摘要
基于安全、高效使用杀虫剂溴虫腈防治农业害虫的目的,本文研制了10%溴虫腈水乳剂、微乳剂、悬浮剂和乳油4种剂型;进行了4种剂型的物化特性比较;用气相色谱法检测了溴虫腈不同剂型在桑叶和甘蓝叶片内滞留量和对小菜蛾幼虫的表皮穿透力;比较了溴虫腈不同剂型对桑叶光合特性及叶质的影响;分别用浸叶法和浸虫法测定了溴虫腈不同剂型对家蚕和小菜蛾的毒力;比较了溴虫腈不同剂型处理桑叶后对家蚕生长发育的影响;采用食下毒叶法检测了溴虫腈不同剂型处理桑叶后对家蚕的残留毒性;用气相色谱法检测了喷药后不同时间,溴虫腈不同剂型在桑叶中的降解动态;测定了溴虫腈不同剂型在桑叶上的最终残留量。研究结果如下:
1.通过对溶剂及表面活性剂等的筛选和配伍,确定了10%溴虫腈水乳剂和10%溴虫腈微乳剂的优化配方;采用湿式超微粉碎法,对润湿分散剂、增稠剂、防冻剂等进行筛选,确定了10%溴虫腈悬浮剂的优化配方;对溶剂、乳化剂进行了筛选和配伍,确定了10%溴虫腈乳油的优化配方。研制出了符合国家同类剂型质量标准的10%溴虫腈悬浮剂、10%溴虫腈微乳剂、10%溴虫腈水乳剂和10%溴虫腈乳油4种剂型。
2.溴虫腈4种剂型的物化特性比较显示:溴虫腈4种剂型的表面张力具有比较一致的规律,即稀释倍数越低,浓度越大,表面张力越小;反之,稀释倍数越高,浓度越小,表面张力越大。在相同稀释浓度下,溴虫腈4种剂型的表面张力从大到小顺序依次为10%溴虫腈悬浮剂> 10%溴虫腈水乳剂> 10%溴虫腈乳油> 10%溴虫腈微乳剂。稀释倍数越高,4种剂型稀释液之间的表面张力差距越大;溴虫腈4种剂型在甘蓝叶面的接触角具有相同的规律,即稀释倍数越低,浓度越大,接触角越小。4种剂型在相同稀释浓度下,其稀释液的接触角从大到小顺序依次为10%溴虫腈悬浮剂>10%溴虫腈水乳剂> 10%溴虫腈乳油>10%溴虫腈微乳剂;溴虫腈4种剂型在蜡面上的润湿展布能力依次为10%溴虫腈微乳剂>10%溴虫腈乳油>10%溴虫腈水乳剂>10%溴虫腈悬浮剂,润湿展布面积分别是水的3.08倍、2.60倍、2.36倍和1.38倍;在桑叶上润湿展布面积大小依次为10%溴虫腈微乳剂> 10%溴虫腈水乳剂>10%溴虫腈乳油>10%溴虫腈悬浮剂,润湿展布面积分别是水的2.30倍、2.28倍、1.16倍和1.08倍。在甘蓝叶上润湿展布面积大小依次为10%溴虫腈微乳剂> 10%溴虫腈乳油>10%溴虫腈水乳剂>10%溴虫腈悬浮剂,润湿展布面积分别是水的2.49倍、2.25倍、1.39倍和1.36倍。
3.溴虫腈4种剂型处理桑叶48 h后,在叶片内溴虫腈的滞留量占处理药剂总量的百分率分别为:溴虫腈微乳剂54.6%,溴虫腈乳油47.0%,溴虫腈悬浮剂39.2%,溴虫腈水乳剂37.6%;溴虫腈4种剂型处理甘蓝叶48 h后,在叶片内溴虫腈的滞留量占处理药剂总量的百分率分别为:溴虫腈微乳剂50.2%,溴虫腈乳油44.4%,溴虫腈水乳剂43.2%,溴虫腈悬浮剂34%;溴虫腈4种剂型对小菜蛾表皮穿透能力以溴虫腈微乳剂最高,其次是溴虫腈乳油,再次是溴虫腈悬浮剂,溴虫腈水乳剂的穿透率最低。溴虫腈微乳剂和溴虫腈乳油的穿透率与处理浓度并不表现为倍数关系,而溴虫腈悬浮剂和溴虫腈水乳剂的穿透率与处理浓度的相关性则十分明显。在100 mg·L-1处理浓度下,9h后的穿透率大小依次为:溴虫腈微乳剂88.30%、溴虫腈乳油84.82%、溴虫腈悬浮剂50.76%和溴虫腈水乳剂44.55%。
4.用4种剂型溴虫腈药液喷施桑树后,桑叶的净光合速率、气孔导度、蒸腾速率、蛋白质含量、可溶性糖含量与对照相比均有不同程度的提高,对桑叶净光合速率的影响为微乳剂>悬浮剂>乳油>水乳剂,微乳剂促进桑叶蛋白质和可溶性糖含量的增加也高于其它处理;4种剂型溴虫腈药液喷施后的桑叶脂肪含量与对照相比略有下降,至施药第6d后脂肪含量逐渐恢复到对照水平;各种剂型溴虫腈药液喷施后的桑叶胞间CO2浓度变化没有明显规律。研究结果表明,溴虫腈的4种剂型对桑叶光合特性及叶质均无不利影响,但不同剂型对叶质的影响有一定的差异。
5.用食下毒叶法测得溴虫腈乳油、微乳剂、水乳剂和悬浮剂对2龄期家蚕的LC50值分别为286.23、296.25、313.02和329.94 mg·L-1。用浸虫法测得溴虫腈乳油、微乳剂、水乳剂和悬浮剂对3龄期家蚕的LC50值分别为548.77、492.98、589.52和554.47 mg·L-1。用浸虫法测得乳油、微乳剂、水乳剂和悬浮剂对4龄小菜蛾幼虫的LC50值分别为20.958、12.368、29.402和36.773 mg·L-1。用浸叶法测得乳油、微乳剂、水乳剂和悬浮剂对4龄小菜蛾幼虫的LC50值分别为9.0186、8.2699、20.957和10.815 mg·L-1。采用浸叶法和浸虫法测得的不同剂型溴虫腈对小菜蛾的毒力存在显著差异,浸叶法对小菜蛾幼虫的毒力更高,说明溴虫腈杀虫作用是胃毒作用>触杀作用。
6.100 mg·L-1的溴虫腈悬浮剂、水乳剂、微乳剂、乳油和敌敌畏乳油处理桑树,药后7d起饲喂2龄起蚕,对家蚕幼虫的龄期、眠蚕体重、全茧量(即茧重)、茧层量(茧皮重)、蛹重和化蛹率均没有影响,各处理间均无显著差异。
7.溴虫腈不同剂型处理桑叶后对家蚕的残留毒性测定结果显示,50 mg·L-1的溴虫腈悬浮剂、水乳剂、微乳剂和乳油处理下,均无家蚕中毒与死亡,对家蚕生长发育的影响与对照相比无显著差异;100 mg·L-1溴虫腈不同剂型处理后24 h,家蚕均有死亡。处理后48 h,只有溴虫腈乳油处理的家蚕有死亡,其余处理均无家蚕中毒与死亡,对家蚕生长发育的影响与对照相比无显著差异。处理后72 h,不同剂型处理均无家蚕中毒与死亡;150 mg·L-1溴虫腈不同剂型处理后96 h,只有溴虫腈乳油处理的家蚕有死亡,其余处理均无家蚕中毒与死亡,对家蚕生长发育的影响与对照相比无显著差异;200 mg·L-1和250 mg/L溴虫腈不同剂型处理后120 h均有家蚕中毒与死亡现象,存活个体的生长发育缓慢,体型较对照显著变小。中毒家蚕的症状主要表现为拒食、不活泼、生长发育受阻,中毒较重者身体严重萎缩、卷曲成“S”型,个别中毒较深者吐水死亡。
8.溴虫腈不同剂型在桑叶中的降解动态方程及半衰期分别为:溴虫腈悬浮剂在桑叶中的降解动态方程为C =93.466e-0.1677t (R2 = 0.9238),半衰期为4.1 d;溴虫腈水乳剂在桑叶中的降解动态方程为C = 97.65e-0.234t (R2 = 0.8808),半衰期为3.0 d;溴虫腈微乳剂在桑叶中的降解动态方程为C = 97.675e-0.2261t (R2 = 0.855),半衰期为3.1 d;溴虫腈乳油在桑叶中的降解动态方程为C =124.85e-0.2287t (R2 = 0.8781),半衰期为3.0 d。此检测方法的最低检出浓度为3.97×10-4 mg·kg-1。在施药28 d后,100 mg·L-1溴虫腈处理的桑叶中悬浮剂、水乳剂、微乳剂和乳油残留量分别为:0.019、0.017、0.012和0.016 mg·kg-1。
9.桑园内分别喷施250、200、150、100和50 mg·L-1 4种剂型的溴虫腈,检测到药剂在桑叶的残留毒性期分别为:溴虫腈悬浮剂, >5 d、>5 d、3 d、1 d和0 d;溴虫腈微乳剂,>5 d、>5 d、3 d、1 d和0 d;溴虫腈水乳剂,>5 d、4 d、2d、1 d和0 d;溴虫腈乳油>5 d、>5 d、>5 d、2 d和0 d。
10.10%溴虫腈4种剂型100 mg·L-1喷施桑树后的安全间隔期为施药后5 d,200mg·L-1处理的安全采收期为施药后14 d。10%溴虫腈4种剂型的药剂在桑树上防治害虫或害螨,建议使用浓度为100 mg·L-1。
11.综合溴虫腈4种加工剂型的特性与毒效关系,对桑叶的生理效应和对家蚕的残留毒性等,证明溴虫腈微乳剂是适宜在桑园应用的高效、安全、经济和环保剂型。
In order to exploit chlorfenapyr to control pests safely and efficiently in mulberry fields, the four formulations of chlorfenapyr were designed, compared the physicochemical characteristics of the different formulations, detected the residual amount in the mulberry and cabbage leaves and the penetration rate on the epidermis of diamondback moth of different formulations of chlorfenapyr by GC, compared the effect of the different formulations of chlorfenapyr to the photosynthetic characteristics and the leaf qualities. The toxicity of chlorfenapyr with different formulations to the silkworm and diamondback moth were determined by the leaf and pest dipping methods, compared the effect of chlorfenapyr with different formulations to the growth of silkworm and also determined the residual toxicity of the chlorfenapyr with different formulations to the silkworm, detected the degradation dynamics of chlorfenapyr and the final residual amount of chlorfenapyr in the mulberry leaves.
The results are as follows:
1. The 10% chlorfenapyr water emulsion and the 10% chlorfenapyr microemulsion were designed successfully by screening and combining the organic solvent and the surface activating agent et al. And also the 10% suspension concentrate was designed by screening the wetting dispersant, thickening agent and antifreeze with the wet superfine grinding method. Simultaneously, 10% chlorfenapyr emulsifiable concentrate was designed by screening the organic solvent and the emulsifier. By determination and comparison with the same kinds of the formulations, the four formulations of chlorfenapyr were accord with the standard of the nation.
2. The results of the physicochemical characteristics of the four formulations show that the surface tension of the four formulations is of the same regularity. The lower the dilution, the smaller the surface tension. On the contrary, the higher the dilution, the greater the surface tension. In the same dilution ratio, the sequences of the surface tension of the four formulations are as follows: 10% chlorfenapyr suspension concentrate>10% chlorfenapyr water emulsion>10% chlorfenapyr emulsifiable concentrate>10% chlorfenapyr microemulsion. The larger the dilution ratio, the bigger the distance of the surface tension between the four formulations of chlorfenapyr. The contact angles of the four formulations are of the same regularity, namely, the lower the dilution ratio, the smaller the contact angle. At the same dilution ratio of the four formulations, the sequences of the four formulations are as follows: 10% chlorfenapyr suspension concentrate>10% chlorfenapyr water emulsion>10% chlorfenapyr emulsifiable concentrate>10% chlorfenapyr microemulsion. The sequences of the distribution area of the four formulations on the wax surface are as follows: 10% chlorfenapyr microemulsion>10% chlorfenapyr emulsifiable concentrate>10% chlorfenapyr water emulsion>10% chlorfenapyr suspension concentrate, the distribution areas are 3.08, 2.60, 2.36 and 1.38 times of the water’s. The sequence on the mulberry leaf are as follows: 10% chlorfenapyr microemulsion>10% chlorfenapyr water emulsion>10% chlorfenapyr emulsifiable concentrate>10% chlorfenapyr suspension concentrate.The distribution area of the four formulations are 2.30, 2.28,1.16 and 1.08 times of the water’s on the mulberry leaves. And, the distribution areas of the four formulations on the cabbage leaves are as follows: 10% chlorfenapyr microemulsion>10% chlorfenapyr emulsifiable concentrate>10% chlorfenapyr water emulsion>10% chlorfenapyr suspension concentrate and the distribution area are 2.49, 2.25, 1.39, and 1.36 times of the water’s.
3. After 48h treated with the the four formulations of chlorfenapyr, the residual amount of the pesticides in the mulberry leaves took up 54.6% of the total amount of the chlorfenapyr when treated with the chlorfenapyr microemulsion, and the emulsifiable concentrate, 47.0%, the suspension concentrate, 39.2%, the water emulsion, 37.6%. After 48h treated with the the four formulations of chlorfenapyr, the residual amount of the chlorfenapyr in the cabbage leaves took up 50.2% of the total amont of the chlorfenapyr when treated with the chlorfenapyr microemulsion, and the chlorfenapyr emulsifiable concentrate, 44.4%, the water emulsion, 43.2%, the suspension concentrate, 34%.
4. The results show that the net photosynthetic rate, stomatal conductance, transpiration rate and the protein and soluble sugar contents increase in various degrees treated by 4 formulations of chlorfenapyr in mulberry leaves compared to the control. The effects on the net photosynthetic rate are microemulsion > suspension concentrate > emulsifiable concentrate > water emulsion, and the effects of microemulsion on the protein and soluble sugar contents are most significant. The contents of fat decrease slightly in the prior period after treating compared to the control and recovered to the level of the control after 6 days. The intercellular concentration changes of CO2 do not have obvious rules. The results also indicate that there are no adverse effects on the photosynthetic characteristics and quality of mulberry leaves among the 4 formulations of chlorfenapyr.
5. The 2nd instar larvae of silkworm were treaded with chlorfenapyr emulsifiable concentrate, microemulsion, water emulsion, suspension concentrate using food intake method, the LC50 values are 286.23、296.25、313.02 and 329.94 mg·L-1 respectively. The toxicity of the four different chlorfenapyr formulations to 3nd instar larvae of silkworm were determined by pest dipping method, the LC50 values are 548.77、492.98、589.52 and 554.47 mg·L-1 respectively. The 4nd instar larvaes of diamondback moth were treaded with chlorfenapyr suspension concentrate, water emulsion, microemulsion, emulsifiable concentrate using dipping method, the LC50 values are 20.958、12.368、29.402和36.773 mg·L-1 respectively. The LC50 values of the four formulations to the 4nd instar larvaes of diamondback moth are 9.0186、8.2699、20.957 and 10.815 mg·L-1 respectively. The toxicity of different chlorfenapyr formulations to the 4nd instar larvae of diamondback moth determined by leaf dipping method and pests dipping methods have significant difference, and the toxicity that is determined using leaf dipping method is higher, it indicates that the stomach poisoning of chlorfenapyr is more important than contact killing in the aspects of lethal effect of the diamondback moth.
6. The 2th instar larvae of trimolter and tetramolter were fed with the mulberry leaves which were treated with 100 mg·L-1 chlorfenapyr with the four formulations, the growth and development of the silkworm were observed. The results show that there is no significant effect on the development duration, body weight of molting silkworm, cocoon weight, cocoon shell weight, pupal weight and pupation rate of the silkworm among the four formualatons of the chlorfenapyr.
7. The residual toxicity of chlorfenapyr with different formulations was determined, the results show that the chlorfenapyr with four formulatons at 50mg·L-1 are not harmful to the silkworm all the time and has no significant difference compared with the control. While, when the concentration was increased to 100 mg·L-1, 24h after treated with the four formulations, the silkworms are poisoning and died occasionally. 24h after treated the pesticide, only the chlorfenapyr EC has lethal effect to the silkworm and the other formulations have no effect on the development of the silkworm compared with the control, and there is no poisoning silkworms after 72h treated with the four formulations. When treated with 150 mg·L-1, only chlorfenapyr EC can result in the death of the silkworm after 96h, the other formulations has no effect on the growth of the silkworms and have no significant difference compared with the control. When the chlorfenapyr concentration increased to 200 and 250 mg·L-1, the high concentration can result in the poisoning and death of the silkworms, and the survival is smaller and grow slowly and has significant difference compared with the control.The main poisoning symptoms of the silkworms are antifeedant, inanimation, and hindered growth and development, some serious poisoning silkworm curled into a "S" type, and the individual who poisoned deeper were died. The penetration ability of the chlorfenapyr microemulsion is the most on the epidermis of diamondback moth, the next is the chlorfenapyr suspension concentrate, and the least one is the chlorfenapyr water emulsion. The penetration rate of the chlorfenapyr microemulsion and emulsifiable concentrate is not correlated with the pesticide concentration; while the penetration ability of the chlorfenapyr suspension concentrate and the water emulsion have high correlation with the concentration. When treated with 100 mg·L-1 chlorfenapyr 9 h, the sequences of the penetration ability are as follows: the penetration rate of chlorfenapyr microemulsion is 88.30%, and the chlorfenapyr emulsifiable concentrate, 84.32%, the chlorfenapyr suspension concentrate, 50.76%, the chlorfenapyr water emulsion, 44.55%.
8. The degradation kinetics equation and the half life time of different chlorfenapyr formulations in mulberry leaves are as follows: the degradation kinetics equation of chlorfenapyr SC is C = 93.466e-0.1677t (R2 = 0.9238),the half life time is 4.1d. The degradation kinetics equation of chlorfenapyr EW is C = 97.65e-0.234t (R2 = 0.8808), the half life time is 3.0 d, the degradation kinetics equation of chlorfenapyr ME is C = 97.675e-0.2261t (R2 = 0.855), the half life time is 3.1 d. The degradation kinetics equation of chlorfenapyr EC is C =124.85e-0.2287t (R2 = 0.8781), the half life time is 3.0 d. The minimum detectable concentration of this method is 3.97×10-4 mg·kg-1. The mulberry leaves are treated by four different chlorfenapyr formulations at a concentrate of 100 mg·L-1, and the residual levels are detected after 28 d, the results are as follows: the residual levels of chlorfenapyr SC are 0.019 mg·kg-1, the residual levels of chlorfenapyr EW are 0.017 mg·kg-1, the residual levels of chlorfenapyr ME are 0.012 mg·kg-1, the residual levels of chlorfenapyr EC are 0.016 mg·kg-1.
9. After the mulberry leaves were treated by four chlorfenapyr formulations at concentraton of 250, 200, 150, 100 and 50 mg·L-1, and the period of residual toxicity is detected. The results are as follows: chlorfenapyr SC, > 5 d, > 5 d, 3 d, 1 d and 0 d, chlorfenapyr ME, > 5 d, > 5 d, 3 d, 1 d and 0 d, chlorfenapyr EW, > 5 d, 4 d, 2d, 1 d and 0 d, chlorfenapyr EC> 5 d, > 5 d, > 5 d, 2 d and 0 d.
10. The safety interval of four kinds of 10% chlorfenapyr formulations is 5 days after spraying the chlorfenapyr at 100 mg·L-1, and the safety interval of four kinds of 10% chlorfenapyr formulations is 14 days at 200 mg·L-1. It is recommended to use four kinds of 10% chlorfenapyr formulations to prevent and control the mulberry pests and pest mites at 100 mg·L-1.
11. By the comprehensive analysis of the physicochemical characteristics, the poisonous effect , the physiological effect to the mulberry leaves and the residual toxicity of the four formulations of chlorfenapyr, which indicates that chlorfenapyr microemulsion is of high efficiency to pests, high safety to silkworm and could be utilized as the economical and environmental friendly formulation in the mulberry fields.
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