Toxicity of ethyl formate to adult Sitophilus oryzae (L.), Tribolium castaneum (herbst) and Rhyzopertha dominica (F.)

Fumigations were conducted using a continuous flow-through laboratory process to maintain constant concentrations of ethyl formate and low levels (<0.8%) of respiratory carbon dioxide. The procedure minimised the effects of sorption by exposing test insects without media and minimised the effect of carbon dioxide by use of continuous flow. The concentration×time (Ct) products of ethyl formate for adult Sitophilus oryzae, Tribolium castaneum and Rhyzopertha dominica at 25 °C and 70% relative humidity for the 6 h exposure were, respectively: (1) LD₅₀ 107.8, 108.8 and 72.8 mg h L⁻¹ and (2) LD_99.5 207.4, 167.1 and 122.2 mg h L⁻¹. Endpoint mortality was reached within 24 h of initial exposure.     

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.     

Effects of starvation period on the locomotory response of Rhyzopertha dominica (F.)

In the context of developing a behavioural bioassay, the effects of 1-5 days starvation on the locomotory responses of Rhyzopertha dominica were investigated. Movements in an experimental arena with airflow were videoed and analysed using computerised tracking and motion analysis software. Beetle velocity and angular velocity (turning) were influenced adversely following 3 or 4 days of starvation, resulting in a lower number of beetles arriving at food. The highest number of beetles arriving at food was when they were starved for 1 day. Gender had a statistically significant effect on velocity; on average males walked 37% faster than females and there were significant decreases in the velocity of females from the fourth day and male velocity from the fifth day of starvation. Starvation significantly reduced angular velocity of beetles although the effect of gender was not statistically significant. There was a decline in angular velocity following the first day of starvation, although the negative effect of starvation on turning is not as clear-cut as on velocity, with a more gradual decline with increasing starvation. For undertaking behavioural bioassays with R. dominica, starvation beyond 1 day is unlikely to improve responsiveness.     

Screening of Chinese medicinal herbs for bioactivity against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst)

Extracts of 40 species of Chinese medicinal herb from 32 different botanical families were screened for contact, fumigant and feeding-deterrent activities against the two stored-grain insects Sitophilus zeamais and Tribolium castaneum. Thirty Chinese medicinal herbs exhibited insecticidal or feeding-deterrent activities against the two species of insects. Extracts of Artemisia argyi, Dictamnus dasycarpus, Evodia rutaecarpa, Litsea cubeba, Narcissus tazetta var. chinensis, Polygonum aviculare, Rhododendron molle, Sophora flavescens, Stemona sessilifolia, Tripterygium wilfordii, and Torreya grandis were most active.     

Respiration of Rhyzopertha dominica (F.) at reduced oxygen concentrations

Developmental stages of Rhyzopertha dominica were exposed to atmospheres containing 1%, 2%, 3%, 5%, 10% or 15% oxygen (O₂) in nitrogen at 30°C and 70% r.h. Respiration rates were determined with a gas chromatograph. The O₂ intake and carbon dioxide (CO₂) output by insects were expressed in μl/insect h or μl/mg h. Respiration of eggs, young and old larvae, pupae, and adults at normal atmospheric air were at rates of 0.0029, 0.41, 2.52, 0.82, and 2.86 μl CO₂/insect h, respectively. Respiration rates of the same stages in terms of insect weight were 0.14, 4.83, 1.98, 0.64 and, 2.58 μl CO₂/mg h, respectively. At reduced O₂ levels respiration rates of eggs, larvae and pupae were proportional to the O₂ levels. Adult respiration rates were high at 3% and 5% O₂ levels almost reaching that of normal atmospheric air, and were 2.56 and 2.85 μl CO₂/insect h, respectively. In adults, respiration quotient values for the same O₂ levels were higher than at normal atmospheric O₂ and were 1.5 and 1.02, respectively.Respiration of adults in normal air between 20°C and 35°C increased with temperature and gas values varied between 0.89 and 6.82 μl CO₂/insect h, respectively, or 0.93 and 5.63 μl O₂/insect h, respectively.     

Fecundity of wild Tribolium castaneum (Herbst) from southeast Queensland, Australia

Fecundity was investigated in wild Tribolium castaneum (Herbst) from southeast Queensland, Australia. Although the fecundity of T. castaneum has been investigated in laboratory studies, the reproductive potential of wild females may be less than that of females reared and tested under favourable laboratory conditions. Adult T. castaneum were trapped using corrugated cardboard traps baited with whole wheat flour on four occasions during May-July 1997 in a grain-growing area of southeast Queensland. After each trapping period, fecundity of individual wild females was assessed on whole wheat flour at 30°C and 55% erh. In addition, F₁ adults were pooled, a sample was taken and fecundity of individual females was assessed similarly. Almost 90% of the adults were trapped on four of the 10 grain storage sites, and these sites yielded females on all four trapping occasions. There was no systematic effect of trapping period on fecundity of wild females, and in three of these four sites there was no effect at all. There were no significant differences between numbers of progeny produced by F₁ females derived from different trapping periods, so the data were pooled for comparison with the results from wild females. Generally, fecundity of wild females was high relative to that of F₁ females which had been reared under uniformly favourable conditions in the laboratory. Despite the trapping occurring during some of the coldest months of the year, females were clearly active and capable of reproducing, demonstrating the importance of maintaining high levels of grain hygiene throughout the year.     

Calorimetric evaluation of responses of Sitophilus oryzae and Tribolium confusum to elevated temperatures and controlled atmospheres

Metabolic heat rates, determined by microcalorimetry, were used to measure the effect of controlled atmospheres (CAs) and elevated temperatures on the stored-product insects Sitophilus oryzae (rice weevil) and Tribolium confusum (confused flour beetle). Results for larval and adult stages in air, and in a range of low O₂ and/or high CO₂ CAs, at temperatures from 15 to 45 °C, showed the general effectiveness of such atmospheres in lowering the lethal temperatures relative to those in air. Effects on adult S. oryzae at 25 °C were explored in more detail in experiments using the following conditions: exposure to anoxic CAs for extended times; exposure to hypoxic CAs; and simulated hermetic storage. A simple scanning calorimetric method was developed for determining lethal temperatures and a combined thermo-gravimetric and differential thermal-analysis method was used to interpret the thermal events, due to loss of water, occurring at and above these temperatures.     

Distribution of the Medea factor M in populations of Tribolium castaneum (Herbst) in the United States

The distribution of the maternally acting, selfish gene Medea⁴ (M⁴) was determined in populations of the red flour beetle, Tribolium castaneum (Herbst), collected in the southern and midwestern United States. We found clear evidence for the existence of two major regional subpopulations, with a boundary that roughly corresponds to 33°N latitude. All 26 strains collected in 10 states north of this latitude were homozygous for the M⁴ allele, while only two of 29 strains collected in six states south of this latitude were homozygous for the allele. Of the remaining 27 southern strains, 21 lacked the M⁴ allele entirely, while six contained a mixture of M⁴ and non-M⁴ alleles. This is the first evidence of either the existence of biotypes or the presence of major barriers to gene flow in wild populations of this ubiquitous insect species.     

Progress towards a lure to attract three stored product weevils, Sitophilus zeamais Motschulsky, S. oryzae (L.) and S. granarius (L.) (Coleoptera: Curculionidae)

The behavioural responses of Sitophilus granarius, S. oryzae and S. zeamais to synthetic 4S,5R-sitophinone alone and in combination with volatiles from kibbled carob have been investigated with a view to the development of a single lure to attract all three species. Sitophilus zeamais and S. oryzae were shown to respond to 4S,5R-sitophinone at amounts as low as 0.025 ng. Sitophilus granarius gave a significant response to 50 ng 4S,5R-sitophinone. Volatiles from kibbled carob were also shown to attract all three species. This is the first time that attraction of S. zeamais and S. oryzae to carob volatiles has been demonstrated. Fresh lures containing 4S,5R-sitophinone and carob volatiles attracted significantly more insects in pitfall bioassays for all three species than when either component was used alone. However, after 6 weeks a significant increase in response compared to the control was found only for S. oryzae and S. zeamais. The effect of insect age on response to the pheromone and carob volatiles was also studied for all three species. The response to carob volatiles decreased with increasing insect age for all three species. A significant response to 4S,5R-sitophinone was found only for 1-2-day-old adults of S. oryzae and S. zeamais but for all ages tested of S. granarius. The effect of the combination of pheromone and carob volatiles also decreased with increasing insect age.     

The influence of fluctuating low-level magnetic fields on the fecundity and behaviour of Rhyzopertha dominica (F.)

Experiments were carried out to investigate the effects of fluctuating low-level magnetic fields on the demography and behaviour of Rhyzopertha dominica (F.). Adult insects were exposed to low-level magnetic fields, generated from both full- and half-wave rectified power supplies at 50 Hz and with magnetic-field strengths of 2-4 times that of the earth. Significant insect responses, in terms of changes to fecundity, development period and aspects of behaviour, were observed in many of the experiments and the nature of the response depended upon the power supply generating the magnetic field used.There appeared to be a dose effect because the reduction in progeny production diminished with distance from the centre of the field and the effects upon fecundity were cumulative. Numbers of progeny produced declined steadily over a 5-week period of exposure to the magnetic field, contrasting with the control which increased over the same period. The behavioural experiments showed that, despite a marked position effect within the experimental room, the magnetic field clearly had a deterrent effect upon the insects.The complex nature of the observed responses is discussed in relation to measured differences between the power supplies used in the experiments. The general issue of experimental methodology in such studies is also explored.     

Detection of Rhyzopertha dominica larvae in stored wheat using ELISA: The impact of myosin degradation following fumigation

Hard red winter wheat kernels were infested with eggs of Rhyzopertha dominica. After 20 d, when the larvae reached the fourth instar, they were killed by exposing the infested kernels to phosphine gas for 24 h. The infested kernels were then divided into four portions and treated as follows: one portion was immediately frozen at −80 °C to avoid myosin degradation; the other three portions were kept at 32 °C and 65% relative humidity, and then frozen at −80 °C after 14, 28, and 56 d post-fumigation, respectively. Each treatment was replicated five times. Myosin was measured using a commercial enzyme linked immunosorbent assay (ELISA) method that specifically detects this protein (Biotect^®, Austin, TX). Myosin degradation was most rapid in the first 2 weeks after the larvae were killed, decreasing from 1.672 to 0.695 ng/well during this period (a 58.4% reduction). There were no significant differences in myosin degradation between samples that were 14, 28, and 56 d post-fumigation. Grain is often fumigated to control insects. Frequently, this occurs many weeks before the grain is milled and may be repeated during the storage period. Therefore, estimates using the ELISA test may underestimate internal insect infestation because of myosin degradation. Insect fragment estimates for previously fumigated grain could be underestimated by as much as 58%.     

Response and genetic analysis of malathion-specific resistant Tribolium castaneum (Herbst) in relation to population density

Extensive use of malathion for pest control on stored cereals has resulted in worldwide resistance in red flour beetles, Tribolium castaneum. In this paper we investigate population density effects on the malathion-specific resistance in PRm, a strain from the Philippines, in an integrated resistance management framework. Two populations of malathion-specific resistant (PRm) and a malathion-susceptible strain of T. castaneum were bred at different densities: low (4 adults/g) and high (12 adults/g) density on wheat plus brewer's yeast in the laboratory. After eight generations, slopes of probit regression lines and LC₅₀ values were used to monitor the effect of insect rearing density on the progression of malathion-specific resistance. The LC₅₀ of the malathion-susceptible strain (Asm) did not change significantly during selection while LC₅₀s varied for both the high-density and low-density lines of PRm, the LC₅₀ of malathion ranged from 27.51 to 34.06 and from 21.14 to 29.39 μg malathion cm⁻² for high and low density, respectively.More than 33 generations were required to achieve a 10-fold increase of resistance for the low-density line compared to only 17 generations for the high-density line. Calculations from published formulae suggested that the malathion-specific resistance of both high- and low-density lines was under monofactorial control, with complete dominance.The data showed that environmental factors such as population density differences in insect rearing and development may influence the heritability of resistance. Furthermore, the variability in results published worldwide on resistance emphasises the need to standardize test conditions across laboratories.     

Density-dependent reproductive success in Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)

Two strains of Tribolium castaneum, one being susceptible to malathion (Asm) and the other malathion-specific resistant (PRm), were used to assess the effect of population density and its interaction with genetic background in reproductive success. A highly significant allometric relationship between female body weight and fecundity (R²=0.413) and another between female body weight and larval survivorship (R²=0.561) were found. Data showed that population density exerted an indirect effect on the reproductive success. The body weight of the Asm female was not affected by variations in population density but the body weight of the PRm female increased with reduced population density.The genetic background (strain) and/or malathion-specific resistance greatly influenced reproductive success. Concerning the percentage survivorship of offspring of the two strains, there was an opposite trend with increased population density: in PRm, survivorship was lower at high density (76.64±13.75; mean±SD) and higher at low density (88.39±7.61), whereas in Asm, survivorship was higher at high density (53.39±15.57) and lower at low density (43.99±20.08). The PRm female laid more eggs than the Asm. Reproductive success was significantly higher in PRm than in Asm. In addition, the genetic background (the strain) and/or the pleiotropic effect of malathion-specific resistance had a significant effect on the fecundity of PRm because the differences in fecundity and reproductive success between Asm and PRm went beyond the body weight of the female and might be explained by their genetic background and/or the pleiotropic effect of malathion-specific resistance.     

Efficacy of a volatile formulation of hydroprene (Pointsource™) to control Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae)

Three- and 4-week-old Tribolium castaneum (Herbst), the red flour beetle, and Tribolium confusum (du Val), the confused flour beetle, were exposed at five different temperature-relative humidity (r.h.) combinations to a volatile formulation of the insect growth regulator (IGR) hydroprene (called Pointsource™). Typical effects associated with IGR exposure, such as arrested larval growth, morphological deformities in adults, twisted and deformed wings, and incomplete adult emergence were produced in both species. Tribolium castaneum and T. confusum were susceptible to Pointsource™, but T. castaneum appeared to be the more susceptible species. More 3-week-old larvae of both species were arrested in that stage compared to the 4-week-old larvae. Nearly all of the 3- and 4-week-old T. castaneum larvae that were able to complete development to the adult stage quickly died after they emerged and were grossly morphologically deformed. In contrast, some emerged adult T. confusum remained alive after they emerged and were not deformed in any manner or had only twisted and incomplete wings. A greater percentage of larvae of both species were arrested in the larval stage and more adults died after they emerged in exposure studies conducted at 32°C, 75% r.h. as compared with 32°C, 30% r.h., but the reverse was true for exposures conducted at 27°C. Pointsource™ appears to have excellent potential for use in controlling Tribolium species within indoor facilities.     

Impact of high temperatures on efficacy of cyfluthrin and hydroprene applied to concrete to control Tribolium castaneum (Herbst)

In laboratory trials, concrete was treated with cyfluthrin wettable powder (WP) at 40 mg active ingredient [AI] cyfluthrin WP/m², then heated for 4, 8, or 16 h at either 45°C or 55°C, or treated but not heated (seven treatment combinations). Bioassays were conducted by exposing adult Tribolium castaneum (Herbst) for 0.5, 1, and 2 h. Survival of T. castaneum was generally greater on unheated concrete compared with the heating treatments, and survival appeared to decrease as heating time increased at both 45°C and 55°C. In a second laboratory trial, concrete was treated with hydroprene (Gentrol) at the label rate of 1.9×10⁻³ mg [AI]/cm², and bioassayed by exposing late-instar T. castaneum larvae on the treated surface. There were significant differences between untreated controls and the heat treatment regimes (P<0.05) with respect to the percentage of live emerged adults, the percentage of those adults with deformities, and the percentage of dead adults, but heating did not reduce efficacy of hydroprene. In a field trial, concrete was treated with cyfluthrin at 2 mg [AI] cyfluthrin WP/m², and placed in a flour mill undergoing an experimental heat treatment and in an unheated office. Treated concrete was bioassayed by continually exposing adult T. castaneum for 0.5-120 h. The effect of heating time on insect mortality was not significant (P?0.05). Except for T. castaneum exposed for 0.5 h, the percentage of beetle survival on unheated concrete was greater (P<0.05) than survival on concrete that had been heated in the mill, indicating a possible beneficial effect on cyfluthrin toxicity due to heating. Results of these studies show that short-term exposures to high temperatures may have no appreciable effect on efficacy of either cyfluthrin WP or hydroprene, and combination treatments of heat plus either of these insecticides may be effective alternatives to methyl bromide for disinfesting milling facilities.     

Insecticidal efficacy of diatomaceous earth against Sitophilus oryzae (L.) (Coleoptera: Curculionidae) and Tribolium confusum du Val (Coleoptera: Tenebrionidae) on stored wheat: influence of dose rate, temperature and exposure interval

Laboratory experiments were conducted in order to assess the insecticidal effect of a diatomaceous earth formulation (Silicosec^®, Biofa GmbH, Germany) against Sitophilus oryzae and Tribolium confusum on stored wheat. Adults of the two species were exposed on wheat treated with diatomaceous earth at four dose rates: 0.25, 0.5, 1 and 1.5 g/kg of wheat, respectively. For each dose rate, the treated wheat was placed at 22°C, 25°C, 27°C, 30°C and 32°C. Dead adults were counted after 24 and 48 h, 7 and 14 d of exposure. After the 14-d interval, the live adults were removed and placed for 7 d in untreated wheat (in the case of S. oryzae) or untreated flour (in the case of T. confusum), and the production of F₁ was recorded. For both species, dose rate, temperature and exposure interval significantly affected mortality (P<0.001). Mortality was higher at longer exposure intervals. The efficacy of SilicoSec against S. oryzae increased with temperature, but for T. confusum mortality was lower at 32°C, compared to 30°C, for 24 and 48 h exposure intervals. Tribolium confusum proved less susceptible to SilicoSec than S. oryzae. In general, the rates of 1 and 1.5 g/kg of wheat provided a satisfactory level of protection against the two species examined. For S. oryzae, F₁ emerged only at 22°C, in wheat treated with 0.25 or 0.5 g/kg. However, for T. confusum, F₁ were recorded at 22°C for 0.5 g/kg and at 22°C, 25°C, 27°C and 30°C for 0.25 g/kg.     

Survival of Sitophilus oryzae (L.) on wheat treated with diatomaceous earth: impact of biological and environmental parameters on product efficacy

A series of experiments was conducted to evaluate the effects of temperature, relative humidity (r.h.), population density, concentration, exposure interval, and residual aging on susceptibility of Sitophilus oryzae (L.), the rice weevil, to diatomaceous earth (DE). In the first experiment, hard red winter wheat was treated with 300 ppm of the Protect-It™ formulation of DE, and 10, 20, or 30 1-2 week-old mixed-sex adult weevils were exposed on 35 g of wheat for 1 week at combinations of 22°C, 27°C, or 32°C; 40%, 57%, or 75% r.h. No weevils survived when exposed at 40% or 57% r.h., but at 75% r.h. survival was related to both population density and temperature. A higher percentage of adults survived when 30 were exposed compared to 10 and 20, and within each density, survival decreased with increasing temperature. No F₁s were produced at any r.h. on wheat held at 22°C. At 27°C and 32°C, the maximum number of F₁s was produced on wheat held at 75% r.h. In the second experiment, wheat was treated with 25%, 50%, 75%, or 100% of the label rate of 300 ppm, and 10 mixed-sex adult S. oryzae were exposed on 35 g of wheat for either 1, 2, or 3 weeks at 27°C, 57% and 75% r.h. Survival decreased with increasing exposure interval and concentration, but within exposure interval and concentration, survival was usually greater at 75% versus 57% r.h. In the final experiment, wheat was treated with 300 ppm, held at 22°C and 27°C, 57% r.h., and bioassayed at monthly intervals for 3 months by exposing 20 adult mixed-sex S. oryzae on 35 g of wheat for 1 or 2 weeks. At each month, survival of S. oryzae was greater when exposed at 22°C compared to 27°C and when exposed for 1 week compared with 2 weeks. Survival gradually increased with each monthly bioassay, except for those conducted at 3 months. Results of these studies show that S. oryzae is susceptible to DE, but survival of exposed insects will depend in part on the temperature and r.h. humidity (or grain moisture content) at which they are exposed. Survival is directly related to temperature, and as r.h. increases either higher concentrations or longer exposure intervals will be necessary to maintain a certain level of mortality. There may also be a loss of efficacy with residual aging.     

Impact of physical and biological factors on susceptibility of Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) to new formulations of hydroprene

Three separate experiments were conducted to: (1) evaluate two new commercial formulations (202-080 and 202-084) of the insect growth regulator hydroprene, (2) determine residual efficacy of hydroprene-treated concrete stored at different environmental conditions, and (3) assess the impact of accumulated flour on residual efficacy. In the first test, late instars of Tribolium castaneum (Herbst), the red flour beetle, and Tribolium confusum (du Val), the confused flour beetle, were exposed on concrete treated with hydroprene. At 40% relative humidity (r.h.), there was no difference between species regarding the percentage of individuals that stopped development in the larval stage (arrested larvae), but at 75% r.h. there were more arrested T. castaneum than T. confusum in all treatments except the low rate of formulation 202-084. No adult T. castaneum lived after emergence (live adults) at either relative humidity, but the percentage of live adult T. confusum ranged from 1.0% to 41.0%, depending on treatment. In the second test, late instars of T. confusum were exposed at 6 and 12 weeks post-treatment on concrete treated with the two experimental formulations and stored under different environmental conditions. At 6 weeks there was no difference between formulations. At 12 weeks, fewer live adults and more dead emerged adults with gross morphological deformities were found on concrete treated with formulation 202-084 and stored at 32°C, 75% r.h. compared to other treatment combinations. In the final experiment, wheat flour was added to treated concrete for 5 weeks before the bioassays were conducted with late-instar T. confusum. There were few live adults produced in the initial bioassays, and dead adults with gross morphological deformities ranged from 83.1% to 97.6%. However, in bioassays conducted with late-instar larvae at 6 weeks, most adults eventually emerged with few deformities. The presence of the flour apparently compromised residual control and the hydroprene was no longer effective. In summary, the new hydroprene formulations were equivalent to the registered product Gentrol^®. Tribolium confusum was less susceptible than T. castaneum, and residual control of T. confusum on a clean surface without flour lasted about 6-12 weeks.     

Initial and delayed mortality of late-instar larvae, pupae, and adults of Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) exposed at variable temperatures and time intervals

Late-instar larvae, pupae, and adults of Tribolium castaneum (Herbst), the red flour beetle, or Tribolium confusum (DuVal), the confused flour beetle, were exposed for variable durations at 36-54 °C. Beetles were placed in laboratory ovens set at a baseline of 27 °C, the temperature was increased by 0.1 °C per minute until the target temperature was achieved, and beetles were then held for specified exposure durations. There was no mortality after initial exposure or after a 1-week holding period of any life stage of T. castaneum or T. confusum exposed for 32 h to 36, 39, or 42 °C. At 45 °C, there was no initial mortality of either species exposed for different time intervals except for those exposed for 28 h. However, there was a significant increase in mortality after the 1-week holding period of those beetles exposed initially for at least 16 h to 45 °C. There was a sharp increase in mortality after the initial exposures of 4 h at 48 °C; mortality of T. confusum larvae was 90.0±5.7% but was only 10.0±10.0% for larvae of T. castaneum, and no pupae of either species were dead. All life stages of both species were killed after the initial exposure of 12 h, and 1-week mortality of beetles exposed for 4 and 8 h was generally greater than initial mortality. At 51 and 54 °C, 2- and 1-h exposures, respectively, killed all life stages of each species. Mortality in conditions of gradual temperature increase was less than previous studies with sudden temperature increases.     

Grain loss caused by Tribolium castaneum, Sitophilus oryzae and Acanthoscelides obtectus in stored durum wheat and beans treated with Beauveria bassiana

The effect of the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin (Hyphomycete) on the losses caused to durum wheat and beans by storage insects was investigated. Grains were infested with Tribolium castaneum (Herbst), Sitophilus oryzae (L.) and Acanthoscelides obtectus (Say). Beauveria bassiana was produced in inoculated autoclaved rice. The spore formulation (ground rice and B. bassiana) was applied to grain (wheat or bean) and shaken to evenly cover the grain. Adults of T. castaneum or S. oryzae were added to wheat and adults of A. obtectus to bean. Five replicates were set up for each treatment and controls (milled rice but no conidia). The insecticidal effect of B. bassiana was tested by measuring the fresh weight and weight loss of grains after four months of storage. Wheat grains infested with S. oryzae without the conidia was significantly more damaged by weevils than grains treated with B. bassiana. The mean fresh weight of grains with the conidia was significantly greater (18.4%) than the corresponding mean without the fungus when S. oryzae were present. Percentage weight loss decreased by 81.5% and was significantly smaller than the loss from the untreated grain. Significant differences were not found in the fresh weight of seed infested with T. castaneum or A. obtectus in treated or untreated grain nor in the percentage weight loss of grains infested with these insects, with and without B. bassiana.