Effect of non-aflatoxigenic strains of Aspergillus flavus on aflatoxin contamination of pre-harvest peanuts in fields in China

Aflatoxin contamination of peanuts is one of the most concerns in peanut production in China. Applying non-aflatoxigenic Aspergillus flavus strains, based on competitive exclusion, has been proved to be a promising strategy to reduce aflatoxin contamination in pre-harvest peanuts. Two non-aflatoxigenic A. flavus strains collected in China, which have been proved effectively reducing aflatoxin in the laboratory, were mixed with high aflatoxin producer to the soil in peanut growing season. The two non-aflatoxigenic strains significantly (P ​< ​0.05) reduced aflatoxin contamination in peanut kernels under both normal and drought stresses in two fields. Compared to control, the total aflatoxin (sum of aflatoxin B₁ and B₂) was reduced 26.7–99.12% in field 1, and 84.96–99.33% in field 2. The aflatoxin was reduced 84.96–99.33% under drought stress in two fields. The present study indicated the non-aflatoxigenic A. flavus strains could be potential biocontrol agents for reducing aflatoxin contamination under field condition.     

Aflatoxin in maize,a multifaceted answer of Aspergillus flavus governed by weather,host-plant and competitor fungi

Aspergillus section Flavi is able to produce aflatoxins (AFs) in vitro down to 0.85 a_w with a potential maximum occurring between 0.95 and 0.99 a_w, while in the field AFs increased significantly with kernel humidity below 0.95 a_w. In order to clarify this apparent discrepancy, a 3-year field trial with artificial inoculation of maize ears with Aspergillus flavus strains was organised. The co-occurrence of Aspergillus section Flavi and Gibberella fujikuroi species complex (Gfsc) was observed. The incidence of A. flavus was significantly influenced by the year and negatively related to Gfsc incidence. In 2012, when the highest temperature and the lowest rain were registered, aflatoxin B₁ (AFB₁) content was the highest and a_w < 0.95 in kernels was measured early throughout the growing season. In 2013 and 2014, the temperature was lower and rain more abundant, and a_w decreased below 0.95 only close to harvest and AFB₁ contamination was limited. The possibility of describing/predicting reasonably well a_w dynamic based on temperature, (degree day) was confirmed. With a_w > 0.95, a positive correlation between AFB₁ production rate and a_w was found, but a negative correlation resulted with a_w < 0.95. Other factors than a_w play a role, but a_w = 0.95 should be considered as an indicator of suitable conditions for rapid aflatoxin accumulation.     

Aflatoxin contamination of corn under different agro-environmental conditions and biocontrol applications

Biological control of the fungus Aspergillus flavus has been shown to be effective in reducing aflatoxin contamination in corn. This study compared field application of a bioplastic-based formulation for delivering atoxigenic A. flavus isolates in Northern Italy and the Mississippi Delta.Due to an extremely hot and dry summer at the Italy site in 2012, aflatoxin contamination was approximately seven times higher than in 2011. In 2011, and 2012, application of bioplastic granules inoculated with the atoxigenic isolate A. flavus NRRL 30797 at 15 and 30 kg ha⁻¹ resulted in a reduction of aflatoxin contamination by 67.2 ± 4.1% and 94.8 ± 5.3%, respectively. The higher application rate was also effective when soil abundance of A. flavus was artificially increased by applying contaminated corn residues. At the Mississippi site, summer 2012 was also hot and dry, with high levels of aflatoxin contamination. In fields planted with non-Bt or Bt hybrids, application of biocontrol granules inoculated with A. flavus NRRL 30797 or NRRL 21882 at 30 kg ha⁻¹ reduced aflatoxin contamination to up to 89.6%. Field experiments on two continents showed that bioplastic-based A. flavus formulations markedly reduced aflatoxin contamination under different agro-environmental conditions and infestation intensities.     

Fungal species isolated from peanuts in major Kenyan markets: Emphasis on Aspergillus section Flavi

A survey was conducted in Nairobi, Nyanza and Western provinces in Kenya between March and July 2009 with 1263 peanut products sampled out of which 705 samples underwent microbial analysis. The study aimed at determining the incidence of fungal species – emphasis on Aspergillus section Flavi – associated with peanut products. A 0.5 kg representative sample was obtained from each surveyed vendor and the colony forming units (CFU) of fungal species determined. The samples were also analyzed for total aflatoxin level while isolates of Aspergillus flavus and Aspergillus parasiticus were screened for production of aflatoxin B1, B2, G1 and G2. Eight fungal species were detected in the samples and were in decreasing order of CFU/g of sample: A. flavus S-strain (467), A. flavus L-strain (341), Penicillium spp. (326), Aspergillus niger (156), Aspergillus tamari (27), Aspergillus alliaceus (21), A. parasiticus (10), and Aspergillus caelatus (5). The overall incidence of A. flavus S-strain in samples from Nairobi was 92 and 1425% higher than samples from Nyanza and Western regions, respectively. The combined incidence of A. flavus and A. parasiticus was varied significantly (p ≤ 0.05) with peanut product: peanut flour (69%), shelled raw peanuts (53%), spoilt peanuts (49%), boiled podded peanuts (45%), podded peanuts (39%), peanut butter (31%), fried peanuts (22%) and roasted peanuts (20%). Seventy three percent of A. flavus and A. parasiticus isolates produced at least one of the aflatoxin types, with 66% producing aflatoxin B1. The total aflatoxin level among peanut products ranged from 0 to 1629 μg/g; and there was a positive correlation (r = 0.2711) between the incidence of A. flavus and A. parasiticus, and total aflatoxin level. The high incidence of aflatoxin producing fungi in peanuts traded in Kenyan markets implies a risk of aflatoxin contamination, highlighting the need for stakeholders to promote sound practices at all stages of the peanut value chain in order to minimize market access by non-complying products.     

Occurrence and distribution of aflatoxin contamination in groundnuts (Arachis hypogaea L) and population density of Aflatoxigenic Aspergilli in Malawi

Groundnut (Arachis hypogaea L.) is susceptible to pre- and post-harvest infections by Aspergillus spp. Aflatoxin B₁ (AFB₁), is the contaminant produced by the fungus in infected grains posing a threat to human and animal health. This paper reports of a study undertaken in Malawi to determine the occurrence and distribution of Aflatoxigenic Aspergilli in the soil and AFB₁ contamination in groundnuts. A total of 1397 groundnut samples collected from farm homesteads, local markets, warehouses and shops in 2008 and 2009 were analyzed for AFB₁ contamination using the enzyme linked immunosorbent assay (ELISA), and A. Aspergilli population densities in 1053 soil samples collected from the same sites were estimated using serial dilutions plated on A. Aspergilli medium. Farmer socio-economic profile information was also collected to determine relationships to AFB₁ contamination. The results revealed 46% and 23% of the total samples, from 2008 to 2009, respectively, had AFB₁ contamination levels greater than 4 ppb, and those above 20 ppb were 21% for 2008 and 8% for 2009, respectively. Fitted smooth curve relationships show that there is a clear increase in the chance of groundnut contamination when the population density of A. Aspergilli in the soil increased beyond 3000 (log (cfu) > 8). The measured level of A. Aspergilli in soil varied by location, as well as ecologies within location. Low-altitude ecologies, which were warmer and experienced low precipitation levels, had the highest densities of A. Aspergilli, whereas cooler high-altitude ecologies had the lowest density of these fungi. Similarly high AFB₁ contamination, was recorded across the country with 11–28% of all samples collected from the warm low to mid-altitude ecologies recording contamination ≥20 ppb and low contamination (2–10% of samples) in the mid to high altitude cool ecologies. From a crop management perspective, this study also suggests that both less experienced and older farmers were more likely to produce groundnuts contaminated with aflatoxin. These findings have implications in the design of intervention strategies to avoid short- and long-term human health effects from aflatoxin exposure.     

Geographical distribution of Aspergillus flavus in peanut harvest period in China

In order to grasp the distribution of Aspergillus fiavus in the soil of peanut production areas in China, A. fiavus biomarkers were tested on 555 soil samples from 37 sampling points in 17 provinces, peanut fields in four agroecological zones(Southern area, Yangtze River Basin, Northern area, Northeast area). The results showed that(1) the cultivation amount of A. fiavus per gram of soil in the Yangtze River Basin is 1.30 times that of the southern area, 1.56 times that of the northern area, and...     

Characterization of laboratory pyrimethanil-resistant mutants of Aspergillus flavus from groundnut in China

To assess the potential risk of resistance development in Aspergillus flavus to pyrimethanil, five highly pyrimethanil-resistant (Pyr^R) mutants (RF > 996.2) were obtained after UV-mutagenesis and tested for fitness parameters and aflatoxin B1 production. All five mutant strains had mycelial growth rate, sporulation and aflatoxin production similar to or even higher than the wild-type parent strain, which indicated that pyrimethanil possesses a high risk in the development of resistance in A. flavus. Comparing the sequences of four key enzymes cystathionine β-lyase (CBL), cystathionine γ-synthase (CGS), methionine sulfoxide reductase (MsrB), and sulfate permease (SP2) involved in the biosynthesis and metabolism of methionine and sulfate assimilation revealed that no amino acid difference was found between the mutant and wild-type parent strains, suggesting that the four enzymes might not be related to the anilinopyrimidines (APs) resistance in A. flavus.     

Drought,pod yield,pre-harvest Aspergillus infection and aflatoxin contamination on peanut in Niger

Soil moisture and soil temperature affect pre-harvest infection with Aspergillus flavus and production of aflatoxin. The objectives of our field research in Niger, West Africa, were to: (i) examine the effects of sowing date and irrigation treatments on pod yield, infection with A. flavus and aflatoxin concentration; and (ii) to quantify relations between infection, aflatoxin concentration and soil moisture stress. Seed of an aflatoxin susceptible peanut cv. JL24 was sown at two to four different sowing dates under four irrigation treatments (rainfed and irrigation at 7, 14 and 21 days intervals) between 1991 and 1994, giving 40 different ‘environments’. Average air and soil temperatures of 28–34 °C were favourable for aflatoxin contamination. CROPGRO-peanut model was used to simulate the occurrence of moisture stress. The model was able to simulate yields of peanut well over the 40 environments (r² = 0.67). In general, early sowing produced greater pod yields, as well as less infection and lower aflatoxin concentration. There were negative linear relations between infection (r² = 0.62) and the average simulated fraction of extractable soil water (FESW) between flowering and harvest, and between aflatoxin concentration (r² = 0.54) and FESW in the last 25 days of pod-filling. This field study confirms that infection and aflatoxin concentration in peanut can be related to the occurrence of soil moisture stress during pod-filling when soil temperatures are near optimal for A. flavus. These relations could form the basis of a decision-support system to predict the risk of aflatoxin contamination in peanuts in similar environments.     

Improvement in biocontrol activity of Bacillus subtilis UTB1 against Aspergillus flavus using gamma-irradiation

Bacillus subtilis UTB1, a biocontrol bacterium isolated from Iranian pistachio nuts, has revealed to have antagonistic activity against aflatoxin-producing Aspergillus flavus R5. The strain UTB1 produces lipopeptide compounds and is able to degrade aflatoxin B1. In this study, a random mutagenesis generated using different doses of gamma irradiation (0.1–3 KGy) was applied on B. subtilis UTB1 to improve its antagonistic activity against A. flavus R5. Five hundred bacterial colonies were selected randomly after irradiation, and their effects against A. flavus R5 were assessed in a plate assay. Forty-five colonies (9%) exhibited higher inhibition activity as compared to the non-irradiated wild type. Eight colonies out of the 45 were selected based on different polymorphism patterns obtained by repetitive element sequence polymorphism-PCR (ERIC and BOX) analyses; six of which could significantly inhibit the fungal growth utilizing washed cells and cell-free supernatants as compared to the parental strain. According to thin-layer chromatograms, the production of lipopeptides including surfactin, fengycin and iturin families increased in these six mutants. A considerable inhibition of the fungal growth was observed using bioautography analysis, which associated with iturins production. A. flavus sporulation and aflatoxin content decreased significantly in pistachio nuts treated with mutants M419 and M464 as compared to the strain UTB1. These results suggest that both mutants M419 and M464 could be promising biocontrol candidates against A. flavus in pistachio nuts.     

Heritability of,and genotypic correlations between,aflatoxin traits and physiological traits for drought tolerance under end of season drought in peanut (Arachis hypogaea L.)

More rapid progress in breeding peanut for reduced aflatoxin contamination should be achievable with a better understanding of the inheritance of, aflatoxin trait and physiological traits that are associated with reduced contamination. The objectives of this study were to estimate the heritability of aflatoxin traits and genotypic (r_G) and phenotypic (r_P) correlations between drought resistance traits and aflatoxin traits in peanut. One hundred-forty peanut lines in the F_4:6 and F_4:7 generations were generated from four crosses, and tested under well-watered and terminal drought conditions. Field experiments were conducted under the dry seasons 2006/2007 and 2007/2008. Data were recorded for biomass (BIO), pod yield (PY), drought tolerance traits [harvest index (HI), drought tolerance index (DTI) of BIO and PY, specific leaf area (SLA), and SPAD chlorophyll meter reading (SCMR)], and aflatoxin traits [seed infection and aflatoxin contamination]. Heritabilities of A. flavus infection and aflatoxin contamination in this study were low to moderate. The heritabilities for seed infection and aflatoxin contamination ranged from 0.48 to 0.58 and 0.24 to 0.68, respectively. Significant correlations between aflatoxin traits and DTI (PY), DTI (BIO), HI, biomass and pod yield under terminal drought conditions were found (r_P = −0.25** to 0.32**, r_G = −0.57** to 0.53**). Strong correlations between SLA and SCMR with A. flavus infection and aflatoxin contamination were also found. Positive correlations between SLA at 80, 90, and 100 DAP and aflatoxin traits were significant (r_P = 0.13** to 0.46**, r_G = 0.26** to 0.81**). SCMR was negatively correlated with aflatoxin traits (r_P = −0.10** to −0.40**, r_G = −0.11** to −0.66**). These results indicated that physiological-based selection approaches using SLA and SCMR might be effective for improving aflatoxin resistance in peanut.     

Ear rot,aflatoxin accumulation,and fungal biomass in maize after inoculation with Aspergillus flavus

Aflatoxin, a toxin produced by the fungus Aspergillus flavus Link: Fries, occurs naturally in maize (Zea mays L.). Aflatoxin is a potent human carcinogen and is also toxic to livestock, pets, and wildlife. When contaminated with aflatoxin, the value of maize grain is markedly reduced. This investigation was conducted to compare ear rot, aflatoxin accumulation, and fungal biomass in maize single crosses with varying degrees of resistance to aflatoxin accumulation and to determine the relative importance of general combining ability (GCA) and specific combining ability (SCA) in the inheritance of resistance to ear rot, aflatoxin accumulation, and fungal biomass. Eight germplasm lines with different levels of resistance to aflatoxin accumulation were used as parents of a diallel cross. The cross was evaluated for visible ear rot, aflatoxin accumulation, and A. flavus infection in the grain. A. flavus infection was determined by quantitative real-time polymerase chain reaction (qRT-PCR) assays. Both GCA and SCA were significant sources of variation in the inheritance of the three traits although GCA accounted for a greater portion of the variation among single crosses. The interactions of GCA and SCA with years were highly significant for aflatoxin accumulation, but not significant for A. flavus infection. Estimates of GCA effects were highly significant for both reduced A. flavus infection and reduced aflatoxin accumulation for Mp313E, Mp715, and Mp717. Conversely, GCA effects associated with GA209 were significant for reduced levels of A. flavus infection and ear rot, but high levels of aflatoxin accumulation. Mp313E, Mp715, and Mp717 should be useful in breeding programs targeting both reduced levels of fungal infection and aflatoxin accumulation.     

Antifungal and antiaflatoxigenic efficacy of Caesulia axillaris Roxb. essential oil against fungi deteriorating some herbal raw materials, and its antioxidant activity

The study deals with evaluation of antifungal and antiaflatoxigenic Caesulia axillaris Roxb. essential oil (EO) against herbal raw materials deteriorating fungi and its free radical scavenging activity. During mycoflora analysis these herbal raw materials were found to be severely contaminated by different fungi and aflatoxins. A total of nine different fungal species were isolated from three herbal raw materials. Aspergillus flavus LHPtc was recorded as the highest aflatoxin B1 producing strain. EOs of some plants were tested for their fungitoxicity against the toxigenic strain A. flavus LHPtc, and C. axillaris EO was found as potent fungitoxicant. C. axillaris EO was chemically characterized through GC-MS analysis which depicted the presence of 18 compounds, dl-limonene and Euasarone being the major components. The EO exhibited broad spectrum of fungitoxicity against fungi causing postharvest deterioration of herbal raw materials. At 1.0 μl ml⁻¹ the oil showed complete inhibition of fungal growth and aflatoxin B₁ production was inhibited at 0.8 μl ml⁻¹. Free radical scavenging activity of the oil was also recorded by 2,2-diphenyl-1-picrylhydrazyl assay, and its IC₅₀ value was found 18 μl ml⁻¹. The safety limit of the EO was determined in terms of LD₅₀ on mice, which was 9166.6 μl kg⁻¹, suggesting its non mammalian toxicity. The EO of C. axillaris may be recommended as a plant based preservative in enhancement of shelf life of herbal raw materials by preventing their lipid peroxidation as well as biodeterioration due to fungal and aflatoxin contamination.     

Association between aflatoxin contamination and drought tolerance traits in peanut

The current study investigates the association between drought tolerance traits and aflatoxin contamination in peanut grown under long-term drought. Two field experiments were conducted at Khon Kaen University, Thailand using a split–split plot design with three drought stress levels as main plots, 11 genotypes as sub-plots, and two soil inoculations of Aspergillus flavus treatments as sub-sub-plots. The effects of temperature, soil moisture and A. flavus population on kernel colonization and aflatoxin contamination, and drought tolerance traits viz. specific leaf area (SLA) and root length density (RLD) were measured. The results demonstrated that elevated soil temperatures and reduced soil moisture, favored aflatoxin production. Drought in combination with higher levels of A. flavus inoculum load in the soil resulted in an increase in the fungal populations in the soil which in turn resulted in increased kernel colonization and subsequent aflatoxin contamination. A combination of SLA and RLD, and kernel colonization had a significant influence on aflatoxin contamination under drought conditions in both seasons (r = 0.73** and 0.76**). The results revealed that drought tolerance traits (SLA and RLD) could be contributing to resistance to aflatoxin contamination suggesting that a combination of SLA, RLD and kernel colonization could be used as selection criteria in selecting parents for aflatoxin resistance.     

Antioxidant Capacity and Antimutagenic Activity of Natural Oleoresin from Greenhouse Grown Tomatoes (<Emphasis Type="Italic">Lycopersicon esculentum</Emphasis>)

Natural oleoresins rich in lycopene were obtained from two varieties of tomato (Zedona and Gironda) and their nutraceutical potential (antioxidant and antimutagenic capacity) was evaluated. Both oleoresins had a high content of lycopene, 58.33 ± 1.67 mg/g (Zedona) and 63.97 ± 0.80 mg/g (Gironda). The antioxidant activity (AA) of the oleoresins by β-carotene method were 56.4–74.5% (Zedona) and 51–72.8% (Gironda), while when using the free radical stable 2,2-diphenyl-picryl-hydrazyl (DPPH) method, the antiradical activity (ARA) was determined to be 18.2–32.7% (Zedona) and 16.6–26.7% (Gironda) for the concentrations tested that of 200–400 μM equivalents of lycopene. The antimutagenic activity of the oleoresins was tested against aflatoxin B₁ (AFB₁) using the microsuspension assay, both varieties had a very high antimutagenic potential against AFB₁ (60–66%).These results suggest the NCRT can be taken advantage to obtaining rich oleoresin in lycopene with a nutraceutical value.     

Southwestern corn borer damage and aflatoxin accumulation in conventional and transgenic corn hybrids

Southwestern corn borer (Diatraea grandiosella Dyar) is a major pest of corn (Zea mays L.) in the southern United States. In addition to the direct yield losses caused by southwestern corn borer, larval feeding on developing ears provides a site for fungi to enter the ear. Aspergillus flavus Link: Fries infection and the subsequent accumulation of aflatoxin in corn grain are major limitations to profitable corn production in the southern United States. This investigation was conducted to determine the effectiveness of transgenic corn hybrids expressing the δ-endotoxin insecticidal (CryIAb) proteins isolated from Bacillus thuringiensis (Bt) in reducing southwestern corn borer damage and aflatoxin accumulation. Ear damage and aflatoxin accumulation were compared among 10 pairs of conventional nonBt and transgenic Bt corn hybrids following infestation with southwestern corn borer and inoculation with A. flavus using kernel-wounding and nonwounding techniques. Both nonBt and Bt hybrids exhibited high levels of aflatoxin accumulation when inoculated with a kernel-wounding technique. When inoculated with a non-wounding technique and infested with southwestern corn borer, aflatoxin accumulation was significantly higher in nonBt than Bt hybrids. Aflatoxin accumulation was also significantly higher for nonBt hybrids inoculated with A. flavus and infested with southwestern corn borer than for hybrids that were only inoculated with A. flavus. Southwestern corn borer larval establishment was significantly higher on nonBt hybrids than on Bt hybrids. Larval survival was extremely low on the Bt hybrids. The results of this investigation indicate that these Bt hybrids should be effective in reducing aflatoxin contamination in areas where high southwestern corn borer infestations occur. The reduced levels of aflatoxin accumulation associated with Bt hybrids are likely a consequence of reduced insect damage rather than resistance to A. flavus infection or aflatoxin accumulation per se.     

Screening,identification and antagonistic effect of antagonistic bacteria JTFM1001 against aflatoxin contamination in corn

Aflatoxin is a strong carcinogenic and toxic fungal toxin produced by Aspergillus flavus and other Aspergillus species,and can seriously threaten the health of consumers,thus becoming a global concern.Corn,as an important oil and economic crop,is highly susceptible contaminated by aflatoxin.In this study,antagonistic bacteria with strong inhibitory effect on aflatoxin were screened to provide support for the treatment aflatoxin contamination control in corn.Ten strains which have strong antagonistic effects against A.flavus were isolated from healthy corn from different corn producing areas in China.Among them,the antagonistic bacteria JTFM1001 through corn kernels in vivo and field experiment,the inhibition effect of aflatoxin contamination reached above 70%and 55%,respectively.And the strain was identified as Bacillus subtilis based on its morphological,physiological and biochemical characteristics and phylogenetic analysis of 16S rDNA.In addition,our data showed that it can colonize in the rhizosphere and survive for a long time,forming the dominant flora,with broad application prospect.Finally,we were surprised to find that the antibacterial metabolites secreted by the antagonistic bacteria was one of the mechanisms of its inhibition of A.flavus and aflatoxin.This will provide us with new ideas and perspectives on the effective prevention and control of aflatoxin contamination in corn and corn oil.     

Diallel analysis of aflatoxin accumulation in maize

Since its discovery in numerous feedstuffs, aflatoxin, a carcinogenic compound produced by the fungus Aspergillus flavus Link ex Fries, has caused much concern among consumers and producers alike. This toxin poses a serious economic threat to maize (Zea mays L.) producers of the southeastern and midwestern regions of the United States. Efforts to identify maize germplasm that is resistant to aflatoxin accumulation and to investigate the genetic basis of this resistance have been undertaken at numerous research institutions. The objectives of this study were to (1) evaluate aflatoxin accumulation in grain harvested from maize inbred lines and a diallel cross among these lines, (2) determine the importance of general and specific combining abilities in inheritance of resistance to aflatoxin accumulation, and (3) estimate general and specific combining ability effects associated with resistance to aflatoxin accumulation in the inbred lines and crosses among them. Eight inbred lines and a diallel cross of the maize lines were inoculated with an A. flavus spore suspension 12–14 d after silk emergence. Following harvest, aflatoxin content was determined from samples of grain. Statistical analyses performed using SAS general linear models (GLM) and DIALLEL-SAS indicated that general and specific combining ability were significant sources of variation in the inheritance of resistance to aflatoxin accumulation. The inbred line Mp313E, which was developed and released as a source of resistance to aflatoxin contamination, had significantly lower aflatoxin accumulation than other lines. Mo18W exhibited excellent general combining ability for reduced aflatoxin accumulation when crossed with the other lines. Both Mp313E and Mo18W could be useful in breeding programs to develop aflatoxin-resistant maize hybrids. Mp339, SC212M, and Ab24E demonstrated aflatoxin susceptibility as both inbreds and in single crosses.     

Indole Diterpenoids and Isocoumarin from the Fungus,Aspergillus flavus,Isolated from the Prawn,Penaeus vannamei

Two new indole-diterpenoids (1 and 2) and a new isocoumarin (3), along with the known β-aflatrem (4), paspalinine (5), leporin B (6), α-cyclopiazonic acid (7), iso-α-cyclopiazonic acid (8), ditryptophenaline (9), aflatoxin B₁ (10), 7-O-acetylkojic acid (11) and kojic acid (12), were isolated from the fermentation broth of the marine-derived fungus, Aspergillus flavus OUCMDZ-2205. The structures of Compounds 1–12 were elucidated by spectroscopic analyses, quantum ECD calculations and the chemical method. New Compound 1 exhibited antibacterial activity against Staphylococcus aureus with a MIC value of 20.5 μM. Both new Compounds 1 and 2 could arrest the A549 cell cycle in the S phase at a concentration of 10 μM. Compound 1 showed PKC-beta inhibition with an IC₅₀ value of 15.6 μM. In addition, the absolute configurations of the known compounds, 4–6 and leporin A (6a), were also determined for the first time.     

Effect of short term ingestion ofTelfaria occidentalis (fluted pumpkin) on the liver

Four groups of female mice (18–22g) were fed for at least 10 days on a control diet or diets with addedTelfaria occidentalis (10% w/w) vegetables. Group 1 were the control animals. Group 2 were fed the vegetable diet alone. Group 3 were fed the control diet and injected with aflatoxin B₁(AFB₁, 0.35 mg/kg/day) in the last 3 days. Group 4 were fed with the vegetable diet and similarly treated for 3 days with AFB₁. No effects attributable to treatment were encountered in body weight, food intake, water intake and relative liver weight (g/100 g body weight). Decreases (17–40%) in PB sleeping time in the vegetable fed animals were not significant (P>0.05) in comparison to the control. Microsomal proteins increased by 2 fold in group 2 but were apparently normal in groups 3 and 4. In group 2 the activities of liver microsomal glucose-6-phosphatase and NADH-cytochrome c reductase increased but those of NADPH-cytochrome c reductase and aniline hydroxylase decreased. Increases were observed in the activities of all of these enzymes in groups 3 and 4. The highest increases were observed in glucose-6-phosphatase activity in mice fed the vegetables and also injected with AFB₁.     

Sensitivity and cross-resistance patterns to demethylation-inhibiting fungicides in California populations of Alternaria alternata pathogenic on pistachio

Alternaria late blight (ALB) caused by Alternaria spp. is an annual production concern for pistachio growers in California. In this study, isolates of Alternaria alternata collected from California pistachio orchards between 1998 and 2003 (population A, n = 46) and in 2010 (population B, n = 38) prior to the registration and use of sterol demethylation-inhibiting (DMI) fungicides for ALB control, were tested for their sensitivity to difenoconazole, propiconazole, and tebuconazole using an in vitro mycelial growth assay. The 50% effective dose (EC₅₀) values for isolates in population A to difenoconazole, propiconazole, and tebuconazole, respectively, ranged from 0.02 to 0.82 μg/mL (mean = 0.17), 0.25–5.73 μg/mL (mean = 1.29), and 0.02–2.76 μg/mL (mean = 0.96), and from 0.02 to 0.93 μg/mL (mean = 0.21), 0.13–5.36 μg/mL (mean = 1.25), and 0.17–5.47 μg/mL (mean = 1.03) for those in population B. Among 65 isolates collected in 2012 from orchards with very limited exposure to difenoconazole, propiconazole or metconazole (population C, n = 65), the EC₅₀ values for tebuconazole ranged from 0.02 to 2.99 μg/mL (mean = 0.51), while EC₅₀ values for difenoconazole of 55 of these isolates varied from 0.01 to 0.44 μg/mL (mean = 0.06). The DMI-exposed population C was found to be on average two to four times less sensitive compared with the populations A and B. Pearson correlation analyses of EC₅₀ values for the three fungicides showed significant positive correlations between the sensitivities of tebuconazole and propiconazole (P < 0.01, r = 0.32), tebuconazole and difenoconazole (P < 0.01, r = 0.26), and difenoconazole and propiconazole (P < 0.01, r = 0.40). Results indicate a clear difference in fungicide intrinsic activity, with difenoconazole being more active than the two other DMIs. Although wide variations in sensitivities for tebuconazole and propiconazole were observed in the non- and DMI-exposed Alternaria populations, efficacy of DMI products has been excellent against ALB. The data collected here will be crucial for the rational use of DMIs in fungicide spray programs for ALB management and serve as a reference to detect any shifts in A. alternata sensitivity to these fungicides in subsequent years as they become more frequently used in California pistachios.