Exposure to herbicides reduces larval sensitivity to insecticides in Spodoptera litura (Lepidoptera: Noctuidae)

Herbicides and insecticides are widely used in modern agriculture. It has been reported in various studies that application of insecticides can increase tolerance of herbivorous insects to insecticides. However, limited information exists on susceptibility to insecticides when insects are exposed to herbicides. This study was conducted to investigate the potential impact of the herbicides trifluralin and 2-methyl-4-chlorophenoxyacetic acid sodium salt (MCPA-Na) on the susceptibility of the nocturnal moth Spodoptera litura to the insecticides X-cyhalothrin, phoxim and bifenthrin. We found that larvae exposed to trifluralin or MCPA-Na became significantly less susceptible to both insecticides than nonexposed control larvae. Herbicide-treated larvae did not show altered growth under the used test conditions. However, heads of herbicide-treated larvae showed increased expression of the acetylcholinesterase genes SI Ace I and SI Ace 2. Moreover, the fat body and midgut of herbicide-treated larvae displayed elevated expression of detoxification genes (the carboxylesterase gene SI CarE;the glutathione S-transferase genes SlGSTe2 and SlGSTe3\ the cytochrome P450 monooxygenase genes CYP6B48, CYP9A40 and CYP321B1). The CYP6B48 gene exhibited highest inducibility. In conclusion, the data of this study suggest that exposure of S. litura larvae to herbicides may stimulate detoxification mechanisms that compromise the efficacy of insecticides.     

Candidate insecticides for the control of larvae of Mamestra brassicae (Lepidoptera) (Noctuidae)

Fifteen insecticides were tested in the laboratory against larvae of Mamestra brassicae, a chrysanthemum pest. Seven of these, methomyl, chlorpyrifos, leptophos, iodofenphos, methamidophos, mephosfolan and fenitrothion were significantly more potent than carbaryl, the standard adopted. The performance of the insecticides is discussed in relation to selective action against insects and to mammalian toxicity, to identify those which might be suitable for integrated control or chemical control of noctuid larvae under glass. A technique for monitoring the amount of pesticide deposited on to a target by a Potter Tower sprayer is described.     

Sensitivity of polyphagous (Plodia interpunctella) and stenophagous (Ephestia kuehniella) storage moths to residual insecticides: effect of formulation and larval age

Pyralid moths, Ephestia kuehniella and Plodia interpunctella, are prevalent stored product pests. The insecticides are the main tool to control these moths in the stores. The data describing the response of these moths to insecticides are scarce. The lethal effect of the organophosphate, pyrethroid, and halogenated-pyrrole on moths larvae were compared in laboratory test. The hypothesis was that the very polyphagous P. interpunctella would have generally higher insecticide tolerance than that of the stenophagous E. kuehniella. Different insecticide concentrations were applied onto the inner surface of glass tube vials. Ten larvae of 14 or 21 d old of E. kuehniella and 7 or 14 d old of P. interpunctella were used by treatment. The larval mortality was checked after 24 h of exposure. The mortality was significantly influenced by age of larvae and the groups of chemicals. No differences among the efficacies of the tested formulations with identical active compounds were found, except significant different mortality of E. kuehniella on deltamethrin formulations. A comparison of analytical standards showed that P. interpunctella was less susceptible to the active ingredient pirimiphos-methyl than E. kuehniella, while E. kuehniella was less susceptible to deltamethrin than P. interpunctella. No differences between the two species were observed for chlorfenapyr. We therefore rejected the hypothesis that polyphagy/stenophagy can be a general predictor of insecticide tolerance in the two tested storage moths. The most important finding for effective use was that the young larvae of both species were more susceptible to tested insecticides than older larvae.     

Effect of proteolytic and detoxification enzyme inhibitors on Bacillus thuringiensis var. israelensis tolerance in the mosquito Aedes aegypti

Bacillus thuringiensis var. israelensis (Bti) is highly pathogenic to mosquito larvae and is widely used for mosquito control. Its mosquitocidal activity however is relatively low compared to many chemical insecticides. The detoxification mechanisms in the mosquito, among other things, might neutralize the Bti activity, resulting in resistance or tolerance. We tested whether or not the detoxification mechanisms against chemical insecticides might also operate against Bti, rendering it less effective. We targeted four enzymes in Aedes aegypti larvae involved in detoxification with inhibitors that have been used in resistance studies in chemical insecticides and assayed their effects on Bti toxicity. Results revealed that phenylmethanesulphonyl fluoride (PMSF), diethyl maleate, phenobarbital (PB), and piperonyl butoxide (PBO) altered Bti toxicity to various degrees. PMSF is a serine protease inhibitor that prevents Bti digestion and improves Bti activity. PB that induces several detoxifying enzymes had two different effects depending on the method of treatment. Mortality was higher when treatment with PB was discontinuous (149%) whereas with continuous treatment it was lower (101%). PBO, a typical cytochrome P450 inhibitor, increased Bti effect (159%). The combination of discontinuous pretreatment of larvae with PB followed by PBO had a synergistic effect and showed increased activity (146%). It appears that the mechanism for Bti resistance in mosquitoes is similar to that of chemical insecticides. Our studies indicate that we may be able to increase Bti activity by inhibiting some of the detoxification systems as active as broad spectrum chemical insecticides.     

Insect growth regulatory and larvicidal activity of chalcones against Aedes albopictus

Widespread use of chemical insecticides has resulted in development of insect resistance and natural products with biological activities could become an attractive alternative to control insect pests. In order to find more effective insecticides for controlling mosquito, various mosquitocidal compounds are studied. Recently, juvenile hormone antagonists (JHANs) have been found to be to safe and effective insecticides for control of mosquito. In order to identify novel insecticidal compounds with JHAN activity, several chalcones were surveyed on their JHAN activities and larvicidal activities against Aedes albopictus larvae. Among them, 2′‐hydroxychalcone and cardamonin showed high levels of JHAN and mosquito larvicidal activities. These results suggested that chalcones with JHAN activity could be useful for control of mosquito larvae.     

Evaluation of larvicidal potential of certain insect pathogenic fungi extracts against Anopheles stephensi and Culex quinquefasciatus

Fungal metabolites are attracting attention as potential microbial insecticides, and they are anticipated to overcome the problems of pesticide resistance and environmental pollution that are associated with the indiscriminate use of conventional synthetic insecticides. The relative bioefficacies of selected fungal pathogens, Aspergillus flavus, A. niger, A. parasiticus, Fusarium sporotrichoides and Penicillium verrucosum were observed against Anopheles stephensi and Culex quinquefasciatus larvae. A. flavus demonstrated the greatest bioefficacy with 50% lethal concentration (LC₅₀) values of 9.54 and 10.98 ppm against Anopheles stephensi and Culex quinquefasciatus larvae, respectively, after 24‐h exposure. The bioefficacy of A. flavus increased in both species with an exposure time of 48 h, with LC₅₀ values of 7.26 and 8.55 ppm, respectively.     

Synergies between biological and neonicotinoid insecticides for the curative control of the white grubs Amphimallon majale and Popillia japonica

Synergistic combinations of biological and chemical insecticides might yield promising alternatives for soil insect pest management. In turfgrass of the Northeast U.S., control of root-feeding scarab larvae is highly dependent on conventional insecticides. Studies on interactions between entomopathogenic nematodes and neonicotinoid insecticides, however, demonstrate the feasibility of synergies as an approach for reduced-risk curative control. To understand the breadth of potential synergies, we screened numerous combinations of biological control agents with sublethal doses of neonicotinoids against third instars. Interactions were characterized as synergistic, additive or antagonistic. The most promising combinations identified in laboratory bioassays were advanced to greenhouse pot studies and then to field trials featuring microplots with artificially infested populations. To reveal variation across scarab species, trials were conducted on Amphimallon majale and Popillia japonica. Synergies were consistent across trials and specific to white grub species. For A. majale, synergistic combinations of Heterorhabditis bacteriophora with imidacloprid and clothianidin were discernible in laboratory, greenhouse and field trials. For P. japonica, synergistic combinations of Beauveria bassiana and Metarhizium anisopliae with both neonicotinoids were discernible in the laboratory and greenhouse, but not in the field. For both species, antagonistic interactions were discernible between Bt-products and both neonicotinoids. While nematode-neonicotinoid synergies among scarab larvae have been examined before, fungi-neonicotinoid synergies are unreported. In the context of previous studies, however, no patterns emerge to explain variation across target species or control agent. Further study of non-additive interactions will guide how biological and chemical products could be combined to enhance soil insect pest management.     

Formulation and application of the entomopathogenic fungus: Zoophthora radicans (Brefeld) Batko (Zygomycetes: Entomophthorales)

Aims: To isolate and formulate a native strain of Zoophthora radicans naturally infecting larvae of diamondback moth, Plutella xylostella, existing in South Australia and to provide evidence that formulation of the fungus is effective against P. xylostella larvae, and therefore, it could be used as a tool in pest management of this insect. Methods and Results: Dose–response bioassays using formulated and unformulated forms of the fungus strain were carried out against third instar larvae of P. xylostella. Results obtained have indicated a significant increase in the larval mortality when higher concentrations of a formulated form of the fungus strain were applied compared to the treatments with the unformulated form (85·0 vs 57·5% of larval mortality, respectively, at the top concentration of 10⁷ conidia/ml). The median lethal concentration (LC50) for a formulated form was 100 times less than that of the unformulated form when they were applied against the third instar larvae of P. xylostella. In addition, the formulation used in the present bioassays has preserved the viability of introduced fungus conidia for longer time in comparison with the unformulated conidia. Conclusions: The effective application of a formulated fungus strain against P. xylostella larvae constitutes the first step towards its use in pest management of this insect. Significance and Impact of the Study: The formulated fungus in inverted emulsion could be used as an alternative tool to insecticides in pest management of P. xylostella larvae because of the development of resistance to insecticides in the treated larvae.     

Molecular identification of glutathione S-transferase genes and their potential roles in insecticides susceptibility of Grapholita molesta

Grapholita molesta is one kind of serious fruit-boring pests in the world, which caused huge economic losses to the fruit industry including pear and apple. The heavy usage of synthetic insecticides resulted in different degrees of resistance of G. molesta to some commonly used insecticides. However, the tolerance mechanisms of G. molesta to insecticides are still unclear. Glutathione S-transferases (GSTs) belong to the superfamily of detoxifying enzymes and mainly play important roles in detoxification metabolism and insecticide susceptibility of insects. In this study, we first evaluated the toxicity of three insecticides including emamectin benzoate, chlorantraniliprole and lambda-cyhalothrin on G. molesta at different developmental stages. Synergism study showed that the susceptibility of G. molesta larvae to these three insecticides obviously increased when the enzyme activity of GST was inhibited by diethyl maleate compared with the controls. Next, we screened out six GmGST genes which are upregulated after insecticides treatment from a total of 21 GmGST genes and analysed their sequence characteristics and expression profiles. RNAi and bioassay results further revealed that the mortality of 5th instar larvae significantly increased after exposure to LC₃₀ of emamectin benzoate, chlorantraniliprole and lambda-cyhalothrin when silencing of GmGSTo2, GmGSTs1 and GmGSTz1 compared with the controls. In conclusion, this study indicated that GmGSTo2, GmGSTs1 and GmGSTz1 play important roles in insecticides susceptibility of G. molesta to three commonly used insecticides, which is significance for its future integrated management.     

Larvicidal potency of selected xerophytic plant extracts on Culex pipiens (Diptera: Culicidae)

Chemical insecticides released into the environment may have adverse biological effects. Therefore, there is a need for ecofriendly insecticides for mosquito control. Xerophytic plant extracts that may provide more ecofriendly active component were evaluated against Culex pipiens 4th instars. Plant extracts prepared using different solvents with a Soxhlet apparatus and different concentrations were tested against Culex pipiens larvae. The effects were observed at 24 h and 72 h intervals and LD₅₀ and LD₉₀ values determined. Chloroform (CHCl₃) and ethyl acetate (EtOAc) extracts of Althaea ludwigii were the most effective against Cx. pipiens 4^th instars, but were highly dependent on extract concentrations and exposure time. Results suggest that A. ludwigii extracts contain bioactive compounds, such as phenols and saponins, that may provide effective Cx. pipiens larval control. However, the extract was found to be toxic to zebrafish larvae, and may be toxic to other aquatic fauna. Further studies to determine the active components and toxicity to other fauna are needed.     

Comparative study of insecticide susceptibility and activities of detoxification enzymes in larvae and adults of cotton bollworm,Heliothis armigera

Susceptibility of moths and larvae of cotton bollworm to ten different insecticides by topical application and their effect on enzymes involved in insecticide detoxification were determined. The moths were more susceptible than larvae to the insecticides tested, with the exception of pyrethroids and sulprofos. Combination of several insecticides with the synergists piperonyl butoxide (PB) and S,S,S-tributylphosphorotrithioate (TBPT) showed that lower level of carbamate and organophosphate toxicity in larvae, as compared to moths, was the result of higher detoxification enzyme activities. Studies of the post-treatment fate of ¹⁴C-labeled malathion and ³H-trans-permethrin indicated that both the cuticular penetration, internal accumulation, and excretion of applied toxicants and their metabolites occurred more rapidly in larvae than in moths. The activities in vitro of esterases, glutathione S-transferases, and monooxygenases were determined but there were no correlations with either toxicity of insecticides or synergistic effect for combination of insecticides with PB and TBPT in moths. © 1996 Wiley-Liss, Inc.     

5种杀虫剂对黑粪蚊的毒力及对平菇菌丝生长的影响

[目的]黑粪蚊为食用菌生产中的危险害虫之一,通过杀虫剂的毒力测定和对平菇菌丝生长抑制试验,拟筛选获得理想的黑粪蚊防治杀虫剂。[方法]采用毒饵法测定了5种杀虫剂对黑粪蚊幼虫的室内毒力,并研究各杀虫剂对平菇菌菌丝生长的抑制情况。[结果]20%甲氰菊酯乳油、14%阿维·虫螨腈悬浮剂和20%甲维·吡丙醚悬浮剂对黑粪蚊的校正死亡率达80%以上,对黑粪蚊幼虫均有较好的杀灭效果,而苏云金杆菌（以色列亚种）悬浮剂对黑粪蚊幼虫的杀虫效果不理想。5种杀虫剂对平菇菌丝生长均有一定抑制作用,且农药品种间存在极显著差异,20%甲氰菊酯乳油抑制率最高,高浓度480 mg·L^-1处理的抑制率可达16.85%,14%阿维·虫螨腈悬浮剂的抑制率最低,高浓度140 mg·L^-1处理的抑制率仅为5.83%。[结论]本研究中14%阿维·虫螨腈悬浮剂用于平菇菌中黑粪蚊防治效果最好,研究结果将为食用菌黑粪蚊的药剂防治提供理论指导。     

Laboratory pesticide screening method for the aphid predatory midge Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae)

A laboratory pesticide screening method is described for the aphid predatory midge Aphidoletes aphidimyza. Four acaricides, 10 fungicides, 11 insecticides and one herbicide, all of which are in common usage, were evaluated against both the adult and larval stages of the predator. The acaricide dienochlor, the fungicides benomyl, bupirimate, chlorothalonil, iprodione and hexaconazole, the herbicide tralkoxydim and the insecticides Bacillus thuringiensis and diflubenzuron were all safe to both adult and larval A. aphidimyza. Adults were more susceptible to the pesticides than larvae. Pesticide was exposed to adults as a residue on glass plates and to larvae on a leaf section mounted on agar. The leaf material remained healthy and supported host aphids for several days without deteriorating. The method and apparatus have potential for use with many other pests and beneficial organisms required to be caged for several days on detached leaf material.     

Toxicity of insecticides to obliquebanded leafroller, Choristoneura rosaceana, larvae and adults exposed previously to neem seed oil

Adult and sixth instar obliquebanded leafroller, Choristoneura rosaceana (Harris), exposed previously as fifth instar larvae to sub-lethal concentrations of neem, Azadirachta indica A. Juss., seed oil contained in artificial diet were more susceptible to topically applied pyrethroid, carbamate and organophosphate insecticides than C. rosaceana reared on control diets. Increased susceptibility to insecticides did not result from reduced vigour, as measured by reductions in adult or larval weights, but is instead correlated with previously demonstrated reductions in detoxication enzyme activities. The modest increases in susceptibilities, not exceeding ca. 4.5-fold, for C. rosaceana exposed to neem were achieved with subjects from a susceptible laboratory colony; a larger response could be expected for resistant field populations having elevated detoxication enzyme levels. Neem-based insecticides could be useful tools for the management of orchard pests that have developed resistance to synthetic neurotoxins. These findings also contribute to a better understanding of the numerous physiological changes that occur in larval Lepidoptera following exposure to neem extracts.     

Biology and control of the sciarid Lycoriella auripila Winn. (Diptera: Lycoriidae) in mushroom culture

The life-history of L. auripila, as a pest of mushrooms, is outlined and an explanation is given of the different types of damage caused. The most satisfactory control is based on the incorporation of insecticides in compost at spawning. Emulsions of chlorfenvinphos and diazinon at 50 ppm are lethal to young larvae for at least 7 weeks but granular formulations of the former must be applied at 100 ppm to achieve comparable results. Drenches of 0–01% malathion, applied at about 40 gal/1000 ft², are effective against larvae in the casing.     

Feed‐through insecticides for the control of the sand fly Phlebotomus argentipes

Three rodent feed‐through studies were conducted to evaluate the efficacy of insecticides to control Phlebotomus argentipes (Diptera: Psychodidae). The initial test evaluated diflubenzuron, eprinomectin, fipronil and ivermectin as feed‐through treatments in Rattus rattus (Rodentia: Muridae). In the preliminary trial, all four insecticides yielded 100% mortality of P. argentipes larvae within 20 days of exposure to treated rodent faeces. Based upon the initial results, fipronil was evaluated further as a feed‐through utilizing Bandicota bengalensis (Rodentia: Muridae). The B. bengalensis trial evaluated fipronil against both adult and larval sandflies at 250 p.p.m., 100 p.p.m. and 50 p.p.m. The results showed the fipronil treatment to have 100% efficacy against larvae up to 20 days post‐treatment and over 74% efficacy against adult sandflies presented with B. bengalensis faeces up to 10 days post‐treatment at all three dosage levels. The results of the three studies suggest that all four insecticides may be useful tools with which to control Leishmania vector populations.     

Suseptibility of the Predatory Mite,Neoseiulus californicus (Acari: Phytoseiidae), to Seven Insecticides

Susceptibility of the predatory mite, Neoseiulus californicus (McGregor) (Acari: Phytoseiidae), to the field rates of seven commercial insecticides was evaluated in laboratory bioassays. In direct treatments with lufenuron, novaluron, pyrifluquinazon, and sulfoxaflor, 82–88% of N. californicus adult females survived 168 h after exposure. Females exposed to these four insecticides produced 14–19% fewer eggs compared to females in the control group. Eclosion of eggs deposited by treated females was not affected. The percentage of eggs that hatched and larval survival following direct exposure to these four insecticides were not seriously reduced. A total of 72–82% of immature N. californicus survived on leaf discs with residues of lufenuron, novaluron, pyrifluquinazon, and sulfoxaflor matured to adults. Emamectin benzoate, lepimectin and spirotetramat were highly toxic to both N. californicus adult females and larvae. Based on these results, lufenuron, novaluron, pyrifluquinazon and sulfoxaflor are promising candidates for use in integrated pest management programs where N. californicus is a major natural enemy.     

Lethal effects of Gliocladium virens,Beauveria bassiana and Metarhizium anisopliae on the malaria vector Anopheles albimanus (Diptera: Culicidae)

Control of Anopheles albimanus, the main vector of malaria on the coast of the State of Chiapas, is based mainly on application of chemical insecticides, which has resulted in resistance to most registered insecticides. Strategies for biological control may provide sustainable alternatives. We report on the lethal effects of a native isolate of Gliocladium virens on An. albimanus larvae and adults, compared to that of strains of Beauveria bassiana and Metarhizium anisopliae. Conidial suspensions of G. virens, B. bassiana and M. anisopliae cultured on Sabouraud agar were tested in bioassays with An. albimanus larvae and adults. Mosquito larvae were more susceptible to all fungi, compared to adults. On early and late instar larvae, M. anisopliae showed the most pathogenic effect (LC₅₀ of 1.4×10⁵ conidia/mL in early instars; 1.1×10⁵ conidia/mL in late instars), followed by G. virens (LC₅₀ of 3.3×10⁵ conidia/mL in early instars and 3.5×10⁶ conidia/mL in late instars). Metarhizium anisopliae sensu lato and the native G. virens could be considered good choices for An. albimanus control in southern Mexico.     

Bacillus sphaericus Mtx1 and Mtx2 toxins co-expressed in Escherichia coli are synergistic against Aedes aegypti larvae

Mtx1 and Mtx2 are mosquitocidal toxins produced by some strains of Bacillus sphaericus during vegetative phase of growth. Mtx1 from B. sphaericus 2297 shows higher toxicity against Culex quinquefasciatus larvae than to Aedes aegypti larvae whereas Mtx2 from B. sphaericus 2297 shows lower toxicity against C. quinquefasciatus than to A. aegypti larvae. To test synergism of these toxins against A. aegypti larvae, mtx1 and mtx2 genes were cloned into a single plasmid and expressed in Escherichia coli. Cells producing both Mtx1 and Mtx2 toxins exhibited high synergistic activity against A. aegypti larvae approximately 10 times compared to cells expressing only a single toxin. Co-expression of both toxins offers an alternative to improve efficacy of recombinant bacterial insecticides. There is a high possibility to develop these toxins to be used as an environmentally friendly mosquito control agent.     

Non-target effects of neonicotinoid seed treatments; mortality of coccinellid larvae related to zoophytophagy

Third and fourth instar Harmonia axyridis Pallas larvae feed directly on corn seedlings; therefore they may be at risk if the seedlings are chemically treated. Neonicotinoids are broad-spectrum systemic insecticides that are frequently applied to corn seeds prior to planting to protect seedlings from early-season root and leaf-feeding. In this study, H. axyridis larvae were exposed for 360 min to corn seedlings that had been grown from seeds treated with either thiamethoxam or clothianidin. Neurotoxic symptoms (trembling, paralysis, and loss of coordination) were observed in 72% of the H. axyridis larvae; recovery following neurotoxic symptoms rarely occurred (7%). Starvation of larvae for 48 h prior to seedling exposure may increase tissue consumption; however, there was no difference in the number of starved or satiated larvae that displayed neurotoxic symptoms and died. Neurotoxic symptoms and larval mortality were significantly more likely to occur if larvae were exposed to seedlings grown from seeds treated with neonicotinoids compared to the control. In addition, clothianidin resulted in significantly greater larval mortality (80%) compared to thiamethoxam (53%). Symptoms and mortality were rarely observed if seedlings were not treated with a neonicotinoid seed treatment (3% and 3%, respectively). Coccinellids are abundant in agroecosystems in which neonicotinoid seed treatments are likely to be applied. Therefore, the use of neonicotinoids may have negative effects on these non-target species if early-season leaf-feeding occurs in the field.