Effects of Sublethal Doses of Chlorfluazuron on Pupal-testis in the Common Cutworm, Spodoptera fitura （F,） （Lepidoptera： Noctuidae）

The effects of sublethal doses （LD10：1.00 ng larval-1; LD30： 3.75 ng larva-1） of chlorfluazuron on number and size of developmental spermatocytes in pupal-testis during spermatogenesis of the common cutworm, Spodoptera litura （F.） are described. Chlorfluazuron was applied topically to newly-ecdysed fifth-instar larvae of S. litura under laboratory conditions. During first day of pupation, spermatids were the highest （3,430 ± 7）, however, elongated spermatocytes with mature sperm were the lowest （292 ± 16） in number in pupal-testis compared with other developing spermatocytes. During last day of pupation, elongated spermatocytes with mature sperm were the highest （3,581 ± 3）, however, spermatogonia were the lowest （86± 4） in number in pupal-testis compared with other developing spermatocytes. The number of spermatogonia, primary and secondary spermatocytes, spermatids and elongated cysts with mature sperm were increased during pupal-developmental days in the controls. However, their numbers were significantly reduced in LD10- and even more significantly reduced in LD30-treated pupal-testis compared with controls. Moreover, eupyrene sperm bundles were not present in pupal-testis. During pupal-developmental days, spermatids were the smallest （4.1 ± 0.9 dm in μm） and secondary spermatocytes were the largest （31.0 ± 1.8 dm in μm） in size compared with other developing spermatocytes. The sizes of the same developing spermatocytes were significantly reduced in LD10-treated and more significantly reduced in LD30-treated pupal-testis of S. litura compared with controls.

Effects of Sublethal Doses of Chlorfluazuron on Embryogenesis in Spodoptera fitura

This paper describes effects of sublethal-doses of chlorfluazuron on morphology of embryogenesis when LD10 （1.00 ng/larva） or LD30 （3.75 ng/larva） was applied topically to newly-ecdysed fifth-instars of common cutworm, Spodoptera litura F.. Experiments were conducted on eggs of F1 generation laid by subsequent adults under laboratory conditions. Fecundity was significantly reduced by LD10 and more significantly reduced by LD30 compared with untreated females. Number of unfertilized-unhatched-eggs was not significantly affected by the treatments, Fertilized eggs revealed changes in external coloration of eggs. Embryonic development of S. litura was divided into eight embryonic stages. During stage 5 （12-16 h after oviposition）, in LD10-treated-egg, crescent-shaped-embryo was affectedly differentiated. During the same stage, it was seriously affected in LD30-treated-egg. During stage 7 （32-64 h after oviposition）, in LD10-treated-egg, organogenesis was established, but body structures were affectedly developed. In LD30-treated-egg, body structures were more affectedly developed during the same stage. It is concluded that embryonic developmental stages were affected by LD10 and more seriously affected by LD30. The effects of residualchlorfluazuron on embryogenesis are thought to be one of the factors responsible for the reduction in fertility and hatchability caused by sublethal doses of chlorfluazuron in S. litura.

Effects of Chlorfluazuron Sublethal Doses on Spermatogenesis of Adult Common Cutworm, Spodoptera fitura （F.） （Lepidoptera： Noctuidae）

Chlorfluazuron is an insect growth regulator for controlling the major insect pests in crops. This paper describes the effects of sublethal doses （LD10：1.00 ng/larva; LD3o： 3.75 ng/larva） of chlorfluazuron on spermatogenesis in the adults of the common cutworm, Spodoptera litura （F.）. Chlorfluazuron was applied topically to the newly-ecdysed fifth-instars ofS. litura under laboratory conditions. The secondary spermatocytes, spermatids, elongated spermatocytes with mature sperm and eupyrene sperm bundles were present in testis of newly emerged, 1 and 2 day-old adults, but gradually decreased in their numbers in the controls. However, the spermatogonia were not present in newly emerged, 1 and 2 day-old adults. Moreover, primary spermatocytes were present （174 ± 4.0） in newly emerged adults, but not found in 1 and 2 day-old adults. In the testes of newly emerged LD10-treated adults, the numbers of eupyrene and apyrene sperm bundles were significantly reduced compared with controls, and even significantly reduced in LD30-treated adults compared with LDlo, however, their ratios were not affected. Moreover, the pattern of spermatogenesis was the same in LD10- and LD30-treated male adults compared with controls. Furthermore, numbers of developing spermatocytes were significantly reduced in the LD10 compared with controls, and even significantly reduced in LD30-treated male adults compared with LD10. In newly emerged adults, the width and length of elongated spermatocytes with mature sperm were significantly reduced in LDlo- and more significantly reduced in LD30-treated compared with controls of S. litura. In vas deferens of untreated pre- and newly emerged adult male, both eupyrene and apyrene sperm bundles were found, however, in treated pre-adult male, only apyrene sperm bundles and in treated newly emerged adult male, fe, w eupyrene and numerous apyrene sperm bundles were found. It is concluded that spermatogenesis was significantly affected by chlorfluazuron.

Effects of Sublethal Doses of Chlorfluazuron on Ovarioles in the Common Cutworm, Spodoptera Litura （F.） （Lepidoptera： Noctuidae）

Chlorfluazuron is an insect growth regulator for controlling the major insect pests in crops. This paper describes the effects of sublethal doses （LD10：1.00 ng/larva; LD30：3.75 ng/larva） of chlorfluazuron on ovarioles during ovarian development and oogenesis of common cutworm, Spodoptera litura （F.）. It is a significant pest of cotton, tobacco and miscellaneous vegetables. The LDlo or LD3o was applied topically to the newly-ecdysed fifth-instars of S. litura under laboratory conditions. In controls, on 2nd day of adult emergence, the cell density in germarium was 1636 ± 9.17 nuclei/mm^2. It increased until 3rd day, i.e., 1829 ± 8.87 nuclei/mm^2 and decreased thereafter. In the LDloor LD-3o-treated insects, the patterns of the cell density was the same as in controls but values were significantly （P 〈 0.05） decreased in the LD10- and more in the LDa0-treated females compared with controls. In control, ovaries were normal type and had mature ova with an occasional one or two being absorbed. In LD10-treated females, ovaries had spaces in ovarioles and absorption of ova. In LD30-treated females, besides the spacing and absorption, sometimes only immature （germarium） ova were found.

两种昆虫生长调节剂类杀虫剂对甜菜夜蛾幼虫储存蛋白含量的影响

通过阴离子交换、凝胶过滤等方法对甜菜夜蛾5龄幼虫储存蛋白进行纯化,采用SDS-PAGE、ELISA等方法测定甜菜夜蛾储存蛋白动态变化,研究两种昆虫生长调节剂类杀虫剂甲氧虫酰肼和氟啶脲在亚致死剂量下对储存蛋白含量的影响。结果表明:甜菜夜蛾幼虫储存蛋白亚基分子量为85.1 ku。在5龄幼虫血淋巴中储存蛋白质量浓度随日龄逐渐增加,于5龄第3天达到最大值30.76 mg∕mL。低剂量的甲氧虫酰肼能够提高幼虫血淋巴中储存蛋白的浓度,处理48 h和72 h后,LC10处理组的储存蛋白质量浓度分别为19.27 mg∕mL和45.21 mg∕mL,显著高于对照组。高剂量的甲氧虫酰肼对甜菜夜蛾储存蛋白存在着明显的抑制作用,LC40处理组在72 h后,血淋巴中储存蛋白浓度仅为对照组的48.47%。氟啶脲LC10、LC40两个处理组的储存蛋白浓度在各个处理时间均低于对照组,特别是在处理48 h和72 h后,血淋巴中储存蛋白浓度显著降低,且氟啶脲浓度越高,影响越显著,LC40处理组在72 h后,血淋巴中储存蛋白浓度仅为对照组的26.77%。研究结果可为进一步揭示两种杀虫剂对甜菜夜蛾幼虫的毒力作用提供理论依据。

10%氟铃脲饵剂防治台湾乳白蚁的效果观察

目的 通过现场试验,明确10%氟铃脲饵剂对台湾乳白蚁的灭治效果。方法 在确定台湾乳白蚁巢群大体数量的基础上,给每巢台湾乳白蚁施以不同剂量的饵剂,然后观察巢群的存活情况。结果 10%氟铃脲饵剂对栖息在树木内的台湾乳白蚁巢群具有良好的灭杀效果,每巢施用饵剂100~200 g时,可在28~42 d(平均33.7 d)内将整巢台湾乳白蚁杀灭。结论 10%氟铃脲饵剂可在较短的时间里杀灭台湾乳白蚁巢群,建议对其开展进一步的研究,以促进它在白蚁防治中的应用。

梨中除虫脲、氟虫脲、氟啶脲的残留分析方法

建立了同时分析梨中除虫脲、氟虫脲、氟啶脲3种农药的残留分析方法。样品以丙酮提取,二氯甲烷萃取,中性氧化铝柱净化,高效液相色谱(HPLC)测定。3种农药的检测限在0.6607～0.8809ng之间,最低检出质量分数在0.008～0.012mg/kg之间,平均添加回收率在83.81%～97.52%之间。该方法的准确性、精确性以及灵敏度均达到农药残留分析的要求。

小菜蛾对定虫隆抗性种群的选育及交互抗性研究

用昆虫生长调节剂（ｉｎｓｅｃｔｇｒｏｗｔｈｒｅｇｕｌａｔｏｒ，简称ＩＧＲ）定虫隆（ｃｈｌｏｒｆｌｕａｚｕｒｏｎ）对源自深圳田间的小菜蛾（ＳＺＳ）Ｐｌｕｔｅｌａｘｙｌｏｓｔｅｌａ（Ｌ）在室内进行抗性种群选育，经过８代饲养和６次药剂汰选，获得抗性种群（ＣＨＲ），与相对敏感种群ＳＺＳ比较，抗性指数（ＲＩ）为２３７８倍。ＣＨＲ种群在去除选择压力条件下饲养５代，抗性逐渐下降。抗性汰选前后分别测定了１０种药剂的剂量死亡毒力回归线，发现ＣＨＲ抗性种群对三氟氯氰菊酯、氯氰菊酯、辛硫磷、喹硫磷、灭多威、磺胺脲类衍生物杀螨隆、微生物杀虫剂Ｂｔ和齐墩螨素无明显交互抗性，抗性倍数为０４～１７；对两种沙蚕毒素类杀虫剂杀螟丹和杀虫丹的敏感性却有所上升，有负交互抗性趋势。活体增效剂试验表明，增效醚（ＰＢＯ）和三苯基磷酸酯（ＴＰＰ）对定虫隆均有一定的增效活性，ＰＢＯ的增效比最高为２９倍，能够完全抑制对定虫隆的抗性，说明多功能氧化酶可能是主导抗性机制之一。

球孢白僵菌对斜纹夜蛾高毒菌株筛选与制剂的研发

斜纹夜蛾Spodoptera litura是一种危害极大的农业害虫,为了探索1条可持续、无污染的新型防治方法,我们通过标准喷塔法从12株球孢白僵菌Beauveria bassiana中筛选出1株对斜纹夜蛾2龄幼虫高毒力的菌株———球孢白僵菌Bb2860菌株。在喷雾浓度为（2592±275）孢子.mm-2的高浓度下,菌株Bb2860对斜纹夜蛾2龄幼虫在第7d的校正死亡率达82.43%±6.03%。不同制剂的室内生测试验表明,孢子水悬液和2种油剂（分别为不含稳定剂和含有稳定剂）在中浓度319~529孢子.mm-2的喷雾浓度下,7d后对斜纹夜蛾2龄幼虫的累计死亡率分别为54.8%7、0.0%和76.7%;菌药协同作用为86.8%。另外,于2009年9月在安徽马鞍山进行自然条件下对花椰菜叶上斜纹夜蛾幼虫的田间小区药效试验。处理9d后,纯菌剂的相对防效为77.64%;低剂量和常规用量5%氟啶脲（抑太保）的相对防效分别为49.02%和89.70%;菌药混用的相对防效为91.13%。本研究显示出利用球孢白僵菌制剂及其与低剂量抑太保混用防治斜纹夜蛾的良好应用前景。

小菜蛾对定虫隆的抗性生化机制初探

以定虫隆汰选的抗性指数为２３７８倍抗性种群（ＣＨＲ），并以上海田间种群（ＳＨＲ，对定虫隆的抗性指数为９１６倍）作为参比种群，研究小菜蛾Ｐｌｕｔｅｌａｘｙｌｏｓｔｅｌａ（Ｌ）对定虫隆的抗性机制，结果表明：ＣＨＲ种群多功能氧化酶Ｏ脱甲基活力比相对敏感种群（ＳＺＳ）提高１倍，而ＳＨＲ种群多功能氧化酶Ｏ脱甲基活力虽有所提高，但与ＳＺＳ种群相比差异并不显著；ＣＨＲ和ＳＨＲ种群羧酸酯酶比活力均明显高于ＳＺＳ种群，分别为ＳＺＳ种群的４６１和２１８倍，且ＣＨＲ种群的Ｋｍ仅为ＳＺＳ种群的１／２０；酸性磷酸酯酶和碱性磷酸酯酶比活力种群间无明显差异；ＣＨＲ种群的几丁质酶活力降低４８％，酚氧化酶活力降低６０％。说明小菜蛾对定虫隆的抗性机制具有多因子性，多功能氧化酶解毒代谢能力的提高可能是主导抗性机制之一，羧酸酯酶、几丁质酶和酚氧化酶也参与了小菜蛾对定虫隆的抗性。

10%氟啶脲水悬浮剂润湿分散剂的筛选

采用流点法对10%氟啶脲水悬浮剂的润湿分散剂进行了初步筛选,在流点法筛选结果的基础上利用激光粒度仪测定粒径的方法对分散剂进行进一步的筛选。结果表明,农乳602、分散剂NNO、550S均能加工成流动性好的氟啶脲水悬浮剂,采用优化组合法和粘度法最终确定了10%氟啶脲水悬浮剂的润湿分散剂:农乳602和550S的比例为4:1,用量为0.8%。

广西小菜蛾对定虫隆的抗药性监测

从1997年下半年起，广西开始使用定虫隆防治小菜蛾。不同年份田间抗药性监测结果表明：2000年以前，小菜蛾对定虫隆仍敏感；2001～2002年小菜蛾对定虫隆的抗性指数为8．76～9．75，属低水平抗性；2003～2004年小菜蛾对定虫隆的抗性指数为20．41～21．25，已达到中抗水平；2002～2003年广西田间不同小菜蛾种群对定虫隆的抗性指数在11．43～28．14之间，达到中等水平抗性。在条件基本相同的情况下，田间药效试验结果为：2000年以前，5％抑太保乳油1000～2000倍液，药后7d对小菜蛾的防效均在90％以上，2001年只有1000倍液的防治效果达到90％以上，2002～2003年防效下降很快，到2004年，柳州、南宁试验点1000倍液的防治效果均低于70％，显著低于前几年的防效。抗性监测与田间药效变化的结果基本一致。

高效液相色谱法测定萝卜中定虫隆残留量

建立了快速测定萝卜中定虫隆残留方法。样品经乙腈提取,用高效液相色谱仪进行测定。实验结果表明,方法的最低检出量为1×10-9g,最低检出质量分数为0.02mg/kg,在0.1、0.5、1.0mg/kg三种添加水平下,添加回收率范围为85.16%～95.29%,变异系数范围为3.00%～7.53%。该分析方法快速、准确、灵敏,重现性好,适用于萝卜中定虫隆残留量的检测。

氟啶脲对甜菜夜蛾生长繁殖的亚致死效应

以LC_(10)、LC_(20)、LC_(20)和LC_(40)系列亚致死浓度的氟啶脲处理甜菜夜蛾2龄幼虫,研究其对甜菜夜蛾生长繁殖的影响。结果表明:氟啶脲对幼虫化蛹有明显抑制作用,随着氟啶脲浓度升高化蛹率降低,LC_(40)处理后的化蛹率仅为对照组的37.44%;亚致死浓度的氟啶脲可导致畸蛹率和成虫畸形率增加;亚致死浓度的氟啶脲对幼虫历期、蛹重、蛹期和羽化率等指标的影响不显著,并且性别间无明显差异。在繁殖性能方面,产卵前期随氟啶脲亚致死浓度升高而延长,LC_(40)处理组产卵前期为2.75 d;总产卵量随氟啶脲亚致死浓度升高而减少,LC_(40)处理组的总产卵量仅为对照组的42.78%;卵的孵化率随氟啶脲亚致死浓度的升高而降低,但无显著差异;产卵天数也随着亚致死浓度升高而缩短,对照组和LC_(20)、LC_(30)、LC_(40)处理组之间存在显著差异。

小菜蛾对抑太保的敏感性及其酰胺酶的活力

不同虫龄小菜蛾对抑太保的敏感性差异较大 ,其中 4龄是 1龄的 1 1 .6倍 .不同温度下抑太保对 3龄幼虫的 LC50 随温度的升高而迅速下降 ,1 0℃时为 1 2 43 .94mg·L- 1 ,3 5℃时则下降到 5 .0 6mg·L- 1 .酰胺酶的米氏常数 (Km)为 3 .65× 1 0 - 5nmol,最大反应速度 (Vm ax)为 0 .1 465 nmol· mg- 1 ·min- 1 .4龄幼虫酰胺酶的比活力和总活力随温度的升高而增大 .4℃饲养 1 5 d、1 0℃饲养 1 0 d及 2 5℃饲养条件下 ,4龄幼虫酰胺酶的比活力差异不大 ;但 3 5℃饲养 2 d与前 3个饲养条件相比 ,4龄幼虫酰胺酶的比活力差异较大 .酰胺酶的比活力和总活力均随虫龄的增长而增大 ,4龄与 1 -3龄相比酰胺酶的比活力差异较大 。

4.65%高氯·氟啶脲乳油防治菜青虫田间药效试验

通过 4 .6 5 %高氯·氟啶脲乳油对菜青虫的田间药效试验 ,结果表明 :4 .6 5 %高氯·氟啶脲乳油在药后第 1、3和 7d防效逐渐提高 ,至药后第 7d防效最好 ,6 0ml/ 6 6 7m2 最高防效为 96 76 % ,30ml/ 6 6 7m2 的最高防效为 93.5 5 % ,它兼有了高效氯氰菊酯的速效和氟啶脲的持效期长的优点。

超高效液相色谱测定柑橘中的虱螨脲、灭幼脲和氟啶脲

建立了柑橘中虱螨脲、灭幼脲和氟啶脲同时测定的超高效液相色谱（UPLC）分析方法。试样采用Qu ECh ERS法前处理,UPLC分析。结果表明：在0.05~0.50 mg/kg添加水平时,3种农药的回收率在82.5%~100.0%之间,相对标准差为0.3%~4.3%。方法的线性范围为0.05~1.0 mg/L,3种农药的相关系数均大于0.99,最低检测浓度均为0.05 mg/kg,能满足柑橘中3种农药残留检测的要求。

超临界流体萃取法测定定虫隆残留

将超临界流体萃取 (SupercriticalFluidExtraction ,SFE)技术与高效液相色谱分析相结合 ,得到了特异性杀虫剂定虫隆的残留分析方法。SFE对定虫隆的萃取条件为 :压力 3 0 0 0PSI ,温度75℃ ,静态萃取时间 2 0min ,动态萃取CO2 流量 50ml,改性剂 0 .0 8ml/g(甲醇 ) ,收集溶剂甲醇 ,萃取样品可直接用于高效液谱分析 ,条件为检测器 :紫外二极管阵列检测器 (UV2 54nm) ,色谱柱C18键合柱 ,流动相甲醇 /水 (80∶2 0 ,V/V) ,流速 1ml/min ,柱温 2 5℃ ,保留时间 :18.2 85min。该定虫隆残留分析方法回收率为 10 0 .94%。

小菜蛾过氧化氢酶和过氧化物酶与耐药性的关系

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