enzyme production.

    PubMed

    te Biesebeke, Rob; Boussier, Amandine; van Biezen, Nick; Braaksma, Machtelt; van den Hondel, Cees A M J J; de Vos, Willem M; Punt, Peter J

    2006-01-01

    DNA fragments coding for hemoglobin domains (HBD) were isolated from Aspergillus oryzae and Aspergillus niger. The HBD activities were expressed in A. oryzae by introduction of HBD gene fragments under the control of the promoter of the constitutively expressed gpdA gene. In the transformants, oxygen uptake was significantly higher, and during growth on solid substrates the developed biomass was at least 1.3 times higher than that of the untransformed wild-type strain. Growth rate of the HBD-activity-producing strains was also significantly higher compared to the wild type. During growth on solid cereal substrates, the amylase and protease activities in the extracts of the HBD-activity-producing strains were 30-150% higher and glucoamylase activities were at least 9 times higher compared to the wild-type strain. These results suggest that the Aspergillus HBD-encoding gene can be used in a self-cloning strategy to improve biomass yield and protein production of Aspergillus species. PMID:16927259

  10. Modelling the effect of temperature and water activity in the growth boundaries of Aspergillus ochraceus and Aspergillus parasiticus.

    PubMed

    Garcia, Daiana; Ramos, Antonio J; Sanchis, Vicente; Marín, Sonia

    2011-05-01

    The aim of this work was to model the growth of Aspergillus parasiticus and Aspergillus ochraceus, both mycotoxin producers, near to the growth/no growth boundaries and validate those models in sterile maize grain, peanuts and coffee beans. Malt extract agar was adjusted to six different water activities: 0.93, 0.91, 0.89, 0.87, 0.85 and 0.80. Plates were incubated at 10, 15, 20, 25, 30, 37 and 42 °C. For each of the 42 conditions, 10 Petri dishes were inoculated. Both kinetic and probability models were applied to colony growth data. The results of the present study indicate that the developed probability modelling approach could be satisfactorily employed to quantify the combined effect of temperature and water activity on the growth responses of A. ochraceus and A. parasiticus. However, validation of kinetic results led to poor goodness of prediction. In this study, the validation samples were placed near to the expected boundaries of the models in order to test them under the worst situation. Probability of growth prediction under extreme growth conditions was somewhat compromised, but it can be considered acceptable. PMID:21356445

  11. Aspergillus flavus VelB acts distinctly from VeA in conidiation and may coordinate with FluG to modulate sclerotial production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asexual and sexual differentiation in Aspergillus nidulans involve complex control by a number of factors and is light-dependent. The velvet protein, VeA, in A. nidulans is a negative regulator of conidiation and a positive regulator of sexual development. It forms a complex with VelB and LaeA to co...

  12. On the Permeation by Dioxygen of Urate Oxidase from Aspergillus flavus in Complex with Xanthine Anion: Dioxygen Pathways and a Portrait of the Enzyme Cavities from Molecular Dynamics Simulations in Water Solution.

    PubMed

    Pietra, Francesco

    2016-06-01

    This work describes molecular dynamics (MD) simulations in aqueous media for the complex of the homotetrameric urate oxidase (UOX) from Aspergillus flavus with xanthine anion (5) in the presence of dioxygen (O2 ). After 196.6 ns of trajectory from unrestrained MD, a O2 molecule was observed leaving the bulk solvent to penetrate the enzyme between two subunits, A/C. From here, the same O2 molecule was observed migrating, across subunit C, to the hydrophobic cavity that shares residue V227 with the active site. The latter was finally attained, after 378.3 ns of trajectory, with O2 at a bonding distance from 5. The reverse same O2 pathway, from 5 to the bulk solvent, was observed as preferred pathway under random acceleration MD (RAMD), where an external, randomly oriented force was acting on O2 . Both MD and RAMD simulations revealed several cavities populated by O2 during its migration from the bulk solvent to the active site or backwards. Paying attention to the last hydrophobic cavity that apparently serves as O2 reservoir for the active site, it was noticed that its volume undergoes ample fluctuations during the MD simulation, as expected from the thermal motion of a flexible protein, independently from the particular subunit and no matter whether the cavity is filled or not by O2 . PMID:27151738

  13. Effect of different light wavelengths on the growth and ochratoxin A production in Aspergillus carbonarius and Aspergillus westerdijkiae.

    PubMed

    Cheong, Khai Khuang; Strub, Caroline; Montet, Didier; Durand, Noël; Alter, Pascaline; Meile, Jean-Christophe; Schorr Galindo, Sabine; Fontana, Angélique

    2016-05-01

    The effects of light at different wavelengths and photoperiod on growth and ochratoxin A production of Aspergillus carbonarius and Aspergillus westerdijkiae were studied: far-red (740 nm), red (625 nm), blue (445 nm), and UV-A (366 nm). Fungal growth was not significantly affected by photoperiod or light wavelength; the only exception was A. westerdijkiae which showed reduced growth under UV-A light (366 nm). Short-wavelength blue light (445 nm) and UV-A light caused a reduction in ochratoxin A production of both fungal species. However, long-wavelength red light (625 nm) and far-red light (740 nm) reduced ochratoxin A production only in A. westerdijkiae but not in A. carbonarius. It is believed that this difference in reactivity to light is due to differences in the melanin content of the two fungal species: A. carbonarius is a black fungus with higher melanin content than A. westerdijkiae, a yellow fungus. Other possible explanations for the reduction of ochratoxin A production by light were also discussed. PMID:27109370

  14. Reactive oxygen species and their possible role in defense signaling in maize-Aspergilus flavus interations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The colonization of maize by Aspergillus flavus and the contamination of kernel tissues with aflatoxins has been the subject of intensive research for >40 years. Several proteins have been identified as components of resistance against A. flavus. Many function in abiotic stress tolerance by preventi...

  15. Candida parapsilosis as a Potent Biocontrol Agent against Growth and Aflatoxin Production by Aspergillus Species

    PubMed Central

    Niknejad, F; Zaini, F; Faramarzi, MA; Amini, M; Kordbacheh, P; Mahmoudi, M; Safara, M

    2012-01-01

    Background: Aflatoxin contamination of food and feed stuff is a serious health problem and significant economic concerns. In the present study, the inhibitory effect of Candida parapsilosis IP1698 on mycelial growth and aflatoxin production in aflatoxigenic strains of Aspergillus species was investigated. Methods: Mycelial growth inhibitions of nine strains of aflatoxigenic and non-aflatoxigenic Aspergillus species in the presence of C. parapsilosis investigated by pour plate technique at different pH, temperature and time of incubation. Reduction of aflatoxin was evaluated in co-cultured fungi in yeast extract sucrose broth after seven days of incubation using HPLC method. The data were analyzed by SPSS 11.5. Results: The presence of the C. parapsilosis at different pH did not affect significantly the growth rate of Aspergillus isolates. On the other hand, temperature and time of incubation showed to be significantly effective when compared to controls without C. parapsilosis (P≤0.05). In aflatoxigenic strains, minimum percentage of reductions in total aflatoxin and B1, B2, G1, G2 fractions were 92.98, 92.54, 77.48, 54.54 and 72.22 and maximum percentage of reductions were 99.59, not detectable, 94.42, and not detectable in both G1 and G2, respectively. Conclusion: C. parapsilosis might employ as a good biocontrol agent against growth and aflatoxin production by aflatoxigenic Aspergillus species PMID:23308351

  16. 2-hydroxyisocaproic acid is fungicidal for Candida and Aspergillus species.

    PubMed

    Sakko, M; Moore, C; Novak-Frazer, L; Rautemaa, V; Sorsa, T; Hietala, P; Järvinen, A; Bowyer, P; Tjäderhane, L; Rautemaa, R

    2014-04-01

    The amino acid derivative 2-hydroxyisocaproic acid (HICA) is a nutritional additive used to increase muscle mass. Low levels can be detected in human plasma as a result of leucine metabolism. It has broad antibacterial activity but its efficacy against pathogenic fungi is not known. The aim was to test the efficacy of HICA against Candida and Aspergillus species. Efficacy of HICA against 19 clinical and reference isolates representing five Candida and three Aspergillus species with variable azole antifungal sensitivity profiles was tested using a microdilution method. The concentrations were 18, 36 and 72 mg ml(-1) . Growth was determined spectrophotometrically for Candida isolates and by visual inspection for Aspergillus isolates, viability was tested by culture and impact on morphology by microscopy. HICA of 72 mg ml(-1) was fungicidal against all Candida and Aspergillus fumigatus and Aspergillus terreus isolates. Lower concentrations were fungistatic. Aspergillus flavus was not inhibited by HICA. HICA inhibited hyphal formation in susceptible Candida albicans and A. fumigatus isolates and affected cell wall integrity. In conclusion, HICA has broad antifungal activity against Candida and Aspergillus at concentrations relevant for topical therapy. As a fungicidal agent with broad-spectrum bactericidal activity, it may be useful in the topical treatment of multispecies superficial infections. PMID:24125484

  17. Human polymorphonuclear leukocytes inhibit Aspergillus fumigatus conidial growth by lactoferrin-mediated iron depletion.

    PubMed

    Zarember, Kol A; Sugui, Janyce A; Chang, Yun C; Kwon-Chung, Kyung J; Gallin, John I

    2007-05-15

    Aspergillus fumigatus, a common mold, rarely infects humans, except during prolonged neutropenia or in cases of chronic granulomatous disease (CGD), a primary immunodeficiency caused by mutations in the NADPH oxidase that normally produces fungicidal reactive oxygen species. Filamentous hyphae of Aspergillus are killed by normal, but not CGD polymorphonuclear leukocytes (PMN); however, the few studies on PMN-mediated host defenses against infectious conidia (spores) of this organism have yielded conflicting results, some showing that PMN do not inhibit conidial growth, with others showing that they do, most likely using reactive oxygen species. Given that CGD patients are exposed daily to hundreds of viable A. fumigatus conidia, yet considerable numbers of them survive years without infection, we reasoned that PMN use ROS-independent mechanisms to combat Aspergillus. We show that human PMN from both normal controls and CGD patients are equipotent at arresting the growth of Aspergillus conidia in vitro, indicating the presence of a reactive oxygen species-independent factor(s). Cell-free supernatants of degranulated normal and CGD neutrophils both suppressed fungal growth and were found to be rich in lactoferrin, an abundant PMN secondary granule protein. Purified iron-poor lactoferrin at concentrations occurring in PMN supernatants (and reported in human mucosal secretions in vivo) decreased fungal growth, whereas saturation of lactoferrin or PMN supernatants with iron, or testing in the presence of excess iron in the form of ferritin, completely abolished activity against conidia. These results demonstrate that PMN lactoferrin sequestration of iron is important for host defense against Aspergillus. PMID:17475866

  18. Inhibitory effects of spices on growth and toxin production of toxigenic fungi.

    PubMed Central

    Hitokoto, H; Morozumi, S; Wauke, T; Sakai, S; Kurata, H

    1980-01-01

    The inhibitory effects of 29 commercial powdered spices on the growth and toxin production of three species of toxigenic Aspergillus were observed by introducing these materials into culture media for mycotoxin production. Of the 29 samples tested, cloves, star anise seeds, and allspice completely inhibited the fungal growth, whereas most of the others inhibited only the toxin production. Eugenol extracted from cloves and thymol from thyme caused complete inhibition of the growth of both Aspergillus flavus and Aspergillus versicolor at 0.4 mg/ml or less. At a concentration of 2 mg/ml, anethol extracted from star anise seeds inhibited the growth of all the strains. PMID:6769391

  19. Biological control of AFB1-producing Aspergillus section Flavi strains isolated from brewer's grains, alternative feed intended for swine production in Argentina.

    PubMed

    Asurmendi, Paula; García, María J; Ruíz, Francisco; Dalcero, Ana; Pascual, Liliana; Barberis, Lucila

    2016-07-01

    The aim of the present study was to investigate the inhibitory activity of lactic acid bacteria (LAB) isolated from brewer's grains on Aspergillus section Flavi growth and aflatoxin B1 production. The Aspergillus strains tested were inhibited by all the LAB strains assayed. The isolates Lactobacillus brevis B20, P. pentosaceus B86, Lactococcus lactis subsp. lactis B87, L. brevis B131, and Lactobacillus sp. B144 completely suppressed the fungal growth and reduced aflatoxin B1 production. In conclusion, LAB isolated from brewer's grains show a high inhibitory activity on fungal growth and aflatoxin biosynthesis by Aspergillus flavus and Aspergillus parasiticus. Further studies must be conducted to evaluate the success of in vitro assays under food environment conditions and to elucidate the antifungal mechanism of these strains. PMID:27070819

  20. Biocontrol of Aspergillus flavus by Pichia anomala

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are extremely potent natural carcinogens and a major food safety concern because of potential contamination of food commodities. Threshold levels set by the U. S. Food and Drug Administration for aflatoxins in foods for domestic consumption are less than 20 parts/ billion (ppb). However, ...

  1. Biodiversity of Aspergillus flavus in the USA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This report serves to document research conducted under a CRADA with Circle One Global, Inc. Additional details of research can be found in the report for the parent project 6604-42000-007-00D, Biochemical, Physical, Microbiological Management for Prevention of Mycotoxins in Peanuts. The efficacy of...

  2. Effect of ultraviolet radiation A and B on growth and mycotoxin production by Aspergillus carbonarius and Aspergillus parasiticus in grape and pistachio media.

    PubMed

    García-Cela, Esther; Marin, Sonia; Sanchis, Vicente; Crespo-Sempere, Ana; Ramos, Antonio J

    2015-01-01

    The effects of two exposure times per day (6 and 16 h) of UV-A or UV-B radiation, combined with dark and dark plus light incubation periods during 7-21 d on fungal growth and mycotoxins production of Aspergillus species were studied. Aspergillus carbonarius and Aspergillus parasiticus were inoculated on grape and pistachio media under diurnal and nocturnal temperatures choosing light photoperiod according to harvest conditions of these crops in Spain. Ultraviolet irradiation had a significant effect on A. carbonarius and A. parasiticus colony size (diameter, biomass dry weight, and colony density) and mycotoxin accumulation, although intraspecies differences were observed. Inhibition of A. carbonarius fungal growth decreased when exposure time was reduced from 16 h to 6 h, but this was not always true for ochratoxin A (OTA) production. OTA reduction was higher under UV-A than UV-B radiation and the reduction increased along time conversely to the aflatoxins (AFs). Aflatoxin B1 (AFB1) was the main toxin produced by A. parasiticus except in the UV-B light irradiated colonies which showed a higher percentage of AFG than AFB. Morphological changes were observed in colonies grown under UV-B light. PMID:25601150

  3. Suppression of Growth Rate of Colony-Associated Fungi by High Fructose Corn Syrup Feeding Supplement, Formic Acid, and Oxalic Acid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Select colony-associated fungi (bee isolates). Absidia sp., Ascosphaera apis, Aspergillus flavus, Fusarium sp., Penicillium glabrum, Mucor sp., showed a 40% reduction in radial growth rate with formic acid, a 28% reduction with oxalic acid, and a 15% reduction with fructose and high fructose corn sy...

  4. Isolate-Dependent Growth, Virulence, and Cell Wall Composition in the Human Pathogen Aspergillus fumigatus

    PubMed Central

    Amarsaikhan, Nansalmaa; O’Dea, Evan M.; Tsoggerel, Angar; Owegi, Henry; Gillenwater, Jordan; Templeton, Steven P.

    2014-01-01

    The ubiquitous fungal pathogen Aspergillus fumigatus is a mediator of allergic sensitization and invasive disease in susceptible individuals. The significant genetic and phenotypic variability between and among clinical and environmental isolates are important considerations in host-pathogen studies of A. fumigatus-mediated disease. We observed decreased radial growth, rate of germination, and ability to establish colony growth in a single environmental isolate of A. fumigatus, Af5517, when compared to other clinical and environmental isolates. Af5517 also exhibited increased hyphal diameter and cell wall β-glucan and chitin content, with chitin most significantly increased. Morbidity, mortality, lung fungal burden, and tissue pathology were decreased in neutropenic Af5517-infected mice when compared to the clinical isolate Af293. Our results support previous findings that suggest a correlation between in vitro growth rates and in vivo virulence, and we propose that changes in cell wall composition may contribute to this phenotype. PMID:24945802

  5. Genetic diversity of Aspergillus species isolated from onychomycosis and Aspergillus hongkongensis sp. nov., with implications to antifungal susceptibility testing.

    PubMed

    Tsang, Chi-Ching; Hui, Teresa W S; Lee, Kim-Chung; Chen, Jonathan H K; Ngan, Antonio H Y; Tam, Emily W T; Chan, Jasper F W; Wu, Andrea L; Cheung, Mei; Tse, Brian P H; Wu, Alan K L; Lai, Christopher K C; Tsang, Dominic N C; Que, Tak-Lun; Lam, Ching-Wan; Yuen, Kwok-Yung; Lau, Susanna K P; Woo, Patrick C Y

    2016-02-01

    Thirteen Aspergillus isolates recovered from nails of 13 patients (fingernails, n=2; toenails, n=11) with onychomycosis were characterized. Twelve strains were identified by multilocus sequencing as Aspergillus spp. (Aspergillus sydowii [n=4], Aspergillus welwitschiae [n=3], Aspergillus terreus [n=2], Aspergillus flavus [n=1], Aspergillus tubingensis [n=1], and Aspergillus unguis [n=1]). Isolates of A. terreus, A. flavus, and A. unguis were also identifiable by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The 13th isolate (HKU49(T)) possessed unique morphological characteristics different from other Aspergillus spp. Molecular characterization also unambiguously showed that HKU49(T) was distinct from other Aspergillus spp. We propose the novel species Aspergillus hongkongensis to describe this previously unknown fungus. Antifungal susceptibility testing showed most Aspergillus isolates had low MICs against itraconazole and voriconazole, but all Aspergillus isolates had high MICs against fluconazole. A diverse spectrum of Aspergillus species is associated with onychomycosis. Itraconazole and voriconazole are probably better drug options for Aspergillus onychomycosis. PMID:26658315

  6. Effects of Clitoria ternatea leaf extract on growth and morphogenesis of Aspergillus niger.

    PubMed

    Kamilla, L; Mansor, S M; Ramanathan, S; Sasidharan, S

    2009-08-01

    Clitoria ternatea is known for its antimicrobial activity but the antifungal effects of leaf extract on growth and morphogenesis of Aspergillus niger have not been observed. The extract showed a favorable antifungal activity against A. niger with a minimum inhibition concentration 0.8 mg/mL and minimum fungicidal concentration 1.6 mg/mL, respectively. The leaf extract exhibited considerable antifungal activity against filamentous fungi in a dose-dependent manner with 0.4 mg/mL IC50 value on hyphal growth of A. niger. The main changes observed under scanning electron microscopy after C. ternatea extract treatment were loss of cytoplasm in fungal hyphae and the hyphal wall and its diameter became markedly thinner, distorted, and resulted in cell wall disruption. In addition, conidiophore alterations were also observed when A. niger was treated with C. ternatea leaf extract. PMID:19575837

  7. Distinct Roles of Myosins in Aspergillus fumigatus Hyphal Growth and Pathogenesis.

    PubMed

    Renshaw, Hilary; Vargas-Muñiz, José M; Richards, Amber D; Asfaw, Yohannes G; Juvvadi, Praveen R; Steinbach, William J

    2016-05-01

    Myosins are a family of actin-based motor proteins found in many organisms and are categorized into classes based on their structures. Class II and V myosins are known to be important for critical cellular processes, including cytokinesis, endocytosis, exocytosis, and organelle trafficking, in the model fungi Saccharomyces cerevisiae and Aspergillus nidulans However, the roles of myosins in the growth and virulence of the pathogen Aspergillus fumigatus are unknown. We constructed single- and double-deletion strains of the class II and class V myosins in A. fumigatus and found that while the class II myosin (myoB) is dispensable for growth, the class V myosin (myoE) is required for proper hyphal extension; deletion of myoE resulted in hyperbranching and loss of hyphal polarity. Both myoB and myoE are necessary for proper septation, conidiation, and conidial germination, but only myoB is required for conidial viability. Infection with the ΔmyoE strain in the invertebrate Galleria mellonella model and also in a persistently immunosuppressed murine model of invasive aspergillosis resulted in hypovirulence, while analysis of bronchoalveolar lavage fluid revealed that tumor necrosis factor alpha (TNF-α) release and cellular infiltration were similar compared to those of the wild-type strain. The ΔmyoE strain showed fungal growth in the murine lung, while the ΔmyoB strain exhibited little fungal burden, most likely due to the reduced conidial viability. These results show, for the first time, the important role these cytoskeletal components play in the growth of and disease caused by a known pathogen, prompting future studies to understand their regulation and potential targeting for novel antifungal therapies. PMID:26953327

  8. Clonality and sex impact aflatoxigenicity in Aspergillus populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Species in Aspergillus section Flavi commonly infect agricultural staples such as corn, peanuts, cottonseed, and tree nuts and produce an array of mycotoxins, the most potent of which are aflatoxins. Aspergillus flavus is the dominant aflatoxin-producing species in the majority of crops. Populatio...

  9. Volatile Compounds Emitted by Pseudomonas aeruginosa Stimulate Growth of the Fungal Pathogen Aspergillus fumigatus

    PubMed Central

    Briard, Benoit; Heddergott, Christoph

    2016-01-01

    ABSTRACT Chronic lung infections with opportunistic bacterial and fungal pathogens are a major cause of morbidity and mortality especially in patients with cystic fibrosis. Pseudomonas aeruginosa is the most frequently colonizing bacterium in these patients, and it is often found in association with the filamentous fungus Aspergillus fumigatus. P. aeruginosa is known to inhibit the growth of A. fumigatus in situations of direct contact, suggesting the existence of interspecies communication that may influence disease outcome. Our study shows that the lung pathogens P. aeruginosa and A. fumigatus can interact at a distance via volatile-mediated communication and expands our understanding of interspecific signaling in microbial communities. PMID:26980832

  10. Energetics of growth of Aspergillus tamarii in a biological real-time reaction calorimeter.

    PubMed

    Dhandapani, Balaji; Mahadevan, Surianarayanan; Mandal, Asit Baran

    2012-03-01

    Fungal cultivation in a biological real-time reaction calorimeter (BioRTCal) is arduous due to the heterogeneous nature of the system and difficulty in optimizing the process variables. The aim of this investigation is to monitor the growth of fungi Aspergillus tamarii MTCC 5152 in a calorimeter. Experiments carried out with a spore concentration of 10(5) spores/mL indicate that the growth based on biomass and heat generation profiles was comparable to those obtained hitherto. Heat yield due to biomass growth, substrate uptake, and oxygen uptake rate was estimated from calorimetric experiments. The results would be useful in fermenter design and scale-up. Heat of combustion of fungal biomass was determined experimentally and compared to the four models reported so far. The substrate concentration had significant effects on pellet formation with variation in pellet porosity and apparent density. Metabolic heat generation is an online process variable portraying the instantaneous activity of monitoring fungal growth and BioRTCal is employed to measure the exothermic heat in a noninvasive way. PMID:22113563

  11. Unfolded protein response is required for Aspergillus oryzae growth under conditions inducing secretory hydrolytic enzyme production.

    PubMed

    Tanaka, Mizuki; Shintani, Takahiro; Gomi, Katsuya

    2015-12-01

    Unfolded protein response (UPR) is an intracellular signaling pathway for adaptation to endoplasmic reticulum (ER) stress. In yeast UPR, Ire1 cleaves the unconventional intron of HAC1 mRNA, and the functional Hac1 protein translated from the spliced HAC1 mRNA induces the expression of ER chaperone genes and ER-associated degradation genes for the refolding or degradation of unfolded proteins. In this study, we constructed an ireA (IRE1 ortholog) conditionally expressing strain of Aspergillus oryzae, a filamentous fungus producing a large amount of amylolytic enzymes, and examined the contribution of UPR to ER stress adaptation under physiological conditions. Repression of ireA completely blocked A. oryzae growth under conditions inducing the production of hydrolytic enzymes, such as amylases and proteases. This growth defect was restored by the introduction of unconventional intronless hacA (hacA-i). Furthermore, UPR was observed to be induced by amylolytic gene expression, and the disruption of the transcriptional activator for amylolytic genes resulted in partial growth restoration of the ireA-repressing strain. In addition, a homokaryotic ireA disruption mutant was successfully generated using the strain harboring hacA-i as a parental host. These results indicated that UPR is required for A. oryzae growth to alleviate ER stress induced by excessive production of hydrolytic enzymes. PMID:26496881

  12. Systems Analysis Unfolds the Relationship between the Phosphoketolase Pathway and Growth in Aspergillus nidulans

    PubMed Central

    Regueira, Torsten B.; Hofmann, Gerald; Nielsen, Jens; Olsson, Lisbeth

    2008-01-01

    Background Aspergillus nidulans is an important model organism for studies on fundamental eukaryotic cell biology and on industrial processes due to its close relation to A. niger and A. oryzae. Here we identified the gene coding for a novel metabolic pathway in A. nidulans, namely the phosphoketolase pathway, and investigated the role of an increased phosphoketolase activity. Methodology/Principal Findings Over-expression of the phosphoketolase gene (phk) improved the specific growth rate on xylose, glycerol and ethanol. Transcriptome analysis showed that a total of 1,222 genes were significantly affected by over-expression of the phk, while more than half of the affected genes were carbon source specific. During growth on glucose medium, the transcriptome analysis showed that the response to phk over-expression is targeted to neutralize the effect of the over-expression by regulating the acetate metabolism and initiate a growth dampening response. Conclusions/Significance Metabolic flux analysis using 13C-labelled glucose, showed that over-expression of phosphoketolase added flexibility to the central metabolism. Our findings further suggests that A. nidulans is not optimized for growth on xylose, glycerol or ethanol as the sole carbon sources. PMID:19052639

  13. Dynamics of Actin Cables in Polarized Growth of the Filamentous Fungus Aspergillus nidulans

    PubMed Central

    Bergs, Anna; Ishitsuka, Yuji; Evangelinos, Minoas; Nienhaus, G. U.; Takeshita, Norio

    2016-01-01

    Highly polarized growth of filamentous fungi requires a continuous supply of proteins and lipids to the hyphal tip. This transport is managed by vesicle trafficking via the actin and microtubule cytoskeletons and their associated motor proteins. Particularly, actin cables originating from the hyphal tip are essential for hyphal growth. Although, specific marker proteins have been developed to visualize actin cables in filamentous fungi, the exact organization and dynamics of actin cables has remained elusive. Here, we observed actin cables using tropomyosin (TpmA) and Lifeact fused to fluorescent proteins in living Aspergillus nidulans hyphae and studied the dynamics and regulation. GFP tagged TpmA visualized dynamic actin cables formed from the hyphal tip with cycles of elongation and shrinkage. The elongation and shrinkage rates of actin cables were similar and approximately 0.6 μm/s. Comparison of actin markers revealed that high concentrations of Lifeact reduced actin dynamics. Simultaneous visualization of actin cables and microtubules suggests temporally and spatially coordinated polymerization and depolymerization between the two cytoskeletons. Our results provide new insights into the molecular mechanism of ordered polarized growth regulated by actin cables and microtubules. PMID:27242709

  14. Dynamics of Actin Cables in Polarized Growth of the Filamentous Fungus Aspergillus nidulans.

    PubMed

    Bergs, Anna; Ishitsuka, Yuji; Evangelinos, Minoas; Nienhaus, G U; Takeshita, Norio

    2016-01-01

    Highly polarized growth of filamentous fungi requires a continuous supply of proteins and lipids to the hyphal tip. This transport is managed by vesicle trafficking via the actin and microtubule cytoskeletons and their associated motor proteins. Particularly, actin cables originating from the hyphal tip are essential for hyphal growth. Although, specific marker proteins have been developed to visualize actin cables in filamentous fungi, the exact organization and dynamics of actin cables has remained elusive. Here, we observed actin cables using tropomyosin (TpmA) and Lifeact fused to fluorescent proteins in living Aspergillus nidulans hyphae and studied the dynamics and regulation. GFP tagged TpmA visualized dynamic actin cables formed from the hyphal tip with cycles of elongation and shrinkage. The elongation and shrinkage rates of actin cables were similar and approximately 0.6 μm/s. Comparison of actin markers revealed that high concentrations of Lifeact reduced actin dynamics. Simultaneous visualization of actin cables and microtubules suggests temporally and spatially coordinated polymerization and depolymerization between the two cytoskeletons. Our results provide new insights into the molecular mechanism of ordered polarized growth regulated by actin cables and microtubules. PMID:27242709

  15. Inhibitory Effects of Silver Nanoparticles on Growth and Aflatoxin B1 Production by Aspergillus Parasiticus

    PubMed Central

    Mousavi, Seyyed Amin Ayatollahi; Pourtalebi, Somayyeh

    2015-01-01

    Background: Aflatoxins (AFs) are secondary hazardous fungal metabolites that are produced by strains of some Aspergillus species on food and feedstuffs. Aflatoxin B1 (AFB1) is one of the most important AF with high toxicity. Prevention of AF production and their elimination from food products is a matter of importance for many researchers in the last decades. Nanomaterials applications in medical science have been widely studied in the recent years. Most of existing researches seek the effect of nanoparticles on bacteria, fungi, and viruses. The aim of this study was to determine the effects of silver nanoparticles (AgNPs) on growth and AFB1 production of AF-producing Aspergillus parasiticus. Methods: A parasiticus was inoculated (106 conidia per ml of medium) to potato dextrose broth (PDB) medium and then AgNPs was added and incubated with shaking at 130 rpm and 28°C for 7 days. AF was assayed by high performance liquid chromatography (HPLC). Microbiological assay (MBA) on microplates contained potato dextrose broth (PDB) medium (4 days at 28°C) at different concentrations of AgNPs (60, 80, 100, 120, 140, 160, 180 and 200 μg/ml) was measured. Results: The results demonstrated that a minimum inhibition concentration (MIC) equal to 180 μg/ml was determined for AgNPs against A. parasiticus. The AgNPs effectively inhibited AFB1 production at a concentration of 90 μg/ml. Conclusion: The results obtained in this study show AgNPs at concentrations lower than the MIC drastically inhibited production of AFB1 by A. parasiticus in culture medium. The AgNPs may be useful to control AF contamination of susceptible crops in the field. PMID:26538778

  16. Aspergillus nidulans ArfB Plays a Role in Endocytosis and Polarized Growth ▿ †

    PubMed Central

    Lee, Soo Chan; Schmidtke, Sabrina N.; Dangott, Lawrence J.; Shaw, Brian D.

    2008-01-01

    Filamentous fungi undergo polarized growth throughout most of their life cycles. The Spitzenkörper is an apical organelle composed primarily of vesicles that is unique to filamentous fungi and is likely to act as a vesicle supply center for tip growth. Vesicle assembly and trafficking are therefore important for hyphal growth. ADP ribosylation factors (Arfs), a group of small GTPase proteins, play an important role in nucleating vesicle assembly. Little is known about the role of Arfs in filamentous hyphal growth. We found that Aspergillus nidulans is predicted to encode six Arf family proteins. Analysis of protein sequence alignments suggests that A. nidulans ArfB shares similarity with ARF6 of Homo sapiens and Arf3p of Saccharomyces cerevisiae. An arfB null allele (arfB disrupted by a transposon [arfB::Tn]) was characterized by extended isotropic growth of germinating conidia followed by cell lysis or multiple, random germ tube emergence, consistent with a failure to establish polarity. The mutant germ tubes and hyphae that do form initially meander abnormally off of the axis of polarity and frequently exhibit dichotomous branching at cell apices, consistent with a defect in polarity maintenance. FM4-64 staining of the arfB::Tn strain revealed that another phenotypic characteristic seen for arfB::Tn is a reduction and delay in endocytosis. ArfB is myristoylated at its N terminus. Green fluorescent protein-tagged ArfB (ArfB::GFP) localizes to the plasma membrane and endomembranes and mutation (ArfBG2A::GFP) of the N-terminal myristoylation motif disperses the protein to the cytoplasm rather than to the membranes. These results demonstrate that ArfB functions in endocytosis to play important roles in polarity establishment during isotropic growth and polarity maintenance during hyphal extension. PMID:18539885

  17. Isolation of culturable mycobiota from agricultural soils and determination of tolerance to glyphosate of nontoxigenic Aspergillus section Flavi strains.

    PubMed

    Carranza, Cecilia S; Barberis, Carla L; Chiacchiera, Stella M; Dalcero, Ana María; Magnoli, Carina E

    2016-01-01

    Glyphosate-based herbicides are extensively used in Argentina's agricultural system to control undesirable weeds. This study was conducted to evaluate the culturable mycobiota [colony forming units (CFU) g(-1) and frequency of fungal genera or species] from an agricultural field exposed to pesticides. In addition, we evaluated the tolerance of A. oryzae and nontoxigenic A. flavus strains to high concentrations (100 to 500 mM - 17,000 to 84,500 ppm) of a glyphosate commercial formulation. The analysis of the mycobiota showed that the frequency of the main fungal genera varied according to the analyzed sampling period. Aspergillus spp. or Aspergillus section Flavi strains were isolated from 20 to 100% of the soil samples. Sterilia spp. were also observed throughout the sampling (50 to 100%). Aspergillus section Flavi tolerance assays showed that all of the tested strains were able to develop at the highest glyphosate concentration tested regardless of the water availability conditions. In general, significant reductions in growth rates were observed with increasing concentrations of the herbicide. However, a complete inhibition of fungal growth was not observed with the concentrations assayed. This study contributes to the knowledge of culturable mycobiota from agricultural soils exposed to pesticides and provides evidence on the effective growth ability of A. oryzae and nontoxigenic A. flavus strains exposed to high glyphosate concentrations in vitro. PMID:26549415

  18. Secondary Metabolite Profiling, Growth Profiles and Other Tools for Species Recognition and Important Aspergillus Mycotoxins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Species in the genus Aspergillus have been classified primarily based on morphological features. Sequencing of house-hold genes has also been used in Aspergillus taxonomy and phylogeny, while extrolites and physiological features have been used less frequently. Three independent ways of classifyin...

  19. Extracellular xylanolytic and pectinolytic hydrolase production by A. flavus isolates contributes to crop invasion

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several atoxigenic Aspergillus flavus isolates, including some biocontrol agents, and one toxigenic isolate were surveyed for the ability to produce extracellular xylanolytic and pectinolytic hydrolases. All of the tested isolates displayed good production of endoxylanases when grown on a medium ut...

  20. Identification of a major xylanase from A flavus as a 14-kD protein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus K49 secreted at least two xylanase activities when grown on a medium containing larch (wood) xylan as a sole carbon source. Enzyme activity was assayed using an agar medium containing Remazol Brilliant Blue R conjugated oat spelt xylan as substrate. Crude enzyme preparations wer...

  1. Effect of naturally occurring antimicrobials and chemical preservatives on the growth of Aspergillus Parasiticus.

    PubMed

    Pillai, Prathesha; Ramaswamy, K

    2012-04-01

    Effect of water activity (aw, 0.99), pH (4.5) and their interaction on the growth inhibition of Aspergillus parasiticus was studied on potato dextrose agar (PDA) using various antimicrobial agents (citral, carvacrol, eugenol, cineole, thymol guaiacol, vanillin, anethol, potassium sorbate and sorbic acid). The results demonstrate that colony diameter (mm) exhibited a constant increase with time (zero order kinetics) for all antimicrobials evaluated. Eugenol and sorbic acid inhibited the test fungi at 300 and 600 ppm, respectively. Radial growth rate (RGR) of A. parasiticus was significantly (p < 0.05) different among different antimicrobials as well as the concentrations tested. However, this difference was not observed with higher concentration of citral, eugenol, vanillin and sorbic acid. Among the antimicrobials evaluated potassium sorbate, cineole, anethol and guaiacol were least effective. Thymol, eugenol and carvacrol were more effective in inhibiting A. parasiticus even with low concentration (150 ppm) as their mean RGR was zero even after 20 days of incubation (pH 4.5). PMID:23572846

  2. Human Neutrophils Are Primed by Chemoattractant Gradients for Blocking the Growth of Aspergillus fumigatus.

    PubMed

    Jones, Caroline N; Dimisko, Laurie; Forrest, Kevin; Judice, Kevin; Poznansky, Mark C; Markmann, James F; Vyas, Jatin M; Irimia, Daniel

    2016-02-01

    The contribution of human neutrophils to the protection against fungal infections by Aspergillus fumigatus is essential but not fully understood. Whereas healthy people can inhale spores of A. fumigatus without developing disease, neutropenic patients and those receiving immunosuppressive drugs have a higher incidence of invasive fungal infections. To study the role of neutrophils in protection against A. fumigatus infections, we developed an in vitro assay in which the interactions between human neutrophils and A. fumigatus were observed in real time, at single-cell resolution, in precisely controlled conditions. We measured the outcomes of neutrophil-fungus interactions and found that human neutrophils have a limited ability to migrate toward A. fumigatus and block the growth of A. fumigatus conidia (proportion with growth blocked, 69%). The blocking ability of human neutrophils increased to 85.1% when they were stimulated by uniform concentrations of fMLP and was enhanced further, to 99.4%, in the presence of chemoattractant gradients. Neutrophils from patients receiving immunosuppressive treatment after transplantation were less effective against the fungus than those from healthy donors, and broader heterogeneity exists between patients, compared with healthy individuals. Further studies using this microfluidic platform will help understand the relevance of innate immune deficiencies responsible for the higher risk of fungal infections in patients with immunosuppressive disease. PMID:26272935

  3. Role of growth media and chemical enhancers in secondary metabolites production from Aspergillus carbonarius (NRL-369) and their pharmaceutical potentials.

    PubMed

    Khan, Abid Ali; Bacha, Nafess; Ahmad, Bashir; Cox, R J; Bakht, Jehan

    2016-07-01

    The present study investigates the effect of different growth media and chemical enhancer on silent genes in Aspergillus carbonarius (NRL-369) for secondary metabolites production and its in vitro biological activities. Results revealed that Aspergillus carbonarius (NRL-369) grown in Czapeak yeast extract broth medium produced more metabolites compared with other media. Chemical epigenetic modifiers (suberoyl-anilide hydroxamic acid (SAHA) and 5-azacytidine (5-AZA) at concentration of 15mM were effective for the expression of silent genes resulting in increased secondary metabolites production. Secondary metabolites extracted in ethyl acetate and fractionized in n-Hexane showed variable degree of growth inhibitions of the tested microorganisms. Similarly, these samples were also active against brine shrimps and Lemna. PMID:27393435

  4. Influence of the antimicrobial compound allyl isothiocyanate against the Aspergillus parasiticus growth and its aflatoxins production in pizza crust.

    PubMed

    Quiles, Juan M; Manyes, Lara; Luciano, Fernando; Mañes, Jordi; Meca, Giuseppe

    2015-09-01

    Aflatoxins (AFs) are secondary metabolites produced by different species of Aspergillus, such as Aspergillus flavus and Aspergillus parasiticus, which possess mutagenic, teratogenic and carcinogenic activities in humans. In this study, active packaging devices containing allyl isothiocyanate (AITC) or oriental mustard flour (OMF) + water were tested to inhibit the growth of A. parasiticus and AFs production in fresh pizza crust after 30 d. The antimicrobial and anti-aflatoxin activities were compared to a control group (no antimicrobial treatment) and to a group added with commercial preservatives (sorbic acid + sodium propionate). A. parasiticus growth was only inhibited after 30 d by AITC in filter paper at 5 μL/L and 10 μL/L, AITC sachet at 5 μL/L and 10 μL/L and OMF sachet at 850 mg + 850 μL of water. However, AFs production was inhibited by all antimicrobial treatments in a dose-dependent manner. More importantly, AITC in a filter paper at 10 μL/L, AITC sachet at 10 μL/L, OMF sachet at 850 mg + 850 μL of water and sorbic acid + sodium propionate at 0.5-2.0 g/Kg completely inhibited AFs formation. The use of AITC in active packaging devices could be a natural alternative to avoid the growth of mycotoxinogenic fungi in refrigerated bakery products in substitution of common commercial preservatives. PMID:26146190

  5. The allergenicity of Aspergillus fumigatus conidia is influenced by growth temperature.

    PubMed

    Low, Swee Yang; Dannemiller, Karen; Yao, Maosheng; Yamamoto, Naomichi; Peccia, Jordan

    2011-07-01

    Common indoor and outdoor environmental fungi such as Aspergillus fumigatus produce asexual spores containing a collection of proteins that can bind IgE antibodies and trigger allergic reactions. We characterized the impact of sporulation temperature on the IgE-binding capacity (allergenicity) of A. fumigatus and explored the links between variable allergenicity and temperature-dependant expression of genes encoding these allergenic proteins. A 12-fold increase in A. fumigatus allergenicity per spore was observed when sporulation temperatures were decreased from 32°C to 17°C. Per spore protein mass and Asp f 1 allergen mass also followed this trend. Functional gene expression analysis of A. fumigatus sporulating cultures by real-time reverse-transcription PCR and gene expression microarrays revealed that a greater number of genes encoding known, major allergens are more highly expressed at lower sporulation temperatures. The results of this study indicate that environmental conditions at growth significantly influence the allergenicity of this common mould through the differential production of allergenic proteins, and highlight the importance of in vivo or in vitro allergenicity measurements for understanding environmental exposure to airborne allergenic fungi. PMID:21724168

  6. Inhibitory Effect of Essential Oils on Aspergillus ochraceus Growth and Ochratoxin A Production

    PubMed Central

    Selvaraj, Jonathan Nimal; Wang, Yan; Zhao, Yueju; Zhou, Lu; Liu, Xiao; Liu, Yang

    2014-01-01

    Ochratoxin A (OTA) is a mycotoxin which is a common contaminant in grains during storage. Aspergillus ochraceus is the most common producer of OTA. Essential oils play a crucial role as a biocontrol in the reduction of fungal contamination. Essential oils namely natural cinnamaldehyde, cinnamon oil, synthetic cinnamaldehyde, Litsea citrate oil, citral, eugenol, peppermint, eucalyptus, anise and camphor oils, were tested for their efficacy against A. ochraceus growth and OTA production by fumigation and contact assays. Natural cinnamaldehyde proved to be the most effective against A. ochraceus when compared to other oils. Complete fungal growth inhibition was obtained at 150–250 µL/L with fumigation and 250–500 µL/L with contact assays for cinnamon oil, natural and synthetic cinnamaldehyde, L. citrate oil and citral. Essential oils had an impact on the ergosterol biosynthesis and OTA production. Complete inhibition of ergosterol biosynthesis was observed at ≥100 µg/mL of natural cinnamaldehyde and at 200 µg/mL of citral, but total inhibition was not observed at 200 µg/mL of eugenol. But, citral and eugenol could inhibit the OTA production at ≥75 µg/mL and ≥150 µg/mL respectively, while natural cinnamaldehyde couldn’t fully inhibit OTA production at ≤200 µg/mL. The inhibition of OTA by natural cinnamaldehyde is mainly due to the reduction in fungal biomass. However, citral and eugenol could significant inhibit the OTA biosynthetic pathway. Also, we observed that cinnamaldehyde was converted to cinnamic alcohol by A. ochraceus, suggesting that the antimicrobial activity of cinnamaldehyde was mainly attributed to its carbonyl aldehyde group. The study concludes that natural cinnamaldehyde, citral and eugenol could be potential biocontrol agents against OTA contamination in storage grains. PMID:25255251

  7. Inhibitory effect of essential oils on Aspergillus ochraceus growth and ochratoxin A production.

    PubMed

    Hua, Huijuan; Xing, Fuguo; Selvaraj, Jonathan Nimal; Wang, Yan; Zhao, Yueju; Zhou, Lu; Liu, Xiao; Liu, Yang

    2014-01-01

    Ochratoxin A (OTA) is a mycotoxin which is a common contaminant in grains during storage. Aspergillus ochraceus is the most common producer of OTA. Essential oils play a crucial role as a biocontrol in the reduction of fungal contamination. Essential oils namely natural cinnamaldehyde, cinnamon oil, synthetic cinnamaldehyde, Litsea citrate oil, citral, eugenol, peppermint, eucalyptus, anise and camphor oils, were tested for their efficacy against A. ochraceus growth and OTA production by fumigation and contact assays. Natural cinnamaldehyde proved to be the most effective against A. ochraceus when compared to other oils. Complete fungal growth inhibition was obtained at 150-250 µL/L with fumigation and 250-500 µL/L with contact assays for cinnamon oil, natural and synthetic cinnamaldehyde, L. citrate oil and citral. Essential oils had an impact on the ergosterol biosynthesis and OTA production. Complete inhibition of ergosterol biosynthesis was observed at ≥ 100 µg/mL of natural cinnamaldehyde and at 200 µg/mL of citral, but total inhibition was not observed at 200 µg/mL of eugenol. But, citral and eugenol could inhibit the OTA production at ≥ 75 µg/mL and ≥ 150 µg/mL respectively, while natural cinnamaldehyde couldn't fully inhibit OTA production at ≤ 200 µg/mL. The inhibition of OTA by natural cinnamaldehyde is mainly due to the reduction in fungal biomass. However, citral and eugenol could significant inhibit the OTA biosynthetic pathway. Also, we observed that cinnamaldehyde was converted to cinnamic alcohol by A. ochraceus, suggesting that the antimicrobial activity of cinnamaldehyde was mainly attributed to its carbonyl aldehyde group. The study concludes that natural cinnamaldehyde, citral and eugenol could be potential biocontrol agents against OTA contamination in storage grains. PMID:25255251

  8. Primary cutaneous aspergillosis due to Aspergillus tamarii in an immunocompetent host

    PubMed Central

    Sharma, Sadhna; Yenigalla, Bindu Madhav; Naidu, Sujeet Kumar; Pidakala, Premalatha

    2013-01-01

    Primary cutaneous aspergillosis is a rare disease usually caused by Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus and Aspergillus ustus. It is usually seen in immunocompromised hosts, though some cases are also reported in immunocompetent hosts. We present a case of an immunocompetent farmer who presented with generalised nodules and plaques, mimicking erythema nodosum leprosum but turned out to be cutaneous aspergillosis caused by Aspergillus tamarii. The characteristic ascospores of Aspergillus species were found in skin lesions on fungus isolated in culture. The patient showed excellent response to antifungal therapy. PMID:23970496

  9. The potential hazards of Aspergillus sp. in foods and feeds, and the role of biological treatment: a review.

    PubMed

    Sheikh-Ali, Sheikh Imranudin; Ahmad, Akil; Mohd-Setapar, Siti-Hamidah; Zakaria, Zainul Akmal; Abdul-Talib, Norfahana; Khamis, Aidee Kamal; Hoque, Md Enamul

    2014-10-01

    The contamination of food and feed by Aspergillus has become a global issue with a significant worldwide economic impact. The growth of Aspergillus is unfavourable to the development of food and feed industries, where the problems happen mostly due to the presence of mycotoxins, which is a toxic metabolite secreted by most Aspergillus groups. Moreover, fungi can produce spores that cause diseases, such as allergies and asthma, especially to human beings. High temperature, high moisture, retarded crops, and poor food storage conditions encourage the growth of mold, as well as the development of mycotoxins. A variety of chemical, biological, and physical strategies have been developed to control the production of mycotoxins. A biological approach, using a mixed culture comprised of Saccharomyces cerevisiae and Lactobacillus rhamnosus resulted in the inhibition of the growth of fungi when inoculated into fermented food. The results reveal that the mixed culture has a higher potential (37.08%) to inhibit the growth of Aspergillus flavus (producer of Aflatoxin) compared to either single culture, L. rhamnosus NRRL B-442 and S. cerevisiae, which inhibit the growth by 63.07% and 64.24%, respectively. PMID:25269603

  10. Antifungal effect of Allium tuberosum, Cinnamomum cassia, and Pogostemon cablin essential oils and their components against population of Aspergillus species.

    PubMed

    Kocevski, Dragana; Du, Muying; Kan, Jianquan; Jing, Chengjun; Lačanin, Ines; Pavlović, Hrvoje

    2013-05-01

    Antifungal activity of Allium tuberosum (AT), Cinnamomum cassia (CC), and Pogostemon cablin (Patchouli, P) essential oils against Aspergillus flavus strains 3.2758 and 3.4408 and Aspergillus oryzae was tested at 2 water activity levels (aw : 0.95 and 0.98). Main components of tested essential oils were: allyl trisulfide 40.05% (AT), cinnamaldehyde 87.23% (CC), and patchouli alcohol 44.52% (P). The minimal inhibitory concentration of the plant essential oils against A. flavus strains 3.2758 and 3.4408 and A. oryzae was 250 ppm (A. tuberosum and C. cassia), whereas Patchouli essential oil inhibited fungi at concentration > 1500 ppm. The essential oils exhibited suppression effect on colony growth at all concentrations (100, 175, and 250 ppm for A. tuberosum; 25, 50, and 75 for C. cassia; 100, 250, and 500 for P. cablin essential oil). Results of the study represent a solution for possible application of essential oil of C. cassia in different food systems due to its strong inhibitory effect against tested Aspergillus species. In real food system (table grapes), C. cassia essential oil exhibited stronger antifungal activity compared to cinnamaldehyde. PMID:23647469

  11. [Influence of fly ash concentrations on the growth of Aspergillus niger and the bioleaching efficiency of heavy metals].

    PubMed

    Yang, Jie; Wang, Qun-Hui; Wang, Qi; Xue, Jun; Tian, Shu-Lei

    2008-03-01

    The bioleaching of municipal solid waste incinerator (MSWI) fly ash for metals extraction by Aspergillus niger was investigated. The influence of fly ash concentrations on the biomass concentration, the pH of suspension, the kinds of bio-produced organic acids and the metals extraction yield during the bioleaching process were studied and the leaching toxicities of fly ash before and after bioleaching were compared. The results showed that the decrease of pH was due to generated organic acids by Aspergillus niger during bioleaching, which resulted in the metals extraction from the fly ash. The alkaline and the heavy metals toxicities of fly ash inhibited the Aspergillus niger growth, which was shown as the "lag phase". When fly ash concentration was 20 g/L, the maximum biomass was 28.61 g/L (after bioleaching 192 h), and the minimum pH was 3.85 (after finished bioleaching). The bioleaching efficiency was the highest (i.e., 93.06% for Cd, around 70% for Mn, Pb and Zn, 22%, 33% and 47% for Fe, Cr and Cu, respectively). The TCLP results of the fly ash after bioleaching indicated that the leaching toxicities of the treated fly ash were far lower than the regulated levels of China. PMID:18649552

  12. Tissue-specific components of resistance to Aspergillus ear rot of maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and other Aspergillus spp. infect maize and produce aflatoxins. One control measure is the use of resistant maize lines. There are several reports of maize lines that are resistant to aflatoxin accumulation, but the mechanisms of resistance remain unknown. To gain a better unde...

  13. Calcineurin Orchestrates Hyphal Growth, Septation, Drug Resistance and Pathogenesis of Aspergillus fumigatus: Where Do We Go from Here?

    PubMed

    Juvvadi, Praveen R; Steinbach, William J

    2015-01-01

    Studies on fungal pathogens belonging to the ascomycota phylum are critical given the ubiquity and frequency with which these fungi cause infections in humans. Among these species, Aspergillus fumigatus causes invasive aspergillosis, a leading cause of death in immunocompromised patients. Fundamental to A. fumigatus pathogenesis is hyphal growth. However, the precise mechanisms underlying hyphal growth and virulence are poorly understood. Over the past 10 years, our research towards the identification of molecular targets responsible for hyphal growth, drug resistance and virulence led to the elucidation of calcineurin as a key signaling molecule governing these processes. In this review, we summarize our salient findings on the significance of calcineurin for hyphal growth and septation in A. fumigatus and propose future perspectives on exploiting this pathway for designing new fungal-specific therapeutics. PMID:26694470

  14. Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

    PubMed Central

    Costa, Beatriz de Oliveira; Nahas, Ely

    2012-01-01

    The effect of inoculation of Aspergillus flavus , Fusarium verticillioides , and Penicillium sp. in Dystrophic Red Latosol (DRL) and Eutroferric Red Latosol (ERL) soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides . The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides . The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times ( p <0.05) in soils with glucose and inoculated with the fungi (except F. verticillioides ), in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control. PMID:24031836

  15. Septic arthritis due to tubercular and Aspergillus co-infection.

    PubMed

    Kumar, Mukesh; Thilak, Jai; Zahoor, Adnan; Jyothi, Arun

    2016-01-01

    Aspergillus septic arthritis is a rare and serious medical and surgical problem. It occurs mainly in immunocompromised patients. Aspergillus fumigatus is the most common causative organism followed by Aspergillus flavus. The most common site affected is knee followed by shoulder, ankle, wrist, hip and sacroiliac joint. Debridement and voriconazole are primary treatment of articular aspergilosis. To the best of our knowledge, there are no reported cases of co-infection of tuberculosis (TB) and Aspergillus infecting joints. We report a case of co-infection of TB and A. flavus of hip and knee of a 60-year-old male, with type 2 diabetes mellitus. He was treated with debridement, intravenous voriconazole, and antitubercular drugs. PMID:27293296

  16. The effect of humidity after gamma-irradiation on aflatoxin B-1 production of A. Flavus in ground nutmeg and peanut

    NASA Astrophysics Data System (ADS)

    Hilmy, N.; Chosdu, R.; Matsuyama, A.

    1995-02-01

    The effect of humidity of 75 up to 97% after irradiation on radiosensitivity and aflatoxin B1 production of Aspergillus flavus isolated from Indonesian nutmeg were examined. Irradiation doses used were 0;0.5;1 and 3 kGy. Mould free ground nutmeg and peanut were used as the growth media, and about 10 8 of spores were used to contaminate each of the media. Aflatoxin productions were measured after having incubated 3 days up to 5 months under humidity of 91 and 97%. Prior to HPLC analysis, aflatoxin was cleaned-up using an immunoaffinity column. The results were: (1) A. flavus indicated no or almost no growth under RH of 85% or less. (2) Under 91-97% RH, growth of mycelium and toxin production were inhibited more or less by irradiation up to 1 kGy, although the effectiveness of irradiation varied with different RH and media during postirradiation incubation. (3) By 3 kGy or more, both mycelium growth and toxin production of the mould were found to be completely inhibited. (4) The production of aflatoxin in nutmeg began after having incubated for 25 and 45 days and in peanut for 3 and 6 days under 97 and 91% RH, respectively.

  17. New applications for known drugs: Human glycogen synthase kinase 3 inhibitors as modulators of Aspergillus fumigatus growth.

    PubMed

    Sebastián, Víctor; Manoli, Maria-Tsampika; Pérez, Daniel I; Gil, Carmen; Mellado, Emilia; Martínez, Ana; Espeso, Eduardo A; Campillo, Nuria E

    2016-06-30

    Invasive aspergillosis (IA) is one of the most severe forms of fungi infection. IA disease is mainly due to Aspergillus fumigatus, an air-borne opportunistic pathogen. Mortality rate caused by IA is still very high (50-95%), because of difficulty in early diagnostics and reduced antifungal treatment options, thus new and efficient drugs are necessary. The aim of this work is, using Aspergillus nidulans as non-pathogen model, to develop efficient drugs to treat IA. The recent discovered role of glycogen synthase kinase-3 homologue, GskA, in A. fumigatus human infection and our previous experience on human GSK-3 inhibitors focus our attention on this kinase as a target for the development of antifungal drugs. With the aim to identify effective inhibitors of colonial growth of A. fumigatus we use A. nidulans as an accurate model for in vivo and in silico studies. Several well-known human GSK-3β inhibitors were tested for inhibition of A. nidulans colony growth. Computational tools as docking studies and binding site prediction was used to explain the different biological profile of the tested inhibitors. Three of the five tested hGSK3β inhibitors are able to reduce completely the colonial growth by covalent bind to the enzyme. Therefore these compounds may be useful in different applications to eradicate IA. PMID:27131621

  18. Reduction of IgE binding and nonpromotion of Aspergillus flavus fungal growth by simultaneously silencing Ara h 2 and Ara h 6 in peanut.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The most potent peanut allergens, Ara h 2 and 6, were silenced in transgenic plants by RNA interference. Three independent transgenic lines were recovered after microprojectile bombardment, of which two contained single, integrated copies of the transgene. The third line contained multiple copies ...

  19. Biodiversity of Aspergillus species in some important agricultural products

    PubMed Central

    Perrone, G.; Susca, A.; Cozzi, G.; Ehrlich, K.; Varga, J.; Frisvad, J.C.; Meijer, M.; Noonim, P.; Mahakarnchanakul, W.; Samson, R.A.

    2007-01-01

    The genus Aspergillus is one of the most important filamentous fungal genera. Aspergillus species are used in the fermentation industry, but they are also responsible of various plant and food secondary rot, with the consequence of possible accumulation of mycotoxins. The aflatoxin producing A. flavus and A. parasiticus, and ochratoxinogenic A. niger, A. ochraceus and A. carbonarius species are frequently encountered in agricultural products. Studies on the biodiversity of toxigenic Aspergillus species is useful to clarify molecular, ecological and biochemical characteristics of the different species in relation to their different adaptation to environmental and geographical conditions, and to their potential toxigenicity. Here we analyzed the biodiversity of ochratoxin producing species occurring on two important crops: grapes and coffee, and the genetic diversity of A. flavus populations occurring in agricultural fields. Altogether nine different black Aspergillus species can be found on grapes which are often difficult to identify with classical methods. The polyphasic approach used in our studies led to the identification of three new species occurring on grapes: A. brasiliensis, A. ibericus, and A. uvarum. Similar studies on the Aspergillus species occurring on coffee beans have evidenced in the last five years that A. carbonarius is an important source of ochratoxin A in coffee. Four new species within the black aspergilli were also identified in coffee beans: A. sclerotioniger, A. lacticoffeatus, A. sclerotiicarbonarius, and A. aculeatinus. The genetic diversity within A. flavus populations has been widely studied in relation to their potential aflatoxigenicity and morphological variants L- and S-strains. Within A. flavus and other Aspergillus species capable of aflatoxin production, considerable diversity is found. We summarise the main recent achievements in the diversity of the aflatoxin gene cluster in A. flavus populations, A. parasiticus and the non

  20. Application of essential oils in maize grain: impact on Aspergillus section Flavi growth parameters and aflatoxin accumulation.

    PubMed

    Bluma, Romina V; Etcheverry, Miriam G

    2008-04-01

    The antifungal activity of Pimpinella anisum L. (anise), Pëumus boldus Mol (boldus), Hedeoma multiflora Benth (mountain thyme), Syzygium aromaticum L. (clove), and Lippia turbinate var. integrifolia (griseb) (poleo) essential oils (EOs) against Aspergillus section Flavi was evaluated in sterile maize grain under different water activity (a(w)) condition (0.982, 0.955, and 0.90). The effect of EOs added to maize grains on growth rate, lag phase, and aflatoxin B(1) (AFB(1)) accumulation of Aspergillus section Flavi were evaluated at different water activity conditions. The five EOs analyzed have been shown to influence lag phase and growth rate. Their efficacy depended mainly on the essential oil concentrations and substrate water activity conditions. All EOs showed significant impact on AFB(1) accumulation. This effect was closely dependent on the water activity, concentration, and incubation periods. Important reduction of AFB(1) accumulation was observed in the majority of EO treatments at 11 days of incubation. Boldus, poleo, and mountain thyme EO completely inhibited AFB(1) at 2000 and 3000 microg g(-1). Inhibition of AFB(1) accumulation was also observed when aflatoxigenic isolates grew with different concentration of EOs during 35 days. PMID:18206775

  1. Effects of Aspergillus niger fermented rapeseed meal on nutrient digestibility, growth performance and serum parameters in growing pigs.

    PubMed

    Shi, Changyou; He, Jun; Wang, Jianping; Yu, Jie; Yu, Bing; Mao, Xiangbing; Zheng, Ping; Huang, Zhiqing; Chen, Daiwen

    2016-04-01

    The aim of the present study was to investigate the influences of Aspergillus niger fermented rapeseed meal (FRSM) on growth performance and nutrient digestibility of growing pigs. A total of 72 growing pigs (body weight = 40.8 ± 2.1 kg) were used in feeding trials, lasting for up to 42 days, and were randomly allotted to one of three diets, including a corn-soybean meal control diet as well as two experimental diets containing 10% unfermented rapeseed meal (RSM) or 10% FRSM. The results showed that average daily gain and feed conversion ratio of pigs fed FRSM were superior (P < 0.05) to that of pigs fed unfermented RSM and did not differ from the control. Pigs fed control diet had higher (P < 0.05) total tract apparent digestibility for dry matter, protein, calcium and phosphorus than pigs fed unfermented RSM diet and did not differ from the FRSM diet. Pigs fed FRSM had lower levels (P < 0.05) of serum aspartate transaminase compared to unfermented RSM. In conclusion, solid state fermentation using Aspergillus niger may improve the growth performance and nutrient digestibility of RSM for pigs and FRSM is a promising alternative protein for pig production. PMID:26434567

  2. In Vitro Interactions between Target of Rapamycin Kinase Inhibitor and Antifungal Agents against Aspergillus Species.

    PubMed

    Gao, Lujuan; Ding, Xiaozhen; Liu, Zhun; Wu, Qingzhi; Zeng, Tongxiang; Sun, Yi

    2016-06-01

    In vitro interactions of INK128, a target of rapamycin (TOR) kinase inhibitor, and antifungals, including itraconazole, voriconazole, posaconazole, amphotericin B, and caspofungin, against Aspergillus spp. were assessed with the broth microdilution checkerboard technique. Our results suggested synergistic effects between INK128 and all azoles tested, against multiple Aspergillus fumigatus and Aspergillus flavus isolates. However, no synergistic effects were observed when INK128 was combined with amphotericin B or caspofungin. No antagonism was observed for any combination. PMID:26976874

  3. Systemic movement of Aspergillus parasiticus in maize stalks and ears

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Preharvest infection of corn (Zea mays) kernels by Aspergillus flavus and A. parasiticus is a chronic problem in the southern United States. It has been reported that these fungi infect developing kernels via the silk. This study was conducted to explore other avenues of infection of corn ears by ...

  4. Biodiversity of Aspergillus section Flavi in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fungi belonging to Aspergillus section Flavi are of great economic importance in the United States due to the formation of toxic and carcinogenic aflatoxins in agricultural commodities. Development of control strategies against A. flavus and A. parasiticus, the major aflatoxin-producing species, is...

  5. Alginate Oligosaccharides Inhibit Fungal Cell Growth and Potentiate the Activity of Antifungals against Candida and Aspergillus spp

    PubMed Central

    Tøndervik, Anne; Sletta, Håvard; Klinkenberg, Geir; Emanuel, Charlotte; Powell, Lydia C.; Pritchard, Manon F.; Khan, Saira; Craine, Kieron M.; Onsøyen, Edvar; Rye, Phil D.; Wright, Chris; Thomas, David W.; Hill, Katja E.

    2014-01-01

    The oligosaccharide OligoG, an alginate derived from seaweed, has been shown to have anti-bacterial and anti-biofilm properties and potentiates the activity of selected antibiotics against multi-drug resistant bacteria. The ability of OligoG to perturb fungal growth and potentiate conventional antifungal agents was evaluated using a range of pathogenic fungal strains. Candida (n = 11) and Aspergillus (n = 3) spp. were tested using germ tube assays, LIVE/DEAD staining, scanning electron microscopy (SEM), atomic force microscopy (AFM) and high-throughput minimum inhibition concentration assays (MICs). In general, the strains tested showed a significant dose-dependent reduction in cell growth at ≥6% OligoG as measured by optical density (OD600; P<0.05). OligoG (>0.5%) also showed a significant inhibitory effect on hyphal growth in germ tube assays, although strain-dependent variations in efficacy were observed (P<0.05). SEM and AFM both showed that OligoG (≥2%) markedly disrupted fungal biofilm formation, both alone, and in combination with fluconazole. Cell surface roughness was also significantly increased by the combination treatment (P<0.001). High-throughput robotic MIC screening demonstrated the potentiating effects of OligoG (2, 6, 10%) with nystatin, amphotericin B, fluconazole, miconazole, voriconazole or terbinafine with the test strains. Potentiating effects were observed for the Aspergillus strains with all six antifungal agents, with an up to 16-fold (nystatin) reduction in MIC. Similarly, all the Candida spp. showed potentiation with nystatin (up to 16-fold) and fluconazole (up to 8-fold). These findings demonstrate the antifungal properties of OligoG and suggest a potential role in the management of fungal infections and possible reduction of antifungal toxicity. PMID:25409186

  6. The Gβ-like protein CpcB is required for hyphal growth, conidiophore morphology and pathogenicity in Aspergillus fumigatus.

    PubMed

    Cai, Zhen-dong; Chai, Yan-fei; Zhang, Cai-yun; Qiao, Wei-ran; Sang, Hong; Lu, Ling

    2015-08-01

    CpcB (cross pathway control B) encodes a yeast Cpc2 and mammalian RACK1 (receptor for activated protein kinase C) ortholog, which is a WD repeat protein with functional homology to the β subunit of heterotrimeric G proteins in Aspergillus fumigatus. Previous study has reported that CpcB governs growth and development in both A. fumigatus and Aspergillus nidulans. However, little is known about the functional identities of CpcB orthologs and their relationships with G protein complexes. In this study, we verified that cytoplasmic AfCpcB acts as a Gβ-like protein ortholog and plays important roles in hyphal growth, conidiophore morphology, cell wall integrity, and virulence in A. fumigatus. Furthermore, double deletion of AfcpcB and AfgpaB (Gα) causes a similar phenotype to AfgpaB mutant with abnormal multiple septa conidiophores but exhibits sparse conidiation with white and fluffy colonies. Thus, the exacerbated conidiation defect suggests that AfcpcB has its own specific function compared to the Gα subunit of AfgpaB or the G-protein complex. In addition, complementation assays using AfcpcB orthologs of A. nidulans and yeasts (Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida albicans) suggest that all tested fungal AfcpcB orthologs under the A. fumigatus native promoter can largely restore hyphal growth defects in AfcpcB deletion mutant, but only the A. nidulans cpcB ortholog completely rescues the ΔAfcpcB conidiation defect, suggesting that CpcB acts as a Gβ-like protein ortholog in the Aspergilli, but may have unique and important unexplored functions that required for conidiation, which is absent in yeast. PMID:25892048

  7. Expression profiling of non-aflatoxigenic Aspergillus parasiticus mutants obtained by 5-azacytosine treatment or serial mycelial transfer

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are carcinogenic secondary metabolites produced by the fungi Aspergillus flavus and Aspergillus parasiticus. Repeated serial mycelial transfer or treatment of A. parasiticus with 5-azacytidine produced mutants with a fluffy phenotype and loss of aflatoxin production. To understand how the...

  8. Relationships among resistances to Fusarium and Aspergillus ear rots and contamination by fumonisin and aflatoxin in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fusarium verticillioides, F. proliferatum, and Aspergillus flavus cause ear rots of maize and contaminate the grain with mycotoxins (fumonisin or aflatoxin). The objective of this study was to investigate the relationships between resistance to Fusarium and Aspergillus ear rots and fumonisin and a...

  9. Distribution and Toxigenicity of Aspergillus Species Isolated from Maize Kernels from Three Agro-ecological Zones in Nigeria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize samples were collected during a survey in three agro-ecological zones in Nigeria to determine the distribution and aflatoxin-producing potential of members of Aspergillus section Flavi. Among Aspergillus, A. flavus was the most predominant and L-strains constituted > 90% of the species identi...

  10. [Aspergillus spp. in risk areas of transplant patients in a university hospital].

    PubMed

    Cárdenas, María Ximena; Cortes, Jorge Alberto; Parra, Claudia Marcela

    2008-12-31

    As a consequence of the increase in the number of immunocompromised patients, cases of aspergillosis, due to the opportunist character of this fungus, have increased considerably. Aspergillus fumigatus, Aspergillus flavus, and Aspergillus niger have been found in air and water samples of the majority of investigated hospitals. The aim of the present study was to investigate the presence of aspergilli in transplant patients areas at the Hospital Universitario of Bogotá, Colombia. Samples of air were collected using the MAS-100 Air Sampler from each of the investigated areas. A sample of 100 ml of water was also recovered from these areas. All samples were taken for triplicate and were cultured in 2% Sabouraud Dextrose Agar. The average of aspergilli in air samples was 2.8 CFU/l corresponding to A. flavus, A. niger, Aspergillus versicolor and Aspergillus terreus. In water samples, the average was 17.1 CFU/l corresponding to A. flavus and Aspergillus clavatus. Because potentially pathogenic Aspergillus species were found in the hospital areas were transplant patients are usually kept, active surveillance and a high clinical suspicion should be considered in those patients. Since Aspergillus infections haven't been found so far, a higher fungal load and other host factors might be needed to facilitate the infection. PMID:19071892

  11. Lipase Production in Solid-State Fermentation Monitoring Biomass Growth of Aspergillus niger Using Digital Image Processing

    NASA Astrophysics Data System (ADS)

    Dutra, Julio C. V.; da Terzi, Selma C.; Bevilaqua, Juliana Vaz; Damaso, Mônica C. T.; Couri, Sônia; Langone, Marta A. P.; Senna, Lilian F.

    The aim of this study was to monitor the biomass growth of Aspergillus niger in solid-state fermentation (SSF) for lipase production using digital image processing technique. The strain A. niger 11T53A14 was cultivated in SSF using wheat bran as support, which was enriched with 0.91% (m/v) of ammonium sulfate. The addition of several vegetable oils (castor, soybean, olive, corn, and palm oils) was investigated to enhance lipase production. The maximum lipase activity was obtained using 2% (m/m) castor oil. In these conditions, the growth was evaluated each 24 h for 5 days by the glycosamine content analysis and digital image processing. Lipase activity was also determined. The results indicated that the digital image process technique can be used to monitor biomass growth in a SSF process and to correlate biomass growth and enzyme activity. In addition, the immobilized esterification lipase activity was determined for the butyl oleate synthesis, with and without 50% v/v hexane, resulting in 650 and 120 U/g, respectively. The enzyme was also used for transesterification of soybean oil and ethanol with maximum yield of 2.4%, after 30 min of reaction.

  12. Lipase production in solid-state fermentation monitoring biomass growth of aspergillus niger using digital image processing.

    PubMed

    Dutra, Júlio C V; da C Terzi, Selma; Bevilaqua, Juliana Vaz; Damaso, Mônica C T; Couri, Sônia; Langone, Marta A P; Senna, Lilian F

    2008-03-01

    The aim of this study was to monitor the biomass growth of Aspergillus niger in solid-state fermentation (SSF) for lipase production using digital image processing technique. The strain A. niger 11T53A14 was cultivated in SSF using wheat bran as support, which was enriched with 0.91% (m/v) of ammonium sulfate. The addition of several vegetable oils (castor, soybean, olive, corn, and palm oils) was investigated to enhance lipase production. The maximum lipase activity was obtained using 2% (m/m) castor oil. In these conditions, the growth was evaluated each 24 h for 5 days by the glycosamine content analysis and digital image processing. Lipase activity was also determined. The results indicated that the digital image process technique can be used to monitor biomass growth in a SSF process and to correlate biomass growth and enzyme activity. In addition, the immobilized esterification lipase activity was determined for the butyl oleate synthesis, with and without 50% v/v hexane, resulting in 650 and 120 U/g, respectively. The enzyme was also used for transesterification of soybean oil and ethanol with maximum yield of 2.4%, after 30 min of reaction. PMID:18401753

  13. Effects of Cymbopogon nardus (L.) W. Watson essential oil on the growth and morphogenesis of Aspergillus niger.

    PubMed

    de Billerbeck, V G; Roques, C G; Bessière, J M; Fonvieille, J L; Dargent, R

    2001-01-01

    The growth inhibitory effect of Cymbopogon nardus (L.) W. Watson var. nurdus essential oil on Aspergillus niger (Van Tieghem) mycelium was determined on agar medium. The mycelium growth was completely inhibited at 800 mg/L. This concentration was found to be lethal under the test conditions. Essential oil at 400 mg/L caused growth inhibition of 80% after 4 days of incubation, and a delay in conidiation of 4 days compared with the control. Microscopic observations were carried out to determine the ultrastructural modifications of A. niger hyphae after treatment with C. nardus essential oil. The main change observed by transmission electron microscopy concerned the hyphal diameter and the hyphal wall, which appeared markedly thinner. These modifications in cytological structure might be caused by the interference of the essential oil with the enzymes responsible for wall synthesis which disturb normal growth. Moreover, the essential oil caused plasma membrane disruption and mitochondrial structure disorganization. The findings thus indicate the possibility of exploiting Cymbopogon nardus essential oil as an effective inhibitor of biodegrading and storage-contaminating fungi. PMID:15049444

  14. The cell end marker protein TeaC is involved in growth directionality and septation in Aspergillus nidulans.

    PubMed

    Higashitsuji, Yuhei; Herrero, Saturnino; Takeshita, Norio; Fischer, Reinhard

    2009-07-01

    Polarized growth in filamentous fungi depends on the correct spatial organization of the microtubule (MT) and actin cytoskeleton. In Schizosaccharomyces pombe it was shown that the MT cytoskeleton is required for the delivery of so-called cell end marker proteins, e.g., Tea1 and Tea4, to the cell poles. Subsequently, these markers recruit several proteins required for polarized growth, e.g., a formin, which catalyzes actin cable formation. The latest results suggest that this machinery is conserved from fission yeast to Aspergillus nidulans. Here, we have characterized TeaC, a putative homologue of Tea4. Sequence identity between TeaC and Tea4 is only 12.5%, but they both share an SH3 domain in the N-terminal region. Deletion of teaC affected polarized growth and hyphal directionality. Whereas wild-type hyphae grow straight, hyphae of the mutant grow in a zig-zag way, similar to the hyphae of teaA deletion (tea1) strains. Some small, anucleate compartments were observed. Overexpression of teaC repressed septation and caused abnormal swelling of germinating conidia. In agreement with the two roles in polarized growth and in septation, TeaC localized to hyphal tips and to septa. TeaC interacted with the cell end marker protein TeaA at hyphal tips and with the formin SepA at hyphal tips and at septa. PMID:19429780

  15. Development in Aspergillus

    PubMed Central

    Krijgsheld, P.; Bleichrodt, R.; van Veluw, G.J.; Wang, F.; Müller, W.H.; Dijksterhuis, J.; Wösten, H.A.B.

    2013-01-01

    The genus Aspergillus represents a diverse group of fungi that are among the most abundant fungi in the world. Germination of a spore can lead to a vegetative mycelium that colonizes a substrate. The hyphae within the mycelium are highly heterogeneous with respect to gene expression, growth, and secretion. Aspergilli can reproduce both asexually and sexually. To this end, conidiophores and ascocarps are produced that form conidia and ascospores, respectively. This review describes the molecular mechanisms underlying growth and development of Aspergillus. PMID:23450714

  16. Effects of Cymbopogon citratus L. essential oil on the growth, lipid content and morphogenesis of Aspergillus niger ML2-strain.

    PubMed

    Helal, G A; Sarhan, M M; Abu Shahla, A N K; Abou El-Khair, E K

    2006-01-01

    The mycelial growth of Aspergillus niger van Tieghem was completely inhibited using 1.5 (microl/ml or 2.0 (microl/ml of Cymbopogon citratus essential oil applied by fumigation or contact method in Czapek liquid medium, respectively. This oil was found also to be fungicidal at the same concentrations. The sublethal doses 1.0 and 1.5 (microl/ml inhibited about 70% of fungal growth after five days of incubation and delayed conidiation as compared with the control. Microscopic observations using Light Microscope (LM), Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) were carried out to determine the ultra structural modifications of A. niger hyphae after treatment with C. citratus essential oil. The hyphal diameter and hyphal wall appeared markedly thinner. This oil also caused plasma membrane disruption and mitochondrial structure disorganization. Moreover, Ca+2, K+ and Mg+2 leakages increased from the fumigated mycelium and its total lipid content decreased, while the saturated fatty acids decreased and unsaturated fatty acids increased. These findings increase the possibility of exploiting C. citratus essential oil as an effective inhibitor of biodegrading and storage contaminating fungi and in fruit juice preservation. PMID:17139611

  17. Growth kinetics and mechanistic action of reactive oxygen species released by silver nanoparticles from Aspergillus niger on Escherichia coli.

    PubMed

    Ninganagouda, Shivaraj; Rathod, Vandana; Singh, Dattu; Hiremath, Jyoti; Singh, Ashish Kumar; Mathew, Jasmine; ul-Haq, Manzoor

    2014-01-01

    Silver Nanoparticles (AgNPs), the real silver bullet, are known to have good antibacterial properties against pathogenic microorganisms. In the present study AgNPs were prepared from extracellular filtrate of Aspergillus niger. Characterization of AgNPs by UV-Vis spectrum reveals specific surface plasmon resonance at peak 416 nm; TEM photographs revealed the size of the AgNPs to be 20-55 nm. Average diameter of the produced AgNPs was found to be 73 nm with a zeta potential that was -24 mV using Malvern Zetasizer. SEM micrographs showed AgNPs to be spherical with smooth morphology. EDS revealed the presence of pure metallic AgNPs along with carbon and oxygen signatures. Of the different concentrations (0, 2.5, 5, 10, and 15 μg/mL) used 10 μg/mL were sufficient to inhibit 10(7) CFU/mL of E. coli. ROS production was measured using DCFH-DA method and the the free radical generation effect of AgNPs on bacterial growth inhibition was investigated by ESR spectroscopy. This paper not only deals with the damage inflicted on microorganisms by AgNPs but also induces cell death through the production of ROS released by AgNPs and also growth kinetics of E. coli supplemented with AgNPs produced by A. niger. PMID:25028666

  18. Growth Kinetics and Mechanistic Action of Reactive Oxygen Species Released by Silver Nanoparticles from Aspergillus niger on Escherichia coli

    PubMed Central

    Ninganagouda, Shivaraj; Rathod, Vandana; Singh, Dattu; Hiremath, Jyoti; Singh, Ashish Kumar; Mathew, Jasmine; ul-Haq, Manzoor

    2014-01-01

    Silver Nanoparticles (AgNPs), the real silver bullet, are known to have good antibacterial properties against pathogenic microorganisms. In the present study AgNPs were prepared from extracellular filtrate of Aspergillus niger. Characterization of AgNPs by UV-Vis spectrum reveals specific surface plasmon resonance at peak 416 nm; TEM photographs revealed the size of the AgNPs to be 20–55 nm. Average diameter of the produced AgNPs was found to be 73 nm with a zeta potential that was −24 mV using Malvern Zetasizer. SEM micrographs showed AgNPs to be spherical with smooth morphology. EDS revealed the presence of pure metallic AgNPs along with carbon and oxygen signatures. Of the different concentrations (0, 2.5, 5, 10, and 15 μg/mL) used 10 μg/mL were sufficient to inhibit 107 CFU/mL of E. coli. ROS production was measured using DCFH-DA method and the the free radical generation effect of AgNPs on bacterial growth inhibition was investigated by ESR spectroscopy. This paper not only deals with the damage inflicted on microorganisms by AgNPs but also induces cell death through the production of ROS released by AgNPs and also growth kinetics of E. coli supplemented with AgNPs produced by A. niger. PMID:25028666

  19. IS CATALASE ACTIVITY ASSOCIATED WITH MAIZE RESISTANCE TO ASPERGILLUS FLAVUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Catalase activity was measured in various cob tissues during maize ear development because of its role in maintaining reactive oxygen homeostasis during biotic and abiotic stress. Catalase activity was determined in immature and mature embryos, pericarp, and rachis tissues of maize lines that are re...

  20. Submerged Conidiation and Product Formation by Aspergillus niger at Low Specific Growth Rates Are Affected in Aerial Developmental Mutants ▿

    PubMed Central

    Jørgensen, Thomas R.; Nielsen, Kristian F.; Arentshorst, Mark; Park, JooHae; van den Hondel, Cees A.; Frisvad, Jens C.; Ram, Arthur F.

    2011-01-01

    Exposure to an aerial environment or severe nutrient limitation induces asexual differentiation in filamentous fungi. Submerged cultivation of Aspergillus niger in carbon- and energy-limited retentostat cultures both induces and fuels conidiation. Physiological and transcriptomic analyses have revealed that this differentiation strongly affects product formation. Since conidiation is inherent in the aerial environment, we hypothesized that product formation near zero growth can be influenced by affecting differentiation or development of aerial hyphae in general. To investigate this idea, three developmental mutants (ΔfwnA, scl-1, and scl-2 mutants) that have no apparent vegetative growth defects were cultured in maltose-limited retentostat cultures. The secondary-metabolite profile of the wild-type strain defined flavasperone, aurasperone B, tensidol B, and two so far uncharacterized compounds as associated with conidium formation, while fumonisins B2, B4, and B6 were characteristic of early response to nutrient limitation by the vegetative mycelium. The developmental mutants responded differently to the severe substrate limitation, which resulted in distinct profiles of growth and product formation. fwnA encodes the polyketide synthase responsible for melanin biosynthesis during aerial differentiation, and we show that conidial melanin synthesis in submerged retentostat cultures and aurasperone B production are fwnA dependent. The scl-1 and scl-2 strains are two UV mutants generated in the ΔfwnA background that displayed reduced asexual conidiation and formed sclerotium-like structures on agar plates. The reduced conidiation phenotypes of the scl-1 and scl-2 strains are reflected in the retentostat cultivation and are accompanied by elimination or severely reduced accumulation of secondary metabolites and distinctly enhanced accumulation of extracellular protein. This investigation shows that submerged conidiation and product formation of a mitosporic fungus

  1. PINA is essential for growth and positively influences NIMA function in Aspergillus nidulans.

    PubMed

    Joseph, James D; Daigle, Scott N; Means, Anthony R

    2004-07-30

    The phospho-Ser/Thr-directed prolyl-isomerase Pin1 was originally identified in vertebrate systems as a negative regulator of NIMA, a Ser/Thr protein kinase that regulates the G(2)/M transition in Aspergillus nidulans. Here we explore the physiological roles of the Pin1 orthologue, PINA, in A. nidulans and evaluate the relevance of the interaction of PINA with NIMA in this fungus. We find pinA to be an essential gene in A. nidulans. In addition, when PINA levels are reduced 50-fold the cells grow at a reduced rate. Upon germination under conditions that repress PINA expression, the cells are delayed in the interphase activation of NIMX(cdc2), whereas they traverse the other phases of the cell cycle at a similar rate to controls. These results indicate that a marked reduction of PINA results in a lengthening of G(1). Additionally, PINA repression increases the rate at which the cells enter mitosis following release from a hydroxyurea arrest without altering the sensitivity of the fungus to agents that activate the replication or DNA damage checkpoints. In contrast to predictions based on Pin1, the physical interaction between PINA and NIMA is primarily dependent upon the prolylisomerase domain of PINA and the C-terminal 303 amino acids of NIMA. Finally, reduction of PINA levels exacerbates the nimA5 temperature-sensitive mutant, whereas overexpression of PINA decreases the severity of this mutation, results that are consistent with a positive genetic interaction between PINA and NIMA. Thus, although PINA is essential and positively regulates NIMA function, A. nidulans is most sensitive to a reduction in PINA concentration in G(1) rather than in G(2)/M. PMID:15178679

  2. Effect of water activity and temperature on the germination and growth of Aspergillus tamarii isolated from "Maldive fish".

    PubMed

    Mohamed, Shazla; Mo, Li; Flint, Steve; Palmer, Jon; Fletcher, Graham C

    2012-11-15

    Germination times and radial growth rates of cyclopiazonic acid producing strains of Aspergillus tamarii isolated from a smoked dried fish product were studied over water activities (a(w)) ranging from 0.99 to 0.79 at 25°C, 30°C, 35°C and 40°C on two laboratory media. The a(w) of the media was controlled by either NaCl or a mixture of glucose and fructose. The optimum germination and growth were observed at temperatures between 30°C and 35°C. Germination was favored at the highest a(w) of 0.99 under all conditions. Growth however was dependent on the media and temperature with a lower optimum a(w) of 0.95 for NaCl media and 0.95 to 0.92 a(w) on media containing glucose/fructose. The minimum a(w) for growth was often higher than for germination while both parameters were influenced by temperature and media type. Germination on NaCl media was prevented at a(w) values below 0.82 at 25°C and 30°C, 0.85 at 35°C and 40°C. However, growth did not occur at a(w) <0.85 at 25-35°C. At those temperatures on glucose/fructose media, growth was observed at the lowest a(w) tested (0.79). On both media, the restrictive effect of lowered water activity was more pronounced at 40°C than at 25-35°C. Delays in germination increased and growth rates decreased with marginal a(w) and temperature conditions. The fungi displayed better tolerance on glucose/fructose media than on NaCl media on which it was partly inhibited by the NaCl. The information obtained here could be used to develop strategies for the control of this xerophilic fungus on smoked dried fish and other tropical foods on which it predominates. PMID:23177051

  3. Effect of carbendazim and physicochemical factors on the growth and ochratoxin A production of Aspergillus carbonarius isolated from grapes.

    PubMed

    Medina, Angel; Mateo, Rufino; Valle-Algarra, Francisco M; Mateo, Eva M; Jiménez, Misericordia

    2007-11-01

    Carbendazim is a systemic fungicide that is commonly used on several crops (tobacco, fruit, vegetables, cereals, etc.). This fungicide is used to control fungal infections in vineyards. It is indicated against Botrytis cinerea, Uncinula necator, Plasmopara viticola and other fungi and can be used either alone or coupled with other fungicides. However, there is a lack of in-depth studies to evaluate its effectiveness against growth of Aspergillus carbonarius isolated from grapes and OTA production. A medium based on red grape juice was used in this study. Preliminary studies were performed at 0.98 a(w) and 25 degrees C using carbendazim concentrations over a wide range (1-2000 ng/ml medium) to control both growth of a strain of A. carbonarius isolated from grape and its ability to produce OTA. As the lag phase increased considerably at levels > 1000 ng/ml of medium, detailed studies were carried out in the range 50-450 ng/ml of medium at 0.98-0.94 a(w) and 20-28 degrees C. Statistical analysis (multifactor ANOVA) of the data revealed that the three factors assayed and the interactions a(w)-carbendazim concentration and a(w)-temperature had significant effects on lag phase duration. The highest lag-times were observed at 0.94 a(w,) 20 degrees C, and with 450 ng carbendazim/ml. The three factors also had significant effects of the growth rate and there was an interaction between a(w) and temperature. The growth rate of A. carbonarius in these cultures is favoured by high water availability and relatively high temperatures. However, addition of carbendazim at the assayed levels did not significantly influenced fungal growth rate. Accumulation of OTA was studied as a function of four factors (the three previously considered, and time). All factors had significant effects on the accumulation of OTA. There were also two significant interactions (a(w)-temperature and temperature-time). On the basis of the results obtained, carbendazim does not increase the lag phase of A

  4. Rapid Differentiation of Aspergillus Species from Other Medically Important Opportunistic Molds and Yeasts by PCR-Enzyme Immunoassay

    PubMed Central

    de Aguirre, Liliana; Hurst, Steven F.; Choi, Jong Soo; Shin, Jong Hee; Hinrikson, Hans Peter; Morrison, Christine J.

    2004-01-01

    We developed a PCR-based assay to differentiate medically important species of Aspergillus from one another and from other opportunistic molds and yeasts by employing universal, fungus-specific primers and DNA probes in an enzyme immunoassay format (PCR-EIA). Oligonucleotide probes, directed to the internal transcribed spacer 2 region of ribosomal DNA from Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus terreus, Aspergillus ustus, and Aspergillus versicolor, differentiated 41 isolates (3 to 9 each of the respective species; P < 0.001) in a PCR-EIA detection matrix and gave no false-positive reactions with 33 species of Acremonium, Exophiala, Candida, Fusarium, Mucor, Paecilomyces, Penicillium, Rhizopus, Scedosporium, Sporothrix, or other aspergilli tested. A single DNA probe to detect all seven of the most medically important Aspergillus species (A. flavus, A. fumigatus, A. nidulans, A. niger, A. terreus, A. ustus, and A. versicolor) was also designed. Identification of Aspergillus species was accomplished within a single day by the PCR-EIA, and as little as 0.5 pg of fungal DNA could be detected by this system. In addition, fungal DNA extracted from tissues of experimentally infected rabbits was successfully amplified and identified using the PCR-EIA system. This method is simple, rapid, and sensitive for the identification of medically important Aspergillus species and for their differentiation from other opportunistic fungi. PMID:15297489

  5. GROWTH EVALUATION OF FUNGI (PENICILLIUM AND ASPERGILLUS SPP.) ON CEILING TILES

    EPA Science Inventory

    The paper gives results of an evaluation of the potential for fungal growth on four different ceiling tiles in static chambers. It was found that even new ceiling tiles supported fungal growth under favorable conditions. Used ceiling tiles appeared to be more susceptible to funga...

  6. Comparative Secretome Analysis of Trichoderma reesei and Aspergillus niger during Growth on Sugarcane Biomass

    PubMed Central

    Borin, Gustavo Pagotto; Sanchez, Camila Cristina; de Souza, Amanda Pereira; de Santana, Eliane Silva; de Souza, Aline Tieppo; Leme, Adriana Franco Paes; Squina, Fabio Marcio; Buckeridge, Marcos; Goldman, Gustavo Henrique; Oliveira, Juliana Velasco de Castro

    2015-01-01

    Background Our dependence on fossil fuel sources and concern about the environment has generated a worldwide interest in establishing new sources of fuel and energy. Thus, the use of ethanol as a fuel is advantageous because it is an inexhaustible energy source and has minimal environmental impact. Currently, Brazil is the world's second largest producer of ethanol, which is produced from sugarcane juice fermentation. However, several studies suggest that Brazil could double its production per hectare by using sugarcane bagasse and straw, known as second-generation (2G) bioethanol. Nevertheless, the use of this biomass presents a challenge because the plant cell wall structure, which is composed of complex sugars (cellulose and hemicelluloses), must be broken down into fermentable sugar, such as glucose and xylose. To achieve this goal, several types of hydrolytic enzymes are necessary, and these enzymes represent the majority of the cost associated with 2G bioethanol processing. Reducing the cost of the saccharification process can be achieved via a comprehensive understanding of the hydrolytic mechanisms and enzyme secretion of polysaccharide-hydrolyzing microorganisms. In many natural habitats, several microorganisms degrade lignocellulosic biomass through a set of enzymes that act synergistically. In this study, two fungal species, Aspergillus niger and Trichoderma reesei, were grown on sugarcane biomass with two levels of cell wall complexity, culm in natura and pretreated bagasse. The production of enzymes related to biomass degradation was monitored using secretome analyses after 6, 12 and 24 hours. Concurrently, we analyzed the sugars in the supernatant. Results Analyzing the concentration of monosaccharides in the supernatant, we observed that both species are able to disassemble the polysaccharides of sugarcane cell walls since 6 hours post-inoculation. The sugars from the polysaccharides such as arabinoxylan and β-glucan (that compose the most external

  7. [Occurrence of indole alkaloids among secondary metabolites of soil Aspergillus].

    PubMed

    Vinokurova, N G; Khmel'nitskaia, I I; Baskunov, B P; Arinbasarov, M U

    2003-01-01

    The occurrence of indole alkaloids among secondary fungal metabolites was studied in species of the genus Aspergillus, isolated from soils that were sampled in various regions of Russia (a total of 102 isolates of the species A. niger, A. phoenicis, A. fumigatus, A. flavus, A. versicolor, A. ustus, A. clavatus, and A. ochraceus). Clavine alkaloids were represented by fumigaclavine, which was formed by A. fumigatus. alpha-Cyclopiazonic acid was formed by isolates of A. fumigatus, A. flavus, A. versicolor, A. phoenicis, and A. clavatus. The occurrence of indole-containing diketopiperazine alkaloids was documented for isolates of A. flavus, A. fumigatus, A. clavatus, and A. ochraceus. No indole-containing metabolites were found among the metabolites of A. ustus or A. niger. PMID:12722658

  8. The Weak Acid Preservative Sorbic Acid Inhibits Conidial Germination and Mycelial Growth of Aspergillus niger through Intracellular Acidification

    PubMed Central

    Plumridge, Andrew; Hesse, Stephan J. A.; Watson, Adrian J.; Lowe, Kenneth C.; Stratford, Malcolm; Archer, David B.

    2004-01-01

    The growth of the filamentous fungus Aspergillus niger, a common food spoilage organism, is inhibited by the weak acid preservative sorbic acid (trans-trans-2,4-hexadienoic acid). Conidia inoculated at 105/ml of medium showed a sorbic acid MIC of 4.5 mM at pH 4.0, whereas the MIC for the amount of mycelia at 24 h developed from the same spore inoculum was threefold lower. The MIC for conidia and, to a lesser extent, mycelia was shown to be dependent on the inoculum size. A. niger is capable of degrading sorbic acid, and this ability has consequences for food preservation strategies. The mechanism of action of sorbic acid was investigated using 31P nuclear magnetic resonance (NMR) spectroscopy. We show that a rapid decline in cytosolic pH (pHcyt) by more than 1 pH unit and a depression of vacuolar pH (pHvac) in A. niger occurs in the presence of sorbic acid. The pH gradient over the vacuole completely collapsed as a result of the decline in pHcyt. NMR spectra also revealed that sorbic acid (3.0 mM at pH 4.0) caused intracellular ATP pools and levels of sugar-phosphomonoesters and -phosphodiesters of A. niger mycelia to decrease dramatically, and they did not recover. The disruption of pH homeostasis by sorbic acid at concentrations below the MIC could account for the delay in spore germination and retardation of the onset of subsequent mycelial growth. PMID:15184150

  9. Inhibition of ochratoxin A production and growth of Aspergillus species by phenolic antioxidant compounds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The phenolic antioxidants, gallic acid, vanillic acid, protocatechuic acid, 4-hydroxybenzoic acid, catechin, caffeic acid, and chlorogenic acid were studied for their effects on ochratoxin A (OTA) production and fungal growth of ochratoxigenic Aspergilli. Of the 12 strains tested, which included A....

  10. Biodiversity of Aspergillus section Flavi in Europe in relation to the management of aflatoxin risk

    PubMed Central

    Perrone, Giancarlo; Gallo, Antonia; Logrieco, Antonio F.

    2014-01-01

    Aflatoxins and the producing fungi Aspergillus section Flavi are widely known as the most serious and dangerous mycotoxin issue in agricultural products. In Europe, before the outbreak of aflatoxins on maize (2003–2004) due to new climatic conditions, their contamination was confined to imported foods. Little information is available on molecular biodiversity and population structure of Aspergillus section Flavi in Europe. Preliminary reports evidenced the massive presence of Aspergillus flavus L -morphotype as the predominant species in maize field, no evidence of the highly toxigenic S-morphotype and of other aflatoxigenic species are reported. The risk of a shift in traditional occurrence areas for aflatoxins is expected in the world and in particular in South East of Europe due to the increasing average temperatures. Biological control of aflatoxin risk in the field by atoxigenic strains of A. flavus starts to be widely used in Africa and USA. Studies are necessary on the variation of aflatoxin production in populations of A. flavus to characterize stable atoxigenic A. flavus strains. The aim of present article is to give an overview on biodiversity and genetic variation of Aspergillus section Flavi in Europe in relation to the management of aflatoxins risk in the field. PMID:25101075

  11. Effect of stalk inoculation site on kernel infection of corn by Aspergillus parasiticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus and A. parasiticus may infect corn kernels on developing ears by several routes. Kernel infection is postulated to occur inside ears primarily by the colonization of silk tissues. Airborne conidia land on exposed silks, germinate, and then colonize the silks inside the ear. Onc...

  12. Understanding Nonaflatoxigenicity of Aspergillus sojae: A Windfall of Aflatoxin Biosynthesis Research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus section Flavi includes aflatoxin-producing and nonproducing fungi. A. sojae is unable to produce aflatoxins and is generally recognized as safe for food fermentation. However, because of its taxonomical relatedness to aflatoxin-producing A. parasiticus and A. flavus, it is necessary to...

  13. Evaluation of maize inbred lines for resistance to Aspergillus and Fusarium ear rot and mycotoxin accumulation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycotoxin contamination in corn grain is a worldwide threat to both human food safety and animal feed ingredients. A select group of inbred corn lines was evaluated in field trials for mycotoxin accumulation in grain and ear rot caused by Aspergillus flavus and Fusarium verticillioides. Our goal ...

  14. Functional and phylogenetic analysis of the Aspergillus ochraceoroseus aflQ (ordA) gene ortholog

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Within the Aspergillus parasiticus and A. flavus aflatoxin (AF) biosynthetic gene cluster the aflQ (ordA) and aflP (omtA) genes encode an oxidoreductase and methyltransferase, respectively. These genes are required for the final steps in the conversion of sterigmatocystin (ST) to aflatoxin B1 (AFB1...

  15. Inverse correlation of ability to produce aflatoxin and aspergillus colonization of maize seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seeds of aflatoxin-resistant and aflatoxin susceptible maize lines were inoculated with conidia of aflatoxin-producing Aspergillus flavus or A. parasiticus isolates or isogenic non-producing mutants. Conidial yields recovered from resistant maize seed after seven days were significantly lower for af...

  16. Deadly strains of Kenyan aspergillus are distinct from other aflatoxin producers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxin contamination of crops is a world-wide problem. Lethal aflatoxicosis of humans has been associated with maize produced in Kenya for over three decades. The S strain morphotype of Aspergillus flavus was identified as the primary cause of aflatoxin contamination events occurring between 2004...

  17. THE POPULATION GENETICS OF AFLATOXIN AND MATING TYPE EVOLUTION IN Aspergillus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins (AF) are toxic polyketides produced by several Aspergillus species that contaminate crops worldwide. A. parasiticus and A. flavus are the most common agents of AF contamination of corn, peanuts, cottonseed, figs and tree nuts in the US. The biosynthesis of AF involves over 20 enzymatic re...

  18. Polyketides from an Endophytic Aspergillus fumigatus Isolate Inhibit the Growth of Mycobacterium tuberculosis and MRSA.

    PubMed

    Flewelling, Andrew J; Bishop, Amanda I; Johnson, John A; Gray, Christopher A

    2015-10-01

    The crude extract of Aspergillusfumigatus isolate AF3-093A, an endophyte of the brown alga Fucus vesiculosus, showed significant antimicrobial activity in initial bioactivity screens. Bioassay-guided fractionation of the extract led to the isolation of flavipin, chaetoglobosin A and chaetoglobosin B, all of which inhibited the growth of Staphylococcus aureus, methicillin-resistant S. aureus and Mycobacterium tuberculosis H37Ra. The antimycobacterial activity of these compounds has not been previously reported. PMID:26669098

  19. Characterization of the bioactive metabolites from a plant growth-promoting rhizobacteria and their exploitation as antimicrobial and plant growth-promoting agents.

    PubMed

    George, Emrin; Kumar, S Nishanth; Jacob, Jubi; Bommasani, Bhaskara; Lankalapalli, Ravi S; Morang, P; Kumar, B S Dileep

    2015-05-01

    A plant growth-promoting bacterial strain, PM 105, isolated from a tea plantation soil from the North Eastern region of India was identified as Pseudomonas aeruginosa through classical and 16S ribosomal DNA (rDNA) gene sequencing. Further studies with this strain confirmed broad spectrum antifungal activity against ten human and plant pathogenic fungal pathogens viz. Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus tubingensis, Candida albicans, Colletotrichum gloeosporioides, Fusarium oxysporum, Pencillium expansum, Rhizoctonia solani, Trichophyton rubrum besides growth-promoting property in cowpea (Vigna unguiculata) and pigeon pea (Cajanus cajan). However, no antibacterial property was exhibited by this strain against the four test bacterial pathogens tested in agar overlay method. The crude bioactive metabolites produced by this strain were isolated with three different solvents that exhibited significant antimicrobial and plant growth-promoting activity. Chloroform extract recorded significant antimicrobial and plant growth-promoting activity. Three major compounds viz. 1-hydroxyphenazine, pyocyanin, and phenazine-1-carboxamide were purified and characterized from crude extracts of this strain by various spectral data. The purified compounds recorded prominent antimicrobial activity but failed to establish the plant growth promotion activity in test crop plants under gnotobiotic conditions. Pyocyanin recorded significant antimicrobial activity, and best activity was recorded against T. rubrum (29 mm), followed by P. expansum (28 mm). These results suggest the use of PM 105 as plant growth-promoting agent in crop plants after successful field trials. PMID:25832181

  20. Evaluation of antifungal activity of free fatty acids methyl esters fraction isolated from Algerian Linum usitatissimum L. seeds against toxigenic Aspergillus

    PubMed Central

    Abdelillah, Amrouche; Houcine, Benmehdi; Halima, Dalile; Meriem, Chabane sari; Imane, Zaaboub; Eddine, Smahi Djamal; Abdallah, Moussaoui; Daoudi, Chabane sari

    2013-01-01

    Objective The aim of this study was to evaluate the antifungal activity of the major fraction of fatty acids methyl esters (FAMEs) isolated from Linum usitatissimum L. seeds oil collected from Bechar department (Algeria). Methods The assessment of antifungal activity was carried out in terms of percentage of radial growth on solid medium (potatoes dextrose agar PDA) and biomass growth inhibition on liquid medium (potatoes dextrose broth PDB) against two fungi. Results The FAMEs was found to be effective in inhibiting the radial mycelial growth of Aspergillus flavus more than Aspergillus ochraceus on all tested concentrations. The highest antifungal index was found to be (54.19%) compared to Aspergillus ochraceus (40.48%). The results of the antifungal activity of the FAMEs inhibition of biomass on liquid medium gave no discounted results, but this does not exclude the antifungal activity. Conclusions We can assume that the observed antifungal potency may be due to the abundance of linoleic and α-linolenic acids in linseed oil which appears to be promising to treat fungal infections, storage fungi and food spoilage in food industry field. PMID:23730556