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.     

The effect of repellents on penetration into packaging by stored-product insects

Two known repellents of stored-product insects, DEET and neem, were compared to protein-enriched pea flour, defatted protein-enriched pea flour, and pea protein extract for their efficacy at reducing penetration and invasion by several common stored-product insects: Sitophilus oryzae (L.), Tribolium castaneum (Herbst), Cryptolestes ferrugineus (Stephens), and Oryzaephilus surinamensis (L.). The methods of preparation of pea extract affected the penetration of S. oryzae. Protein-enriched pea flour, DEET and neem reduced the penetration of S. oryzae, but defatted protein-enriched pea flour and pea protein extract did not. The number of S. oryzae, T. castaneum, C. ferrugineus and O. surinamensis entering pierced paper envelopes that contained wheat and were treated with DEET was reduced by 99%, 86%, 97% and 91%, respectively. Neem was less effective than DEET in reducing penetration and invasion of insects. Protein-enriched pea flour did not prevent insects entering pierced envelopes.     

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.     

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.     

Effects of developmental stage,cold acclimation and diet on the cold tolerance of three species of Cryptolestes (Coleoptera: Laemophloeidae)

Flat grain beetles (Coleoptera: Laemophloeidae) are common stored-product insect pests in Canada, infesting cereals in grain bins, equipment and end products in flour mills. We studied the cold tolerance of the three most common flat grain beetles: Cryptolestes ferrugineus, Cryptolestes turcicus and Cryptolestes pusillus, by measuring the survival at −10 °C and supercooling point (SCP) for different life stages (egg, young larva, old larva, pupa and adult) reared on flour mixed with brewer’s yeast. Probit analysis was used to estimate the lethal time for 50 and 95% mortality. This was done with non-acclimated individuals (only held at 30 °C) or cold-acclimated individuals (held at 18, 10 and 5 °C, for 1 week/temperature). In general, adults were the most cold-hardy stage for each of the species. Acclimated insects were anywhere from no increase in cold tolerance to 14-fold more cold-tolerant than the corresponding non-acclimated stage and species. Cryptolestes ferrugineus was most cold-tolerant species (58 d at −10 °C to reach 95% mortality for acclimated adult), C. turcicus was the next most cold-tolerant, (39 d) and C. pusillus was the least cold-tolerant (11 d). The cold tolerance of adults reared on three diets was measured both for acclimated and non-acclimated insects. The adults reared on grain diet (whole wheat kernels, cracked wheat kernels and wheat germ (90:5:5 mass ratio) were the most cold-tolerant, adults reared on white-wheat flour and brewer’s yeast diet (95:5 mass ratio) had the next highest cold tolerance followed by the adults reared on 100% white-wheat flour. Supercooling point (SCP) of insects ranged from −20.6 to −26.7 °C. In general, acclimated insects had slightly lower SCP than non-acclimated insects.     

Effects of Cry1F and Cry34Ab1/35Ab1 on storage pests

Two crystalline protoxins from Bacillus thuringiensis (Bt), Cry1Fa1 and Cry34Ab1/Cry35Ab1 (Cry1F, Cry34/35Ab1), were evaluated for efficacy against lepidopteran and coleopteran storage pests. Cry1F was tested against the lepidopterans Sitotroga cerealella (Angoumois grain moth) and colonies of Plodia interpunctella (Indian mealmoth) that are susceptible or resistant to Bt Cry1Ab and Cry1Ac toxins, Bt subspecies entomocidus, and the commercial formulation Dipel^®. Cry1F was also tested against the coleopterans Cryptolestes pusillus (flat grain beetle) and Tribolium castaneum (red flour beetle). Cry34/35Ab1 was tested against S. cerealella, C. pusillus, and T. castaneum, and against additional coleopteran storage pests, including Tenebrio molitor (yellow mealworm), Trogoderma variabile (warehouse beetle), Oryzaephilus surinamensis (sawtoothed grain beetle), Rhyzopertha dominica (lesser grain borer), and Sitophilus oryzae (rice weevil). Strains of Bt-susceptible or -resistant P. interpunctella generally were more sensitive to Cry1A protoxin or toxin than either Cry1F protoxin or Dipel. Despite difficulties with the bioassay of S. cerealella larvae, the data suggest that Cry1F and Cry34/35Ab1 caused increased larval mortality, and a developmental delay was observed and no pupae emerged with 0.9% Cry1F. Neither Cry1F nor the corn rootworm-active toxin Cry34/35Ab1 significantly affected the biological parameters of the coleopteran species evaluated.     

Toxic compounds in essential oils of coriander, caraway and basil active against stored rice pests

Essential oils, distilled from seeds of Coriander sativum and Carum carvii and from leaves of five different varieties of Ocimum basilicum, were fractionated by column chromatography and tested in the laboratory for volatile toxicity against three stored rice pests (Sitophilus oryzae, Rhyzopertha dominica and Cryptolestes pusillus). The active fractions were analyzed by GC-MS. Coriander contained linalool (1617 ppm of the oil) as the main product active against the three pests. Camphor-rich fractions (over 400 ppm) were very toxic to R. dominica and C. pusillus. The caraway profile included carvone and limonene as expected but (E)-anethole, generally regarded as a minor product in the essential oil of this species, was also a major component, being present at 365 ppm. Carvone was the most effective (972 ppm) monoterpenoid against S. oryzae. In addition, (E)-anethole at 880 ppm was toxic to R. dominica while vapors of limonene (1416 ppm) and fenchone-rich (554 ppm) fractions killed adults of C. pusillus only. Three major essential oil profiles were present in the five varieties of O. basilicum analyzed: methyl eugenol/estragole, estragole and estragole/linalool chemotypes. The abundance of components had a strong influence on the outcome of the bioassays. Fractions, where combinations of products occurred with or without other minor compounds, were often more toxic than any one compound alone.     

Grain loss caused by Tribolium castaneum, Sitophilus oryzae and Acanthoscelides obtectus in stored durum wheat and beans treated with Beauveria bassiana

The effect of the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin (Hyphomycete) on the losses caused to durum wheat and beans by storage insects was investigated. Grains were infested with Tribolium castaneum (Herbst), Sitophilus oryzae (L.) and Acanthoscelides obtectus (Say). Beauveria bassiana was produced in inoculated autoclaved rice. The spore formulation (ground rice and B. bassiana) was applied to grain (wheat or bean) and shaken to evenly cover the grain. Adults of T. castaneum or S. oryzae were added to wheat and adults of A. obtectus to bean. Five replicates were set up for each treatment and controls (milled rice but no conidia). The insecticidal effect of B. bassiana was tested by measuring the fresh weight and weight loss of grains after four months of storage. Wheat grains infested with S. oryzae without the conidia was significantly more damaged by weevils than grains treated with B. bassiana. The mean fresh weight of grains with the conidia was significantly greater (18.4%) than the corresponding mean without the fungus when S. oryzae were present. Percentage weight loss decreased by 81.5% and was significantly smaller than the loss from the untreated grain. Significant differences were not found in the fresh weight of seed infested with T. castaneum or A. obtectus in treated or untreated grain nor in the percentage weight loss of grains infested with these insects, with and without B. bassiana.     

Feeding bioassay for stored-product insect pests using an encapsulated food source

A procedure that was developed to encapsulate liquid and semiliquid diets was used to encapsulate dry diet for use in a feeding bioassay for beetles that are pests of stored products. Vacuum was used to form Parafilm^® into numerous 6 mm diameter wells. The wells were filled with clean sand (control) or ground dry dog food (test), and the Parafilm^® sealed to produce individual pellets. A single pellet was then placed in the center of a 9 cm diameter Petri dish and feeding activity of groups of ten adults of Sitophilus oryzae (L.), the rice weevil; Oryzaephilus surinamensis (L.), the sawtoothed grain beetle; and Tribolium castaneum (Herbst), the red flour beetle, was tested. The number of insects on the encapsulated pellet and the amount of food or sand scattered were checked hourly for the first 8 h of the study and after 24 h. Few insects were observed on pellets containing sand and little or no sand was observed scattered outside of the pellet, so the presence of dog food in the pellet was needed for insects to feed on the pellet. In tests with encapsulated dog food, the amount of food scattered provided a better quantitative measure of feeding than the number of insects on the pellet. Insects that were starved for 48 h caused greater amounts of food scatter than insects starved for 24 h prior to the test. In direct comparisons among all three species, T. castaneum responded the most slowly and the bioassay may be improved by increasing the amount of time starved. Sitophilus oryzae responded very quickly and the entire pellet was essentially consumed within the first 4-5 h of the study. The encapsulated diets provide a promising method to evaluate feeding behavior of stored-product insect pests.     

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.     

Detection of eggs of stored-product insects in flour with staining techniques

Chemical stains were investigated with the aim of developing a method for detecting eggs of stored-product insects in refined wheat flour (white flour), so that eggs and flour particles could be distinguished easily. Eggs of the rust-red flour beetle, Tribolium castaneum, the saw-toothed grain beetle, Oryzaephilus surinamensis and the rice moth, Corcyra cephalonica were detected by staining with 0.1% bromocresol green or 1:1 mixture of bromocresol green+0.5% orange G solution after acid digestion of the flour particles. In contrast to the standard AACC method of staining with iodine solution where both eggs and flour particles were stained brownish-yellow or yellow, bromocresol green alone or in mixture with orange G, differentially stained the insect eggs and the flour particles distinct orange and green, respectively. Eggs in flour could also be detected using 0.05% acid fuchsin but both eggs and flour particles were stained uniformly pink.     

Impact of physical and biological factors on susceptibility of Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) to new formulations of hydroprene

Three separate experiments were conducted to: (1) evaluate two new commercial formulations (202-080 and 202-084) of the insect growth regulator hydroprene, (2) determine residual efficacy of hydroprene-treated concrete stored at different environmental conditions, and (3) assess the impact of accumulated flour on residual efficacy. In the first test, late instars of Tribolium castaneum (Herbst), the red flour beetle, and Tribolium confusum (du Val), the confused flour beetle, were exposed on concrete treated with hydroprene. At 40% relative humidity (r.h.), there was no difference between species regarding the percentage of individuals that stopped development in the larval stage (arrested larvae), but at 75% r.h. there were more arrested T. castaneum than T. confusum in all treatments except the low rate of formulation 202-084. No adult T. castaneum lived after emergence (live adults) at either relative humidity, but the percentage of live adult T. confusum ranged from 1.0% to 41.0%, depending on treatment. In the second test, late instars of T. confusum were exposed at 6 and 12 weeks post-treatment on concrete treated with the two experimental formulations and stored under different environmental conditions. At 6 weeks there was no difference between formulations. At 12 weeks, fewer live adults and more dead emerged adults with gross morphological deformities were found on concrete treated with formulation 202-084 and stored at 32°C, 75% r.h. compared to other treatment combinations. In the final experiment, wheat flour was added to treated concrete for 5 weeks before the bioassays were conducted with late-instar T. confusum. There were few live adults produced in the initial bioassays, and dead adults with gross morphological deformities ranged from 83.1% to 97.6%. However, in bioassays conducted with late-instar larvae at 6 weeks, most adults eventually emerged with few deformities. The presence of the flour apparently compromised residual control and the hydroprene was no longer effective. In summary, the new hydroprene formulations were equivalent to the registered product Gentrol^®. Tribolium confusum was less susceptible than T. castaneum, and residual control of T. confusum on a clean surface without flour lasted about 6-12 weeks.     

Interspecific associations among stored-grain beetles

Recent increases in prices of raw grain, including wheat, will reduce action thresholds for insect damage and therefore justify more research into management practices and understanding of pest ecology in stored grain. Compared to most other habitats, natural or man-made, a filled grain silo constitutes a unique and fairly homogeneous habitat in which food availability for many grain-feeding insects is unlimited. A fundamental aspect of stored-grain insect ecology is a better understanding of associations among common beetle species. We analyzed the densities of three important stored-grain beetle species, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae), and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) in wheat samples collected in 1999–2001 from 129 grain silos in Kansas. The beetles studied here are highly mobile, and the number of insects in each grain sample is a result of the beetles' preference for favorable micro-environmental conditions and possibly of intra- and interspecific associations. In general, the number of T. castaneum in a grain sample increased as the number of R. dominica increased, but the number of C. ferrugineus was not correlated with the number of R. dominica. The densities of both T. castaneum and R. dominica decreased as the number of C. ferrugineus increased. Cryptolestes ferrugineus and T. castaneum can be predators and the species composition of insects in a grain sample may be modified by predation. As T. castaneum populations increased, so did R. dominica but not C. ferrugineus. Our analysis of the species composition in grain samples is discussed in an ecological context.     

Penetration of polyethylene sheets coated with protein-enriched pea flour solution by two stored-product insects

Adults of Sitophilus oryzae (L.) and Rhyzopertha dominica (F.) penetrated polyethylene sheets (0.0635 mm thickness) coated with protein-enriched pea flour solution (water pH 8.5-9.0) at 0 (control), 1 and 5% levels, but did not penetrate the sheets coated with a 10% concentration. The potential of using protein-enriched pea flour in the manufacture of insect-proof factory impregnated plastic sheeting is discussed.     

Laboratory assessment of insecticidal effectiveness of natural zeolite and diatomaceous earth formulations against three stored-product beetle pests

The insecticidal effectiveness of two natural zeolite formulations (Minazel plus and Minazel), applied to wheat at selected rates of 0.25, 0.50 and 0.75 g/kg, and a diatomaceous earth formulation (DE) (Protect-It™), applied at the recommended rates of 0.15 g/kg for Sitophilus oryzae, 0.20 g/kg for Rhyzopertha dominica and 0.30 g/kg for Tribolium castaneum, were tested under laboratory conditions (24 ± 1 °C temperature and 45 ± 5% relative humidity). The highest adult mortality was observed after the longest exposure period of 21 days and 7 days of recovery, when all three zeolite dosage rates and the recommended DE dosage caused 97-100% mortality of S. oryzae and 94-100% of T. castaneum. On the other hand, 100% mortality was not achieved in any test variant involving R. dominica; the highest (about 92%) was detected for DE, while 52% and 79% mortality was achieved with the zeolites at the highest rate of 0.75 g/kg. Progeny reduction by >90% was achieved after 21 days of contact of all three beetle pests with DE-treated wheat, while the same level of reduction was achieved for S. oryzae and T. castaenum only after contact with the highest rate of the zeolite product, Minazel. Thus the two zeolite formulations are comparable to diatomaceous earth in controlling adult S. oryzae, R. dominica and T. castaneum, but only the Minazel formulation could effectively protect wheat from attack by S. oryzae or T. castaneum, and only with a higher rate of application than for the DE formulation.     

Effect of suboptimal temperatures and sublethal CO2 levels on multiplication of Tribolium castaneum (Coleoptera: Tenebrionidae), alone or competing with Cryptolestes ferrugineus (Coleoptera: Cucujidae)

Groups of adults of Tribolium castaneum alone, or in combination with adults of Cryptolestes ferrugineus, were exposed to suboptimal temperatures (15°C, 20°C, and 25°C) and sublethal CO₂ levels (2%, 5%, and 10%) in dry (12%, wet mass basis) and damp (15%, wet mass basis) stored wheat in the laboratory, to investigate effects on population size. The mean adult numbers in single- and mixed-species tests were positively correlated with higher temperature and moisture content and negatively correlated with higher CO₂ levels. Adult numbers in single- and mixed-species tests were lower at sublethal CO₂ levels compared to ambient CO₂ levels at all the test temperatures and decreased in dry grain compared to damp grain. Although, a specific trend was not observed in population inhibition between mixed-species and single-species tests, overall the adult populations of T. castaneum were reduced in the presence of C. ferrugineus. A mathematical model was derived to predict the size of adult populations of T. castaneum alone, or in the presence of C. ferrugineus considering all the variables in this study. The model had an R² value of 0.72 but needs to be validated and refined with field data.     

Spatial distribution of some beetles infesting a feed mill with spatio-temporal dynamics of Oryzaephilus surinamensis, Tribolium castaneum and Tribolium confusum

Food-bait traps were used to study spatial and temporal distribution of the coleopteran fauna of the first floor of a feed mill in Central Italy. A total of 3396 beetles were captured during the 1-year survey. Beetles were most abundant during July and August and least abundant during February and March. Tribolium confusum du Val was the most abundant and widespread, followed by Oryzaephilus surinamensis (L.), Tribolium castaneum (Herbst), Attagenus brunneus Faldermann, Sitophilus oryzae (L.), and Stegobium paniceum (L.). The spatial patterns of annual catches of these six species were depicted by contour maps. In addition, high catches of O. surinamensis, T. castaneum and T. confusum, allowed construction of contour maps for monthly trap catches. The populations of A. brunneus were located in the bagging, milling, formulation and pelleting sites, and in the area around the conveyer belt. The highest populations of O. surinamensis were in the entry zone of the unloading pit and in the area around the conveyer belt with finished products. Sitophilus oryzae were present near the entry door of the unloading pit room, around the storage bins and in the storeroom. Populations of S. paniceum were found in the unloading pit as far as the conveyer belt and in a corner of storeroom. Tribolium castaneum was limited to a localized area in the room where raw material was processed and in the entry to the unloading pit room. The highest T. confusum populations were located in the milling, formulation and pelleting sites, and near balance and storage bins. Comparison of the spatio-temporal dynamics of these pests showed a segregation of populations, in both time and space, and a strong interaction among species is suggested. On the basis of our results, the spatial and temporal distributions are significantly affected by various factors, such as food availability, processing practices and temperature conditions in different areas.     

Survival and reproduction of Tribolium castaneum (Herbst), Rhyzopertha dominica (F.) and Sitophilus oryzae (L.) following periods of starvation

This study determined the starvation tolerance of Tribolium castaneum (Herbst), Rhyzopertha dominica (F.) and Sitophilus oryzae (L.) in terms of both adult survival and reproduction, the impact of starvation on reproduction not having been studied before. Experiments were conducted at 30 °C and 55% or 70% r.h. using a laboratory strain and a field strain of each species. The number of progeny was a better indicator of the impact of starvation on a species than adult survival. Tribolium castaneum was the most tolerant species, requiring up to 35 d starvation before no progeny were produced. Rhyzopertha dominica and S. oryzae required up to 8 d starvation before no progeny were produced. The results suggest that hygiene will have a greater impact on populations of S. oryzae and R. dominica than T. castaneum.     

Progress towards a multi-species lure: comparison of behavioural bioassay methods for multi-species attractants against three pests of stored grain

The object of the study reported here was to establish the best behavioural bioassay to ascertain the responses of three representative species of stored-grain insect pests to potentially attractive odours. The insects tested were Oryzaephilus surinamensis, Sitophilus granarius and Cryptolestes ferrugineus. Following consideration of different types of behavioural bioassay, the Y-tube olfactometer and the pitfall bioassay were chosen for comparison. Both were tested with a representative substance known to be attractive to the test species: carob (Ceratonia siliqua) aeration extract in pentane, and a pentane control. A number of physical and biological factors were taken into account in order to ascertain the most efficient bioassay method while retaining similarity to the natural environment of the insects. The Y-tube olfactometer was used with these species for the first time and it gave good differentiation between test and control with O. surinamensis and S. granarius but not with C. ferrugineus. With the pitfall bioassay, the optimum conditions for screening potential attractants for all three species were 20°C and 50% relative humidity, with a bioassay duration of 1 h. Therefore, the optimum bioassay method for mixed sex adults of the three species tested was the pitfall bioassay. The increased response to the attractant with increased temperature (from 15°C to 20°C) shows that changing temperature may affect the insects’ responses.     

Feasibility of protein fingerprinting technology for detecting Tribolium castaneum (Herbst) insect fragments in wheat flour

The red flour beetle, Tribolium castaneum (Herbst), is one of the predominant insects that cause serious decrease in quality of wheat flour. Protein mass fingerprinting has the potential to accurately detect insect parts in wheat flour, based on special peptides of T. castaneum. For determining the capability of protein fingerprinting for detecting insect parts in wheat flour, T. castaneum, wheat flour, and mixtures of T. castaneum and wheat flour were tested in this study. Three protein extraction buffers: 50% acetonitrile; sterilized water; and the buffer of 10 mM imidazole pH 7.0, 1 mM phenylthiourea, 1% Triton X-100 and protease inhibitor cocktail (PTU buffer), were compared for insect protein detection. An Agilent 1100 reversed phase high performance liquid chromatography (RP-HPLC) instrument was used for peptide purification and fractionation. Matrix-assisted laser desorption/ionization (MALDI) was used to read the RP-HPLC fractions, and the results were analyzed using sMART Engine, details given later. Fifty percent acetonitrile was adopted as protein extract buffer. Since very limited information about T. castaneum protein can be found in the database of sMART and National Center for Biotechnology Information (NCBI), peptide mapping method was used for identifying insect from the wheat flour. Peptide mapping method, which could set up biomarkers for each type of insect, was proven to be a more effective and practical way to detect insect contamination. This peptide mapping technique could obtain 78 biomarkers of T. castaneum, when T. castaneum was 0.1% in wheat flour by mass. This sensitivity needs to be increased further during future research.