Evaluation of three new insecticide formulations based on inert dusts and botanicals against four stored-grain beetles

Three new insecticide formulations were developed that combine diatomaceous earth (Celatom MN 23) with compounds with low mammalian toxicity. Sitophilus oryzae, Sitophilus granarius, Rhyzopertha dominica and Tribolium castaneum were held on wheat (13.5% moisture content) treated with different doses of the formulations at 28 ± 1 °C and 60 ± 5% RH for 2–7 days, and then held for progeny emergence. The three formulations were made up of: dill essential oil, bait, silica gel and DE (Dill-DE); pyrethrin, PBO, dill essential oil, silica gel and DE (Py-Dill-DE); and disodium octaborate tetrahydrate (DOT), pyrethrin, PBO, dill essential oil, silica gel and DE (DOT-PY-Dill-DE). They were applied either as powders or as slurries. Applying the formulations as powders over a range of concentrations showed that pure DE is the least effective insecticide, with 50% mortality ranging from 430 to 1000 ppm depending on the species after 2–3 days. Dill-DE formulation was the next least effective formulation. The Py-Dill-DE and DOT-Py-Dill-DE formulations had similar activity and being the most effective formulations with 50% mortality from 40 to 260 ppm depending on the species. Bioassays were also run at single doses. Dill-DE was applied at 300 ppm, Py-Dill-DE at 150 ppm and DOT-PY-Dill-DE at 200 ppm. After 3 days, Py-Dill-DE and DOT-PY-Dill-DE formulations there was 100% mortality for S. oryzae, S. granarius and R. dominica. After 7 days the mortality of T. castaneum was from 96 to 100%. Both formulations reduced the progeny almost by 100%. There was survival for all insects with Dill-DE formulation. The concentrations of 150 ppm of Py-Dill-DE, 200 ppm of DOT-PY-Dill-DE and 300 ppm of Dill-DE reduced bulk densities by 0.7, 0.8 and 5.7 kg/hL, respectively.     

Efficacy of diatomaceous earths and their low-dose combinations with spinosad or deltamethrin against three beetle pests of stored-maize

A key element in postharvest IPM is the reduction of chemical residues in food through the use of reduced dosages of less toxic grain protectants. Two laboratory experiments were conducted: Experiment I determined the efficacies of straight diatomaceous earths (DEs) – “Chemutsi” (African raw DE), MN51 (new formulation) and Protect-It^® (enhanced DE), and two new food grade DE-based formulations (A2 and A3) against adult Prostephanus truncatus (Horn), Sitophilus zeamais (Motschulsky) and Tribolium castaneum (Herbst) admixed with shelled maize. In Experiment II, Chemutsi and Protect-It^® were further tested in varying combinations with low-dose deltamethrin and spinosad. At 21 days post-exposure, MN51 800 ppm and 1000 ppm, Chemutsi 1000 ppm, Protect-It^® 600 ppm and food grade A3 150 ppm caused S. zeamais mortalities that were not significantly different from the positive control (Protect-It^® 1000 ppm). However, after the same exposure period, all the straight DEs (applied at ≤ 1000 ppm) and the DE-based food grade formulations were not effective on P. truncatus and T. castaneum. In low dose combinations, 7 day mortalities showed high S. zeamais susceptibility to both DE-spinosad and DE-deltamethrin while P. truncatus was more susceptible only to DE-spinosad and T. castaneum to Protect-It^®-deltamethrin only. At 21 days, all DE-spinosad and DE-deltamethrin treatments were effective and not significantly different from the commercial grain protectant (fenitrothion 1.0% w/w (10000 ppm) + deltamethrin 0.13% w/w (130 ppm)) on all test species. DE-spinosad and DE-deltamethrin combinations significantly suppressed (P < 0.001) F₁ progeny for the three test species whereas straight DEs and DE-based food grade formulations did not. Our results showed that at half the label rates or lower, DE-spinosad and DE-deltamethrin combinations were effective alternative grain protectants that are safer and possibly cheaper. We also give the first report on the effectiveness of Chemutsi in combination with spinosad or deltamethrin on maize grain.     

Responses of phosphine susceptible and resistant strains of five stored-product insect species to chlorine dioxide

Adults of phosphine susceptible laboratory strains and phosphine resistant field strains of five stored-product insect species were exposed in vials with 0 or 10 g of wheat for different time periods to 0.54 g/m³ (200 ppm) of chlorine dioxide gas. After exposure, adult mortality was determined 5 d later at 28 °C and 65% r.h. The 5-d mortality was 100% in laboratory and field strains of the red flour beetle, Tribolium castaneum (Herbst); sawtoothed grain beetle, Oryzaephilus surinamensis (L.); lesser grain borer, Rhyzopertha dominica (F.); maize weevil, Sitophilus zeamais Motschulsky; and rice weevil, Sitophilus oryzae (L.) that were exposed in vials with 10 g of wheat to chlorine dioxide for 26, 16, 24–34, 18–24, and 15–18 h, respectively. Corresponding exposure durations for these species and strains in vials without wheat were 15, 3, 18–20, 7–15, and 5–7 h, respectively. Dosages of chlorine dioxide producing 99% mortality (LD₉₉) of T. castaneum, O. surinamensis, R. dominica, S. zeamais, and S. oryzae strains in vials with wheat ranged from 14.79−22.57, 8.20–8.41, 15.79–21.60, 10.66–14.53, and 7.67–12.20 g-h/m³, respectively. In vials without wheat, corresponding LD₉₉ values for T. castaneum, R. dominica, and S. zeamais strains were 6.51–8.66, 11.46–23.17, and 5.79–10.26 g-h/m³, respectively. LD₉₉ values for O. surinamensis and S. oryzae could not be computed, because of 100% mortality after a 3–5 h exposure to chlorine dioxide. No adult progeny production of T. castaneum and O. surinamensis was observed after 8 weeks in control and chlorine dioxide-exposed samples. Adult progeny production of Sitophilus spp. was found only in the control samples. The dosage for 99% adult progeny reduction relative to control for R. dominica strains ranged from 10.07 to 18.11 g-h/m³. Chlorine dioxide gas is effective in killing phosphine susceptible and resistant strains of five stored-product insect species and suppressing adult progeny production of three out of the five species.     

Efficacy and adherence ratio of diatomaceous earth and spinosad in three wheat varieties against three stored-product insect pests

Laboratory tests were carried out in order to evaluate the efficacy of three diatomaceous earth (DE) formulations; Protector, SilicoSec, and Insecto, and one spinosad dust formulation in three commercially available wheat (Triticum durum) varieties, Athos, Pontos, Sifnos, originating from Greek cultures. The efficacy of the above formulations was assessed against adults of Rhyzopertha dominica, Sitophilus oryzae and Tribolium confusum. The DE formulations were applied at three dose levels; 250, 500 and 1000 ppm, while spinosad was applied at 100, 500 and 1000 ppm of the formulation, corresponding to 0.125, 0.625 and 1.25 ppm active ingredient (a.i.), respectively. The adherence ratio of the DE and spinosad formulations to the kernels of the wheat varieties was assessed. Bioassays were carried out at 30°C and 60% r.h. Mortality was recorded after 7 and 14 d in the case of R. dominica and S. oryzae, and after 7 d, 14 d and 21 d of exposure in the case of T. confusum, on the treated varieties. Progeny production of the individuals exposed on the treated varieties was also assessed. Even the lowest dose of spinosad was highly effective (>90%) against R. dominica and S. oryzae. In the case of T. confusum a combination of longer exposures with higher doses was required for each formulation to be effective. Generally, the performance of all formulations was greater in Athos or Sifnos compared to the Pontos. Progeny production of S. oryzae and R. dominica was significantly higher in untreated Pontos than in the other varieties. Progeny production was always greater in untreated than treated substrates. Although adherence ratios for the tested DE and spinosad formulations were generally high (>90%), significant variations in adherence levels among the wheat varieties were recorded.     

Distance and height of attraction by walking and flying beetles to traps with simultaneous use of the aggregation pheromones from Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae)

Adults of Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.) biosynthesize aggregation pheromones 4,8-dimethyldecanal and dominicalure-1/dominicalure-2, respectively. These pheromones are commonly used independently, and their simultaneous use has not been adequately studied. Furthermore, information on trapping flying R. dominica in pheromone traps is minimum. Therefore, the objectives were to evaluate distance of attraction of T. castaneum and R. dominica adults to traps having both pheromones, and height of R. dominica adult attraction to traps with its pheromone lure alone. In first experiment, both pheromones were deployed simultaneously inside a commercial pitfall trap. One-month-old 20 T. castaneum and R. dominica adults were released every 30 cm from the pheromone trap. The adults trapped were recorded at 4 and 24 h following their release. Adults of both species released were captured in higher percentages at 24 h than 4 h. At 30 cm distance, these values were 45.5% for T. castaneum and 10–12% for R. dominica for 24 h whereas they were 40.5% for T. castaneum and 5–7.5% for R. dominica following 4 h exposure. The maximum trap capture was at 30 cm for T. castaneum and 30–60 cm for R. dominica. In second experiment, a trap with two rubber septa containing dominicalure-1 and dominicalure-2 was placed at different heights inside cage, and R. dominica adults were released at the bottom. In each experiment, four replicates were tested. After 24 h, flying R. dominica adults were captured in progressively lower percentages as trap height increased up to 40 cm above the bottom of cage. Trap heights above 10 cm exhibited decreased trap capture of R. dominica compared to those at 3 cm. We conclude that simultaneous use of both aggregation pheromones better facilitates trapping of walking T. castaneum and R. dominica. Traps above 10 cm show decreased captures of flying R. dominica.     

Testing the consistency of spatio-temporal patterns of flight activity in the stored grain beetles Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.)

Many studies have been published on the flight activity of stored grain beetles in temperate regions, but few have focussed on tropical and sub-tropical regions. The spatio-temporal patterns of flight activity of Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.) were quantified in a grain growing district on the Tropic of Capricorn in central Queensland, Australia. Nine traps baited with aggregation pheromone lures were monitored at 2-wk intervals (fortnightly) for 1 year along a 28.4 km linear transect that included sites at bulk grain depots and sites away from stored grain. Beetles of both species were trapped every fortnight during the study. The spatio-temporal patterns of flight activity differed greatly across the two species, as predicted from studies elsewhere. Rhyzopertha dominica was widespread across the landscape, as the mean trap catch of this species was equal in depot and non-depot traps. In contrast, T. castaneum was more frequently trapped in depot traps than non-depot traps during the colder months, but was much more widespread across the landscape during the summer months. Tribolium castaneum also showed a clear mid-summer peak in flight activity, whereas R. dominica flight activity was highly variable throughout the study. In general, our results reveal patterns that are consistent with those found for T. castaneum and R. dominica in southern Queensland. The contrasting spatio-temporal patterns of flight activity of R. dominica and T. castaneum show that species-specific approaches may be needed to manage these pests and that the spatio-temporal dynamics of resistance genes may differ across these species.     

Effects of methoprene and synergized pyrethrin aerosol applications on Tribolium castaneum (Herbst) populations

The occurrence of horizontal transfer of the insect growth regulator (IGR) methoprene on confined populations of Tribolium castaneum (Herbst), either with or without hidden refugia, was determined through a series of experiments. Multiple applications were made with the IGR alone or combined with synergized pyrethrin, and compared to untreated controls which received no insecticide applications or were treated with the carrier Isopar M that was a component of the pyrethrin formulation. The total number of living beetles from test colonies inoculated with adults that were treated with Isopar M or with no adults (control) was significantly greater than those colonies that were inoculated with adults treated with either synergized pyrethrin or methoprene (P < 0.05). There was no difference in the number of living individual in the larval, pupal, and adult stages and the instantaneous rate of increase (r_i) in established populations treated with methoprene and containing a hidden refugia compared to those which received the pyrethrin applications (P ≥ 0.05). Sanitation levels of two different flour quantities nested inside the treatment groups also had no effects (P ≥ 0.05), suggesting that populations in hidden refugia can persist even with multiple applications of methoprene and synergized pyrethrin. Populations with an accessible hidden refugia that were exposed to synergized pyrethrin and methoprene had a lower number of living adult and a lower r_i value than populations that were exposed to synergized pyrethrin alone (P < 0.001). Additionally, populations which received one, two, or three aerosol applications had similar numbers of living adults and r_i but were significantly different from populations which received four aerosol applications. Results suggest multiple applications of methoprene and synergized pyrethrin could be more effective than synergized pyrethrin alone for control of T. castaneum.     

Laboratory assessment of insecticidal effectiveness of natural zeolite and diatomaceous earth formulations against three stored-product beetle pests

The insecticidal effectiveness of two natural zeolite formulations (Minazel plus and Minazel), applied to wheat at selected rates of 0.25, 0.50 and 0.75 g/kg, and a diatomaceous earth formulation (DE) (Protect-It™), applied at the recommended rates of 0.15 g/kg for Sitophilus oryzae, 0.20 g/kg for Rhyzopertha dominica and 0.30 g/kg for Tribolium castaneum, were tested under laboratory conditions (24 ± 1 °C temperature and 45 ± 5% relative humidity). The highest adult mortality was observed after the longest exposure period of 21 days and 7 days of recovery, when all three zeolite dosage rates and the recommended DE dosage caused 97-100% mortality of S. oryzae and 94-100% of T. castaneum. On the other hand, 100% mortality was not achieved in any test variant involving R. dominica; the highest (about 92%) was detected for DE, while 52% and 79% mortality was achieved with the zeolites at the highest rate of 0.75 g/kg. Progeny reduction by >90% was achieved after 21 days of contact of all three beetle pests with DE-treated wheat, while the same level of reduction was achieved for S. oryzae and T. castaenum only after contact with the highest rate of the zeolite product, Minazel. Thus the two zeolite formulations are comparable to diatomaceous earth in controlling adult S. oryzae, R. dominica and T. castaneum, but only the Minazel formulation could effectively protect wheat from attack by S. oryzae or T. castaneum, and only with a higher rate of application than for the DE formulation.     

Solvent free microwave extraction followed by encapsulation of O. basilicum L. essential oil for insecticide purpose

Currently essential oils are increasingly used as natural insecticides because of the adverse effects on human health and environmental pollution of synthetic insecticides. The aim of this study is to perform the encapsulation of Ocimum basilicum L. essential oils by freeze-drying technique using binary mixtures maltodextrins and acacia gum to investigate their insecticidal activity against three insect pests of stored-products Rhyzopertha dominica, Sitophilus oryzae and Tribolium castaneum by direct contact and ingestion toxicity effects. Essential oils were obtained by solvent-free microwave extraction (SFME) and conventional hydro-distillation (HD) methods.Chemical composition of essential oils analysed by GC-MS indicated that the major components were linalool (43.5% for SFME and 48.4% for HD) followed by methyl chavicol (13.3% for SFME and 14.3% for HD) and 1,8-cineole (6.8% for SFME and 7.3% for HD). The toxic effects results, confirm that O. basilicum essential oils capsules were highly effective against R. dominica than S. oryzae, except for T. castaneum which was insensitive. The toxicity against adults of R. dominica varied between (LD₅₀ = 0.20 g/kg for SFME-EEO, LD₅₀ = 0.25 g/kg for HD-EEO) and (LD₅₀ = 0.32 g/kg for SFME-EEO, LD₅₀ = 0.37 g/kg for HD-EEO) by direct contact and ingestion effect, respectively. These results lead us to consider such essential oils capsules as providing a richer source of natural insecticide agents for food preservation.     

Toxicity of coumaran to stored products beetles

The fumigant activity of coumaran was tested against mixed-age cultures of the rice weevil (Sitophilus oryzae L.), the lesser grain borer (Rhyzopertha dominica F.), the rust-red flour beetle (Tribolium castaneum Herbst), saw-toothed grain beetle (Oryzaephilus surinamensis L.) and flat grain beetle (Cryptolestes ferrugineus Stephens). T. castaneum was tolerant to coumaran, for total population extinction was achieved in 72 h exposure at 50 μg/L dose only. In contrast all developmental stages of O. surinamensis, C. ferrugineus and R. dominica were quite susceptible 100% mortality was attained at < 50 μg/L doses in 24 or 72 h exposure. Further results showed that T. castaneum, generally tolerant pest to fumigants and R. dominica, a predominant stored grain pest in India, could be controlled by coumaran. The findings indicate that, coumaran is good source for alternate to methylbromide for effective control of stored grain insect pests.     

Toxicity and growth inhibitory activities of methanol extract and the β-carboline alkaloids of Peganum harmala L. against two coleopteran stored-grain pests

Methanol extract and the β-carboline alkaloids were extracted from the seeds of Peganum harmala (Zygophyllaceae). Their toxicity, growth inhibitory and effects on the progeny production of Tribolium castaneum and Rhyzopertha dominica was studied. To assess any additive effects among the extracted β-carbolines, they were tested as binary mixtures (1:1) or as a crude alkaloid fraction. All extracts exhibited a considerable adulticidal effect with increasing activities in response to increased exposure period. Using the contact toxicity bioassay, the crude β-carboline fraction was the most effective (LC_50’s were 20.1 and 36.7) μg/cm², 48 h post-treatment against R. dominica and T. castaneum, respectively. LC_50’s of (harmaline + harmine), (harmaline + harmane), and methanol extract were (31.2, 39.4), (33.7, 47.2), and (39.8, 65.2) μg/cm², 24 h post-treatment against R. dominica and T. castaneum, respectively. At 48 h post-treatment, LC₅₀ of (harmaline + harmine) reached 22.4 μg/cm² against R. dominica. When mixed with the insect’s diets, toxicity of all extracts were increased with the crude alkaloidal fraction the most toxic (LC_50’s were 7.8 and 14.7) mg/kg grains, 48 h post exposure against R. dominica and T. castaneum, respectively. When the 2nd instar larvae were fed sub-lethal doses-treated grains, development and F1 progeny of both insects were significantly affected (P ≤ 0.001). At 3.5 mg/kg grains of the crude alkaloidal extract, percentages of malformed larvae and pupae of T. castaneum were 19.7 and 33.4%, respectively. In this case, a total life span of 81.3 days was recorded for the treated individuals compared to 44.2 for the control. A reduction in the adult progeny of 56.9, 44.0 and 43.6% was obtained with 3.5 mg/kg of the crude alkaloids, (harmaline + harmine) and methanol extract, respectively. Meanwhile, the reduction in adult progeny of R. dominica reached 79.2% with the same concentration of the crude alkaloid extract.     

Efficacy of methoprene for multi-year protection of stored wheat,brown rice,rough rice and corn

Hard red winter wheat, brown rice, rough rice, and corn were treated with the insect growth regulator (IGR) methoprene at rates of 1.25 and 2.5 ppm, held for 24 months at ambient conditions in buckets on the floor of a grain bin, and sampled every two months. Bioassays were done by exposing 10 mixed-sex adults of Rhyzopertha dominica (F.), the lesser grain borer, and Tribolium castaneum (Herbst), the red flour beetle, on wheat, R. dominica and Sitotroga cerealella (Oliver), the Angoumois grain moth, on brown rice and rough rice, and T. castaneum and S. cerealella on corn. Sample size for all commodities was about 80 g, and these samples were held for 3 months at 27°C-60% r.h. Both rates of the IGR completely suppressed adult progeny development of R. dominica with little resulting feeding damage, sample weight loss, or insect damaged kernels (IDK). Some adult progeny production of S. cerealella and resulting IDK occurred at both rates on rough rice, brown rice, and corn, but was far less than in untreated controls. There was little adult progeny production but some feeding damage caused by larval T. castaneum in the treated wheat and corn but again far less than in untreated control. Allowing continual exposure of parental adults on grains treated with an IGR, rather than exposing those parental adults for a short time period, may give more accurate evaluations of residual efficacy. Results show that methoprene used as a grain protectant will give residual control of stored product beetles for 24 months, but complete control of S. cerealella may require inclusion of a contact insecticide.     

Effectiveness of diatomaceous earth for control of Sitophilus zeamais (Coleoptera: Curculionidae), Tribolium castaneum and Palorus subdepressus (Coleoptera: Tenebrionidae)

The effectiveness of diatomaceous earth (DE) or diatomite has been assessed against three major beetle pests of stored maize: Sitophilus zeamais (the maize weevil), Tribolium castaneum (the red flour beetle) and Palorus subdepressus (the depressed flour beetle). Maize has been treated with four doses of DE (1.5, 3, 4.5 and 6 g/kg) and four doses of Actellic Super™ Dust a chemical insecticide used as a reference (0.25, 0.5, 0.75 and 1 g/kg). The effective mortality was measured for each treatment after 1, 2, 4, 7 and 14 d of exposure. The corrected mortality rates were calculated by considering the mortality in the control group. Diatomaceous earth was as effective as Actellic Super™ Dust but required higher doses. Diatomaceous earth acted faster on S. zeamais and P. subdepressus compared to T. castaneum. The highest dose tested for DE caused the same mortality in T. castaneum as the recommended dose of Actellic Super™ Dust. Diatomaceous earth is a good alternative for the control of these three devastating insect pests of maize stocks and can readily be incorporated into integrated stored products pest management programs.     

Performance of diatomaceous earth and imidacloprid as wheat,rice and maize protectants against four stored-grain insect pests

Laboratory experiments were conducted in Pakistan to investigate the effectiveness of the diatomaceous earth (DE) formulation, Protect-It, at 150 ppm and imidacloprid, at 1.25, 2.5 and 5.0 ppm, alone or in combination, against the rusty grain beetle, Cryptolestes ferrugineus Stephens (Coleoptera: Laemophloeidae), the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and the psocid, Liposcelis paeta Pearman (Psocoptera: Liposcelididae). The bioassays were conducted at 25 °C, 65% relative humidity (r.h.) and continuous darkness by using adults of each insect pest that were exposed on treated or untreated wheat, rice and maize. Adult mortality counts were performed 1, 2, 4 and 7 days post-exposure. Progeny production was also evaluated 62 and 30 days after the last mortality count for beetles and psocid respectively. Results showed that mortality was increased with the increase of dose and exposure interval. Imidacloprid was more effective at the highest dose than the DE alone. The combination of DE plus imidacloprid resulted to higher mortalities than DE or imidacloprid alone at almost all exposure intervals. There was higher mortality on wheat than on rice or maize. In general, T. castaneum was the most tolerant insect species to any treatment while L. paeta was the least tolerant. The highest progeny production was observed for T. castaneum (i.e., 33.1 individuals per vial) while the lowest for L. paeta (i.e., 11.1 individuals per vial) on maize treated with Protect-It alone. The findings of the current study indicate that the combined use of DE and imidacloprid, as wheat, rice and maize protectants, may provide adequate level of management to several important stored-product insects.     

Behavior of female Rhyzopertha dominica (Coleoptera: Bostrichidae) in a mono-layer of wheat treated with diatomaceous earth

Adult female lesser grain borers (Rhyzopertha dominica [F.]) were observed in a mono-layer of wheat sandwiched between two layers of glass to determine if movement patterns and survival rates differed in wheat that was admixed with diatomaceous earth (DE) compared with untreated wheat. Observations were also made to determine if responses to DE differed depending on the commercial formulation of DE tested at the labeled rates. Mortality was higher in the DE treatments than in the untreated controls, and also varied according to the DE formulation. In wheat treated with 1000 ppm Dryacide^®, 400 ppm Protect-It^®, and 500 ppm Insecto™ (labeled rate for the individual products), mean percentage mortality was 100±0, 71.4±10.1, and 57.1±11.1, respectively. Although total distance traveled and the number of turns taken by the beetles was lower in the Dryacide treatment compared to the untreated controls, the movement patterns were not significantly different among the three DE treatments. Observed differences in mortality are likely related to DE products or amount applied rather than to differences in DE exposure resulting from movement behavior.     

Toxicity and repellent action of Coffea arabica against Tribolium castaneum (Herbst) adults under laboratory conditions

Coffea arabica parchment extracts and caffeine isolated from the plant were evaluated randomly against 1-week-old adults of red flour beetle, Tribolium castaneum for fumigant toxicity and repellent action under laboratory conditions. The effects on detoxification enzymes and neuroenzyme was also determined. Among the various extracts prepared sequentially using hexane, dichloromethane, ethyl acetate and methanol as solvents, the dichloromethane extract did repel insects on contact (EC₅₀ = 4380.65 ppm). The repellence was more prominent when an olfactory response was measured (EC₅₀ = 2571 ppm). The active ingredient isolated from dichloromethane extract was identified as caffeine which showed very strong repellency as compared to the extract. In terms of toxicity of this extract, a significant mortality was recorded in fumigation assay (LC₅₀ = 5555 and 791 ppm, 24 and 48 h post-treatment, respectively). However, caffeine did not induce similar toxicity as the dichloromethane extract. The studies on the impact on detoxification enzymes of T. castaneum showed that dichloromethane extract inhibited carboxyl esterase activity, which possibly led to high toxicity. However, caffeine inhibited glutathione-s-transferase and induced carboxylesterase enzymes. It was, therefore, obvious that C. arabica parchment crude extracts have dual effects against T. castaneum adults, i,e, fumigant toxicity and repellent effects. However, the active compounds responsible for the two activities are surely different as caffeine could only induce repellent action against the beetles and the toxic compound needs to be identified, which is presently being investigated.     

Field trials on the efficacy of Beauveria bassiana,diatomaceous earth and Imidacloprid for the protection of wheat grains from four major stored grain insect pests

We assessed the insecticidal efficacy of Beauveria bassiana, diatomaceous earth (DE) and the neonicotinoid Imidacloprid against four major insect species of stored grain pests during field trials conducted on small farms located in four districts of Punjab, Pakistan. In each district, a 40 kg lot of wheat grain was admixed with B. bassiana (3 × 10¹⁰ conidia kg⁻¹), DE (150 ppm) and Imidacloprid (5.0 ppm) alone and in different combinations. Each lot was divided subsequently into four equal parts (10 kg each), packed in polypropylene bags, artificially infested, labeled and stored at the farms under natural environmental conditions up to 6 months. Sampling was carried out every 30 d to record the total number of dead and alive adults, as well as the percent of grain damage in treated and untreated (control) grains. Results revealed a significant difference among the treatments, test insect species and the storage period. For each district, the combined treatments provided better control of all tested insect species compared with each treatment alone. The least number of surviving adults and minimum percent grain damage was observed for Imidacloprid and DE combination, but was outperformed by the DE treatment with B. bassiana for long-term protection. For all grain protectants, we obtained the same order of susceptibility level among the test species (i.e. Liposcelis paeta > Cryptolestes ferrugineus > Rhyzopertha dominica > Tribolium castaneum). The results of the present study suggested that DE, insect pathogenic fungi and Imidacloprid can be effectively used for the protection of wheat stored at small-scale farmer's fields.     

First report of strong phosphine resistance in stored grain insects in a far northern tropical region of Australia,combining conventional and genetic diagnostics

Although phosphine resistance monitoring and management programs are well advanced in Australia, the tropical northern part of the continent has received little attention. To address this, grain storages were sampled in the Townsville region in the Burdekin river catchment area of northern Queensland, and insect populations were subjected to resistance testing. Sampling of storages including silos, food processing facilities and feedlots was carried out during July to August 2019 and February to March 2020. Of the several species collected, populations of two major pest species, Rhyzopertha dominica and Tribolium castaneum were subjected to phenotypic resistance and molecular screening using discriminating doses and gene specific DNA marker, respectively. Utilising phenotypic assay, of the 17 populations of R. dominica, 12 were diagnosed as weakly resistant and five as strongly resistant; whereas of the 34 populations of T. castaneum, two were found to be susceptible to phosphine, 15 were determined as weakly resistant and 17 as strongly resistant. The molecular diagnostics, however, determined rph2 alleles for strong resistance in some populations each of R. dominica and T. castaneum that were diagnosed as weakly resistant in the phenotypic assay. The most prevalent rph2 alleles were found to be P49S and K142E in R. dominica populations and P45S and G131S in T. castaneum populations. Our results highlight the need of more comprehensive study towards determining the level of pests and resistance risks in this region.     

Isofuranodiene-based nanoemulsion: larvicidal and adulticidal activity against tenebrionid beetles attacking stored wheat

The resistance of insect populations to the existing insecticides grows fast due to their continuous use. Thus, the need for new and effective formulations is increasing. Isolated compounds of botanical origin may represent a promising solution for green pest management strategies. In this study, isofuranodiene derived from Smyrnium olusatrum L. (Apiales: Apiaceae) essential oil (EO), was nanoemulsified (3% w/w) to treat wheat kernels with two concentrations (500 and 1000 ppm) against adults and larvae of three tenebrionid beetles attacking stored products, the red flour beetle, Tribolium castaneum (Herbst), the confused flour beetle Tribolium confusum Jacquelin du Val, and the yellow mealworm beetle, Tenebrio molitor L. Mortality rates were calculated after 4, 8 and 16 h and 1, 2, 3, 4, 5, 6 and 7 days. The isofuranodiene-based nanoemulsion (NE) exhibited high adulticidal effects against T. molitor, and larvicidal activity against T. castaneum and T. confusum, reaching 98.6, 97.4 and 93.5% at 1000 ppm after 7 days of exposure, respectively. On the other hand, the NE caused low mortality rates on adults of T. castaneum and T. confusum, as well as on larvae of T. molitor, not exceeding 26.2, 10.0 and 15.6% at 1000 ppm, 7 days post-exposure, respectively. Overall, the 3% (w/w) isofuranodiene-based NE developed here has a significant potential to protect stored wheat from tenebrionid beetle infestations.     

Mortality and progeny production of four stored-product insect species on three grain commodities treated with Beauveria bassiana and diatomaceous earths

Laboratory bioassays were conducted to estimate the effectiveness of the entomopathogenic fungus Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae) at 1.5 × 10⁸ and 1.5 × 10¹⁰ conidia/kg grain alone or mixed with the diatomaceous earths (DEs) Protect-It at 150 ppm and DEBBM at 50 ppm against adults of the rusty grain beetle, Cryptolestes ferrugineus Stephens (Coleoptera: Laemophloeidae), the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and the psocid, Liposcelis paeta Pearman (Psocoptera: Liposcelididae). The adult insects were exposed to treated grains for 7, 14 and 21 days. For progeny emergence data, the treated units were remained under the same conditions for 62 days for all beetles and 30 days for psocids. The results revealed significant differences in mortalities of insect species among treatments and grains. The combined use of B. bassiana, at the highest dose, and DE increased adult mortalities of all species. In general, the treatments were least effective against T. castaneum and most effective against L. paeta. More numbers of dead insects were found on wheat than on rice or maize. The emergence of progeny was considerably reduced on grains treated with B. bassiana, at the highest dose, plus DE. The results of the present study suggest that the effectiveness of B. bassiana is fortified by the addition of DEs, but it varies among exposed insect species and commodities.