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Trogoderma granarium Everts, the khapra beetle, is a serious pest of stored products throughout the world. Larvae pose a significant threat to stored products because they feed on >100 different commodities, possess the ability to enter facultative diapause, and are difficult to detect. Control methods for T. granarium include fumigation, contact insecticides, trapping, and insecticide-incorporated packaging. The objective of this study was to determine the residual efficacy of two insecticide formulations (methoprene + deltamethrin + piperonyl butoxide synergist Gravista® and methoprene + deltamethrin, DiaconIGR®Plus). These insecticides were evaluated on three stored product commodities, corn, wheat, and brown rice, by exposing T. granarium larvae during a 12-month testing period. Both formulations significantly reduced adult emergence on corn and wheat for 12 months and on brown rice for up to 6 months. Adult emergence was highest at month 12 for corn (8.41%), and brown rice (85.88%), and month 9 for wheat (39.52%), treated with DiaconIGR®Plus or Gravista®, respectively. A biological index used to measure the development of exposed larvae on the treated grain from the larval stage (low values) to adult emergence (high values) was lower (fewer adults) on corn and wheat compared to controls. Despite differences in formulations, each of these grain protectants could be utilized by stored commodity managers to protect commodities during storage and transportation for T. granarium when and if this pest is detected at USA ports of entry.

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BACKGROUND: Sublethal effects of insecticides may negatively affect several biological and behavioral traits of insects. The lethal effects of pirimiphos-methyl and chlorfenapyr have been previously showed on Trogoderma granarium, but little knowledge is available about their sublethal effects at low concentrations on both sexes. Herein, the sublethal effects of pirimiphos-methyl and chlorfenapyr on the mobility of T. granarium males and females were investigated. RESULTS: Lethal concentration (LC) values of pirimiphos-methyl and chlorfenapyr were lower for T. granarium females than males. LC values on males were LC10 = 0.000788 and 0.00139 mg active ingredient (a.i.) cm-2 , LC30 = 0.00350 and 0.00535 mg a.i. cm-2 , and LC50 = 0.00986 and 0.0136 mg a.i. cm-2 for pirimiphos-methyl and chlorfenapyr respectively. LC on females were LC10 = 0.000704 and 0.00110 mg a.i. cm-2 , LC30 = 0.00323 and 0.00428 mg a.i. cm-2 , and LC50 = 0.00925 and 0.0110 mg a.i. cm-2 for pirimiphos-methyl and chlorfenapyr respectively. The walking duration of beetles exposed to LC30 of pirimiphos-methyl was significantly lower than the individuals exposed to LC10 and LC30 of both insecticides and control ones. Pirimiphos-methyl LC30 -exposed males remained more time on their back (101.7 s) than females (46.9 s), while the latter stayed immobile longer than males (381.7 s versus 371.9 s). The highest speed was recorded for control beetles (14.17 mm s-1 females vs. 12.44 mm s-1 males), while the lowest speed was observed in pirimiphos-methyl LC30 -treated males (8.36 mm s-1 ) and females (9.66 mm s-1 ). CONCLUSIONS: Overall, males and females exposed to low concentrations of pirimiphos-methyl and chlorfenapyr showed reduced motility. This knowledge can be exploited further to unlock behavioral effects of insecticides for effective pest management programs in warehouses. © 2023 Society of Chemical Industry.

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BACKGROUND: Khapra beetle (Trogoderma granarium Everts), one of the most important quarantine pests globally, is capable of causing severe infestation and huge economic loss to stored grain, and its interception rate has increased in major global trade countries over the past few years. However, difficulties remain in distinguishing this species with similar ones. In order to assist border ports and warehouses in khapra beetle's effective rapid identification as well as pest control at the early stages of monitoring or interception, we herein developed a new and rapid visual detection assay for T. granarium based on recombinase polymerase amplification (RPA) and the CRISPR/Cas12a system. RESULTS: We designed and selected the first khapra beetle-specific RPA primers and crRNA, and optimized the visualization reaction system (Cas12a/CrRNA = 100 nM/500 nM). With only a 37 °C-heat-source and a blue light torch, RPA and CRISPR/CAS12a-based visualization assays can be completed within 40 min to differentiate between khapra beetle and nine similar Dermestidae species. After DNA extraction using a kit (4-5 h) or a simple method (5 min), the specific amplicons were obtained after a 15 min RPA reaction at 37 °C, followed by a 15 min color reaction under 37 °C in dark conditions using a CRISPR/CAS12a system and a fluorescent probe (5'-FAM/3'-BHQ1 labeled). This method is ingenious to low levels of DNA (10-1 ng µL-1 ) and meets the sensitivity requirements for detecting a single khapra beetle's egg (≈0.7 mm). CONCLUSION: Our specificity and sensitivity analysis inferred that the present visualization system is effective to quickly and uniquely detect khapra beetle at room temperature (37 °C), thereby preventing this species before they spread widely. Our study is suitable for being pushed forward in storage pest management, and provides value as a reference for monitoring and identification of other pests. © 2023 Society of Chemical Industry.

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BACKGROUND: Khapra beetle (Dermestidae: Trogoderma granarium Everts, 1898) is an internationally significant pest of grain crops and stored grain products. Wheat germ traps, routinely used in surveillance sampling of Khapra beetle provide feed-substrates used by the pest throughout its life cycle. However, Khapra beetle larvae, eggs and other traces of the pest, such as larval frass and exuviae, in wheat germ traps are difficult to sort and taxonomically identify. Additionally, high levels of polysaccharides in wheat germ can inhibit PCR based molecular detection of this pest captured in the traps. METHODS AND RESULTS: We have developed a sensitive and low-cost protocol for extracting trace levels of Khapra beetle DNA from an entire wheat germ trap. Overnight digestion of entire trap contents in 6 mL of ATL buffer, followed by a 40 min lysis step was optimal for DNA extraction. Paired with reported qPCR assays, this protocol allows the detection of a few hairs of T. granarium in a typical 2-gram wheat germ trap. CONCLUSION: This DNA extraction protocol makes it possible to perform a more rapid identification of the pest following wheat germ sample collection. The protocol has potential to improve international efforts for Khapra beetle surveillance.

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We evaluated the relative population growth of two stored-product insect species in the genus Trogoderma, the khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae), and the warehouse beetle, Trogoderma variabile Ballion (Coleoptera: Dermestidae). Ten adults of each species were placed in vials containing wheat or paddy rice. These tests were performed at 27 and 32°C and the number of adults in the vials were counted after 35 and 70 days. For all the time intervals and temperatures of both species on wheat, the resulting larval abundances were similar, with the exception of 27°C at 70 days where more T. variabile larvae developed. At the higher temperature, both species had similar population growth on rice. However, the success was mixed at 27°C with T. granarium having a greater abundance after 35 days, while T. variabile dominated after 70 days. Frass production in both commodities was usually similar for the two species, but greater frass production occurred by T. variabile on wheat after 70 days at 27°C, while T. granarium produced more frass on rice after 35 days at 32°C. Both species nearly always caused equivalent commodity damage. Our research suggests that under ideal conditions these two closely, but very differently treated species in commerce from the genus Trogoderma have comparable population growth, and cause similar damage on wheat and rice.

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Khapra beetle, Trogoderma granarium Everts, is one of the economically important quarantine pests that mainly feeds on food grain and proteinaceous materials. Its total development time lasts approximately 40-45 d under favorable environmental conditions. Extreme temperatures, high relative humidity (RH), high larval densities, or low food quality can induce a larval diapause, where the insect can survive for up to a few years, occasionally feeding and molting. Ecological modeling is a helpful tool to study the population dynamics of biological systems. Physi-Biological age method is based on temperature-driven development rate, and factors such as RH and food quality were considered as multipliers. The objective of this study was to develop mathematical models to calculate the survival and development of adults, eggs, larvae, pupae, and oviposition and diapause under different environmental conditions such as temperature, RH, and food quality. Algorithms were developed to simulate the population dynamics for each day and coded in C++. The developed models were validated against the literature data and evaluated using linear regression, R2, and MSE. Population dynamics were simulated under Canadian grain storage conditions, and the developed models predicted that the diapausing larvae survived the extremely cold conditions found in Canadian grain. In contrast, other stages did not survive. The surviving larvae developed to pupae and adults, and females began laying eggs once the temperature became warmer in the grain bins.

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BACKGROUND: Khapra beetle (Trogoderma granarium Everts) is a significant pest of food products around the world, causing great losses of stored grain and produce, with export restrictions imposed on countries with established beetle populations. Khapra beetle is a high-priority exotic invertebrate pest in many countries requiring a rapid quarantine/biosecurity response when incursions occur. To address this, we developed a novel Khapra LAMP (loop-mediated isothermal amplification) assay using a portable real-time fluorometer and an additional 18S ribosomal DNA (18S) insect control LAMP assay for confirmation of the presence of insect DNA. Both LAMP tests can be performed either in a portable real-time fluorometer or using simple, visual colorimetric technique. RESULTS: Both the Khapra and 18S LAMP tests amplify positive samples within ≤ 25 min, with an anneal derivative temperature of 77.7 ± 0.7 °C for Khapra LAMP test and 88.0 ± 1.0 °C for 18S. The new Khapra LAMP assay is sensitive to very low levels of DNA (1.02 × 10-6  ng µL-1 ). Additionally, we developed a gBlock double stranded DNA fragment for use as positive Khapra control with a different anneal derivative of 80 °C. Both assays are simple to use in the field and are capable of amplifying DNA from target beetles, even when samples are partially degraded which is typically found during surveillance activities. By screening a broad panel of Dermestidae species we demonstrate that our new assay is species-specific, with no detections of false positives. Also, we evaluated multiple DNA extraction methods, with both QuickExtract and HotSHOT extraction methods proving suitable for in-field use. CONCLUSION: The novel Khapra and 18S LAMP assays should improve speed, accuracy and confidence of detection of Khapra beetle at incursion points and aid rapid biosecurity responses in any country affected, especially as the assays described here are portable and easy to implement in the field conditions where resources are limited. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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The khapra beetle, Trogoderma granarium Everts, is defined as one of the most important quarantine pests globally, and fumigation with methyl bromide, an ozone-depleting substance, is a common phytosanitary measure currently used. The modified atmosphere (MA), irradiation, and their combination treatments of T. granarium larvae and adults were performed at room temperature (24-26 ℃) to develop an ecofriendly phytosanitary disinfestation measure and to shorten the exposure time and overcome treatment disadvantages of irradiation. Late-stage larvae are determined as the most tolerant stage resulted in large LT99.9968 values of 32.6 (29.2-37.5) and 38.0 (35.1-41.7) days treated under 1% and 2% O2 (with N2 balance) atmosphere, respectively. Ionizing radiation was used to enhance the effect of MA and the mortality was highly significantly affected by all the interaction effects, indicating that the synergistic effects present in all the combined treatments. The synergistic ratios, which is defined as the estimated lethal time for MA treatment (LD90, LD99, and LD99.9968), divided by that of combined treatment, were between 1.47 and 2.47. In the confirmatory tests, no individuals recovered from a sum of 111,366 late-stage larvae treated under 1% O2 atmosphere for 14- or 15-d after 200 Gy irradiation, which resulted in validating the probit estimations and achieving an efficacy of 99.9973% mortality at 95% confidence level. Therefore, these treatment schedules are recommended to disinfest T. granarium infecting commodities for phytosanitary purposes under the warehouse, MA packaging, or in combination with international transportation by train or sea container.

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In Pakistan, the control of stored-product insect pests mainly relies on the use of phosphine gas along with other control tactics. The aim of this study was to determine the level of phosphine resistance among ten differently located populations of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), the granary weevil, Sitophilus granarius (L.) (Coleoptera: Curculionidae), the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and the khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae). Laboratory-susceptible populations of all insect species were also considered in the experiments. Concentration-response bioassays were conducted for each species. All of the tested populations (10 out of 10) of each species were found to be resistant to phosphine, but varied in their level of resistance. Probit analysis estimated LC50 at 2.85, 1.90, 2.54 and 2.01 ppm for laboratory-susceptible populations of R. dominica, S. granarius, T. castaneum and T. granarium, respectively. Against R. dominica, the highest and lowest resistance levels were observed in the Rahim Yar Khan (LC50 at 360.90 ppm) and Rawalpindi (LC50 at 210.98 ppm) populations, respectively. These resistant populations were 126.67- and 74.02-fold more resistant than the laboratory population. The Multan and Lahore populations of S. granarius exhibited the maximum (LC50 at 122.81 ppm) and minimum (LC50 at 45.96 ppm) resistance levels, respectively, i.e., they were 64.63- and 24.18-fold more resistant than the laboratory population. The Layyah population of T. castaneum showed the maximum resistance level (LC50 at 305.89 ppm) while the lowest was observed in the Lahore population (LC50 at 186.52 ppm), corresponding to 120.42- and 73.43-fold more resistant than the laboratory population, respectively. Regarding T. granarium, the Layyah population showed the maximum resistance level (LC50 at 169.99 ppm) while the Lahore population showed the minimum resistance (LC50 at 74.50 ppm), i.e., they were 84.57- and 37.06-fold more resistant than the laboratory population, respectively. Overall, R. dominica presented the highest resistance level, followed by T. castaneum, T. granarium and S. granarius. The current study suggests that the application of phosphine may not be an adequate control strategy for the management of the above tested insect pests in Pakistan.

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The botanical substances constitute valuable alternatives to synthetic insecticides. In the last decades, numerous substances of natural origin have been tested against stored-product insects, mostly as fumigants or for contact toxicity, while there is limited knowledge on the efficacy of plant secondary metabolites if used as grain protectants. In the present study, we evaluated the lethal activity of 2-undecanone, acetic acid, trans-anethole, furfural, (E)-2-decenal and (E, E)-2,4-decadienal as wheat protectants for the management of larvae and adults of two important storage pests, Tenebrio molitor (Coleoptera: Tenebrionidae) and Trogoderma granarium (Coleoptera: Dermestidae). 2-undecanone caused 98.9% mortality to the exposed T. molitor adults at 1000 µl/kg wheat 7 days post-exposure, while acetic acid and furfural followed providing 94.4% and 92.2% mortality respectively. 2-Undecanone and (E)-2-decenal caused the highest mortalities to T. molitor larvae (i.e., 87.8% and 80.0% respectively) exposed to 1000 µl/kg wheat for 7 days. All T. granarium adults were dead at 1000 µl (E)-2-decenal or acetic acid/kg wheat 5 or 7 days post-exposure respectively. Complete (100%) mortality was assessed for larvae exposed to (E, E)-2,4-decadienal and (E)-2-decenal at 1000 µl/kg wheat after 4 and 6 days respectively. Our findings report for the first time that 2-undecanone, (E)-2-decenal, and (E, E)-2,4-decadienal are effective new candidate control agents of different developmental stages of T. molitor and T. granarium.

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A series of tests was performed to examine the development ability of Trogoderma granarium Everts on six different sorghum milling fractions: bran, shorts, coarse grits, fine grits, red dogs, and flour. In the first bioassay, 20 adults and 20 g of each fraction were placed in vials of 60 ml at 30°C and 60% relative humidity for a period of 65 d. In the second part of the experiment, a neonate larva (1-d old) was placed in vials with 1 g of each fraction and exposed at 25, 30, and 32°C until adult emergence. Significantly more live larvae were found in bran compared with flour and to the two grain controls (soft wheat and whole wheat flour), where most dead adults were recorded. The increase in temperature reduced both the time to adult emergence and larval mortality before the appearance of the first adult. Developmental times ranged between 25.5 and 37.5 d, and 23.2 and 29.0 d for 30 and 32°C, respectively. At 25°C, larvae in the milling fractions did not reach 100% adult emergence even after almost 3 mo. However, the first adult appeared in the shorts fraction at all temperatures tested. Our results show that all sorghum milling fractions support the development of T. granarium and that the optimal temperatures for growth and development are 30 and 32°C. These results also have important implications for phytosanitary regulations and international trade, given the quarantine restrictions on this insect.

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Long-lasting insecticide treated netting (LLIN) has a number of potential uses for the control of insect pests. Using such netting, stored products may be protected from insects including the khapra beetle (Trogoderma granarium Everts, Coleoptera: Dermestidae) a widespread pest of many agricultural commodities. Here we first examined whether brief exposures of larvae to LLIN, for less than 30 min, decreased the chance of eventual adult emergence compared to larvae exposed on untreated netting. Next, we observed the responses of larvae that were either not exposed to any netting, exposed to untreated netting, or exposed to LLIN for 10 min and then placed in a wind tunnel and monitored for movement toward a stimulus. The wind-tunnel assay was performed either with or without a lure containing kairomones and pheromones known to be attractive to larvae of this species. There was little effect of the LLIN on adult emergence of exposed larvae. However, there were interacting effects of untreated netting and LLIN relating to thigmotaxis and anemotaxis. Larvae not exposed to netting showed increased likelihood of walking upwind if the semiochemical lure was provided, as expected. A similar pattern was observed when the untreated netting was used, but the larvae became more likely to remain stationary in the assay after acclimating to the net. When LLIN was used, the larvae became more likely to move and there was a baseline increase in the likelihood of moving upwind. However, upwind walking was no longer related to semiochemical presentation. These observations suggest that particular care should be used in relation to the airflow patterns and semiochemical landscape of the warehouse settings in which LLIN is deployed.

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The khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae), is one of the most destructive insect species of stored food worldwide and is subjected to strict phytosanitary legislations. In the present study, we evaluated the egg hatching and larval mortality of T. granarium on concrete surfaces treated with six insecticides (i.e., α-cypermethrin, chlorfenapyr, deltamethrin, pirimiphos-methyl, pyriproxyfen, and s-methoprene) that are registered in Greece for surface treatment and exhibit a broad spectrum of different modes of action. Furthermore, we investigated the influence of the presence of food on egg hatching and larval mortality. Egg hatchability on treated concrete was higher in tests with the presence of food for all tested insecticides, with the exception of s-methoprene. In contrast, larval mortality was lower in treatments where there was nourishment for all insecticides. No egg hatching was recorded on concrete treated with pirimiphos-methyl where there was no food, while with the addition of food, the egg hatching did not exceeded 26.7% after 6 days of exposure. The highest percentage of hatched eggs was recorded on concrete treated with chlorfenapyr (87.7% with food vs. 76.7% without food), followed by deltamethrin (76.7% with food vs. 63.3% without food), pyriproxyfen (50.0% with food vs. 42.2% without food), and α-cypermethrin (28.9% with food vs. 6.7% without food). In the case of s-methoprene, more eggs were hatched in the absence of food (91.1%) in contrast to in the presence of food (86.7%). Regarding mortality, all larvae were dead after 5 days of exposure on pirimiphos-methyl-treated concrete with food. Furthermore, larvae died faster in treatments without food. For α-cypermethrin, 100% mortality was recorded after 4 days of exposure, while with presence of food, all larvae died after 6 days. Chlorfenapyr caused complete mortality of larvae after 5 days of exposure on concrete without food and after 8 days with food. In the case of deltamethrin, 100% mortality was recorded after 7 days in the absence of food and 8 days in the presence of food. Regarding pyriproxyfen, complete mortality was not recorded when food was present, reaching 94.1% 14 days postexposure. However, after 12 days, all larvae died in treatments without food. Although egg hatching was higher in the case of s-methoprene on concrete without food, larval mortality was 100% after 8 days of exposure. Nevertheless, when there was food, 87.3% of the exposed larvae died after 13 days. Therefore, it becomes evident that sanitation of storage facilities before the application of contact insecticides is a key factor for the successful control of T. granarium in the egg stage.

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We investigated the efficacy of two types of modified atmospheres (MA) against adults of the khapra beetle, Trogoderma granarium Everts, and the maize weevil, Sitophilus zeamais Motschulsky, under laboratory conditions. Adults of the above species on wheat were exposed to a carbon dioxide (CO2) concentration of 70% or a low oxygen (O2) concentration of 0.1% for durations of 0.67 (16 h), 1, 2, 4, and 6 d and stored in an environmental chamber set at 28 ± 2°C, 70 ± 5% RH, and a photoperiod of 16:8 (L:D) h. After each exposure interval, immediate mortality and knockdown were recorded, and the surviving or knocked down individuals were transferred to normal atmospheres and returned to the environmental chamber, where survival was recorded 7 d later. In addition, after the immediate and delayed mortality counts, all adults were removed from the substrate, and the number of progeny produced was recorded 60 d later. Both MA conditions totally controlled (100% mortality) the adults of T. granarium and S. zeamais immediately after a 6-d exposure or after a 4-d exposure when delayed mortality was taken into account, showing the postexposure effect of the MA. Moreover, high CO2 was more effective than low O2 for S. zeamais, whereas the reverse was true for T. granarium. The 4-d exposure period was crucial for the production of progeny of both species because after that period surviving insects did not produce offspring. Our results show that both MA conditions can be used with success to control these species.

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Khapra beetle, Trogoderma granarium Everts, is one of the world's most important pests of stored grain. Common in Africa and Asia, it is a quarantine insect for much of the rest of the world where methyl bromide has traditionally been used for its control. However, this ozone-depleting fumigant is now heavily restricted, and alternate methods of control are required. In a two-step process, we examined the use of high-temperature exposure as one such method of control. First, different life stages were held at 45°C for different periods to calculate LT50 (lethal time to 50% mortality) values. In descending order, the most heat-tolerant life stages at 45°C were diapausing larvae (LT50 = 41 - 122 h) > nondiapausing larvae (LT50 = 47 h) > adults (LT50 = 33 h) > pupae (LT50 = 25 h) > eggs (LT50 = 10 h). Second, diapausing larvae (the most heat-tolerant stage) were held at 45, 50, 55, and 60°C for different periods to calculate LT50, LT95, LT99, and probit 9 (99.9968% mortality) values. Estimated LT99 values for diapausing larvae were 288 h at 45°C, 6 h at 50°C, 1.1 h at 55°C, and 1 h at 60°C. Based on these results, an exposure of 2 h at 60°C is recommended to control T. granarium with high temperatures. To meet requirements for control of quarantine pests, exposure of between 2 and 12 h at 50-60°C is recommended to cause probit 9 mortality, but additional experiments are needed to get a better estimate of probit 9.

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In this study, cold hardiness and some physiological characteristics of the Khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae) larvae, were investigated under different thermal regimes, i.e., control, cold-acclimated (CA), fluctuating-acclimated (FA), and rapid cold-hardened (RCH). In all the regimes, the larval survival rate decreased with a decrease in temperature. CA larvae showed the highest cold hardiness following 24 h exposure at -15 and -20°C. Control larvae had the highest glycogen content (34.4 ± 2.3 µg/dry weight). In contrast, CA larvae had the lowest glycogen content (23.0 ± 1.6 µg/dry weight). Change in trehalose content was reversely proportional to changes in glycogen content. The highest myo-inositol and glucose contents were detected in CA larvae (10.7 ± 0.4 µg/dry weight) and control (0.49 ± 0.03 µg/dry weight), respectively. In control and treated larvae, [Na+] decreased, though [K+] increased, with increasing exposure time. The shape of the thermal reaction curve of AMP-depended protein kinase and protein phosphatase 2C followed the same norm, which was different from protein phosphatase 1 and protein phosphatase 2A. Protein phosphatase 2A and 2C showed a complete difference in thermal reaction norms. Indeed, thermal fluctuation caused the highest changes in the activity of the enzymes, whereas the RCH showed the lowest changes in the activity of the enzymes. Our results showed a significant enhancement of larval cold tolerance under CA regime, which is related to the high levels of low molecular weight carbohydrates under this regime. Our results showed that among the different thermal regimes tested, the CA larvae had the lowest supercooling point (about -22°C) and the highest cold hardiness following 24 h exposure at -15 and -20°C.

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Trogoderma granarium Everts (Coleoptera: Dermestidae) is an important insect pest of stored products. In this study, the survival strategies of T. granarium fourth instar larvae were investigated at different sub-zero temperatures following different cooling rates, acclimation to different relative humidity (RH) and different starvation times. Our results show that larvae of T. granarium are freeze-intolerant. There was a strong link between cooling rates and supercooling point, which means the slower the decrease in temperature, the lower the supercooling point. Trehalose content was greater in insects cooled at a rate of 0.5°C/min. According to results, the RH did not affect supercooling point. However, acclimation to an RH of 25% increased mortality following exposure to - 10°C/24h. The time necessary to reach 95% mortality was 1737h and 428h at - 5°C and - 10°C. The lowest lipid and trehalose content was detected in insects acclimated to 25% RH, although, the different RH treatments did not significantly affect glycogen content of T. granarium larvae. The supercooling point of larvae was gradually increased following starvation. By contrast, fed larvae had the greatest lipid, glycogen, and trehalose content, and insects starved for eight days had the lowest energy contents. There was a sharp decline in the survival of larvae between - 11 and - 18°C after 1h exposure. Our results indicate the effects of cooling rate and starvation on energy reserves and survival of T. granarium. We conclude that T. granarium may not survive under similar stress conditions of the stored products.

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