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本文分别用0.02mg/L-0.1mg/L系列浓度的夹竹桃三萜总皂甙水溶液处理钉螺,设无氯清水饲养与浓度为1mg/L氯硝柳胺溶液浸杀对照,研究夹竹桃三萜总皂甙是否为夹竹桃灭螺的化感物质及其作用机理。结果表明:用不同浓度梯度的夹竹桃三萜总皂甙水溶液处理钉螺均有一定的杀灭作用,对试验数据进行概率单位回归分析,表明浸杀处理2、3、4和5d的LC50分别为78.31、30.26、20.50、14.19mg/L,其对应的95%置信区间分别为63.60-108.19、9.49-44.42、2.86-30.90、0.23-22.79mg/L;以不同处理浓度、时间下钉螺的死亡率为因变量,对夹竹桃三萜总皂甙水溶液浸杀灭螺效果进行方差分析,结果显示夹竹桃三萜总皂甙的浓度、处理时间的长短对钉螺的死亡率均有极显著的作用效果,并且药液浓度与处理时间之间的交互效应也极显著。扫描电镜观察结果显示40mg/L浓度处理后钉螺软体表层呈现明显的损伤;透射电镜观察40mg/L浓度处理后钉螺样品发现24h后肝细胞核肿胀,核仁消失,核质稀疏,内质网断裂,线粒体增多;当处理时间延长到48h时,损伤加重,内质网几乎全部囊泡化,细胞核开始破裂,线粒体也破裂;处理24h后钉螺小肠表面的微绒毛排列有些不整齐,有一小部分微绒毛出现脱落,肠周缘细胞出现肿胀,细胞核出现少量小空泡;处理48h时的钉螺小肠表面微绒毛排列明显不整齐,大部分微绒毛脱落,肠周缘细胞肿胀明显,有大量空泡,有时还出现游离核,胞质内的细胞器有大小不等的空泡样变性,细胞器结构已无法辨别,严重变性,胞膜不完整。采用聚丙稀酰胺电泳技术分离经40mg/L夹竹桃三萜总皂甙水溶液处理1、2、3、4、5d后的钉螺样品的酯酶同工酶,处理1-2d后酶的活性比清水对照增强,应激反应明显,而处理达3d时酶的活性开始下降,5d时,酶谱带数及其色泽与用1mg/L氯硝柳胺水溶液处理1d的对照相当。本研究获得化感作用植物夹竹桃灭螺的超微结构证据。
In this paper, snails were treated with aqueous solution of triterpene triterpenoids at a concentration of 0.02mg / L-0.1mg / L, respectively. Chlorine-free water and 1mg / L niclosamide solution were used as control to study the effects of oleander Triterpenoid saponins are the allelochemicals of Oleander snail and its mechanism of action. The results showed that the treatment of Oncomelania snails by triterpene saponin solution with different concentration gradient had a certain killing effect. The regression analysis of the test data showed that the LC50 of 2, 3, 4 and 5 d of the soaking treatment were 78.31,30.26,20.50,14.19mg / L, the corresponding 95% confidence intervals were 63.60-108.19,9.49-44.42,2.86-30.90,0.23-22.79mg / L; at different concentrations, the mortality of snails As the dependent variable, the results of the ANOVA test showed that the concentration of triterpene saponin and the treatment time had an extremely significant effect on the mortality of Oncomelania snails , And the interaction between drug solution concentration and treatment time is also significant. The results of scanning electron microscopy showed that the surface of Oncomelania hupensis showed obvious damage after treatment with 40 mg / L of concentration. The transmission electron microscopy showed that after treatment with 40 mg / L of Oncomelania snail, the nucleus swollen, disappeared nucleolus, sparse nuclei, endoplasmic reticulum rupture, Mitochondria increased; when the treatment time was extended to 48h, the injury increased, the endoplasmic reticulum almost all vesicle, the nucleus began to rupture, mitochondria also rupture; processing 24h after the small snail intestinal microvillus arranged irregularly, a small part of the microvilli Appeared off, intestinal peritumoral cells appear swelling, a small amount of small vacuoles nuclei; processing 48h snail intestinal microvilli arranged significantly irregular, most of the microvilli shedding, intestinal peritumoral cell swelling significantly, a large number of vacuoles, and sometimes also appear Free nuclear, cytoplasmic organelles of varying sizes of vacuolar degeneration, organelle structure has been unable to distinguish, severe degeneration, imperfect membrane. The polyacrylamide gel electrophoresis was used to isolate esterase isozymes from snails fed with 40mg / L Nerium triterpene saponins for 1, 2, 3, 4 and 5 days. The control increased and the stress response was obvious. However, the enzyme activity started to decline after 3 days of treatment. At 5 days, the number of bands and the color of the enzyme were similar to that of 1 mg / L niclosamide aqueous solution for 1 day. In this study, we obtained the ultrastructural evidence of allelopathic plants molluscum.