Concept: Aspergillus flavus
Aflatoxins (AFs) are highly carcinogenic compounds produced by Aspergillus species in seeds with high lipid and protein contents. It has been known for over 30 years that peptone is not conducive for AF productions, although reasons for this remain unknown.
Aflatoxin contamination in peanuts poses major challenges for vulnerable populations of sub-Saharan Africa and South Asia. Developing peanut varieties to combat pre-harvest Aspergillus flavus infection and resulting aflatoxin contamination has thus far remained a major challenge, confounded by highly complex peanut-Aspergilli pathosystem. Our study reports achieving high level of resistance in peanut by over expressing (OE) antifungal plant defensins MsDef1 and MtDef4.2, and through host-induced gene silencing (HIGS) of aflM and aflP genes from the aflatoxin biosynthetic pathway. While the former improves genetic resistance to A. flavus infection, the latter inhibits aflatoxin production in the event of infection providing durable resistance against different Aspergillus flavus morphotypes and negligible aflatoxin content in several peanut events/ lines well. A strong positive correlation was observed between aflatoxin accumulation and decline in transcription of the aflatoxin biosynthetic pathway genes in both OE-Def and HIGS lines. Transcriptomic signatures in the resistant lines revealed key mechanisms such as regulation of aflatoxin synthesis, its packaging and export control, besides the role of reactive oxygen species-scavenging enzymes that render enhanced protection in the OE and HIGS lines. This is the first study to demonstrate highly effective biotechnological strategies for successfully generating peanuts that are near-immune to aflatoxin contamination, offering a panacea for serious food safety, health and trade issues in the semi-arid regions. This article is protected by copyright. All rights reserved.
Aflatoxins are highly toxic, mutagenic, teratogenic and carcinogenic mycotoxins. Consumption of aflatoxin-contaminated food and commodities poses serious hazards to the health of humans and animals. Turmeric, Curcuma longa L., is a native plant of Southeast Asia and has antimicrobial, antioxidant and antifungal properties. This paper reports the antiaflatoxigenic activities of the essential oil of C. longa and curcumin. The medium tests were prepared with the oil of C. longa, and the curcumin standard at concentrations varied from 0.01% to 5.0%. All doses of the essential oil of the plant and the curcumin standard interfered with mycotoxin production. Both the essential oil and curcumin significantly inhibited the production of aflatoxins; the 0.5% level had a greater than 96% inhibitory effect. The levels of aflatoxin B(1) (AFB(1)) production were 1.0 and 42.7 μg/mL, respectively, for the samples treated with the essential oil of C. longa L. and curcumin at a concentration of 0.5%.
In recent years, Aspergillus species are reported frequently as aetiological agents of fungal keratitis in tropical countries such as India. Our aim was to evaluate the epidemiological features of Aspergillus keratitis cases over a 3-year period in a tertiary eye care hospital and to determine the antifungal susceptibilities of the causative agents. This study included culture proven Aspergillus keratitis cases diagnosed between September 2005 and August 2008. Data including prevalence, predisposing factors and demography were recorded, the isolates were identified by morphological and molecular methods and the minimum inhibitory concentration values of antifungal agents towards the isolates were determined by the microdilution method. Two hundred Aspergillus isolates were identified among 1737 culture proven cases. Most of the aspergilli (75%) proved to be A. flavus, followed by A. fumigatus (11.5%). Sixteen (8%) isolates belonged to species that are recently identified causative agents of mycotic keratitis. Most of the infected patients (88%) were adults ranging from 21 to 70 years of age. Co-existing ocular disease was confirmed in 16.5% of the patients. Econazole, clotrimazole and ketoconazole were notably active against A. flavus. Aspergillus keratitis is a significant problem in patients with ocular lesions in South-Indian States, warranting early diagnosis and initiation of specific antifungal therapy to improve outcome.
Aspergillosis is one of the most common causes of death in captive birds. Aspergillus fumigatus accounts for approximately 95 % of aspergillosis cases and Aspergillus flavus is the second most frequent organism associated with avian infections. In the present study, the fungi were grown from avian clinical samples (post-mortem lung material) and environmental samples (eggs, food and litter). Microsatellite markers were used to type seven clinical avian isolates and 22 environmental isolates of A. flavus. A. flavus was the only species (28 % prevalence) detected in the avian clinical isolates, whereas this species ranked third (19 %) after members of the genera Penicillium (39 %) and Cladosporium (21 %) in the environmental samples. Upon microsatellite analysis, five to eight distinct alleles were detected for each marker. The marker with the highest discriminatory power had eight alleles and a 0.852 D value. The combination of all six markers yielded a 0.991 D value with 25 distinct genotypes. One clinical avian isolate (lung biopsy) and one environmental isolate (egg) shared the same genotype. Microsatellite typing of A. flavus grown from avian and environmental samples displayed an excellent discriminatory power and 100 % reproducibility. This study showed a clustering of clinical and environmental isolates, which were clearly separated. Based upon these results, aspergillosis in birds may be induced by a great diversity of isolates.
Red rice is a fermented product of Monascus spp. It is widely consumed by Malaysian Chinese who believe in its pharmacological properties. The traditional method of red rice preparation disregards safety regulation and renders red rice susceptible to fungal infestation and mycotoxin contamination. A preliminary study was undertaken aiming to determine the occurrence of mycotoxigenic fungi and mycotoxins contamination on red rice at consumer level in Selangor, Malaysia. Fifty red rice samples were obtained and subjected to fungal isolation, enumeration, and identification. Citrinin, aflatoxin, and ochratoxin-A were quantitated by ELISA based on the presence of predominant causal fungi. Fungal loads of 1.4 × 10(4) to 2.1 × 10(6) CFU/g exceeded Malaysian limits. Monascus spp. as starter fungi were present in 50 samples (100 %), followed by Penicillium chrysogenum (62 %), Aspergillus niger (54 %), and Aspergillus flavus (44 %). Citrinin was present in 100 % samples (0.23-20.65 mg/kg), aflatoxin in 92 % samples (0.61-77.33 μg/kg) and Ochratoxin-A in 100 % samples (0.23-2.48 μg/kg); 100 % citrinin and 76.09 % aflatoxin exceeded Malaysian limits. The presence of mycotoxigenic fungi served as an indicator of mycotoxins contamination and might imply improper production, handling, transportation, and storage of red rice. Further confirmatory analysis (e.g., HPLC) is required to verify the mycotoxins level in red rice samples and to validate the safety status of red rice.
Aspergillus parasiticus produces the minor aflatoxins M(1) (AFM(1)), M(2) (AFM(2)), GM(1) (AFGM(1)), and GM(2) (AFGM(2)), as well as the major aflatoxins B(1) (AFB(1)), B(2) (AFB(2)), G(1) (AFG(1)), and G(2) (AFG(2)). Feeding of A. parasiticus with aspertoxin (12c-hydroxyOMST) caused AFM(1) and AFGM(1), and cell-free experiments using the microsomal fraction of A. parasiticus and aspertoxin caused production of AFM(1), indicating that aspertoxin is a precursor of AFM(1) and AFGM(1). Feeding of the same fungus with O-methylsterigmatocystin (OMST) caused AFM(1) and AFGM(1) together with AFB(1) and AFG(1); feeding with dihydroOMST (DHOMST) caused AFM(2) and AFGM(2) together with AFB(2) and AFG(2). Incubation of either the microsomal fraction or OrdA enzyme-expressing yeast with OMST caused production of aspertoxin together with AFM(1) and AFB(1). These results demonstrated that the OrdA enzyme catalyzes both 12c-hydroxylation reaction from OMST to aspertoxin and the successive reaction from aspertoxin to AFM(1). In contrast, feeding of the fungus with AFB(1) did not produce any AFM(1), demonstrating that M-/GM-aflatoxins are not produced from B-/G-aflatoxins. Furthermore, AFM(1) together with AFB(1) and AFG(1) was also produced from 11-hydroxyOMST (HOMST) in feeding experiment of A. parasiticus, whereas no aflatoxins were produced when used the ordA deletion mutant. These results demonstrated that OrdA enzyme can also catalyze 12c-hydroxylation of HOMST to produce 11-hydroxyaspertoxin, which serves as a precursor for the production of AFM(1) and AFGM(1). The same pathway may work for the production of AFM(2) and AFGM(2) from DHOMST and dihydroHOMST through the formation of dihydroaspertoxin and dihydro-11-hydroxyaspertoxin, respectively.
OBJECTIVE: The aim of the present study was to assess the anti-Aspergillus activity of culture filtrate of Streptomyces sp. VITSTK7 and biologically synthesized Ag2O/Ag nanoparticles using the culture filtrate of VITSTK7. MATERIAL AND METHODS: Silver nanoparticles were synthesized by biological reduction of silver nitrate using culture filtrate of Streptomyces sp. VITSTK7. The synthesized nanopaticles were characterized for surface plasma resonance peak, X-ray diffraction patterns, shape and size distribution. Inhibition of mycelial growth was used as an index of anti-Aspergillus activity of synthesized nanoparticles. RESULTS: The synthesized nanoparticles were spherical shaped and monodespersive in nature and showed a single surface plasma resonance peak at 420nm. X-ray diffraction patterns displayed typical peaks of crystalline silver oxide and size distribution histograms indicated production of 20-60-nm-size nanoparticles with average size of 35.2nm. The synthesized nanoparticles showed anti-Aspergillus activity against Aspergillus niger, Aspergillus flavus and Aspergillus fumigatus with antifungal index in the range of 62-75%. CONCLUSION: Thus the bioconversion of Ag2O/Ag nanoparticles by Streptomyces sp. VITSTK7 could be employed to control Aspergillus pathogenesis. The results of this study suggest that the green synthesis of Ag2O/Ag nanoparticles using marine Streptomyces sp. VITSTK7 yielded 27.9nm sized particles with potential to act against pathogenic Aspergillus species.
The aim of this study was to evaluate the effect of essential oil from fresh leaves of Sweet Fennel (Ocimum gratissimum) on mycoflora and Aspergillus section Flavi populations in stored peanuts. Aspergillus, Fusarium and Mucor spp. were the most common genera identified from peanuts at post-harvest in Benin by using a taxonomic schemes primarily based on morphological characters of mycelium and conidia. The isolated fungi include Aspergillus niger, A. parasiticus, A. flavus, A. ochraceus, Fusarium graminearum, F. solani, F. oxysporum and Mucor spp. The most prevalent fungi recorded were A. niger (94.18 %), A. flavus (83.72 %), A. parasiticus (77.90 %), A. ochraceus (72.09 %), F. graminearum (59.30 %) and F. oxysporum (51.16 %). Antifungal assay, performed by the agar medium assay, indicated that essential oil exhibited high antifungal activity against the growth of A. flavus, A. parasiticus, A. ochraceus and F. oxysporium. The minimal inhibitory concentration (MIC) of the essential oil was found to be 7.5 μl/ml for A. flavus and A. parasiticus and 5.5 μl/ml for A. ochraceus and F. oxysporium. The minimal fungicidal concentration (MFC) was recorded to be 8.0 μl/ml for A. flavus and A. parasiticus, 6,5 μl/ml for A. ochraceus and 6.0 μl/ml for F. oxysporium. The essential oil was found to be strongly fungicidal and inhibitory to aflatoxin production. Chemical analysis by GC/MS of the components of the oil led to the identification of 31 components characterized by myrcene (6.4 %), α-thujene (8.2 %), p-cymene (17.6 %), γ-terpinene (20.0 %), and thymol (26.9 %) as major components. The essential oil of Sweet Fennel, with fungal growth and mycotoxin inhibitory properties, offers a novel approach to the management of storage, thus opening up the possibility to prevent mold contamination in stored peanuts.
Efficacy of water dispersible formulations of biological control strains of Aspergillus flavus for aflatoxin management in corn
- Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment
- Published over 2 years ago
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 which was able to 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 21,882. Large sampling error, however limited the resolution of various treatment effects. Corn samples were also subjected to microbial analysis to better understand 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.