Mycotoxigenic fungi in peanuts from different geographic regions of Egypt

To understand the importance of mycotoxigenic fungi in Egyptian peanuts, samples from five regions (Alexandria, El-Beheira, El-Sharqiya, El-Daqahelaya in northern Egypt and Asyut, southern Egypt) in two seasons (2007, 2008) were collected. Aspergillus was consistently the most frequent genus in seeds and in-shell peanuts and was the dominant mycotoxigenic component of the mycobiota. There was no direct correlation between the moisture content of the samples and the fungal populations on peanut seeds tested from different regions. The most common species were from Aspergillus section Flavi (4.7-78.3%), Aspergillus section Nigri (9.4–52.6%) and Aspergillus section Circumdati (5.1–30.9%). In the in-shell peanut samples, the lowest populations were recorded in El-Beheira and Asyut (3.7–4.0 log₁₀ CFU g^-1) and the highest in Alexandria and Elsharqiya (4.1–6.0 log₁₀ CFU g^-1). Aspergillus section Flavi and section Nigri were the most dominant, and Aspergillus section Circumdati were only found in samples in 2008. Both qualitative (coconut cream agar) and quantitative analyses (HPLC) were used to analyse the potential mycotoxin production by strains isolated from peanuts. Of a total of 88 Aspergillus section Flavi strains examined, 95% were A. flavus based on production of aflatoxin B₁ on yeast extract sucrose (YES) medium and confirmation using molecular analyses. Of 64 Aspergillus section Circumdati strains only 28% produced ochratoxin A (OTA), and were identified as A. westerdijkiae. No Aspergillus section Nigri strains produced OTA, and they were identified as A. niger (uniseriate). The presence of these toxigenic fungi indicates that there is a potential risk of mycotoxin contamination in Egyptian peanuts and suggests that problems can arise from contamination with both aflatoxins and perhaps also OTA.     

Toxigenic fungi associated with stored corn

A total of 246 fungal isolates were obtained from 25 moldy corn samples collected in central Iowa. Either water or ether extracts of all corn samples except one exhibited some degree of toxicity in mice and/or ducklings. At least one toxigenic isolate was obtained from each corn sample. Extracts of cultures of 99 of the fungal isolates, involving 13 genera, produced death in one or more of the assay animals. The majority of the toxigenic isolates belonged to the generaAspergillus andPenicillium. Three isolates ofTrichothecium roseum were highly toxic to ducklings and mice. The ducklings exhibited a flaccid paralysis shortly after receiving an oral dose of extracts of these three isolates. Death frequently occurred following a second oral dose given 24 hr later. Mice given intraperitoneal injections of extracts ofT. roseum first exhibited lethargy and intermittent tremors, then hyperemotivity, roughening of the hair coat, abdominal respirations, incoordination, dyspnea and clonic convulsions with death occurring usually within 10 to 20 min.

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Zusammenfassung Insgesamt sind 246 Pilzisolierungen von 25 verschimmelten Getreideproben, die in Zentral-Iowa gesammelt worden sind, erhalten worden. Sowohol Wasser- wie Aether-Extrakte von allen Getreideproben zeigten eine gewisse Toxizität in Mäusen als auch in Entchen außer einer einzigen Probe. Kulturextrakte von 99 der Pilzisolierungen, die 13 Gattungen umfaßten, verursachten den Tod in einem oder in mehreren Versuchstieren. Die Mehrheit der toxischen Isolierungen gehörte den GattungenAspergillus undPenicillium an. Drei Isolierungen vonTrichothecium roseum waren hoch toxisch für Entchen und Mäuse. Die Entchen zeigten eine schlaffe Paralyse kurz nachdem sie eine orale Dosis der Auszüge dieser drei Isolierungen erhalten hatten. Tod erfolgte häufig nach der zweiten oralen Dosis innerhalb 24 Stunden. Mäuse, die eine intraperitoneale Injektion der Auszüge vonT. roseum erhielten, zeigten erst eine Lethargie und einen intermittierenden Tremor, dann eine Hyperemotivität, aufgerauhte Haarbedeckung, abdominale Atmung, Inkoordination, Dyspnoe, klonische Zuckungen und Tod gewöhnlich innerhalb 10 bis 20 Minuten.

     

Mycotoxigenic fungi contaminating wheat; toxicity of different Alternaria compacta strains

We studied mycotoxigenic fungi contaminating stored wheat grain, measured the toxins they secreted, and assessed their harmfulness. We focused on one common genus Alternaria, and chose 19 isolates representing A. compacta to study how different strains differed in their mycotoxin secretion and toxicity. Toxicity was assessed in a bioassay with a model bacteria Bacillus subtilis. All 19 A. compacta strains secreted toxins. Both the mycotoxin pattern and the fungal toxicity differed between the A. compacta stains. It seemed that some other toxins than alternariols or altenue acted as the main virulence factors of A. compacta against B. subtilis. We suggest that the most commonly studied mycotoxins do not necessarily indicate the toxicity of the fungi. The high variation in the amounts and toxins that different Alternaria species and strains secrete pose a challenge to the food supply chain.     

Mycotoxigenic potential of fungi isolated from freshly harvested Argentinean blueberries

Alternaria alternata, A. tenuissima, Fusarium graminearum, F. semitectum, F. verticillioides, Aspergillus flavus, and Aspergillus section Nigri strains obtained from blueberries during the 2009 and 2010 harvest season from Entre Ríos, Argentina were analyzed to determine their mycotoxigenic potential. Taxonomy status at the specific level was determined both on morphological and molecular grounds. Alternariol (AOH), alternariol monomethyl ether (AME), aflatoxins (AFs), zearalenone (ZEA), fumonisins (FBs), and ochratoxin A (OTA) were analyzed by HPLC and the trichotecenes deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin (HT-2), T-2 toxin (T-2), fusarenone X (FUS-X), 3-acetyl-deoxynivalenol (3-AcDON), and 15-acetyl-deoxynivalenol (15-AcDON) by GC. Twenty-five out of forty two strains were able to produce some of the mycotoxins analyzed. Fifteen strains of Aspergillus section Nigri were capable of producing Fumonisin B₁ (FB₁); two of them also produced Fumonisin B₂ (FB₂) and one Fumonisin B₃ (FB₃). One of the F. graminearum isolated produced ZEA, HT-2, and T-2 and the other one was capable of producing ZEA and DON. Two A. alternata isolates produced AOH and AME. Four A. tenuissima were capable of producing AOH and three of them produced AME as well. One Aspergillu flavus strain produced aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂), and aflatoxin G₁ (AFG₁). To our knowledge, this is the first report showing mycotoxigenic capacity of fungal species isolated from blueberries that include other fungi than Alternaria spp.     

Ecology of fungi associated with moist stored barley grain

A study of the ecology of fungi associated with moist stored barley grain was carried out on samples from a partially sealed silo in Cumberland. Samples were collected during harvest and after storage at high moisture content in a partially sealed silo. At harvest, the field fungi Alternaria, Fusarium, Cladosporium and Mucor spp. were dominant both in the husks and in the dehusked, surface-sterilized grain but they were rapidly succeeded by the storage fungi: Aspergillus, Penicillium and Absidia spp. then constituted about 80 % of the initial invasion. At this stage internal grain infection rarely exceeded 5 % compared to 65–80 % infection in the husks. In subsequent months, there was a remarkable increase in invasion, mainly by Penicillium, Aspergillus and Absidia spp. and by yeasts, which appeared both in the husks and in dehusked grain. In the third month of storage, Penicillium cyclopium, P. roqueforti, Absidia corymbifera, Aspergillus candidus, A. terreus and yeasts became dominant. The thermophilous species Aspergillus fumigatus, A. terreus, Absidia corymbifera, Mucorpusillus and Dactylomyces crustaceus were isolated from husks and in dehusked grain incubated at 37 and 45 °C. Most grains were infected significantly by single genera.     

Chromosomal regions associated with segregation distortion in maize

Segregation distortion skews the genotypic frequencies from their Mendelian expectations. Our objectives in this study were to assess the frequency of occurrence of segregation distortion in maize, identify chromosomal regions consistently associated with segregation distortion, and examine the effects of gametophytic factors on linkage mapping. We constructed a simple sequence repeat (SSR) linkage map for a LH200/LH216 F₂Syn3 (i.e., random-mated three times) population, and compared the segregation distortion in this map with the segregation distortion in three published linkage maps. Among 1,820 codominant markers across the four mapping populations, 301 (17%) showed segregation distortion (P < 0.05). The frequency of markers showing segregation distortion ranged from 19% in the Tx303/CO159 mapping population to 36% in the B73/Mo17 mapping population. A positive relationship was found between the number of meioses and the frequency of segregation distortion detected in a population. On a given chromosome, nearly all of the markers showing segregation distortion favored the allele from the same parent. A total of 18 chromosomal regions on the ten maize chromosomes were associated with segregation distortion. The consistent location of these chromosomal regions in four populations suggested the presence of segregation distortion regions (SDRs). Three known gametophytic factors are possible genetic causes of these SDRs. As shown in previous research, segregation distortion does not affect the estimate of map distance when only one gametophytic factor is present in an SDR.     

Effect of chlorine dioxide gas on fungi and mycotoxins associated with sick building syndrome

The growth of indoor molds and their resulting products (e.g., spores and mycotoxins) can present health hazards for human beings. The efficacy of chlorine dioxide gas as a fumigation treatment for inactivating sick building syndrome-related fungi and their mycotoxins was evaluated. Filter papers (15 per organism) featuring growth of Stachybotrys chartarum, Chaetomium globosum, Penicillium chrysogenum, and Cladosporium cladosporioides were placed in gas chambers containing chlorine dioxide gas at either 500 or 1,000 ppm for 24 h. C. globosum was exposed to the gas both as colonies and as ascospores without asci and perithecia. After treatment, all organisms were tested for colony growth using an agar plating technique. Colonies of S. chartarum were also tested for toxicity using a yeast toxicity assay with a high specificity for trichothecene mycotoxins. Results showed that chlorine dioxide gas at both concentrations completely inactivated all organisms except for C. globosum colonies which were inactivated an average of 89%. More than 99% of ascospores of C. globosum were nonculturable. For all ascospore counts, mean test readings were lower than the controls (P < 0.001), indicating that some ascospores may also have been destroyed. Colonies of S. chartarum were still toxic after treatment. These data show that chlorine dioxide gas can be effective to a degree as a fumigant for the inactivation of certain fungal colonies, that the perithecia of C. globosum can play a slightly protective role for the ascospores and that S. chartarum, while affected by the fumigation treatment, still remains toxic.     

Diversity and functioning of fungi associated with leaf litter decomposition in Asian forests of different climatic regions

The pattern of diversity and functioning of fungi associated with leaf litter decomposition in Asian forests of different climatic regions was investigated by performing meta-analysis of published data for seven tree species in subalpine, temperate, subtropical and tropical forests. Fungal assemblages were examined by using common standard isolation-culture methods, and the abilities of individual fungal species to decompose leaf litter were examined with pure culture decomposition tests. The climatic patterns of diversity, assemblage structure and genus composition depended on the method of isolation: the washing method revealed no consistent pattern, whereas the surface sterilization method showed lower diversity and greater relative abundance of dominant fungal species within the assemblages in cooler climates. The decomposing ability of species within fungal assemblages was greater in warmer than in cooler climates and in broad-leaved than in coniferous tree species. In particular, the greatest abilities to cause mass loss were found among fungi with ligninolytic activity in broad-leaved tree species in warmer climates.     

Molecular genetic analysis of maize mitochondrial regions associated with CMS

The Mmolecular-genetic polymorphism of 86 maize lines of world and Ukrainian breeding with S-, C- and T-types of cytoplasmic male sterility (CMS) and with wild-type mitochondra was studied by PCR analysis of mitochondrion regions. A molecular marker system was able to detect and identify a certain type of CMS in the maize lines and differentiate maize lines with a certain type of CMS from both lines with a different type of CMS and those with the wild-type cytoplasm.     

OTA-producing fungi isolated from stored cocoa beans

Aims:  The aim of this study was to identify fungal populations in unroasted cocoa beans stored in Spain in order to evaluate the ochratoxin A (OTA)-production ability of certain Aspergillus isolates.
Methods and Results:  Twenty batches of cocoa beans from different origins and with different OTA content were selected for this study. Three Aspergillus carbonarius and 13 Aspergillus niger aggregate strains isolated from these cocoa bean samples were selected to evaluate their OTA synthesis ability, being the only A. carbonarius isolates which are OTA producers [−1 culture medium; LOD = 6  μ g kg⁻¹ culture medium].
Conclusions:  No correspondence was found between the OTA levels in cocoa beans and the presence of OTA-producing fungi. Nonetheless, some samples contained A. carbonarius with a high OTA-producing ability and, consequently, specific fungal controls should be set up during storage to avoid this toxin.
Significance and Impact of the Study:  Toxigenic fungi in cocoa beans are not well understood. This study attempted to identify these fungi and evaluate their OTA-producing ability.     

Fusarium species complex and mycotoxins in grain maize from maize hybrid trials and from grower's fields

Aims: To quantify and to compare the occurrence of Fusarium species in maize kernels and stalk pieces, to analyse mycotoxins in kernels and maize crop residues, to evaluate two approaches to obtain kernel samples and to compare two methods for mycotoxin analyses. Methods and Results: The occurrence of Fusarium species in maize kernels and stalk pieces from a three‐year maize hybrid trial and 12 kernel samples from grower’s fields was assessed. Nine to 16 different Fusarium species were detected in maize kernels and stalks. In kernels, F. graminearum, F. verticillioides and F. proliferatum were the most prevalent species whereas in stalks, they were F. equiseti, F. proliferatum and F. verticillioides. In 2006, 68% of the kernel samples exceeded the recommended limit for pig feed for deoxynivalenol (DON) and 42% for zearalenone (ZON), respectively. Similarly, 75% of the samples from grower’s fields exceeded the limits for DON and 50% for ZON. In maize crop residues, toxin concentrations ranged from 2·6 to 15·3 mg kg^?1 for DON and from 0·7 to 7·4 mg kg^?1 for ZON. Both approaches to obtain maize kernel samples were valid, and a strong correlation between mycotoxin analysis using ELISA and LC‐MS/MS was found. Conclusions: The contamination of maize kernels, stalk pieces and remaining crop residues with various mycotoxins could pose a risk not only to animal health but also to the environment. With the hand‐picked sample, the entire Fusarium complex can be estimated, whereas combine harvested samples are more representative for the mycotoxin contents in harvested goods. Significance and Impact of the Study: This is the first multi‐year study investigating mycotoxin contamination in maize kernels as well as in crop residues. The results indicate a high need to identify cropping factors influencing the infection of maize by Fusarium species to establish recommendations for growers.     

Molecular-genetic analysis of maize mitochondrion regions associated with cytoplasmic male sterility

Molecular-genetic polymorphism of 86 world and Ukrainian breeding maize lines with S-, C- and T-types of cytoplasmic male sterility (CMS) and with normal wild type mitochondrion has been researched via mitochondrion regions PCR-analysis. Molecular marker system allowed to detect and identify definite type of CMS within maize lines, as well as to differentiate lines with definite CMS type either from lines with another CMS type or from normal wild type cytoplasm lines.     

Ergosterol and Fusarium mycotoxins content in two maize cultivars under different forms of nitrogen fertilizers

Mycotoxins are secondary metabolites produced by many filamentous fungi, such as Fusarium, and are widespread in nature and can adversely affect almost all organisms. Mycotoxins can be formed in very different conditions such as when plants are growing or in crops that are stored in improper conditions. One of the methods to prevent the formation of mycotoxins in powdered feed is using varieties of appropriate quality and proper mineral fertilization, primarily nitrogen. In this study, the concentration of ergosterol and mycotoxins in the kernels of two maize cultivars from different maturity groups in Poland during the harvest of 2011 and 2012 was investigated using HPLC methods. We aimed to analyse the concentration of ergosterol as a fungal biomass indicator and mycotoxins from two maize cultivars under six different forms of nitrogen fertilizers. The “stay‐green” hybrid ES Paroli was characterized by a significantly lower ergosterol and the ability to accumulate the mycotoxins, compared to the classic cultivar ES Palazzo. The varieties of maize that remain green for a longer time period, the so‐called stay‐green type, are characterized by a different coefficient of nitrogen remobilization compared to the classic hybrids. Their dynamics of growth and accumulation of nutrients indicates a nitrogen fertilization system, indicating that slow‐release fertilizers are potentially more adapted to the cycles of maize vegetation. Hence, compared to the classic variant, the “stay‐green” variety uses nitrogen more effectively from mineral fertilizers. Such hybrids are healthier and remain in good condition for a longer time; therefore, they are characterized by a lower pathogen pressure. Thus, the cultivation of such varieties can be considered as one of the elements of integrated maize production. We observed identical reactions of the studied experimental factors for both vegetation seasons, which indicates the lack of environmental impact on the functioning and interaction of experimental factors.     

Toxicity of Fusarium mycotoxins on maize plants

Synergistic effects of Fusarium — toxins mixture (moniliformin, fumonisin B₁, fusaproliferin, zearalenone, zearalenol and deoxynivalenol, each at concentration 3.5 μg mL-1) on maize plants of resistant and susceptible cultivars were studied. After 72-hour treatment the biomass production with both cultivars was approximately 6% lower than in the respective controls. In the resistant cultivar chlorophyll content was increased comparing to control, with higher increase in chlorophyll b. In susceptible cultivar chlorophyll content was slightly decreased, particularly with chlorophyll b.No significant differences between treated and non-treated plants were found in the cell ultrastructure. In susceptible plants the young root cells were more vacuolated and plasmolysis occurred in the cells of outer cortex. In leaves of this cultivar also disorganization of thylakoids in some chloroplasts was observed.     

Different populations of bacteria associated with sheathed and bare regions of roots of field-grown maize

It has been previously shown that soil sheaths cling tightly to some portions of all axile roots and cover all but the growing tips of the young roots of field-grown maize. These sheaths overlie immature regions of the roots which have intact epidermal cells with root hairs, and living, thus non-conducting, late metaxylem elements. Loss of the soil sheath in the proximal region coincides with the opening of these large metaxylem vessels. Now, total, and viable counts have been recorded of bacteria associated with the root surface and adhering soil of sheathed and bare regions. These showed some common features, in that populations of similar size were associated with the two root regions in plants beginning to flower. Each population included about the same numbers of bacteria that were viable on each of three selective media (nitrogen-free, Pseudomonas F or MacConkey). However, more spore-formers capable of growth on nitrogen-free media and more fluorescent bacteria were isolated from the sheathed regions. Actinomycetes were absent from sheathed but plentiful on bare regions.The high numbers of diverse types of bacteria associated with both root surfaces can be related to the previously demonstrated similarity in amounts of organic carbon released from each region. The proliferation of actionomycetes on the bare roots and their exclusion from sheathed roots may in part be due to the lower water status of the bare region, which is related to its greater axial conducting capacity. Thus the distribution of the two types of root surface within an individual root system has important implications for the choice of root and rhizosphere sampling techniques and for root bacterization work.     

Transpiration inhibition by stored xylem sap from well-watered maize plants

There is increasing evidence that a chemical signal exists in xylem sap of plants subjected to water deficits which influences physiological responses in plant shoots. An important method of studying this signal is the transpiration response of excised leaves exposed to xylem sap collected from plants. However, Munns et al [Plant, Cell & Environment 16, 867–877] cautioned that transpiration inhibition is observed when xylem sap collected from wheat and barley is stored before determining physiological activity. The objective of the study reported here was to determine if transpiration inhibition develops in maize sap collected from well-watered plants when the sap is stored under various conditions. It was found that storage of maize sap collected from well-watered plants for only 1 d at -20°C resulted in the development of substantial transpiration inhibition in bioassay leaves. Storage of sap at 4°C resulted in the development of the effect after 2 weeks, while storage at ?86°C showed only small transpiration inhibition after 3 weeks. The major source of the transpiration inhibition was the development of a substance in the stored sap that resulted in physical blockage of the transpiration stream in bioassay leaves. However, a small signal component may also have developed in the stored sap. Because of the possibility of ionic activity under freezing conditions at ?20°C, calcium was studied for its potential involvement in the transpiration inhibition. However, the calcium concentrations found to inhibit transpiration were nearly an order of magnitude larger than the calcium concentrations observed in xylem sap.     

A survey on the occurrence of mycotoxins in wheat and maize from western Romania

Samples of wheat (n = 25) and maize (n = 30) for animal consumption, collected in 1997 after harvest from western Romania, were analyzed by enzyme immunoassays for mycotoxin contamination. Toxins analyses included deoxynivalenol (DON), 3-acetylDON, 15- acetylDON, fusarenone X (FX), T-2 Toxin (T-2), diacetoxyscirpenol (DAS), zearalenone (ZEA), fumonisin B1 (FB1), aflatoxin B1 (AFB1), ochratoxin A (OA), and citrinin (CT). DON and acetylDONs were the major contaminants in wheat (100%) and maize (46%). Median values for DON, 3-acetylDON, and 15-acetylDON were 880 μg kg-1, 66 μg kg- 1, and 150 μg kg-1 in wheat, and 890 μg kg-1, 180 μg kg-1, and 620 μg kg- 1 in maize, respectively. Additionally, 3,15-diacetylDON was detected in some samples by HPLC-EIA analysis. All samples were negative for FX (<150 μg kg-1). T-2 was found in wheat (n = 6) and maize (n = 1) at levels between 13 and 63 μg kg- 1. DAS (2.6 μg kg-1) was found in one maize sample. ZEA occurred in all wheat and in four maize samples, median values were 10 μg kg-1 and 250 μg kg-1, respectively. One maize sample contained FB1 (140 μg kg-1). All samples were AFB1-negative (<4 μg kg-1). OA was found in one wheat sample (37 μg kg- 1), CT was found in one maize sample (580 μg kg- 1). This first reported natural occurrence of a range of mycotoxins in Romanian feeding stuff shows that DON and acetyl DONs may be present at levels which may affect animal production. This revised version was published online in June 2006 with corrections to the Cover Date.     

Production of trichothecene and non-trichothecene mycotoxins by Fusarium species isolated from maize in Minnesota

Eighty-two cultures of Fusarium species isolated in 1986 from moldy maize in Minnesota were each cultured on rice for 4 weeks and found to produce the following mycotoxins: F. graminearum isolates, deoxynivalenol (DON, 4–225 g/g), 3-acetyldeoxynivalenol (3-ADON, 2–4g/g), 15-acetyldeoxynivalenol (15-ADON, 1–35 g/g) and zearalenone (ZEA, 5–4350 g/g); F. moniliforme, fusarin C (detectable amounts to 1000 g/g); F. mòniliforme, F. oxysporum, F. proliferatum and F. subglutinans isolates, moniliformin (15–6775 g/g); F. moniliforme, F. proliferatum, and F. subglutinans isolates, fusaric acid (detectable amounts). Other mycotoxins screened for in each rice sample and not detected were T-2 toxin, HT-2 toxin, neosolaniol, T-2 tetraol, nivalenol, fusarenon-X, scirpenols, alpha and beta trans-zearalenols, wortmannin, and fusarochromanone. The rat feeding bioassay indicated that other, unidentified toxins may be present.