螺虫乙酯对斑马鱼的急性毒性及细胞凋亡的影响

螺虫乙酯作为一种结构新颖、作用机制独特和广谱高效的季酮酸类衍生物杀虫杀螨剂,在农业生产中普遍应用,随着用量的增加,对水生生物会造成一定的影响。采用经济合作与发展组织(OECD)方法在预试验基础上设螺虫乙酯2.5、2.97、3.54、4.20、5.00 mg/L 5个浓度梯度,研究其对斑马鱼的急性毒性,采用酚-氯仿萃取的方法提取DNA,并进行了优化,用DNA Ladder方法观察其细胞凋亡现象,用Giemsa染色观察其外周血细胞微核率,初步探索其致毒机制。结果表明,螺虫乙酯对斑马鱼72 h的LC50值为5.898 mg/L、96 h的LC50值为3.642 mg/L,属中等毒性;各浓度梯度螺虫乙酯药剂处理后的斑马鱼DNA出现梯状条带,螺虫乙酯染毒后对斑马鱼造成一定程度上的细胞凋亡现象;处理组微核率为(7.250±1.893)～(5.500±0.577),与对照组和溶剂组差异均达显著水平。     

螺虫乙酯对雌性斑马鱼的毒性及氧化应激效应

为评价新型季酮酸衍生物类杀虫剂螺虫乙酯(spirotetramat)对水生生物的毒性,以雌性斑马鱼为受试对象,研究了螺虫乙酯对其的急性毒性及氧化应激效应。急性毒性试验测得螺虫乙酯对雌性斑马鱼的96 h-LC50值为7.21 mg/L,属中等毒性。经不同浓度螺虫乙酯(0、36、360和1 800μg/L)处理28 d后,通过检测铜/锌超氧化物歧化酶(Cu/Zn-SOD)、锰超氧化物歧化酶(Mn-SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等抗氧化酶的活性,Cat、Gpx、Mn-Sod、Cu/Zn-Sod等基因的相对表达量,以及丙二醛(MDA)含量等指标的变化水平,研究了螺虫乙酯对斑马鱼肌肉组织的氧化应激效应。结果表明,螺虫乙酯可引起斑马鱼肌肉组织中CAT、GSH-Px活性和MDA含量,以及Cat、Mn-Sod、Gpx的相对表达量显著升高,但未发现螺虫乙酯对Cu/Zn-SOD活性及Cu/Zn-Sod的相对表达量有明显影响。研究表明,螺虫乙酯能够诱导雌性斑马鱼肌肉组织的氧化应激效应,并可能引起机体的氧化损伤。     

螺虫乙酯在黄瓜及其土壤中残留量检测方法及残留规律研究

[目的]本文旨在研究黄瓜和土壤中螺虫乙酯残留量的高效液相色谱测定方法,并于2014年用该方法测定湖南、浙江和广东三地螺虫乙酯在黄瓜及其土壤上的消解动态和残留规律试验研究的样品。[方法]样品用乙腈提取,高效液相色谱紫外检测器测定。[结果]结果表明：当添加螺虫乙酯在黄瓜及其土壤中的浓度为0.05mg/kg～1.0mg/kg时,平均添加回收率分别为95.75%～98.91%、89.37%～99.78%;变异系数分别为1.26%～2.89%、1.62%～1.99%,该方法对黄瓜和土壤中螺虫乙酯的最低检出浓度为 0.025mg/kg;在自然环境条件下,螺虫乙酯在湖南、浙江和广州三地的黄瓜和土壤中的半衰期分别为2.67～8.82d,1.95～6.19d。[结论]按推荐使用剂量对黄瓜施药,末次施药后第5天螺虫乙酯在三地的黄瓜和土壤中的残留量均未检出,黄瓜中螺虫乙酯残留量均低于瓜果类的MRL值(0.2mg/kg)。     

毒死蜱、吡虫啉、螺虫乙酯及其代谢物和苯醚甲环唑在梨中的残留消解动态

利用仪器分析方法和田间试验法，研究了毒死蜱、吡虫啉、螺虫乙酯及其代谢物和苯醚甲环唑在梨中的残留消解动态。样品经乙腈提取，氯化钠盐析净化，毒死蜱采用气相色谱-质谱联用仪检测，其他3种农药采用液相色谱-质谱联用仪检测，外标法定量。结果表明:毒死蜱、吡虫啉、螺虫乙酯及其代谢物和苯醚甲环唑4种农药在梨中的消解动态均满足一级反应动力学方程，半衰期分别为4.4、12.2、13.1和10.3 d。施药后7 d至收获期，4种农药在梨中的最终残留量均未超出中国国家标准中规定的最大残留限量值，按照推荐剂量及其操作规范在梨上施用是安全的。     

新型双向传导杀虫剂—螺虫乙酯对Q型烟粉虱的防治效果

为了筛选出防治Q型烟粉虱的有效药剂,研究了22.4%螺虫乙酯悬浮剂不同浓度、不同施药时间对烟粉虱卵、若虫的防治效果。螺虫乙酯和对照药阿维菌素均对Q型烟粉虱卵无效,螺虫乙酯对若虫具有非常高的杀虫活性,无论在卵期施药、若虫低龄期施药还是高龄期施药,防效均达95%以上,和对照药阿维菌素的防效无显著性差异。施药后20d接虫,防效大幅降低到21.6%以下,施药30d后接虫对若虫无效。根据烟粉虱的发育历期推算,表明螺虫乙酯持效期可达30d。     

Comparative analyses of the toxic effects of imidacloprid and spirotetramat on Frankliniella occidentalis (Pergande), the vector of tomato zonate spot virus: a laboratory assay

The tomato zonate spot virus (TZSV) is responsible for substantial yield losses in vegetables, tobacco and other cash crops in China in recent years. The western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera, Thripidae), is the main vector of TZSV in Yunnan Province. Because controlling the population of insect vectors is the most efficient way to prevent epidemics of viral diseases, we conducted laboratory assays to assess the potential of using spirotetramat as an alternative to imidacloprid in controlling F. occidentalis. Our results demonstrated that spirotetramat was significantly more lethal than imidacloprid to F. occidentalis at different concentrations. The LC₃₀, LC₅₀ and LC₉₀ values of spirotetramat were all much lower than those of imidacloprid after the same time of treatment. Frankliniella occidentalis was more sensitive when treated with spirotetramat than imidacloprid at the concentrations of 500, 250, 125 and 62.5mg/L, and the median lethal concentrations (LC₅₀) were 285.53, 82.24 and 11.19mg/L at 12, 36 and 72 hours after treatment, respectively. Spirotetramat caused 100% mortality of F. occidentalis at 500 and 250 mg/L after 48 hours treatment, and at the lowest concentration 125 mg/L at 72 hours after treatment, respectively. Field experiments carried out elsewhere have confirmed our findings. Together they allow us to conclude that spirotetramat is a promising pesticide for the control F. occidentalis in China.     

Slowing the Spread of Grapevine Leafroll-Associated Viruses in Commercial Vineyards With Insecticide Control of the Vector,Pseudococcus maritimus (Hemiptera: Pseudococcidae)

Vineyards were surveyed for grapevine leafroll-associated viruses and their insect vectors in New York State’s Finger Lakes region in 2006–2008. Grape mealybug, Pseudococcus maritimus (Erhorn) (Hemiptera: Pseudococcidae), European Fruit Lecanium, Parthenolecanium corni (Bouche), and Cottony Maple Scale, Pulvinaria acericola (Walsh and Riley) (Hemiptera: Coccidae) were identified as vector species in this region. An increase in the incidence of Grapevine leafroll-associated virus 1 (GLRaV-1) and GLRaV-3 was observed in 8 of the 20 vineyards surveyed, which implies transmission by these insect vectors. Two of the vineyards for which a temporal increase in disease incidence was documented were then used to evaluate the efficacy of foliar applications of horticultural oil and two classes of insecticides for control of P. maritimus and for slowing virus spread over 2 years of vine protection. Delayed dormant applications of horticultural oil contributed to control of early season crawlers; however, this was not the case for control of summer populations. Applications of acetamiprid and spirotetramat achieved control in summer populations; however, spirotetramat outperformed acetamiprid in percent reduction of treated compared with control vines and in a side-by-side trial. Vines treated with spirotetramat had a lower percentage of new vines testing positive for GLRaV-1 than control vines after 2 years, while no other spray program altered the increase in incidence of GLRaV-1 or -3.     

螺虫乙酯及其代谢物在梨和土壤中的残留及消解动态

为建立梨和土壤中螺虫乙酯及其代谢物螺虫乙酯-烯醇-糖苷 (S-glu)、螺虫乙酯-酮-羟基 (S-keto)、螺虫乙酯-烯醇 (S-enol) 和螺虫乙酯-单羟基 (S-mono) 的残留分析方法，以及明确螺虫乙酯在梨中的残留规律，采用体积分数为1%的乙酸乙腈为提取剂，以N-丙基乙二胺 (PSA) 和无水硫酸镁为分散净化剂的QuEChERS方法，利用超高效液相色谱-串联质谱 (UPLC-MS/MS) 在选择反应监测模式 (SRM) 下检测，外标法定量。结果显示:螺虫乙酯在0.0005~0.1 mg/L范围内，S-glu在0.005~0.5 mg/L范围内，S-keto、S-enol和S-mono在0.0005~0.5 mg/L范围内各化合物的质量浓度与质谱峰面积间均具有良好的线性关系 (R2 ≥ 0.999)；在0.005~0.7 mg/kg添加水平下，螺虫乙酯及其代谢物在梨果中的平均回收率为84%~109%，相对标准偏差 (RSD) 为1.2%~3.3%；在土壤中平均回收率为86%~102%，RSD为1.1%~3.6%。最低检测浓度 (LOQ)为5 μg/kg。该方法检测速度快、灵敏度高、重现性好，适用于梨和土壤中螺虫乙酯及其代谢物残留的快速检测和确证。按推荐剂量进行田间施药，当梨果成熟采收时，螺虫乙酯及其代谢物在梨中的残留量之和在0.023~0.056 mg/kg之间，低于中国规定的最大残留限量标准 (0.7 mg/kg)；在土壤中的残留量在0~0.015 mg/kg之间。螺虫乙酯及其代谢物在梨果和土壤中的消解动态均符合一级反应动力学方程，半衰期分别为为12.4 d和7.1 d。田间残留试验结果表明，螺虫乙酯用于梨树害虫防治是安全的。