Induction of heat shock protein genes by chlorfenapyr in cultured cells of the cabbage armyworm,Mamestra brassicae

Effects of structurally different insecticides, permethrin, chlorfluazuron, chlorfenapyr, prothiofos, methomyl, and thiocyclam on expression of hsp90, hsp70, hsp20.7, and hsp19.7 were examined using cultured cells of the cabbage armyworm, Mamestra brassicae. Significant induction of hsp90, hsp70, hsp20.7, and hsp19.7 expression was observed in response to chlorfenapyr. No induction was observed for the remaining five insecticides. The chlorfenapyr-induced expression was time-dependent and concentration-dependent. A significant reduction in expression levels of hsp70, hsp20.7, and hsp19.7 was observed in the time course when the cells exposed to chlorfenapyr were transferred to chlorfenapyr-free medium. These results suggest that hsp70, hsp20.7, and hsp19.7 might be useful to assess cellular distress or injury by chlorfenapyr.     

Stressor-specific induction of heat shock protein 70 in the freshwater prawn Macrobrachium malcolmsonii (H. Milne Edwards) exposed to the pesticides endosulfan and carbaryl

Agricultural run off that is contaminated with pesticides enters water bodies, thereby polluting the aquatic environment. The sensitivity of the freshwater prawn, Macrobrachium malcolmsonii, to such pesticides is well-documented. However, the stress response to sublethal concentrations of pesticides has scarcely been investigated. In the present study, the effect of two different sublethal concentrations (1/5th and 1/10th of LC₅₀ value) of endosulfan and carbaryl on the synthesis of hsp70 in various tissues of the prawn M. malcolmsonii was evaluated by Western blotting analysis. Among the tissues analyzed (gill, skeletal muscle, and hepatopancreas), induction of hsp70 synthesis was recorded only in the gill tissue of the prawns that had been exposed to either the high or low sublethal concentration of endosulfan. Induction of synthesis of hsp70 was not observed in any of the tissues sampled from the prawns that had been exposed to either the low or the high sublethal concentration of carbaryl. Pesticides interfere with the hydrophobic core of protein, thus leading to denaturation of the protein by exposing the lipophilic moieties to the aqueous environment. Specific induction of hsp70 in the gill possibly occurred since the gill is the route of exposure for pesticides in the prawns. The absence of induction of hsp70 synthesis in prawns that had been exposed to carbaryl suggests that induction of hsp70 synthesis is stressor-specific.     

甜菜夜蛾敏感品系的获得及其对12种杀虫剂的敏感基线

以高效氯氰菊酯为筛选药剂,通过室内单对汰选获得了甜菜夜蛾敏感品系,并采用浸叶法和点滴法测定该敏感品系对甲氨基阿维菌素苯甲酸盐、氯虫苯甲酰胺等12种常用药剂的敏感基线。浸叶法测得该品系对供试药剂的敏感性由高到低依次为:甲氨基阿维菌素苯甲酸盐(LC50为0.0340 mg/L)、茚虫威、氟啶脲、甲氧虫酰肼、氯虫苯甲酰胺、氟虫双酰胺、高效氯氰菊酯、虫酰肼、多杀菌素、毒死蜱、虫螨腈、灭多威(124.0482mg/L)。点滴法测得12种杀虫剂LD50由小到大依次为:高效氯氰菊酯、茚虫威、甲氧虫酰肼、甲氨基阿维菌素苯甲酸盐、虫酰肼、毒死蜱、多杀菌素、氯虫苯甲酰胺、虫螨腈、氟虫双酰胺、灭多威、氟啶脲。结果表明,该敏感基线可用于甜菜夜蛾的抗药性监测。     

Deletion of Cyp6d4 does not alter toxicity of insecticides to Drosophila melanogaster

Cytochrome P450-dependent monooxygenases are important in the activation and detoxification of numerous insecticides. In this study, a Drosophila melanogaster Cyp6d4 null mutant was used to determine the role of this P450 in insecticide metabolism. This null mutant was generated by imprecise excision of a mobile P element located upstream to the P450 gene Cyp6d4. Comparative analysis between the non-functional mutant and relevant control strains shows that Cyp6d4 does not appear to be involved in the metabolism of chlorfenapyr, cypermethrin, diazinon, imidacloprid, malathion, oxamyl, parathion, or pyrethrum extract, even though these insecticides are known to be activated or detoxified by P450-monooxygenases. No obvious abnormalities in development were seen in the Cyp6d4 null mutant, indicating that Cyp6d4 is not critical for the metabolism of vital endogenous substrates.     

六种杀虫剂对小地老虎的毒力及对土壤生物安全性评价

为筛选出高效安全的土壤处理杀虫剂,室内采用浸叶法比较了3种新杀虫剂与3种已登记的杀虫剂对小地老虎的毒力,同时用人工土壤法和密闭法分别测定了6种药剂对蚯蚓的急性毒性和对土壤呼吸强度的影响。结果显示,溴虫腈、茚虫威和甲维盐对小地老虎3龄幼虫的毒力明显高于3种已登记药剂毒死蜱、辛硫磷和高效氯氟氰菊酯;溴虫腈、茚虫威和甲维盐对蚯蚓的LC50分别为1320．80、1153．42和167．91mg／kg,均低于其它3种药剂,属于低毒级;供试杀虫剂中以溴虫腈对小地老虎和蚯蚓的毒力选择性最高,其毒力选择性比值（蚯蚓LC50／小地老虎LC50）为7115．23;6种药剂对土壤微生物均无危害性。     

Being exposed to low concentrations of pirimiphos-methyl and chlorfenapyr has detrimental effects on the mobility of Trogoderma granarium

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.     

虫螨腈对斜纹夜蛾细胞的毒力及对线粒体膜电位的影响

以MTT法测定了虫螨腈等杀虫剂对斜纹夜蛾(Spodoptera litura,SL)细胞的毒力,并用流式细胞仪测定了虫螨腈对SL细胞线粒体膜电位的影响。结果表明: 100 μg/mL的氟虫腈、丁醚脲、虫螨腈处理SL细胞24 h后,对细胞增殖的抑制率依次为14.57%、22.91%、 52.53%;虫螨腈对SL细胞24和48 h的IC50值分别为57.56和2.60 μg/mL;50、100、200 μg/mL虫螨腈处理细胞24 h后,与对照相比,SL细胞线粒体膜电位分别降低46.87%、57.17%、73.31%,其下降值与药剂浓度呈正相关。     

四种低毒杀虫剂对小菜蛾生长发育及繁殖的亚致死效应

为系统评价4种低毒杀虫剂对小菜蛾Plutella xylostella（L.）的综合控制作用,采用叶片浸渍法处理小菜蛾3龄幼虫,测定了虫酰肼、氟啶脲、茚虫威和虫螨腈的亚致死浓度（LC₂₅）对小菜蛾生长发育和繁殖力的影响。结果表明,4 种药剂亚致死浓度处理使小菜蛾3~4龄幼虫发育历期延长0.43~1.93 d,平均单头雌、雄蛹重分别减轻0.83~1.31 mg和0.19~1.09 mg ,幼虫的化蛹率及羽化率则分别降低11.2%~34.3%和21.7%~33.6%,以氟啶脲和茚虫威处理的影响较为显著。药剂处理后成虫平均产卵量减少30.15~60.21粒/雌,卵孵化率降低8.85%~19.65%,以虫酰肼效果较为明显;虫酰肼、氟啶脲、茚虫威均显著降低小菜蛾的交配成功率。     

Molecular cloning and mRNA expression of a ryanodine receptor gene in the cotton bollworm, Helicoverpa armigera

Ryanodine receptors (RyRs) are the targets of novel diamide insecticides. The cotton bollworm, Helicoverpa armigera, is one of the most important cotton pests in the world. In this study, we report the full-length RyR cDNA sequence (named as HaRyR) of H. armigera. The 16,083-bp contiguous sequence encoded 5, 142 amino acid residues, which shares 80% and 78% overall identities with its homologues in Nilaparvata lugens (NlRyR) and Drosophila melanogaster (DmRyR), respectively. All hallmarks of RyR proteins are conserved in the HaRyR, including the GXRXGGGXGD motif conserved in the Ca²⁺ release channels and four copies of RyR domain unique to RyR channels. The previously identified seven lepidopteran-specific RyR residues were also found in HaRyR (N⁴⁹⁷⁷, N⁴⁹⁷⁹, N⁴⁹⁹⁰, L⁵⁰⁰⁵, L⁵⁰³⁶, N⁵⁰⁶⁸ and T⁵¹¹⁹). An amino acid sequence alignment showed that the N-terminal region of HaRyR (residues 188–295) shared high sequence identity with NlRyR (94%) and DmRyR (92%), and moderate sequence identity (47–50%) with three rabbit RyR isoforms, while the short segment of the C-terminal transmembrane region of HaRyR (residues 4632–4676) exhibited moderate sequence identity with NlRyR (69%) and DmRyR (67%), and low sequence identity (19–28%) with three rabbit RyR isoforms. In addition, expression analysis of HaRyR revealed that the mRNA expression level in eggs was significantly lower than in third instar larvae, pupae and adults, and anatomical regulation of HaRyR expression was also observed with the highest expression level in head compared with thorax and abdomen. Our results lay a foundation for comprehensive structural and functional characterization of HaRyR and for understanding of the molecular mechanisms of toxicity selectivity of diamide insecticides among different species.     

北京地区粘虫对5种杀虫剂的抗药性

采用浸叶法测定了北京地区6个粘虫Mythimna separata(Walker)田间种群对5种不同类型杀虫剂的抗药性。结果表明:与相对敏感品系相比,6个田间种群对5种杀虫剂均表现出不同程度的抗性水平。其中,对氯虫苯甲酰胺(抗性倍数为1.314~4.213)、甲氨基阿维菌素苯甲酸盐(抗性倍数为1.000~4.385)和毒死蜱(抗性倍数为1.083~5.936)表现为敏感至低水平抗性;对虫螨腈(抗性倍数为1.355~20.80)和氯氟氰菊酯(抗性倍数为1.748~13.98)表现为敏感至中等水平抗性。因此,北京地区的粘虫防治应注重将氯虫苯甲酰胺、甲氨基阿维菌素苯甲酸盐和毒死蜱与虫螨腈或氯氟氰菊酯交替或轮换使用,以延缓抗药性的产生与发展。     

Methylparathion- and carbofuran-induced mitochondrial dysfunction and oxidative stress in Helicoverpa armigera (Noctuidae: Lepidoptera)

The cotton bollworm, Helicoverpa armigera is a polyphagous pest of several crops in Asia, Africa, and the Mediterranean Europe. Organophosphate and carbamate insecticides are used on a large-scale to control Helicoverpa. Therefore, we studied the effect of methylparathion and carbofuran, an organophosphate and carbamate insecticide, respectively, on oxidative phosphorylation and oxidative stress in H. armigera larvae to gain an understanding of the different target sites of these insecticides. It was observed that state III and state IV respiration, respiratory control index (RCI), and P/O ratios were inhibited in a dose-dependent manner by methylparathion and carbofuran under in vitro and in vivo conditions. Methylparathion and carbofuran inhibited complex II by ∼45% and 30%, respectively. Lipid peroxidation, H₂O₂ content, and lactate dehydrogenase (LDH) activity increased and glutathione reductase (GR) activity decreased in a time- and dose-dependent manner in insecticide-fed larvae. However, catalase activity was not affected in insecticide-fed larvae. Larval growth decreased by ∼64% and 67% in larvae fed on diets with 100 μM of methylparathion and carbofuran. The results suggested that both the insecticides impede the mitochondrial respiratory functions and induced lipid peroxidation, H₂O₂, and LDH leak, leading to oxidative stress in cells, which contribute to deleterious effects of these insecticides on the growth of H. armigera larvae, along with their neurotoxic effects.     

Sensitivity of the mitotic cells of barley (Hordeum vulgare L.) to insecticides on various stages of cell cycle

In this study, the genotoxic effects of insecticides in different phases of cell cycle were investigated in the mitotic cells of barley (Hordeum vulgare L.). The seeds of H. vulgare L. Var. Karan 4 were treated with different concentrations (0.05%, 0.1% and 0.5%) of insecticides alphamethrin (AM) and monocrotophos (MP) for 6 h after synchronization of cells in G₁, S and G₂ phase of cell cycle with the help of various presoaking durations (7 h, 17 h and 27 h.). Positive and negative control was run parallel in the form of ethyl methane sulphonate (EMS) and distilled water, respectively. The data indicate that higher dose of alphamethrin and monocrotophos produce toxicity, chromosomal aberrations and mitotic aberrations in Hordeum vulgare L. The present study indicates the genotoxic potential of the insecticides AM and MP and it also showed that S-phase of cell cycle was more sensitive compared to the G₁ and G₂ phases.     

Acute lethal and behavioral sublethal responses of two stored-product psocids to surface insecticides

BACKGROUND: The psocids Liposcelis bostrychophila Badonnel and L. entomophila (Enderlein) (Liposcelididae) are emerging pests of stored products. Although their behavior, particularly their high mobility, may contribute to the reported relatively low efficacy of insecticides against them, studies to investigate this have not been conducted. The present study aimed to assess the label rate efficacy of three commercial insecticides (β‐cyfluthrin, chlorfenapyr and pyrethrins) applied on concrete surfaces against L. bostrychophila and L. entomophila, and also their sublethal effect on the mobility of these species. RESULTS: The synthetic insecticides β‐cyfluthrin and chlorfenapyr showed high short‐term efficacy (LT₉₅≤15 h) against both psocid species, unlike the natural pyrethrins (LT₉₅ ≥ 4 days). Liposcelis bostrychophila was slightly more tolerant (≥1.2×) than L. entomophila to all three insecticides. Behavioral assays with fully sprayed and half‐sprayed concrete arenas indicated that the insecticides reduced the mobility of both species. Pyrethrins seem to elicit weak repellence in L. bostrychophila. CONCLUSION: β‐Cyfluthrin and chlorfenapyr were effective against both psocid species, but not pyrethrins. The mobility of both species does not seem to play a major role in the differential selectivity observed, but the lower mobility of L. bostrychophila may be a contributing factor to its higher insecticide tolerance. Published in 2008 by John Wiley & Sons, Ltd.     

氯虫苯甲酰胺和氟虫双酰胺对不同虫态甜菜夜蛾的毒力作用

采用浸叶法、点滴法、夹毒叶片法、成虫药膜法、饲喂毒营养液法测定了氯虫苯甲酰胺、氟虫双酰胺对甜菜夜蛾卵、2~4龄幼虫、成虫的活性和对4龄幼虫的拒食活性,以明确氯虫苯甲酰胺和氟虫双酰胺对不同虫态甜菜夜蛾的毒力作用。结果表明,两种杀虫剂均不具有杀卵活性,但可以明显降低初孵幼虫存活率;氯虫苯甲酰胺、氟虫双酰胺均对2龄幼虫活性最高,分别为3龄幼虫的1.50、6.17倍和4龄幼虫的22.25、44.95倍,对4龄幼虫胃毒活性分别为触杀活性的14.10、8.44倍;氯虫苯甲酰胺对甜菜夜蛾成虫毒力为氟虫双酰胺的25.53~34.19倍。氯虫苯甲酰胺、氟虫双酰胺对甜菜夜蛾幼虫表现出较强的拒食作用, 至处理后48 h拒食率达到最高, 分别为 82.73%、84.15%,显著高于对照药剂甲维盐各浓度处理,处理试虫虫体皱缩、体节缩短,体重增加率明显低于对照和甲维盐各处理。     

Resistance in field populations of <Emphasis Type="Italic">Amrasca devastans</Emphasis> (Hemiptera: Cicadellidae) to new insecticides in Southern Punjab,Pakistan

Cotton jassid, Amrasca devastans (Distant), is an economically important pest of various crops (i.e. cotton, okra, brinjal) and has the potential to become resistant against insecticides due to the intense use of insecticides in the region which entails high selection pressure. To monitor the resistance levels against recently used new chemistry insecticides (nitenpyram, spirotetramat, chlorfenapyr, fipronil, emamectin benzoate, sulfoxaflor, and flonicamid), four field populations of A. devastans collected from Khanewal, Multan, Muzaffargarh and Lodhran districts were examined by leaf dip bioassays. The results revealed the 32.95–136.47 fold resistance to nitenpyram, 23.03–56.74 fold to spirotetramat, 10.84–31.33 fold to chlorfenapyr, 1.20–9.43 fold to fipronil, 3.27–43.77 fold to emamectin benzoate, 0.66–2.81 fold to sulfoxaflor and 0.59–1.25 fold to flonicamid in different field populations, compared to the susceptible strain. Based on present study findings, it may be suggested to discontinue the use of nitenpyram, spirotetramat, chlorfenapyr and emamectin benzoate for which the populations were resistant and the rotational use of fipronil, sulfoxaflor and flonicamid for which the populations were susceptible. Proactive resistance management approaches such as judicious use of insecticides with proper dose selection, time of spray on early stages and correct method of application are necessary to prevent the development of resistance.     

Insecticide resistance in house flies from the United States: Resistance levels and frequency of pyrethroid resistance alleles

Although insecticide resistance is a widespread problem for most insect pests, frequently the assessment of resistance occurs over a limited geographic range. Herein, we report the first widespread survey of insecticide resistance in the USA ever undertaken for the house fly, Musca domestica, a major pest in animal production facilities. The levels of resistance to six different insecticides were determined (using discriminating concentration bioassays) in 10 collections of house flies from dairies in nine different states. In addition, the frequencies of Vssc and CYP6D1 alleles that confer resistance to pyrethroid insecticides were determined for each fly population. Levels of resistance to the six insecticides varied among states and insecticides. Resistance to permethrin was highest overall and most consistent across the states. Resistance to methomyl was relatively consistent, with 65–91% survival in nine of the ten collections. In contrast, resistance to cyfluthrin and pyrethrins + piperonyl butoxide varied considerably (2.9–76% survival). Resistance to imidacloprid was overall modest and showed no signs of increasing relative to collections made in 2004, despite increasing use of this insecticide. The frequency of Vssc alleles that confer pyrethroid resistance was variable between locations. The highest frequencies of kdr, kdr-his and super-kdr were found in Minnesota, North Carolina and Kansas, respectively. In contrast, the New Mexico population had the highest frequency (0.67) of the susceptible allele. The implications of these results to resistance management and to the understanding of the evolution of insecticide resistance are discussed.     

Life-stage-dependent side effects of selected insecticides on <Emphasis Type="Italic">Trichogramma cacoeciae</Emphasis> (Marchal) (hymenoptera: Trichogrammatidae) under laboratory conditions

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is classified as one of the most harmful pest of tomato crops. Many species of predators and parasitoids including Trichogramma cacoeciae (Marchal) (Hymenoptera: Trichogrammatidae) are noted as potential candidates used for biological control of this pest. Therefore, the use of selective insecticides is critical to conserve and protect natural enemies in the field. This study assessed the side effects of insecticides on different development stages of T. cacoeciae under laboratory conditions. For this, eleven pesticides such as: Indoxacarb, spiromesifen, cyromazin, chlorfenapyr, cypermethrin, diafenthiuron, chlorantraniliprole, spinosad, azadirachtin, Bacillus thuringiensis (Bt) and virus HaNPV were tested. This study shows that indoxacarb, spiromesifen, chlorfenapyr, cypermethrin, diafenthiuron and spinosad had a negative effect on immature stages of Trichogramma. All insecticides residues on tomato leaves were found to be toxic to Trichogramma adults except azadirachtin, Bt and virus HaNPV. Therefore, the use of the tested natural products (azadirachtin, Bt and HaNPV) at the recommended doses is viable, having no negative impact on T. cacoeciae in tomato crops.     

Chlorfenapyr resistance in the spider mite Tetranychus urticae: stability, cross-resistance and monitoring of resistance

The two-spotted spider mite, Tetranychus urticae Koch, is a key pest of many agricultural crops. Studies of stability of resistance, cross-resistance relationships and monitoring of chlorfenapyr resistance were carried out with T. urticae to provide basic information necessary to define effective acaricide resistance management strategies for this pest. Chlorfenapyr resistance was shown to be stable in the absence of selection pressure under laboratory conditions. The activities of seven different acaricides against chlorfenapyr-resistant and -susceptible strains of T. urticae were evaluated. The results indicated possible positive cross-resistance between chlorfenapyr and the acaricides abamectin, propargite and etoxazole. No cross-resistance was detected for the acaricides milbemectin, fenpyroximate and diafenthiuron. A possible negatively correlated cross-resistance was observed between chlorfenapyr and spiromesifen. The evaluation of 21 T. urticae populations from several crops in the States of São Paulo, Mato Grosso, Goiás, and Bahia, in Brazil, indicated that the susceptibility of mites to chlorfenapyr was variable, with percentages of resistant mites ranging from 0.0 to 86.5%. The highest resistance frequencies were observed in ornamental plants in the State of São Paulo. Some populations from cotton and papaya also presented high frequencies of chlorfenapyr resistance. This is the first report on chlorfenapyr resistance in T. urticae on cotton and papaya in Brazil. Strategies for the management of acaricide resistance are discussed.