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.     

Simulation model of Rhyzopertha dominica population dynamics in concrete grain bins

Rhyzopertha dominica is one of the most damaging insect pests in grain elevators and causes millions of dollars worth of stored grain losses annually in the USA. A simulation model was developed for predicting R. dominica population dynamics in concrete grain bins. The model used a two-dimensional representation of a cylindrical concrete bin (33 m tall×6.4 m wide), and used hourly weather data to predict changes in grain temperature. Output from the grain bin temperature and moisture module was used by the insect module to predict changes in insect density in 32 different bin regions. When compared to validation data from nine grain bins, the model accurately predicted insect vertical distribution and insect density. In December, the highest insect density was in the top center of the grain mass, and decreased steadily with increasing depth and towards the periphery of the grain mass. R. dominica attains this spatial distribution because immigration is primarily through the top of the bin, and higher populations occur in the interior of the grain mass because of warmer temperatures there. Initially, the model underestimated actual insect density in the grain bins. We increased the immigration rate by 50% and this resulted in a much better prediction of R. dominica density by the model. From 20 September to 14 December, populations of R. dominica increased from 0.1 to 3.5 insects per kg of wheat.     

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.     

Moisture content gradient and ventilation in stored wheat affect movement and distribution of Oryzaephilus surinamensis and have implications for pest monitoring

This study was carried out primarily to ascertain whether the movement of Oryzaephilus surinamensis and Sitophilus granarius from low to high humidity zones occurs in bins of wheat and whether aeration of the grain (10 m³/h/t) affects this movement. The second aim was to ascertain the best placement of insect detection traps under the different conditions. Insects were introduced into the lower half of the grain in the bins and their movement was monitored using traps placed at various depths in the grain. Sitophilus granarius did not move through the grain into the top layer regardless of the moisture content, temperature or aeration status of the grain. More O. surinamensis were caught in unventilated bins than in ventilated bins. More insects were caught in the ventilated bins containing layers of both dry and wet grain than in the bins containing only dry grain. The spatio-temporal distribution of O. surinamensis varied significantly. The depth at which insects were trapped varied between treatments: in ventilated dry grain, most insects were trapped at the surface; in ventilated wet and dry grain, most insects were trapped at 10 cm and 0.75 m; in unventilated wet and dry grain, the vast majority of the insects were trapped at 0.75 m. Very few insects were trapped at 1.75 m regardless of the treatment. The proportions of the initial population of O. surinamensis which were recaptured in the top layer of grain varied between treatments. Most were recaptured in the unventilated bins containing wet and dry grain followed by ventilated bins containing wet and dry grain. The smallest proportion of the population was recaptured in the ventilated bins containing only dry grain. Immediate practical implications for pest monitoring based on physical control measures in use are discussed.     

Heat treatment for disinfestation of empty grain storage bins

An alternative to fumigants and insecticides for controlling stored-product insects in empty grain storage bins prior to filling is heat treatment, in which the temperature is quickly raised to a minimum of 50 °C and held there for 2-4 h. Effectiveness of heat treatment on empty grain storage bins was evaluated for five commercial propane and electric heat-treatment systems by measuring air temperature and associated mortality of Tribolium castaneum (Herbst), the red flour beetle, Sitophilus oryzae (L.), the rice weevil, and Rhyzopertha dominica (F.), the lesser grain borer, exposed for different time intervals. Eleven locations, six above and five below the drying floor, were monitored for air temperature and associated mortality of the three insect species, using arenas initially stocked with live adult insects. Data were analyzed separately for each heating system, with floor location and time interval as main effects for insect mortality. A high-output propane heater (29 kW) produced 100% mortality in 2 h for the three insect species at all test locations. An electric duct-heater system (18 kW) also produced 100% mortality at all test locations after 40 h when aided by a complicated interior heat-distribution system. The other three systems produced less than 100% mortality.     

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 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.     

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.     

Impact of resistance on the efficacy of binary combinations of spinosad, chlorpyrifos-methyl and s-methoprene against five stored-grain beetles

Laboratory experiments were conducted to determine the efficacy of spinosad (a biopesticide), chlorpyrifos-methyl (an organophosphorus compound (OP)) and s-methoprene (a juvenile hormone analogue) applied alone and in binary combinations against five stored-grain beetles in wheat. There were three strains of Rhyzopertha dominica, and one strain each of Sitophilus oryzae, Tribolium castaneum, Oryzaephilus surinamensis and Cryptolestes ferrugineus. These strains were chosen to represent a range of possible resistant genotypes, exhibiting resistance to organophosphates, pyrethroids or methoprene. Treatments were applied at rates that are registered or likely to be registered in Australia. Adults were exposed to freshly treated wheat for 2 weeks, and the effects of treatments on mortality and reproduction were determined. No single protectant or protectant combination controlled all insect strains, based on the criterion of >99% reduction in the number of live F₁ adults relative to the control. The most effective combinations were spinosad at 1 mg kg⁻¹+chlorpyrifos-methyl at 10 mg kg⁻¹ which controlled all strains except for OP-resistant O. surinamensis, and chlorpyrifos-methyl at 10 mg kg⁻¹+s-methoprene at 0.6 mg kg⁻¹ which controlled all strains except for methoprene-resistant R. dominica. The results of this study demonstrate the difficulty in Australia, and potentially other countries which use protectants, of finding protectant treatments to control a broad range of pest species in the face of resistance development.     

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.     

Movement of Cryptolestes ferrugineus (Coleoptera: Cucujidae) in grain columns containing pockets of high moisture content wheat and carbon dioxide gradients

The movement of the adult rusty grain beetle, Cryptolestes ferrugineus (Stephens), in 100 cm long, 10 cm diameter columns, filled with wheat of uniform or non-uniform moisture content (m.c.), was determined for horizontal and vertical placement of columns at 30°C.In horizontal columns filled with wheat of 13.5% uniform m.c., adult C. ferrugineus exhibited a trend of distributing themselves uniformly in the columns after 5 d. In the vertical columns filled with 14.7% uniform m.c. wheat, 44% of the insects moved downward after 7 d showing positive geotaxism. In the horizontal or vertical columns containing pockets of high m.c. wheat, 43-48% of the beetles aggregated in the region of high m.c. after 7 d exhibiting a hygrotactic response. Under CO₂ gradients in the columns filled with uniform m.c. wheat, 65% of C. ferrugineus moved towards higher levels of CO₂ after 1 and 5 d. Also, high levels of CO₂ were more influential in attracting C. ferrugineus (60%) after 1 d in horizontal columns containing high m.c. wheat at one end and high CO₂ levels at the other. In the vertical columns filled with pockets of high m.c. wheat and subjected to CO₂ gradients, movement of C. ferrugineus was more influenced by the combined effects of any two factors out of the three, viz., gravity, higher levels of CO₂ and high m.c. than a single factor.     

Mortality and movement of Cryptolestes ferrugineus and Rhyzopertha dominica in response to cooling in 300-kg grain bulks

We examined the effect of low temperatures on insect population density and mortality of adult and immature stages of Cryptolestes ferrugineus and Rhyzopertha dominica in three barrels holding 300 kg of wheat each. Barrels were infested with 2 insects/species/kg and left to develop at 30 °C for 5 months. Temperature of the grain mass before cooling was 38 °C in the centre to a low of 28 °C on the periphery. On 11 February the building door was opened to allow for cooling of the grain. During the 3 weeks of cooling, temperatures in the centre reached as low as −5 °C. Segregated (11 segments) grain trier samples were collected from three locations in each barrel, from February until March 2013 (6 dates), sieved, and the number of live and dead adults counted. Sieved grain was held for 5 weeks 30 °C and emerged adults counted to give an estimation of immature stages within the grain at the time of sampling. Rhyzopertha dominica adults were found mainly in the top 40 cm of the barrel, and there was little movement to the centre as the grain mass cooled. The initial distribution of C. ferrugineus adults was more complex; in two barrels adults were mostly in the centre of the grain mass, while in the other barrel, more adults were found on the periphery. When the grain mass cooled, C. ferrugineus adults in all barrels were found mostly in the centre. On day 0, the average number of live adults of C. ferrugineus and R. dominica were 232 ± 56 and 414 ± 48 insects/100 g, respectively. Rhyzopertha dominica adults were more susceptible to low temperatures than C. ferrugineus. The density of immatures of C. ferrugineus and R. dominica were similar at 0 d, there was a decline over the 21 d, but there was some survival at day 21.     

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.     

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.     

Toxicity of ethyl formate to adult Sitophilus oryzae (L.), Tribolium castaneum (herbst) and Rhyzopertha dominica (F.)

Fumigations were conducted using a continuous flow-through laboratory process to maintain constant concentrations of ethyl formate and low levels (<0.8%) of respiratory carbon dioxide. The procedure minimised the effects of sorption by exposing test insects without media and minimised the effect of carbon dioxide by use of continuous flow. The concentration×time (Ct) products of ethyl formate for adult Sitophilus oryzae, Tribolium castaneum and Rhyzopertha dominica at 25 °C and 70% relative humidity for the 6 h exposure were, respectively: (1) LD₅₀ 107.8, 108.8 and 72.8 mg h L⁻¹ and (2) LD_99.5 207.4, 167.1 and 122.2 mg h L⁻¹. Endpoint mortality was reached within 24 h of initial exposure.     

Enhanced fumigant toxicity of allyl acetate to stored-product beetles in the presence of carbon dioxide

Toxicity of allyl acetate (5-25 mg/l doses) to mixed-age cultures of stored-product beetles including Cryptolestes ferrugineus, Lasioderma serricorne, Oryzaephilus surinamensis, Rhyzopertha dominica, Sitophilus oryzae and Tribolium castaneum in the presence of carbon dioxide (CO₂) (10% and 20%) with a 48-h exposure period was studied in the laboratory at 27±2 °C. Depending on dosage and the insect species, the fumigant toxicity of allyl acetate was enhanced by CO₂. At most of the allyl acetate+CO₂ combinations, increased mortality was observed in C. ferrugineus, O. surinamensis and S. oryzae. In L. serricorne and T. castaneum, which are tolerant to allyl acetate, higher mortality due to CO₂ was achieved at selected dose combinations only (e.g. 15 mg/l allyl acetate+20% CO₂). Significant increase in mortality of R. dominica (the most susceptible species to allyl acetate) exposed to allyl acetate in the presence of CO₂ was not evident except at the lower dose of 5 mg/l allyl acetate+CO₂ that caused 31.7% mortality. The mortality data show that CO₂ could be used as an adjuvant for allyl acetate.     

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.     

Effect of low pressures on the survival of cocoa pests at 18°C

This study forms part of an effort to eliminate the need for fumigation with methyl bromide to control insect infestations in stored cocoa beans, through development of novel alternative vacuum-hermetic technology. In this communication, the effects of low pressures and exposure time were studied on the mortality of insects at a temperature of 18°C, chosen to simulate cocoa bean storage conditions in temperate climates.Three insect species were used, two of which are major pests of cocoa beans in producer countries, Ephestia cautella (Walker), and Tribolium castaneum (Herbst), while the third, Oryzaephilus surinamensis (L.), is a potential storage pest in temperate climates. For T. castaneum and E. cautella the egg stage was the most resistant to 55±10 mm Hg at 18°C, the times needed to obtain 99% egg mortality were 96 and 149 h, respectively. For O. surinamensis, the adult stage was the most resistant with 164 h being required to obtain 99% mortality.     

Detection of Rhyzopertha dominica larvae in stored wheat using ELISA: The impact of myosin degradation following fumigation

Hard red winter wheat kernels were infested with eggs of Rhyzopertha dominica. After 20 d, when the larvae reached the fourth instar, they were killed by exposing the infested kernels to phosphine gas for 24 h. The infested kernels were then divided into four portions and treated as follows: one portion was immediately frozen at −80 °C to avoid myosin degradation; the other three portions were kept at 32 °C and 65% relative humidity, and then frozen at −80 °C after 14, 28, and 56 d post-fumigation, respectively. Each treatment was replicated five times. Myosin was measured using a commercial enzyme linked immunosorbent assay (ELISA) method that specifically detects this protein (Biotect^®, Austin, TX). Myosin degradation was most rapid in the first 2 weeks after the larvae were killed, decreasing from 1.672 to 0.695 ng/well during this period (a 58.4% reduction). There were no significant differences in myosin degradation between samples that were 14, 28, and 56 d post-fumigation. Grain is often fumigated to control insects. Frequently, this occurs many weeks before the grain is milled and may be repeated during the storage period. Therefore, estimates using the ELISA test may underestimate internal insect infestation because of myosin degradation. Insect fragment estimates for previously fumigated grain could be underestimated by as much as 58%.     

Toxicity of coumaran to stored products beetles

The fumigant activity of coumaran was tested against mixed-age cultures of the rice weevil (Sitophilus oryzae L.), the lesser grain borer (Rhyzopertha dominica F.), the rust-red flour beetle (Tribolium castaneum Herbst), saw-toothed grain beetle (Oryzaephilus surinamensis L.) and flat grain beetle (Cryptolestes ferrugineus Stephens). T. castaneum was tolerant to coumaran, for total population extinction was achieved in 72 h exposure at 50 μg/L dose only. In contrast all developmental stages of O. surinamensis, C. ferrugineus and R. dominica were quite susceptible 100% mortality was attained at < 50 μg/L doses in 24 or 72 h exposure. Further results showed that T. castaneum, generally tolerant pest to fumigants and R. dominica, a predominant stored grain pest in India, could be controlled by coumaran. The findings indicate that, coumaran is good source for alternate to methylbromide for effective control of stored grain insect pests.