Effects of rearing density, age, sex, and food deprivation on flight initiation of the red flour beetle (Coleoptera: Tenebrionidae)

Effects of rearing density, adult density and sex ratio in the flight chamber, adult age, sex, presence or absence of food, and duration of food deprivation on rate of and time to flight initiation of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), were studied in the laboratory. Rates of flight initiation were slightly lower at lower rearing densities, but they did not differ with age or sex of adults, presence or absence of food in the flight chamber, or duration of food deprivation. Focal adults were less likely to fly when individuals of the opposite sex were present in flight chambers. Presence of the same sex or mixed sexes and numbers of individuals in flight chambers did not affect tendency to fly of focal individuals. Mean time to flight of older beetles (7-20 d old) was less than that of younger beetles (1-4 d old). No young beetles flew during the first 24 h of flight tests. Mean time to flight did not differ with rearing density, sex, presence or absence of food, or duration of food deprivation. Our results indicate that this species is highly dispersive during the adult stage and that flight does not seem to be associated with just prereproductive or postreproductive dispersal phases, high rearing density, or short to medium periods of food deprivation. Therefore, T. castaneum level of flight activity does not seem to be associated with factors that have been shown in the literature to increase dispersal by walking for this species and to increase flight initiation in other stored-product species.     

Thermal death kinetics of red flour beetle (Coleoptera: Tenebrionidae)

While developing radio frequency heat treatments for dried fruits and nuts, we used a heating block system developed by Washington State University to identify the most heat-tolerant life stage of red flour beetle, Tribolium castaneum (Herbst), and to determine its thermal death kinetics. Using a heating rate of 15 degrees C/min to approximate the rapid heating of radio frequency treatments, the relative heat tolerance of red flour beetle stages was found to be older larvae > pupae and adults > eggs and younger larvae. Lethal exposure times for temperatures of 48, 50, and 52 degrees C for the most heat-tolerant larval stage were estimated using a 0.5th order kinetic model. Exposures needed for 95% mortality at 48 degrees C were too long to be practical (67 min), but increasing treatment temperatures to 50 and 52 degrees C resulted in more useful exposure times of 8 and 1.3 min, respectively. Red flour beetle was more sensitive to changes in treatment temperature than previously studied moth species, resulting in red flour beetle being the most heat-tolerant species at 48 degrees C, but navel orangeworm, Amyelois transitella (Walker), being most heat tolerant at 50 and 52 degrees C. Consequently, efficacious treatments for navel orangeworm at 50-52 degrees C also would control red flour beetle.     

Electroantennographic responses of red flour beetle Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) to volatile organic compounds

The red flour beetle Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is the most encountered and destructive stored product insect pest of cereal grains and seeds. Although this beetle has been used as a model organism for many decades, there is no systematic knowledge about antennal detection of host and non‐host volatiles. Electroantennogram responses to 94 selected volatile organic compounds including alkanes, alkenes, alcohols, aldehydes, ketones, carboxylic acids, esters, terpenoids and aromatic compounds were recorded from both sexes of T. castaneum. Overall, female and male T. castaneum exhibited similar electroantennography (EAG) responses. Compounds eliciting the strongest EAG responses within compound groups of chemical similarity were undecane, 1‐hexen‐3‐ol, octanal, 2‐heptanone, hexanoic acid and ethyl hexanoate. Comparison of vapour pressure and EAG amplitudes within homologous series of compounds revealed responses to undecane, octadecane, octanal, nonanal, 2‐heptanone, hexanoic acid and octanoic acid as outstanding. Given that systematic EAG screenings have not been conducted before, these are the best candidates for evaluation in future behavioural studies to unravel their potential for application in integrated pest management strategies of T. castaneum.     

Compensatory proteolytic responses to dietary proteinase inhibitors in the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae)

Increasing levels of inhibitors that target cysteine and/or serine proteinases were fed to Tribolium castaneum larvae, and the properties of digestive proteinases were compared in vitro. Cysteine proteinases were the major digestive proteinase class in control larvae, and serine proteinase activity was minor. Dietary serine proteinase inhibitors had minimal effects on either the developmental time or proteolytic activity of T. castaneum larvae. However, when larvae ingested cysteine proteinase inhibitors, there was a dramatic shift from primarily cysteine proteinases to serine proteinases in the proteinase profile of the midgut. Moreover, a combination of cysteine and serine proteinase inhibitors in the diet prevented this shift from cysteine proteinase-based digestion to serine proteinase-based digestion, and there was a corresponding substantial retardation in growth. These data suggest that the synergistic inhibitory effect of a combination of cysteine and serine proteinase inhibitors in the diet of T. castaneum larvae on midgut proteolytic activity and beetle developmental time is achieved through the prevention of the adaptive proteolytic response to overcome the activity of either type of inhibitor.     

Insecticidal potential of natural zeolite and diatomaceous earth formulations against rice weevil (Coleoptera: Curculionidae) and red flour beetle (Coleoptera: Tenebrionidae)

Insecticidal potential of natural zeolites and diatomaceous earths originating from Serbia against Sitophilus oryzae (L.) and Tribolium castaneum (Herbst) was evaluated. Two natural zeolite formulations (NZ and NZ Modified) were applied to wheat at rates of 0.50, 0.75, and 1.0 g/kg, while two diatomaceous earth (DE) formulations (DE S-1 and DE S-2) were applied at rates of 0.25, 0.50, 0.75, and 1.0 g/kg. A bioassay was conducted under laboratory conditions: temperature of 24 +/- 1 degrees C, relative humidity in the range 50-55%, in tests with natural zeolites, and 60-65%, in tests with DEs, and in all combinations for progeny production. Mortality was assessed after 7, 14, and 21 d of insect contact with treated wheat, and the total mortality after an additional 7-d recovery on untreated broken wheat. Progeny production was also assessed after 8 wk for S. oryzae and 12 wk for T. castaneum. The highest mortality for S. oryzae and T. castaneum was found after the longest exposure period and 7 d of recovery, on wheat treated with NZ at the highest rate and DEs at rates of 0.50 -1.0 g/kg. Progeny reduction higher than 90% was achieved after 14 and 21 d of contact of both beetle pests with wheat treated with DE S-1 at 0.50-1.0 g/kg and DE S-2 at 0.75-1.0 g/kg, while the same level of reduction was achieved only for T. castaneum after its contact with the highest rate of NZ formulation. NZ Modified, applied even at the highest rate, revealed much lower insecticidal potential.     

Effects of proteinase inhibitors on digestive proteinases and growth of the red flour beetle,Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)

The physiology of the gut lumen of the red flour beetle, T. castaneum, was studied to determine the conditions for optimal protein hydrolysis. Although the pH of gut lumen extracts from T. castaneum was 6.5, maximum hydrolysis of casein by gut proteinases occurred at pH 4.2. The synthetic substrate N-alpha-benzoyl-DL-arginine-rho-nitroanilide was hydrolyzed by T. castaneum gut proteinases in both acidic and alkaline buffers, whereas hydrolysis of N-succinyl-ala-ala-pro-phe rho-nitroanilide occurred in alkaline buffer. Inhibitors of T. castaneum digestive proteinases were examined to identify potential biopesticides for incorporation in transgenic seed. Cysteine proteinase inhibitors from potato, Job's tears, and sea anemone (equistatin) were effective inhibitors of in vitro casein hydrolysis by T. castaneum proteinases. Other inhibitors of T. castaneum proteinases included leupeptin, L-trans-epoxysuccinylleucylamido [4-guanidino] butane (E-64), tosyl-L-lysine chloromethyl ketone, and antipain. Casein hydrolysis was inhibited weakly by chymostatin, N-tosyl-L-phenylalanine chloromethyl ketone, and soybean trypsin inhibitor (Kunitz). The soybean trypsin inhibitor had no significant effect on growth when it was bioassayed alone, but it was effective when used in combination with potato cysteine proteinase inhibitor. In other bioassays with single inhibitors, larval growth was suppressed by the cysteine proteinase inhibitors from potato, Job's tears, or sea anemone. Levels of inhibition were similar to that observed with E-64, although the moles of proteinaceous inhibitor tested were approximately 1000-fold less. These proteinaceous inhibitors are promising candidates for transgenic seed technology to reduce seed damage by T. castaneum.     

Digestive proteinases of the larger black flour beetle, Cynaeus angustus (Coleoptera: Tenebrionidae)

Digestion in the larger black flour beetle, Cynaeus angustus (LeConte), was studied to identify new control methods for this pest of stored grains and grain products. The physiological pH of the larval gut, as measured with extracts in water, was approximately 6.1, and the pH for optimal hydrolysis of casein by gut extracts was 6.2 when buffers were reducing. However, under non-reducing conditions, hydrolysis of casein and synthetic serine proteinase substrates was optimal in alkaline buffer. Three major proteinase activities were observed in zymograms using casein or gelatin. Caseinolytic activity of C. angustus gut extracts was inhibited by inhibitors that target aspartic and serine proteinase classes, with minor inhibition by a cysteine proteinase inhibitor. In particular, soybean trypsin and trypsin/chymotrypsin inhibitors were most effective in reducing the in vitro caseinolytic activity of gut extracts. Based on these data, further studies are suggested on the effects of dietary soybean inhibitors of serine proteinases, singly and in combination with aspartic and cysteine proteinase inhibitors, on C. angustus larvae. Results from these studies can be used to develop new control strategies to prevent damage to grains and stored products by C. angustus and similar coleopteran pests.     

Impact of food source on survival of red flour beetles and confused flour beetles (Coleoptera: Tenebrionidae) exposed to diatomaceous earth

A series of experiments was conducted to determine the effect of a flour food source on survival of red flour beetle, Tribolium castaneum (Herbst), and confused flour beetle, Tribolium confusum (DuVal), exposed to the labeled rate (0.5 mg/cm2) of Protect-It, a marine formulation of diatomaceous earth. Beetles were exposed at 27 degrees C, and 40, 57, and 75% RH in 62-cm2 petri dishes. When beetles were exposed for 1 or 2 d in dishes with the labeled rate (0.5 mg/cm2, or 31 mg per dish) of diatomaceous earth or in dishes containing flour at varying levels from 0 to 200 mg mixed with the labeled rate of diatomaceous earth, survival of both species increased as the amount of flour increased, and quickly plateaued at levels approaching 100%. In a second set of experiments, beetles were transferred to dishes containing flour at varying levels from 0 to 200 mg after they were exposed for 1 or 2 d in dishes with the labeled rate of diatomaceous earth alone. There were no significant differences in beetle survival among the levels of flour, however, survival in dishes with flour was usually greater than survival in dishes with diatomaceous earth alone. In a third test, beetles were exposed for 1, 2, and 3 d in dishes with either the labeled rate of diatomaceous earth alone (clean dishes), dishes with diatomaceous earth and empty straws, or dishes with diatomaceous earth and approximately 300 mg of flour packed in the straws. Survival was not significantly different between clean dishes or dishes with straws, but survival in dishes containing the straws with flour was usually 100%, regardless of exposure interval. In all experiments, confused flour beetles were less susceptible to diatomaceous earth than red flour beetles. In addition, survival was negatively related to exposure interval and positively related to relative humidity.     

Phosphine resistance in the rust red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae): inheritance, gene interactions and fitness costs

The recent emergence of heritable high level resistance to phosphine in stored grain pests is a serious concern among major grain growing countries around the world. Here we describe the genetics of phosphine resistance in the rust red flour beetle Tribolium castaneum (Herbst), a pest of stored grain as well as a genetic model organism. We investigated three field collected strains of T. castaneum viz., susceptible (QTC4), weakly resistant (QTC1012) and strongly resistant (QTC931) to phosphine. The dose-mortality responses of their test- and inter-cross progeny revealed that most resistance was conferred by a single major resistance gene in the weakly (3.2×) resistant strain. This gene was also found in the strongly resistant (431×) strain, together with a second major resistance gene and additional minor factors. The second major gene by itself confers only 12-20× resistance, suggesting that a strong synergistic epistatic interaction between the genes is responsible for the high level of resistance (431×) observed in the strongly resistant strain. Phosphine resistance is not sex linked and is inherited as an incompletely recessive, autosomal trait. The analysis of the phenotypic fitness response of a population derived from a single pair inter-strain cross between the susceptible and strongly resistant strains indicated the changes in the level of response in the strong resistance phenotype; however this effect was not consistent and apparently masked by the genetic background of the weakly resistant strain. The results from this work will inform phosphine resistance management strategies and provide a basis for the identification of the resistance genes.     

Effect of Tribolium castaneum (Coleoptera: Tenebrionidae) nutritional environment, sex, and mating status on response to commercial pheromone traps

Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), the red flour beetle, is an important cosmopolitan pest of stored grains. Commercial traps baited with the synthetic aggregation pheromone 4,8-dimethyldecanal (DMD) are used to monitor T. castaneum population densities in storage facilities. However, trap catches may depend on several intrinsic and extrinsic factors. In this study, we explored the effects of beetle nutritional environment, sex, and mating status on the response of T. castaneum to commercial Storgard Dome traps. Beetles raised on a low-nutrition diet were 1.7 times more likely to enter DMD-baited traps compared with beetles that were raised on a high-nutrition diet. Although no sex difference in trap response was found, unmated beetles of both sexes were more responsive to DMD than were mated beetles, and this effect was especially pronounced for beetles reared on a low-nutrition diet. These results suggest that estimating T. castaneum population densities based on trap catches might be improved by incorporating information about the nutritional quality of infested stored products.     

Polymorphism observed in mitochondrial genes of red flour beetle,Tribolium castaneum (Coleoptera: Tenebrionidae) of different origin in laboratory cultures

Tribolium castaneum, a pest of stored grain, has a uniform morphology, preventing the visual identification of strains from different areas. Polymorphisms in the nucleotide sequences of the mitochondrial genes of this species were examined, and combined into seven haplotypes among the test insect specimens originating from Japan, Thailand, and Canada. These results suggested the potential for geographical differentiation.     

Novel molecular identification methods for the larger black flour beetle,Cynaeus angustus (Coleoptera: Tenebrionidae)

Two new methods were developed for identifying Cynaeus angustus (LeConte) (Coleoptera: Tenebrionidae) by DNA amplification using simplex and real-time PCR targeting the cytochrome c oxidase subunit I (COI) sequence reported previously. The specificities of the PCR primers and probe were also confirmed by the two PCR methods using the 22 main stored-product insect species, including DNA samples from nine tenebrionid beetle species. The results showed that the newly developed simplex and real-time PCR-based methods have sufficient specificity for analysis. The limits of detection for C. angustus total DNA by the simplex and multiplex PCR were 320 fg and 20 pg, respectively.

     

Influence of age at mating on the reproductive performance of Parthenium beetle,Zygogramma bicolorata (Coleoptera: Chrysomelidae)

Abstract Age-specific mating incidence, sexual maturation and effect of age at mating on reproductive performance of the Parthenium beetle, Zygogramma bicolorata Pallister, was studied. Based on 50% mating incidence the calculated age of sexual maturation of males and females was 10.5 and 11.1 days, respectively, which was not statistically significant. However, on the basis of age at first mating, that is, sexual maturity, females matured 2 days earlier than males. Fecundity, pre-oviposition, oviposition and post-oviposition period and female longevity appear to be influenced by female age at mating with reproductive performance peaking at 30 days. On the other hand, egg viability was influenced by male age and was highest when males mated at the age of 40 days. To summarise, egg production and timing of egg deposition was female age-dependent, whereas egg fertility was male age-dependent. It was also observed that females mated at a later age and laid a higher number of eggs immediately after mating than did earlier mated females. This was ostensibly in a bid to increase fitness by maximizing reproductive output in the reduced life span available. This is the first investigation on the effect of age of females at mating on reproduction in this beetle.     

Susceptibility of last instar red flour beetles and confused flour beetles (Coleoptera: Tenebrionidae) to hydroprene.

Last instar larvae of the red flour beetle, Tribolium castaneum (Herbst), and the confused flour beetle, Tribolium confusum Jacquelin du Val, were either exposed for 8-144 h on concrete treated with 1.9 x 10(-3) mg(AI)/per cm2 hydroprene, or continually exposed on concrete treated with 9.8 x 10(-4) to 1.9 x 10(-3) mg[AI]/per cm2 hydroprene. In both tests, larvae were exposed and held at 27 or 32 degrees C and 40, 57, or 75% RH. When larvae were exposed with no food to hydroprene for different time intervals, then transferred to untreated concrete containing flour, consistent effects were produced only at 144 h. At this exposure interval, the percentage of beetles arrested in the larval stage after 3-4 wk was generally greater at 75% RH compared with 40 and 57% RH, but there were no differences between species or temperature. The percentages of dead adult red flour beetles and live adults with morphological deformities were also greatest at 75% RH, and defects were more prevalent in red flour beetles than in confused flour beetles. When larvae were continually exposed to different concentrations of hydroprene on concrete that contained flour, the percentage of arrested larvae, dead adults, and live adults of both species generally increased with concentration. There were more deleterious effects at 75% RH compared with either 40 or 57% RH, and effects were more pronounced in the red flour beetle compared with the confused flour beetle. In both experiments, temperature effects were variable and inconclusive. Results indicate that continual exposure of last instar red flour beetle and confused flour beetle to hydroprene can limit population development, but exposure intervals of >6 d may be required for maximum effectiveness.     

Fitness consequences of malathion-specific resistance in red flour beetle (Coleoptera: Tenebrionidae) and selection for resistance in the absence of malathion

Malathion resistance in the red flour beetle, Tribolium castaneum (Herbst), is a worldwide problem and is very stable once it becomes widespread in natural populations. In the absence of insecticide the proportion of resistant phenotypes may rapidly decline but the development of resistance does not always involve reduced fitness. Malathion-specific resistance in T. castaneum seems not to involve any loss of fitness in laboratory or field conditions. Susceptible beetles were in competition with resistant beetles at different initial frequencies and modifications of susceptible gene frequency were estimated in these laboratory populations over 10 generations. A significant decrease in susceptible gene frequency was observed in Tribolium populations over time. The selection coefficient of the susceptible allele was estimated and the fitness of susceptible alleles in all tests was observed to range from 0.89 to 0.93 compared with the fitness of resistant genotypes, which was assumed to be 1. Data provided evidence that the resistant strains exhibited fitness advantages in the absence of malathion. We also compared the biotic potential (fecundity and developmental time) of the susceptible strain, the homozygous malathion-specific resistant strain, and their hybrids. Malathion-specific resistant strains showed an 8 -23% increase in biotic potential relative to the susceptible strain. These findings are consistent with those of malathion-specific resistance in T. castaneum; the fitness of the insects seems independent of the genetic background and the fitness of the resistant insects is not affected by this resistance mechanism.     

The early stages of ommatidial development in the flour beetle Tribolium castaneum (Coleoptera; Tenebrionidae)

 Using electron microscopy, the first stages of ommatidial development in the flour beetle Tribolium castaneum were analysed in relation to the cellular architecture of the adult compound eye and were compared to the corresponding patterning process in the fruit fly Drosophila melanogaster. The ommatidia of the slightly horse-shoe shaped beetle compound eye contain six peripheral and two central retinula cells. The rhabdomere of the posteriorly located central photoreceptor cell is restricted to the distal half of the rhabdom whilst that of the anterior one is restricted to its proximal half. The development of the compound eye takes place in an external eye imaginal disc. Most stages of ommatidial development, as known from Drosophila, i.e. arc-like cell groups, five-cell clusters, immature eight-cell clusters and symmetrical eight-cell clusters, are very precisely conserved between the two species. Two major differences exist: 1. In Tribolium, the cone cell precursor cells synchronously join to the immature eight-cell cluster. As a consequence, the symmetrical eight-cell cluster immediately transforms into a four-cone-cell cluster. 2. The maturing ommatidia do not undergo rotation in Tribolium. Overall, no morphological indiation for an equator in the adult Tribolium compound eye could be found. Considering the strong evolutionary conservation of early ommatidial development, homology of photoreceptor cells of distantly related insects is proposed to be inferred from their ontogenetic origin. Received: 6 November 1995 / Accepted: 9 April 1996     

Dormancy-inducing factors of rubber litter beetle, Luprops tristis (Coleoptera: Tenebrionidae)

Abstract  The effect of photoperiod, temperature, humidity, rainfall and moisture were examined on the annual migration of Luprops tristis (Fabricius, 1801) (Coleoptera: Tenebrionidae) from rubber plantations to shelters, and the induction of 9 months dormancy thereafter. Results confirm the role of rainfall-mediated wetness from summer showers as the cue that induces dormancy in Luprops tristis and provide further proof to the function and significance of moisture in diapause induction in tropical insects. Other factors did not have any significant independent effect on migration and dormancy induction. Reserve food accumulation by teneral adults well ahead of the onset of adverse conditions and the non-return of beetles to the field in short-term favorable conditions strongly indicate that Luprops tristis has evolved the ability to "predict" and avoid adverse conditions and the inheritance of dormancy-regulating factors from the parent generation.