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Simple Summary: Ochratoxin A (OTA), a nephrotoxic mycotoxin categorized as a possible carcinogenic agent, is a current problem in the cheese manufacturing industry. The main OTA producers are Penicillium nordicum and Penicillium verrucosum. Therefore, we aimed to investigate different cheese ripening phases to hinder OTA production. We elaborated cheese analogues, inoculated both fungal species, and performed two different ripening phases, at 10 and 15 °C. Despite the fact that both were able to grow, they did not express genes related to OTA production, and no OTA was detected during two weeks of ripening. Therefore, we show that, when the first phases of ripening are maintained at <15 °C, the possibility of OTA production is dramatically minimized. This information is of utmost importance for industries to enhance the safety of their cheese products, which positively impacts human health. A lack of control of the technological abiotic parameters apparent during cheese manufacture, including temperature and relative humidity, results in this dairy product being prone to mold contamination. Sometimes, inoculant molds are used to obtain the characteristic sensory properties of this type of product. However, during the maturation process, some unwanted molds can colonize the ripening cheese and produce mycotoxins. Mycotoxigenic molds such as Penicillium nordicum and Penicillium verrucosum can colonize ripened cheeses, contaminating them with ochratoxin A (OTA), a nephrotoxic 2B toxin. Thus, the presence of OTA in cheeses could represent a hazard to consumers' health. This study has evaluated the growth and OTA production of P. nordicum and P. verrucosum on a cheese analogue under simulated ripening conditions of 10 and 15 °C and 0.96 water activity (aw). Ecophysiological, molecular, and analytical tools assessed the mold growth, gene expression, and OTA production under these environmental conditions. Both species were able to effectively colonize the cheese under these ripening conditions. However, neither species expressed the otapks and otanps biosynthetic genes or produced phenotypic OTA. Therefore, these results suggest a relatively low risk of exposure to OTA for consumers of this type of cheese product. The conditions used were thus appropriate for cheese ripening to minimize the potential for contamination with such mycotoxins. An appropriate adjustment of the technological ripening parameters during such cheese manufacture could contribute to OTA-free cheeses. [ABSTRACT FROM AUTHOR]

Raspberry production is under threat from the emerging fungal pathogenic genus Cladosporium. We used amplicon‐sequencing, coupled with qPCR, to investigate how fruit age, fruit location within a polytunnel, polytunnel location and sampling date affected the fruit epiphytic microbiome. Fruit age was the most important factor impacting the fungal microbiome, followed by sampling date and polytunnel location. In contrast, polytunnel location and fruit age were important factors impacting the bacterial microbiome composition, followed by the sampling date. The within‐tunnel location had a small significant effect on the fungal microbiome and no effect on the bacterial microbiome. As fruit ripened, fungal diversity increased and the bacterial diversity decreased. Cladosporium was the most abundant fungus of the fruit epiphytic microbiome, accounting for nearly 44% of all fungal sequences. Rotorod air samplers were used to study how the concentration of airborne Cladosporium inoculum (quantified by qPCR) varied between location (inside and outside the polytunnel) and time (daytime vs. nighttime). Quantified Cladosporium DNA was significantly higher during the day than the night and inside the polytunnel than the outside. This study demonstrated the dynamic nature of epiphytic raspberry fruit microbiomes and airborne Cladosporium inoculum within polytunnels, which will impact disease risks on raspberry fruit. [ABSTRACT FROM AUTHOR]

There is little information on the effect of temperature, relative humidity (RH) and vapour pressure deficit (VPD) on the viable populations of two commercial biocontrol strains, Bacillus subtilis QST 713 (recently classified as B. velezensis) and Gliocladium catenulatum J1446 (syn. Clonostachys rosea). The PMAxx‐qPCR molecular assay was used to quantify the viable biocontrol agent (BCA) populations on fully extended lettuce and strawberry leaves under a range of temperature and RH combinations. Overall, there was a small decline in the population size of viable cells for the two biocontrol organisms on strawberry and lettuce leaves within 10 days of application. However, for most experimental runs, which contained general UK agronomy climates, such a decline was not statistically significant. Moreover, for a few runs, the viable populations increased significantly in optimal BCA growth temperatures with high RH. Only temperature (ambient) and dew point significantly affected the rate of temporal changes in the viable biocontrol population size. Thus, an increasing temperature led to decreased daily mortality. It should be noted that much of the variability in the estimated daily mortality rates remains unaccounted for, thus unless under extreme conditions, the biocontrol efficacy in practice is less likely to be affected by the survival of biocontrol microbes but more likely by other factors that influence the density of biocontrol cells in the phyllosphere, especially dilution due to rapid host leaf expansion, spray coverage and rain wash‐off. [ABSTRACT FROM AUTHOR]

Apple rootstock genotypes confer different levels of tolerance to apple replant disease (ARD) and vigour to a newly replanted apple tree. A hybrid management system of rotating the rootstock genotype planted between successive generations and inter-row planting in the alleyways of orchards may minimise the severity of ARD symptoms. High-throughput sequencing of the fungal ITS and bacterial 16S rDNA regions was used to investigate the diversity, and differential taxa present in soils displaying symptoms of ARD. Candidate pathogens and beneficial microorganisms were correlated with the above-ground establishment of each rootstock genotype in a UK cider orchard. Our results suggest that the same rootstock or rootstock with closely related parentage to the previous rootstock had more severe ARD symptoms. Planting in the alleyway appeared an effective strategy to minimise the severity of symptoms irrespective of rootstock genotype. The planting location effect had a higher contribution to the variation in the rhizosphere microbiome than that of the rootstock genotype. No predicted causal agents for ARD could be identified to a taxonomic level to predict their function but two species associated with mycorrhizae, Pteridiospora spinosispora and Paraglomus laccatum were identified as inversely correlated with ARD severity and could be candidate beneficial species for apple, warranting further investigation and research. Our findings suggest that planting in the alleyways and planting rootstocks genetically dissimilar to the previously planted rootstock can be beneficial for tree establishment. We have also identified species inversely associated with ARD severity, making candidates for future research to test the antagonistic effect of the species against ARD pathogens in apple roots. [ABSTRACT FROM AUTHOR]

Aim We assessed the effect of exposing apple orchard soil to different temperatures and CO2 levels on the resident microbiome of soils from a conventionally managed and an organically managed apple orchard. The key difference between these two orchards was that synthetic fertilizers and pesticides are routinely used in the former one. Methods and results To investigate the effect of CO2 and temperature, soil samples from each site at two depths were exposed to either elevated temperature (29°C) at either 5000 or 10 000 ppm for five weeks or control conditions (25°C + 400 ppm). Both bacterial and fungal communities were profiled with amplicon-sequencing. The differences between the two orchards were the most significant factor affecting the bacterial and fungal communities, contributing to 53.7–14.0% of the variance in Bray–Curtis β diversity, respectively. Elevated CO2 concentration and increased temperature affected organic orchard microbial diversity more than the conventionally managed orchard. A number of candidate beneficial and pathogenic microorganisms had differential abundances when temperature and CO2 were elevated, but their effect on the plant is unclear. Conclusions This study has highlighted that microbial communities in bulk soils are most significantly influenced by crop management practices compared to the climate conditions used in the study. The studied climate conditions had a more limited effect on microbial community diversity in conventionally managed soil samples than in organically managed soils. [ABSTRACT FROM AUTHOR]

Raspberry (Rosales: Rosaceae) production in the U.K. has moved rapidly in the last 10 years to under polythene, combined with a reduced availability of broad-spectrum fungicides. Hence, the incidence of previously less prevalent diseases, such as Cladosporium (Capnodiales: Cladosporiaceae), has largely increased. This study aimed to identify the predominant Cladosporium species on raspberry and to understand the nature of its infection on raspberry fruit. Raspberries were collected from farms across the U.K. and Spain and incubated; fungal isolates were then isolated from typical Cladosporium lesions and identified to the species level based on the sequences of the trans elongation factor α and actin genes. Cladosporium cladosporioides (Fres) de Vries was confirmed as the predominant species responsible for infecting raspberry fruit close to harvest on fruit from the U.K. and Spain, being present on 41.5% of U.K. fruit and 84.6% of Spanish fruit. Raspberries were subsequently inoculated at different developmental stages with C. cladosporioides isolates to determine the susceptibility to Cladosporium skin lesions and stigmata infections in relation to the developmental stage. Only the ripening and ripe raspberries were susceptible to Cladosporium, resulting in skin lesions. Cladosporium can colonise the stigmata of raspberries earlier in fruit development and future research is required to determine if such stigmata infections could cause subsequent skin lesion infections. This study has provided the necessary epidemiological information to develop effective management measures against the Cladosporium species. [ABSTRACT FROM AUTHOR]

Dry rot is an emerging issue for garlic production worldwide and Fusarium proliferatum is its major causal agent. Since the disease is seed-transmitted, sowing healthy cloves is crucial. In this study, some disinfection strategies were tested on garlic seeds, including steam, dry heat, chemical disinfectants and gaseous ozone (O3). Steam reduced the Colony Forming Units·g−1 (CFUs·g−1) by up to 92% in garlic seeds, but, at the same time, it affected their germination (−36%). Similarly, hydrogen peroxide (H2O2) and peracetic acid (C2H4O3) reduced the CFUs·g−1 by up to 83%; however, these methods also severely impaired germination (−40%). Dry heat did not negatively impact germination, but fungal contamination was not significantly reduced. The most promising strategy was gaseous O3 treatment; it decreased CFUs·g−1 by up to 96%, without causing any reduction of germination. The treatments applied were partially effective because the fungus is predominantly located in the outer layer of the seed, although it is also found in the inner portions. Some of these treatments can contribute to garlic protection from seed-borne pathogens and possibly reduce the occurrence of garlic dry rot. [ABSTRACT FROM AUTHOR]

Fusarium species can cause head blight of cereals worldwide. This is accompanied by impacts on yield and contamination of grains with mycotoxins. Regulations, with maximum limits, exist for the relevant Fusarium mycotoxins (e.g., type A and B trichothecenes, zearalenone and fumonisins). There is interest in a better understanding of the effect of key interacting abiotic factors which determine colonization and mycotoxin production in small grain cereals. Thus, this study examined the ecophysiological relationship between temperature and water availability (10–35 °C; water activity, aw, 0.87–0.98) on growth and production of Fusarium mycotoxins (zearelenone, ZEA; deoxynivalenol, DON; 3-acetyl deoxynivalenol, 3-Ac-DON and nivalenol, NIV) by three strains of F. asiaticum, a head blight pathogen isolated from China and becoming important in other global regions. These were carried out on simulated wheat-based matrices that identified the optimum (25 °C/0.98 aw) and marginal boundary conditions for growth (35 °C/0.90 aw) for all three strains. Contrarily, different mycotoxigenic profiles were observed between strains (p < 0.05). Four mycotoxins assessed were produced at 30 °C while cold temperature inhibited the production of NIV and ZEA, which were never detected at <20 and <15 °C, respectively. Optimal mycotoxin production conditions varied for each toxin with ZEA production which was best at 30 °C/0.93–0.95 aw, DON, 3-Ac-DON and NIV which was 0.98 aw/20–30 °C. Probabilistic models were used to predict growth and regulated mycotoxin production by the strains of F. asiaticum. This study will be beneficial in the development mitigation strategies for control of pre- and post-harvest colonization of cereals and mycotoxin contamination by this Fusarium species in cereals. [ABSTRACT FROM AUTHOR]

Pistachio nuts can become colonized by mycotoxigenic fungi, especially Aspergillus flavus, resulting in contamination with aflatoxins (AFs). We examined the effect of gaseous O3 (50–200 ppm; 30 min; 6 L/min) on (a) in vitro germination, (b) mycelial growth, and (c) aflatoxin B1 (AFB1) production on a milled pistachio nut-based medium at different water activity (aw) levels and at 30 °C. This was complimented with in situ studies exposing raw pistachio nuts to 50–200 ppm of O3. Exposure of conidia to gaseous O3 initially resulted in lower germination percentages at different aw levels. However, 12 h after treatment, conidial viability recovered with 100% germination after 24–48 h. Growth rates of mycelial colonies were slightly decreased with the increase of the O3 dose, with significant inhibition only at 0.98 aw. The production of AFB1 after O3 treatment and storage for 10 days was stimulated in A. flavus colonies at 0.98 aw. Raw pistachio nuts inoculated with A. flavus conidia prior to O3 exposure showed a significant decrease in population after 20 days of storage. However, AFB1 contamination was stimulated in most O3 treatments. The relationship between exposure concentration, time and prevailing aw levels on toxin control needs to be better understood for these nuts. [ABSTRACT FROM AUTHOR]

There is interest in understanding the relationship between naturally contaminated commodities and the potential for the production of different useful and toxic secondary metabolites (SMs). This study examined the impact of interacting abiotic stress parameters of water availability and temperature of stored naturally contaminated maize on the SM production profiles. Thus, the effect of steady-state storage water activity (aw; 0.80–0.95) and temperature (20–35 °C) conditions on SM production patterns in naturally contaminated maize was examined. The samples were analysed using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) to evaluate (a) the total number of known SMs, (b) their concentrations, and (c) changes under two-way interacting environmental stress conditions. A total of 151 metabolites were quantified. These included those produced by species of the Aspergillus, Fusarium and Penicillium genera and other unspecified ones by other fungi or bacteria. There were significant differences in the numbers of SMs produced under different sets of interacting environmental conditions. The highest total number of SMs (80+) were present in maize stored at 20–25 °C and 0.95 aw. In addition, there was a gradation of SM production with the least number of SMs (20–30) produced under the driest conditions of 0.80 aw at 20–30 °C. The only exception was at 35 °C, where different production patterns occurred. There were a total of 38 Aspergillus-related SMs, with most detected at >0.85 aw, regardless of the temperature in the 50–500 ng/g range. For Fusarium-related SMs, the pattern was different, with approx. 10–12 SMs detected under all aw × temperature conditions with >50% produced at 500 ng/g. A total of 40–45 Penicillium-related SMs (50–500 ng/g) were detected in the stored maize but predominantly at 20–25 °C and 0.95 aw. Fewer numbers of SMs were found under marginal interacting abiotic stress storage conditions in naturally contaminated maize. There were approx. eight other known fungal SM present, predominantly in low concentrations (<50 ng/g), regardless of interacting abiotic conditions. Other unspecified SMs present consisted of <20 in low concentrations. The effect of interacting abiotic stress factors for the production of different suites of SMs to take account of the different ecological niches of fungal genera may be beneficial for identifying biotechnologically useful SMs. [ABSTRACT FROM AUTHOR]

This study has examined the pattern of mycotoxin contamination of maize destined for animal feed in different global regions over a period of 3 years (2018–2020) with up to 1000+ samples analysed in each year. Overall, >75% of samples in each of the survey years were contaminated with multiple mycotoxins regardless of the global region (Europe, Africa, Asia, South Americas countries). Using LC-MS/MS, it was possible to quantify the relative contamination present in the samples in each year from the different regions of eight different mycotoxins including aflatoxin B1 (AFB1), ochratoxin A (OTA) deoxynivalenol (DON), fumonisin B1 (FB1) and B2, zearalenone (ZEA), T-2 and HT-2 toxins. The trends in mycotoxin contamination showed that there was a consistent contamination with DON in the 3 sampling years in all four regions. Interestingly, AFB1 contamination was prevalent in all regions in 2018, but more predominant in Europe and in 2019. In contrast, in 2020 it was found to be the major contaminant in Africa only. However, FB1 contamination of maize which was prevalent in Europe in 2018, became more prevalent in Asia and LATAM countries in 2019 and even in African maize in 2020. Comparisons of contamination with different mycotoxins in each of the years globally showed significant differences for AFB1, FB1, DON and ZEA between the different years. These results are discussed in relation to the trends of contamination of maize with mixtures of mycotoxins and the implication for their control in this key commodity used as an important ingredient in animal feed. [ABSTRACT FROM AUTHOR]

Background: Fusarium langsethiae is a T-2 and HT-2 mycotoxins producing species firstly characterised in 2004. It is commonly isolated from oats in Northern Europe. T-2 and HT-2 mycotoxins exhibit immunological and haemotological effects in animal health mainly through inhibition of protein, RNA and DNA synthesis. The development of a high-quality and comprehensively annotated assembly for this species is therefore essential in providing the molecular understanding and the mechanism of T-2 and HT-2 biosynthesis in F. langsethiae to help develop effective control strategies. Results: The F. langsethiae assembly was produced using PacBio long reads, which were then assembled independently using Canu, SMARTdenovo and Flye. A total of 19,336 coding genes were identified using RNA-Seq informed ab-initio gene prediction. Finally, predicting genes were annotated using the basic local alignment search tool (BLAST) against the NCBI non-redundant (NR) genome database and protein hits were annotated using InterProScan. Genes with blast hits were functionally annotated with Gene Ontology. Conclusions: We developed a high-quality genome assembly of a total length of 59 Mb and N50 of 3.51 Mb. Raw sequence reads and assembled genome is publicly available and can be downloaded from: GenBank under the accession JAFFKB000000000. All commands used to generate this assembly are accessible via GitHub: https://github.com/FadyMohareb/fusarium_langsethiae. [ABSTRACT FROM AUTHOR]

Hazelnuts are an economically important nut which is consumed world-wide. It is prone to infection by Aspergillus flavus and contamination with aflatoxins. Taking Tukey's hazelnuts as a research object, the objective of this study were to (a) quantify respiration rates and (b) dry matter losses (DMLs) and (c) aflatoxin B1 (AFB1) contamination of naturally stored shelled hazelnuts at different temperatures and water availabilities. Subsequently, shelled hazelnuts were inoculated with additional A. flavus inoculum prior to storage to examine effects on dry matter losses and on AFB1 contamination. Maximum respiration of hazelnuts and associated mycoflora was under wetter conditions of 0.90-0.95 water activity (aw = 12.5%-18% moisture content). This resulted in between approx. 10% DML at 25 and 30 after 5 days storage. Inoculation and storage of shelled hazelnuts + A. flavus inoculum resulted in similar patterns of respiration with optimal levels at 25-30 and >0.90 aw. Indeed, AFB1 contamination was highest at the maximum water levels tested of 0.90 aw. Indeed the contamination level exceeded the legislative limits set by the EU for AFB1 contamination of these nuts. Correlation between DMLs and all the AFB1 data in both sets of studies showed that very small changes in DML due to poor drying or storage of ≥0.6% resulted in AFB1 contamination levels exceeding the EU legislative limits. Thus, efficient drying and safe monitored storage is necessary to minimise the risk of AFB1 contamination in this economically important commodity and to avoid exposure of consumers to such toxins. [ABSTRACT FROM AUTHOR]

The objective of this study was to examine the effect of gaseous ozone (O3) on (a) germination, (b) mycelial growth, (c) aflatoxin B1 (AFB1) production by strains of Aspergillus flavus at different water activity (aw, 0.89-0.97=13.1%-24% moisture content) and 25 C in vitro. In addition, taking Egyptian peanuts as a research object, shelled peanuts at 0.93 aw were inoculated with two concentrations of A. flavus conidia (103, 105 conidia/g) and exposed to gaseous O3 and then stored. In all cases, exposure was for 30 min at 6 L/min. Generally, >100 ppm O3 significantly inhibited conidial germination of A. flavus strains (EGP-B07; SRRC-G 1907) on a defined yeast sucrose medium within 48 hrs. However, exposure of growing colonies of A. flavus to O3 with up to 300 ppm, had no effect on subsequent colony extension. The same concentration significantly affected AFB1 production, but only at 0.89 aw. Populations of both A. flavus strains in stored shelled peanuts exposed at 0.93 aw were significantly decreased at 100-400 ppm O3. However, AFB1 production was only significantly reduced in the 400 ppm O3 treatments at both inoculum load levels. These results are discussed in the context of O3 gas use for controlling A. flavus populations and AFB1 in shelled peanuts post-harvest. [ABSTRACT FROM AUTHOR]

Aflatoxins (AFs) are produced by fungi in crops and can cause liver cancer. Permitted levels are legislated and batches of grain are rejected based on average concentrations. Corn grown in Southern Georgia (GA), USA, which experiences drought during the mid-silk growth period in June, is particularly susceptible to infection by Aspergillus section Flavi species which produce AFs. Previous studies showed strong association between AFs and June weather. Risk factors were developed: June maximum temperatures > 33 °C and June rainfall < 50 mm, the 30-year normals for the region. Future climate data were estimated for each year (2000–2100) and county in southern GA using the RCP 4.5 and RCP 8.5 emissions scenarios. The number of counties with June maximum temperatures > 33 °C and rainfall < 50 mm increased and then plateaued for both emissions scenarios. The percentage of years thresholds were exceeded was greater for RCP 8.5 than RCP 4.5. The spatial distribution of high-risk counties changed over time. Results suggest corn growth distribution should be changed or adaptation strategies employed like planting resistant varieties, irrigating and planting earlier. There were significantly more counties exceeding thresholds in 2010–2040 compared to 2000–2030 suggesting that adaptation strategies should be employed as soon as possible. [ABSTRACT FROM AUTHOR]

Bread and intermediate moisture bakery products are mainly spoiled by yeasts and filamentous fungi. The inoculum load and preservation system used determines their shelf life. To extend the shelf life of such commodities, the use of chemical preservatives is the most common way to try and control the initiation of mold spoilage of bread. This study has utilized a rapid turbidimetric assay system (Bioscreen C) to examine the temporal efficacy of calcium propionate (CP) and potassium sorbate (PS) for controlling the growth of important bread spoilage fungi. The objectives were to compare the temporal growth of strains of three important spoilage fungi Hyphopichia burtonii (HB17), Paecilomyces variotii (PV11), and Penicillium roqueforti (PR06) isolated from visibly molded bread to (a) different concentrations of CP and PS (0–128 mM), (b) temperatures (25°C, 30°C), (c) water activity (aw; 0.95, 0.97), and (d) pH (5.0, 5.5). All three abiotic factors, pH, aw, and temperature, and preservative concentrations influenced the relative growth of the species examined. In general, PS was more effective than CP in inhibiting the growth of the strains of these three species. In addition, the Time to Detection (TTD) for the efficacy of the preservatives under the interacting abiotic factors was compared. The strain of Paecilomyces variotii (PV10) was the most tolerant to the preservatives, with the shortest TTD values for both preservatives. P. roqueforti was the most sensitive with the longest TTD values under all conditions examined. These results are discussed in the context of the evolution of resistance to food-grade preservatives by such spoilage fungi in bakery products. [ABSTRACT FROM AUTHOR]

The objective was to screen and evaluate the anti-fungal activity of lactic acid bacteria (LABs) isolated from Malaysian fermented foods against two Trichophyton species. A total of 66 LAB strains were screened using dual culture assays. This showed that four LAB strains were very effective in inhibiting growth of T. rubrum but not T. interdigitale. More detailed studies with Lactobacillus plantarum strain HT-W104-B1 showed that the supernatant was mainly responsible for inhibiting the growth of T. rubrum. The minimum inhibitory concentration (MIC), inhibitory concentration, the 50% growth inhibition (IC50) and minimum fungicide concentration (MFC) were 20 mg/mL, 14 mg/mL and 30 mg/mL, respectively. A total of six metabolites were found in the supernatant, with the two major metabolites being L-lactic acid (19.1 mg/g cell dry weight (CDW)) and acetic acid (2.2 mg/g CDW). A comparative study on keratin agar media showed that the natural mixture in the supernatants predominantly contained L-lactic and acetic acid, and this significantly controlled the growth of T. rubrum. The pure two individual compounds were less effective. Potential exists for application of the natural mixture of compounds for the treatment of skin infection by T. rubrum. [ABSTRACT FROM AUTHOR]

Temperatures approaching or exceeding 20°C have been measured during summer in polar regions at the surfaces of barren fellfield soils under cloudless skies around solar noon. However, despite the upper temperature limit for the growth of cold‐adapted microbes—which are abundant in polar soils and have pivotal roles in nutrient cycling—typically being close to this temperature, previous studies have not addressed the consequences of climate change for the metabolism of these organisms in the natural environment. Here in a 5‐year field experiment on Alexander Island in the southern maritime Antarctic, we show that the abundance of Pseudogymnoascus roseus, the most widespread decomposer fungus in maritime Antarctic fellfield soils, is reduced by 1–2 orders of magnitude when irrigated and nutrient‐amended soils are warmed to >20°C during summer. Laboratory experiments under conditions mimicking those during midsummer in the natural environment indicated that the hyphal extension rates of P. roseus isolates and the activities of five extracellular enzymes are reduced by 54%–96% at high water availability after exposure to temperatures cycling daily from 2 to 21°C and 2 to 24°C, relative to temperatures cycling from 2 to 18°C. Given that the temperatures of surface soils at the study site already reach 19°C during midsummer, the observations reported here suggest that, at predicted rates of warming arising from moderate greenhouse gas emissions, inhibitory effects of climate change on the metabolism of P. roseus could manifest themselves within the next few decades. Furthermore, with peak temperatures at the surfaces of fellfield soils at other maritime Antarctic locations and in High Arctic and alpine regions already exceeding 20°C during summer, the observations suggest that climate warming has the potential to inhibit the growth of other cold‐adapted microbes, with negative effects on soils as the Earth's climate continues to warm. [ABSTRACT FROM AUTHOR]

Pistachio nuts are an economically important commodity produced by many countries. They can be colonized by mycotoxigenic fungi, especially Aspergillus flavus , resulting in contamination with aflatoxins (AFs) , especially aflatoxin B1 (AFB1), a Class 1a carcinogen. The objectives were to examine the effect of interactions between the two key abiotic factors, temperature and water activity (a w ) on (a) in vitro growth and AFB1 production by four strains of A. flavus isolated from pistachio nuts, on a milled pistachio nut medium modified ionically (NaCl) and non-ionically (glycerol) in the range 20–35°C and 0.995–0.85 a w , (b) colonization of layers of raw pistachio nuts stored at different interacting temperature x a w conditions and on relative AFB1 production and (c) develop models to produce contour maps of the optimal and marginal boundary conditions for growth and AFB1 production by up to 4 strains of this species. On pistachio nut-based media, optimum growth of four strains of A. flavus was at 0.98–0.95 a w and 30–35°C. Optimum AFB1 production was at 30–35°C and 0.98 a w . No significant differences in growth was found on ionic and non-ionically modified media. Colonization of layers of raw pistachio nuts was slower and contamination with AFB1 significantly less than in in vitro studies. Contour maps based on the pooled data for up to four strains (in vitro , in situ) showed the optimum and marginal conditions for growth and AFB1 production. These data can be used to identify those conditions which represent a high, intermediate or low risk of colonization and AFB1 contamination in the pistachio nut processing chain. These results are discussed in the context of the development of appropriate intervention strategies to minimize AFB1 contamination of this economically important commodity. [ABSTRACT FROM AUTHOR]

We report a previously undescribed member of the Helotiales that is superabundant in soils at two maritime Antarctic islands under Antarctic Hairgrass (Deschampsia antarctica Desv.). High throughput sequencing showed that up to 92% of DNA reads, and 68% of RNA reads, in soils from the islands were accounted for by the fungus. Sequencing of the large subunit region of ribosomal (r)DNA places the fungus close to the Pezizellaceae, Porodiplodiaceae, and Sclerotiniaceae, with analyses of internal transcribed spacer regions of r DNA indicating that it has affinities to previously unnamed soil and root fungi from alpine, cool temperate and Low Arctic regions. The fungus was found to be most frequent in soils containing C aged to 1,000–1,200 years before present. The relative abundances of its DNA and RNA reads were positively associated with soil carbon and nitrogen concentrations and δ13C values, with the relative abundance of its DNA being negatively associated with soil pH value. An isolate of the fungus produces flask-shaped phialides with a pronounced venter bearing masses of conidia measuring 4.5–6(7) × 1.8–2.5 μm, suggestive of anamorphic Chalara. Enzymatic studies indicate that the isolate strongly synthesizes the extracellular enzyme acid phosphatase, and also exhibits alkaline phosphatase and naphthol-AS-BI-phosphohydrolase activities. Ecophysiological measurements indicate optimal hyphal growth of the isolate at a pH of 4.2–4.5 and a water potential of −0.66 MPa. The isolate is a psychrotroph, exhibiting measureable hyphal growth at −2°C, optimal hyphal extension rate at 15°C and negligible growth at 25°C. It is proposed that the rising temperatures that are predicted to occur in maritime Antarctica later this century will increase the growth rate of the fungus, with the potential loss of ancient C from soils. Analyses using the GlobalFungi Database indicate that the fungus is present in cold, acidic soils on all continents. We advocate further studies to identify whether it is superabundant in soils under D. antarctica elsewhere in maritime Antarctica, and for further isolates to be obtained so that the species can be formally described. [ABSTRACT FROM AUTHOR]

During grape cultivation and wine production, the most effective way to prevent ochratoxin A (OTA) contamination of grapes and wine is to control ochratoxigenic fungal species, especially Aspergillus carbonarius, using appropriate cultivation techniques. In this study, the influence of an organic farming system (OFS) and an integrated farming system (IFS) on the incidence of A. carbonarius on grapes, and OTA contamination of wine, were examined. Mycological analysis of grapes collected from Kotsifali cultivar (Vitis vinifera L.) vineyards and grown under two farming systems (OFS and IFS) was performed over two growing seasons. For the same two growing seasons, OTA levels of representative wine samples from wineries located in the same area, made from the same cultivar (single varietal or covinificated with Mandilari), and grown under the two farming systems were determined. The results showed that the farming system had a significant influence on the incidence of A. carbonarius, with IFS being the most effective in the control of the fungus and the prevention of OTA occurrence in wine. This knowledge could offer viticulturists a useful tool to produce safer grapes, giving winemakers an incentive to make low-OTA wine. [ABSTRACT FROM AUTHOR]

Botrytis cinerea, Mucor piriformis and Rhizopus spp. are mainly responsible for post-harvest fruit rot in strawberry in the UK. However, research to date has been focused on the epidemiology and management of B. cinerea in strawberry (i.e., grey mould). A number of experiments were carried out to study the epidemiology of M. piriformis and Rhizopus on strawberry, including flower/fruit susceptibility, inoculum dose, in vitro spore germination and infection of both detached and attached fruit. The results showed that M. piriformis and Rhizopus had very similar epidemiological characteristics. Both pathogens were unable to infect flowers, but fruit became increasingly susceptible from the green development stage onwards. In vitro germination required near-saturation humidity conditions and was reduced in both low and high temperatures. However, infection of attached ripe fruit and subsequent rot development were relatively unaffected by temperature and relative humidity conditions that commonly occur under field conditions in the UK. On detached ripe fruit, a high spore infection potency was observed for both pathogens. Infection of fruit was completed within 9 to 24 h. Thus, the most important factor determining the level of infection of fruit by M. piriformis and Rhizopus in UK commercial crops is probably the number of spores deposited on the fruit. Management strategies should be targeted at reducing inoculum concentrations within crops in order to minimise the risk of fruit infection by M. piriformis and Rhizopus. [ABSTRACT FROM AUTHOR]

In this review, we present the current information on development and applications of biological control against phytopathogenic organisms as well as mycotoxigenic fungi in Malaysia as part of the integrated pest management (IPM) programs in a collective effort to achieve food security. Although the biological control of phytopathogenic organisms of economically important crops is well established and widely practiced in Malaysia with considerable success, the same cannot be said for mycotoxigenic fungi. This is surprising because the year round hot and humid Malaysian tropical climate is very conducive for the colonization of mycotoxigenic fungi and the potential contamination with mycotoxins. This suggests that less focus has been made on the control of mycotoxigenic species in the genera Aspergillus, Fusarium, and Penicillium in Malaysia, despite the food security and health implications of exposure to the mycotoxins produced by these species. At present, there is limited research in Malaysia related to biological control of the key mycotoxins, especially aflatoxins, Fusarium‐related mycotoxins, and ochratoxin A, in key food and feed chains. The expected threats of climate change, its impacts on both plant physiology and the proliferation of mycotoxigenic fungi, and the contamination of food and feed commodities with mycotoxins, including the discovery of masked mycotoxins, will pose significant new global challenges that will impact on mycotoxin management strategies in food and feed crops worldwide. Future research, especially in Malaysia, should urgently focus on these challenges to develop IPM strategies that include biological control for minimizing mycotoxins in economically important food and feed chains for the benefit of ensuring food safety and food security under climate change scenarios. [ABSTRACT FROM AUTHOR]

There has been significant interest in the development of formulations of non-toxigenic strains of Aspergillus flavus for control of toxigenic strains to reduce the aflatoxin B1 (AFB1) contamination of maize. In the future, climate change (CC) abiotic conditions of temperature (+2–4°C), CO2 (existing levels of 400 vs. 800–1,200 ppb), and drought stress will impact on the agronomy and control of pests and diseases. This study has examined (1) the effect of two-way interacting factors of water activity × temperature on colonization and AFB1 contamination of maize cobs of different ripening ages; (2) the effect of non-toxigenic strains of A. flavus (50:50 inoculum ratio) on relative control of toxigenic A. flavus and AFB1 contamination of ripening cobs; (3) post-harvest control of AFB1 by non-toxigenic strains of A. flavus in non-GM and isogenic GM maize cultivars using the same inoculum ratio; and (4) the impact of three-way interacting CC factors on relative control of AFB1 in maize cobs pre-harvest and in stored non-GM/GM cultivars. Pre-harvest colonization and AFB1 production by a toxigenic A. flavus strain was conserved at 37°C when compared with 30°C, at the three ripening stages of cob development examined: milk ripe (R3), dough (R4), and dent (R5). However, pre-harvest biocontrol with a non-toxigenic strain was only effective at the R3 and R4 stages and not at the R5 stage. This was supported by relative expression of the aflR regulatory biosynthetic gene in the different treatments. When exposed to three-way interacting CC factors for control of AFB1 pre-harvest, the non-toxigenic A. flavus strain was effective at R3 and £4 stages but not at the R5 stage. Post-harvest storage of non-GM and GM cultivars showed that control was achievable at 30°C, with slightly better control in GM-cultivars in terms of the overall inhibition of AFB1 production. However, in stored maize, the non-toxigenic strains of A. flavus had conserved biocontrol of AFB1 contamination, especially in the GM-maize cultivars under three-way interacting CC conditions (37°C × 1,000 ppm CO2 and drought stress). This was supported by the relative expression of the aflR gene in these treatments. This study suggests that the choice of the biocontrol strains, for pre- or post-harvest control, needs to take into account their resilience in CC-related abiotic conditions to ensure that control of AFB1 contamination can be conserved. [ABSTRACT FROM AUTHOR]

The capacity of five white rot fungi species to degrade linuron, metribuzin and chlorpyrifos when applied both as single pesticides and mixed together in different concentrations on nutritionally poor media was investigated. Our results suggested that Pleurotus ostreatus, Pycnoporus coccineus, Phlebiopsis gigatea and Τrametes versicolor showed a remarkable tolerance to the pesticides, in all media tested. The EC50 values presented a noticeable difference in the mixtures as compared with the individual ones. The minimum growth rate in the mixture was obtained by P. ostreatus whereas P. coccineus appeared to be more efficient than the rest fungal isolates, when cultivated in the soil extract medium. P. coccineus, P. gigantea and T. versicolor produce high levels of polyphenol oxidase, but only T. versicolor was capable of decomposing linuron when combined with metribuzin and chlorpyrifos. [ABSTRACT FROM AUTHOR]

Changes in environmental stress impact on secondary metabolite (SM) production profiles. Few studies have examined targeted SM production patterns in relation to interacting environmental conditions in stored cereals. The objectives were to examine the effect of water activity (aw; 0.95-0.90) x temperature (10-25 °C) on SM production on naturally contaminated stored wheat and that inoculated with Fusarium graminearum. Samples were analysed using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) on (a) total number of known SMs, (b) their concentrations and (c) changes under environmental stress. 24 Fusarium metabolites were quantified. Interestingly, statistical differences (ChisSq., p < 0.001) were observed in the number of SMs produced under different sets of interacting environmental conditions. The dominant metabolites in natural stored grain were deoxynivalenol (DON) and nivalenol (NIV) followed by a range of enniatins (A, A1, B, B1), apicidin and DON-3-glucoside at 10 °C. Increasing temperature promoted the biosynthesis of other SMs such as aurofusarin, moniliformin, zearalenone (ZEN) and their derivatives. Natural wheat + F. graminearum inoculation resulted in a significant increase in the number of metabolites produced (ChisSq., p < 0.001). For ZEN and its derivatives, more was produced under cooler storage conditions. Fusarin C was enhanced in contrast to that for the enniatin group. The relative ratios of certain groups of targeted SM changed with environmental stress. Both temperature and aw affected the amounts of metabolites present, especially of DON and ZEN. This study suggests that the dominant SMs produced in stored temperate cereals are the mycotoxins for which legislation exists. However, there are changes in the ratios of key metabolites which could influence the relative contamination with individual compounds. Thus, in the future, under more extreme environmental stresses, different dominant SMs may be formed which could make present legislation out of step with the future contamination which might occur. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) contamination from Fusarium graminearum colonization is particularly important in food and feed wheat, especially during post-harvest storage with legislative limits for both food and feed grain. Indicators of the relative risk from exceeding these limits would be useful. We examined the effect of different water activities (aw; 0.95-0.90) and temperature (10-25 °C) in naturally contaminated and irradiated wheat grain, both inoculated with F. graminearum and stored for 15 days on (a) respiration rate; (b) dry matter losses (DML); (c) ZEN production and (d) relationship between DML and ZEN contamination relative to the EU legislative limits. Gas Chromatography was used to measure the temporal respiration rates and the total accumulated CO2 production. There was an increase in temporal CO2 production rates in wetter and warmer conditions in all treatments, with the highest respiration in the 25 °C × 0.95 aw treatments + F. graminearum inoculation. This was reflected in the total accumulated CO2 in the treatments. The maximum DMLs were in the 0.95 aw/20-25 °C treatments and at 10 °C/0.95 aw. The DMLs were modelled to produce contour maps of the environmental conditions resulting in maximum/minimum losses. Contamination with ZEN/ZEN-related compounds were quantified. Maximum production was at 25 °C/0.95-0.93 aw and 20 °C/0.95 aw. ZEN contamination levels plotted against DMLs for all the treatments showed that at ca. <1.0% DML, there was a low risk of ZEN contamination exceeding EU legislative limits, while at >1.0% DML, the risk was high. This type of data is important in building a database for the development of a post-harvest decision support system for relative risks of different mycotoxins. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is a saprophytic fungus that may colonize several important crops, including cotton, maize, peanuts and tree nuts. Concomitant with A. flavus colonization is its potential to secrete mycotoxins, of which the most prominent is aflatoxin. Temperature, water activity (aw) and carbon dioxide (CO2) are three environmental factors shown to influence the fungus-plant interaction, which are predicted to undergo significant changes in the next century. In this study, we used RNA sequencing to better understand the transcriptomic response of the fungus to aw, temperature, and elevated CO2 levels. We demonstrate that aflatoxin (AFB1) production on maize grain was altered by water availability, temperature and CO2. RNA-Sequencing data indicated that several genes, and in particular those involved in the biosynthesis of secondary metabolites, exhibit different responses to water availability or temperature stress depending on the atmospheric CO2 content. Other gene categories affected by CO2 levels alone (350 ppm vs. 1000 ppm at 30 °C/0.99 aw), included amino acid metabolism and folate biosynthesis. Finally, we identified two gene networks significantly influenced by changes in CO2 levels that contain several genes related to cellular replication and transcription. These results demonstrate that changes in atmospheric CO2 under climate change scenarios greatly influences the response of A. flavus to water and temperature when colonizing maize grain. [ABSTRACT FROM AUTHOR]

Heavy metal–contaminated soils are a serious environmental problem. Herein, the culturable heterotrophic bacterial community present on two metal(loid)-contaminated sites in the Northern Portugal was investigated. The bacterial counts ranged from 5.96 to 7.69 and 7.04 to 7.51 (log CFU g−1soil) in Sites 1 and 3, respectively. The bacterial population was predominantly composed ofFirmicutes, Proteobacteria, andActinobacteriaon both sites. The most represented genera in Site 1 wereBacillus(41%) andPseudomonas(27%), whereasArthrobacter(21%) andPseudomonas(13%) were the most represented genera in Site 3. Several bacterial isolates showed tolerance to high concentrations of metal(loid)s, suggesting that both contaminated sites are a valuable source of metal(loid)-tolerant bacteria, which may be further used in bioremediation and/or phytoremediation processes. [ABSTRACT FROM PUBLISHER]

For the most-extreme fungal xerophiles, metabolic activity and cell division typically halts between 0.700 and 0.640 water activity (approximately 70.0-64.0% relative humidity). Here, we investigate whether glycerol can enhance xerophile germination under acute water-activity regimes, using an experimental system which represents the biophysical limit of Earth's biosphere. Spores from a variety of species, including Aspergillus penicillioides, Eurotium halophilicum, Xerochrysium xerophilum (formerly Chrysosporium xerophilum) and Xeromyces bisporus, were produced by cultures growing on media supplemented with glycerol (and contained up to 189 mg glycerol g dry spores−1). The ability of these spores to germinate, and the kinetics of germination, were then determined on a range of media designed to recreate stresses experienced in microbial habitats or anthropogenic systems (with water-activities from 0.765 to 0.575). For A. penicillioides, Eurotium amstelodami, E. halophilicum, X. xerophilum and X. bisporus, germination occurred at lower water-activities than previously recorded (0.640, 0.685, 0.651, 0.664 and 0.637 respectively). In addition, the kinetics of germination at low water-activities were substantially faster than those reported previously. Extrapolations indicated theoretical water-activity minima below these values; as low as 0.570 for A. penicillioides and X. bisporus. Glycerol is present at high concentrations (up to molar levels) in many types of microbial habitat. We discuss the likely role of glycerol in expanding the water-activity limit for microbial cell function in relation to temporal constraints and location of the microbial cell or habitat. The findings reported here have also critical implications for understanding the extremes of Earth's biosphere; for understanding the potency of disease-causing microorganisms; and in biotechnologies that operate at the limits of microbial function. [ABSTRACT FROM AUTHOR]

This study aimed at the isolation and characterization of metal(loid)-tolerant bacteria from the rhizosphere of Phragmites australis and Juncus effusus plants growing in two long-term contaminated sites in Northern Portugal. Site 1 had higher contamination than Site 3. Bacteria were isolated using metal(loid)-supplemented (Cd, Zn, and As) media. Isolates were grouped by random amplified polymorphic DNA and identified by 16S rRNA gene sequencing. Strains were also examined for their metal(loid) tolerance. The counts of metal(loid)-tolerant bacteria were higher in Site 1 and ranged between log 7.17 CFU g−1 soil in As-containing medium and log 7.57 CFU g−1 soil in Zn-containing medium, while counts at Site 3 varied between log 5.33 CFU g−1 soil in Cd-containing medium and log 6.97 CFU g−1 soil in As-containing medium. The composition of bacterial populations varied between locations. In Site 1, the classes Actinobacteria (36%) and Bacilli (24%) were well represented, while in Site 3 strains were mainly affiliated to classes Actinobacteria (35%), γ- Proteobacteria (35%), and β- Proteobacteria (12%). The order of metal(loid) toxicity for the isolated strains was Cd > As > Zn. Overall, 10 strains grew at 500 mg Cd L−1, 1000 mg Zn L−1, and 500 mg As L−1, being considered the most metal(loid)-tolerant bacteria. These strains belonged to genera Cupriavidus, Burkholderia, Novosphingobium, Sphingobacterium, Castellaniella, Mesorhizobium, Chryseobacterium, and Rhodococcus and were mainly retrieved from Site 1. The multiple metal(loid)-tolerant strains isolated in this study have potential to be used in bioremediation/phytoremediation. [ABSTRACT FROM AUTHOR]

This paper examines the impact that single and interacting environmental stress factors have on tolerance mechanisms, molecular ecology and the relationship with secondary metabolite production by a group of mycotoxigenic species of economic importance. Growth of these fungi ( Aspergillus flavus, A.ochraceus, A.carbonarius, Penicillium nordicum and P. verrucosum) is influenced by water and temperature interactions and type of solute used to induce water stress. Such abiotic stresses are overcome by the synthesis of increased amounts of low molecular weight sugar alcohols, especially glycerol and erythritol, to enable them to remain active under abiotic stress. This is accompanied by increased expression of sugar transporter genes, e.g., in A. flavus, which provides the nutritional means of tolerating such stress. The optimum conditions of water activity ( a) × temperature stress for growth are often different from those for secondary metabolite production. The genes for toxin production are clustered together and their relative expression is influenced by abiotic interacting stress factors. For example., A. flavus synthesises aflatoxins under water stress in non-ionic solutes. In contrast, P. nordicum specifically occupies a high salt (0.87 a = 22 % NaCl) niche such as cured meats, and produces ochratoxin A (OTA). There is differential and temporal expression of the genes in the secondary metabolite clusters in response to a × temperature stress. We have used a microarray and integrated data on growth, relative expression of key genes in the biosynthetic pathways for secondary metabolite production and toxin production using a mixed growth model. This was used to correlate these factors and predict the toxin levels produced under different abiotic stress conditions. This system approach to integrate these different data sets and model the relationships could be a powerful tool for predicting the relative toxin production under extreme stress conditions, including climate change scenarios. This approach will facilitate a better functional understanding of the influence that environmental stress has on these mycotoxigenic fungi and enable better prevention strategies to be developed based on this system-based approach. [ABSTRACT FROM AUTHOR]

Whereas osmotic stress response induced by solutes has been well-characterized in fungi, less is known about the other activities of environmentally ubiquitous substances. The latest methodologies to define, identify and quantify chaotropicity, i.e. substance-induced destabilization of macromolecular systems, now enable new insights into microbial stress biology (Cray et al. in Curr Opin Biotechnol 33:228-259, , doi:; Ball and Hallsworth in Phys Chem Chem Phys 17:8297-8305, , doi:; Cray et al. in Environ Microbiol 15:287-296, , doi:). We used Aspergillus wentii, a paradigm for extreme solute-tolerant fungal xerophiles, alongside yeast cell and enzyme models ( Saccharomyces cerevisiae and glucose-6-phosphate dehydrogenase) and an agar-gelation assay, to determine growth-rate inhibition, intracellular compatible solutes, cell turgor, inhibition of enzyme activity, substrate water activity, and stressor chaotropicity for 12 chemically diverse solutes. These stressors were found to be: (i) osmotically active (and typically macromolecule-stabilizing kosmotropes), including NaCl and sorbitol; (ii) weakly to moderately chaotropic and non-osmotic, these were ethanol, urea, ethylene glycol; (iii) highly chaotropic and osmotically active, i.e. NHNO, MgCl, guanidine hydrochloride, and CaCl; or (iv) inhibitory due primarily to low water activity, i.e. glycerol. At ≤0.974 water activity, Aspergillus cultured on osmotically active stressors accumulated low- M polyols to ≥100 mg g dry weight. Lower- M polyols (i.e. glycerol, erythritol and arabitol) were shown to be more effective for osmotic adjustment; for higher- M polyols such as mannitol, and the disaccharide trehalose, water-activity values for saturated solutions are too high to be effective; i.e. 0.978 and 0.970 (25 ºC). The highly chaotropic, osmotically active substances exhibited a stressful level of chaotropicity at physiologically relevant concentrations (20.0-85.7 kJ kg). We hypothesized that the kosmotropicity of compatible solutes can neutralize chaotropicity, and tested this via in-vitro agar-gelation assays for the model chaotropes urea, NHNO, phenol and MgCl. Of the kosmotropic compatible solutes, the most-effective protectants were trimethylamine oxide and betaine; but proline, dimethyl sulfoxide, sorbitol, and trehalose were also effective, depending on the chaotrope. Glycerol, by contrast (a chaotropic compatible solute used as a negative control) was relatively ineffective. The kosmotropic activity of compatible solutes is discussed as one mechanism by which these substances can mitigate the activities of chaotropic stressors in vivo. Collectively, these data demonstrate that some substances concomitantly induce chaotropicity-mediated and osmotic stresses, and that compatible solutes ultimately define the biotic window for fungal growth and metabolism. The findings have implications for the validity of ecophysiological classifications such as 'halophile' and 'polyextremophile'; potential contamination of life-support systems used for space exploration; and control of mycotoxigenic fungi in the food-supply chain. [ABSTRACT FROM AUTHOR]

Alternaria species are common saprophytes or pathogens of a wide range of plants pre- and post-harvest. This review considers the relative importance of Alternaria species, their ecology, competitiveness, production of mycotoxins and the prevalence of the predominant mycotoxins in different food products. The available toxicity data on these toxins and the potential future impacts of Alternaria species and their toxicity in food products pre- and post-harvest are discussed. The growth of Alternaria species is influenced by interacting abiotic factors, especially water activity (aw), temperature and pH. The boundary conditions which allow growth and toxin production have been identified in relation to different matrices including cereal grain, sorghum, cottonseed, tomato, and soya beans. The competitiveness of Alternaria species is related to their water stress tolerance, hydrolytic enzyme production and ability to produce mycotoxins. The relationship between A. tenuissima and other phyllosphere fungi has been examined and the relative competitiveness determined using both an Index of Dominance (ID) and the Niche Overlap Index (NOI) based on carbon-utilisation patterns. The toxicology of some of the Alternaria mycotoxins have been studied; however, some data are still lacking. The isolation of Alternaria toxins in different food products including processed products is reviewed. The future implications of Alternaria colonization/infection and the role of their mycotoxins in food production chains pre- and post-harvest are discussed. [ABSTRACT FROM AUTHOR]

This investigated the roles of water activity (aw) and fungicides on the competitiveness of two Fusarium verticillioides strains against other spoilage fungi commonly present in maize (F. proliferatum, Aspergillus niger, A. flavus, A. ochraceus and Penicillium verrucosum). Fungal strains were inoculated on artificial media containing maize flour. The effects were determined of three aw levels (0.99, 0.98 and 0.95) and three fungicides (tebuconazole, procloraz and prothioconazole) on fungal interactions, the Index of Dominance (ID) of isolates and fumonisin B1+B2 (FBs) production. The two strains of F. verticillioides showed similar behaviour in conditions where water was freely available (0.99 aw); at 0.98 and 0.95 aw both F. verticilliodes strains had the lowest total ID scores (8-6 and 10-12, respectively). They showed the same ability to compete against other fungi having the highest ID scores against P. verrucosum and A. ochraceus and the lowest against A. niger and A. flavus. The lowest water activity gave (0.95 aw) was the most conducive for fumonisin production with significant differences to 0.98 and 0.99 aw. In a co-inoculation experiment, only FBs production from P. verrucosum was greater in the presence of the F. verticilliodes strains other fungi. The use of fungicides reduced Indices of Dominancy (ID) for both F. verticilliodes strains. A significant reduction in F. verticilloides growth was observed when combining water stress and fungicide treatments. This information provides increased understanding of the colonisation patterns of F. verticillioides in relation to other mycobiota and to both environmental and chemical stresses, and has implications in relation to future climate change scenarios. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effects of calcium propionate, water activity (aw) and incubation time on the total fungal count and aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) production in the broiler finisher feed. The feed was added with calcium propionate (5 g kg–1), adjusted to 0.85, 0.90 and 0.95 aw and stored for 28 days at 25°C, analysing for mould growth and aflatoxins production every 7 days. Analysis of variance indicated that all the factors (preservative, aw and storage time) alone and in combination significantly (p < 0.001) affected the total fungal count and aflatoxins production in the feed. Minimum total fungal counts (1.99 × 102 CFU g–1) were observed in calcium propionate feed at 0.85 aw on day 1 and the highest (4.36 × 109 CFUs g–1) in control sample at 0.95 aw on day 28 of storage. During the storage period, AFB1 content in control samples increased from 11.35 to 73.44, from 11.58 to 81.81 and from 11.54 to 102.68 ng g–1, whereas in preserved feed the content of B1 increased from 11.47 to 37.83, from 11.54 to 49.07 and from 11.20 to 53.14 ng g–1 at 0.85, 0.90 and 0.95 aw, respectively. Similar patterns were noted for AFB2, AFG1 and AFG2 contents. All the aflatoxins readily increased over storage time; however, the increase was much slower in preserved feed that contained a lower amount of available water. This study reveals that calcium propionate addition to poultry litter along with water activity amelioration is an effective tool for controlling mould incidence and aflatoxin production in poultry feed. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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 x 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 x temperature x elevated CO2 (2x and 3x 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. [ABSTRACT FROM AUTHOR]

Phlomis purpurea (Lamiaceae), found in Quercus suber and Quercus ilex ssp. rotundifolia forest habitats in southern Portugal, is a non-host for the oomycete Phytophthora cinnamomi, the main biotic factor involved in cork oak and holm oak decline in the Iberian Peninsula. The effect of P. purpurea crude ethanol root extract was evaluated in vitro on P. cinnamomi mycelial growth, sporangial production, zoospore release and germination as well as on chlamydospore production and viability. The protection of cork oak against infection by the pathogen was also evaluated in planta. At 10 mg ml, in vitro inhibition of the pathogen structures was 85-100 %. In addition, P. purpurea plants were shown to protect Q. suber and Q. ilex from P. cinnamomi infection and to reduce the inoculum potential in glasshouse trials, indicating the ability to reduce root infection by the pathogen. The results suggest that P. purpurea has the potential to reduce disease spread and that their root extracts could provide candidate substances for control of the important pathogen, P. cinnamomi. [ABSTRACT FROM AUTHOR]

Grey mould rot (Botrytis cinerea) development in vitro or in eggplant (Solanum melongena L.) fruit was evaluated after treatment with dittany (Origanum dictamnus L.) oil (DIT) and storage at 12°C and 95% relative humidity during or following exposure to the volatiles. DIT volatiles used in different concentration (0-50-100-250 μL/L) and times of exposure (up to 120 h) examined the effects on pathogen development as well as fruit quality parameters. In vitro, fungal colony growth was inhibited with the application of DIT oil (during or after exposure) and/or time of application. Continuous exposure to oils reduced conidial germination and production with fungistatic effects observed in 250 μL/L. In vivo, fungal lesion growth and conidial production reduced in DITtreated fruits. Interesting, in fruits preexposed to volatiles before fungal inoculation, DIT application induced fruit resistance against the pathogen, by reduced lesion growth and conidial production. Conidial viability reduced in >100 μL/L DIT oil. Fruits exposed to essential oil did not affect fruit quality related attributes in general, while skin lightness (L value) increased in 50 and 100 μL/L DIT oil. The results of the current study indicated that dittany volatiles may be considered as an alternative food preservative, eliminating disease spread in the storage/transit atmospheres. [ABSTRACT FROM AUTHOR]

Samples of stored maize from villages located in five different agroecological zones (southern lowlands, northern lowlands, Senqu river valley, foothills and mountains) of Lesotho were collected in 2009/10 and 2010/11 and assessed for contamination with toxigenic fungi. The water activity of all samples collected during the two seasons was <0.70. The total fungal populations of the maize from different regions in the two seasons was not significantly different ( p > 0.05). Fusarium verticillioides, F. proliferatum and F. subglutinans predominated in different regions in both seasons based on molecular analyses. In the 2009/10 season, the isolates of these species all produced FB, while in the 2010/11 season, very few produced FB. A. flavus isolates (2009/10) were recovered from mountains and Senqu river valley samples while the 2010/11 isolates were predominantly from the foothills and northern lowlands. The mountain isolates of Aspergillus section Flavi produced the highest levels of AFB (20 mg kg). Aspergillus parasiticus was only isolated from the foothills, Senqu river valley and southern lowlands samples, and the AFB levels produced ranged from 'none detected' to 3.5 mg kg. The Aspergillus ochraceous isolates were least frequently encountered in both seasons. In the 2009/10 season, the isolates from the northern lowlands produced ochratoxin A (OTA) in culture. No isolates of A. niger from different regions in both seasons produced any OTA. Multi-mycotoxin analyses of the maize samples were done for a range of mycotoxins. At least one sample from each region in both seasons was FB-positive. FB levels for 2010/11 samples (7-936 μg kg) were higher than in the 2009/10 season (2-3 μg kg). In both seasons, the mountains registered the highest levels of FB. Deoxynivalenol (DON) was recovered from all the samples analysed, with the highest mean contamination of 1,469 μg kg in samples from the northern lowlands. Moniliformin (MON) was detected from all agroecological zones in the two seasons (5-320 μg kg in 2009/10; 15-1,205 μg kg in 2010/11). Emerging toxins such as fusaproliferin (FUS) and beauvericin (BEA) were also detected. OTA was not detected in any of the samples analysed. Only one 2009/10 sample in the Senqu river valley was positive for AFB. This is the first report on toxigenic fungi and multi-mycotoxin contamination of maize samples from subsistence farmers' stores in different agroecological zones of Lesotho. [ABSTRACT FROM AUTHOR]

The potential of three atoxigenic strains from different geographical origins in Africa were examined forin vitroandin situcompetitiveness against two toxigenic strains ofAspergillus flavusand subsequent inhibition of aflatoxin B1(AFB1) production under different environmental conditions. Temperature, water activity (aw) and substrate influenced the types of interaction between the three AFL−and two AFL+strains. The competitiveness and AFB1reduction ability of the three atoxigenic strains when interacting with the two toxigenic strains were evaluated by inoculation of 100, 25:75, 50:50, 75:25 and 100% ratios of mixed spore suspensionsin vitroon malt extract and milled maize agars over 28 days andin situon stored maize grain for 14 days, respectively at 0.99, 0.96 and 0.90aw. For all the treatments, the effect ofawand inoculum ratio and their interaction was highly significant. Toxin inhibition was >80%in vitroat both 0.99 and 0.96aw.In situAFB1reduction was influenced by the toxigenic strain assayed,awand the inoculum ratio. Where control was achieved, it was more variable at 0.96aw, while with more stringent water stress conditions (0.90aw) the percentage inhibition was up to 77.2%. The study shows the importance of including environmental factors in screening and identifying effective atoxigenic strains for control of AFs (aflatoxins). [ABSTRACT FROM AUTHOR]

To study the effects of and interactions among feed types, seasons, and agroecological zones on the total fungal viable count and aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), and G2 (AFG2) production in poultry feed, an experiment was conducted using three-factorial design. A total of 216 samples of poultry feed ingredients, viz. maize, wheat, rice, cotton seed meal (CSM), and finished products, that is, starter and finisher broilers' rations, were collected from Peshawar, Swat, and D. I. Khan districts of Khyber Pakhtunkhwa, Pakistan, during the winter, spring, summer, and autumn seasons of the year 2007/2008. Analysis of variance showed that there was a complex interaction among all these factors and that this influenced the total fungal viable count and relative concentrations of the aflatoxins produced. Minimum total culturable fungi (6.43 × 10 CFUs/g) were counted in CSM from D. I. Khan region in winter season while maximum (26.68 × 10 CFUs/g) in starter ration from Peshawar region in summer. Maximum concentrations of AFB1 (191.65 ng/g), AFB2 (86.85 ng/g), and AFG2 (89.90 ng/g) were examined during the summer season whereas the concentration of AFG1 was maximum (167.82 ng/g) in autumn in finisher ration from Peshawar region. Minimum aflatoxins were produced in the winter season across all the three agroecological zones. [ABSTRACT FROM AUTHOR]

This study examined the relationship between storage environmental factors (water activity (aw) (0.89–0.97) and temperature (15°C–30°C)), colonisation of wheat and maize by Fusarium graminearum and F. verticillioides respectively and the dry matter losses (DMLs) caused and quantified by contamination with deoxynivalenol (DON), zearalenone (ZEA) and fumonisins (FUMs) during storage. Fungal growth was assessed by the amount of CO2 produced under different interacting conditions of aw and temperature. DMLs were quantified using the cumulative CO2 data, and these were shown to increase as temperature and aw increased. The amount of DON, ZEA (wheat for human consumption) and FUMs (feed maize) produced was significantly affected by the storage conditions. The three toxins however showed different patterns of production. Optimum for DON was at the wettest conditions (0.97aw) and the highest temperature assessed (30°C), whereas for ZEA this shifted to 25°C. FUMs were produced in higher amounts in maize at 30°C and 0.97aw; however, at intermediate aw levels (0.955aw), the highest production occurred at 25°C followed by 20°C. Polynomial models were developed for the effect of the storage factors on DMLs and toxin production. DMLs under different environmental conditions were significantly correlated with DON and FUMs. DON contamination was above the EU limits in at least 80% of the wheat samples with DMLs >1%, whereas at least 70% of the same samples contained ZEA above the respective EU legislative limits. Similarly, at least 75% of the maize samples with DMLs ≥ 0.9% exceeded the EU limits for the sum of FUMs in feed. These results show that it may be possible to use temporal CO2 production during storage of grains as an indicator of the level of contamination of the grain with mycotoxins. [ABSTRACT FROM AUTHOR]

The aims of this study were to use qualitative volatile fingerprints obtained using a hybrid sensor array system to screen anti-fungals for controlling the important lung infecting fungus, Aspergillus fumigatus, especially in immunocompromised patients. SIFT-MS was also used to try and identify key volatiles produced by A. fumigatus. Initial studies were carried out to identify the ED and ED (effective dose) for inhibiting growth of A. fumigatus using three anti-fungal compounds, benomyl, tebuconazole and fluconazole. Subsequent studies involved inoculation of malt extract agar plates with spores of A. fumigatus (25 and 37°C) over periods of 24-72 h to examine the headspace volatile fingerprints generated from the sample treatments using the hybrid sensor array system to compare controls and ED/ED concentrations. The sensor responses showed discrimination between treatments after 48-h incubation when benomyl and tebuconazole were used against A. fumigatus at 37°C using Principal Components Analysis and Cluster Analysis. SIFT-MS analysis showed that methyl pentadiene, ethanol, isoprene and methanol were key biomarker volatiles produced by A. fumigatus in the presence of anti-fungal compounds. This may also be a good approach for the development of rapid screening of anti-microbial compounds and potentially useful for monitoring the possible build up of resistance to specific drug types. Volatile fingerprints produced by patient samples could also be used to evaluate whether lung infections are caused by bacteria or specific fungi to facilitate early diagnosis and enable the right drug treatment to be prescribed. [ABSTRACT FROM AUTHOR]

The temporal efficacy of three different chemical fungicides (Folicur®, Proline®, Sportak 45EW®) and a biocontrol bacterium (Serenade, B. subtilis) in reducing growth and toxin production by isolates of F. verticillioides and A. flavus was studied in vitro under different water activity regimes (0.99, 0.98 and 0.95). All the fungicides significantly inhibited mycelial growth compared with the control; the most effective treatment, both against F. verticillioides and A. flavus, was Sportak 45EW® (approx. 99%). The inhibitory effect of all fungicides generally improved with increasing concentration. Serenade always decreased fungal growth, with optimal results at concentrations of 104 and 106 (70-75% reduction). All the fungicide treatments resulted in a significant reduction in both FB1+FB2 and AFB1 production when compared to the control, at the end of the incubation period and with the 2 concentrations used (approx. 99%). A threshold concentration inoculum of at least 104 CFUs of B. subtilis per g was required to achieve a significant control of mycotoxin production. Sportak 45EW® and Serenade gave the best control of mycotoxin production with a reduction of 95% compared to the controls. Use of Serenade in the field should include due consideration to its sensitivity to low water activities, when compared to the target pathogens. [ABSTRACT FROM AUTHOR]