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Bradysia odoriphaga is a major insect pest that infests Chinese chive in northern China. Clothianidin is a second-generation neonicotinoid insecticide that is commonly used against B. odoriphaga. In this study, the effect of sublethal clothianidin concentrations (LC5 and LC10) on key biological characteristics of B. odoriphaga was investigated using an age-stage, two-sex life table method. Bioassays results showed that clothianidin exhibited high toxicity against B. odoriphaga with LC50 of 1.898 mg L−1 following 24 h exposure. The developmental duration of larvae was significantly increased when exposed to the LC5 (0.209 mg L−1) and LC10 (0.340 mg L−1) of clothianidin. No significant effects were observed on the pupal stage, adult pre-oviposition period (APOP), total pre-oviposition period (TPOP), and mean longevities of male and female. The oviposition period and fecundity of B. odoriphaga were reduced in clothianidin-treated groups. Moreover, key demographic parameters, including the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0), were significantly decreased by the LC5 and LC10 of clothianidin, while no effects were noted on mean generation time (T). Overall, this study showed that sublethal concentrations of clothianidin have a detrimental effect on B. odoriphaga developmental period, fecundity, and life table parameters. Therefore, clothianidin has the potential to suppress the population of B. odoriphaga even at sublethal concentrations.

The fruit and shoot borer, Leucinodes orbonalis, is a major pest affecting brinjal production throughout the world. In this study, we investigated the efficacy of various natural plant extracts and a synthetic insecticide against L. orbonalis, and their impact on natural enemies. Eight different treatments were tested as follow: (1) neem oil at 2%, (2) neem oil at 5%, (3) Eucalyptus leaf extracts at 3%, (4) Eucalyptus leaf extracts at 6%, (5) garlic bulb extracts at 4%, (6) garlic bulb extracts at 8%, (7) cypermethrin 25 EC, and (8) untreated control. Results showed the lowest shoot and fruit infestation in cypermethrin-treated plots (2.03%), followed by neem oil (5%). The highest shoot and fruit infestation were recorded in the untreated control, followed by garlic bulb extract (4%). The highest number of flowers per plant was observed in cypermethrin-treated plots, followed by neem oil (5%), while the lowest was recorded in untreated control, followed by garlic bulb extract (4%). The highest population densities of ladybird beetle, green lacewing, and syrphid fly were recorded in the control, while the lowest densities of these natural enemies were observed in cypermethrin-treated plots. The highest yield of brinjal was recorded in cypermethrin-treated plots (3731.5 kg/ha), followed by neem oil at 5% with 3273.2 kg/ha, while the lowest yield was obtained in control plots (753.3 kg/ha). Overall, this study showed that the natural plant extracts (notably neem oil 5%) significantly reduced shoots and fruit infestation, and could, therefore, be used against L. orbonalis in the brinjal crop.

The melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae) is a major pest affecting plants of Cucurbitaceae family worldwide. Clothianidin has been broadly used for the control of this insect pest. In this study, we investigated the impact of clothianidin on the demographical traits and expression changes of vitellogenin and ecdysone receptors genes in A. gossypii after exposure to the low or sublethal concentrations (LC15 and LC5, respectively). The two concentrations of clothianidin did not affect the adult longevity and fecundity of parental aphids (F-0). However, the developmental duration of 3rd instar nymph, pre-adult stage, and total pre-ovipositional period (TPOP) were increased significantly in F-1 individuals when F o was exposed to the LC15 of clothianidin. In contrast, the adult longevity and fecundity of progeny generation were decreased by the LC15. Similarly, the net reproductive rate (R-0) and gross reproduction rate (GRR) were also reduced significantly in the LC15 treated group. However, no significant effects were observed for the intrinsic rate of increase (r), finite rate of increase (lambda) and mean generation time (T). Furthermore, the expression level of vitellogenin receptor gene significantly decreased by 24.9 and 56.9% in the progeny generation when F-0 was exposed with LC5 and LC15 of clothianidin, respectively. Moreover, the ecdysone receptor mRNA expression was also reduced by 29.2 and 46.1% in the LC5 and LC15 treated groups, respectively. These results indicated that clothianidin at low or sublethal concentrations adversely affect the development and reproduction of progeny generations and effects may be related to the down-regulation of VgR and EcR genes in A. gossypii.

Cotesia flavipes Cameron is an important larval parasitoid exploited for the control of the spotted stem borer, Chilo partellus (Swinhoe). Several studies have evaluated the toxic effects of insecticides on C. partellus, however, little is known about non-target effects of insecticides on this parasitoid, when used to control C. partellus. This laboratory study evaluated the lethal and sublethal effects of twelve insecticides on C. flavipes. Residual toxicity tests showed that organophosphates (chlorpyrifos, triazophos and profenofos) exhibited highest contact toxicity to C. flavipes adults with a LC50 range from 0.63 to 1.05 mg a.i/l, while neonicotinoids (nitenpyram, acetamiprid and imidacloprid) were less toxic to C. flavipes with a LC50 range from 1.27 to 139.48 mg a.i/l. Sugar-insecticide feeding bioassays showed that organophosphates, pyrethroids (cypermethrin, bifenthrin and lambda-cyhalothrin) and carbamates (thiodicarb, carbaryl and methomyl) were highly toxic to C. flavipes adults and caused 100% mortality at 48 h of exposure, while imidacloprid caused 66% mortality at 48 h of exposure. Risk quotient analysis showed that among all tested insecticides, imidacloprid and acetamiprid were less toxic to C. flavipes adults with a risk quotient value of 0.88 and 1.6, respectively. Furthermore, exposure of immature C. flavipes through their host bodies significantly decreased the parasitism rate at their F1 and F2 generations. Risk quotient analysis of insecticides indicated that imidacloprid and acetamiprid were the least toxic to C. flavipes. This study provides important information that will be used in incorporating the most suitable insecticides in integrated pest management programs with reduced negative impacts on non-target beneficial arthropods.

Wheat (Triticum aestivum L.) is one of the major source of food worldwide. However, it suffers from numerous constraints. The aphids, particularly Rhopalosiphum padi (Linnaeus) and Sitobion avenae (Fabricius) species are among the major causes that severly affect wheat production acrossthe globe. To determine the population dynamics of aphids associated with wheat crop in Khyber Pakhtunkhwa, we examined the mean population of both species in the selected areas (Maho Dheri, Sarband, Kandare, Rag Narai and Khazana Dheri) of district Mardan in 2015. The mean population of R. padi and S. avenae per plant were recorded every week by counting the number of aphids on 5 plants picked up randomly from each selected wheat field using the diagonal methods. The results on current study showed that the infestation of both aphid species was started in the 1st week of February (05 standard weeks). The densities of aphids per plant increased as the vegetative growth proceeded and was highest till the mid of March (10th and 11th standard week). The declined in aphid populations for both species were started after mid of March and ended in April. The mean population of both aphid species was found low in the 1st week of February (05th standard week) that is probably due to the low temperature. The population density of aphids reached to the its peak in the second week of March due to suitable temperature for aphid infestation. Keywords: District Mardan; Population dynamics; Rhopalosiphum padi; Sitobion avenae and Wheat http://dx.doi.org/10.19045/bspab.2020.90004

Spotted bollworm, Earias vittella (Fab), is an agricultural cotton pest. In Pakistan, Earias vittella (Fab) has developed resistance to various groups of insecticides. In the present study, the effect of an extract of Azadirachtin indica in different solvents on second instar larvae of E. vittella was evaluated in laboratory studies for toxicity as well as lethal and sublethal effect. The results indicated that methanol-based extract was the most effective to control E. vittella, followed by ethanol extract. The lethal and sublethal concentration of methanol-based extract of A. indica increased larval and pupal duration as compared to control. The percent pupation, adult emergence, and pupal weight were decreased in treated populations as compared to control populations. The esterase activities and total protein content of the treated E. vittella were also reduced as compared to controls. The results of this study could be useful in an integrated pest management program for the management of E. vittella.

MicroRNAs (miRNAs) regulate various biological processes during insect development; however, their role in larval-pupal development in oriental fruit fly, Bactrocera dorsalis (Hendel) remains unknown. In the current study, we address the biological function of a conserved miRNA, Bdo-Let-7 in the regulation of BdE75 gene, which belongs to the ecdysone signaling pathway and participates in the larval-pupal development in B. dorsalis. Using dual luciferase reporter assay in HEK293T cells we show that Bdo-Let-7 miRNA interacts with the 3' untranslated region of BdE75 gene and suppresses its expression. The Bdo-Let-7 and BdE75 are also co-expressed in the larval-pupal stages and in different tissues of B. dorsalis. In in vivo experiments, the injection of Bdo-Let-7 agomir and antagomir in third instar larvae down- and up-regulated the expression of BdE75, respectively. The 20-hydroxyecdysone (20E) injection assay shows that 20E up-regulated the expression of Bdo-Let-7 on the 5th day of the larvae. Moreover, abnormal pupation and eclosion were observed after larval Bdo-Let-7 antagomir injection. Based on these results, we show that Bdo-Let-7 regulates the ecdysone signaling pathway through the exact dose of BdE75 gene, and is indispensable for normal larval-pupal development in B. dorsalis.

Detrimental effects of ultraviolet (UV) light on living organisms are well understood, little is known about the effects of blue light irradiation. Although a recent study revealed that blue light caused more harmful effects on insects than UV light and blue light irradiation killed insect pests of various orders including Diptera, the effects of blue light on physiology of insects are still largely unknown. Here we studied the effects of blue light irradiation on cuticular melanin in larval and the immune response in adult stage of Bactrocera dorsalis. We also evaluated the effects of blue light exposure in larval stage on various age and mass at metamorphosis and the mediatory role of cuticular melanin in carryover effects of larval stressors across metamorphosis. We found that larvae exposed to blue light decreased melanin contents in their exoskeleton with smaller mass and delayed metamorphosis than insects reared without blue light exposure. Across metamorphosis, lower melanotic encapsulation response and higher susceptibility to Beauveria bassiana was detected in adults that had been exposed to blue light at their larval stage, thereby constituting the first evidence that blue light impaired adult immune function in B. dorsalis as a carryover effect of larval exposure.

The genetic-based sterile insect technique (SIT) is an effective and environmentally safe strategy to diminish populations of agricultural and horticultural insect pests. Functional characterization of genes related to male fertility can enhance the genetic-based SIT. Tssk1 has been involved to control male fertility in both mammals and insects. Moreover, Tektin1 has also been revealed to influence male fertility in both human and mammals. These findings suggested that Tssk1 and Tektin1 identified from Bactrocera dorsalis could be required for male fertility in B. dorsalis. In this study, expression profiles of these two genes were studied at different developmental stages and in various tissues of adult males. Remarkably, it was found that Tssk1 and Tektin1 were highly expressed in the testis of mature adult males of B. dorsalis. Furthermore, Tssk1 and Tektin1 genes were downregulated by using the RNA interference (RNAi) method. Fertility assays including egg laying, hatching, and spermatozoa count were also performed to investigate male fertility of B. dorsalis. Results showed that knockdown of Tssk1 and Tektin1 caused male sterility up to 58.99% and 64.49%, respectively. As expected, the total numbers of spermatozoa were also significantly reduced by 65.83% and 73.9%, respectively. These results suggested that male sterility was happened wing to the low number of spermatozoa. In conclusion, we demonstrate that Tssk1 and Tektin1 are the novel agents that could be used to enhance the genetic-based SIT, or their double-stranded RNA (dsRNA) can be used as biopesticides to control the population of B. dorsalis.

Voltage-gated sodium channels (VGSC) are transmembrane proteins that generate an action potential in excitable cells and play an essential role in neuronal signaling. Since VGSCs play a crucial role in nerve transmission they have become primary targets for a broad range of commercial insecticides. RNA interference (RNAi) is a valuable reverse genetics tool used in functional genomics, but recently, it has also shown promise as a novel agent that could be used to control agricultural insect pests. In this study, we targeted the VGSC (MpNav) gene in the peach-potato aphid Myzus persicae, by oral feeding of artificial diets mixed with dsRNAs. Knock-down of MpNav gene expression caused up to 65% mortality in 3rd instar nymphs. Moreover, significantly lower fecundity and longevity was observed in adult aphids that had been fed with dsMpNav solution at the nymphal stage. Analysis of gene expression by qRT-PCR indicated that the aphid mortality rates and the lowered fecundity and longevity were attributable to the down-regulation of MpNav by RNAi. Taken together, our results show that MpNav is a viable candidate target gene for the development of an RNAi-based bio-aphicide.

The melon aphid, Aphis gossypii is a globally distributed crop pest with a wide host range. The intensive use of insecticides against this insect over several years has led to develop resistance against many insecticides including acetamiprid. Understanding the relationship between acetamiprid resistance and fitness of A. gossypii is essential to limit the spread of the resistant population in the field. In this study, age-stage, two-sex life table approach was used to investigate these relationships in the lab. Results showed that resistant strain (Ace-R) had a reduced fitness (relative fitness = 0.909) along with significantly decreased adult longevity, fecundity, net reproductive (R0), mean generation time (T) and gross reproductive rate (GRR). Compared to the susceptible strain (Ace-S), the pre-adult period and total pre-oviposition period (TPOP) were also significantly shorter in Ace-R strain. Moreover, the expression profiles of EcR, JHBP, JHAMT, JHEH, USP and Vg genes supposed to be involved in insect reproduction and development were analyzed using Quantitative Real Time PCR. The EcR, JHBP, JHAMT and USP genes were up-regulated, Vg gene was down-regulated while the mRNA level of JHEH gene was statistically same in the Ace-R strain compared to the Ace-S strain. Collectively, this study provides the occurrence and magnitude of fitness costs of A. gossypii against acetamiprid resistance and could be helpful to manage the resistance evolution in field populations.

International audience; Clothianidin is a second-generation neonicotinoid insecticide, widely used against sap-sucking insect pest including melon aphid,Aphis gossypiiGlover (Hemiptera: Aphididae). This pest causes severe economic damage to Cucurbitaceae plants worldwide. In this study, we investigated clothianidin resistance development under continuous selection pressure. Moreover, the age-stage, two-sex life table approach was used to evaluate the impact of clothianidin resistance on the fitness ofA.gossypii. A clothianidin resistant strain (CT-R) with a 23.17-fold resistance level was developed from a susceptible strain (CT-S) after continuous selection for 24 generations. Life table results showed a significant reduction in the relative fitness (0.847) of CT-R strain compared to the CT-S strain ofA.gossypii. The developmental duration, oviposition days, total pre-oviposition period (TPOP), longevity, and fecundity of CT-R strain were found to be significantly lower when compared to CT-S strain. The demographic parameters, including the intrinsic rate of increase (r), finite rate of increase (lambda), net reproductive rate (R-0), and mean generation time (T) were also significantly decreased in CT-R strain compared to the CT-S strain. Both the reproductive and survival rates were affected by clothianidin resistance in CT-R strain compared with the CT-S strain ofA.gossypii. Overall, our results demonstrate that in-depth knowledge about the trade-off at play between resistance degree and fitness cost might be useful to design resistance management strategies againstA.gossypii.

Insecticide induced-hormesis, a bi-phasic phenomenon characterized by low dose stimulation and high dose inhibition following exposure to insecticide, is crucial to insect pest resurgence. In this study, the effects of low or sublethal concentrations of thiamethoxam on biological traits and genes expression were investigated for Aphis gossypii Glover following 72 h exposures. Leaf-Dip bioassay results showed that thiamethoxam was very toxic against adult A. gossypii with an LC50 of 1.175 mg L−1. The low lethal (LC15) and sublethal (LC5) concentrations of thiamethoxam significantly reduced longevity and fecundity of the directly exposed aphids. However, stimulatory effects on pre-adult stage, longevity, and fertility were observed in the progeny generation (F1) of A. gossypii, when parental aphids (F0) were exposed to LC15 of thiamethoxam. Subsequently, biological traits such as intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0) increased significantly to F1 individuals due to LC15 treatment. No significant responses were observed for LC5 of thiamethoxam. The LC15 of thiamethoxam significantly increased the expression level of vitellogenin and ecdysone receptors genes in progeny generation, while no effects were observed for treatment with LC5. Additionally, the expression levels of P450 genes including CYP6CY14, CYP6CZ1, CYP6DC1, CYP6CY9, and CYP6DD1 were up-regulated in the exposed aphids. Taken together, our results show the hormetic effects of thiamethoxam on F1 individuals, which might be due to the intermittent changes in expression of genes involved in fertility, growth and insecticide detoxification in A. gossypii.

Voltage-gated sodium channels (VGSC) are transmembrane proteins that generate an action potential in excitable cells and play an essential role in neuronal signaling. Since VGSCs play a crucial role in nerve transmission they have become primary targets for a broad range of commercial insecticides. RNA interference (RNAi) is a valuable reverse genetics tool used in functional genomics, but recently, it has also shown promise as a novel agent that could be used to control agricultural insect pests. In this study, we targeted the VGSC (MpNav) gene in the peach-potato aphid Myzus persicae, by oral feeding of artificial diets mixed with dsRNAs. Knock-down of MpNav gene expression caused up to 65% mortality in 3rd instar nymphs. Moreover, significantly lower fecundity and longevity was observed in adult aphids that had been fed with dsMpNav solution at the nymphal stage. Analysis of gene expression by qRT-PCR indicated that the aphid mortality rates and the lowered fecundity and longevity were attributable to the down-regulation of MpNav by RNAi. Taken together, our results show that MpNav is a viable candidate target gene for the development of an RNAi-based bio-aphicide.

Phytoseiid mites are considered the most effective natural enemies of pest mites. They also have been shown to attack pest thrips. It is unknown, however, whether phytoseiid mites can reduce high densities of Thrips flavidulus (Bagnall). We addressed this question by the study of functional and numerical responses. The aim of this research was to evaluate the potential predation success of the adults of three predatory mites, Neoseiulus cucumeris (Oudemans), Neoseiulus barkeri (Hughes), and Euseius nicholsi (Ehara & Lee), against the first-instar of T. flavidulus in a climatic chamber at five different temperatures. The results showed that the functional responses of those predators reflected the Holling type II functional response and were density dependent and positively related to temperature. For the three predatory mites, predation and successful attack rates increased with increasing temperature up to 26°C, reducing afterward. Handling time had the opposite trend. Reproductive ability also increased with an increase in temperature and prey consumption.

The cotton leafhopper, Amrasca biguttula, is one of the most destructive pests of cotton in Asia. This species is thought to cause damage by injecting enzymatic saliva into various, presently unknown, cotton tissues and ingesting the resulting macerate. Flonicamid is a novel systemic insecticide used to control the cotton leafhopper; however, its mode of action is unknown.The mechanism of action of flonicamid on cotton leafhopper was investigated using electropenetrography (EPG). EPG recordings revealed six waveforms, i.e. Np (non-probing), A1 (channel cutting), A2, A3, A4, A5 and A6. Waveforms A2 and A3 probably represent active ingestion with (A2) and without (A3) simultaneous watery salivation. The meanings of A4, A5 and A6 are presently unknown, but minor in duration. Flonicamid significantly increased the mean duration of non-probing events and strongly inhibited ingestion by treated insects, which resulted in the slow death of leafhoppers. Inhibition of ingestion was dose dependent, and near-complete suppression was observed when the flonicamid concentration was increased to 10 000 mg LWe propose that starvation caused by inhibition of active ingestion is the mechanism of toxicity for flonicamid. This knowledge could aid in applicability and use of this new insecticide for field management of leafhopper populations. © 2016 Society of Chemical Industry.

Genetics-enhanced sterile insect techniques (SIT) are promising novel approaches to control Bactrocera dorsalis, the most destructive horticultural pest in East Asia and the Pacific region. To identify novel genetic agents to alter male fertility of B. dorsalis, previous studies investigated miRNA expression in testes of B. dorsalis. One miRNA, miR-8-3p was predicted to bind the 3′UTR of putative B. dorsalis mitoferrin (bmfrn). The ortholog of bmfrn in D. melanogaster is essential for male fertility. Here we show that bmfrn has all conserved amino acid residues of known mitoferrins and is most abundantly expressed in B. dorsalis testes, making miR-8-3p and mitoferrin candidates for genetics-enhanced SIT. Furthermore, using a dual-luciferase reporter system, we show in HeLa cells that miR-8-3p interacts with the 3′UTR of bmfrn. Dietary treatments of adult male flies with miR-8-3p mimic, antagomiR, or bmfrn dsRNA, altered mitoferrin expression in the testes and resulted in reduced male reproductive capacity due to reduced numbers and viability of spermatozoa. We show for the first time that a mitoferrin is regulated by a miRNA and we demonstrate miR-8-3p as well as bmfrn dsRNA to be promising novel agents that could be used for genetics-enhanced SIT.

MicroRNAs (miRNAs) are a class of endogenous small non-coding RNAs that regulate various biological processes including sexual dimorphism. The oriental fruit fly Bactrocera dorsalis is one of the most destructive agricultural insect pests in many Asian countries. However, no miRNAs have been identified from the separate sex and gonads to elucidate sex gonad differentiation in B. dorsalis. In this study, we constructed four small RNA libraries from whole body of females, males (except ovaries and testes) and ovaries, testes of B. dorsalis for deep sequencing. The data analysis revealed 183 known and 120 novel miRNAs from these libraries. 18 female-biased and 16 male-biased miRNAs that may be involved in sexual differentiation were found by comparing the miRNA expression profiles in the four libraries. Using a bioinformatic approach, we predicted doublesex (dsx) as a target gene of the female-biased miR-989-3p which is considered as the key switch gene in the sex determination of tephritid insects. This study reveals the first miRNA profile related to the sex differentiation and gives a first insight into sex differences in miRNA expression of B. dorsalis which could facilitate studies of the reproductive organ specific roles of miRNAs.

Ultraviolet (UV) light has been used worldwide to monitor and trap insect pests. Whitefly adults show conspicuous positive phototactic behavior toward UV light stimuli; however, knowledge of the effect of UV light exposure on various life-history parameters of Dialeurodes citri remains limited. The present research aimed to investigate the effect of ultraviolet radiation (UV-A; long-wave) exposure on the reproduction and longevity of D. citri adults as well as the development of immature (eggs, larvae, and pupae) flies in the F1 generation. Paired D. citri adults were exposed to UV-A radiation for different periods (0, 1, 4, and 7 h/d) until the end of their life. The results of the experiment revealed that fecundity and oviposition rates increased when adults were irradiated for 1 and 4 h/d, but interestingly, both were significantly decreased compared with those of the controls after the longest exposure time (7 h/d). The longevity of adults of both sexes and the cumulative survival of F1 immatures were decreased with increased exposure time. Exposure to UV-A radiation prolonged the developmental time of immature stages, and a positive correlation was observed with exposure time. Exposure to UV light significantly inhibited egg hatching, larval development, pupation, and adult emergence. To the best of our knowledge, this is the first study describing the effect of UV radiation on a homopteran insect pest. This research may provide a foundation for the scientific community to use UV light in the field as an integrated pest management strategy to control this devastating agricultural pest.