Potential of the neonicotinoid imidacloprid and the oxadiazine indoxacarb for controlling five coleopteran pests of stored grain

Abstract The potential for using imidacloprid (a neonicotinoid) and indoxacarb (an oxadiazine) as grain protectants was investigated in bioassays against resistant strains of five stored grain beetles. The species investigated were Rhyzopertha dominica (F.) (the lesser grain borer), Sitophilus oryzae (L.) (the rice weevil), Tribolium castaneum (Herbst) (the rust‐red flour beetle), Oryzaephilus surinamensis (L.) (the saw tooth flour beetle), and Cryptolestes ferrugineus (Stephens) (the flat grain beetle). Each of these species has developed resistance to one or more protectants, including organophosphorus insecticides, synthetic pyrethroids and the juvenile hormone analogue methoprene. Mortality and reproduction after a 2‐week exposure of adults to treated wheat depended on species, dose and insecticide. Imidacloprid had no effect on S. oryzae at any dose, but none of the other species produced any live progeny at 10 mg/kg. Indoxacarb had no effect on T. castaneum at any dose, but none of the other species produced any live progeny at 5 mg/kg. The results show that although both imidacloprid and indoxacarb can control at least four of the five key pests tested at doses comparable to those used for organophosphorus protectants, more potent neonicotinoid or oxadiazine insecticides would be needed than either of these to provide broad spectrum protection of stored grain.     

Effect of five diatomaceous earth formulations against Tribofium castaneum （Coleoptera： Tenebrionidae）, Oryzaephflus surinamensis （Coleoptera： Silvanidae） and Rhyzopertha dominica （Coleoptera： Bostrychidae）

Laboratory bioassays were conducted to determine the effect of food source on the survival of Tribolium castaneum Herbst, Oryzaephilus surinamensis L. and Rhyzopertha dominica F., after exposure to five diatomaceous earth （DE） formulations： Protect-It, Insecto, Perma-GuardTM, Dryacide and SilicoSec. Adults of these species were exposed to DEs at the rate of 0.5 mg/cm^2 for 1 day on filter paper inside plastic Petri dishes. After exposure, the initial mortality was counted and live individuals of the three species were held for a week in glass vials containing 50 mg wheat flour, rice and whole wheat, respectively. In the second experiment, after 1 day exposure to DEs, beetles were transferred to Petri dishes without food and held for a week to determine if the presence of food source would decrease the mortality of beetles. Experiments were carried out at 27℃ and 55% RH in the dark. The initial mortality in both of the experiments reached 100% for the three species exposed to Protect-It and in the case ofR. dominica and O. surinamensis exposed to Dryacide. In contrast, low level of mortality （〈 10%） was observed for T. castaneum exposed to Perma-GuardTM and Insecto. The mortality after the post-treatment period on food was decreased for the three species exposed to Perma-GuardTM and in the case of T. castaneum and R. dominica exposed to Insecto and SilicoSec. Adults of O. surinamensis were the most susceptible followed by R. dominica and 100% adult mortality was obtained, whereas T. castaneum were the least susceptible beetles to DEs. Protect-It and Dryacide were the most efficient DE formulations and can be used effectively in a stored grain integrated pest management program.     

Effects of bean and wheat α-amylase inhibitors on α-amylase activity and growth of stored product insect pests

Insect α-amylase inhibiting and/or growth inhibiting activities of proteinaceous inhibitors from red kidney bean (Phaseolus vulgaris) and hard red winter wheat (Triticum aestivum) were examined. The bean inhibitor was most effectivein vitro against α-amylases from the red flour beetle (Tribolium castaneum) and the confused flour beetle (T. confusum), followed by those from the rice weevil (Sitophilus oryzae) and yellow mealworm (Tenebrio molitor). The insect enzymes were from two- to 50-fold more susceptible than human salivary α-amylase. When the inhibitors were added at a 1% level to a wheat flour plus germ diet, the growth of red flour beetle larvae was slowed relative to that of the control group of larvae, with the bean inhibitor being more effective than the wheat inhibitor. Development of both the red flour beetle and flat grain beetle (Cryptolestes pusillus) was delayed by 1% bean inhibitor, but development of the sawtoothed grain beetle (Oryzaephilus surinamensis) and lesser grain borer (Rhyzopertha dominica) was not affected by either the bean or wheat inhibitor at the 1% level. Rice weevil adults fed a diet containing 1% bean or wheat inhibitor exhibited more mortality than weevils fed the control diet. When the wheat amylase inhibitor was combined with a cysteine protease inhibitor, E-64, and fed to red flour beetle larvae, a reduction in the growth rate and an increase in the time required for adult eclosion occurred relative to larvae fed either of the inhibitors separately. The bean inhibitor was just as effective alone as when it was combined with the protease inhibitor. These results demonstrate that plant inhibitors of insect digestive enzymes act as growth inhibitors of insects and possibly as plant defense proteins, and open the way to the use of the genes of these inhibitors for genetically improving the resistance of cereals to storage pests. Cooperative investigation between the Agricultural Research Service, the University of California, San Diego, and the Kansas Agricultural Experiment Station (Contribution no. 94-416-J). Supported in part by a grant from the Ministry of Education and Science, Spain-Fulbright Program to J.J.P. Mention of a proprietary product does not constitute a recommendation or endorsement by the USDA. The USDA is an equal opportunity/affirmative action employer and all agency services are available without discrimination.     

Snowdrop and wheatgerm lectins and avidin as antimetabolites for the control of sugarcane whitegrubs

Snowdrop and wheatgerm lectins were found to be insecticidal and growth inhibiting dietary proteins for larvae of the sugarcane whitegrub Antitrogus parvulus. At concentrations as low as 0.5 mg of snowdrop lectin per gram of semi-artificial diet, growth was inhibited by 21 days of feeding and significant mortality was apparent by 28 days. Wheatgerm lectin was active at similar concentrations, although expression of the effects was slower. Avidin was found to be a growth inhibiting dietary protein for larvae of Antitrogus consanguineus. At levels as low as 0.01 mg g^-1 of diet, growth was inhibited by 28 days of feeding. Avidin caused no significant mortality after 35 days of feeding. Snowdrop and wheatgerm lectins and avidin are insect growth-inhibiting proteins whose genes potentially could be manipulated into sugarcane and improve host-plant resistance to whitegrubs.     

Aerosol distribution and efficacy in a commercial food warehouse

A series of field trials were conducted in a commercial food storage facility to evaluate exposure of stored‐product insects to aerosol formulations of synergized pyrethrins and the insect growth regulator methoprene. When adults of Tribolium castaneum (Herbst), the red flour beetle, and Tribolium confusum (Jacqueline DuVal), the confused flour beetle were exposed with and without a food source to synergized pyrethrin aerosol, there was no difference in adult mortality with respect to availability of food at either 7 or 14 days after exposure (P≥ 0.05). However, mortality was lower in T. confusum (40.4% and 79.3% with flour at 7 and 14 days, 38.9% and 84.8% without flour at 7 and 14 days) compared to T. castaneum (96.5% and 99.8% with flour at 7 and 14 days, 91.0% and 98.7% without flour at 7 and 14 days). Few late‐stage larvae and pupae of either species exposed to the pyrethrin aerosol emerged as adults. In tests with methoprene aerosol, adult emergence of exposed 3‐and 4‐week‐old larvae of T. confusum was less than 2%. Only 0.3% of 4‐week‐old larvae of T. castaneum exposed in open and obstructed areas emerged as adults. Emergence of adults from eggs of Plodia interpunctella (Hübner), the Indianmeal moth, embedded in culture media and exposed to the methoprene aerosol was 13.2%± 3.5%. Results show that the aerosols evaluated in our study could give effective control of some of the major stored‐product insect pests in commercial food storage facilities, and may offer an alternative to fumigation.     

Fumigant toxicity of essential oil of Mugwort (Artemisia vulgaris L.) against three major stored product beetles

Application of plants essential oil for the evaluation of their fumigant toxicity and insecticidal properties is the goal of many researches. In this study, aerial parts of Artemisia vulgaris L. were subjected to hydrodistillation using a Clevenger-type apparatus, and the chemical composition of the volatile oils was studied by gas chromatography–mass spectrometry. Alpha-Pinene (23.56) was the main component of the essential oil. Insecticidal activity of the oil was evaluated against Tribolium castaneum (Herbst), Callosobruchus maculatus (F.) and Rhizopertha dominica (F.) after 24, 48 and 72 h. After 24-h exposure time, C. maculatus was more susceptible (LC₅₀ = 52.47 μl/l air) and T. castaneum was more tolerant (LC₅₀ = 279.86 μl/l air) than other species. LT₅₀ values were indicated using highest concentration of LC₅₀ tests for three species. In general, mortality increased as the doses of essential oil and exposure time increased. These results proposed that A. vulgaris oil might have potential as a control agent against T. castaneum, R. dominica and especially C. maculates in storages.     

Using different amounts of vacuum and acrolein in various exposure periods to control two stored products insects

In this research, adults of Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) were exposed to various air pressures solely (first phase) and air pressures together with acrolein (second phase) in various durations and mortality rates were determined. Complete mortality of T. castaneum was obtained at 1, 3 and 6 h after exposure to 10, 30 and 50 mmHg, respectively. R. dominica showed 96.25% and 60% mortality during 3 and 10 h exposed to 10 and 30 mmHg, respectively. Percentage mortality of the insects in second phase was significantly higher than the rate in first phase. T. castaneum was more susceptible than R. dominica in the both phases.     

Identification of microsatellite markers in the red flour beetle,Tribolium castaneum

We isolated and characterized microsatellite markers for the red flour beetle Tribolium castaneum, an important model species for studies in various areas of evolutionary biology and ecology. A microsatellite‐enriched genomic library was constructed and screened with single stranded oligonucleotide probes [(CCT)₁₇, (AAT)₁₇ and (CAG)₁₇]. Forty‐five primer pairs were designed of which 19 pairs produced successful amplification. Polymorphism screening involved beetles from five beetle strains and revealed 15 polymorphic and four monomorphic markers. The development of polymorphic microsatellite markers will facilitate future ecological and genetic studies involving T. castaneum beetles.     

Differential pathogenicity of two entomopathogenic bacteria,Photorhabdus temperata subsp. temperata and Xenorhabdus nematophila against the red flour beetle,Tribolium castaneum

Two entomopathogenic bacteria, Photorhabdus temperata subsp. temperata (Ptt) and Xenorhabdus nematophila (Xn), are symbiotically associated with the nematodes, Heterorhabdis megidis and Steinernema carpocapsae, respectively. There is little information on natural host ranges of the nematodes, but a significant difference in pathogenicity was observed between these two bacteria against the red flour beetle, Tribolium castaneum, in which Ptt exhibited more than six times higher pathogenicity than Xn. The pathogenic difference was not due to their inhibitory effect on phospholipase A₂ activity that is required for expression of immune response of T. castaneum. The culture broths of both bacterial species had insecticidal activities when injected into the hemocoel. When the bacterial culture broths were fractionated into aqueous and organic extracts, most insecticidal activity remained in the aqueous extracts. The aqueous extracts of two bacteria contained proteins which showed different profiles.     

Response of Tribolium castaneum and Rhyzopertha dominica to various resources,near and far from grain storage

Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.) are common cosmopolitan pests of stored grain and grain products. We evaluated the relative attraction of T. castaneum and R. dominica to wheat, sorghum and cotton seeds in the field, near grain storage facilities and well away from storages in southern and central Queensland using multiple trapping techniques. The results show that T. castaneum is more strongly attracted to linted cotton seed relative to wheat, whereas R. dominica did not respond to cotton seed at all and was attracted only to wheat. Significantly more adults of T. castaneum (10–15 times) were attracted to traps placed on the ground, near grain storage, than to equivalent traps that were suspended (1.5 m above the ground) nearby. These results suggest that Tribolium beetles detect and respond to resources towards the end of their dispersal flight, after which they localize resources while walking. By contrast R. dominica was captured only in suspended traps, which suggests they fly directly onto resources as they localize them. The ability of both species to colonize and reproduce in isolated resource patches within the relatively short time of 1 month is illustrated by the returns from the traps deployed in the field (at least 1 km from the nearest stored grain) even though they caught only a few beetles. The results presented here provide novel insights about the resource location behaviours of both T. castaneum and R. dominica. In particular, the relationship of T. castaneum with non‐cereal resources that are not conventionally associated with this species suggests an emphasis on these other resources in investigating the resource location behaviour of these beetles. This new perspective on the ecology of T. castaneum highlights the potential role of non‐cereal resources (such as the lint on cotton seed) in the spread of grain pest infestations.     

Low temperature shock and chill-coma consequences for the red flour beetle (Tribolium castaneum) and the rice weevil (Sitophilus oryzae)

Insects face several (environmental) abiotic stressors, including low temperature, which cause the failure of neuromuscular function. Such exposure leads insects toa reversible comatose state termed chill-coma, but the consequences of this state for the organism biology were little explored. Here, the consequences of the chill-coma phase were investigated in two of the main stored product pest species – the red flour beetle Tribolium castaneum (larvae and adults) and the rice weevil Sitophilus oryzae (adults). For this purpose, a series of low-temperature shocks were used to estimate the chill-coma recovery time (CCRT), survival, nutrition and weight gain/growth of T. castaneum (larvae and adults) and S. oryzae, as well as the development of T. castaneum life stages. The relatively long CCRT was characteristic of beetle larvae, at different low-temperature shocks, and CCRT increased with decreasing temperatures and increasing exposure intervals for both pest species. The survival was little affected by the low-temperature shocks applied, but such shocks affected insect feeding and growth. Tribolium castaneum larvae was more sensitive than adults of both insect species. Moreover, the relative consumption and weight gain of S. oryzae adults were lower than those of T. castaneum adults and mainly larvae, while feeding deterrence was not affected by low temperature shocks, unlike food conversion efficiency. Low-temperature shocks, even under short duration at some temperatures, significantly delayed development. The lower the temperature and the higher the exposure period, the more delayed the development. Thus, the physiological costs of chill-coma are translated into life-history consequences, with potential implications for the management of this insect pest species in stored products and even more so on red flour beetles and rice weevils.     

Analysis of repetitive DNA distribution patterns in the <Emphasis Type="Italic">Tribolium castaneum</Emphasis> genome

Background  

Insect genomes vary widely in size, a large fraction of which is often devoted to repetitive DNA. Re-association kinetics indicate that up to 42% of the genome of the red flour beetle, Tribolium castaneum, is repetitive. Analysis of the abundance and distribution of repetitive DNA in the recently sequenced genome of T. castaneum is important for understanding the structure and function of its genome.     

Parasitism by the mite Acarophenax lacunatus on beetle pests of stored products

Recent reports of the biocontrolpotential of the mite species Acarophenaxlacunatus (Cross & Krantz) (Prostigmata:Acarophenacidae) and the lack of biologicalstudies on this regulatory agent led to thepresent study carried out under laboratoryconditions. The objective of the investigationwas to assess the host range of A. lacunatus,so far only reported as egg parasite ofRhyzopertha dominica (F.) (Coleoptera:Bostrichidae). Four Coleoptera species ofstored cereals were used: R. dominica,Tribolium castaneum (Herbst) (Tenebrionidae),Cryptolestes ferrugineus (Stephens)(Laemophloeidae) and Oryzaephilus surinamensis (L.) (Cucujidae). The highest rates of eggparasitism were observed on R. dominica and T.castaneum, leading to a significant decrease ofpopulations of both species and reduced wheatweight loss. A. lacunatus was also able toparasitize eggs of C. ferrugineus, but not ofO. surinamensis. These results indicate abroader host range of A. lacunatus thaninitially suspected and also strengthen itspossibility of use in integrated pestmanagement programs in storage environments.     

Population structural changes in single species of flour beetles with ozone as an environmental factor

1. Although a 5 hour monthly exposure for 16 months to 45.0 ppm O₃ at 0.3l/min. reduced the numbers of individuals in single-species populations of T. confusum and T. castaneum, it did not result in elimination or a significant decrease in numbers which would be useful in a control program for these flour beetle species.
2. During the first 5 months of culture, ozone had a stimulating effect on productivity of T. confusum; a similar effect was not noted for T. castaneum.
3. Control of flour beetles by ozone in stored-products seems unlikely; however, ozone in an integrated control program with ionizing radiation might prove beneficial.
4. Biological interpretations are offered to explain the responses of these species to ozone exposures.
5. Further research is warranted to define ozone modifications in pest control programs and on stored-products per se.

     

Diagnostic Molecular Markers for Phosphine Resistance in U.S. Populations of Tribolium castaneum and Rhyzopertha dominica

Stored product beetles that are resistant to the fumigant pesticide phosphine (hydrogen phosphide) gas have been reported for more than 40 years in many places worldwide. Traditionally, determination of phosphine resistance in stored product beetles is based on a discriminating dose bioassay that can take up to two weeks to evaluate. We developed a diagnostic cleaved amplified polymorphic sequence method, CAPS, to detect individuals with alleles for strong resistance to phosphine in populations of the red flour beetle, Tribolium castaneum, and the lesser grain borer, Rhyzopertha dominica, according to a single nucleotide mutation in the dihydrolipoamide dehydrogenase (DLD) gene. We initially isolated and sequenced the DLD genes from susceptible and strongly resistant populations of both species. The corresponding amino acid sequences were then deduced. A single amino acid mutation in DLD in populations of T. castaneum and R. dominica with strong resistance was identified as P45S in T. castaneum and P49S in R. dominica, both collected from northern Oklahoma, USA. PCR products containing these mutations were digested by the restriction enzymes MboI and BstNI, which revealed presence or absence, respectively of the resistant (R) allele and allowed inference of genotypes with that allele. Seven populations of T. castaneum from Kansas were subjected to discriminating dose bioassays for the weak and strong resistance phenotypes. Application of CAPS to these seven populations confirmed the R allele was in high frequency in the strongly resistant populations, and was absent or at a lower frequency in populations with weak resistance, which suggests that these populations with a low frequency of the R allele have the potential for selection of the strong resistance phenotype. CAPS markers for strong phosphine resistance will help to detect and confirm resistant beetles and can facilitate resistance management actions against a given pest population.     

UV resonance Raman study of streptavidin binding of biotin and 2-iminobiotin: comparison with avidin

UV resonance Raman (UVRR) spectroscopy is used to study the binding of biotin and 2-iminobiotin by streptavidin, and the results are compared to those previously obtained from the avidin-biotin complex and new data from the avidin-2-iminobiotin complex. UVRR difference spectroscopy using 244-nm excitation reveals changes to the tyrosine (Tyr) and tryptophan (Trp) residues of both proteins upon complex formation. Avidin has four Trp and only one Tyr residue, while streptavidin has eight Trp and six Tyr residues. The spectral changes observed in streptavidin upon the addition of biotin are similar to those observed for avidin. However, the intensity enhancements observed for the streptavidin Trp Raman bands are less than those observed with avidin. The changes observed in the streptavidin Tyr bands are similar to those observed for avidin and are assigned exclusively to the binding site Tyr 43 residue. The Trp and Tyr band changes are due to the exclusion of water and addition of biotin, resulting in a more hydrophobic environment for the binding site residues. The addition of 2-iminobiotin results in spectral changes to both the streptavidin and avidin Trp bands that are very similar to those observed upon the addition of biotin in each protein. The changes to the Tyr bands are very different than those observed with the addition of biotin, and similar spectral changes are observed in both streptavidin and avidin. This is attributable to hydrogen bond changes to the binding site Tyr residue in each protein, and the similar Tyr difference features in both proteins supports the exclusive assignment of the streptavidin Tyr difference features to the binding site Tyr 43.     

Mixing of Isaria fumosorosea with enhanced diatomaceous earth and bitterbarkomycin for control of Rhyzopertha dominica

The virulence of Isaria fumosorosea alone and integrated with an enhanced formulation of diatomaceous earth (diatomaceous earth + bitterbarkomycin, DEBBM) was tested against the lesser grain borer Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) under laboratory conditions on stored wheat. The fungus was applied to wheat grains at various concentrations of 3.60 × 10⁶, 3.60 × 10⁸, and 3.60 × 10¹⁰ conidia/kg alone, or in combination with 15 and 30 ppm of DEBBM. R. dominica was held at 20, 25 and 30°C and two RH regimes (56 and 75%) and mortality was registered after 5, 10 and 15 d. Generally, I. fumosorosea was most effective at 25°C caused the highest mortality of R. dominica at 56% RH. The increase of temperature, concentration and exposure interval increased the insecticidal effect of DEBBM at low RH level. The DE alone suppressed the progeny emergence at higher dose rate as compared I. fumosorosea alone, but their simultaneous use further reduced the progeny production of R. dominica. The integrated use of both tested bio control agents synergized the effect of each other and caused the highest mortality at 25°C and 56% RH. The results clearly demonstrated that I. fumosorosea and new enhanced DEBBM can be integrated to be an effective control measure for R. dominica in stored wheat.     

Cold tolerance in flour beetle species differing in body size and selection temperature

Cold tolerance is an important trait directly related to survival and hence fitness. In the present study, the link is addressed between cold tolerance and body size, which is associated with many key fitness traits, at both the intra‐ and interspecific levels. Specifically, chill coma recovery time, as a metric of cold tolerance, is examined in five related flour beetle species (four of them belonging to the genus Tribolium), two additional Tribolium castaneum Herbst populations selected for higher temperatures, and a mutant showing reduced body size. Recovery times are negatively correlated with the species average body size but not within each species. Females usually recover faster than males, although this difference is significant in only a single species, and is unrelated to body size. Repeating the experimental procedure with the same individuals, after 2 days in isolation with a limited amount of food, results in longer recovery times. Therefore, even if cold acclimation takes place, its influence appears to be diminished by the deleterious effects associated with the experimental procedure. Hence, the findings provide evidence for an association between body size and cold tolerance in the genus Tribolium, with larger species recovering faster from chill than smaller species. By contrast, the smalleyed flour beetle Palorus ratzeburgii Wissmann does not follow this pattern. Additionally, a population of T. castaneum selected for the highest temperature takes longer to recover from chill coma, indicating a trade‐off between cold and heat adaptations and not to a cross‐protection effect, as sometimes demonstrated.