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.     

Influence of environmental and physical factors on capture of Tribolium castaneum (Coleoptera: Tenebrionidae) in a flour mill

Variation in environmental and physical factors within food processing facilities can influence both the distribution of stored-product pests and trapping efficiency. Data from a long-term Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) monitoring program was used to evaluate spatial variation in captures among trap locations and to determine relationships with environmental and physical variables. From the complete monitoring data set, different subsets were created for the cool and warm seasons, and period of time when environmental and physical factors were measured (2009-2010), with all data sets showing significant differences among trap locations in terms of beetle captures and proportion of time that traps exceeded 2.5 beetles per trap per monitoring period. There was also considerable temporal variation in distribution among the different levels of the mill. Among the environmental and physical variables measured, mean temperature and flour dust accumulation showed the most significant positive relationships with variation of beetle captures at trap locations. More beetles were captured in traps located in close proximity to milling equipment. Presence of equipment near traps was also associated with an increase in flour dust accumulation and temperature. Overall the environmental and physical factors seemed to have a limited influence on variation in captures among trap locations, with temporal variation in distribution perhaps overwhelming potential influences of local trap conditions.     

Expression of esterases during ontogenesis of the flour beetle Tribolium castaneum (tenebrionidae; coleoptera)

Two electrophoretically fast-migrating, nonspecific esterases were detected in two strains of the flour beetle Tribolium castaneum and designated F (fast) and S (slow) according to their relative migration distances. Both isozymes are associated with the alimentary canal and display ontogenetic changes. Their activity is very low in the egg stage, increases in the larva, and declines dramatically in the pharate pupa and pupa. The overall activity in the pupal stage is low, yet increases gradually throughout this period. In the adult, the activity of the esterases rises sharply. The larval and adult F and S isozymes were found to hydrolyze - and -naphthylacetate and -naphthylpropionate with almost equal capacity. -Naphthyl laurate was cleaved by the F enzyme of both larvae and adults. The F and S were insensitive to inhibitors of arylesterases and cholinesterases and were markedly inhibited by the organophosphate diisopropylphosphorofluoridate (DFP) and could be classified as carboxylesterases. Differential sensitivities of larval and adult esterases to urea and heat treatment as well as to DFP may indicate the expression of different genes during metamorphosis.     

Polyandry and female control: the red flour beetle Tribolium castaneum as a case study

Females of many animal species are polyandrous, and there is evidence that they can control pre- and post-mating events. There has been a growing interest in consequences of polyandry for male and female reproductive success and offspring fitness, and its evolutionary significance. In several taxa, females exhibit mate choice both before and after mating and can influence the paternity of their offspring, enhancing offspring number and quality, but potentially countering male interests. Studying female mating biology and in particular post-copulatory female control mechanisms thus promises to yield insights into sexual selection and the potential of male-female coevolution. Here, we highlight the red flour beetle Tribolium castaneum (Herbst), a storage pest, as a model system to study polyandry, and review studies addressing the effects of polyandry on male sperm competitive ability and female control of post-mating events. These studies show that the outcome of sperm competition in the red flour beetle is influenced by both male and female traits. Furthermore, recent advances suggest that sexual conflict may have shaped reproductive traits in this species.     

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.     

The effects of Farinocystis tribolii on the growth and development of the flour beetle Tribolium castaneum

Farinocystis tribolii multiplied only in the fatbody of Tribolium castaneum larvae. In the advanced stages of infection, the fatbody was destroyed and the albuminoid granules were completely depleted. Larvae in late stages of infection were “C” shaped. Diseased pupae were flat and straight with the head and mouth parts not completely flexed with the pupal body. Infected pupae failed to develop into adults. Heavily infected adults were distended. Larvae of third, fourth, and fifth instars, although succumbing to the infection ultimately, continued to live even after the healthy larvae of the same age had entered pupation, indicating a prolongation of larval period. Molting was either affected or delayed in infected insects. F. tribolii infection induced a juvenile hormonal effect producing larval-pupal and pupal-adult intermediate forms. Ether extract of spores of F. tribolii when applied on pupae of T. castaneum and final instar nymphs of Dysdercus cingulatus produced adultoids.     

Electrophoretic separation of enzymes of individual flour beetles, Tribolium castaneum, on polyacrylamide gel

A method of preparing a single-beetle homogenate for separating enzymes of individual flour beetles, Tribolium castaneum, by electrophoresis on polyacrylamide gel is described. This method can be adapted to the study of enzyme activity in organisms similarf to T. castaneum in size and protein concentration (450 μg dry wt), or for the analysis of very small quantities of tissue and for whole organs from a slightly larger organism.     

Variation between strains of the flour beetle Tribolium castaneum in relative performance on five flours

When populations are exposed to different environments, evolutionary processes can lead either to genetically differentiated strains or to the appearance of increased generalism at the individual level. For evolution to occur, genetic variability in performance in different environments is required. Here, intraspecific genetic variation across environments was estimated in the flour beetle Tribolium castaneum (Herbst) by comparing the responses of two strains of T. castaneum to different flour types. Replicated groups from each strain were allowed to develop on either the standard whole wheat medium or on one of four novel flours (wheat, rice, corn and oat). In several of the novel flours, clear differences in mean development time or population size of one or both strains were seen relative to performance in the standard medium. Moreover, the strains differed significantly in their phenotypic responses to the flours. One strain did particularly poorly on oat flour. Reduced oviposition, reduced larval survivorship and increased larval cannibalism were examined as possible causes of the low productivity on oat flour. These three factors accounted for about 70% of the reduction in population size when this strain oviposited and developed in oat flour. The difference between these two outbred strains in response to these five flours suggests that genetic variation in resource use is present within T. castaneum and may also be present within strains and natural populations in grain storage facilities. Such variation would permit an evolutionary response to selection in multiple environments (flours). This process has agricultural implications when several types of grain are stored in a single location because it could eventually lead to the evolution of highly generalized populations of T. castaneum, an important pest of stored products.     

Relationship between population growth of the red flour beetle Tribolium castaneum and protein and carbohydrate content in flour and starch

The effects of eight diets (atta flour, wheat flour, self-rising flour, rice flour, custard powder, corn flour, tapioca starch, and potato starch) on the development of the red flour beetle, Tribolium castaneum (Herbst), reared at 29-31 degrees C and 66-70% RH were assessed. Five pairs of male and female T. castaneum were reared on the respective diets for 28 d before the experimental setup was dismantled and adult counts were recorded. In another experiment, the insects were allowed to mate and oviposit in each flour or starch type over a period of 7 d before being removed. The counting of pupae and adult emergence began on the day of emergence and was continued on a daily basis until day 140. Proximate analysis was performed for chemical composition of each diet, and the numbers of new adults that developed were found to be positively correlated (r2 = 0.97; P < 0.05) with the protein content and negatively correlated (r2 = 0.93; P < 0.05) with the carbohydrate content. For T. castaneum, the suitable diets were ranked as follows: atta flour > wheat flour > self-rising flour > rice flour > custard powder > corn flour > tapioca starch > potato starch. T. castaneum larval development to the pupal and adult stages developed significantly faster in atta flour (P < 0.05) than in the other diets, and the greatest number of progeny was produced from beetles reared on atta flour. Fewer adults emerged from wheat flour, self-rising flour, and rice flour, and no new emergences were recorded for the remaining diets. Developmental rate was much slower in beetles reared on diets in which a low number in progeny was produced. These data illustrate that different diets can influence the sustainability of these insects and affect their development and growth.     

Harmaline ingestion effect on development,metabolites and midgut of the red flour beetle,Tribolium castaneum

Alkaloids constitute an interesting family of molecules synthesized and accumulated by certain plants and it suggested that they participate in plant defences. In this study, we determined the ingestion effect of the alkaloid, harmaline, on the insect pest Tribolium castaneum Herbst (Coleoptera: Tenebrionidae). When incorporated into the diet at 400, 1000, and 2000 ppm, harmaline provoked larval weight loss, larval mortality and adult emergence inhibition. Spectrophotometric analyses showed that harmaline ingestion caused a significant diminution in the rate of protein, glycogen and lipid. Besides, the activity of the enzyme α-amylase was also significantly reduced in treated larvae. In addition, by using the optical and electron microscopy, we have showed that harmaline ingestion induced a marked cytotoxic effect on the midgut epithelial cells. They exhibited rare microvilli and an extensive disorganization and vacuolisation of the cytoplasm and the nucleus. In conclusion, the results obtained from this study prove and show potent insecticidal effect of Alkaloids such as harmaline on Tribolium castaneum.     

温湿度对赤拟谷盗成虫飞行能力的影响

本文利用昆虫飞行磨研究了温度和湿度变化对赤拟谷盗Tribolium castaneum(Herbst)雌、雄成虫飞行能力的影响;分别测定了23℃、26℃、29℃、32℃和35℃等5个温度,45%、65%和85%等3个湿度条件下,赤拟谷盗雌、雄成虫的飞行距离、时间和速度等反应昆虫飞行能力的参数。单次飞行距离、单次飞行时间、单次飞行速度、24 h累计飞行距离及时间分析结果表明,环境温度和湿度对赤拟谷盗雌、雄成虫的飞行能力有不同程度的影响,环境温湿度过高或过低均不利于赤拟谷盗雌、雄成虫飞行;温度为32℃时,赤拟谷盗雌成虫的单次飞行距离、飞行时间、飞行速度、24 h累计飞行距离和飞行时间分别为0.925 m、9.616 s、0.023 m/s、0.979 km和2.886 h;雄成虫的单次飞行距离、飞行时间、飞行速度、24 h累计飞行距离和飞行时间分别为0.638 m、6.035 s、0.014 m/s、0.674 km和3.175 h。在相对湿度45%、65%和85%时,雌成虫的单次飞行时间分别为6.998 s、9.616 s和6.431 s;雄成虫的单次飞行时间分别为3.163 s、6.035 s和0.208 s。飞行速度与相对湿度的关系与飞行时间相似。从飞行能力参数来看,雌虫的飞行能力优于雄虫。     

Metamorphosis and adult development of the mushroom bodies of the red flour beetle, Tribolium castaneum

The insect mushroom bodies play important roles in a number of higher processing functions such as sensory integration, higher level olfactory processing, and spatial and associative learning and memory. These functions have been established through studies in a handful of tractable model systems, of which only the fruit fly Drosophila melanogaster has been readily amenable to genetic manipulations. The red flour beetle Tribolium castaneum has a sequenced genome and has been subject to the development of molecular tools for the ready manipulation of gene expression; however, little is known about the development and organization of the mushroom bodies of this insect. The present account bridges this gap by demonstrating that the organization of the Tribolium mushroom bodies is strikingly like that of the fruit fly, with the significant exception that the timeline of neurogenesis is shifted so that the last population of Kenyon cells is born entirely after adult eclosion. Tribolium Kenyon cells are generated by two large neuroblasts per hemisphere and segregate into an early-born delta lobe subpopulation followed by clear homologs of the Drosophila gamma, alpha'/beta' and alpha/beta lobe subpopulations, with the larval-born cohorts undergoing dendritic reorganization during metamorphosis. BrdU labeling and immunohistochemical staining also reveal that a proportion of individual Tribolium have variable numbers of mushroom body neuroblasts. If heritable, this variation represents a unique opportunity for further studies of the genetic control of brain region size through the control of neuroblast number and cell cycle dynamics.     

Ecdysteroid regulation of ovarian growth and oocyte maturation in the red flour beetle,Tribolium castaneum

Previous studies from our laboratory showed the involvement of juvenile hormone (JH) and ecdysteroid signaling in the regulation of female reproduction in the red flour beetle, Tribolium castaneum. JH regulates vitellogenin (Vg) synthesis in the fat body but the role of ecdysteroid signaling is not known. Here, we report on ecdysteroid regulation of ovarian growth and oocyte maturation. Microarray analysis of RNA isolated from ovaries showed the up-regulation of several genes coding for proteins involved in ecdysteroid signaling on the 4th day after female adult eclosion. The functional analyses of genes coding for proteins involved in ecdysteroid and JH signaling pathways by RNA interference (RNAi) revealed that ecdysteroids but not JH regulate ovarian growth and primary oocyte maturation. Ultrastructural studies showed the temporal sequences of key events in oogenesis including the development of primary oocytes, the differentiation and development of follicle epithelial cells, and the formation of intercellular spaces to facilitate uptake of Vg protein. RNAi studies showed that ecdysone receptor (EcR) and ultraspiracle (USP) are required for the ovarian growth, primary oocyte maturation and the growth and migration of the follicle cells. These studies suggest important roles for ecdysteroids in the regulation of oocyte maturation in the beetle ovaries.     

The EU mutant of the flour beetle,Tribolium castaneum herbst. Environmental and genetic effects on penetrance

The mutation eu in the flour beetle Tribolium castaneum, causes an easily detectable morphological change in the larva and the pupa. The character shows incomplete penetrance at 30°C and almost full penetrance at 25°C. Penetrance at 30°C appears to be partly under genetic control.