Zn2+、Cu2+在黑水虻血淋巴累积及对血细胞影响的研究

Black soldier fly (Hermtia illucens L.) is an important resource insect spread worldwidely. Their larvae feed on animal manures, which can reduce the amount and stink of animal manures, inhibit breeding of housefly effectively, and decrease the number of bacteria in manures; The byproducts (larvae, prepupae and pupae) of black soldier fly are excellent materials that can be used in food, forage and drug industries.Therefore, using black soldier fly larvae to decontaminate animal excreta is a new way of resolving problems of serious animal manure pollution, and shortage of resource in our country. But, non-rational use of additives in feeds has caused excess and accumulation of heavy metals in animal feces. Our previous study results show that excessive Zn2+ and Cu2+ can accumulate in black soldier fly body and have some effects on thir growth and development. So, it's needed to know whether long-erm heavy metal stress on black soldier fly affect its ability of making excreta innocuous and if it will decrease the values of its byproducts. In this project, it is planned to extend the stress time of Zn2+ and Cu2+, and intensively study their accumulation in black soldier fly's hemolymph and the effects to hemocytes.We hope to establish a theoretical basis for manure harmless management system using black soldier fly, and to provide valuable experimental data to food, forage and drug industries.

黑水虻是全球广泛分布的重要资源昆虫，幼虫取食粪便后可以减少粪便堆积及臭气，有效控制家蝇滋生，减少粪便中细菌数量；其副产品（幼虫、预蛹、蛹）又是食品、饲料及医药研发的优良材料。因此，利用黑水虻幼虫进行粪便无害化处理是解决我国日益严重的畜禽粪便污染及资源短缺问题的一条新途径。由于当今禽畜饲料添加剂的不合理使用，使粪便中重金属含量严重超标并大量累积。本项目前期研究已表明，Zn2+、Cu2+可以在黑水虻体内累积并对其生长发育产生一定影响。黑水虻如果长期处于重金属胁迫下，是否会影响其无害化处理粪便能力？是否会对其副产品的研发及应用价值产生影响？这是非常值得进一步深入研究的问题。本项目拟延长Zn2+、Cu2+胁迫时间，深入研究其在黑水虻血淋巴内累积及对血细胞的全面影响。期待通过该研究为黑水虻无害化处理粪便技术的有效实施提供有力的理论依据；为其副产品在饲料、食品及医药研发中的应用提供有价值的实验数据。

黑水虻是呈世界分布的重要资源昆虫，其幼虫被广泛用于畜禽粪便的无害化处理。但是，当今畜禽饲料添加剂的不合理使用，使畜禽粪便中Cu2+、Zn2+含量严重超标，超标的Cu2+、Zn2+可在黑水虻体内累积。黑水虻幼虫如果长期在Cu2+、Zn2+胁迫下会产生怎样影响，是黑水虻幼虫无害化处理粪便技术推广时值得深入研究的问题。本项目不仅研究了Cu2+、Zn2+在第1、3、5代黑水虻幼虫血淋巴内累积，还研究了Cu2+、Zn2+连续5代胁迫对黑水虻幼虫血细胞影响。结果表明， Cu2+、Zn2+ 可在黑水虻幼虫血淋巴内累积且随胁迫浓度及胁迫世代增加含量增加；Cu2+、Zn2+胁迫对血细胞超微结构及凋亡形态产生影响，当处理浓度达到600mg/kg时各个世代均出现细胞核形态不规则，核内染色质凝聚，核仁消失，胞质浓缩，细胞质内出现大量空囊泡，当处理浓度达到1200mg/kg时细胞膜伸出伪足，出现内陷、破裂，有内容物外泄，细胞核裂解；Cu2+、Zn2+连续胁迫亦对幼虫血淋巴中能量物质及酶活产生影响，1200mg/kg处理浓度胁迫可显著抑制能量物质及酶活的产生，且随胁迫世代增加，影响增加；Cu2+、Zn2+连续胁迫不仅对血细胞凋亡产生影响，还导致DNA损伤，长期胁迫还可使幼虫抗菌肽含量降低。. 另外，为了本项目的顺利进行我们还研究了含水量对黑水虻生长发育影响、黑水虻抗菌肽提取条件优化及纯化后抗菌肽对CNE2细胞影响。结果表明，饲料含水量为30%、50%时显著影响黑水虻生长发育；抗菌肽H3可显著抑制CNE2细胞体外增殖，抑制率达40.56±6.80%，160 μg/ml处理浓度H3不仅使CNE2细胞发生显著凋亡，还可显著降低CNE2细胞的迁移率，使其端粒酶活性和hTERT蛋白表达量明显下降。. 本研究将为黑水虻无害化处理粪便技术的实施提供可应用的实验数据，并为其副产品的研发提供有价值的理论依据。