The spatiotemporal dynamics of Tribolium castaneum (Herbst): adult flight and gene flow

Tribolium castaneum (Herbst) has been used as a model organism to develop and test important ecological and evolutionary concepts and is also a major pest of grain and grain products globally. This beetle species is assumed to be a good colonizer of grain storages through anthropogenic movement of grain, and active dispersal by flight is considered unlikely. Studies using T. castaneum have therefore used confined or walking insects. We combine an ecological study of dispersal with an analysis of gene flow using microsatellites to investigate the spatiotemporal dynamics and adult flight of T. castaneum in an ecological landscape in eastern Australia. Flying beetles were caught in traps at grain storages and in fields at least 1 km from the nearest stored grain at regular intervals for an entire year. Significantly more beetles were trapped at storages than in fields, and almost no beetles were caught in native vegetation reserves many kilometres from the nearest stored grain. Genetic differentiation between beetles caught at storages and in fields was low, indicating that although T. castaneum is predominantly aggregated around grain storages, active dispersal takes place to the extent that significant gene flow occurs between them, mitigating founder effects and genetic drift. By combining ecological and molecular techniques, we reveal much higher levels of active dispersal through adult flight in T. castaneum than previously thought. We conclude that the implications of adult flight to previous and future studies on this model organism warrant consideration.     

The development of PCR-based markers for molecular sex identification in the model insect species Tribolium castaneum

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is a major pest of stored grain and cereal crops. It is also an important model species in genetic, ecological, and evolutionary research. The majority of its genome was recently sequenced and published. However, the genomic sequence of the small Y-chromosome is still undetermined, which hinders the development of molecular sex identification methods. Traditional methods for sexing adult forms of Tribolium beetles are troublesome. Therefore, a method for molecular sex identification in the red flour beetle was developed. One sex-linked randomly amplified polymorphic DNA marker was converted into a sequence-characterized amplified region (SCAR). The SCAR was aligned with the T. castaneum reference whole-genome sequence and fully matched a fragment of a single contig of unknown genomic location. The novelty of the method is that the fragment consists of shorter DNA fragments that are also present at other locations around the genome, but the particular order of these fragments within the sequenced region appeared to be Y-specific and this property was utilized for marker development. A set of three primers for multiplex PCR reaction was designed resulting in amplification of different length Y-specific and not-Y-specific (control) DNA fragments in a single PCR, which allows to distinguish males from females. The primers successfully sexed pre-sexed pupae and adult beetles from six laboratory strains, showing that the order of the repeated fragments is conserved in the species and is not strain-specific.     

Efficacy of Beauveria bassiana for control of Tribolium castaneum with reduced oxygen and increased carbon dioxide

This study investigated the effect of atmosphere modification, a widely adopted means of insect control in stored products, on the efficacy of Beauveria bassiana for one of the most difficult to control pests, Tribolium castaneum. Oxygen reduction to 5% (±1%) as opposed to CO₂ elevation to 40% (±2%) for the first 72 h of fungus exposure resulted in significantly greater larval mortality than fungus exposure under ambient atmospheres. Both treatments reduced pupation of older larvae suggesting that slowed development may be a beneficial factor for fungal efficacy. CO₂ elevation but not O₂ reduction significantly affected the mortality of adult beetles that were exposed to the fungus. Carbon dioxide elevation significantly reduced B. bassiana’s germination and growth rates, but oxygen reduction did not.     

Permethrin- and malathion-induced macromolecular abnormalities in adult Tribolium castaneum (herbst)

In Tribolium castaneum adults, sublethal doses of 1 and 2 ppm permethrin and 300 ppm malathion led to significant changes in amylase, trehalase, acid phosphatase, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and isocitrate dehydrogenase activities. Malathion at 150 ppm did not affect phosphatases and lactate dehydrogenase. Both malathion and permethrin significantly increased cholinesterase activity. Mixing of the two insecticides resulted in antagonistic action with reference to various enzymatic activities. Glucose and glycogen contents were at first mobilized for energy supply under insecticidal stress conditions followed by lipid and cholesterol. Soluble protein, total protein, free amino acids, and urea contents remained unaltered under all experimental conditions.     

Responses of Tribolium castaneum to olfactory cues from cotton seeds,the fungi associated with cotton seeds,and cereals

We tested, in an olfactometer, whether or not Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) responds preferentially to the volatiles that emanate from the fungi associated with cotton [Gossypium hirsutum L. (Malvaceae)] seed over those that emanate from cereals, because cereals are usually portrayed as the primary resources of these beetles. Pairwise comparisons were conducted between cotton seed, wheat (Triticum aestivum L.), and sorghum [Sorghum bicolor (L.) Moench] (both Poaceae); volatiles were tested from intact seeds and from both water and ethanol extracts. The results demonstrate that T. castaneum is attracted more strongly to cotton seeds with its lint contaminated with fungi, than to the conventional resources of this species (i.e., wheat and sorghum). Further tests prove that it is the fungus on the lint that produces the active volatiles, because the beetles did not respond to sterilized cotton lint (i.e., without the fungi typically associated with it when cotton seed is stored). Tests with five fungal cultures (each representing an unidentified species that was isolated from the field‐collected cotton lint) were variable across the cultures, with only one of them being significantly attractive to the beetles. The others were not attractive and one may even have repulsed the beetles. The results are consistent with the beetles having a strong ecological association with fungi and suggest it would be worth investigating the ecology of T. castaneum from this perspective.     

Effects of caffeine on mating behavior and sperm precedence in Tribolium castaneum

Biogenic amines such as dopamine are physiologically neuroactive substances that affect behavioral and physiological traits in invertebrates, and it has long been known that these substances affect mating behavior in insects. Caffeine is a dopamine activator and thus enhances dopamine receptor activity. However, the effects of caffeine intake on insect mating behavior have been largely unexplored. Therefore, we examined the effect of caffeine on mating behavior in the red flour beetle Tribolium castaneum. Caffeine, which activates dopamine, affected the mating behavior of T. castaneum males. Males who orally ingested caffeine courted faster than males who did not, resulting in faster mounting of females and less time to a male's external aedeagus protrusion. However, the present results showed no difference in sperm precedence measured as a P2 value between males fed caffeine and males not fed caffeine. We discuss the effects of caffeine on insect mating and the possibility that caffeine consumption may cause males to mate with more females in the laboratory.     

Inheritance pattern and expression of resistance to phosphine in larval stage of Tribolium castaneum (Coleoptera: Tenebrionidae)

Resistance developed by Tribolium castaneum (Herbst), a major pest of stored grain and grain products, to phosphine is a serious concern. Treatment of phosphine is given to all the stages (egg, larva, pupa and adult) of insects simultaneously in the infected commodity. Effective resistance management tactics need a proper understanding of expression of resistance in relation to dosage in immature stages including larvae. So, this study was undertaken to understand the genetic basis of phosphine resistance in larval stage of T. castaneum by crossing the resistant and susceptible parents to obtain the F₁, F₂ and the back-cross progenies. The resistant parent had a resistance ratio of 37.75 when compared to the susceptible parent. The susceptibility levels of the reciprocal F₁ hybrids did not show any significant difference indicating autosomal mode of inheritance of resistance. Level of dominance for the reciprocal F₁ hybrids was estimated (-0.70 and ?0.68) which showed that the resistance is expressed as incompletely recessive in larval stage of T. castaneum. χ² analysis showed that the observed and expected mortalities differed significantly (P < 0.05) in most of the concentrations indicating that more than one gene is responsible for phosphine resistance. Two plateaus at ～ 40 % and ～ 90 % mortality levels for the back-cross progeny to the resistant parent further confirmed the involvement of two or more genes in imparting resistance to phosphine. Our results will aid in enhancing the knowledge on the development of resistance to phosphine in the field and thus facilitate in designing suitable resistance management tactics.     

Effects of two pheromone trap densities against banana weevil, Cosmopolites sordidus, populations and their impact on plant damage in Uganda

Abstract:  An on-farm study to evaluate the effect of pheromone trap density on the population of the banana weevil, Cosmopolites sordidus (Germar) (Col., Curculionidae) was conducted in Masaka district, Uganda. The pheromone used was Cosmolure+, a commercially available weevil aggregation pheromone. Forty-two farms were assigned to one of three treatments: 0, 4 and 8 pheromone traps/ha. Pheromone lures were changed monthly at which time the traps were moved to a different location within the stand. Adult weevil population densities were estimated by using mark and recapture methodology at 0, 6, 12, 18 and 21 months, while damage to the banana corm was assessed at 0, 3, 6, 12, 18 and 21 months since the start of the experiment. Pheromone trap captures were generally low: about 10 weevils per trap per month. There were no significant differences in mean catches of C. sordidus per trap per month except for February 2002 when doubling the pheromone trap density decreased weevil catches. Although not significant, decreased efficiency was also the trend in higher trap densities over all the data sets. Doubling the number of traps increased the number of weevils caught per hectare per month from 0.4 to 0.6%. There was no significant difference in plant damage between the pheromone treatments in low- compared with high-trap densities. There were generally no significant differences in weevil populations and plant damage between pheromone-treated and control farms. Possible reasons for the low-trap efficacy in this study are discussed.     

Pre‐ and postcopulatory sexual selection act in concert to determine male reproductive success in Tribolium castaneum

To capture how sexual selection shapes male reproductive success across different stages of reproduction in Tribolium castaneum (Coleoptera, Tenebrionidae), we combined sequential sperm defence (P1) and sperm offence (P2) trials with additional trials where both males were added simultaneously to the female. We found a positive correlation between the relative paternity share in simultaneous male–male competition trials and the P2 trial. This suggests that males preferred by females as sires achieve superior fertilization success during sperm competition in the second male position. In simultaneous male–male competition trials, where pre‐, peri‐ and postcopulatory sexual selection were all allowed to act, the relative paternity share of preferred males was more than 20% higher than in P2 sperm competition trials where precopulatory female choice was disabled. Additional behavioural observations revealed that mating with more attractive males resulted significantly more frequently in offspring production than mating with less attractive males. Thus, by comparing male fertilization success in trials where precopulatory choice was turned off with more inclusive estimates of fertilization success where pre‐ and pericopulatory choice could occur, we show that female mate choice may effectively inhibit sperm competition. Female mate choice and sperm competition (P2) are positively correlated, which is consistent with directional sexual selection in this species. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 67–75.     

Trade‐off between attraction to aggregation pheromones and repellent effects of spike lavender essential oil and its main constituent linalool in the flour beetle Tribolium confusum

Essential oils of aromatic plants and their individual volatile components have been tested in pest management strategies for their toxic and often repellent effects on target insects. When evaluating their possible effects on crucial behaviours of the pest insects, the olfactory environment including intraspecific communication cues has to be considered. We used the flour beetle Tribolium confusum du Val (Coleoptera: Tenebrionidae), a common stored‐product pest, to investigate the influence of oil of spike lavender, Lavandula spica Medik. (Labiaceae), and its main component, linalool, at various doses on olfactory‐guided behaviour. Using four‐way olfactometers, a dose‐dependent repellent effect of L. spica oil and linalool alone was revealed. On the other hand, we confirmed that T. confusum is attracted by conspecifics, by means of an aggregation pheromone and by 10 ng of one of its components, 1‐tetradecene. Twenty‐four hour pre‐exposure to 10 μl of L. spica oil abolished subsequent attraction to 1‐tetradecene and reduced attraction to five conspecifics. Simultaneous exposure to L. spica oil or linalool and five conspecifics reduced the repellent effect of the volatiles in a dose‐dependent manner, whereas simultaneous exposure to 1‐tetradecene at 10 ng abolished the repellent effect of L. spica oil only at a dose of 0.01 mg. These results indicate a dose‐dependent trade‐off between attractive and plant‐derived repellent volatiles, which may influence the effectiveness of such volatiles in their potential use in alternative pest management strategies.     

Real‐time cell analysis and heat shock protein gene expression in the TcA Tribolium castaneum cell line in response to environmental stress conditions

The rust red flour beetle, Tribolium castaneum (Herbst, 1797) (Coleoptera: Tenebrionidae), is a pest of stored grain and one of the most studied insect model species. Some of the previous studies involved heat response studies in terms of survival and heat shock protein expression, which are regulated to protect other proteins against environmental stress conditions. In the present study, we characterize the impedance profile with the xCELLigence Real‐Time Cell Analyzer and study the effect of increased temperature in cell growth and viability in the cell line BCIRL‐TcA‐CLG1 (TcA) of T. castaneum. This novel system measures cells behavior in real time and is applied for the first time to insect cells. Additionally, cells are exposed to heat shock, increased salinity, acidic pH and UV‐A light with the aim of measuring the expression levels of Hsp27, Hsp68a, and Hsp83 genes. Results show a high thermotolerance of TcA in terms of cell growth and viability. This result is likely related to gene expression results in which a significant up‐regulation of all studied Hsp genes is observed after 1 h of exposure to 40 °C and UV light. All 3 genes show similar expression patterns, but Hsp27 seems to be the most affected. The results of this study validate the RTCA method and reveal the utility of insect cell lines, real‐time analysis and gene expression studies to better understand the physiological response of insect cells, with potential applications in different fields of biology such as conservation biology and pest management.     

An attract-and-kill system to control Carpophilus spp. in Australian stone fruit orchards

During two growing seasons, the use of an attract-and-kill system for control of Carpophilus spp. (Coleoptera: Nitidulidae) and the effective range or drawing power of the attract-and-kill stations were examined in stone fruit orchards in the Goulburn Valley, northern Victoria, Australia. Three attract-and-kill stations, baited with synthetic aggregation pheromone plus co-attractant, were placed about 50 m apart in the upwind corner of each treated block 5–6 weeks before the fruit began to ripen. Large numbers of Carpophilus spp. were caught in the attract-and-kill stations immediately after placement. By the time fruit had ripened, the number of Carpophilus spp. caught had decreased greatly. Fruit damage caused by Carpophilus spp. in treated blocks, especially in 2000–2001 season, was almost zero (0.1% and 0.6%) in trees and on the ground, respectively, whereas the damage levels in control blocks were 5.2% and 19.9% in trees and on the ground, respectively. This study indicates that excellent protection of ripening stone fruit may be achieved by using attract-and kill-stations.     

Tree species identity and forest composition affect the number of oak processionary moth captured in pheromone traps and the intensity of larval defoliation

1. Pure forests are often seen as being more prone to damage by specialist pest insects than mixed forests, and particularly mixed forests associating host and nonhost species. We addressed the effect of tree diversity on oak colonization and defoliation by a major specialist pest, the oak processionary moth (OPM)
2. We quantified the number of male OPM moths captured and larval defoliation in pure stands of two oak host species (Quercus robur and Quercus petraea) and in mixed stands associating the two oak species or each oak species with another nonhost broadleaved species. We conducted two complementary studies to test the effect of host species and stand composition: (i) we used pheromone trapping to compare the number of males OPM captured throughout the distribution of oak hosts in France and (ii) we noted the presence of OPM nests and estimated defoliation in mature forests of north‐eastern France.
3. Oak species and stand composition significantly influenced the number of male OPM captured and defoliation by OPM larvae. Quercus petraea was consistently more attractive to and more defoliated by OPM than Q. robur. Both oak trees were attacked more in pure stands than in mixed stands, in particular mixed stands associating oaks with another (nonhost) broadleaved species.
4. The results of the present study support the view that mixed forests are more resistant to specialist pest insects than pure stands, and also indicate that this trend depends on forest composition. Our study provides new insights into OPM ecology and has potential implications for forest management, including the management of urban forests where OPM causes serious human health issues.

     

Mark–recapture of Cerambyx welensii in dehesa woodlands: dispersal behaviour,population density,and mass trapping efficiency with low trap densities

The longhorn beetle, Cerambyx welensii Küster (Coleoptera: Cerambycidae), is an emerging pest involved in oak decline episodes in dehesa open woodlands. Larvae are xylophagous and cause considerable physiological, mechanical, and structural damages to trees. Chemical and biological control are currently unsatisfactory. Recent research has shown that mass trapping with a high density of baited traps (40 traps ha^?1) could be useful to manage C. welensii populations, although such a trap density was too high to be cost‐effective. In this 2‐year study (2010–2011) we investigated with mark–recapture methods in a large plot (1) the flight dispersal behaviour, (2) the adult population density, and (3) the efficiency of mass trapping at two low trap densities (one or four traps ha^?1). Results indicated that many adults were sedentary (60%) but flying adults displayed a strong propensity to move, both sexes dispersing on average more than 200 m and one male and one female flying at least 540 and 349 m, respectively. Recapture rates were high (0.26–0.35) and population density was estimated to be 6–22 adults ha^?1 with maximum likelihood models. Trapping efficiency ranged 48–61% with no significant effect of trap density or year. We conclude that results were not satisfactory enough to recommend mass trapping with low trap densities as control method for C. welensii and that more research is still required on the technical, ecological, and behavioural factors affecting control efficiency.     

Comparison of pheromone trap design and lures for Spodoptera frugiperda in Togo and genetic characterization of moths caught

Fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), is a pest of grain and vegetable crops endemic to the Western Hemisphere that has recently become widespread in sub‐Saharan Africa and has appeared in India. An important tool for monitoring S. frugiperda in the USA is pheromone trapping, which would be of value for use with African populations. Field experiments were conducted in Togo (West Africa) to compare capture of male fall armyworm using three commercially available pheromone lures and three trap designs. The objectives were to identify optimum trap × lure combinations with respect to sensitivity, specificity, and cost. Almost 400 moths were captured during the experiment. Differences were found in the number of S. frugiperda moths captured in the various trap designs and with the three pheromone lures, and in the number of non‐target moths captured with each lure. The merits of each trap × lure combination are discussed with respect to use in Africa. A nearly equal number of COI‐CS (161) and COI‐RS (158) moths was captured with no differences found in COI marker proportions among traps or lures. However, the diagnostic rice strain marker Tpi was rarely found. Overall, the genetic characterization of the pheromone trap collections indicated a consistent distribution of genetic markers from 2016 to 2017, suggesting a population at or near equilibrium.