Evaluation of food grade antioxidant formulation for sustained antifungal,antiaflatoxigenic and insecticidal activities on peanut conditioned at different water activities

The aim of this study was to investigate antifungal and insecticidal activity of two microencapsulated antioxidants: 2(3)-tert-butyl-4 hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT) against Aspergillus section Flavi and Oryzaephilus surinamensis (L.), a vector carrier of aflatoxigenic fungi on stored peanuts. Susceptibility of Aspergillus section Flavi, insects, and aflatoxin B₁ accumulation in sterile peanut kernels conditioned at two different water activities (a_w) (0.83 a_w and 0.95 a_w) was determined with different doses of antioxidant formulations (10, 20 and 30 mM) during 45 days. Moreover, Aspergillus section Flavi isolation frequency from live and dead insects was evaluated. The BHA formulation completely inhibited Aspergillus section Flavi development regardless of a_w and doses assayed. Antifungal effect of microencapsulated BHT was highly dependent on a_w, with 86–100% fungal inhibition at 20 and 30 mM, at the lowest a_w (0.83 a_w) and at the end of the experiment. No aflatoxin accumulation was detected in samples treated with the BHA formulation. In general, low levels of Aspergillus section Flavi were detected in dead insects. Our results show efficacy for 45 days, in addition microencapsulated BHT could be an alternative to control peanut pests in dry kernels.     

Application of essential oils in maize grain: impact on Aspergillus section Flavi growth parameters and aflatoxin accumulation

The antifungal activity of Pimpinella anisum L. (anise), Pëumus boldus Mol (boldus), Hedeoma multiflora Benth (mountain thyme), Syzygium aromaticum L. (clove), and Lippia turbinate var. integrifolia (griseb) (poleo) essential oils (EOs) against Aspergillus section Flavi was evaluated in sterile maize grain under different water activity (a_w) condition (0.982, 0.955, and 0.90). The effect of EOs added to maize grains on growth rate, lag phase, and aflatoxin B₁ (AFB₁) accumulation of Aspergillus section Flavi were evaluated at different water activity conditions. The five EOs analyzed have been shown to influence lag phase and growth rate. Their efficacy depended mainly on the essential oil concentrations and substrate water activity conditions. All EOs showed significant impact on AFB₁ accumulation. This effect was closely dependent on the water activity, concentration, and incubation periods. Important reduction of AFB₁ accumulation was observed in the majority of EO treatments at 11 days of incubation. Boldus, poleo, and mountain thyme EO completely inhibited AFB₁ at 2000 and 3000 μg g⁻¹. Inhibition of AFB₁ accumulation was also observed when aflatoxigenic isolates grew with different concentration of EOs during 35 days.     

Impact of volatile compounds generated by essential oils on Aspergillus section Flavi growth parameters and aflatoxin accumulation

BACKGROUND: The advantage of essential oils is their bioactivity in the vapour phase, a characteristic that makes them attractive as possible fumigants for stored grain protection. In this study the antifungal and antiaflatoxigenic effects of the volatile fractions of five essential oils (EOs) were evaluated by vapour contact on Aspergillus section Flavi isolates. RESULTS: In maize meal extract agar the volatile fractions of Pimpinella anisum L. (anise), Pëumus boldus Mol. (boldus), Hedeoma multiflora Benth. (mountain thyme), Lippia turbinata var. integrifolia (Griseb.) (poleo) and Syzygium aromaticum L. (clove) were able to decrease the growth rate and lag phase of aflatoxigenic isolates. Boldus EO showed the best antifungal effect on Aspergillus section Flavi growth rate. In sterilised maize grains, boldus and poleo EOs showed antifungal effects on growth rate and aflatoxin accumulation. The volatile fraction of boldus EO completely inhibited the growth of isolates at water activity (a_w) levels of 0.955, 0.930 and 0.900, while poleo EO showed this effect only at the lower a_w levels (0.930 and 0.900). All aflatoxigenic isolates showed reduced total aflatoxin accumulation in the presence of boldus EO under all a_w conditions. CONCLUSION: These findings clearly indicate that the volatile fraction of boldus EO could be used to control aflatoxigenic fungi in stored maize. Copyright © 2009 Society of Chemical Industry     

Effect of Pisum sativum fractions on the mortality and progeny production of nine stored-grain beetles

Yellow field pea (Pisum sativum L.) fractions that were mainly protein (50%), fibre (90%) or starch (85%) were obtained from a commercial pea mill and mixed with wheat kernels or wheat flour. Based on the mortality and the number of offspring produced, protein-rich pea flour was more toxic than fibre, which was more toxic than starch. For the protein-rich pea flour mixed with wheat kernels, the most sensitive insects were Sitophilus oryzae (L.), Sitophilus zeamais Motschulsky and Sitophilus granarius (L.), followed by Cryptolestes ferrugineus (Stephens) which was more sensitive than Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.). For the protein-rich pea flour mixed with wheat flour, Cryptolestes pusillus (Schönherr) was most sensitive, followed by C. turcicus (Grouvelle) and T. confusum (Jacquelin du Val), with T. castaneum being the most resistant. Although protein-rich pea flour did not kill adults to a great extent when mixed with flour, it reduced offspring production significantly. Again C. pusillus was the most sensitive, followed by T. confusum, with T. castaneum offspring being the most resistant. The insecticidal activity of pea fractions decreased after treated wheat kernels were held at 30 °C, 70% r.h. for 8 months. The potential of using pea fractions to control stored-product insects is discussed.     

A survey on distribution and toxigenicity of Aspergillus section Flavi in poultry feeds

Thirty-five samples of poultry feeds and corresponding raw materials (maize, soybean and meat meal) from a processing plant were analyzed to evaluate the distribution and toxigenicity of Aspergillus section Flavi isolates. Mycological analysis of the samples indicated the presence of five fungal genera (Aspergillus, Penicillium, Fusarium, Cladosporium, and Eurotium). Aspergillus flavus was the predominant species being present in 48.5% of the analyzed samples. Ninety-one isolates belonging to Aspergillus section Flavi were isolated; ninety were identified as A. flavus and only one as A. parasiticus. Fifty-seven isolates were capable of producing sclerotia, 41 were identified as L-type strains and 16 as type S. Fifty-seven percent of the isolates produced AFB₁ levels ranging from 0.05 μg/kg to 27.7 μg/kg whereas 86.8% produced CPA from 1.5 μg/kg to 137.8 μg/kg. L-strains produced from 0.05 to 14.8 μg/kg of aflatoxin and type S produced levels from 0.05 to 1.65 μg/kg. No significant differences in CPA production among S- and L-strains were observed. Sclerotial isolates produced AFB₁ levels ranging between 0.05 and 27.7 μg/kg and CPA levels from 3.8 to 47.3 μg/kg. More than half of the A. flavus isolates were able to produce AFB and CPA simultaneously. Twenty percent of the 35 samples were contaminated with aflatoxin B₁ whereas 34.3% were contaminated with CPA. The high rate of CPA producing isolates represents a potential risk of contamination with this toxin in poultry feeds.     

Multiplex PCR assay for the detection of aflatoxigenic and non-aflatoxigenic fungi in meju, a Korean fermented soybean food starter

Aflatoxins are toxic secondary metabolites produced commonly by Aspergillus flavus and Aspergillus parasiticus. In this study, the possibility of using multiplex PCR was investigated to speed up and specify the detection of aflatoxigenic Aspergillus species in meju, a traditional Korean fermented soybean food starter. Two different sets of three primers were designed specifically for the omtB, ver-1, aflR, and omtA genes present in the aflatoxin biosynthesis cluster. The optimized multiplex PCR showed that only aflatoxigenic Aspergillus species gave three band patterns in both primer sets. The detection limits were determined as 125 pg/μl for genomic DNA from aflatoxigenic A. parasiticus KCCM 35078, and 10⁵ spores/g of meju sample for DNA extracted directly from meju. A total of 65 Aspergillus isolates from meju were tested for the presence of aflatoxigenic fungi by the application of multiplex PCR, and were analyzed by TLC and HPLC for the aflatoxin production in the culture filtrates. Results showed a good correlation between the presence of the aflatoxin biosynthesis genes analyzed by multiplex PCR and aflatoxin production by TLC and HPLC. This suggests that this multiplex PCR method may provide an accurate and specific detection of aflatoxigenic Aspergillus species in fermented soybean foods.     

Protection of stored maize from insect pests using a two-component biological control method consisting of a hymenopteran parasitoid, Theocolax elegans, and transgenic avidin maize powder

The hymenopteran parasitoid, Theocolax elegans (Westwood), and transgenic avidin maize powder were tested to determine if their individual or combined use would protect stored grain from infestation by both internal and external insect pests. Small-scale tests were conducted in plastic jars containing 3 kg of non-transgenic maize. We tested treatments of 0.3% powdered avidin maize, the parasitoid wasp, and the combination of the parasitoid plus 0.3% powdered avidin maize. One pair each of Sitophilus zeamais Motschulsky, Tribolium castaneum (Herbst), and Cryptolestes ferrugineus (Stephens) was added to each jar. After 8 weeks, the entire contents of each jar were examined for adult insects. Control and avidin maize powders had no detrimental effects on the beneficial insect parasitoid T. elegans. The parasitoid suppressed populations of the internal feeder S. zeamais. The avidin maize powder treatment had no effect on S. zeamais because these larvae developed inside the maize kernels where no avidin maize powder was present. For S. zeamais, the combination treatment was not significantly different from the parasitoid treatment. In contrast, populations of the external feeder T. castaneum were not suppressed by the parasitoid but were suppressed by the avidin maize powder treatment. The parasitoid-avidin combination treatment produced the greatest percentage reduction for all three insect species and resulted in 78%, 94%, and 70% reductions in populations of S. zeamais, T. castaneum, and C. ferrugineus, respectively, when compared to the control treatment. The percentage reductions for the parasitoid treatment were 70%, 8%, and 20% for S. zeamais, T. castaneum, and C. ferrugineus, respectively. For the avidin maize powder treatment, populations of S. zeamais, T. castaneum, and C. ferrugineus were reduced by 10%, 85%, and 40%, respectively. The combination treatment of avidin maize powder plus the release of parasitoid wasps was superior to either treatment alone when applied to mixed populations of internal and external feeders.     

Effect of antioxidants and competing mycoflora on Fusarium verticillioides and F. proliferatum populations and fumonisin production on maize grain

This study was carried out to determine the temporal effect of the antioxidants butylated hydroxyanisol (BHA) and propyl paraben (PP) at doses of 500 and 1000 μg/g on the growth of Fusarium verticillioides and F. proliferatum inoculated on natural maize grain in the presence of the competing mycoflora and fumonisin production at 0.98 and 0.95 water activity (a_w) over a 28-day storage period. The reduction in the log colony forming units (CFU) of Penicillium, Aspergillus and Fusarium populations was 10-100 fold depending on dose of BHA or PP, a_w and time. However, the populations of all three groups were higher at 0.98 a_w than 0.95 a_w. BHA at 500 μg/g and 0.95 a_w reduced the fumonisin content by 82% after 7-14 days incubation, but at the end of the experimental period the reduction was only 32%. A higher reduction in the level of fumonisin produced (77%) was achieved with BHA at 1000 μg/g after 28 days. PP at 500 and 1000 μg/g decreased fumonisin production throughout the incubation period in the drier treatment, but at 0.98 a_w control of toxin production was only achieved after 7-14 days. The reduction in the fumonisin levels could be due to the combined effect of antioxidants, and the competing mycoflora, mainly Aspergillus and Penicillium species.     

Potential use of phenolic antioxidants on peanut to control growth and aflatoxin B1 accumulation by Aspergillus flavus and Aspergillus parasiticus

Food‐grade antioxidants: butylated hydroxyanisole (BHA), propyl paraben (PP) and butylated hydroxytoluene (BHT) (10 and 20 mmol g^?1) and all the mixtures of these chemicals were tested for inhibitory activity on the growth of and aflatoxin B₁ (AFB₁) accumulation by Aspergillus parasiticus and A. flavus on irradiated (7 kGy) peanut grains. Also, the influence of these treatments was evaluated in different water conditions (0.982, 0.955, 0.937a_w) at 11 and 35 days of incubation at 28 °C. Water activity (a_w) affected the fungal growth, no fungal development was observed at the highest stress water condition (0.937a_w). Butylated hydroxyanisole at 10 mmol g^?1 level and all the mixtures with PP and/or BHT were significantly effective (P = 0.05) in increasing lag phase and reducing growth rate and colony forming units per gram of peanut of both Aspergillus section Flavi strains and AFB₁ accumulation. The application of BHA at concentrations of 20 mmol g^?1 alone or with PP and/or BHT totally inhibited fungal growth at 11 and 35 days of incubation. The results suggest that the addition of these chemical mixtures on peanut grains at low levels has potential to impact synergically on the control of Aspergillus section Flavi. Copyright © 2007 Society of Chemical Industry     

Resistance of Tripsacorn to Sitophilus zeamais and Oryzaephilus surinamensis

One strategy that has been used to find germplasm for developing improved plant varieties is to test ancestral germplasm for the desired traits. Although the progenitors of commercial maize are not known, a hybrid (called Tripsacorn) developed from a perennial teosinte, Zea diploperennis, and eastern gamagrass, Tripsacum dactyloides, resembles the earliest known samples of primitive domesticated maize. We tested resistance of whole Tripsacorn to the primary storage pest (primary storage pests can infest intact kernels) the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), and resistance of ground Tripsacorn to the secondary storage pest (secondary pests usually cannot infest intact kernels) the sawtoothed grain beetle, Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae). Tripsacorn was immune to attack by S. zeamais. The weevils were unable to lay eggs in the Tripsacorn, and we hypothesized that the hardness of the fruitcase was responsible for lack of weevil oviposition. Oryzaephilus surinamensis were able to complete immature development on ground Tripsacorn, but duration of development was longer and weight of emerged adults was less than for beetles developing on wheat. Hardness of the fruitcase may have been a primitive mechanism of defense against insects and other pests, but probably would not be an acceptable trait in commercial varieties. It remains to be determined whether the possible antibiotic effect demonstrated in ground Tripsacorn would be a useful trait in commercial maize hybrids.     

Efficacy of powder and essential oil from Chenopodium ambrosioides leaves as post-harvest grain protectants against six-stored product beetles

Powder and essential oil obtained from dry ground leaves of Chenopodium ambrosioides were tested under laboratory conditions (25±1°C, 70-75% r.h.) for their ability to protect grains from damage by six insect pests, Callosobruchus chinensis, C. maculatus, Acanthoscelides obtectus, Sitophilus granarius, S. zeamais and Prostephanus truncatus. The insects were reared and tested on whole maize grain for S. zeamais and P. truncatus, whole wheat for S. granarius, green peas for C. chinensis, mung bean for C. maculatus and white bean for A. obtectus. The powder prepared from dry leaves of C. ambrosioides was mixed with grains at different dosages ranging from 0.05-0.80% (wt/wt) for C. chinensis, C. maculatus and A. obtectus and from 0.8-6.4% (wt/wt) for S. granarius, S. zeamais and P. truncatus. The dosage of 0.4% killed more than 60% of all the bruchids 2 days after treatment, while a dosage of 6.4% induced total mortality of S. granarius and S. zeamais within the same exposure time. All levels of the dry ground leaf concentrations inhibited F1 progeny production and adult emergence of the tested insects. The dosage of 0.2 μl/cm² of the essential oil killed 80-100% of the beetles within 24 h except C. maculatus and S. zeamais, where this dosage induced only 20% and 5% mortality, respectively. These results indicate a scientific rationale for the use of this plant in grain protection by local communities in the western highlands of Cameroon.     

Fumigant toxicity of essential oils from three Thai plants (Zingiberaceae) and their major compounds against Sitophilus zeamais, Tribolium castaneum and two parasitoids

Fumigant toxicity of essential oils from rhizomes of Alpinia conchigera, Zingiber zerumbet, Curcuma zedoaria and their major compounds; camphene, camphor, 1,8-cineole, α-humulene, isoborneol, α-pinene, β-pinene and terpinen-4-ol was investigated with adults of Sitophilus zeamais, Tribolium castaneum, Anisopteromalus calandrae and Trichogramma deion larvae. The last two insects are parasitoids commonly used to control stored-product weevils and moths. The trial was evaluated at 0, 37, 74, 148, 296, 444, 593 μL/L in air after 12, 24 and 48 h for S. zeamais, T. castaneum and A. calandrae, and 24 h for T. deion. Alpinia conchigera oils were toxic to S. zeamais, T. castaneum and T. deion, while the other two plant oils had low toxicity. Adults of S. zeamais and T. castaneum were more susceptible to A. conchigera oils than their eggs, larvae or pupae. Sitophilus zeamais adults (LC₅₀ 85 μL/L in air) were slightly more tolerant of A. conchigera oils than T. castaneum (LC₅₀ 73 μL/L in air) after 48 h exposure. Synthetic essential oils, a mixture of pure compounds in the same ratios of the extracted essential oils, were tested with S. zeamais and T. castaneum adults. Synthetic essential oils were more toxic than the extracted essential oils to both insects. Zingiber zerumbet oils (LC₅₀ 26 μL/L in air) and C. zedoaria oils (LC₅₀ 25 μL/L in air) were significantly more toxic to adults of A. calandrae than A. conchigera oils (LC₅₀ 37 μL/L in air) whereas T. deion larvae were more sensitive to A. conchigera oils (LC₅₀ 62 μL/L in air) than Z. zerumbet and C. zedoaria oils (LC₅₀ > 593 μL/L in air). Tribolium castaneum was more susceptible than S. zeamais to the eight pure compounds. Terpinen-4-ol was highly toxic to both insects.     

Effect of CO2 modified atmosphere packaging on aflatoxin production in maize infested with Sitophilus zeamais

The weevil Sitophilus zeamais (Motschulsky), the maize weevil, is a pest of stored maize that can cause feeding damage and lead to the proliferation of toxigenic fungi. The application of modified atmospheres with a high concentration of CO₂ is an alternative method for the control of S. zeamais and the inhibition of fungal growth. The objectives of the study were to determine the effect of S. zeamais infestation, grain damage and grain moisture content on aflatoxin production by Aspergillus flavus on maize, and the impact of high CO₂ modified atmosphere packaging on pest infestation and aflatoxin production. Mycotoxin production was only recorded when maize was infested with S. zeamais and had A. flavus inoculum. However, production of mycotoxins was not recorded when the maize was mechanically damaged and stored at 18% moisture content, indicating that the biological activity of the insect was determinant in the production of mycotoxins. The high CO₂ modified atmosphere packaging tested (90% CO₂, 5% O₂ and 5% N₂) prevented mycotoxin production.     

Aflatoxins and ochratoxin A in stored barley grain in Spain and impact of PCR-based strategies to assess the occurrence of aflatoxigenic and ochratoxigenic Aspergillus spp

Contamination of barley by moulds and mycotoxins results in quality and nutritional losses and represents a significant hazard to the food chain. The presence of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) and ochratoxin A (OTA) in stored barley in Spain has been studied. Species-specific PCR assays were used for detection of Aspergillus flavus, A. parasiticus, A. ochraceus, A. steynii, A. westerdijkiae, A. carbonarius and A. niger aggregate in mycotoxin-positive barley samples at different incubation times (0, 1 and 2 days). Classical enumeration techniques (CFU/g) in different culture media for evaluation of Aspergillus in sections Flavi, Circumdati and Nigri were also used. One hundred and five barley kernel samples were collected in Spanish grain stores from 2008 to 2010, and analyzed using a previously optimized method involving accelerated solvent extraction, cleanup by immunoaffinity column, liquid chromatographic separation, post-column derivatization with iodine and fluorescence detection. Twenty-nine samples were contaminated with at least one of the studied mycotoxins. AFB1, AFB2, AFG1, AFG2, and OTA were detected in 12.4%, 2.9%, 4.8%, 2.9%, and 20% of the samples, respectively. Aflatoxins and OTA co-occurred in 4.8% of the samples. Maximum mycotoxin levels (ng/g) were 0.61 (AFB1), 0.06 (AFB2), 0.26 (AFG1), 0.05 (AFG2), and 2.0 (OTA). The results of PCR assays indicated the presence of all the studied species, except A. westerdijkiae. The PCR assays showed high levels of natural contamination of barley with the studied species of Aspergillus which do not correspond to the expected number of CFU/g in the cultures. These results suggest that a high number of non-viable spores or hyphae may exist in the samples. This is the first study carried out on the levels of aflatoxins and OTA in barley grain in Spain. Likewise, this is the first report on the presence of aflatoxigenic and ochratoxigenic Aspergillus spp. in barley grain naturally contaminated with those mycotoxins using a species-specific PCR approach.     

Efficacy of diflubenzuron plus methoprene against Sitophilus oryzae and Rhyzopertha dominica in stored sorghum

The efficacy of diflubenzuron (1 mg kg⁻¹)+methoprene (1 mg kg⁻¹) against Sitophilus oryzae (L.) and Rhyzopertha dominica (F.) in sorghum was evaluated in a silo-scale trial in southeast Queensland, Australia. Sorghum is normally protected from a wide range of insects by mixtures of grain protectants. The chitin synthesis inhibitor diflubenzuron was evaluated as a potential new protectant for S. oryzae in combination with the juvenile hormone analogue methoprene, which is already registered for control of R. dominica. Sorghum (ca 200 t) was treated after harvest in 2000 and assessed for treatment efficacy and residue decline during 6.5 months storage. The reproductive capacity of S. oryzae and R. dominica was greatly reduced in bioassays of treated sorghum throughout the trial, and efficacy remained relatively stable during the trial. An initial exposure of S. oryzae adults to treated sorghum for 2 weeks reduced F₁ progeny production of all strains by 80.8-98.8%, but a second exposure of 4 weeks reduced F₁ progeny production by 98.5-100%. In addition, the reproductive capacity of any S. oryzae progeny produced was greatly reduced. Exposure of R. dominica adults to treated sorghum for 2 weeks reduced F₁ progeny production of all strains by 99.6-100%, including a methoprene-resistant strain. The results indicate that S. oryzae or R. dominica adults invading sorghum treated with diflubenzuron (1 mg kg⁻¹)+methoprene (1 mg kg⁻¹) would be incapable of producing sustainable populations.     

Screening of Chinese medicinal herbs for bioactivity against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst)

Extracts of 40 species of Chinese medicinal herb from 32 different botanical families were screened for contact, fumigant and feeding-deterrent activities against the two stored-grain insects Sitophilus zeamais and Tribolium castaneum. Thirty Chinese medicinal herbs exhibited insecticidal or feeding-deterrent activities against the two species of insects. Extracts of Artemisia argyi, Dictamnus dasycarpus, Evodia rutaecarpa, Litsea cubeba, Narcissus tazetta var. chinensis, Polygonum aviculare, Rhododendron molle, Sophora flavescens, Stemona sessilifolia, Tripterygium wilfordii, and Torreya grandis were most active.     

Aflatoxigenic fungi and aflatoxin in cocoa

This paper reports the occurrence of aflatoxigenic fungi and the presence of aflatoxins in 226 cocoa samples collected on Brazilian farms. The samples were taken at various stages of fermentation, drying and storage. A total of 819 potentially aflatoxigenic fungi were isolated using Dichloran 18% Glycerol agar after surface disinfection, and identified by standard techniques. The ability of the fungi to produce aflatoxins was determined using the agar plug technique and TLC. The presence of aflatoxins in cocoa samples was determined by HPLC using post-column derivatization with bromide after immunoaffinity column clean up. The aflatoxigenic fungi isolated were Aspergillus flavus, A. parasiticus and A. nomius. A considerable increase in numbers of these species was observed during drying and storage. In spite of the high prevalence of aflatoxigenic fungi, only low levels of aflatoxin were found in the cocoa samples, suggesting the existence of limiting factors to the accumulation of aflatoxins in the beans.     

Predominant mycobiota and aflatoxin content in Brazil nuts

The Brazil nut (Bertholletia excelsa) is an economically important product to the Brazilian Amazon. Currently, its marketing is being affected by the high incidence of aflatoxins (AF) produced by potentially aflatoxigenic fungi associated with its seeds. In this context, the purpose of this study was to determine which part of the nut contributes to contamination by aflatoxins and to identify the mycobiota in Brazil nut samples. Unshelled and shelled nuts were analyzed by measuring the total count of filamentous fungi (Aspergillus sections Flavi, Nigri and Circumdati) in sanitised and non-sanitised treatments. The isolates identified as Aspergillus section Flavi, the major producers of AF, were plated for determination of their aflatoxigenic potential. To perform the AF analysis, samples of Brazil nuts were treated separately. The AF from the shell and kernel were extracted by chloroform and analysed by the HPLC-FD system in isocratic mode. The Aspergillus section Flavi count was 21.67% lower. The production of AF by the isolated fungi was 30% for sanitised and 23.8% for non-sanitised samples. The concentrations obtained of AFB₁ and AFG₁ were higher than those of AFB₂ and AFG₂. The AFB₁ concentrations of shelled nuts and shell samples were 35.0 and 1.78???g/kg, respectively. AFB₂ and AFG₂ were detected only in shelled nut samples. The HPLC-FD analysis presented limits of detection (LOD) and quantification (LOQ) of 0.2 and 0.4???g/kg, respectively.     

Insect population dynamics in commercial grain elevators

Data were collected in 1998-2002 from wheat stored in commercial grain elevators in south-central Kansas. Bins at these elevators had concrete walls and were typically 6-9 m in diameter and 30-35 m tall. A vacuum-probe sampler was used to collect grain samples in the top 12 m of the wheat in each bin. The primary insect species found in the wheat samples were: Cryptolestes ferrugineus, Rhyzopertha dominica, and Tribolium castaneum. In the top 3.7 m of grain, R. dominica, C. ferrugineus, T. castaneum and Sitophilus oryzae made up 44, 36, 19 and 1% of the insects found in the samples, respectively. From 3.8 to 12.2 m, R. dominica, C. ferrugineus, T. castaneum and S. oryzae were present at 84, 8, 8, and 1%, respectively. The most prevalent species also changed over time. In June, the start of wheat harvesting and storage in Kansas, insect density was low in the bins. At this time, C. ferrugineus was the most common insect, and it was found mostly in the top grain sample (0-1.2 m). In September through November, C. ferrugineus and R. dominica were at similar densities; however, from February to March, R. dominica was more common.Generally, insect density was greatest at the top and decreased with grain depth. Very few insects were found in samples collected from greater than 12 m (most of the bins contained grain to depths of 24-36 m). Insect density for all species increased rapidly from June through October. During this period less than 20% of the bins had economically significant insect densities (>2 insects/kg). From October until February, the average insect density remained fairly constant but it was greatly reduced in April, May, and June. Bins that had insect densities >2 insects/kg tended to be located adjacent to other heavily infested bins.     

A simple technique to assess compounds that are repellent or attractive to stored-product insects

We have developed a simple and rapid technique that mimics storage conditions, and determines if products are repellent or attractive to stored-product insects. The technique determines the response of insects to potential repellents and attractants by measuring their movement from grain. The technique used a device consisting of a perforated cup (2 mm perforations) that holds 200 g of wheat. A Petri dish and cup collected the insects as they left the wheat. Several natural products were tested for repellency: diatomaceous earth (DE), ground peas (Pisum sativum), protein-rich pea flour, pea starch, and pea fibre. Adult insects of three species were tested: the rice weevil, Sitophilus oryzae, the red flour beetle, Tribolium castaneum, and the rusty grain beetle, Cryptolestes ferrugineus. DE at 0.01% was repellent to all insects tested. Pea fibre, pea protein, and ground pea at 1% caused increased emigration of C. ferrugineus from the wheat. Pea starch did not affect movement out of the grain for all three insects. Only pea fibre and ground pea increased the movement of T. castaneum out of the grain. For S. oryzae, there were no differences after 1 h, but after 24 h both pea protein and ground pea increased movement out of the grain. Several potential attractants were placed outside the grain and the emigration out of the grain noted. For R. dominica, the commercial R. dominica pheromone increased the emigration of insects from the grain; R. dominica adults on broken grain enclosed in a ventilated vial in the collection jar also increased emigration, but not as much as the synthetic pheromone. The commercial Tribolium pheromone did increase movement out of the grain for T. castaneum, but the other treatments were no different from the control. None of the potential attractants increased the movement of S. oryzae from the grain. The implications of this work are discussed with reference to controlling and sampling stored-product insect pests.