孙琦,
范咏梅^,,
赖柯华,
黄伟康
海南大学 环境与植物保护学院, 海口 570228

作者简介: 孙琦(1994-),女,学士,研究方向为植物保护,E-mail:qisun2016@126.com.

通讯作者: 范咏梅,yongmeifan@126.com ;

基金项目: 国家科技支撑计划项目(2012BAK01B05)
海南省教育厅项目(HNJG2014-07)


中图分类号: X171.5

Effects of Dinotefuran on the Embryonic and Larvae Development and Apoptosis in Zebrafish (Danio rerio)

Sun Qi,
Fan Yongmei^,,
Lai Kehua,
Huang Weikang
College of Environment and Plant Protection, Hainan University, Haikou 570228, China

Corresponding author: Fan Yongmei,yongmeifan@126.com ;

CLC number: X171.5

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摘要:第三代新烟碱类农药呋虫胺作为超高效、广谱性杀虫剂在水稻、蔬菜和水果上广泛应用,因其水溶性高,对水生生物的毒性不容忽视。本研究以斑马鱼胚胎为对象,参考OECD标准,在胚胎受精后30 min内,采用静态法染毒,观察其24 h、48 h、72 h、96 h的生长发育情况,根据死亡数计算呋虫胺对斑马鱼96 h-LC₅₀,采用吖啶橙染色(acridine orange fluorescent,AO-F)和原位末端标记法(TUNEL)2种方法,检测其对斑马鱼96 hpf (hour post-fertilization,hpf)幼鱼的细胞凋亡情况。结果表明:呋虫胺对斑马鱼胚胎的96 h-LC₅₀为10.36 g·L^-1(95%置信区间为7.76~12.93 g·L^-1),属于微毒;较高浓度的呋虫胺能使斑马鱼的摆尾数、内心率、孵化率降低,对生长发育有延迟的作用,可导致部分斑马鱼色素褪去,出现心包囊肿、卵黄囊肿和尾部畸形的现象。且随着浓度的升高,在斑马鱼头部、腹部、尾部均有明显的细胞凋亡情况加重,其中以心脏和内耳尤为明显,呈规律的剂量-效应关系。
关键词: 呋虫胺/
斑马鱼/
细胞凋亡/
吖啶橙染色/
TUNEL染色

Abstract:Dinotefuran, as one of the third-generation neonicotinoid insecticide, is highly potent pesticide widely used to control insects on rice, vegetables and fruit field. However, the toxicity of dinotefuran to aquatic organism is still unclear. This study is to explore the effect of dinotefuran on the embryonic and larvae development and apoptosis in zebrafish (Danio rerio). According to OECD guidelines, the 96 h acute toxicity test was conducted using 30 min post-fertilization zebrafish by static system, and the treated zebrafish embryonic and larvae development, were observed in 24 h, 48 h, 72 h and 96 h respectively. The 96 h-LC₅₀ of dinotefuran for adolescent zebrafish was calculated using Karber's method. Then using acridine orange fluorescent (AO-F) and TUNEL staining, the apoptosis of 96 hour post-fertilization (hpf) zebrafish larvae was tested. The results showed that the 96 h-LC₅₀of dinotefuran for zebrafish was 10.36 g·L^-1, with the 95% confidence interval of 7.76-12.93 g·L^-1, which is the slightly toxic according to the pesticide acute toxicity classification standard. Dinotefuran with higher concentration can reduce the tail swing frequency, heart rate and hatchability of D. rerio, delay the growing development, lead to the disappearance of the pigment in D. rerio and cause the pericardiac cyst, vitelline cyst and tail deformity. With the increase of dinotefuran concentration, there were obvious apoptosis observed in the head, abdomen and tail of zebrafish, especially in the heart and inner ear.
Key words:dinotefuran/
zebrafish/
apoptosis/
acridine orange fluorescent/
TUNEL staining.