Specific Mycotoxin Research Articles

Emerging Alternaria toxins present in naturally contaminated dried tomatoes and their reduction using cold atmospheric plasma

This study investigated the efficacy of cold atmospheric plasma (CAP) treatment in degrading the emerging Alternaria toxins as pure molecules and as natural contaminants in dried tomatoes. The analyses of 25 batches of dried tomatoes showed that more than 40% of samples were contaminated with tenuazonic acid (TeA), one with Alternariol (AOH), while Tentoxin (TEN), Alternariol monomethyl ether (AME) and Altenuene (ALT) were not detected. CAP under ozone (O3) and NOx regimes were used to reduce the mycotoxin contamination, the results shown that O3 regime was more effective against all mycotoxins tested, while NOx regime showed different efficacy depending on the specific mycotoxin. The Weibull model proved to be appropriate for the degradation kinetics of all Alternaria toxins, regardless of the kinetic order they possess. Under the O3 regime, the resistance of the mycotoxin was AME > AOH > TEN > TeA > ALT. Our studies have shown for the first time that CAP reduces Alternaria toxins in naturally contaminated dried tomatoes. In particular, the rate of degradation under O3 regime increased significantly with the longest exposure time, reaching 52.80 ± 1.21 % of mycotoxin degradation. Our results expand our knowledge of CAP treatment as a potential means of reducing mycotoxin contamination in food. Further studies will be carried out to investigate the potential mycotoxin degradation products and to ensure food safety.

Prediction of Mycotoxin Response of DNA-Wrapped Nanotube Sensor with Machine Learning

DNA-wrapped single-walled carbon nanotubes (SWCNTs) have demonstrated great versatility in their use as optical sensors. SWCNTs emit a near-infrared fluorescence that is responsive to even the slightest changes in the nanotube environment, enabling sensors that can respond to single-molecule fluctuations within the vicinity of their surfaces. The fluorescence response and surface interactions of these sensors are determined by the DNA wrapping sequence. However, the lack of information on the relationship between the DNA sequence and its effect on the SWCNT fluorescence remains a bottleneck for designing sequences that are specific to analytes of interest. We have recently demonstrated the use of directed volution to control the fluorescence response of SWCNTs through DNA design. Iterative cycles of DNA mutation, screening, and selection allowed us to evolve sequences that yield DNA-wrapped SWCNT sensors with a desired fluorescence response to mycotoxins.In this work, we apply the screening results of the DNA libraries used in this approach to train machine learning (ML) algorithms. Artificial neural network (ANN) and support vector machine (SVM) methods were used to predict the response of ssDNA-SWCNT sensors to a specific mycotoxin. The reliability of these models was further assessed through cross-validation. The ANN and SVM models with cross-validation were able to accurately classify the various DNA sequences as yielding either a high or low fluorescence response with an accuracy of 73 and 81%, respectively. The models were further used to predict the performance of alternative DNA sequences outside the initial training dataset, using the Hierarchy and k-means++ clustering methods to evaluate the similarity and dissimilarity of each DNA sequence. Compared to the SVM model, the ANN model showed an improved ability to predict high responses for dissimilar DNA sequences. We further applied a combinatorial approach based on SVM and ANN models to design new DNA sequences for improving sensor performance. The success of this approach was validated experimentally, demonstrating the rational design of improved sensors with 95% prediction accuracy.The application of ML algorithms to directed evolution libraries of DNA thus allows one to accurately map the performances of these sensors within a particular sequence space. The computational success of this mapping provides a framework for replacing current empirical approaches with the rational design of DNA sequences for SWCNT sensing. Keywords: Nano Biosensor, DNA-SWCNT, Mycotoxin, Machine learning, Directed evolution method.

Prediction of Mycotoxin Response of DNA-Wrapped Nanotube Sensor with Machine Learning

DNA-wrapped single-walled carbon nanotubes (SWCNTs) have demonstrated great versatility in their use as optical sensors. SWCNTs emit a near-infrared fluorescence that is responsive to even the slightest changes in the nanotube environment, enabling sensors that can respond to single-molecule fluctuations within the vicinity of their surfaces. The fluorescence response and surface interactions of these sensors are determined by the DNA wrapping sequence. However, the lack of information on the relationship between the DNA sequence and its effect on the SWCNT fluorescence remains a bottleneck for designing sequences that are specific to analytes of interest. We have recently demonstrated the use of directed evolution to control the fluorescence response of SWCNTs through DNA design. Iterative cycles of DNA mutation, screening, and selection allowed us to evolve sequences that yield DNA-wrapped SWCNT sensors with a desired fluorescence response to mycotoxins.In this work, we apply the screening results of the DNA libraries used in this approach to train machine learning (ML) algorithms. Artificial neural network (ANN) and support vector machine (SVM) methods were used to predict the response of ssDNA-SWCNT sensors to a specific mycotoxin. The reliability of these models was further assessed through cross-validation. The ANN and SVM models with cross-validation were able to accurately classify the various DNA sequences as yielding either a high or low fluorescence response with an accuracy of 73 and 81%, respectively. The models were further used to predict the performance of alternative DNA sequences outside the initial training dataset, using the Hierarchy and k-means ++ clustering methods to evaluate the similarity and dissimilarity of each DNA sequence. Compared to the SVM model, the ANN model showed an improved ability to predict high responses for dissimilar DNA sequences. We further applied a combinatorial approach based on SVM and ANN models to design new DNA sequences for improving sensor performance. The success of this approach was validated experimentally, demonstrating the rational design of improved sensors with 95% prediction accuracy.The application of ML algorithms to directed evolution libraries of DNA thus allows one to accurately map the performances of these sensors within a particular sequence space. The computational success of this mapping provides a framework for replacing current empirical approaches with the rational design of DNA sequences for SWCNT sensing. Keywords: Nano Biosensor, DNA-SWCNT, Mycotoxin, Machine learning, Directed evolution method

Early detection of quality alterations in corn grains stored in vertical prototype silos using real-time monitoring of carbon dioxide and equilibrium moisture content

Early detection of the deterioration of grain stored in vertical silos would benefit food safety in these systems. Real-time monitoring of the intergranular variables - temperature, relative humidity, carbon dioxide and moisture content – was used to determine equilibrium moisture content and dry matter loss in corn stored in vertical prototype silos. Quantitative relationships were established for changes in quality variables – germination rate, apparent specific mass, starch content, protein solubility, pasting metrics, lipid activity and mycotoxin contamination. The results demonstrate that integrated monitoring of the equilibrium moisture content and carbon dioxide concentration in the stored corn mass can inform decision making for ideal storage time, loss reduction and food safety.

Aptamer-engineered nanomaterials to aid in mycotoxin determination

Mycotoxin prevalence in food is a global public health issue, that could impose severe economic burdens. To prevent risk of exposure to mycotoxins, it is of great importance to develop selective, sensitive, and reliable methods to monitor mycotoxin levels in food samples. Recent advances in nanotechnology have produced many nanomaterials with unique physical features and extensive effort has been exerted to apply these nanomaterials in mycotoxin determination. Many specific aptamers, which have distinct advantages such as uniform synthesis, low cost, customizable modification, and chemical stability, have been developed to endow nanomaterials with target selective properties. Given the merits of these materials, aptamer-based nanomaterials for mycotoxin detection have received extensive attention from researchers. This review focuses on recent advances in aptamer nanomaterial-based sensing strategies used for mycotoxin analysis in food samples in the past five years. Representative examples of aptamer nanomaterial-based sensors for specific mycotoxin detection are described. The future challenges and opportunities regarding the use of aptamer-based nanomaterials to assist in the control of mycotoxins and other food contaminants are also discussed.

Simultaneous Quantification of Aflatoxin B1, T-2 Toxin, Ochratoxin A and Deoxynivalenol in Dried Seafood Products by LC-MS/MS.

Mycotoxins are secondary metabolites produced by fungi. These contaminate dried seafoods during processing and storage and represent a potential health hazard for consumers. A sensitive, selective and accurate liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was established for simultaneous quantification of four common mycotoxins (aflatoxin B1 (AFB1), T-2 toxin (T-2), ochratoxin A (OTA) and deoxynivalenol (DON)) in dried shrimp, dried fish and dried mussel products. Mycotoxins were extracted from dried seafood samples by acetonitrile/water (85/15, v/v), subjected to ultrasound for 60 min at 20 °C and cleaned up by defatting with n-hexane. The sample matrix affected the linearity of detection (R2 ≥ 0.9974). The limit of detection (LOD) and limit of quantification (LOQ) in dried seafood products varied from 0.1 to 2.0 µg·kg−1 and 0.3 to 5.0 µg·kg−1, respectively. The method was validated by spiking samples with specific mycotoxin levels, and the recoveries, intra-relative standard deviation (RSDs) and inter-RSDs ranged between 72.2–98.4%, 2.8–10.6%, and 5.5–15.4%, respectively. This method was used to analyze 40 dried seafood products purchased from the Zhanjiang seafood market. Results of this product sampling showed that while no DON was detected, AFB1, T-2 and OTA were detected in 30.8%, 17.5% and 33.3% of the samples, respectively. AFB1, T-2 and OTA concentrations varied at 0.58–0.89, 0.55–1.34 and 0.36–1.51 µg·kg−1, respectively. Relatively high frequency of contamination and the presence of AFB1, OTA and T-2 residues indicate the need to monitor mycotoxins in dried seafood products.

Molecular and metabolic strategies for postharvest detection of heat-resistant fungus Neosartorya fischeri and its discrimination from Aspergillus fumigatus

Abstract Heat Resistant Fungi (HRF) and toxigenic fungi are considered as a serious problem both in the agricultural field and for human health, due to ascospores and mycotoxins production, which can contaminate fruit and, as a consequence, adversely affect the agricultural food chain. One strategy to identify these fungi is the use of modern molecular method, which include the analysis of DNA target regions for differentiation of the fungal species. However, previously developed methods included only the identification of pure strains but not the detection of Neosartorya fischeri in artificially contaminated food samples, such as fruit or juices. Therefore, the aim of presented study was to develop a detection method of Neosartorya fischeri in contaminated strawberry fruit and juice. The other strategy is the use of phenotypic assays to determine the metabolic profile of the fungi in order to facilitate a qualitative and quantitative detection of microorganisms based on specific substrates utilization and mycotoxins production. Accordingly, the study included an evaluation of the differences in phenotype profile between N. fischeri and the phylogenetically close fungus Aspergillus fumigatus, as a strategy of their differentiation and identification. PCR detection assay was developed that revealed the presence of N. fischeri DNA in all tested contaminated samples of fruit and juice. Therefore, it can be concluded that this rapid molecular method is an important tool for the evaluation of the postharvest quality of agricultural raw materials. Moreover, the results suggest that specific metabolic and mycotoxin patterns may be used as N. fischeri detection markers and strategy in discrimination of this fungus from A. fumigatus. The results indicated that N. fischeri and A. fumigatus had a different time period of carbon sources utilization, and particularly N. fischeri presented a more efficient carbon metabolism. Mycotoxins, verruculogen and fumitremorgin C, were detected after 4 days incubation of N. fischeri. Although metabolic assays are not such fast as molecular detection approach, they allow to deeper insight into the pathways activated by heat-resistant and toxigenic fungi. Therefore, both molecular and metabolic strategies of heat-resistant fungus detection and identification are complementary and can be used to measure postharvest quality of fruit and their products.

Mycotoxins in aquaculture: feed and food

AbstractMycotoxins, secondary metabolites produced by moulds, are responsible for causing significant economic losses due to spoilage of agricultural products but also due to direct or indirect health impact on livestock upon ingestion of mycotoxin contaminated feedstuffs. Aquaculture farmed species are not an exception and studies reporting mycotoxin‐related issues in the aquaculture industry have been increasing. However, our understanding on the prevalence and impact of mycotoxins in the aquaculture sector is still lower compared to the terrestrial livestock sector. Consequently, regulatory limits and guidance values have been defined based on the studies on terrestrial farm animals. The aim of this review is to compile and critically assess mycotoxin occurrence and co‐occurrence in aquaculture finished feeds, and understand the risk of mycotoxin carry‐over in aquaculture seafood products. Furthermore, we aim with this review to raise awareness to the scientific community, the regulatory authorities and the aquaculture industry to the need for specific aquaculture mycotoxin maximum concentration levels for both aquaculture feeds and foods.

In vitro model to assess the adsorption of oral veterinary drugs to mycotoxin binders in a feed- and aflatoxin B1-containing buffered matrix

ABSTRACTMycotoxin binders are feed additives which are mixed in the feed to adsorb mycotoxins and thereby reducing their toxic effects on animals. Interactions with orally administered veterinary medicinal products, such as antimicrobials or coccidiostats, have been reported previously. This paper describes an in vitro model to screen the interaction between mycotoxin binders and veterinary drugs with respect to the non-specific binding of drugs. It is designed as a static setup using a single concentration of drug and binder in a feed-containing or a feed-plus-mycotoxin-containing matrix, buffered at different pH values. The model was applied to two frequently used antimicrobials in veterinary medicine, doxycycline (DOX) and tylosin (TYL), one major mycotoxin, aflatoxin B1 (AFB1), and four mycotoxin binders. Proportions of feed, DOX or TYL, AFB1, and binder are equivalent to the in vivo situation for broiler chickens, while pH and volume of the buffer are representative of the gastrointestinal tract of chickens. A substantial binding of DOX (~ 88%) and TYL (~ 66%) to the feed-matrix was observed. For the mycotoxin binders, similar results were obtained for DOX and TYL; more specifically up to an inclusion rate of 20 g binder/kg feed, no significant binding was demonstrated, determined as the free concentration of DOX and TYL. A single exception was noticed for TYL and one specific bentonite-based mycotoxin binder, for which no significant interaction could be demonstrated up to 10 g binder/kg but there was an effect at 20 g/kg. In all cases, there was no competition between the tested drugs DOX or TYL and the mycotoxin AFB1 for binding to the bentonite-based mycotoxin binder.

Mycotoxin monitoring for commercial foodstuffs in Taiwan

Mycotoxins are toxic food contaminants that are naturally produced by certain fungi. They induce negative effects on human health by making food unsafe for consumption. In this study, analyses were performed to determine the levels and incidence of aflatoxins (AFs) in peanut products, tree nuts, spices, and Coix seeds; ochratoxin A (OTA) in wheat and roasted coffee, as well as OTA and AFs in rice; and citrinin (CIT) in red yeast rice (RYR) products. A total of 712 samples from nine different food categories were collected between 2012 and 2013. The samples were analyzed over 2 years for AFs, OTA, and CIT by methods recommended by the Ministry of Health and Welfare. These official analytical methods were extensively validated in-house and through interlaboratory trials. The analytical values of suspected contaminated specimens were confirmed by liquid chromatography – tandem mass spectrometry analysis to identify the specific mycotoxin present in the sample. We show that 689 samples (96.8%) complied with the regulations set by the Ministry of Health and Welfare. AFs were found in four peanut-candy products, one peanut-flour product, one pistachio product, one Sichuan-pepper product, and one Coix seed product. All had exceeded the maximum levels of 15 parts per billion for peanut and 10 parts per billion for other food products. Furthermore, 14 RYR samples contained CIT above 5 parts per million, and one RYR tablet exceeded the maximum amount allowed. Instances of AFs in substandard Sichuan pepper and Coix seeds were first detected in Taiwan. Measures were taken by the relevant authorities to remove substandard products from the market in order to decrease consumer exposure to mycotoxin. Border control measures were applied to importing food commodities with a higher risk of mycotoxin contamination, such as peanut, Sichuan pepper, and RYR products. Declining trends were observed in the noncompliance rate of AFs in peanut products, as well as that of CIT in RYR raw materials monitored from 2010 to 2013.

Environmental mold and mycotoxin exposures elicit specific cytokine and chemokine responses.

BackgroundMolds can cause respiratory symptoms and asthma. We sought to use isolated peripheral blood mononuclear cells (PBMCs) to understand changes in cytokine and chemokine levels in response to mold and mycotoxin exposures and to link these levels with respiratory symptoms in humans. We did this by utilizing an ex vivo assay approach to differentiate mold-exposed patients and unexposed controls. While circulating plasma chemokine and cytokine levels from these two groups might be similar, we hypothesized that by challenging their isolated white blood cells with mold or mold extracts, we would see a differential chemokine and cytokine release.Methods and FindingsPeripheral blood mononuclear cells (PBMCs) were isolated from blood from 33 patients with a history of mold exposures and from 17 controls. Cultured PBMCs were incubated with the most prominent Stachybotrys chartarum mycotoxin, satratoxin G, or with aqueous mold extract, ionomycin, or media, each with or without PMA. Additional PBMCs were exposed to spores of Aspergillus niger, Cladosporium herbarum and Penicillium chrysogenum. After 18 hours, cytokines and chemokines released into the culture medium were measured by multiplex assay. Clinical histories, physical examinations and pulmonary function tests were also conducted. After ex vivo PBMC exposures to molds or mycotoxins, the chemokine and cytokine profiles from patients with a history of mold exposure were significantly different from those of unexposed controls. In contrast, biomarker profiles from cells exposed to media alone showed no difference between the patients and controls.ConclusionsThese findings demonstrate that chronic mold exposures induced changes in inflammatory and immune system responses to specific mold and mycotoxin challenges. These responses can differentiate mold-exposed patients from unexposed controls. This strategy may be a powerful approach to document immune system responsiveness to molds and other inflammation-inducing environmental agents.

Fungal metabolites diversity in maize and associated human dietary exposures relate to micro-climatic patterns in Malawi

This study investigated the diversity of fungal metabolites in maize across four agro-ecological zones of Malawi. A total of 90 maize samples (for human consumption), collected from farmsteads, were analysed for 235 fungal metabolites using liquid chromatography-tandem mass spectrometry. A total of 65 metabolites were found in the samples. 75% of samples from the hottest agro-ecological zone contained either aflatoxins, fumonisins, deoxynivalenol, zearalenone; or a combination thereof in levels exceeding European Union (EU) maximum levels, whereas the related fraction was only 17% in the cool temperature zone. Aflatoxins, citrinin, 3-nitropropionic acid, monocerin and equisetin were most prevalent and in higher levels in samples from hot agro-ecological zones, whereas deoxynivalenol, nivalenol, zearalenone and aurofusarin were most prevalent in cool agro-ecologies. On the basis of per-capita maize consumption, estimated daily intakes for all samples from hot ecologies were well above the JECFA's provisional maximum tolerable daily intake (PMTDI) of 2.0 μg/kg body weight (bw)/day for fumonisins, whereas the PMTDI of 1.0 μg/kg bw/day for deoxynivalenol was exceeded in relatively more (90%) samples from the cool highlands than the other zones. These results demonstrate the influence of micro-climatic conditions on mycotoxin prevalence patterns and underscores the need for development of agro-ecological specific mycotoxin dietary exposure management strategies.

Multitoxin analysis of Aspergillus clavatus-infected feed samples implicated in two outbreaks of neuromycotoxicosis in cattle in South Africa.

Aspergillus clavatus intoxication is a highly fatal neuromycotoxicosis of ruminants, especially cattle. It is caused by the ingestion of infected sprouting grain and sorghum beer residue. Locomotor disturbances, tremors and paralysis are observed. Histologically, degeneration and necrosis of larger neurons in the medulla oblongata, the midbrain, the thalamus and the ventral horns of the spinal cord are observed. Although a range of mycotoxins such as patulin, cytochalasin E and pseurotin A have been isolated, there is limited information on which specific mycotoxin or group of mycotoxins are involved during outbreaks of intoxication in livestock. In the present study, two outbreaks of A. clavatus poisoning in cattle are briefly described. Feed samples were collected for fungal identification, and culture and multitoxin analysis. A range of fungal metabolites were detected, and the estimated concentrations (μg/kg) are provided. Both the sprouting barley and brewer's grain were predominantly infected with A. clavatus and, to a lesser extent, Rhizopus arrhizus. The only common Aspergillus secondary metabolite present in all the samples was pseurotin A. Patulin and cytochalasin E were present in the sprouting barley samples, as well as the A. clavatus isolates cultured on malt extract agar for 2 weeks; however, neither of these mycotoxins could be detected in the brewer's grain sample.

Near- and mid-infrared spectroscopy as efficient tools for detection of fungal and mycotoxin contamination in agricultural commodities

In the last decade, near-infrared (NIR) and mid-infrared (MIR) spectroscopy have proven to be most promising tools for the detection of fungal contamination and estimation of mycotoxins in agricultural commodities, particularly of cereals. Owing to significant economic losses incurred from fungal contamination of foodstuffs, producers and processors are looking for fast, reliable, and less-expensive methods for the detection of fungal damage. In this context, NIR and MIR spectroscopy offer a fast, less-expensive, non-destructive, and relatively simple analytical method. Results from published studies indicate that NIR and MIR spectroscopy can be successfully applied to identifying fungal contamination and estimating specific mycotoxins. This review will focus on the applications of NIR and MIR spectroscopy to the classification of fungal contamination and the determination of specific mycotoxin contamination levels, and to compare this technology with traditional analytical methods.

Label-free homogeneous FRET immunoassay for the detection of mycotoxins that utilizes quenching of the intrinsic fluorescence ofantibodies

The phenomenon of fluorescence quenching of an antibody by a specific ligand was applied in developing a technique for detection of mycotoxins, such as aflatoxin B1 (AFB1), ochratoxin A, and zearalenone. Studies showed that the intrinsic fluorescence of tryptophan (Trp) residues in antibodies, promoted by excitation at 280nm, is quenched upon binding of specific mycotoxin ligands. Fluorescence quenching in FRET system takes place in these systems as a consequence of the overlap of the emission spectra of antibody donors with the absorption spectra of the mycotoxins. Further studies focusing on the detection of AFB1 revealed that the Fab fragment, the variable region of the antibody where specific binding of AFB1 occurs, can be utilized to increase the sensitivity of the detection system. The results demonstrate that fluorescence of the Fab fragment is almost completely quenched by AFB1 whereas emission from intact anti-AFB1 is only partially quenched by this mycotoxin. The limits of detection (LODs) were found to be 0.85 and 0.09ngmL−1 for assays using the intact antibody and the Fab fragment, respectively, corresponding to a 10-fold enhancement. A practical application of the Fab fragment based assay system was demonstrated by its use in the detection of AFB1 in spiked barley grain samples. The observations made in this effort show that the new, label-free, non-competitive, and homogeneous FRET immunoassay strategy, which requires a simple sample preparation procedure, serves as a powerful tool for the rapid and sensitive quantitative determination of organic substances such as mycotoxin.

Genotyping and phenotyping of Fusarium graminearum isolates from Germany related to their mycotoxin biosynthesis

Fusarium graminearum is the most important pathogen causing Fusarium head blight (FHB) of small cereal grains worldwide responsible for quantitative and qualitative yield losses. The presence in crops is often associated with mycotoxin contamination of foodstuff limiting its use for human and animal consumption. A collection of isolates of F. graminearum from Germany was characterized genetically and chemically for their potential to produce the B trichothecenes deoxynivalenol (DON) and nivalenol (NIV). Molecular methods with eight PCR assays were implemented based on functional Tri7 and Tri13 genes and on the tri5-tri6 intergenic region to differentiate between chemotaxonomic groups DON and NIV, resulting in a marked majority (61/63) of DON chemotypes. Mycotoxins produced on rice kernels were quantified by means of LC-MSMS including DON, NIV, 3-acetyl-DON (3-ADON), 15-acetyl-DON (15-ADON), DON-3-glucoside, fusarenon X, as well as zearalenone; all of them proving to be present in high concentration among the isolates. All DON-chemotype isolates also produced lower amounts of NIV with the amount being positively correlated (R²=0.89) to the DON amount. 15-ADON and 3-ADON are reported to be produced simultaneously by the isolates, the former dominating over the latter in all but one isolate. Fungal biomass, was quantified via ergosterol amount on rice. It was used to calculate specific mycotoxin production per biomass of isolates, ranging from 0.104 to 1.815mg DON mg-1 ergosterol, presenting a Gaussian distribution. Genotype and phenotype characterization revealed discrepancies with respect to mycotoxin production potential of the fungi, i.e. isolates from one chemotype were able to produce mycotoxins from other chemotypes in considerable amounts.

Quantification of Fusarium poae DNA and associated mycotoxins in asymptomatically contaminated wheat

Recent surveys have identified increased predominance of Fusarium poae causing FHB ( Fusarium Head Blight) of wheat in Europe. Several studies revealed a correlation between levels of F. poae DNA and nivalenol (NIV) and enniatins (ENNs) in highly contaminated cereal grain. In this study, F. poae specific TaqMan assays and mycotoxin analysis were performed on 48 asymptomatically contaminated wheat grain samples obtained from six different locations in northern Poland in 2006 and 2007. TaqMan assays revealed the presence of F. poae DNA in all samples analyzed, however the amounts of target DNA between the samples differed. Mycotoxin analysis revealed the presence of 13 toxins in the grain analyzed, however only ENN B and B1 were detected at quantifiable concentrations. A significant positive correlation was revealed between F. poae DNA ( R = 0.75) and monthly mean rainfalls recorded in May (a month before wheat anthesis) in both years. Additionally, a significant positive correlation was found between levels of ENN B+B1 ( R = 0.49) and rainfalls in May, however, no correlation was found between the quantity F. poae DNA and the level of ENN B+B1. This paper confirms the increasing importance of F. poae in the FHB complex of wheat in Poland.

Transcriptional regulation of chemical diversity in Aspergillus fumigatus by LaeA.

Secondary metabolites, including toxins and melanins, have been implicated as virulence attributes in invasive aspergillosis. Although not definitively proved, this supposition is supported by the decreased virulence of an Aspergillus fumigatus strain, ΔlaeA, that is crippled in the production of numerous secondary metabolites. However, loss of a single LaeA-regulated toxin, gliotoxin, did not recapitulate the hypovirulent ΔlaeA pathotype, thus implicating other toxins whose production is governed by LaeA. Toward this end, a whole-genome comparison of the transcriptional profile of wild-type, ΔlaeA, and complemented control strains showed that genes in 13 of 22 secondary metabolite gene clusters, including several A. fumigatus–specific mycotoxin clusters, were expressed at significantly lower levels in the ΔlaeA mutant. LaeA influences the expression of at least 9.5% of the genome (943 of 9,626 genes in A. fumigatus) but positively controls expression of 20% to 40% of major classes of secondary metabolite biosynthesis genes such as nonribosomal peptide synthetases (NRPSs), polyketide synthases, and P450 monooxygenases. Tight regulation of NRPS-encoding genes was highlighted by quantitative real-time reverse-transcription PCR analysis. In addition, expression of a putative siderophore biosynthesis NRPS (NRPS2/sidE) was greatly reduced in the ΔlaeA mutant in comparison to controls under inducing iron-deficient conditions. Comparative genomic analysis showed that A. fumigatus secondary metabolite gene clusters constitute evolutionarily diverse regions that may be important for niche adaptation and virulence attributes. Our findings suggest that LaeA is a novel target for comprehensive modification of chemical diversity and pathogenicity.

The medical effects of mold exposure

Exposure to molds can cause human disease through several well-defined mechanisms. In addition, many new mold-related illnesses have been hypothesized in recent years that remain largely or completely unproved. Concerns about mold exposure and its effects are so common that all health care providers, particularly allergists and immunologists, are frequently faced with issues regarding these real and asserted mold-related illnesses. The purpose of this position paper is to provide a state-of-the-art review of the role that molds are known to play in human disease, including asthma, allergic rhinitis, allergic bronchopulmonary aspergillosis, sinusitis, and hypersensitivity pneumonitis. In addition, other purported mold-related illnesses and the data that currently exist to support them are carefully reviewed, as are the currently available approaches for the evaluation of both patients and the environment.

Saliva Secretory IgA Antibodies Against Molds and Mycotoxins in Patients Exposed to Toxigenic Fungi

Upper respiratory exposure to different environmental antigens results first in the activation of mucosal immunity and production of IgA antibodies in different secretions including saliva. Despite this there is no study, which addresses secretory antibodies against molds and mycotoxins. The purpose of this study was to evaluate mold‐specific salivary IgA in individuals exposed to molds and mycotoxins in a water‐damaged building environment. Saliva IgA antibody levels against seven different molds and two mycotoxins were studied in 40 patients exposed to molds and in 40 control subjects. Mold‐exposed patients showed significantly higher levels of salivary IgA antibodies against one or more mold species. A majority of patients with high IgA antibodies against molds exhibited elevation in salivary IgA against mycotoxins, as well. These IgA antibodies against molds and mycotoxins are specific, since using molds and mycotoxins in immune absorption could reduce antibody levels, significantly. Detection of high counts of molds in water‐damaged buildings, strongly suggests the existence of a reservoir of mold spores in the environment. This viable microbial activity with specific mold and mycotoxin IgA in saliva may assist in the diagnosis of mold exposure. Whether mold and mycotoxin specific IgA antibodies detected in saliva are indicative of the role of IgA antibodies in the late phase of type‐1 hypersensitivity reaction or in type‐2 and type‐3 delayed sensitivities is a matter that warrants further investigation.