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Powdery mildew caused by Podosphaera cerasi is the most important fungal disease of sweet cherries in the Pacific Northwest of the United States. In this study, several factors related to disease epidemiology were evaluated. The experiments were conducted to investigate flower susceptibility to P. cerasi infection by in planta and in vitro inoculation. The susceptibility of fruit at various developmental stages was investigated using defined concentrations of P. cerasi conidia. Furthermore, the threshold of conidial concentration required for fruit infection was determined. The pathogen activity during full bloom was limited and not related to fruit disease incidence and severity at harvest. Foliar infections always preceded fruit infections by an average of 42 days during the 3 years of the study. The onset of fruit infection followed, on average, 66 days after full bloom and appeared simultaneously on all susceptible cherry cultivars in the research orchard. Disease symptoms were only observed on fruit in Biologische Bundesanstalt, Bundessortenamt, and Chemical Industry scale 8 (maturity) in all cultivars examined. During this stage, a concentration of 500 conidia/ml was sufficient to cause fruit infection at harvest. Interaction between the inoculation dates and conidial concentration revealed a dependency of disease development on the host stage at the time of inoculation; the younger the fruit, the more conidia are needed to cause disease at harvest. Molecular studies showed a rapid increase in conidia viability at the transition from asymptomatic to the symptomatic disease of fruit. No evidence of ontogenic resistance of fruit to powdery mildew infection was observed.

is the main aflatoxin producer in food and feed and has wide ecological niches. Contamination of food products such as pistachio nuts and aflatoxin secretion directly affects food safety and intern...

Powdery mildew caused by

In Iran, Phytophthora crown and root rot of pistachio trees (also known as gummosis) destroys a significant number of fertile and non-fertile trees each year. To identify potential biocontrol agents effective in controlling pistachio gummosis, 200 soil samples were collected from 19 pistachio growing regions in Kerman province. Out of the 321 strains tested for antagonistic activity against Phytophthora pistaciae, 13 were selected as potential inhibitors of the disease. The tested strains were able to inhibit Phytophthora growth in dual culture, volatile and non-volatile assays by 14–72%, 12–68% and 27–85%, respectively. The highest inhibition was achieved by three strains identified as Bacillus subtilis using phenotypic characteristics, biochemical and physiological tests as well as sequencing the 16S rRNA genomic region for each strain. Co-inoculation experiments of six months old pistachio seedlings with P. pistaciae and the three selected B. subtilis strains reduced mortality rates by up to 80%. B. subtilis strain BSIPR35 was identified as the most promising biocontrol agent in greenhouse experiments. The ability of the selected strains to withstand drought, high temperature and salinity stresses was further tested. The growth of the strains was reduced under abiotic stresses ranging from 22 to 94%. All strains had the same growth under drought stress, while in salinity and under high temperature strain BSIPR35 acted superiorly compared to the other two strains. The bacterial strains may be effective in inhibiting gummosis in vivo, which requires further investigations.

A large-scale study was conducted to assess which of the five most accessible hermetic storage devices on the Kenyan market fulfill the needs of smallholder farmers by positively impacting three major areas of concern: insect infestation, grain quality, and mycotoxin (aflatoxin and fumonisin) contamination. Efficacy of two hermetic silos (plastic and metal) and three hermetic bags (PICS, GrainPro's GrainSafe™, and Super Grain) was directly compared to current maize storage in polypropylene (PP) bags under local environmental conditions using representative storage volumes during a 6-month storage period. Impact of maize grain stored at typical (∼15%) and recommended (

Resistant cultivars of hop (Humulus lupulus) have been grown, with the aim of helping to manage powdery mildew in the Pacific Northwest since the first report of the disease in the field in 1997 (4). A major objective of many breeding programs is development of resistance to powdery mildew, and this has generally been achieved by single resistance genes (qualitative resistance). One such gene, R6 (3), has been utilized extensively in new cultivars and has prevented epidemics of the disease in those cultivars across the Pacific Northwestern United States for approximately 15 years. In 2011, a grower in Washington State reported outbreaks of powdery mildew on cv. Apollo, which is thought to possess powdery mildew resistance derived from R6. Fungicides and cultural control measures were applied, and the grower reported no substantial crop damage from the disease. During the winter of 2012, the same grower planted rhizomes of cv. Apollo in a greenhouse in the Yakima Valley of Washington State and later found the plants to be affected by powdery mildew. Affected leaves from plants of cvs. Apollo, Newport, and Nugget (all reported [3] or assumed to possess R6 based on pedigree) grown in the same greenhouse were later provided to the authors. Conidia obtained from each affected plants were transferred to plants of the highly susceptible cv. Symphony, which is not known to contain any resistance genes. After 10 to 14 days of incubation, resultant conidia from each cultivar above (total of three isolates) were transferred to greenhouse grown plants of cvs. Nugget and Symphony and incubated at 18°C. Within 7 days, all three isolates produced powdery mildew colonies characteristic of P. macularis (2) on both cultivars. Cleistothecia did not develop in any colonies. In addition, Nugget and Symphony plants were inoculated with a field population of P. macularis originating from cultivars lacking R6 in Oregon. These inoculations on Nugget did not develop powdery mildew whereas Symphony plants did. Non-inoculated controls remained free of powdery mildew. Results were identical in two additional experiments. The sequence of the mating type idiomorph, MAT1-1, was obtained to confirm identity of the pathogen as P. macularis as described previously (1). The sequences were identical among the three isolates obtained from the greenhouse in Washington and isolates of P. macularis obtained previously from Oregon and Washington. MAT1-2 idiomorph was not detected in the isolates collected. While R6-virulent strains have been detected previously in race characterization experiments, these strains have not caused widespread epidemics of powdery mildew. The increasing prevalence of virulent strains of P. macularis and outbreaks of powdery mildew on formerly resistant cultivars necessitates changes in breeding strategies and disease management efforts to minimize damage resulting from the disease. The distribution of virulent strains of the pathogen and susceptibility of formerly resistance cultivars to powdery mildew are currently under investigation. References: (1) B. Asalfet et al. Phytopathology 103:717, 2013. (2) R. Bélanger et al. The Powdery Mildews: a Comprehensive Treatise. APS Press, St. Paul, MN, 2002. (3) P. Darby. Brew Hist. 121:94, 2005. (4) C. Ocamb et al. Plant Dis. 83:1072, 1999.

Podosphaera macularis, the causal agent of hop powdery mildew, is a recurrent threat to hops in the Pacific Northwest because of the potential to reduce cone yield and quality. Early-season pruning is a common practice in hop production for horticultural reasons. Studies were conducted over a 3-year period in a commercial hop yard to quantify the effect of pruning method and timing on disease development, yield, and cone quality factors. A 4-week delay in pruning reduced the incidence of leaves with powdery mildew from 46 to 10% and cones from 9 to 1%, with the specific effect being season dependent. Pruning using chemical desiccants rather than by mechanical means had similar effects on disease levels on leaves. On cones, though, chemical pruning had a small but significant reduction in the incidence of powdery mildew compared with mechanical pruning. Cone yield, levels of bittering-acids, and color were not negatively affected in any individual year or cumulatively over three seasons when pruning treatments were applied repeatedly to the same plots during the study period. Delayed pruning may offer a low-cost means of reducing both the incidence of powdery mildew and early-season fungicide inputs in certain cultivars.

Canopy management is an important aspect of control of powdery mildew diseases and may influence the intensity of fungicide applications required to suppress disease. In hop, powdery mildew (caused by Podosphaera macularis) is most damaging to cones when infection occurs during bloom and the juvenile stages of cone development. Experiments were conducted over 3 years to evaluate whether fungicide applications could be ceased after the most susceptible stages of cone development (late July) without unduly affecting crop yield and quality when disease pressure was moderated with varying levels of basal foliage removal. In experimental plots of ‘Galena’ hop, the incidence of leaves with powdery mildew was similar whether fungicides were ceased in late July or made in late August. Disease levels on leaves were unaffected by the intensity of basal foliage removal, whereas the intensity of basal foliage removal interacted with the duration of fungicide applications to affect disease levels on cones. Similar experiments conducted in large plots of ‘Tomahawk’ hop in a commercial hop yard similarly found no significant impact on disease levels on leaves from either the duration of fungicide applications or intensity of basal foliage removal. In contrast, on cones, application of fungicides into August had a modest, suppressive effect on powdery mildew. There was also some evidence that the level of powdery mildew on cones associated with fungicide treatment was influenced by the intensity of basal foliage removal. When fungicide applications ceased in late July, there was a progressive decrease in the incidence of cones with powdery mildew with increasing intensity of basal foliage removal. Removing basal foliage two to three times allowed fungicide applications to be terminated in late July rather than late August without diminishing disease control on cones, yield, or cone quality factors. Thus, this study further establishes that fungicide applications made during the early stages of hop cone development have the strongest effect on suppression of powdery mildew on cones. The additive effect of fungicide applications targeted to the periods of greatest cone susceptibility and canopy management to reduce disease favorability may obviate the need for fungicide applications later in the season. This appears to be a viable strategy in mature hop yards of certain cultivars when disease pressure is not excessively high.

Host resistance, both quantitative and qualitative, is the preferred long-term approach for disease management in many pathosystems, including powdery mildew of hop (Podosphaera macularis). In 2012, an epidemic of powdery mildew occurred in Washington and Idaho on previously resistant cultivars whose resistance was putatively based on the gene designated R6. In 2013, isolates capable of causing severe disease on cultivars with R6-based resistance were confirmed in Oregon and became widespread during 2014. Surveys of commercial hop yards during 2012 to 2014 documented that powdery mildew is now widespread on cultivars possessing R6 resistance in Washington and Oregon, and the incidence of disease is progressively increasing. Pathogenic fitness, race, and mating type of R6-virulent isolates were compared with isolates of P. macularis lacking R6 virulence. All isolates were positive for the mating type idiomorph MAT1-1 and were able to overcome resistance genes Rb, R3, and R5 but not R1 or R2. In addition, R6-virulent isolates were shown to infect differential cultivars reported to possess the R6 gene and also the R4 gene, although R4 has not yet been broadly deployed in the United States. R6-virulent isolates were not detected from the eastern United States during 2012 to 2015. In growth chamber studies, R6-virulent isolates of P. macularis had a significantly longer latent period and produced fewer lesions on plants with R6 as compared with plants lacking R6, indicating a fitness cost to the fungus. R6-virulent isolates also produced fewer conidia when compared with isolates lacking R6 virulence, independent of whether the isolates were grown on a plant with or without R6. Thus, it is possible that the fitness cost of R6 virulence occurs regardless of host genotype. In field studies, powdery mildew was suppressed by at least 50% on plants possessing R6 as compared with those without R6 when coinoculated with R6-virulent and avirulent isolates. R6 virulence in P. macularis appears to be race specific and, at this time, imposes a measurable fitness penalty on the fungus. Resistance genes R1 and R2 appear to remain effective against R6-virulent isolates of P. macularis in the U.S. Pacific Northwest.

Mold identification at the species level in environmental samples is a major challenge. Molecular techniques have been widely used for fungal classification, but as most primers are genus-specific, it is laborious to identify unknown samples. In this study, a PCR-based method for the identification of mold at the species level was developed. Therefore, common sequencing primers and combinations of them, targeting specific DNA regions, were tested. Here we present a combination of eight primer pairs to identify mold within a single PCR run. The approach correctly identified mold of unknown species from samples taken at a local bakery, including Penicillium chrysogenum, Penicillium citrinum, Cladosporium sphaerospermum, Paecilomyces formosus, Rhizopus oryzae and Aspergillus niger. Results obtained from the PCR method were successfully validated by chromatographic mycotoxin and microscopy analysis. Findings highlight DNA barcoding as an appropriate tool for mold identification; however, its efficacy is essentially dependent on DNA quality and primer selection.

The hop powdery mildew fungus Podosphaera macularis persists from season to season in the Pacific Northwestern United States through infection of crown buds because only one of the mating types needed to produce the ascigerous stage is presently found in this region. Bud infection and successful overwintering of the fungus leads to the emergence of heavily infected shoots in early spring (termed flag shoots). Historical data of flag shoot occurrence and incidence in Oregon and Washington State during 2000 to 2017 were analyzed to identify their association with the incidence of powdery mildew, growers’ use of fungicides, autumn and winter temperature, and other production factors. During this period, flag shoots were found on 0.05% of plants evaluated in Oregon and 0.57% in Washington. In Oregon, the incidence of powdery mildew on leaves was most severe and the number of fungicide applications made by growers greatest in yards where flag shoots were found in spring. Similarly, the incidence of plants with powdery mildew in Washington was significantly associated with the number of flag shoots present in early spring, although the number of fungicide applications made was independent of flag shoot occurrence. The occurrence of flag shoots was associated with prior occurrence of flag shoots in a yard, the incidence of foliar powdery mildew in the previous year, grower pruning method, and, in Washington, winter temperature. A census of hop yards in the eastern extent of the Oregon production region during 2014 to 2017 found flag shoots in 27 of 489 yards evaluated. In yards without flag shoots, 338 yards (73.2%) were chemically pruning or not pruned, whereas the remaining 124 (26.8%) were mechanically pruned. Of the 27 yards with flag shoots, 22 were either chemically pruned or not pruned and 4 were mechanically pruned in mid-April, well after the initial emergence of flag shoots. The prevalence of yards with flag shoots also was related to thoroughness of pruning in spring (8.1% of yards with incomplete pruning versus 1.9% of yards with thorough pruning). A Bayesian logistic regression model was fit to the data from the intensively assessed yards in Oregon, with binary risk factors for occurrence of a flag shoot in the previous year, occurrence of foliar mildew in the previous year, and thoroughness of pruning in spring. The model indicated that the median and 95% highest posterior density interval of the probability of flag shoot occurrence was 0.0008 (0.0000 to 0.0053) when a yard had no risk factors but risk increased to 0.0065 (0.0000 to 0.0283) to 0.43 (0.175 to 0.709) when one to all three of the risk factors were present. The entirety of this research indicates that P. macularis appears to persist in a subset of chronically affected hop yards, particularly yards where spring pruning is conducted poorly. Targeted management of the disease in a subset of fields most at risk for producing flag shoots could potentially influence powdery mildew development regionwide.

During 2012, soil samples from commercial pistachio orchards in three major production regions include Rafsanjan (Kerman Province, center of Iran), Damghan (Semnan Province, north-central Iran) and Feyz-Abad (Khorasan-e Razavi Province, northeastern Iran), were assayed on Dichloran Rose-Bengal Chloramphenicol agar (DRBC) and Aspergillus flavus-parasiticus agar media to quantify populations of Aspergillus species from the section Flavi. The mean propagule density of Aspergillus species from the Flavi section [log10 (CFU/g soil)] was higher in Feyz-Abad (3.06, 2.88–3.24), compared to Damghan (2.55, 2.44–2.65) and Rafsanjan (2.40, 2.26–2.54). A. flavus (69.7, 65.3 and 57.9%), A. parasiticus (19.6, 25.4, and 29.3%), and A. nomius (10.7, 9.3, and 12.8%) were the predominant species in the regions of Rafsanjan, Damghan, and Feyz-Abad, respectively. There were significant differences among sclerotia producing isolates of A. flavus in the sampling regions (p < 0.05). The percentage of sclerotium-producing isolates of A. flavus from Rafsanjan (14.5%) was much lower than Damghan (39.5%) and Feyz-Abad (41.4%). The A. flavus isolates from Damghan, Rafsanjan, and Feyz-Abad were toxigenic at 53.7%, 61.6%, and 60.4%, respectively. In Rafsanjan, aflatoxin B1 (AFB1), and AFB1 + AFB2 (aflatoxin B2) ranged from 274 to 553 ppb (393±17.11) and 394 to 3745 ppb, respectively, while AFB1, and AFB1 + AFB2 ranged from 257 to 392 ppb (285±13.18) and 415 to 1658 ppb, respectively, in Damghan. We found 16 and 20 vegetative compatibility groups (VCGs) for 41 and 37 nit mutant producing isolates of A. flavus from Rafsanjan and Damghan, respectively. From Damghan the VCG diversity for A. flavus isolates was greater (54%) than from Rafsanjan (39%). Because there were a few number of sclerotium-producing isolates of A. flavus, we did not determine the relationships between sclerotium production with VCGs and/or geographical distribution in the three pistachio production regions. This study was the first to determine the strain and VCG diversity of A. flavus soil isolates from Iranian pistachio orchards.

The Aspergillus flavus population structure from maize kernels was examined. During 2011, samples were collected from two main grain maize production areas in Iran (Fars and Ardebil provinces), shortly before harvest. One-hundred nine A. flavus isolates were recovered on Dichloran Rose Bengal Chloramphenicole (DRBC) agar and Aspergillus flavus/parasiticus medium (AFPA) and grouped into morphotypes and Vegetative Compatibility Groups (VCGs) based on morphological (e.g. sclerotia production), physiological (e.g. aflatoxin-producing ability) and genetic criteria (e.g. heterokaryosis). In general, morphotype and VCG composition were highly dissimilar in both provinces. In total, 43.8% and 44.3% of A. flavus isolates from Ardebil and Fars, respectively, produced sclerotia. Sclerotia producers were identified as A. flavus L and S strain morphotypes in Ardebil (66.7% and 33.3%, respectively) and Fars (29.6% and 70.4%, respectively). Furthermore, 71 isolates (65.1%) were able to produce aflatoxin (Ardebil 40.8%, Fars 59.2%). The aflatoxin values were categorized into four different classes (< 10, 10-100, 100-1,000 and > 1,000 ppb). In total, 51 aflatoxin producing isolates of A. flavus (Ardebil n = 22, Fars n = 29) were assigned into 26 VCGs by complementation of nit auxotrophs on nitrate medium. None of the A. flavus isolates from Ardebil complemented with any isolates from Fars. Genetic diversity of A. flavus isolates was 59.1% and 41.8% for Ardebil and Fars, respectively. The different geographical adaptation and genetic make-up of A. flavus isolates may be due to different climatic conditions, soil types and crop sequences in both maize production areas.

Powdery mildew of sweet cherry in the Pacific Northwest (PNW) is caused by Podosphaera clandestina, which infects fruit and leaves. Disease symptoms are commonly observed near harvest and such infections are a major concern for offshore cherry markets due to the possibility of diseased cherries providing inoculum to establish the pathogen in a region where P. clandestina has not been reported. The present study was designed to identify periods at which fruit infections are most severe and to determine the efficacy of PNW’s industry postharvest handling practices on the survival of P. clandestina conidia on diseased fruit surfaces. Morphological characteristics and next-generation sequencing (NGS) platform was used to identify the causal agent of foliar and fruit powdery mildew found in the PNW. We combined qPCR and, the use of a viability dye, propidium monoazide (PMA), to differentiate between viable and dead (membrane-compromised) conidia. These tools helped to determine the viability of conidia in real-time. Conidia on the fruit surface naturally lost their viability within a few days of harvest. Postharvest treatments in the PNW include hydrocooling with chlorinated water and methyl bromide fumigation prior to export. These practices were extremely effective in deactivating all conidia on the fruit surface. The residual conidia recovered from the fruit surface after the postharvest treatments failed to establish consistent powdery mildew colonies on the susceptible host leaves; this confirms that diseased, treated cherries are unlikely to serve as a source of inoculum needed to establish the pathogen areas where it has not been documented. After postharvest treatments, regular sampling of conidia up to three weeks after harvest indicated that the conidia were non-viable. Results of this study indicated that the sweet cherry postharvest handling procedures in the PNW are effective for the elimination of the potential sources of P. clandestina inoculum associated with cherry fruit.

Humans and animals are exposed to aflatoxins, toxic carcinogenic fungal metabolites, through consumption of contaminated food and feed. Aspergillus flavus, the primary causal agent of crop aflatoxin contamination, is composed of phenotypically and genotypically diverse vegetative compatibility groups (VCGs). Molecular data suggest that VCGs largely behave as clones with certain VCGs exhibiting niche preference. VCGs vary in aflatoxin-producing ability, ranging from highly aflatoxigenic to atoxigenic. The prevalence of individual VCGs is dictated by competition during growth and reproduction under variable biotic and abiotic conditions. Agronomic practices influence structures and average aflatoxin-producing potentials of A. flavus populations and, as a result, incidences and severities of crop contamination. Application of atoxigenic strains has successfully reduced crop aflatoxin contamination across large areas in the United States. This strategy uses components of the endemic diversity to alter structures of A. flavus populations and improve safety of food, feed, and the overall environment.

Aflatoxins are highly carcinogenic mycotoxins frequently produced by Aspergillus flavus. Contamination of maize with aflatoxins imposes both economic and health burdens in many regions. Identification of the most important etiologic agents of contamination is complicated by mixed infections and varying aflatoxin-producing potential of fungal spe- cies and individuals. In order to know the potential importance of an isolate to cause a con- tamination event, the ability of the isolate to produce aflatoxins on the living host must be determined. Aflatoxin production in vitro (synthetic and natural media) was contrasted with in vivo (viable maize kernels) in order to determine ability of in vitro techniques to pre- dict the relative importance of causal agents to maize contamination events. Several media types and fermentation techniques (aerated, non-aerated, fermentation volume) were compared. There was no correlation between aflatoxin production in viable maize and pro- duction in any of the tested liquid fermentation media using any of the fermentation tech- niques. Isolates that produced aflatoxins on viable maize frequently failed to produce detectable (limit of detection ¼ 1 ppb) aflatoxin concentrations in synthetic media. Aflatoxin production on autoclaved maize kernels was highly correlated with production on viable maize kernels. The results have important implications for researchers seeking to either identify causal agents of contamination events or characterize atoxigenic isolates for biological control. Published by Elsevier Ltd on behalf of The British Mycological Society.

Aflatoxin contamination of crops is a world-wide problem. Lethal aflatoxicosis of humans has been associated with maize produced in Kenya for over three decades. The S strain morphotype of Aspergillus flavus was identified as the primary cause of aflatoxin contamination events occurring between 2004 and 2006 in Kenya. Because the S strain was first described in the U.S., it was suggested that the agent causing lethal levels of aflatoxins was introduced to Kenya with maize. DNA sequence comparisons among 68 S strain isolates from Kenya, the Americas, Asia, and Australia suggest the Kenyan isolates are distinct from those causing aflatoxin contaminations in the U.S. Analyses of 4.06 kb representing three loci from distinct chromosomes indicate that most S strain isolates from the U.S. resolved into a clade distinct from one containing the 30 Kenyan isolates. S strain isolates from Kenya were more closely related to the recently described species A. minisclerotigenes than to A. flavus. Furthermore, failure of the Kenyan isolates to produce G aflatoxins was attributed to a previously undescribed deletion in the cypA gene, suggesting that different deletion events led to loss of G aflatoxin production in S strain isolates from the U.S. and Kenya. Thus, although the Kenyan isolates have S strain morphology and produce large quantities of only B aflatoxins like A. flavus S strain isolates, these isolates are phylogenetically divergent from those described from other regions. The molecular characteristics of the Kenyan S strain isolates described herein are valuable tools to identify and track these highly aflatoxigenic fungi.

Aspergillus flavus has two morphotypes, the S strain and the L strain, that differ in aflatoxin-producing ability and other characteristics. Fungal communities on maize dominated by the S strain of A. flavus have repeatedly been associated with acute aflatoxin poisonings in Kenya, where management tools to reduce aflatoxin levels in maize are needed urgently. A. flavus isolates (n = 290) originating from maize produced in Kenya and belonging to the L strain morphotype were tested for aflatoxin-producing potential. A total of 96 atoxigenic isolates was identified from four provinces sampled. The 96 atoxigenic isolates were placed into 53 vegetative compatibility groups (VCGs) through complementation of nitrate non-utilizing mutants. Isolates from each of 11 VCGs were obtained from more than one maize sample, isolates from 10 of the VCGs were detected in multiple districts, and isolates of four VCGs were found in multiple provinces. Atoxigenic isolates were tested for potential to reduce aflatoxin concentrations in viable maize kernels that were co-inoculated with highly toxigenic S strain isolates. The 12 most effective isolates reduced aflatoxin levels by >80%. Reductions in aflatoxin levels caused by the most effective Kenyan isolates were comparable with those achieved with a United States isolate (NRRL-21882) used commercially for aflatoxin management. This study identified atoxigenic isolates of A. flavus with potential value for biological control within highly toxic Aspergillus communities associated with maize production in Kenya. These atoxigenic isolates have potential value in mitigating aflatoxin outbreaks in Kenya, and should be evaluated under field conditions.

Aims: To evaluate the potential role of fungal community structure in predisposing Kenyan maize to severe aflatoxin contamination by contrasting aflatoxin-producing fungi resident in the region with repeated outbreaks of lethal aflatoxicosis to those in regions without a history of aflatoxicosis. Methods and Results: Fungi belonging to Aspergillus section Flavi were isolated from maize samples from three Kenyan provinces between 2004 and 2006. Frequencies of identified strains and aflatoxin-producing abilities were assessed, and the data were analysed by statistical means. Most aflatoxin-producing fungi belonged to Aspergillus flavus. The two major morphotypes of A. flavus varied greatly between provinces, with the S strain dominant in both soil and maize within aflatoxicosis outbreak regions and the L strain dominant in nonoutbreak regions. Conclusions: Aspergillus community structure is an important factor in the development of aflatoxins in maize in Kenya and, as such, is a major contributor to the development of aflatoxicosis in the Eastern Province. Significance and Impact of the Study: Since 1982, deaths caused by aflatoxincontaminated maize have repeatedly occurred in the Eastern Province of Kenya. The current study characterized an unusual fungal community structure associated with the lethal contamination events. The results will be helpful in developing aflatoxin management practices to prevent future outbreaks in Kenya.

Aflatoxin contamination of tree nuts is a growing concern for pistachio producing countries. Development of competitive exclusion strategies through application of atoxigenic Aspergillus flavus isolates is a highly effective route of natural aflatoxin mitigation. Aflatoxin assays conducted on a high number of native A. flavus isolates are a first step to identify potential biological control isolates. Many cultural methods for the rapid and visual identification of atoxigenic A. flavus isolates have been described. The current study identified atoxigenic A. flavus isolates from Iranian pistachio orchards using and contrasting cultural, analytical and molecular methods. Ammonium vapour (AV) and fluorescence detection (FD), two rapid aflatoxin assays, were directly compared using various media preparations to screen 524 A. flavus isolates obtained from Iranian pistachio orchards. Percentages of false negatives were high using FD assays for all media preparations ranging from 13 to 15 %. This in contrast to AV assays. Here incidences of false negatives ranged from 0 % (using coconut agar medium) to 7.2 % (using potato dextrose agar). Aflatoxin-producing ability of all isolates was further confirmed using thin layer- and high-performance liquid chromatography. Sixty three atoxigenic A. flavus isolates were identified as atoxigenic in all assays. For these isolates, five loci across the aflatoxin biosynthesis cluster pathway were compared to identify genetic defects explaining atoxigenicity. Genetic deletions in at least one of five loci in the aflatoxin biosynthesis pathway were found for 97 % of isolates. Frequencies of atoxigenic strains ranged from 7.1 to 37.5 % with the lowest incidence detected in the Kerman province. Proper identification of atoxigenic isolates is considered a first step in the development of biological control strategies. Ability of identified isolates to competitively exclude aflatoxin-producing fungi has to be further investigated. © 2014 Koninklijke Nederlandse Planteziektenkundige Vereniging.

Despite clinical and laboratory evidence of perioperative hypercoagulability, there are no consistent data evaluating the extent, duration, and specific contribution of platelets and procoagulatory proteins by in vitro testing. We tested the hypothesis that the parallel use of standard and abciximab-cytochalasin D-modified thromboelastography (TEG) can assess 7 days' postoperative hypercoagulability and can estimate the independent contribution of procoagulatory proteins and platelets. Thromboelastograms were performed before surgery, at the end of surgery, 6 h after surgery, and on postoperative days 1, 2, 3, and 7; they were analyzed for the reaction time and the maximal amplitude (MA). We calculated the elastic shear modulus of standard MA (G(t)) and modified MA (G(c)), which reflect total clot strength and procoagulatory protein component, respectively. The difference was an estimate of the platelet component (G(p)). There was a 10% perioperative increase of standard MA, corresponding to a 50% increase of G(t) (P0.0001) and an 86%-90% contribution of the calculated G(p) to G(t). We conclude that serial standard and modified thromboelastography may reveal prolonged postoperative hypercoagulability and the independent contribution of platelets and procoagulatory proteins to clot strength.Postoperative hypercoagulability, occurring for at least 1 wk after major abdominal surgery, may be demonstrated by standard and modified thromboelastography. This hypercoagulability is not reflected by standard coagulation monitoring and seems to be predominantly caused by increased platelet reactivity.

Crops frequently contaminated by aflatoxins are important sources of revenue and daily nourishment in many portions of sub-Saharan Africa. In recent years, reports have associated aflatoxins with diminished human health and export opportunities in many African Nations. Aflatoxins are highly carcinogenic metabolites mainly produced by members of Aspergillus sect. Flavi. The current study examined aflatoxin-producing fungi associated with maize grain intended for human consumption in 18 sub-Saharan African countries. 4469 Aspergillus sect. Flavi isolates were obtained from 339 samples. The majority (75%) of isolates belonged to the L strain morphotype of A. flavus. Minor percentages were A. tamarii (6%), A. parasiticus (1%), and isolates with S strain morphology (3%). No A. bombycis or A. nomius isolates were detected. Phylogenetic analyses of partial sequences of the nitrate reductase gene (niaD, 1.3 kb) and the aflatoxin pathway transcription factor gene (aflR, 1.7 kb) were used to verify isolate assignments into species and lineages. Phylogenetics resolved S strain isolates producing only B aflatoxins into two lineages fully supported by sizes of deletions in the gene region spanning the aflatoxin biosynthesis genes cypA (aflU) and norB (aflF). One lineage was the A. flavus S strain with either 0.9 or 1.5 kb deletions. The second lineage, recently described from Kenya, has a 2.2 kb deletion. Taxa with S strain morphology differed in distribution with strain SBG limited to West Africa and both A. minisclerotigenes and the new lineage from Kenya in Central and East Africa. African A. flavus L strain isolates formed a single clade with L strain isolates from other continents. The sampled maize frequently tested positive for aflatoxins (65%), fumonisins (81%), and deoxynivalenol (40%) indicating the presence of fungi capable of producing the respective toxins. Percentage of samples exceeding US limits for total aflatoxins (regulatory limit), fumonisins (advisory limit), and deoxynivalenol (advisory limit) were 47%, 49%, 4%, respectively.

Aflatoxins are potent poisons that contaminate crops in warm regions worldwide and reduce health and economic welfare in several portions of Africa. Crops are contaminated in two phases: First, Aspergillus species infect crops during development; and second, after maturation contamination builds during exposure to warm humid conditions. Identification of the exact fungi causing contamination can provide clues to management strategies. Crops usually are infected by complex mixtures of aflatoxin-producing and closely related fungi. Among these are atoxigenic strains that produce no aflatoxins. In the United States atoxigenic strains are used to reduce contamination. Such technologies also may have value in Africa.

Maize contaminated with aflatoxins has been implicated in deadly epidemics in Kenya three times since 1981, but the fungi contaminating the maize with aflatoxins have not been characterized. Here we associate the S strain of Aspergillus flavus with lethal aflatoxicoses that took more than 125 lives in 2004.