Efficacy of diatomaceous earth and methoprene, alone and in combination, against Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) in rough rice

Combination treatments of diatomaceous earth (DE) (Protect-It^®) and the insect growth regulator (IGR) methoprene (Diacon^® II) were evaluated against Rhyzopertha dominica (F.), the lesser grain borer, on stored rough rice. Application rates of DE and methoprene ranged from 0 to 500 ppm and 0 to 1 ppm, respectively, in 25 treatment combinations. Tests were conducted by exposing 20 adults for 2 weeks at 32 °C and 75% relative humidity on single varieties of long-, short-, and medium-grain rough rice, removing adults, and holding the rice for 8 weeks at the same conditions to collect F₁ progeny. In the absence of methoprene, mortality of exposed adults increased as the concentration of DE increased, but even at the highest rate of 500 ppm, mortality was only 57.5±12% and 58.8±9.7% in long and medium-grain rice, respectively, and 26.3±4.7% in short-grain rice. Mortality of R. dominica exposed on short-grain rice was lower than mortality on long-and medium-grain rice at several combinations with 375 and 500 ppm DE. There was also an unexpected increase in adult mortality with the addition of methoprene so that at 1 ppm methoprene and 500 ppm DE, mortality in long-, medium-, and short-grain rice was 77.5±9.0%, 77.5±10.0%, and 58.5±3.0%, respectively. In the absence of methoprene, progeny produced on long- and short-grain rice ranged from 48.0±21.2 to 87.2±9.0, compared to 16.5±5.5 to 33.5±8.6 progeny produced on medium-grain rice. With the inclusion of methoprene there were few progeny produced in any of the treatment combinations, and the overall average was 0.6±0.3. Similarly, with no methoprene the range of insect-damaged kernels (IDK) was 5.2±2.7 to 12.2±3.1%, but with methoprene the overall average was reduced to 1.8±0.2%. While control of R. dominica was somewhat limited with DE, the differences among rice varieties seems to indicate that the specific type and possibly variety of rough rice may influence mortality and reproduction of R. dominica exposed to DE. With methoprene, progeny production was greatly suppressed regardless of DE concentration, but combining DE with methoprene would give some measure of adult control.     

A reliable character for sexing live or dead Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

A new, reliable character is described and illustrated for distinguishing the sexes in adult Rhyzopertha dominica (F.). Both live and dead beetles can be sexed accurately using the method.     

Efficacy of surface applications with diatomaceous earth to control Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) in stored wheat

Commercial formulations of diatomaceous earth (DE) products labeled for use as grain protectants usually specify on the label the depth for using them as a surface treatment, which is often 30.5 cm. An experiment was conducted at two temperatures (27 and 32 °C) and three exposure intervals (7, 10 and 14 d), at a relative humidity of 57–60% to determine if Rhyzopertha dominica (F.), the lesser grain borer, could penetrate a 30.5-cm layer of wheat treated with the labeled rates of three commercial formulations of DE, and, if so, to measure rates of adult survival and progeny production. When R. dominica adults were introduced to this surface layer of 30.5-cm wheat admixed with DE, they were able to penetrate the DE-treated layer and oviposit in the untreated wheat below. Both adult survival and progeny production were significantly lower in wheat with a surface-layer treatment of Dryacide^® (1000 ppm) as compared to Insecto™ (500 ppm), Protect-It^® (400 ppm) or the untreated control. Temperature and exposure interval had no effect on adult survival or progeny production. The vertical displacement patterns of adults were significantly different among DE treatments, but not for temperature or exposure intervals. More R. dominica traveled a greater distance in the untreated control, followed by Insecto™, Protect-It^®, and then Dryacide^®. Results indicate that R. dominica can penetrate a surface layer of DE-treated wheat and reproduce within and below it, but it is possible that pest suppression is dose dependent, or it may depend on a combination of application rate and specific DE formulation.     

Methods for collecting eggs and monitoring egg-hatch and immature development of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

Methods are described for collecting large numbers of R. dominica eggs, assessing the proportion which hatch and rearing first instar larvae to adults. The proportion of eggs which hatched and duration of immature development were estimated at 26 and 30°C, 56% r.h. Mature adults in healthy cultures laid rafts of eggs in between layers of filter papers. The proportion of eggs that hatched was assessed after incubation. The first instar larvae which hatched out of the eggs were reared to adults in individual wells of 96-well polystyrene ELISA plates. They were fed with kibbled wheat while their survival and rate of development was monitored. About half the eggs which were collected shrivelled up without developing. Over 80% of the remaining eggs hatched normally at both temperatures. The mean duration of development from egg to adult through four larval instars was 44 d at 26°C and 34d at 30°C on kibbled wheat. These development times are similar to those on whole wheat reported by other researchers. The methods are much less disruptive or destructive than other methods of monitoring development on flour or whole wheat. Potential applications for these techniques are discussed.     

Evaluation of methoprene alone and in combination with diatomaceous earth to control Rhyzopertha dominica (Coleoptera: Bostrichidae) on stored wheat

A series of experiments were conducted in which different formulations of the insect growth regulator methoprene were evaluated for control of Rhyzopertha dominica (F.), the lesser grain borer, a major internal insect pest of stored wheat. In the first test, application rates of 10-ppm R,S-methoprene (a racemic mixture of the R and S isomers of methoprene) and 1 and 5-ppm S-methoprene (S-isomer only) gave 100% suppression of F₁ adult progeny of R. dominica for 24 weeks. In the second test, adult R. dominica were exposed at 27°C and 32°C, 57% and 75% relative humidity (r.h.) on untreated wheat and wheat treated with 1- to 10-ppm S-methoprene dust. Survival after a 3-week exposure decreased with increasing concentration of dust, and ranged from 69% to 99%, but no F₁ adult progeny were produced in treated wheat. In the final test, concentrations of 0, 0.25, 0.50, 0.75, and 1.0 ppm S-methoprene EC were combined with concentrations of 0, 75, 150, 225, and 300 ppm of the commercial diatomaceous earth (DE) Protect-It^®. Within each methoprene concentration, survival generally decreased with increasing concentration of DE, and was generally greater at 75% than at 57% r.h. Only the wheat treated with 0-ppm methoprene contained an appreciable number of F₁ adults. In summary, both the dust and EC formulations of S-methoprene gave 100% suppression of F₁ adult progeny R. dominica at application rates of 1 ppm, and combination treatments involving reduced rates of methoprene and DE gave effective control of R. dominica.     

The influence of the herbs sage and wormwood on the migration of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) populations

The repellent or attractant effects of adding powdered sage, Salvia officinalis, or wormwood, Artemisia absinthium, to wheat on the migratory behaviour of the lesser grain borer Rhyzopertha dominica was investigated. Under conditions permitting the migration of adult R. dominica, the addition of powdered sage to wheat grain resulted in a lower colonisation of this substrate. It was also found that emigration from grain containing sage was very high, especially in the initial period of grain infestation. This leads to the conclusion that sage is a strong repellent. On the other hand, the addition of pulverised wormwood to wheat did not seem to cause significant mortality or migration activity. Females demonstrated a higher sensitivity to the action of herbal chemicals than males, as confirmed by their higher mortality. This resulted in a change in the sex ratio of the population.     

Survival and reproduction of lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) on flora associated with native habitats in Kansas

Capture of Rhyzopertha dominica in pheromone-baited traps located many kilometers away from grain storage suggests that this species may be exploiting non-grain food sources. To investigate the potential for survival of this species on alternative hosts in the absence of grain, we conducted no-choice feeding assays with twigs and seeds of trees, and seeds of grasses, forbs, and shrubs, commonly encountered on Kansas prairie landscapes where R. dominica has been captured. In addition, R. dominica development and progeny production were assessed on seeds that adults were able to survive on for at least two weeks. Adult survival was poor on grass and forb seeds, although 13.4% of individuals survived on Elymus canadensis, Canada wildrye, seeds after two weeks, compared with 80.3% on Triticum aestivum, wheat, and 0.0% survival in the absence of food. Beetles readily tunneled into twigs from a wide range of tree species, but survival was generally low. A few individuals survived up to four weeks on Gleditsia triacanthos, honey locust. In contrast, bioassays showed that R. dominica could survive and reproduce on damaged seeds (acorns) of six species of oak, but not on undamaged seeds. For example, survival was 95.8% on damaged Quercus muehlenbergii, chinquapin oak. A survey of acorns at the original field collection site showed that the majority of the acorns on the ground had been cracked or bored into by insects and small mammals. Furthermore, we recovered three R. dominica adults from acorns collected in the field and held in sealed containers in the laboratory. These findings suggest that R. dominica populations in Kansas can persist on wild acorns when grain is not available.     

Insecticidal activity of monoterpenes against Rhyzopertha dominica (F.) and Tribolium castaneum (Herbst)

Secondary metabolites play an important role in plant resistance to insects. The monocyclic monoterpenes 1,8-cineole (eucalyptol) and R-(+)-limonene have been considered economically important in this context. Cineole is a component of different species of Eucalyptus spp. and limonene is a constituent of Citrus spp. essential oils. These substances were bioassayed to determine possible fumigant, contact, and ingestion activity against Rhyzopertha dominica (F.), lesser grain borer, and Tribolium castaneum (Herbst), red flour beetle, which are important pests of stored grain. It was possible to show that both compounds are insecticidal against the two insects, mainly in the contact and/or ingestion action.     

The mortality of Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) and Sitophilus oryzae (L.) (Coleoptera: Curculionidae) at moderate temperatures

A method using conductive heating is described for quickly obtaining and maintaining moderate temperatures in grain while minimising grain moisture loss. A model is developed to analyse insect mortality at a range of conditions on log-time logit and log-time probit scales. Values of LT 50, LT 99, and LT 99.9 are presented for adult and immature stages of Sitophilus oryzae in wheat from 42 to 48°C at 12% m.c. and for immature stages of Rhyzopertha dominica in wheat from 45 to 53°C at 9, 12 and 14% m.c. Data show that the heat tolerance of all life cycle stages for both species are different, with a degree of progressive change relative to each other as temperature changes. For R. dominica, the thermal tolerance of life cycle stages also changes relatively with changes in grain moisture content. In general, all stages survive longer at a given temperature as grain moisture increases and the effect of moisture on survival increases as temperature decreases. Values for LT 99.9 for the most heat tolerant stage of R. dominica at 9% m.c. range from 78.22 to 2.49 h between 45 and 53°C. At 12% m.c. they range from 96.81 to 3.36 h, and at 14% m.c. from 114 to 4.14 h. In general, at 12% m.c. the heat tolerance of S. oryzae is about 5°C less than R. dominica with LT 99.9 ranging from 37.36 to 3.71 h between 42 and 48°C. At this level of mortality, immature stage three is generally the most heat tolerant stage of R. dominica, whereas it is stage two for S. oryzae. The results are discussed in relation to practical applications of heat disinfestation and in terms of a general model of mortality. The results are also considered in the context of other available information on the heat treatment of both species.     

Phoretic load of the parasitic mite Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae) affecting mobility and flight take-off of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

Phoresy is the passive transport of an organism on the body of another from a different species, which is a beneficial adaptation for entomophagous arthropods to reach their hosts. Such an interaction occurs between the parasitic mite Acarophenax lacunatus (Prostigmata: Acarophenacidae) and its host, the lesser grain borer Rhyzopertha dominica (Coleoptera: Bostrichidae). Phoresy may have indirect effects in the host species determined by the number of phoretic individuals per carrier (phoretic load) and their distribution on the carrier's body. In the present study, the phoretic load and body distribution of A. lacunatus on adults of R. dominica was assessed. Also the potential interference of the above parameters with host mobility and flight take-off ability were evaluated. Phoretic load and body distribution were assessed by visual inspection under a stereomicroscope, while host mobility was assessed using a computer-automated tracking system. Flight take-off ability was assessed in crowed arenas (containing 50 insects each) at 30 ^°C. The observed mite load ranged from 1 to 11 mites/host, with a peak at around 5 mites/host. Among the host body parts, mites were mainly attached to the thorax (3.70 ± 0.17 mites), where attachment prevailed on the legs and metathorax. Host take-off ability was significantly decreased in the presence of high loads (>7 mites/host) of phoretic mites. Although high phoretic loads (>7 mites/insect) impaired host walking, phoretic loads between 4 and 6 mites/host appeared to favor host walking and consequently the surface dispersal of the parasitic mite.     

Influence of temperature on the functional response of Anisopteromalus calandrae (Hymenoptera: Pteromalidae), a parasitoid of Rhyzopertha dominica (Coleoptera: Bostrichidae)

The functional response of Anisopteromalus calandrae (Howard) parasitizing 4th-instar Rhyzopertha dominica (F.) on wheat was estimated over a range of temperatures and host densities. A functional response equation was used in which a quadratic component that included temperature was substituted for handling time. The instantaneous search rate increased with increasing temperatures. The maximum rate of parasitization was 13 larvae/24 h at 30°C and 35°C. Handling time was lowest at 30°C and highest at 20°C. The ability of A. calandrae to find and parasitize R. dominica over a broad range of temperatures makes it a good candidate for natural control of stored grain pests.     

Uneven application can influence the efficacy of s-methoprene against Rhyzopertha dominica (F.) in wheat

The Juvenile Hormone analogue s-methoprene is used to protect stored grain from pests such as the lesser grain borer, Rhyzopertha dominica (F.). The possibility that uneven application influences s-methoprene efficacy against this species was investigated in the laboratory. Adults of methoprene-susceptible strains were exposed for 14 days to wheat treated at doses of up to 0.6 mg kg⁻¹, or to mixtures of treated and untreated wheat giving equivalent average doses. Adult mortality after exposure to treated wheat was negligible in all cases (≤3.3%) and there was no significant effect of either average dose or evenness of application. In contrast, the number of adult progeny depended on both the average dose and evenness of application. Average doses of 0.3 and 0.6 mg kg⁻¹ reduced the number of live F₁ adults by 99–100% relative to the untreated wheat and no effect of evenness of application was detected. At lower doses, however, efficacy tended to decrease with increasing unevenness of application. When adults from the parental generation were transferred to untreated wheat for another 14 days neither the average dose nor evenness of application in the wheat from which they came had any significant effect on reproduction of these adults. This study demonstrates that uneven application can reduce the efficacy of s-methoprene against R. dominica, but that this is unlikely to influence the performance of s-methoprene against susceptible populations at target doses likely to be used in practice (e.g. 0.6 mg kg⁻¹ in Australia). However, the possibility that uneven application leads to underdosing and selects for resistance should be investigated.     

Efficacy of diflubenzuron plus methoprene against Sitophilus oryzae and Rhyzopertha dominica in stored sorghum

The efficacy of diflubenzuron (1 mg kg⁻¹)+methoprene (1 mg kg⁻¹) against Sitophilus oryzae (L.) and Rhyzopertha dominica (F.) in sorghum was evaluated in a silo-scale trial in southeast Queensland, Australia. Sorghum is normally protected from a wide range of insects by mixtures of grain protectants. The chitin synthesis inhibitor diflubenzuron was evaluated as a potential new protectant for S. oryzae in combination with the juvenile hormone analogue methoprene, which is already registered for control of R. dominica. Sorghum (ca 200 t) was treated after harvest in 2000 and assessed for treatment efficacy and residue decline during 6.5 months storage. The reproductive capacity of S. oryzae and R. dominica was greatly reduced in bioassays of treated sorghum throughout the trial, and efficacy remained relatively stable during the trial. An initial exposure of S. oryzae adults to treated sorghum for 2 weeks reduced F₁ progeny production of all strains by 80.8-98.8%, but a second exposure of 4 weeks reduced F₁ progeny production by 98.5-100%. In addition, the reproductive capacity of any S. oryzae progeny produced was greatly reduced. Exposure of R. dominica adults to treated sorghum for 2 weeks reduced F₁ progeny production of all strains by 99.6-100%, including a methoprene-resistant strain. The results indicate that S. oryzae or R. dominica adults invading sorghum treated with diflubenzuron (1 mg kg⁻¹)+methoprene (1 mg kg⁻¹) would be incapable of producing sustainable populations.     

Hull characteristics as related to susceptibility of different varieties of rough rice to Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

Rhyzopertha dominica (F.), an important pest of stored grains, causes economic damage to rough rice through physical damage to the kernel, resulting in reductions in grain quality. In this test, 28 varieties of commercial rough rice (10 long grain, 11 medium grain, and 7 short grain) were examined for solid, split and cracked hulls, hull thickness, and adult emergence from neonate R. dominica introduced on each individual variety. The percentage of solid hulls ranged from 55.5% on Koshihikari variety to 92.8% on Akita variety, and the percentages of cracked and split hulls were correlated with increased susceptibility. The Dobie index for progeny production showed Wells, Jupiter, and Pirogue varieties as the most tolerant to R. dominica, while Rico and Francis were the most susceptible. The hull thickness of rough rice varied among varieties, but the tolerant varieties appeared to have thicker hulls than the susceptible varieties. There was no difference among rice types (long-, medium-, or short grain) regarding tolerance or susceptibility to R. dominica. Results show that the characteristics of the rough rice hull are important for conferring susceptibility of individual varieties to R. dominica.     

Detection and quantification of the insect pest Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) in rice by qPCR

The early detection of insects during grain storage and processing remains a major issue for the cereal industry, especially when immature stages are hidden inside the grain kernels. For this reason, we developed a qPCR method to detect and quantify one of the main pests of stored products in rice: the coleopteran internal feeder Rhyzopertha dominica. For that purpose, a specific primer set was designed to amplify artificial infestations of this pest in rice. Then, using a regression model, a standard curve was generated that correlated individuals to adult equivalent DNA quantity (inverse of the C_t value). Results revealed that the designed primer set was specific for R. dominica when tested against the other 4 common internal feeders in grain. The technique showed to be accurated (DNA was detected in more than 73% of the samples) and sensitive to insect presence (i.e. from 0.02 adults, 0.1 3rd instar to pupae or 13 egg to 2nd instar detectable per kg of rice). Moreover, the detection of R. dominica was strongly associated with a given infestation size: DNA quantity increased along with the size of the population. The use of the described qPCR protocol in grain and milling factories may enhace the critical detection and quantification of R. dominica populations in raw materials and processed food.     

Susceptibility of the gelatinized wheat product bulgur to Sitophilus zeamais Motsch. and Rhyzopertha dominica (F.)

The development of Sitophilus zeamais and Rhyzopertha dominica was studied at relative humidities of 71, 58 and 43% in bulgur and wheat. Male and female adult weevils, introduced into 6 bulgur and 6 wheat media of different particle sizes, were allowed to oviposit for 7 days. Thirty R. dominica eggs were placed in each replicate of a similar set of bulgur and wheat media.

S. zeamais produced more adult progeny in whole kernels of bulgur and of wheat and in the larger-sized particles, and a high percentage of R. dominica larvae developed into adults in these media. S. zeamais progeny produced and R. dominica larvae surviving were fewer, and developmental periods for both species longer, the smaller the particles and the lower the relative humidity. Males and females collected from bulgur and wheat media of the same particle size tended to weigh the same; but the smaller the particle size, the less the weights.     

Sexing live adult Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae)

A recently published method of sexing live adult Rhyzopertha dominica (F.) using a punctuate groove on the fifth abdominal sternite of the males was found unsatisfactory for the strains from Queensland, Australia. Detailed assessment was made of the squeezing and colour methods of sexing live adult beetles. Squeezed virgin females subsequently mated to untreated males had a reduced fecundity. Untreated females mated to squeezed males also produced fewer eggs than the control although the mating ability of squeezed males was not significantly affected by the treatment. Squeezing significantly reduced the life expectancy but not of males. One observer caused more damage than the other. In colour sexing a greater accuracy was achieved by avoiding difficult choices. Egg production of females with strong colour marking was not significantly lower than that of unselected females.     

Proteomic assessment of resistance to the fumigant phosphine in the lesser grain borer, Rhyzopertha dominica (F.)

A previous publication compared a strain of the stored-grain pest beetle, Rhyzopertha dominica, resistant to the fumigant phosphine, with a susceptible strain using two-dimensional polyacrylamide gel electrophoresis. Certain proteins appeared to differ and the authors proposed that arginine kinase in particular might be used as a marker for the rapid monitoring of resistance in this species and other pests of stored grain. Here an expanded set of susceptible and resistant strains is surveyed using the two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) system. Of hundreds of spots only two showed a significant difference between the resistant and susceptible strains and the magnitudes of those differences were less than three-fold. Arginine kinase spots were identified but they did not differ significantly. The proposal that certain abundant proteins of R. dominica might be used as markers of phosphine resistance was not supported by the present study.     

Relative effectiveness of topical, filter paper and grain applications of deltamethrin, and associated behaviour of Rhyzopertha dominica (F.) strains

The susceptibility of stored grain pests to insecticides may be assessed by different types of bioassay application methods such as topical, impregnated filter papers and grain treatments. Pests may behave differently when in contact with pesticides and these changes in behaviour may contribute to their tolerance of pesticide. To compare the three application methods and determine how Rhyzopertha dominica (F.) behaved on contaminated filter papers and treated grain, experiments were carried out on four strains of this species. The strains were of known susceptibility to deltamethrin; three were from Brazil (BR4, BR6 and BR7) and one from the United Kingdom (UK1). BR4 and UK1 were susceptible strains, and BR6 and BR7 were resistant strains. Behavioural responses of the pest to deltamethrin were compared between resistant and susceptible strains by measuring the walking response of individuals on deltamethrin impregnated filter papers and the repellent effect of treated wheat grain. The body weights of strains were compared. The results showed resistance ratios of 21, 175 and 17 times between the most susceptible and the most resistant strains, following topical, filter paper and grain applications, respectively. The most resistant strain walked less on contaminated filter papers while the most susceptible strain did not change its walking pattern. Deltamethrin showed no repellent effect, neither on impregnated filter papers or in treated wheat grain. The body weights differed among the strains but were not associated with the level of tolerance to deltamethrin.     

Efficacy of protectants against Callosobruchus phaseoli (Gyll.) and C. maculatus (F.) (Coleoptera: Bruchidae) in mungbeans

The organophosphorus (OP) compounds chlorpyrifos-methyl, fenitrothion, methacrifos and pirimiphos-methyl, synthetic pyrethroids (SP) deltamethrin and permethrin, and the insect growth regulator (IGR) fenoxycarb were evaluated as protectants against Callosobruchus phaseoli (Gyll.) and C. maculatus (F.) in mungbeans (Vigna radiata (L.) Wilczek). Treated mungbeans were sampled for efficacy testing using C. phaseoli after various periods of storage in hot and humid conditions (30°C, 70% r.h.). Minimum effective application rates were determined for different periods of protection. During storage the OP compounds (chlorpyrifos-methyl, fenitrothion, methacrifos and pirimiphos-methyl) lost activity against C. phaseoli much faster than the SPs (deltamethrin and permethrin) and the IGR (fenoxycarb). Tests were carried out on mungbeans which were infested with C. phaseoli before treatment with pirimiphos-methyl, fenoxycarb or deltamethrin. These tests showed that immature insects in the treated beans were protected from insecticides. Finally, freshly-treated mungbeans were exposed to C. maculatus for comparison with the results for C. phaseoli. The four OP compounds, two SPs and the IGR tended to have similar potency against C. maculatus and C. phaseoli.