Phosphine resistance, respiration rate and fitness consequences in stored-product insects

Resistance to fumigants has been frequently reported in insect pests of stored products and is one of the obstacles in controlling these pests. The authors studied phosphine resistance and its physiological basis in adult insects of 12 populations of Tribolium castaneum (Herbst) (Tenebrionidae), ten populations of Rhyzopertha dominica (F.) (Bostrichidae) and eight populations of Oryzaephilus surinamensis L. (Silvanidae) from Brazil, and the possible existence of fitness costs associated with phosphine resistance in the absence of this fumigant. The bioassays for the detection of phosphine resistance followed the FAO standard method. The production of carbon dioxide and the instantaneous rate of population increase (r(i)) of each population of each species were correlated with their resistance ratios at the LC(50). The resistance ratio at LC(50) in T. castaneum ranged from 1.0- to 186.2-fold, in R. dominica from 2.0- to 71.0-fold and in O. surinamensis from 1.9- to 32.2-fold. Ten populations of T. castaneum, nine populations of R. dominica and seven populations of O. surinamensis were resistant to phosphine. In all three species there was significant association (P < 0.05) between respiration rate and phosphine resistance. The populations with lower carbon dioxide production showed a higher resistance ratio, suggesting that the lower respiration rate is the physiological basis of phosphine resistance by reducing the fumigant uptake in the resistant insects. Conversely, populations with higher r(i) showed lower resistance ratios, which could indicate a lower rate of reproduction of the resistant populations compared with susceptible populations. Thus, management strategies based on the interruption of phosphine fumigation may result in reestablishment of susceptibility, and shows good potential for more effective management of phosphine-resistant populations.     

Phosphine-induced walking response of the lesser grain borer (Rhyzopertha dominica)

BACKGROUND: In spite of the intensive worldwide use of phosphine against stored-product insects, its potential sublethal effects on targeted insect species is seldom recognised and may compromise the efficacy of this fumigant, particularly against phosphine-resistant insects. Therefore, the behavioural response of three populations of the lesser grain borer Rhyzopertha dominica (Coleoptera: Bostrichidae) to sublethal phosphine exposure was assessed. RESULTS: Concentration-mortality bioassays with phosphine confirmed the resistance status of the insect populations studied, with levels of phosphine resistance of 40.8-fold and 85.7-fold compared with the susceptible population. Regarding walking behaviour, determined using a digital video-tracking system, the phosphine-resistant populations were less active than the susceptible population. In addition, sublethal phosphine exposure decreased the walking activity of all three populations. CONCLUSION: Phosphine exposure decreased walking ativity. Such reduced walking activity is likely to lower the respiration rate, thereby minimising phosphine uptake. As higher phosphine resistance was also associated with reduced walking activity, phosphine-resistant populations may resist phosphine fumigation even more efficiently on account of this behavioural trait, further aiding their physiological resistance, and should be a matter of concern. Copyright © 2012 Society of Chemical Industry     

Problems Arising from Insecticide Resistance in Storage Pests1

A survey of the occurrence of insecticide or fumigant resistant strains of stored-product insects is given. Studies on the geographic distribution of insecticide-resistant strains show that some resistances are localised and others almost universal. Cross-resistance has been studied in few strains, but malathion resistance may involve resistance to other organo-phosphorus compounds in at least 6 species. Although resistance to pyrethroids and juvenile hormone analogues is known, these compounds show promise for future use against storage pests.     

臭氧对不同磷化氢抗性储粮害虫作用效果研究

为探明臭氧对主要储粮害虫的熏蒸效果及其与磷化氢抗性的关系,本研究采用臭氧发生装置测定浓度为0.84 g/m^3臭氧对5种主要储粮害虫(赤拟谷盗、杂拟谷盗、玉米象、锈赤扁谷盗、谷蠹)的熏蒸效果,及对具有不同磷化氢抗性的赤拟谷盗(抗性系数为1.7~862.7)和杂拟谷盗(抗性系数为2.3~144.7)的毒力效果,并统计分析了试虫经臭氧熏蒸1 h后在不同时间内的行为状态差异。研究结果表明:在0.84 g/m^3臭氧浓度下,5种害虫对臭氧均具有较强敏感性,其LT50在0.58~1.53 h,其中谷蠹对臭氧的耐受性相对最强;赤拟谷盗和杂拟谷盗的不同磷化氢抗性品系对臭氧的敏感性差异较小,表明磷化氢与臭氧无交互抗性。经臭氧熏蒸1 h后,试虫有正常爬行、非正常爬行和死亡3种行为状态,其中较大比例的试虫(24%~66%)处于非正常爬行状态,且持续时间较长,约10 d后此部分试虫恢复正常或死亡。结果表明,臭氧对储粮害虫具有高效性和广谱性,且与磷化氢无交互抗性,可作为储粮害虫防治及磷化氢抗性治理药剂。     

Efficacy of vaporised ethyl formate/carbon dioxide formulation against stored‐grain insects: effect of fumigant concentration,exposure time and two grain temperatures

BACKGROUND: The ethyl formate/carbon dioxide (CO₂) formulation Vapormate? is a rapid‐acting fumigant being developed for the control of stored‐grain insects. The effects have been investigated of concentration, exposure times of 1, 3, 24 and 72 h and two grain temperatures, 15 and 25 °C, on its efficacy against mixed‐stage cultures of Sitophilus oryzae (L.) Tribolium castaneum (Herbst) and strongly phosphine‐resistant Rhyzopertha dominica (F.) strain QRD569. RESULTS: High mortalities (≥92%) of mixed‐stage cultures of all three species were obtained when grain was fumigated with the formulation (193 g m^?3 ethyl formate) for 1 h. Complete control of R. dominica QRD569 and T. castaneum was achieved with 63 and 76 g m^?3 ethyl formate respectively, with exposure for 24 h, whereas mean mortality of S. oryzae was 86% under the same conditions. Mortalities of S. oryzae juvenile stages were significantly lower than adults under the conditions tested, which was due to pronounced tolerance of mid‐stage pupae to the fumigant. Reducing grain temperature from 25 to 15 °C had no effect on insect mortality. CONCLUSION: Ethyl formate/CO₂ formulation is highly effective against stored‐grain insects over a range of concentrations and exposure times. Efficacious fumigations were conducted in as little as 1 h, and a strongly phosphine‐resistant R. dominica strain was readily controlled with the fumigant. Copyright © 2009 CSIRO, Australia. Published by John Wiley & Sons, Ltd     

Improved efficacy of ethyl formate against stored grain insects by combination with carbon dioxide in a 'dynamic' application

Ethyl formate is being evaluated as a fumigant for stored grain as it is a potential alternative to the ozone-depleting fumigant methyl bromide and to phosphine, which is under pressure owing to the development of strong resistance in stored grain insects. However, use of ethyl formate faces significant challenges, such as poor penetration through grain, significant losses to grain sorption, high concentrations of fumigant required to control insects, and flammability risks, which have limited its further development. In this study it was found that the combination of carbon dioxide (5-20%) with ethyl formate significantly enhanced efficacy of the fumigant against external living stages of the rice weevil, Sitophilus oryzae (L.), the lesser grain borer, Rhyzopertha dominica F., and the flour beetle, Tribolium castaneum (Herbst). Dynamic application of ethyl formate and carbon dioxide mixture (100 mg litre-1 ethyl formate, 20% CO2) pumped through a model silo containing wheat (50 kg) for one gas exchange was also investigated. A flow rate of 6 litres min-1 gave a relatively even distribution of fumigant throughout the grain column and similar mortality levels among cultures of S. oryzae and T. castaneum placed at three positions, the top, middle and bottom of the column. Mortality of 99.8% of mixed stage cultures of T. castaneum and 95.1% of S. oryzae was achieved in 3 h exposures to 111 and 185 mg ethyl formate h litre-1 respectively applied by the dynamic method. It is concluded that the combination of carbon dioxide with ethyl formate and dynamic application enhances distribution and efficacy of the fumigant against stored grain insects.     

Effect of exposure period on degree of dominance of phosphine resistance in adults of Rhyzopertha dominica (Coleoptera: Bostrychidae) and Sitophilus oryzae (Coleoptera: Curculionidae)

Degree of dominance of phosphine resistance was investigated in adults of Rhyzopertha dominica F and Sitophilus oryzae L. Efficacy of the grain fumigant phosphine depends on both concentration and exposure period, which raises the possibility that dominance levels vary with exposure period. New and published data were used to test this possibility in adults of R dominica and S oryzae fumigated for periods of up to 144 h. The concentrations required for control of homozygous resistant and susceptible strains and their F1 hybrids decreased with increasing exposure period. For both species the response lines for the homozygous resistant and susceptible strains and their F1 hybrids were parallel. Therefore, neither dominance level nor resistance factor was affected by exposure period. Resistance was incompletely recessive and the level of dominance, calculated at 50% mortality level, was -0.59 for R dominica and -0.65 for S oryzae. The resistant R dominica strain was 30.9 times more resistant than the susceptible strain, compared with 8.9 times for the resistant S oryzae strain. The results suggest that developing discriminating doses for detecting heterozygote adults of either species will be difficult.     

Response of five species of stored-product insects to phosphine in oxygen-deficient atmospheres

Five species of stored-product insects, [Tribolium castaneum (Herbst), T. confusum (DuVal), Rhyzopertha dominica (F.), Sitophilus oryzae (L.) and S. granarius (L.)], exposed to phosphine in nitrogen and various oxygen-deficient atmospheres, showed reduced susceptibility to three concentrations of phosphine (0.02, 0.04 and 0.2 mg litre^?1). The last two species also showed reduced susceptibility to phosphine at 2.0 mg litre^?1. Generally mortality increased progressively as the level of oxygen was raised. There was a close relationship between the ability of insects to withstand anoxia and survival from phosphine treatment.     

Extraction of allyl isothiocyanate from horseradish (Armoracia rusticana) and its fumigant insecticidal activity on four stored‐product pests of paddy

BACKGROUND: Isothiocyanates (ITCs) extracted from Armoracia rusticana Gaertn., May & Scherb. have been shown previously to have insecticidal activity. Allyl isothiocyanate (AITC), a major component of ITCs with high volatility, was therefore extracted using different methods and tested as a fumigant against four major pest species of stored products, maize weevil Sitophilus zeamais (Motsch.), lesser grain borer Rhizopertha dominica (F.), Tribolium ferrugineum (F.) and book louse Liposcelis entomophila (Enderlein). RESULTS: Whereas there was no significant difference between hydrodistillation and supercritical carbon dioxide fluid extraction in extraction rate for AITC from A. rusticana, both methods resulted in higher extraction efficiency than water extraction. AITC fumigation showed strong toxicity to the four species of stored‐product pests. Adult mortality of 100% of all four pest species, recorded after 72 h exposure to AITC fumes at an atmospheric concentration of 3 µg mL^?1, showed no significant difference from that of insects exposed to phosphine at 5 µg mL^?1, the recommended dose for phosphine. CONCLUSIONS: The results suggest good insecticidal efficacy of AITC against the four stored‐product pests, with non‐gaseous residuals on stored products. AITC obtained from A. rusticana may be an alternative to phosphine and methyl bromide against the four pest species. Copyright © 2009 Society of Chemical Industry     

Inhibition of egg development by phosphine in the cosmopolitan pest of stored products Liposcelis bostrychophila (Psocoptera: Liposcelididae)

Phosphine-induced delay in development of eggs was investigated as a mechanism of resistance to this fumigant in Liposcelis bostrychophila Badonnel. One-day-old eggs of a susceptible and a strongly resistant strain of L bostrychophila were exposed to a range of phosphine concentrations for 6days at 30 (+/- 1) degrees C and 70 (+/- 2)% RH. Delay in mean hatching period occurred in both susceptible and resistant eggs, although it was more pronounced in the latter. A maximum delay of 2.65 days was recorded for eggs of the susceptible strain at 0.01 mg litre(-1) (the highest concentration at which eggs survived) and 13.39 days for the resistant strain at 1 mg litre(-1) (the highest concentration tested). Delay in egg development time was positively correlated with increasing phosphine concentration. Our results reveal that the most successful strategy to control resistant L bostrychophila is to apply relatively low concentrations of phosphine for extended exposure times (eg 0.05 mg litre(-1) for 16 days) that allow all eggs to hatch to the much less tolerant nymph stage.     

Ethyl formate plus methyl isothiocyanate--a potential liquid fumigant for stored grains

BACKGROUND: Methyl bromide is being phased out for use on stored commodities, as it is listed as an ozone‐depleting substance, and phosphine is the fumigant widely used on grains. However, phosphine resistance occurs worldwide, and phosphine fumigation requires a long exposure period and temperatures of > 15 °C. There is an urgent requirement for the development of a fumigant that kills insects quickly and for phosphine resistance management. This paper reports on a new fumigant formulation of 95% ethyl formate plus 5% methyl isothiocyanate as an alternative fumigant for stored grains. RESULTS: The formulation is stable for at least 4 months of storage at 45 °C. A laboratory bioassay with the formulation showed that it controlled all stages of Sitophilus oryzae (L.), Sitophilus granarius (L.), Tribolium castaneum (Herbst), Rhyzopertha dominica (F.), Trogoderma variabile Ballion and Callosobruchus maculatus (Fabricius) in infested wheat, barley, oats and peas at 80 mg L^?1 for 5 days, and in canola at both 40 mg L^?1 for 5 days and 80 mg L^?1 for 2 days at 25 ± 2 °C. After an 8–14 day holding period, residues of ethyl formate and methyl isothiocyanate in wheat, barley, peas and canola were below the experimental permit levels of 1.0 and 0.1 mg kg^?1. However, fumigated oats needed an 18 day holding period. CONCLUSIONS: The findings suggest that the ethyl formate plus methyl isothiocyanate formulation has potential as a fumigant for the control of stored‐grain insect pests in various commodities. Copyright © 2011 Society of Chemical Industry     

Toxicity of phosphine to five species of stored-product insects in atmospheres of air and nitrogen

The responses of five species of adult stored-product insects [Tribolium castaneum (Herbst), T. confusum (DuVal), Rhyzopertha dominica (F.), Sitophilus oryzae (L.) and S. granarius (L.)] to various doses of phosphine in atmospheres of air or nitrogen were determined. A few S. oryzae and S. granarius survived phosphine concentrations up to 80 mg litre^?1 for 24 h in normal (air) or nitrogen atmospheres. By contrast, T. castaneum, T. confusum and R. dominica could not tolerate more than 0.2 mg phosphine litre^?1 in air for 12 h. However, they could tolerate up to 10 mg phosphine litre^?1 in nitrogen for that period. Increasing the phosphine concentration resulted in increased mortalities of the last three species. The possible mechanism underlying this response is discussed. The reproductive potential of insects surviving phosphine exposure in a nitrogen atmosphere was unaffected.     

The action of phosphine against the eggs of phosphine-resistant and -susceptible strains of Rhyzopertha dominica F

The effect of phosphine at exposure periods of 24, 48 and 120 h on hatching and mortality of 0- to 1-day-old eggs of susceptible (TN6) and resistant (FC10) strains of Rhyzopertha dominica of field origin was investigated. The fumigant affected hatching in both the strains. In a 48-h exposure at 27 (+/- 2) degrees C, the LD99 doses for the eggs of TN6 and FC10 were 0.56 and 3.25 mg litre-1, respectively. Significant reduction in hatching was observed in treated batches with progressive increase of phosphine dose in the first 2-3 days. On subsequent days the numbers hatching were often similar to those in controls, and sometimes exceeded control hatch, especially following a 5-day exposure. A critical change in the order of susceptibility of egg and adult stages of the two strains was noticed. In 48-h exposures, eggs of the susceptible strain were more tolerant than their adults, whilst the reverse was true in the resistant strain.     

The significance of response time in the detection and measurement of fumigant resistance in insects with special reference to phosphine

The significance of the time of assessment in measurements of fumigant toxicity was examined, using the response of adults of Tribolium castaneum to phosphine. When assessments were made before end-point was reached, large differences were obtained in mortality levels and in parameters of probit lines fitted to the data. These differences reflected the concentration- and dosage-dependent nature of the speed of action, observed previously with this fumigant. The implications of this in the measurement of resistance were examined. In this instance, comparisons at the same concentration were not greatly affected if seven days or longer had elapsed after exposure. However, it was clear that if different concentrations were used for the susceptible strain and the resistant strain, comparisons made before the end-point for acute mortality response was reached could be grossly misleading. This is further discussed in the context of some recent reports of phosphine resistance. The results of this study are of significance in assessing the effect of any poison, drug or other stimulus applied to a group of organisms when the time to respond is likely to vary among individuals.     

Metabolism of methyl bromide by susceptible and resistant strains of the granary weevil, Sitophilus granarius (L.)

Methyl bromide was metabolized by susceptible and resistant strains of adult granary weevil, Sitophilus granarius (L.), mainly by conjugation with glutathione. S-Methyl glutathione and S-methyl cysteine were produced by both strains and S-methyl glutathione sulfoxide was identified as a metabolite in the resistant strain. In the untreated insects, no significant difference was observed in glutathione S-transferase activity but the resistant contained approximately twice as much glutathione per insect as the susceptible strain. When the insects were treated with methyl bromide, the glutathione content of both strains was lowered; proportionally, however, the decrease was considerably higher in the susceptible than in the resistant strain. These results indicate that conjugation of methyl bromide with glutathione is a major detoxication pathway and tolerance to this fumigant is related, in part at least, to the level of glutathione in the granary weevil.     

Insect Growth Regulators and their Potential in the Management of Stored-product Insect Pests

Concern about the impact of pesticides on both health and environment has resulted in the search for alternative control measures for stored-product insect pest. Amongst such alternatives are insect growth regulators (IGRs), a class of biorational compounds that disrupt the normal development of insects. Because of their selectivity of action, these compounds appear to fit the requirements for Third Generation Pesticides, environmentally benign and safer grain protectants. IGRs have been developed commercially and are being used to control insect pests in agriculture, forestry, public health and stored products. IGRs affect the biology of treated insects, for example, both embryonic and post-embryonic development, reproduction, behaviour and mortality. Abnormal morphogenesis is the observed effect of IGR action on the insects. Many of them are more potent than current insecticides, even against the eggs.Compared with the conventional insecticides, IGRs do not exhibit quick knock-down in insects or cause mortality, but the long-term exposure to these compounds largely stops the population growth, as a result of the effects mentioned in both the parents and progeny.The present paper focuses on the previous works on different IGRs available commercially. Also, their possible role in the management of stored-product insect pests has been discussed.     

Influence of phosphine on hatching of Cryptolestes ferrugineus (Coleoptera: Cucujidae), Lasioderma serricorne (Coleoptera: Anobiidae) and Oryzaephilus surinamensis (Coleoptera: Silvanidae)

The hatching and mortality response of 0- to 48-h-old eggs of field strains of the stored-product insects Cryptolestes ferrugineus (Stephens), Lasioderma serricorne (F) and Oryzaephilus surinamensis (L) following phosphine fumigation for 24, 48 or 120 h at 27 (+/- 2) degrees C was investigated. Hatching was delayed and reduced in the first few days in a phosphine-resistant strain of C ferrugineus that was treated with 2.0-7.0 mg litre(-1) doses for 48 h (5-80% mortality) and with 1.0-2.0 mg litre(-1) for 120 h (44-84% mortality). In both the exposures there were significant increases in hatching on later days when compared with the corresponding controls. Developmental delay was, however, not evident in susceptible strains of C ferrugineus, L serricorne and O surinamensis that were exposed to phosphine for 24 h.     

Effect of phosphine dose on sorption in wheat

BACKGROUND: In spite of the extensive use of phosphine fumigation around the world to control insects in stored grain, and the knowledge that grain sorbs phosphine, the effect of concentration on sorption has not been quantified. A laboratory study was undertaken, therefore, to investigate the effect of phosphine dose on sorption in wheat. Wheat was added to glass flasks to achieve filling ratios of 0.25-0.95, and the flasks were sealed and injected with phosphine at 0.1-1.5 mg L(-1) based on flask volume. Phosphine concentration was monitored for 8 days at 25 degrees C and 55% RH. RESULTS: When sorption occurred, phosphine concentration declined with time and was approximately first order, i.e. the data fitted an exponential decay equation. Percentage sorption per day was directly proportional to filling ratio, and was negatively correlated with dose for any given filling ratio. Based on the results, a tenfold increase in dose would result in a halving of the sorption constant and the percentage daily loss. Wheat was less sorptive if it was fumigated for a second time. CONCLUSIONS: The results have implications for the use of phosphine for control of insects in stored wheat. This study shows that dose is a factor that must be considered when trying to understand the impact of sorption on phosphine concentration, and that there appears to be a limit to the capacity of wheat to sorb phosphine.     

Effects of time and concentration on mortality of phosphine-resistant Sitophilus oryzae (L) fumigated with phosphine

The effects of exposure period and phosphine concentration on mortality of susceptible and resistant Sitophilus oryzae (L) were investigated. Although S oryzae is one of the world's most serious pests of stored grain there are few data on the practical significance of phosphine resistance in this species. The strains investigated were an Australian susceptible strain, a homozygous resistant strain exhibiting a level of resistance common in Australia and an unselected field strain from China with a much stronger resistance. Fumigations were carried out at 25 degrees C on adults and mixed-age cultures. For adults of all three strains and mixed-age cultures of the susceptible and resistant Australian strains, the relationship between concentration and time could be described by equations of the form Cnt = k. In all cases n < 1, indicating that time was a more important variable than concentration. In all fumigations of adults the resistant strains were harder to kill than the susceptible strain. However, in fumigations of mixed-age cultures, which contained the tolerant pupal stage, the difference between susceptible and resistant strains was more pronounced at lower concentrations than higher concentrations. For example, at 0.02 mg litre-1 the estimated LT99.9 for mixed-age cultures of the Australian resistant strain (27 days) is 3.4 times that of the susceptible strain (8 days), but at 1 mg litre-1 there is no difference between the two strains (4 days). Limited data on the Chinese resistant strain supported this finding. Twenty-three days exposure at 0.02 mg litre-1 had no effect on mixed-age cultures of this strain, but there were no survivors after 5 days exposure to 1 mg litre-1.     

Changes in phosphine sorption in wheat after storage at two temperatures

BACKGROUND: Wheat can be stored for many months before being fumigated with phosphine to kill insects, so a study was undertaken to investigate whether the sorptive capacity of wheat changes as it ages. Wheat was stored at 15 or 25 °C and 55% RH for up to 5.5 months, and samples were fumigated at intervals to determine sorption. Sealed glass flasks (95% full) were injected with 1.5 mg L^?1 of phosphine based on flask volume. Concentrations were monitored for 11 days beginning 2 h after injection. Some wheat samples were refumigated after a period of ventilation. Several fumigations of wheat were conducted to determine the pattern of sorption during the first 24 h. RESULTS: Phosphine concentration declined exponentially with time from 2 h after injection. Rate of sorption decreased with time spent in storage at either 15 or 25 °C and 55% RH. Rate of sorption tended to be lower when wheat was refumigated, but this could be explained by time in storage rather than by refumigation per se. The data from the 24 h fumigations did not fit a simple exponential decay equation. Instead, there was a rapid decline in the first hour, with phosphine concentration falling much more slowly thereafter. CONCLUSIONS: The results have implications for phosphine fumigation of insects in stored wheat. Both the time wheat has spent in storage and the temperature at which it has been stored are factors that must be considered when trying to understand the impact of sorption on phosphine concentrations in commercial fumigations. Copyright © The state of Queensland (through the Department of Employment, Economic Development and Innovation) 2009. Published by John Wiley and Sons, Ltd.