Field validation of phosphine efficacy on the first recorded resistant strains of Sitophilus granarius and Tribolium castaneum from the Czech Republic

This study first estimated the current state of phosphine sensitivity (using a knock-down/KT₁₀₀/Degesch kit) in Sitophilus granarius (23 strains) and Tribolium castaneum (8 strains) in Czech Republic grain stores. The resistance of S. granarius (21.7% resistant strains; coefficient of resistance KT₁₀₀ ranged from 0.5 to 2.3 among strains) was substantially lower and less frequent than that of T. castaneum (87.5% resistant strains; coefficient of resistance KT₁₀₀ ranged from 0.9 to 52.5 among strains). The phosphine efficacy of the laboratory and field (i.e., resistant) pest strains was validated during commercial fumigation when suboptimal tarpaulin sealing resulted in low-concentration phosphine exposure (Ct products ranged from 5.9 to 7.4 g*hr/m³). Although even low-dose fumigation led to 100% adult mortality of both laboratory and field strains of S. granarius and laboratory strains of T. castaneum, the mortality of the field strain of T. castaneum ranged from 47% to 95%. Larval emergence from the fumigated commodity samples with pest eggs was zero or near zero for laboratory strains, while 1.3–6.0 (S. granarius) and 63.7–80.00 (T. castaneum) field-strain larvae emerged per sample (100 g). This study shows that although a high proportion of the tested pest populations were still sensitive, several T. castaneum populations showed an elevated level of resistance that may decrease field fumigation efficacy, especially under suboptimal phosphine dosage conditions.     

Developmental and population growth rates of phosphine-resistant and -susceptible populations of stored-product insect pests

Phosphine resistance positively contributes towards an individual's fitness under phosphine fumigation. However, phosphine resistance may place resistant individuals at a fitness disadvantage in the absence of this fumigant, which can be exploited to halt or slow down the spread of resistance. This study aimed to determine if there is a fitness cost associated with phosphine resistance in populations of the red flour beetle (Tribolium castaneum (Herbst)), the lesser grain borer (Rhyzopertha dominica (F.)) and the sawtoothed grain beetle (Oryzaephilus surinamensis (L.)). The developmental rate and population growth of phosphine-resistant and -susceptible populations of these three species of stored-product insects were therefore determined under phosphine-free environment. The majority of the phosphine-resistant populations exhibited lower developmental and population growth rates than the susceptible populations indicating that phosphine resistance is associated with fitness cost in all three species, which can potentially compromise the fixation and dispersal of the resistant genotypes. Nonetheless, some phosphine-resistant populations did not show a fitness cost. Therefore, resistance management strategies based on suppression of phosphine use aiming at eventual reestablishment of phosphine susceptibility and subsequent reintroduction of this fumigant will be useful only for insect populations exhibiting a fitness cost associated with phosphine resistance. Therefore recognition of the prevailing phosphine-resistant genotypes in a region is important to direct the management tactics to be adopted.     

Evaluation of dosimeter tubes for monitoring phosphine fumigations

Within integrated pest management options, fumigation of stored products is one method to help control post-harvest insect infestations in our food and agricultural products. Fumigant gas concentration monitoring is important to confirm that the treatment was adequate to achieve the desired insect control, but monitoring can be relatively expensive and labor intensive. This study evaluated how accurately dosimeter tubes could monitor phosphine fumigation treatments. The dosimeter tube is designed to continuously react with phosphine gas during the fumigation period and yields a measurement in terms of concentration 1 time product or CT, which can be interpreted as cumulative exposure. Two models of dosimeter tubes were evaluated (high range and low range). The reference method for these trials were wireless phosphine monitoring sensors, which recorded gas concentrations at hourly intervals during an exposure, and from this a CT product was also calculated. Model LPG-1, high-range dosimeter tube, measured within ± 25% of the phosphine monitoring sensors for CT dosages less the 70,000 ppm1hr. Model LPG-2, low-range tube, tended to significantly over-estimate phosphine CT dosage by 50%–100% of the phosphine monitoring sensor references. Secondly, bioassays of fumigant efficacy were performed using susceptible and resistant adult Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), lesser grain borers, and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), red flour beetle, for estimating insect control at the varied fumigation CT treatments. For the susceptible strains, CT dosages ∼5000 ppm1hr controlled both species. However, the insect control varied from 60% to 100% for resistant adults at CT dosages of ∼20,000 ppm1hr. The dosimeter tubes function in these ranges of dosages where each insect species are controlled and the dosimeter tube model LPG-1 provides reasonable estimates of the fumigation dosage for a given treatment level.     

Phosphine resistance in saw-toothed grain beetle,Oryzaephilus surinamensis in the United States

Phosphine (PH₃) fumigation resulting in sub-lethal exposure has led to the development of phosphine resistance in many stored-product insect species worldwide and is a major challenge to the continued effective use of phosphine. In 2016 phosphine resistance was found in Tribolium castaneum (Herbst) and Plodia interpunctella (Hübner) collected from California dried fruit and nut processing facilities. Although Oryzaephilus surinamensis (L.) infests grain, dried fruit, and nuts in storage and processing facilities, phosphine resistance in this species has not been studied in the United States. In this study, the discriminating dose of phosphine for O. surinamensis eggs was estimated using a laboratory susceptible strain; it was found to be 28.4 ppm over a 72-h fumigation period (1 mg/L of phosphine = 714.18 ppm or 1 ppm = 0.0014 mg/L). Discriminating dose bioassays were used to determine phosphine resistance in both eggs and adults of 14 different populations collected from California and Oklahoma. Resistance to phosphine was detected in four out of 14 populations in adults and nine out of 14 populations in eggs and ranged from 2 to 100%. Phosphine percent survival values in both adults and eggs of three populations, namely, Box BR, Box BF, and OKWat were >90%. Lethal concentration values required to kill 99% of individuals in samples for adults of these three populations were predicted as 320.5, 290.7, and 263 ppm, respectively, and those for eggs were 1030.7, 1055.9, and 564.5 ppm, respectively, over a 72-h fumigation period. This study confirms that phosphine resistance is present in O. surinamensis in the United States.     

Mobility parameters of Tribolium castaneum and Rhyzopertha dominica populations with different susceptibility to phosphine

Phosphine (PH₃) is the most commonly used fumigant to protect stored products from arthropod infestations worldwide. Our knowledge about the behavioral differences between phosphine-resistant and -susceptible stored product pest populations is limited. This study evaluated differences in mobility and behavior of populations of two major stored product insects, Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.), which have different susceptibility to phosphine. In this regard, laboratory bioassays in Petri dish arenas were designed to determine if phosphine resistance has an impact on the walking and mobility behavior of adult beetles of both species. Results indicated that there were significant differences between resistant and susceptible populations for both species. Regarding velocity, R. dominica susceptible individuals moved faster than resistant ones. However, the resistant population showed reduced activity for several parameters tested compared to the susceptible population. Similar trends were also noted for T. castaneum. Knowledge of these parameters should be further utilized in management tactics, as resistant populations may behave in a different way in key management indicators such as trapping and sampling, as compared with susceptible ones.     

Phosphine resistance in Brazilian populations of Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae)

Phosphine resistance was assessed in adults of 22 Brazilian populations of Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae). The concentration-mortality bioassays for the detection of phosphine resistance followed the FAO standard method. Twenty populations of S. zeamais were resistant to phosphine and the resistance ratios at the LC₅₀ ranged from 1.1- to 86.6-fold. This is the first report of phosphine resistance in populations of S. zeamais in Brazil, where previous surveys did not detect resistance in this species. There was significant variation in respiration rate (CO₂ production) among the populations (P < 0.05). Respiration rate was significantly inversely correlated with phosphine resistance for this species (P < 0.05). The populations with lower respiration rates showed higher levels of phosphine resistance, suggesting that the lower respiration rate is associated with the physiological basis of phosphine resistance due to reduced fumigant uptake.     

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

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

Phosphine resistance in stored-product insects collected from various grain storage facilities in Morocco

Despite heavy dependence on phosphine (PH₃) for fumigating stored products, the resistance status of insect pests in Morocco has never undergone a thorough investigation. Some control failures with PH₃ were reported in Morocco, and a previous study showed two field populations of Sitophilus oryzae to be highly resistant to phosphine.We surveyed phosphine resistance in field populations of three major insect pests of stored wheat in Morocco. Around 32% of the samples collected at different storage facilities were found to be infested with one or more species of stored-product beetles. First-generation adult beetles, cultured from the field samples, were subjected to a discriminating dose test for phosphine resistance using an FAO method. The results indicated that, with the exception of one population of S. oryzae, all samples tested contained phosphine-resistant individuals. Treatments at up to 1.8 g m⁻³ of phosphine for 20 h, or at 0.18 g m⁻³ for up to 5 days, indicated that a high degree of resistance was already selected in some of the insect populations.Tests using [³²P]-radiolabelled phosphine showed that the mechanism of resistance in the three insect species tested involved a reduced uptake of the fumigant. The study has highlighted an urgent need for reviewing current fumigation practices in Morocco to ensure effective use of phosphine and avoid further selection of resistance.     

Phosphine fumigation of silo bags

Fumigation with phosphine has the potential to disinfest grain stored in silo bags but only limited research has been conducted on whether phosphine fumigation can be undertaken effectively and safely in this form of storage. Fumigation with phosphine was tested on two (70 m) replicate silo bags each containing 240 t of wheat (9.9 and 9.2% m.c.). The target application rate of phosphine was 1.5 g m⁻³ with a fumigation period of 17 days. Aluminium phosphide tablets were inserted into each bag at ten release points spaced at 7 m intervals starting 3.5 m from either end of the bag. A total of 14 bioassay cages containing mixed age populations of strongly phosphine resistant Rhyzopertha dominica (F.) were inserted into each fumigated silo bag. Complete control of all life stages of R. dominica was achieved at all locations in the fumigated silo bags. Phosphine concentrations at release points increased rapidly and remained high for the duration of the fumigation. Concentrations at midway points were always lower than at the release points but exceeded 215 ppm for ten days. The diffusion coefficient of available phosphine averaged over the first three full days of the fumigation for both fumigated silo bags was 2.8 × 10⁻⁷. Venting the silo bag with an aeration fan reduced the phosphine concentration by 99% after 12 h. Relatively small amounts of phosphine continued to desorb after the venting period. Although grain temperature at the core of the silo bags remained stable at 29 °C for 17 days, grain at the surface of the silo bags fluctuated daily with a mean of 29 °C. The results demonstrate that silo bags can be fumigated with phosphine for complete control of infestations of strongly phosphine resistant R. dominica and potentially other species.     

From immobilization to recovery: Towards the development of a rapid diagnostic indicator for phosphine resistance

The aim of this work was to evaluate the insect mobility patterns of phosphine-resistant and -susceptible adults of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) after exposure to phosphine. Exposure bioassays were carried out at two concentrations, 1000 and 3000 ppm, while adults were observed every 15 min, for a total period of 90 min. During this observation interval, adults were visually classified as active (able to walk normally), partially immobilized (not able to walk, but showing a minimal movement), or completely immobilized (no visible movement). After the observation period, all adults were placed in a phosphine-free environment, and they again were classified as active, partially immobilized or completely immobilized. At 1000 ppm, the majority of adults of the susceptible T. castaneum population were quickly immobilized after a 15 min observation period, while in contrast, the majority of adults of the resistant T. castaneum population were still active after the termination of the 90 min interval. At 3000 ppm, the percentage of immobilized susceptible adults was increased at the 15 min observation period, while the majority of resistant adults were immobilized only after 90 min. In the post-exposure period, the vast majority of the susceptible adults were dead. In contrast, most resistant adults recovered, regardless of the concentration that had been exposed. The results of this study delineate major differences in movement in phosphine-resistant and -susceptible T. castaneum strains, and can be applied as a quick diagnostic bioassay for the evaluation of resistance to phosphine in stored product insects.     

Effect of diet on phosphine toxicity,rate of development and reproduction of the rice weevil Sitophilus oryzae (Linnaeus)

One of the loci responsible for strong phosphine resistance encodes dihydrolipoamide dehydrogenase (DLD). The strong co-incidence of enzyme complexes that contain DLD, and enzymes that require thiamine as a cofactor, motivated us to test whether the thiamine deficiency of polished white rice could influence the efficacy of phosphine fumigation against insect pests of stored grain. Three strains of Sitophilus oryzae (susceptible, weak and strong resistance) were cultured on white rice (thiamine deficient), brown rice or whole wheat. As thiamine is an essential nutrient, we firstly evaluated the effect of white rice on developmental rate and fecundity and found that both were detrimentally affected by this diet. The mean time to reach adult stage for the three strains ranged from 40 to 43 days on brown rice and 50–52 days on white rice. The mean number of offspring for the three strains ranged from 7.7 to 10.3 per female over a three day period on brown rice and 2.1 to 2.6 on white rice. Growth and reproduction on wheat was similar to that on brown rice except that the strongly resistant strain showed a tendency toward reduced fecundity on wheat. The susceptible strain exhibited a modest increase in tolerance to phosphine on white rice as expected if thiamine deficiency could mimic the effect of the dld resistance mutation at the rph2 locus. The strongly resistant strain did not respond to thiamine deficiency, but this was expected as these insects are already strongly resistant. We failed, however, to observe the expected synergistic increase in resistance due to combining thiamine deficiency with the weakly resistant strain. The lack of interaction between thiamine content of the diet and the resistance genotype in determining the phosphine resistance phenotype suggests that the mode of inhibition of the complexes is a critical determinant of resistance.     

Genetic diversity and its geographic structure in Sitophilus oryzae (Coleoptera; Curculionidae) across India – implications for managing phosphine resistance

The rice weevil, Sitophilus oryzae, is a serious global pest of stored grains. Fumigation with phosphine gas is the primary control method for S. oryzae, but the indiscriminate and prolonged use of phosphine gas has led to the development of heritable resistance. Developing and implementing an effective phosphine resistance management strategy for S. oryzae relies on an understanding of its genetic diversity and any structuring of that diversity geographically. We therefore sequenced the mitochondrial cytochrome c oxidase subunit I gene from 143 S. oryzae specimens collected from 37 locations across India, and from that assessed the genetic diversity of the species and its phylogeographic structuring. In addition, we compared the genetic diversity in Indian S. oryzae populations (the hypothesised origin of this beetle) to global populations. Genetic diversity was low in Indian S. oryzae, with only eight haplotypes (including two very common haplotypes) identified. The low level of mitochondrial diversity observed in this species appears typical of stored product pests, perhaps suggesting that low mitochondrial diversity is associated with repeated phosphine fumigations, which may eliminate low frequency haplotypes. The genetic diversity of S. oryzae in India is, however, higher than in many other countries, though comparable levels were identified in China. There was no evidence of population genetic structure across India, with most haplotypes found in three of the broad biogeographic regions. This lack of phylogeographic structuring indicates significant gene-flow across India, most likely through the incidental anthropogenic transport of this relatively poor (or reluctant) flyer. The major practical implication is that phosphine resistance management for S. oryzae needs to be dealt with country wide, as populations are not isolated.     

Biochemical mechanisms conferring cross-resistance to fumigant toxicities of essential oils in a chlorpyrifos-methyl resistant strain of Oryzaephilus surinamensis L. (Coleoptera: Silvanidae)

The fumigant toxicities of lavender and ylang-ylang essential oils were tested against a chlorpyrifos-methyl resistant strain (QVOS102) and an insecticide-susceptible reference strain (VOS48) of the saw-toothed grain beetle, Oryzaephilus surinamensis L. (Coleoptera: Silvanidae). The resistant strain showed 1.3- and 1.6-fold higher tolerance against lavender and ylang-ylang fumigation toxicity, respectively, relative to the susceptible strain. LT₅₀ values calculated as the time to attain 50% mortality of tested insects during fumigation were determined at two different concentrations. At 15 μl/l of air, QVOS102 had 2.9- and 1.4-fold higher LT₅₀ values for lavender and ylang-ylang fumigation toxicity, respectively, than VOS48. At 200 μl/l air, QVOS102 had 6.4- and 2.9-fold higher LT₅₀ values for lavender and ylang-ylang fumigation toxicity, respectively, than VOS48. Piperonyl butoxide, a potential inhibitor of cytochrome P450-dependent monooxygenase, increased fumigant toxicities of the two essential oils against QVOS102. The enhanced tolerance for the essential oil may have resulted from the enhancement of detoxifying enzymes associated with insecticide resistance.     

Allyl isothiocyanate actions on populations of Sitophilus zeamais resistant to phosphine: Toxicity,emergence inhibition and repellency

The risks associated with the use of synthetic insecticides have caused increased interest in the research of essential oils and their main constituents for use in the pest management of stored products. Allyl isothiocyanate (AITC) is the main component of mustard essential oil and has been reported as a potential replacement pesticide for conventional insecticides that control stored product insect pests. Here, we assessed the toxicity (including emergence inhibition) and repellent actions of AITC on Brazilian populations of the maize weevil Sitophilus zeamais (Coleoptera: Curculionidae) resistant to conventional insecticides (e.g., phosphine). We also evaluated physiological (e.g., respiration) and behavioral (e.g., walking and flight) traits of AITC-exposed insects. The AITC showed consistent insecticidal activity against the populations resistant to phosphine and other synthetic insecticides, with LC₅₀ values ranging from 1.5 to 2.9 μL L⁻¹. Significant inhibition of the offspring emergence was achieved after the exposure of parental adults to sublethal levels (i.e., LC₁ and LC₅) of AITC. Reductions in respiration rates were also registered in all the populations sublethally exposed to AITC. In all five populations, a high number of insects avoided AITC-treated (1.5 μL L⁻¹) grain masses, and although individuals of a phosphine-susceptible (i.e., Abre Campo) population increased walking and reduced flight activities, individuals of another phosphine-susceptible (i.e., Tunápolis) population exhibited higher flight activity under AITC exposure. Thus, our findings suggest that AITC is a potential tool that may be integrated into the control strategies of maize weevils where resistance to phosphine and other conventional insecticides is a problem.     

Comparison of efficacy of nitric oxide fumigation under nitrogen and carbon dioxide atmospheres in controlling granary weevil (Sitophilus granaries) and confused flour beetle (Tribolium confusum)

Nitric oxide (NO) was a recently discovered fumigant for postharvest pest control. Because NO reacts with oxygen (O₂) spontaneously to form nitrogen dioxide (NO₂), NO fumigation must be conducted under ultralow oxygen (ULO) atmospheres to preserve NO and nitrogen (N₂) has been used to establish ULO atmospheres in NO fumigation studies in the past. However, carbon dioxide (CO₂) can also be used to ULO atmospheres and CO₂ fumigation was also reported to be effective in controlling certain insect pests and enhancing toxicity of some fumigants. In the present study, NO fumigations under ULO conditions established with N₂ and CO₂ were compared for effects against granary weevil, Sitophilus granaries, and confused flour beetle, Tribolium confusum. All life stages of the two insects were subjected to 12 h fumigation treatments with 0.5–1.0% NO at 25 °C under ULO established with CO₂ (NO–CO₂ treatments) and N₂ (NO–N₂ treatments). The most tolerant life stage for each species in each fumigation treatment was then fumigated with NO for 24 h fumigation at 25 °C to determine an effective treatment. There were no significant differences in mortalities of adults, larvae, and pupae at all NO concentrations between ULO conditions established with CO₂ and N₂ for either insect. NO–CO₂ was, however, significantly less effective than NO–N₂ against eggs of both species. Granary weevil pupae and confused flour beetle eggs were the most tolerant stages for each respective species to NO fumigation: >99% mortality of granary weevil pupae and confused flour beetle eggs were, however, achieved in the 24 h fumigation with 2% and 1% NO, respectively. This study showed that CO₂ can be used to establish ULO atmospheres for NO fumigation, but did not significantly contribute to insect mortality.     

Detection of phosphine resistance in major stored-product insects in Greece and evaluation of a field resistance test kit

The use of phosphine has been effective against a wide range of stored-product pests in different types of commodities and facilities. However, its continuous and improper use has led to resistance development in -several major insect species. Although phosphine resistance has been reported from many countries across the globe, reports from Europe have been very limited. In the present study, we determined phosphine resistance in insect populations that had been collected from a range of storages across Greece, using two different diagnostic protocols. Apart from the traditional Food and Agriculture Organization (FAO) protocol, a field test kit (known as the Detia Degesch Tolerance Test Kit, DDTTPK) was utilized, for “same day” determination of the resistance status of field collected insects. In total, 53 populations belonging to Rhyzopertha dominica, Sitophilus oryzae, Sitophilus granarius, Cryptolestes ferrugineus, Tribolium confusum, Tribolium castaneum and Oryzaephilus surinamensis were tested. For the majority of the species and populations tested, both FAO and DDPTTK provided similar results, for the susceptibility to phosphine and thus, the quick test could be used with success for an initial same day screening of phosphine resistance. Among the tested species, the populations recorded with the most frequent survival at the FAO testing dose of phosphine was that of R. dominica. The dissimilar evaluation and characterization of resistance to phosphine between diagnostic protocols is particularly important, as it poses risks in the over or underestimation of the resistance status of a given population. Our data indicate that the DDPTTK could be used to determine resistance to phosphine in the field, before the initiation of fumigations to disinfest stored commodities.     

Control failure and insecticides resistance in populations of Rhyzopertha dominica (Coleoptera: Bostrichidae) from Colombia

The lesser grain borer Rhyzopertha dominica is a serious pest that causes significant economic damages in stored grains. The presence of this pest is mainly controlled using a small number of insecticides increasing risks of compromising the control efficacy due to the selection of resistant populations. Here, the control failure and potential resistance to pyrethroid and organophosphate insecticides frequently used for the control of R. dominica in Colombia were assessed. Concentration–mortality bioassays with synergized and unsynergized insecticides were used to assess potential metabolic-based pyrethroids resistance mechanisms. Based on the field recommended doses, the results revealed that while the pirimiphos-methyl is still efficient against R. dominica, both type I (i.e., bifenthrin) and type II (i.e., deltamethrin) pyrethroids presented control failure and their insecticidal activity was compromised. Although no differences in susceptibilities between the populations were found for the bifenthrin insecticide, the LC₅₀s of Ibagué (1.83 μl/gr of grains) and El Espinal (1.67 μl/gr of grains) populations were higher than the recommended rate. Furthermore, significant differences in susceptibility to deltamethrin resulted in resistance ratios RR₅₀ > 32-fold and RR₉₅ > 188-fold between the least (El Espinal) and the most (Ibagué) susceptible populations. Moreover, when synergized, the toxicity of deltamethrin was enhanced but resistance ratios kept at high levels (RR₅₀ > 11-fold and RR₉₅ > 40-fold) indicating possible existence of simultaneous metabolic and altered action-site mechanisms of resistance to pyrethroids in Colombian populations. Our results highlight the urgent need for nationwide monitoring of insecticides control failure and resistance in stored pests for better management schemes.     

Efficacy of heat treatment on phosphine resistant and susceptible populations of stored product insects

In this study we evaluated the efficacy of heat treatment on phosphine resistant and susceptible populations of stored product insects at twenty three different commercial facilities in Greece. Heat treatments were carried out by using special equipment, such as Therminate, TempAir and ThermoNox, applied alone or in combination. The overall temperature range was between 33 and 55 °C and the duration of the heat treatments was between 20 and 39 h. Adults of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), and the sawtoothed grain beetle, Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae), were used in the experiments. The field populations were collected from different storage facilities in Greece and were characterized as resistant populations by using the Detia Degesch Phosphine Tolerance Test Kit. Insect mortality was measured at the termination of each trial. Then, the vials were kept in incubator chambers at 25 °C and 65% relative humidity and 65 d later the progeny production was measured in the treated substrate. In light of our findings, in the vast majority of the cases, complete control was observed for both resistant and susceptible populations at all facilities. In general, in the few cases where survival and progeny production was recorded, there was no specific trend towards specific species or population. Overall, based on the current results, heat treatment can be used by the industry as an alternative method for the control of phosphine-resistant adults of R. dominica and O. surinamensis.     

Using immobilization as a quick diagnostic indicator for resistance to phosphine

In the present study, we evaluated a quick diagnostic test for resistance to phosphine in stored-product beetle species. We collected different populations of thirteen species, obtained from different laboratories in different counties, i.e., USA, Greece, Australia, Germany and Spain. There were also tested populations that have been sampled from different facilities (field populations). We used the Detia Degesch Phosphine Tolerance Test Kit (DDPTTK), which is based on the exposure of the insects on a high concentration of phosphine for shorter exposure periods. The tested concentrations to phosphine were 1000 and 3000 ppm. Briefly, 20 adults of the tested populations were placed in a 100 ml plastic syringe. The observations were taken every 2 min and the exposed adults were classified as either walking normally or being immobilized (knocked down), i.e., not walking normally. In light of our findings, the time to reach knockdown of all adults was notably reduced at 3000 ppm in comparison with 1000 ppm. For the majority of the species and laboratory populations tested, at 3000 ppm, the time required for knockdown ranged between 8 and 14 min. In contrast, for some of the field populations, knockdown did not reach 100% even after 300 min of exposure, at either 1000 or 3000 ppm. Based on the results of the present study, we recommend that the DDPTTK can be operated at 3000 ppm, and we provide the critical threshold times per species for the characterization of tolerance/resistance.     

Fumigant toxicity of allyl isothiocyanate to populations of the red flour beetle Tribolium castaneum

The phasing out of methyl bromide as a fumigant, the phosphine resistance problems in stored product insect-pests, and the ever-growing concerns with human health and environmental safety have been guiding the search of alternative fumigants. Allyl isothiocyanate (AITC) is the main component of mustard oil with reported pesticide activity and potential as a fumigant of stored foodstuffs. The fumigant toxicity of AITC was assessed in adults of 18 populations of the red flour beetle Tribolium castaneum (Coleoptera: Tenebronidae). These populations were all susceptible to AITC with negligible variation among them. Two of these populations were further used to test the AITC susceptibility of eggs, larvae (early and late instars), and pupae of T. castaneum. All of the developmental stages of both populations were similarly susceptible to AITC. No cross-resistance between phosphine and AITC was observed. Despite the significant variation in body mass, respiration rate, and fitness among the populations of T. castaneum, they were not correlated with AITC susceptibility. Larvae and adult malformations were observed when larvae and pupae were exposed to AITC. These results show the potential of AICT as an alternative fumigant against stored product insects.