Venom gland components of the ectoparasitoid wasp, Anisopteromalus calandrae.

The wasp Anisopteromalus calandrae is a small ectoparasitoid that attacks stored product pest beetle larvae that develop inside grain kernels, and is thus a potential insect control tool. The components of A. calandrae venom have not been studied, but venom from other organisms contains proteins with potential applications, such as pest management tools and treatments for human diseases. We dissected female A. calandrae and collected venom and associated glands. Using high throughput sequencing, a venom gland transcriptome was assembled that contained 45,432 contigs, 25,726 of which had BLASTx hits. The majority of hits were to Nasonia vitripennis, an ectoparasitoid from the same taxonomic family, as well as other bees, wasps, and ants. Gene ontology grouped sequences into eleven molecular functions, among which binding and catalytic activity had the most representatives. In this study, we highlighted the most abundant sequences, including those that are likely the functional components of the venom. Specifically, we focused on genes encoding proteins potentially involved in host developmental arrest, disrupting the host immune system, host paralysis, and transcripts that support these functions. Our report is the first to characterize components of the A. calandrae venom gland that may be useful as control tools for insect pests and other applications.

Sampling stored-product insect pests: a comparison of four statistical sampling models for probability of pest detection.

BACKGROUND: Developing sampling strategies to target biological pests such as insects in stored grain is inherently difficult owing to species biology and behavioural characteristics. The design of robust sampling programmes should be based on an underlying statistical distribution that is sufficiently flexible to capture variations in the spatial distribution of the target species. RESULTS: Comparisons are made of the accuracy of four probability-of-detection sampling models - the negative binomial model,(1) the Poisson model,(1) the double logarithmic model(2) and the compound model(3) - for detection of insects over a broad range of insect densities. Although the double log and negative binomial models performed well under specific conditions, it is shown that, of the four models examined, the compound model performed the best over a broad range of insect spatial distributions and densities. In particular, this model predicted well the number of samples required when insect density was high and clumped within experimental storages. CONCLUSIONS: This paper reinforces the need for effective sampling programs designed to detect insects over a broad range of spatial distributions. The compound model is robust over a broad range of insect densities and leads to substantial improvement in detection probabilities within highly variable systems such as grain storage.

Impact of varying levels of sanitation on mortality of Tribolium castaneum eggs and adults during heat treatment of a pilot flour mill.

The influence of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), an economically important pest in flour mills, was investigated in a pilot flour mill subjected to two, 24-h heat treatments. One hundred eggs or 100 adults of T. castaneum were exposed inside each 20-cm diameter by 15-cm high PVC rings holding 0.1-, 0.2-, 1.0-, 3.0-, 6.0-, and 10.0-cm-deep wheat flour to simulate different sanitation levels that may exist in a flour mill. These rings were placed on the first and third floors of a pilot flour mill. On the first floor, temperatures inside rings with eggs reached 50 degrees C in 7-11 h only in 0.1- and 0.2-cm-deep flour treatments. In all other treatments the maximum temperatures attained generally were below 50 degrees C and inversely related to flour depth. Adults of T. castaneum on this floor were less susceptible than eggs. The egg mortality decreased linearly with an increase in flour depth, whereas that of adults decreased exponentially. All eggs and adults in rings on the third floor were killed irrespective of flour depth, because temperatures inside rings reached 50 degrees C in 15-17 h and were held above 50 degrees C for 6-8 h with the maximum temperatures ranging between 55.0 and 57.0 degrees C. Although the protective effects of flour on survival of T. castaneum eggs and adults were evident only if temperatures did not reach 50 degrees C, removal of flour accumulations is essential to improve heat treatment effectiveness.

Susceptibility of Tribolium castaneum life stages exposed to elevated temperatures during heat treatments of a pilot flour mill: influence of sanitation, temperatures attained among mills floors, and costs.

The influence of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), was investigated in a pilot flour mill subjected to three, 24-h heat treatments by using forced-air gas heaters fueled by propane. Two sanitation levels, dusting of wheat flour and 2-cm-deep flour, were created in 25 plastic bioassay boxes, each holding eggs, young larvae, old larvae, pupae, and adults of T. castaneum plus two temperature sensors. Data loggers (48) were placed on the five mill floors to record air temperatures. The time required to reach 50 degrees C, time above 50 degrees C, and the maximum temperature among mill floors and in bioassay boxes were measured. The maximum temperature in bioassay boxes and in the mill was lower on the first floor than on other floors. This trend was apparent in time required to reach 50 degrees C and time above 50 degrees C, especially in compartments with 2-cm-deep flour. The mean +/- SE mortality of T. castaneum life stages on the first floor was 55.5 +/- 12.9-98.6 +/- 0.8%; it was 93.2 +/- 6.7-100 +/- 0.0% on other floors. Adults were the least susceptible stage. Mortality of T. castaneum stages in compartments with 2-cm-deep flour was generally lower than those with flour dust. Costs for the three heat treatments ranged from US$27,438 to $28,838. An effective heat treatment can be conducted within 24 h, provided temperatures on mill floors reach 50 degrees C in 8-12 h and are held above 50 degrees C for at least 10-14 h, with maximum temperatures held between 50 and 60 degrees C.

Effects of flour conditioning on cannibalism of T. castaneum eggs and pupae.

Cannibalism is a very important factor regulating population dynamics of the red flour beetle. After several days of feeding, the flour becomes conditioned by the beetles, which can affect rates of cannibalism. Flour conditioning is caused by an accumulation of feces, pheromones, and ethylquinone, which is a repellent produced by the beetles. We determined the effect of five different levels of flour conditioning on cannibalism of red flour beetle eggs and pupae by adult and larval stages. Larvae had the highest rates of egg cannibalism (12 eggs eaten over the 4-d period) followed by female adults (seven eggs consumed). Adult males had the lowest rates of cannibalism with only two eggs consumed. Cannibalism of eggs by females was correlated negatively with the level of flour conditioning. There was no effect of flour conditioning on egg or pupal cannibalism by larvae or adult males. Cannibalism by adult females may decrease as the level of flour conditioning increases because females may spend less time tunneling in highly conditioned flour and more time trying to disperse to other areas that are better for oviposition.

Susceptibility of Lasioderma serricorne (Coleoptera: Anobiidae) life stages to elevated temperatures used during structural heat treatments.

Heat treatment of food-processing facilities involves using elevated temperatures (50-60 degrees C for 24-36 h) for management of stored-product insects. Heat treatment is a viable alternative to the fumigant methyl bromide, which is phased out in the United States as of 2005 because of its adverse effects on the stratospheric ozone. Very little is known about responses of the cigarette beetle, Lasioderma serricorne (F.) (Coleoptera: Anobiidae), a pest associated with food-processing facilities, to elevated temperatures. Responses of L. serricorne life stages to elevated temperatures were evaluated to identify the most heat-tolerant stage. Exposure of eggs, young larvae, old larvae, and adults during heat treatment of a food-processing facility did not clearly show a life stage to be heat tolerant. In the laboratory, exposure of eggs, young larvae, old larvae, pupae, and adults at fixed times to 46, 50, and 54 degrees C and 22% RH indicated eggs to be the most heat-tolerant stage. Time-mortality responses at each of these three temperatures showed that the time for 99% mortality (LT99) based on egg hatchability and egg-to-adult emergence was not significantly different from one another at each temperature. Egg hatchability alone can be used to determine susceptibility to elevated temperatures between 46 and 54 degrees C. The LT99 based on egg hatchability and egg-to-adult emergence at 46 degrees C was 605 and 598 min, respectively, and it decreased to 190 and 166 min at 50 degrees C and 39 and 38 min at 54 degrees C. An exponential decay equation best described LT99 as a function of temperature for pooled data based on egg hatchability and egg-to-adult emergence. Our results suggest that during structural heat treatments eggs should be used in bioassays for gauging heat treatment effectiveness, because treatments aimed at controlling eggs should be able to control all other L. serricorne life stages.

Susceptibility of Tribolium castaneum (Coleoptera: Tenebrionidae) life stages to flameless catalytic infrared radiation.

The susceptibility of various life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), a pest of stored wheat, Triticum aestivum L., to flameless catalytic infrared radiation in the 3-7-microm range was evaluated in the laboratory. Immature stages were collected from flour infested with T. castaneum adults only for 1 d. Stages collected after 1 d represented eggs (collected on day 0); those collected after 7, 14, and 21 d from day 0 represented larvae in different developmental stages, whereas those collected after 24 d represented pupae. Adults (2 wk old) were collected after 42 d. Each of these stages was exposed for 45 or 60 s in 113.5 or 227.0 g of wheat at a distance of 8.0 or 12.7 cm from a bench top infrared emitter. The mean temperatures attained during exposures were measured continuously using a noncontact infrared thermometer connected to a computer. The mean grain temperatures attained increased with an increase in exposure time and were inversely related to distance from the emitter. Grain quantity least influenced mean temperatures attained. Pupae were the least susceptible stage and larvae collected after 7 d were the most susceptible stage. Variation in probability of death of various life stages decreased with an increase in mean grain temperatures attained. All life stages were killed after a 60-s exposure at a distance of 8.0 cm from the emitter in 113.5 g of wheat, where the mean +/- SE temperatures attained ranged from 107.6 +/- 1.2 to 111.4 +/- 0.5 degrees C. Our laboratory results using small grain quantities and short exposure times showed that flameless catalytic infrared radiation can be a valuable tool for managing insects in stored organic and nonorganic wheat.

Susceptibility of various life stages of Rhyzopertha dominica (Coleoptera: Bostrichidae) to flameless catalytic infrared radiation.

In laboratory experiments, a flameless catalytic infrared emitter, fueled by propane, was used to disinfest hard red winter wheat, Triticum aestivum L., containing different life stages of the lesser grain borer, Rhyzopertha dominica (F.), an economically important insect species associated with stored wheat in Kansas. The emitter generates infrared radiation in the 3-7-microm range. The life stages of R. dominica exposed to infrared radiation included eggs, larvae in different stages of development, pupae, and 2-wk-old adults. A noncontact infrared thermometer measured grain temperatures continuously during exposures of infested wheat to infrared radiation. The grain temperatures attained were influenced by wheat quantity; distance from the emitter; and exposure time, which in turn influenced effectiveness against various life stages of H. dominica. In general, higher grain temperatures were attained in 113.5 g of wheat as opposed to 227.0 g, and at 8.0 cm from the emitter surface rather than at 12.7 cm, and during a 60-s exposure compared with a 45-s exposure. Logistic regression indicated the probability of death of various life stages of R. dominica was temperature dependent. The log odds ratios showed old larvae were less susceptible to infrared radiation than young larvae. Approximately > or = 94% mortality of all R. dominica life stages occurred when using 113.5 g of wheat, exposed for 60 s at a distance of 8.0 cm from the emitter, resulting in mean +/- SE wheat temperatures that ranged between 107.6 +/- 1.4 and 113.5 +/- 0.5 degrees C. Our results with small grain quantities show flameless catalytic infrared technology to be a promising tool for disinfestation of stored wheat.

Effects of intra- and interpatch host density on egg parasitism by three species of Trichogramma.

Host-foraging responses to different intra- and interpatch densities were used to assess three Trichogramma spp. (Hymenoptera: Trichogrammatidae) Trichogramma deion Pinto and Oatman, T. ostriniae Pang and Chen, and T. pretiosum Riley - as potential biological control agents for the Indian meal moth, Plodia interpunctella Hübner (Lepidoptera: Pyralidae). Single naïve females were allowed 6 h to forage in Plexiglas arenas with four different spatial arrangements of host eggs, nine single-egg patches), nine four-egg patches, 36 single-egg patches, and 36 four-egg patches. No significant differences were found among species in the number of patches parasitized. As expected, all three species parasitized the most eggs in the 36 four-egg patch treatment and the least in the nine single-egg patch treatment. T. deion parasitized significantly more eggs than T. pretiosum on the nine four-egg patches. T. ostriniae parasitized significantly more patches when intrapatch density was greater, regardless of interpatch density. In contrast, T. deion only parasitized more patches at the greater intrapatch density when the interpatch density was low. Patch density had no effect on T. pretiosum. The spatial pattern of parasitism was more aggregated for T. deion and T. ostriniae in the 36 four-egg patches treatment compared to the 36 single-egg patches treatment. Therefore, intrapatch density was more important than interpatch density for T. ostriniae, and potentially for T. deion, but not for T. pretiosum. T. deion may be the best candidate for augmentative biological control because it parasitized either slightly or significantly more eggs than the other two species in all four treatments. Furthermore, the pattern of parasitism by T. deion in the 36 four-egg patches treatment was the most aggregated among the three species, suggesting a more thorough searching pattern. In contrast, T. pretiosum had the least aggregated pattern of parasitism and therefore may have used a more random foraging pattern.

Condições higiênico-sanitárias da pimenta do reino em pó (Piper nigrum L) com o emprego de duas diferentes técnicas para detecção de sujidades leves / Assessment of hygienic conditions of ground pepper (Piper nigrum L) by means of two methodologies for detecting the light filth

A pimenta do reino deve ser colhida, processada, embalada em condições ideais para evitar a presença de matérias estranhas. Com o objetivo de avaliar as condições higiênicas da pimenta do reino em pó, comercializada em São Paulo, foram analisadas 22 amostras por meio de duas diferentes técnicas no período de maio a setembro de 2006. Para a análise de sujidades leves foi utilizada a técnica de flutuação segundo AOAC e ensaio imunoenzimático – ELISA. Observou-se que 100% das amostras continham fragmentos de insetos e em várias amostras foram demonstrados mais de um tipo de matéria estranha; e 23% das amostras mostraram ser impróprias ao consumo em virtude da presença de pelos de roedor. Para realizar o ELISA, foram adicionados os padrões constituídos de 1, 2, 4, 8 e 10 insetos para estimar o número de insetos nas amostras em análise, por meio de valores de densidade óptica (DO). Verificou-se que 36,4% das amostras apresentaram valores de DO próximos aos correspondentes ao do padrão 8 insetos, 40,9% ao do padrão 4 insetos, 18,2% ao do padrão 10 e 4,5% ao do padrão 2 insetos. A técnica descrita na AOAC mostra ser mais apropriada, em função de sua eficiência de detecção de insetos, bem como de outras matérias estranhas, enquanto que o ELISA detecta especificamente a miosina presente no músculo dos insetos, cuja degradação ocorre com o passar do tempo.

Host-foraging success of three species of Trichogramma (Hymenoptera: Trichogrammatidae) in a simulated retail environment.

Three species of trichogrammatid egg parasitoids (Trichogramma deion Pinto & Oatman, Trichogramma ostriniae Pang & Chen, and Trichogramma pretiosum Riley) (Hymenoptera: Trichogrammatidae) were evaluated under laboratory conditions as potential biological control agents for the Indianmeal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), on retail shelves. A single shelving unit was used in each trial and a grid of sentinel egg disks was used to evaluate foraging success. The shelving consisted of pallet units with five shelves that were either bare or stocked with empty cereal boxes. In each replicate, approximately 500 female Trichogramma were released at the center of the shelving unit and allowed to forage for 48 h. Percentage of egg parasitism and percentage of host egg mortality were recorded after 7 d. Foraging success as well as the spatial pattern of parasitism differed significantly among the three Trichogramma species. Percentage of egg parasitism was approximately 4 times greater for T. deion than for T. ostriniae or T. pretiosum. The vertical distribution of parasitism by T. deion was also more uniform than for the other two species. In addition, the presence of packaging affected the foraging efficiency of T. ostriniae and T. pretiosum but not T. deion. Based on these findings, Trichogramma deion may be the best-suited candidate for augmentative biological control of P. interpunctella in retail stores, and a central release point of T. deion will likely provide adequate coverage of products on pallet-type shelving.

Biological control of Indianmeal moth (Lepidoptera: Pyralidae) on finished stored products using egg and larval parasitoids.

Biological control using hymenopteran parasitoids presents an attractive alternative to insecticides for reducing infestations and damage from the Indianmeal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), in retail and warehouse environments. We examined the potential for using combinations of the egg parasitoid Trichogramma deion Riley (Hymenoptera: Trichogrammatidae), and the larval parasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae) for preventing infestations of P. interpunctella in coarse-ground cornmeal as well as the influence of packaging on parasitoid effectiveness. Treatments included one or both parasitoids and either cornmeal infested with P. interpunctella eggs or eggs deposited on the surface of plastic bags containing cornmeal. H. hebetor had a significant impact on the number of live P. interpunctella, suppressing populations by approximately 71% in both unbagged and bagged cornmeal. In contrast, T. deion did not suppress P. interpunctella in unbagged cornmeal. However, when released on bagged cornmeal, T. deion significantly increased the level of pest suppression (87%) over bagging alone (15%). When H. hebetor was added to bagged cornmeal, there was a significant reduction of live P. interpunctella compared with the control (70.6%), with a further reduction observed when T. deion was added (96.7%). These findings suggest that, in most situations, a combined release of both T. deion and H. hebetor would have the greatest impact; because even though packaging may protect most of the stored products, there are usually areas in the storage landscape where poor sanitation is present.

Effects of shelf architecture and parasitoid release height on biological control of Plodia interpunctella (Lepidoptera: Pyralidae) eggs by Trichogramma deion (Hymenoptera: Trichogrammatidae).

The effects of shelving type, packaging, and release height on success of Trichogramma deion Pinto & Oatman (Hymenoptera: Trichogrammatidae) parasitizing Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) eggs was studied under laboratory conditions. In trials on multipletiered gondola-type or open shelving units, with or without packaging, foraging success was evaluated by comparing parasitism and total mortality rates of sentinel egg disks among shelves after a single point-release of T. deion. Results showed that T. deion parasitized more egg disks and killed more total eggs on open shelves than on gondola shelving. The presence of packaging had no effect on parasitoid foraging on open shelves; however, packaging did interfere with parasitism of P. interpunctella eggs on gondola shelving. Egg parasitism and mortality patterns among shelves were not as evenly distributed on gondola-type shelving compared with open shelving. On gondola shelves without packages, changing the release point of T. deion from the middle to the lowest shelf shifted the distribution of parasitism toward the floor. Gondola shelving, especially in the presence of packaging, reduced foraging efficiency of T. deion for P. interpunctella eggs. Thus, to attain adequate control of P. interpunctella, it may be necessary to use two release heights on gondola shelving.

Reproductive performance of Tribolium castaneum (Coleoptera: Tenebrionidae) exposed to the minimum heat treatment temperature as pupae and adults.

Managing stored-product insect pests by heating the ambient air of a food-processing facility to high temperatures (50-60 degrees C), also referred to as heat treatment, is an effective technology that has been used since the early 1900s. The minimum temperature during heat treatment for effective disinfestation is 50 degrees C. The effect of sublethal exposures to 50 degrees C on the reproductive performance of stored-product insects associated with food-processing facilities is unknown. The red flour beetle, Tribolium castaneum (Herbst), is a pest commonly found in food-processing facilities worldwide. The adverse effects on fecundity, egg-to-adult survival, and progeny production of T. castaneum exposed as 1-d-old pupae and 2-wk-old adults to 50 degrees C for 60 and 39 min, respectively, were determined in the laboratory. Pupae and adults exposed for the same time periods at 28 degrees C served as the control treatment. Four possible reciprocal crosses were carried out among adults from the heat-treated (50 degrees C) and control (28 degrees C) treatments. The number of eggs produced during the first 2 wk of adult life, survival of these eggs to adulthood, and adult progeny production after 2 and 8 wk of oviposition in treatments representing all four reciprocal crosses were determined. Fecundity, egg-to-adult survival, and adult progeny production decreased by 17-63, 52-63, and 66-78%, respectively, when males, females, and both males and females were exposed to 50 degrees C. These effects were relatively more pronounced in treatments in which pupae were exposed to the high temperature compared to adults, and in exposed females than in males. The impaired reproductive performance in T. castaneum pupae and adults surviving sublethal exposures to the minimum heat treatment temperature is valuable for understanding population rebound following a heat treatment intervention.

Detection of stored-grain insect infestation in wheat transported in railroad hopper-cars.

Levels of insect infestation, insect spatial distribution, and the relationship between the number of insect-damaged kernels (IDK) and the number of insects present in grain samples in three-hopper railcars transporting wheat from country elevators to a mill were studied. Six of eight sampled railcars were infested with more than two species of insects. The most abundant species collected were the lesser grain borer, Rhyzopertha dominica (F.), and rusty grain beetle, Cryptolestes ferrugineus (Stephens), with the larval stage of the two species being the most prevalent (>90%). The spatial distributions of these two species within the grain mass were typically clumped in railcar compartments containing >0.4 insect/2.75-kg sample of wheat, and these foci of high-infestation levels varied in compartments within the railcars and among the sampled railcars. There were no significant correlations between IDK and insect density for any of the different stage-specific insect populations that were collected in the grain samples. Mean numbers of immatures and IDK differed among railcars and compartments within railcars, but not among grain depths. Number of insects in the first discharge sample was not correlated with mean numbers of insects in the entire compartment. This indicates that each compartment of a railcar should be sampled to determine level of insect infestation but that sampling at different depths within a compartment is less important.

United States Department of Agriculture-Agricultural Research Service stored-grain areawide integrated pest management program.

The USDA Agricultural Research Service (ARS) funded a demonstration project (1998-2002) for areawide IPM for stored wheat in Kansas and Oklahoma. This project was a collaboration of researchers at the ARS Grain Marketing and Production Research Center in Manhattan, Kansas, Kansas State University, and Oklahoma State University. The project utilized two elevator networks, one in each state, for a total of 28 grain elevators. These elevators stored approximately 31 million bushels of wheat, which is approximately 1.2% of the annual national production. Stored wheat was followed as it moved from farm to the country elevator and finally to the terminal elevator. During this study, thousands of grain samples were taken in concrete elevator silos. Wheat stored at elevators was frequently infested by several insect species, which sometimes reached high numbers and damaged the grain. Fumigation using aluminum phosphide pellets was the main method for managing these insect pests in elevators in the USA. Fumigation decisions tended to be based on past experience with controlling stored-grain insects, or were calendar based. Integrated pest management (IPM) requires sampling and risk benefit analysis. We found that the best sampling method for estimating insect density, without turning the grain from one bin to another, was the vacuum probe sampler. Decision support software, Stored Grain Advisor Pro (SGA Pro) was developed that interprets insect sampling data, and provides grain managers with a risk analysis report detailing which bins are at low, moderate or high risk for insect-caused economic losses. Insect density was predicted up to three months in the future based on current insect density, grain temperature and moisture. Because sampling costs money, there is a trade-off between frequency of sampling and the cost of fumigation. The insect growth model in SGA Pro reduces the need to sample as often, thereby making the program more cost-effective. SGA Pro was validated during the final year of the areawide program. Based on data from 533 bins, SGA Pro accurately predicted which bins were at low, moderate or high risk. Only in two out of 533 bins did SGA Pro incorrectly predict bins as being low risk and, in both cases, insect density was only high (> two insects kg(-1)) at the surface, which suggested recent immigration. SGA Pro is superior to calendar-based management because it ensures that grain is only treated when insect densities exceed economic thresholds (two insects kg(-1)). This approach will reduce the frequency of fumigation while maintaining high grain quality. Minimizing the use of fumigant improves worker safety and reduces both control costs and harm to the environment.