西花蓟马抗噻虫胺种群对杀虫剂的交互抗性及机制

噻虫胺是具有内吸和触杀等多种作用方式的新烟碱类杀虫剂,常用于防治入侵害虫西花蓟马。为明确抗性风险,本文研究了西花蓟马抗噻虫胺种群对多种杀虫剂的交互抗性及其机制。经过45代筛选,西花蓟马对噻虫胺产生了高水平抗性(56.8倍)。生物测定结果表明: 西花蓟马高抗噻虫胺种群与噻虫嗪、吡虫啉、毒死蜱、三氟氯氰菊酯、甲维盐存在中等水平交互抗性(18.6>RR₅₀>11.3),对辛硫磷及灭多威具有低水平交互抗性,与溴虫腈和多杀菌素不存在交互抗性。胡椒基丁醚(PBO)与磷酸三苯酯(TPP)对杀灭西花蓟马抗噻虫胺种群(CL)、云南田间种群(YN)和敏感种群(S)均有显著增效作用。西花蓟马抗噻虫胺种群细胞色素P₄₅₀含量(3.6倍)、细胞色素b₅含量(2.9倍)及O-脱甲基酶活性(4.9倍)和羧酸酯酶活性(2.5倍)均显著高于敏感种群,表明多功能氧化酶及羧酸酯酶活性增强是西花蓟马对噻虫胺产生抗性的重要机制。     

西花蓟马抗辛硫磷种群的抗性机制及交互抗性

为明确西花蓟马对辛硫磷的抗性风险,研究了西花蓟马抗辛硫磷种群对其他杀虫剂的交互抗性及其对辛硫磷的抗性机制.交互抗性测定结果表明,西花蓟马抗辛硫磷种群对辛硫磷与毒死蜱、高效氯氟氰菊酯和灭多威存在中等水平的交互抗性,对溴虫腈、吡虫啉、甲维盐和多杀菌素存在低水平交互抗性,对啶虫脒和阿维菌素不存在交互抗性.酶抑制剂与辛硫磷的增效剂测定结果表明,胡椒基丁醚(PBO)、三丁基三硫磷酸酯(DEF)和磷酸三苯酯(TPP)对西花蓟马抗辛硫磷种群(XK)、田间种群(BJ)和敏感种群(S)均起到了显著的增效作用(P＜0.05),马来酸二乙酯对西花蓟马抗辛硫磷种群和敏感种群增效作用均不显著,但对田间种群增效作用显著(P＜0.05).生化测定发现:除田间种群西花蓟马乙酰胆碱酯酶活性提高不显著外,西花蓟马抗辛硫磷种群和田间种群的细胞色素P450含量(2.79和1.48倍)、细胞色素b5含量(2.88和1.88倍)及O-脱甲基酶活性(2.60和1.68倍)、羧酸酯酶活性(2.02和1.61倍)和乙酰胆碱酯酶活性(3.10倍)均显著高于敏感种群(P＜0.05);谷胱甘肽-S-转移酶酶活性也有一定程度提高(1.11和1.20倍),但不显著(P＞0.05).表明其体内解毒代谢酶和靶标酶活性提高是西花蓟马对辛硫磷产生抗性的重要原因.     

西花蓟马对吡虫啉的抗性机制及交互抗性研究

为了解西花蓟马Frankliniella occidentalis(Pergande)对吡虫啉的抗性风险,本文就吡虫啉的交互抗性和抗性机制(增效剂和酶活性)进行了研究。结果表明,经过35代筛选,西花蓟马对吡虫啉的抗性上升到21.26倍。西花蓟马对吡虫啉与阿维菌素和甲维盐存在中等水平交互抗性,与氯氟氰菊酯、灭多威和毒死蜱存在低水平交互抗性。增效剂试验表明,三丁基三硫磷酸酯(DEF)、磷酸三苯酯(TPP)和马来酸二乙酯(DEM)具有显著增效(SR50,DEF=6.38,SR50,TPP=5.52,SR50,DEM=1.60,P<0.05)。生化测定表明:抗吡虫啉品系西花蓟马的羧酸酯酶(5.06倍)和谷胱甘肽S-转移酶酶活性(1.63倍)均显著(P<0.05)高于敏感品系,表明羧酸酯酶和谷胱甘肽S-转移酶酶活性的提高是西花蓟马对吡虫啉产生抗药性的主要原因。     

北京和云南地区西花蓟马对多杀菌素类药剂产生抗药性

【目的】了解我国西花蓟马Frankliniella occidentalis(Pergande)对多杀菌素等药剂的抗药性现状,为西花蓟马的有效防治提供参考。【方法】2011—2015年,采用叶管药膜法对北京和云南地区西花蓟马进行了抗药性监测。【结果】与室内敏感品系相比,北京地区西花蓟马田间种群对多杀菌素的抗性倍数达到80~150倍,对乙基多杀菌素抗性倍数高达7 730倍,对乙基多杀菌素的LC50值5年间最高增加了258倍,对甲维盐和噻虫嗪具有中等水平抗性,对阿维菌素和虫螨腈处于敏感或低水平抗性;昆明地区西花蓟马对乙基多杀菌素的抗性水平达到305倍的极高抗水平,对其它药剂相对敏感或处于低水平抗性。【结论】北京和云南地区西花蓟马对多杀菌素类药剂已经产生高水平抗药性,应密切关注抗性发展动态,同时进一步研究其抗性机制。     

西花蓟马田间种群对常用杀虫剂的抗性现状及防治对策

【目的】西花蓟马Frankliniella occidentalis在中国是一种严重危害温室蔬菜的入侵害虫。本研究旨在了解该害虫在中国的抗药性现状,为防治该害虫提供理论支持。【方法】采用Munger cell法测定了北京,山东寿光和青岛以及云南晋宁和呈贡等5个地区西花蓟马田间种群对多杀菌素、毒死蜱、阿维菌素、甲维盐、氟氯氰菊酯、溴虫腈、灭多威、吡虫啉和啶虫脒9种杀虫剂的抗药性水平,同时利用这些田间种群测定了多功能氧化酶抑制剂胡椒基丁醚（PBO）、谷胱甘肽S 转移酶抑制剂顺丁烯二酸二乙酯（DEM）和羧酸酯酶抑制剂三丁基三硫磷酸酯（DEF）对多杀菌素、吡虫啉和甲维盐的增效作用。【结果】生物测定结果表明,北京、晋宁及呈贡种群分别对多杀菌素产生了34.45, 47.45和64.45倍的高水平抗性;晋宁种群对灭多威和甲维盐分别产生了16.58和11.03倍的中等水平抗性;呈贡种群对甲维盐、啶虫脒、吡虫啉、阿维菌素、溴虫腈分别产生了24.17, 21.69, 20.05, 16.45和10.31的中等水平抗性;青岛种群对啶虫脒和吡虫啉产生了17.70和12.49倍的中等水平抗性;寿光种群没有对任何杀虫剂产生高等或中等水平抗性。增效剂生物测定结果表明,对于吡虫啉和甲维盐,多功能氧化酶抑制剂PBO在所有田间种群上均有显著的增效作用。谷胱甘肽S-转移酶抑制剂DEM在呈贡、寿光和青岛种群中对吡虫啉存在显著增效作用;在北京、呈贡和寿光种群中,DEM对甲维盐存在显著增效作用。羧酸酯酶抑制剂DEF在呈贡、晋宁和青岛种群中对吡虫啉存在显著增效作用;在北京、呈贡和晋宁种群中,DEF对吡虫啉存在显著增效作用。但所有增效剂在各田间种群中对多杀菌素均无显著增效作用。【结论】结果提示：在使用多杀菌素防治西花蓟马时,应与其他杀虫剂轮换使用;此外,可通过添加酶抑制剂来增强甲维盐和吡虫啉对西花蓟马的防效。     

番茄斑萎病毒对多杀菌素抗性西花蓟马发育繁殖和药剂敏感性的影响

【目的】西花蓟马Frankliniella occidentalis (Pergande)是重要的入侵害虫,是番茄斑萎病毒（TSWV）最有效的传播媒介,TSWV对西花蓟马的生长发育有一定的影响。多杀菌素是防治西花蓟马最有效的药剂之一,但已有田间西花蓟马对多杀菌素产生抗药性的报道。TSWV对抗性西花蓟马是否也有影响及程度如何尚不清楚。本研究通过对此问题进行深入研究,以期为进一步了解TSWV对西花蓟马的影响提供依据。【方法】应用特定年龄-龄期及两性生命表的方法,研究用番茄斑萎病毒处理和未处理的多杀菌素抗性和敏感西花蓟马种群的生物学特性;用叶管药膜法测定不同处理种群对3种药剂（多杀菌素、甲氨基阿维菌素苯甲酸盐和虫螨腈）的敏感性变化。【结果】对于抗性品系,TSWV处理后西花蓟马的发育历期缩短,雌成虫寿命和产卵量略高,但与对照组差异不显著(P＞0.05),内禀增长率（r）和净生殖率（R₀）分别为0.0433 d^-1和2.210,显著高于对照组（分别为0.0356 d^-1和1.972）（P ≤ 0.001）。对于敏感品系,TSWV处理后西花蓟马的发育历期缩短,雌雄成虫寿命均显著延长（P ≤ 0.001）,产卵量也略有提高,R₀为4.125,显著高于对照组（3.979）（P ≤ 0.001）。TSWV处理后敏感和抗性西花蓟马对多杀菌素的敏感性没有发生明显变化,对甲氨基阿维菌素苯甲酸盐和虫螨腈的敏感性显著降低。【结论】番茄斑萎病毒对多杀菌素敏感和抗性西花蓟马均有直接有利影响,病毒处理的西花蓟马发育历期缩短,繁殖能力增强,成虫寿命延长,对药剂的敏感性降低。     

西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪的抗性风险和抗性稳定性

为明确使用新烟碱类杀虫剂烯啶虫胺、噻虫胺和噻虫嗪防治入侵害虫西花蓟马的抗性风险及抗性稳定性,本研究采用芸豆浸药法对西花蓟马敏感种群初羽化雌成虫进行连续筛选获得抗性种群,根据抗性现实遗传力计算公式分析西花蓟马对上述3种杀虫剂的抗性风险,预测其抗性发展速度,并测定抗性稳定性。结果表明: 经过30代抗性筛选,西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪均达到高水平抗性(44.7、45.5和32.7倍)。西花蓟马对噻虫胺、烯啶虫胺和噻虫嗪的抗性发展速度依次降低,抗性现实遗传力分别为0.1503、0.1336和0.1258。对抗性种群在无选择压力下继续饲养10代,西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪的抗性水平均出现一定程度的下降,但均未能恢复到敏感性水平。抗性选育后,西花蓟马若虫与成虫对杀虫剂的敏感性差异显著缩小,西花蓟马敏感种群及抗性种群若虫对上述3种杀虫剂的敏感性显著高于成虫。西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪均存在高抗风险,噻虫嗪的抗性上升速度较慢且抗性稳定性最低。因此,在西花蓟马若虫期使用噻虫嗪有利于西花蓟马防治。     

吡虫啉胁迫对西花蓟马生长发育及种群性比的影响

为明确吡虫啉持续大量使用对外来入侵害虫西花蓟马Frankliniella occidentalis的影响。本研究通过毒力测定得到西花蓟马室内种群LC_(30)及LC_(50)的两种吡虫啉浓度162 mg/L和171 mg/L,在室内人工气候箱环境下研究了西花蓟马各虫态的发育历期、成虫寿命及性比等相关种群参数。结果表明,西花蓟马未成熟虫期的发育历期在经过吡虫啉处理后相比对照显著缩短(P0.05),且雌性西花蓟马的未成熟虫期明显短于雄性西花蓟马,其中以171 mg/L吡虫啉处理后西花蓟马的雌性种群未成熟虫期发育历期缩短最为明显(P0.05);西花蓟马雄性种群的寿命略长于雌性种群,且171 mg/L吡虫啉处理后雌雄虫寿命与对照相比均显著缩短(P0.05);吡虫啉处理后西花蓟马的平均产卵期低于对照,但单雌产卵量、日均产卵量均显著高于对照,171 mg/L吡虫啉处理后的单雌产卵量和日均产卵量最高(P0.05)。吡虫啉胁迫后,西花蓟马的雌性种群在整个种群中所占比例增速迅速上升,经过125 d(大约5代)之后,雌性种群已经占绝对优势,几乎取代雄性种群,即种群中90%以上均为雌性西花蓟马。西花蓟马雌性种群对吡虫啉的胁迫适应性强于雄性种群,吡虫啉胁迫是造成西花蓟马雌性种群比例迅速升高的主要原因之一,在吡虫啉浓度LC_(30)到LC_(50)胁迫的范围内,随着吡虫啉胁迫浓度的增加,种群中雌性种群所占比例增大。     

六种常见杀虫剂对西花蓟马和花蓟马的毒力测定

为了明确常用杀虫剂对入侵害虫西花蓟马和本地近缘优势种花蓟马的毒力作用,在室内采用菜豆浸渍饲喂法,分别测定了6种常用杀虫剂对云南昆明地区两种蓟马2龄若虫、雌虫及雄虫的毒力。结果表明:6种杀虫剂对西花蓟马和花蓟马2龄若虫和成虫的LC_(50)以6%乙基多杀菌素的最低,对西花蓟马和花蓟马2龄若虫和成虫LC_(50)值分别为:0.611、0.333 mg/L和1.731、1.202 mg/L;LC_(50)值以10%吡丙醚最高,对西花蓟马和花蓟马2龄若虫和成虫LC_(50)值分别为1238.005、845.819 mg/L和9037.110、4766.376 mg/L。6种杀虫剂对西花蓟马和花蓟马各虫态的毒力作用依次为:乙基多杀菌素阿维菌素溴氰菊酯啶虫脒吡虫啉吡丙醚,且所有供试杀虫剂对西花蓟马LC_(50)均大于花蓟马,对雌虫LC_(50)大于雄虫。说明了蓟马雌虫对杀虫剂的敏感度低于雄虫,且生物源杀虫剂的室内杀虫效果强于化学杀虫剂,是西花蓟马防治中优先选择的杀虫剂。     

西花蓟马对多杀菌素的抗性生化机制研究

在室内持续用多杀菌素对西花蓟马Frankliniella occidentalis(Pergande)进行抗性汰选,获得抗性倍数达到11 999倍的极高抗品系。该抗性品系对乙基多杀菌素和噻虫嗪具有显著的交互抗性,抗性倍数分别为53 718和84倍,对阿维菌素和毒死蜱的敏感性也有显著下降,交互抗性倍数分别为3.33和2.28倍,对虫螨腈无明显交互抗性。生化测定结果表明,抗性品系的羧酸酯酶、谷胱甘肽S-转移酶和多功能氧化酶的活力与敏感品系均无显著差异,推测抗性可能与靶标位点的敏感性降低有关。     

Application technique and irrigation method affect imidacloprid control of silverleaf whiteflies (Homoptera: Aleyrodidae) on poinsettias.

Subirrigation systems are increasingly used to water and fertilize greenhouse crops. They also appear to be well suited for the application of systemic pesticides. We conducted two studies to look at interactive effects ofimidacloprid application technique and irrigation method on plant uptake of imidacloprid and whitefly control. Drench applications of imidacloprid resulted in much higher concentrations in the leaves than applications to the bottom of pots after 14 d. However, imidacloprid efficacy in subirrigated plants was better if the imidacloprid was applied to the bottom of the pot than when an equal amount was applied as a drench. In drip-irrigated plants, imidacloprid efficacy was greater after a drench than after an application to the bottom of the pots. A second study showed that drench applications to drip-irrigated plants result in high imidacloprid concentrations in the bottom of the canopy, whereas bottom applications to subirrigated plants result in a more even distribution of imidacloprid throughout the plant. Surprisingly, the high leaf imidacloprid concentrations in the bottom layer of drip-irrigated plants did not result in improved whitefly control. Imidacloprid efficacy was better in subirrigated, bottom-treated plants than in drip-irrigated, drenched plants. Overall, results from these studies indicate that imidacloprid is very effective when applied to the bottom of subirrigated pots.     

Resistance to neonicotinoid insecticides and expression changes of eighteen cytochrome P450 genes in field populations of Bemisia tabaci from Xinjiang,China

The occurrence of Bemisia tabaci poses an increasingly serious threat to cotton and vegetable crops in Xinjiang, China. Currently, neonicotinoid insecticides are commonly used to control the insect, to which resistance is inevitable due to intensive use. However, the resistance status and mechanism of B. tabaci to neonicotinoid insecticides in Xinjiang are poorly understood. Cytochrome P450 monooxygenases represent a key detoxification mechanism in the neonicotinoid resistance of B. tabaci. In this study, the resistance level to imidacloprid and thiamethoxam was investigated using the leaf dipping method in five field populations of B. tabaci from Turpan (TP, two sampling sites), Shache (SC), Hotan (HT) and Yining (YN) in northern and southern Xinjiang. The expression changes of eighteen cytochrome P450 genes from the select B. tabaci populations were determined by real‐time fluorescence quantitative PCR (qPCR). The bioassay revealed that the five populations tested had developed moderate to high levels of resistance to imidacloprid (12.26–46.07‐fold), while the populations remained sensitive to thiamethoxam except for HT, which had a low level of resistance. The qPCR results showed that the expression levels of five P450 genes, CYP4G68, CYP6CM1, CYP303A1‐like, CYP6DZ7 and CYP6DZ4, were significantly higher in some resistant field populations than in the susceptible strain. Resistance to imidacloprid in field populations of B. tabaci might be associated with the increased expression of these five cytochrome P450 genes. The results are useful for further understanding the mechanism of neonicotinoid resistance and will contribute to the management of insecticide‐resistant B. tabaci in Xinjiang.     

Inheritance and synergism of resistance to imidacloprid in the Colorado potato beetle (Coleoptera: Chrysomelidae)

Colorado potato beetle, Leptinotarsa decemlineata (Say), adults and larvae collected from Long Island, NY, were 100.8 and 13.2 times more resistant to imidacloprid, respectively, compared with a susceptible strain. This high level of resistance appeared in only the third field season of imidacloprid use. Analysis of probit lines from F1 reciprocal crosses indicated that resistance to imidacloprid in adults was inherited autosomaly as an incompletely recessive factor. The degree of dominance of the resistance was -0.23 and -0.10, respectively, 3 and 7 d after treatment (incompletely recessive). The chi2 analysis of response ratio statistics from F1 x susceptible back crosses compared with a monogenic model suggested that more than one locus is responsible for resistance to imidacloprid. Synergism studies with piperonyl butoxide suggested that mixed-function oxidase mediated detoxification is responsible for the resistance to imidacloprid in adults. Synergism studies with S,S,S-tributyl phosphorotrithioate (DEF) indicated that esterase mediated detoxification may be an additional resistance factor. Mixed-function oxidase mediated detoxification is probably also one of the mechanisms of resistance to imidacloprid in larvae. Because the synergists used did not completely eliminate resistance in the resistant strain, there may be additional mechanisms involved. Refugia and crop rotation decrease the frequency of homozygous resistant genotypes and may be effective resistance management strategies, because of the recessive nature of the resistance.     

Evaluation of synergistic interactions between the Colorado potato beetle (Coleoptera: Chrysomelidae) pathogen Beauveria bassiana and the insecticides, imidacloprid, and cyromazine

Laboratory studies investigated the interaction between the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin and sublethal doses of the insecticides imidacloprid and cyromazine when applied to larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say). When second instars were fed potato leaf discs treated with sublethal doses of imidacloprid and a range of doses of B. bassiana, a synergistic action was demonstrated. Similar results were observed when larvae were sprayed directly with B. bassiana conidia and immediately fed leaf discs treated with imidacloprid. No synergistic interaction was detected when larvae were fed leaf discs treated with sublethal doses ofimidacloprid 24 h after application of B. bassiana conidia to larvae. However, a synergistic interaction was detected when larvae were fed leaf discs treated with imidacloprid and sprayed with B. bassiana conidia 24 h later. Although sublethal doses of both imidacloprid and the triazine insect growth regulator (IGR) cyromazine prolonged the duration of the second instar, only imidacloprid interacted with B. bassiana to produce a synergistic response in larval mortality. In leaf consumption studies, the highest dose of B. bassiana tested promoted feeding in inoculated second instars. Feeding was inhibited when larvae were fed foliage treated with sublethal doses of imidacloprid and significantly reduced when fed foliage treated with a sublethal dose of cyromazine. Starvation of larvae for 24 h immediately after B. bassiana treatment produced a similar result to the combined treatment of B. bassiana and imidacloprid and increased the level of mycosis when compared with B. bassiana controls. Imidacloprid treatment affected neither the rate of germination of B. bassiana conidia on the insect cuticle nor the rate at which conidia were removed from the integument after application. The statistical analysis used to detect synergism and the possible role of starvation-induced stress factors underlying the observed synergistic interactions are discussed.     

Managing Brachycaudis persicae (Homoptera: Aphididae) during peach orchard establishment

We investigated whether preplant root dips or postplant sprays or drenches could be used to control the black peach aphid, Brachycaudus persicae (Passerini), on young peach trees. In greenhouse trials, B. persicae died after being placed on peach trees that had their roots dipped in imidacloprid before planting. All rates (0.8, 1.6, and 3.2 ml/liter of water) tested controlled this aphid. B. persicae died faster on trees dipped in imidacloprid (1.6 ml/liter of water) for 5 min before planting than on trees dipped for 2 s. Aphids eventually died on all treated trees. Approximately 50% of untreated trees died from B. persicae infestations in greenhouse studies. Results from afield experiment show that peach trees root-dipped in imidacloprid before planting or drenched with imidacloprid after planting eliminate B. persicae infestations and prevent root colonization for at least 1 yr.     

Resistance irrelevant CYP417A2v2 was found degrading insecticide in Laodelphax striatellus

Cytochrome P450 monooxygenases (CYP s) usually overexpressed in resistant strain were found involved in oxidative detoxification of insecticides. In this study, an investigation was conducted to confirm if resistance irrelevant CYP s which were not overexpressed in resistant strain before, were capable of degrading insecticides. Three resistance irrelevant CYP s viz. CYP 417A2v2, CYP 425A1v2, and CYP 4DJ 1 from CYP 4 family of Laodelphax striatellus were randomly selected for experiments. CYP 417A2v2 and CYP 425A1v2 were found expressed successfully in Sf9 cell line while CYP 4DJ 1 was not expressed successfully and out of two expressed CYP s, only CYP 417A2v2 showed its efficient catalytic activity. For catalytic activity, three traditional model probe substrates and five insecticides were assayed. For the probe substrates screened, p‐nitroanisole and ethoxycoumarin were preferentially metabolized by CYP 417A2v2 (specific activity 3.76 ± 1.22 and 1.63 ± 0.37 nmol min^?1 mg protein^?1, respectively) and they may be potential diagnostic probes for this enzyme. Among insecticides, only imidacloprid was efficiently degraded by CYP 417A2v2. Incubation of imidacloprid with CYP 417A2v2 of L. striatellus and subsequent HPLC , LC ‐MS , and MS /MS analysis revealed the formation of imidacloprid metabolites, that is, 4′ or 5′hydroxy‐imidacloprid by hydroxylation. This result implies the exemption of CYP s character that it is not always, all the CYP s degrading insecticides being selected and overexpressed in resistant strains and the degrading CYP s without mutations to upregulate could be candidates during insecticide resistance evolution. This characterization of individual insect CYP s in insecticide degradation can provide insight for better understand of insecticide resistance development.     

Baseline susceptibility to imidacloprid and cross resistance patterns in Colorado potato beetle (Coleoptera: Chrysomelidae) populations

During 1995-1998, we tested 134 geographically discrete populations of Colorado potato beetle, Leptinotarsa decemlineata (Say), from the United States, Canada, Germany, France, and Poland for susceptibility to imidacloprid. Neonates were assayed on potato-based agar diet incorporated with imidacloprid and exposed on filter paper to esfenvalerate, azinphosmethyl, and carbofuran to characterize cross-resistance. In all 4 yr, Long Island populations were the most tolerant to imidacloprid, with LC50s ranging up to 29 times higher than the most susceptible populations. Responses to imidacloprid did not change significantly on farms where populations were assayed over time, except for those from Long Island, which doubled in overall tolerance to imidacloprid since 1995. Much of this tolerance was already present before imidacloprid was used on Long Island. Correlative analysis of the populations tested over the 4 yr indicated positive cross-resistance patterns with esfenvalerate and azinphosmethyl. This response was probably caused by preexisting metabolic and excretion mechanisms selected by previous exposure. There was no significant pattern of cross-resistance with carbofuran or bensultap. Regression slopes were also significantly negatively correlated with LC50 values for imidacloprid, indicating higher heterogeneity, which could lead in further resistance development. We discuss the relative sensitivity of diet-incorporated assays with neonates compared with other bioassay studies. Based on a pooled group of susceptible populations tested in 1995, a baseline LC50 of 0.39 ppm and a discriminating concentration of 8 ppm were suggested to detect early stages of resistance in "suspect" populations. We also suggest application strategies for imidacloprid that reduce selection pressure.     

Different utilizable substrates have different effects on cometabolic fate of imidacloprid in Stenotrophomonas maltophilia

Imidacloprid, the largest selling insecticide in the world, is more stable in soil, and its environmental residue and effects are attracting people's close attention. One of imidacloprid metabolism pathways was degraded to CO₂ through olefin imidacloprid pathway. Here, we report that sucrose as a utilizable substrate enhanced the cometabolism of imidacloprid by Stenotrophomonas maltophilia CGMCC 1.1788 to produce 5-hydroxy imidacloprid, whereas when succinate was used as a utilizable substrate, 5-hydroxy imidacloprid from imidacloprid was transformed to olefin imidacloprid, and the latter was further degraded. The hydroxylation of imidacloprid required NAD(P)H, whereas the dehydration of 5-hydroxy imidacloprid to form olefin imidacloprid required succinate rather than NAD(P)H. NADPH greatly favored the hydroxylation of imidacloprid more than NADH, and NADPH inhibited the dehydration of 5-hydroxy imidacloprid to olefin imidacloprid, but NADH did not. Therefore, sucrose may be metabolized through hexose monophosphate pathway to produce mainly NADPH which participated in the hydroxylation of imidacloprid to 5-hydroxy imidacloprid and meanwhile inhibited the dehydration of 5-hydroxy imidacloprid to olefin imidacloprid, whereas succinate may be metabolized mainly through the tricarboxylic acid cycle to produce NADH which was involved in hydroxylation of imidacloprid to 5-hydroxy imidacloprid but did not inhibit the dehydration of 5-hydroxy imidacloprid to olefin imidacloprid. Our results have a significant meaning in further understanding the influence of different utilizable substrates on the cometabolic pathways and the fate of environmental imidacloprid.     

Identification of biochemical markers linked to neonicotinoid cross resistance in Bemisia tabaci (Hemiptera: Aleyrodidae)

The whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), is a serious pest in many cropping systems world-wide and occurs in different biotypes. The most widespread one is the B-type, whereas the Q-biotype is nowadays still mostly restricted to Southern Spain. Neonicotinoid cross-resistance is known at a high level in Q-types from Spain and individual samples collected in Italy and Germany. Now we detected for the first time high neonicotinoid cross-resistance in a B-type from Israel. Target site resistance to imidacloprid using [(3)H]imidacloprid in nicotinic acetylcholine receptor (nAChR) binding assays could not be detected in any of these highly resistant strains. The impact of metabolizing enzymes such as esterases, glutathione S-transferases, and cytochrome P450-dependent monooxygenases in neonicotinoid resistance was studied biochemically with artificial substrates. Monooxygenase activity was increased 2-3-fold in moderately resistant strains (RF approximately 30) and even 5-6-fold in highly resistant strains (RF approximately 1,000). Only monooxygenase activity correlated with imidacloprid, thiamethoxam and acetamiprid resistance and, therefore, monooxygenases seem to be the only enzyme system responsible for neonicotinoid resistance in B. tabaci Q- and B-types. The oxidative degradation of imidacloprid in resistant Q-type strains could be confirmed by metabolism studies of [(14)C]imidacloprid in vivo. Five-hydroxy-imidacloprid could be detected as the only main metabolite. The insecticidal activity and binding affinity to nAChR of this compound was 10 times lower than imidacloprid itself in B. tabaci.     

Imidacloprid-induced impairment of mushroom bodies and behavior of the native stingless bee Melipona quadrifasciata anthidioides

Declines in pollinator colonies represent a worldwide concern. The widespread use of agricultural pesticides is recognized as a potential cause of these declines. Previous studies have examined the effects of neonicotinoid insecticides such as imidacloprid on pollinator colonies, but these investigations have mainly focused on adult honey bees. Native stingless bees (Hymenoptera: Apidae: Meliponinae) are key pollinators in neotropical areas and are threatened with extinction due to deforestation and pesticide use. Few studies have directly investigated the effects of pesticides on these pollinators. Furthermore, the existing impact studies did not address the issue of larval ingestion of contaminated pollen and nectar, which could potentially have dire consequences for the colony. Here, we assessed the effects of imidacloprid ingestion by stingless bee larvae on their survival, development, neuromorphology and adult walking behavior. Increasing doses of imidacloprid were added to the diet provided to individual worker larvae of the stingless bee Melipona quadrifasciata anthidioides throughout their development. Survival rates above 50% were only observed at insecticide doses lower than 0.0056 μg active ingredient (a.i.)/bee. No sublethal effect on body mass or developmental time was observed in the surviving insects, but the pesticide treatment negatively affected the development of mushroom bodies in the brain and impaired the walking behavior of newly emerged adult workers. Therefore, stingless bee larvae are particularly susceptible to imidacloprid, as it caused both high mortality and sublethal effects that impaired brain development and compromised mobility at the young adult stage. These findings demonstrate the lethal effects of imidacloprid on native stingless bees and provide evidence of novel serious sublethal effects that may compromise colony survival. The ecological and economic importance of neotropical stingless bees as pollinators, their susceptibility to insecticides and the vulnerability of their larvae to insecticide exposure emphasize the importance of studying these species.