Knockdown of the Expression of Two Trehalase Genes with RNAi Disrupts the Trehalose and Chitin Metabolism Pathways in the Oriental Armyworm, Mythimna separata.

Trehalose is an important carbohydrate substance in insect hemolymph. Chitin is the main component of cuticle and peritrophic matrix in insects. Trehalase (Tre) catalyzes the decomposition of trehalose. Few studies of trehalase in lepidopteran insects have been conducted. Here, the functions of soluble Tre (Tre1) and membrane-bound Tre (Tre2) in the growth and development of Mythimna separata were investigated. We cloned and identified Tre1 and Tre2 cDNA sequences in M. separata. Analysis expression revealed that MsTre1 and MsTre2 were highly expressed in midgut and integument, respectively. The expression of MsTre1 and MsTre2 was highest in the pupal stage. We used RNA interference (RNAi) to inhibit Tre expression in M. separata larvae. Injection of dsMsTre1 or dsMsTre2 resulted in abnormal phenotypes and impeded normal molting. Silencing of MsTre1 and MsTre2 resulted in significant changes in the expression of genes in the trehalose and chitin metabolism pathways, significantly increased the trehalose and glycogen content, and significantly decreased MsTre1 and MsTre2 activity, the glucose content, and the chitin content in midgut and integument. Silencing of MsTre1 slowed larval molting, and the new cuticle was significantly thinner. These results indicate that RNAi of Tre may be useful for control strategies against M. separata.

Life-table parameters, functional response, flight ability, and cross-generational effects of matrine demonstrate its safety to Hippodamia variegata (Coleoptera: Coccinellidae).

In Xinjiang's cotton growing area of China, previous studies have shown that matrine is a selective botanical insecticide, with high toxicity to Aphis gossypii Glover (Hemiptera: Aphididae) and low toxicity to its dominant natural enemy, Hippodamia variegata Goeze (Coleoptera: Coccinellidae). However, lethal effects alone are not sufficient evidence to justify introducing matrine into local IPM strategies. In this context, we systematically evaluated the safety of matrine to H. variegata by investigating the effects of contact and stomach toxicity of matrine on the lady beetle's life-table parameters, predatory ability, flight ability of parental adults, and cross-generational effects on life-table parameters of the predator's offspring. We found that matrine at 2,000 mg/l did not have any significant negative effects to adult fecundity, longevity, or the predatory capacity of parental adults of H. variegata. Moreover, it is the same for cross-generational effects of matrine on H. variegate. The contact toxicity of matrine significantly reduced the flight time of H. variegata males, but did not significantly affect flight time and average velocity. Our results support the view that matrine is safe to H. variegata and can be recommended for use in the local IPM strategy for control of A. gossipii.

Lethal and sublethal effects of chlorantraniliprole on the migratory moths Agrotis ipsilon and A. segetum: New perspectives for pest management strategies.

BACKGROUND: Agrotis ipsilon and A. segetum are major migratory pests of many crops in China, and frequent regional outbreaks cause severe yield losses. Use of food attractants is one of the most promising control methods against adult lepidoptera, notably through the attract-and-kill strategy. Chlorantraniliprole's acute toxicity and sublethal effects on both moths were evaluated. RESULTS: Chlorantraniliprole showed high activity against both adults of both species, with LC20 and LC50 values of 0.08 and 0.21 mg L-1 (A. ipsilon), and 0.14 and 0.51 mg L-1 (A. segetum). The fecundity, effective oviposition rate, and egg hatching rate of both species in dual-sex exposure treatments were all significantly reduced compared with the control, and the population growth coefficients in the LC50 ♀ × LC50 ♂ treatments were only 0.32% (A. ipsilon) and 3.35% (A. segetum) that of the control. Furthermore, the flight distance was significantly suppressed from 6.67 km (control) to 0.01 km (LC50 ) for A. ipsilon, and from 7.39 km (control) to 0.78 km (LC50 ) for A. segetum. The proportions of robust- and medium-flight individuals of A. ipsilon and A. segetum in exposure treatments were greatly reduced. CONCLUSIONS: Low lethal concentration exposures to chlorantraniliprole can drastically reduce the reproduction and flight performance of A. ipsilon and A. segetum, while inhibiting the production of offspring, suggesting chlorantraniliprole would be an excellent compound for use in combination with food attractants. Chlorantraniliprole has good potential for management of the two long-range migratory pests tested using an attract-and-kill strategy. © 2022 Society of Chemical Industry.

Transgenerational Effects of a Neonicotinoid and a Novel Sulfoximine Insecticide on the Harlequin Ladybird.

The harlequin ladybird, Harmonia axyridis Pallas (Coleoptera: Coccinellidae), is a generalist predator and an effective biocontrol agent of various insect pests that has been exploited for the control of aphid pests in the greenhouse and field. However, insecticides are widely used to control aphid pests worldwide and the potential non-target effects of sulfoxaflor and imidacloprid for controlling aphid pests towards this biocontrol agent are little known. Although both sulfoxaflor and imidacloprid act on nicotinic acetylcholine receptors of insects, sulfoxaflor has a novel chemical structure compared with neonicotinoids. We assessed the lethal, sublethal and transgenerational effects of sulfoxaflor and imidacloprid on H. axyridis simultaneously exposed via ingestion of contaminated prey and via residual contact on the host plant at LC20 and LC50 doses estimated for the cotton aphid. Imidacloprid significantly reduced the survival of H. axyridis adults compared to sulfoxaflor at the same lethal concentration against cotton aphid. Both concentrations of imidacloprid and sulfoxaflor reduced the proportion of ovipositing females, and both concentrations of imidacloprid and sulfoxaflor, except LC20 dose of sulfoxaflor, reduced the fecundity and fertility of the parental generation. In the progeny of imidacloprid- and sulfoxaflor-exposed parents, both tested LC50 concentrations significantly decreased the juvenile survival rate, and both concentrations of imidacloprid and sulfoxaflor, except LC20 dose of sulfoxaflor, prolonged the development time. Our findings provide evidence of the negative influence of imidacloprid and sulfoxaflor at low lethal concentrations on the harlequin ladybird and on the progeny of exposed individuals, i.e., transgenerational effects. Hence, these findings stress the importance of optimizing the applications of imidacloprid and sulfoxaflor for the control of aphid pests, aiming at preserving the biocontrol services provided by H. axyridis throughout the integrated pest management approach.

Can contamination by major systemic insecticides affect the voracity of the harlequin ladybird?

Systemic neurotoxic insecticides are widely used to control aphid pests worldwide and their potential non-target effects on aphid predators are often unknown. Behavioral responses linked to biological control services are crucial when assessing the compatibility of chemicals with biocontrol organisms. This is particularly relevant for insecticides at low and sublethal concentrations. We studied the acute toxicity and the sublethal effect on the voracity of the generalist predator Harmonia axyridis (Coleoptera: Coccinellidae) caused by the exposure to three systemic insecticides routinely used against aphids. The tested insecticide concentrations were the Lethal Concentration 50% (LC50), 20% (LC20) and 1% (LC1) estimated for the target pest Aphis gossypii (Hemiptera: Aphididae) in a companion study. The survival and the voracity differed among the tested chemicals and concentrations, but only thiamethoxam at LC50 caused a significant predator mortality, and individuals that survived showed a reduced predation rate. The predators showed a density independent functional response after the exposure to most of the insecticide-concentration combinations, while an inverse density dependence of the prey consumption rate was observed for coccinellids exposed to sulfoxaflor and thiamethoxam at their lowest tested concentration. The estimated parameters, i.e., the attack rate and the prey handling time, were affected at higher concentrations by both imidacloprid and sulfoxaflor. These findings stress the importance of carefully evaluating side effects of insecticides at very low concentrations on beneficial arthropods in the risk assessment schemes for sustainable pest control programmes.

Real-time PCR array chip with capillary-driven sample loading and reactor sealing for point-of-care applications.

A major challenge for the lab-on-a-chip (LOC) community is to develop point-of-care diagnostic chips that do not use instruments. Such instruments include pumping or liquid handling devices for distribution of patient's nucleic-acid test sample among an array of reactors and microvalves or mechanical parts to seal these reactors. In this paper, we report the development of a primer pair pre-loaded PCR array chip, in which the loading of the PCR mixture into an array of reactors and subsequent sealing of the reactors were realized by a novel capillary-based microfluidics with a manual two-step pipetting operations. The chip is capable of performing simultaneous (parallel) analyses of multiple gene targets and its performance was tested by amplifying twelve different gene targets against cDNA template from human hepatocellular carcinoma using SYBR Green I fluorescent dye. The versatility and reproducibility of the PCR-array chip are demonstrated by real-time PCR amplification of the BNI-1 fragment of SARS cDNA cloned in a plasmid vector. The reactor-to-reactor diffusion of the pre-loaded primer pairs in the chip is investigated to eliminate the possibility of primer cross-contamination. Key technical issues such as PCR mixture loss in gas-permeable PDMS chip layer and bubble generation due to different PDMS-glass bonding methods are investigated.

Quartz tuning fork biosensor.

The use of quartz tuning forks for biosensor applications is investigated. The basis of the sensor is to coat the tuning fork surfaces with specific biomolecules and measure subsequent mass loading from the selective binding of complementary analytes. Two experimental set-ups are evaluated, direct mechanical excitation and self-excitation. Mechanical excitation is achieved by mounting the fork on a piezoelectric plate and it is found that the change in oscillation amplitude on adsorption can be monitored to give the change in mass. However, a major drawback is that the sensitivity is determined by the Q-factor, which varies significantly between different sensors and different experimental arrangements. In self-excitation mode, tuning fork motion is activated and detected by placing the fork within a tuned circuit. Using self-excitation mode, anti-human IgG modified tuning forks can sense the binding of human IgG in the range of 5-100 microg ml(-1). The significance of this study is that quartz tuning forks are routinely made using standard microfabrication process, thus suggesting the possibility of facile microfabrication of arrays of quartz sensors.