血凝反应法检测黄毛鼠对抗凝血杀鼠剂抗性的可行性研究

中国媒介生物学及控制杂志 ›› 2013, Vol. 24 ›› Issue (3): 208-210,214.

血凝反应法检测黄毛鼠对抗凝血杀鼠剂抗性的可行性研究

隋晶晶, 高志祥, 姚丹丹, 冯志勇, 颜世祥

广东省农业科学院植物保护研究所, 广东省植物保护新技术重点实验室, 广东广州 510640

出版日期:2013-06-20 发布日期:2013-06-20
通讯作者: 冯志勇,Email: fengzhy@tom.com
作者简介:隋晶晶(1978- ),女,硕士,助理研究员,主要从事鼠类行为生态和鼠害可持续控制技术研究。Email: viola_sui@163.com
基金资助:
“十二五”国家科技支撑计划(2012BAD19B02);广东省自然科学基金(S2011010000895);广东省科技计划项目(2011B031500001)

Study on the feasibility of blood clotting response test for determining resistance to anticoagulant rodenticide in Rattus losea

SUI Jing-jing, GAO Zhi-xiang, YAO Dan-dan, FENG Zhi-yong, YAN Shi-xiang

Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Plant Protection Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, Guangdong Province, China

Online:2013-06-20 Published:2013-06-20
Supported by:
Supported by the "Eleven-Five" National Science and Technology Support Program(No. 2012BAD19B02), Natural Science Foundation of Guangdong Province(No. S2011010000895) and Science and Technology Plan Projects of Guangdong Province (No. 2011B031500001)

摘要/Abstract

摘要：

目的探讨血凝反应法检测黄毛鼠对抗凝血杀鼠剂抗性的可行性。方法以杀鼠灵(warfarin)抗药性区分剂量(10 mg/kg)单次灌胃处理结合致死期食毒抗性检测法(LFP)筛选出黄毛鼠敏感种群和抗性种群,在不同时间段采集试鼠血浆,通过检测试鼠血浆的凝血酶活度(PCA)建立黄毛鼠凝血反应标准曲线,并分析抗性区分剂量处理后抗性个体与敏感个体PCA的变化差异。结果建立了黄毛鼠的凝血反应标准曲线:INR(y)=34.984/x+0.688(x=PCA)(R²=0.992);以10 mg/kg为区分剂量单次灌胃处理后,抗药性黄毛鼠个体的PCA虽有所下降,但可在2～3 d内恢复到正常凝血水平的17%左右;敏感个体的PCA可下降到很低,且不能恢复。结论证实了以血凝反应法检测黄毛鼠对抗凝血剂抗药性的可行性:杀鼠灵10 mg/kg为区分剂量单次灌胃处理4 d后,以PCA=16.5(或INR=4.4)作为阈值来区分黄毛鼠抗药性与敏感性个体,是准确、简便的抗药性判定方法。

关键词: 黄毛鼠, 杀鼠灵, 抗性检测, 血凝反应

Abstract:

Objective To investigate the feasibility of blood clotting response (BCR) test for determining the resistance to anticoagulant rodenticide in Rattus losea. Methods Sensitive and resistant populations of R. losea were screened out by a single gavage of warfarin at a discriminating dose of 10 mg/kg and lethal feeding period test. Plasma was collected from test rats at different time points; the standard curve of BCR was established by measuring the percentage clotting activity (PCA) of plasma, and the difference in PCA between resistant and sensitive individuals treated with a discriminating dose of warfarin was analyzed. Results The standard curve of BCR in R. losea was established as follows: INR(y)=34.984/x+0.688(x=PCA)(R²=0.992). After the single gavage of warfarin at a discriminating dose of 10 mg/kg, the PCA of resistant individuals decreased, but it was restored to 17% of normal level within 2-3 d; the PCA of sensitive individuals decreased dramatically and was not restored. Conclusion This study confirms the feasibility of BCR test for determining the resistance to anticoagulant rodenticide in R. losea. It is accurate and simple to differentiate resistant and sensitive individuals of R. losea by a single gavage of warfarin (10 mg/kg) and using PCA=16.5 (or INR=4.4) as the threshold 4 d later.

Key words: Rattus losea, Warfarin, Determination of resistance, Blood clotting response

中图分类号:

隋晶晶, 高志祥, 姚丹丹, 冯志勇, 颜世祥. 血凝反应法检测黄毛鼠对抗凝血杀鼠剂抗性的可行性研究[J]. 中国媒介生物学及控制杂志, 2013, 24(3): 208-210,214.

SUI Jing-jing, GAO Zhi-xiang, YAO Dan-dan, FENG Zhi-yong, YAN Shi-xiang. Study on the feasibility of blood clotting response test for determining resistance to anticoagulant rodenticide in Rattus losea[J]. Chines Journal of Vector Biology and Control, 2013, 24(3): 208-210,214.

参考文献

[1] 黄秀清,冯志勇,颜世祥,等. 珠江三角洲作物结构变动与害鼠可持续控制技术研究
[J]. 广东农业科学,2004(2):31-34.

[2] WHO. Instruction for determining the susceptible or resistance of rodents to anticoagulant rodenticides
[R]. World Health Organization Technical report:Series no 843,1982:9.

[3] 董天义. 抗凝血灭鼠剂应用研究
[M]. 北京:中国科学技术出版社,2001:1-2.

[4] 冯志勇,姚丹丹,黄立胜,等. 黄毛鼠对第一代抗凝血灭鼠剂的抗药性监测
[J]. 植物保护学报,2007,34(4):420-424.

[5] Wang JS,Feng ZY,Yao DD,et al. Warfarin resistance in Rattus losea in Guangdong province,China
[J]. Pest Biochem Physiol, 2008,91(2):90-95.

[6] MacNicoll AD. The role of altered vitamin K metabolism in anticoagulant resistance in rodents:In Seventeenth Steenbock symposium on current advances in vitamin research(Edited by Sutt JW.)
[M]. New York:Elsevier,1988:407-417.

[7] MacNicoll AD,Gill JE. Revised methodology for a blood clotting response test for identification of warfarin-resistant Norway rat (Rattus norvegicus Berk)
[J]. EPPO Bulletin,1993,23(1): 701-707.

[8] Prescott CV,Buckle AP. Blood-clotting response tests for resistance to diphacinone and chlorophacinone in the Norway rat(Rattus norvegicus Berk)
[J]. Crop Prot,2000,19(5):467-473.

[9] Gill JE,Kerins GM,Langton SD,et al. The development of a blood clotting responses test for discriminating between difenaoum-resistant and susceptible Norway rat(Rattus norvegicus Berk)
[J]. Comp Biochem Physiol,1993,104C(1):29-36.

[10] OEPP/EPPO. Guideline for the evaluation of resistance to plant protection Products:Testing rodents for resistance to anticoagulant rodenticides
[J]. EPPO Bull,1995,25(3):575-593.

[11] Prescott CV. A reappraisal of blood clotting response tests for anticoagulant resistance and a proposal for standardized BCR test method
[S]. RRAC Monograph No.1 Crop International,Brussels, Belgium,2003.

[12] Thijssen HH. Warfarin-based rodenticides:mode of action and mechanism of resistance
[J]. Pestic Sci,1995,43(1):73-78.

[13] 姚丹丹,冯志勇,高志祥,等. 广州市黄毛鼠对抗凝血灭鼠剂的回避行为研究
[M]/全国农业技术推广服务中心. 科学用药,绿色防控. 北京:中国农业出版社,2011:408-412.

[14] 孙毅,梁练,易建荣,等. 黄胸鼠对溴敌隆抗药性的区分剂量测定及对其凝血反应的影响
[J]. 中国媒介生物学及控制杂志,2007, 18(1):4-8.

[1]	姚丹丹, 姜洪雪, 隋晶晶, 冯志勇. 抗凝血杀鼠剂选择压力下黄毛鼠抗药性相关解毒酶的研究[J]. 中国媒介生物学及控制杂志, 2020, 31(6): 657-661.
[2]	姚丹丹, 姜洪雪, 刘福佳, 冯志勇. 广东省江门市黄毛鼠对第一代抗凝血杀鼠剂的抗药性及其与VKORC1基因的相关性研究[J]. 中国媒介生物学及控制杂志, 2019, 30(6): 613-617.
[3]	刘阳, 张韶华, 梁焯南, 朱毅朝, 梁桂. 广东省深圳市2017年褐家鼠对抗凝血杀鼠剂抗药性研究[J]. 中国媒介生物学及控制杂志, 2018, 29(6): 617-620.
[4]	胡群, 马思杰, 裘炯良. 4种DNA条形码在黄毛鼠种类鉴定中的比较[J]. 中国媒介生物学及控制杂志, 2015, 26(3): 286-289.
[5]	姚丹丹, 冯志勇, 隋晶晶, 颜世祥. 黄毛鼠对抗凝血杀鼠剂回避行为的初步研究[J]. 中国媒介生物学及控制杂志, 2013, 24(3): 211-214.
[6]	邓良利, 马林, 孙毅, 时炜, 田汶佳, 何建邯, 廖骏, 肖霖, 刘竹, 张静, 蒋峥. 成都地区褐家鼠种群对抗凝血类杀鼠剂的抗药性水平[J]. 中国媒介生物学及控制杂志, 2012, 23(3): 239-241.
[7]	高志祥, 邱俊荣, 林伟松, 冯志勇, 姚丹丹, 隋晶晶. 湛江市褐家鼠对杀鼠灵的抗药性调查[J]. 中国媒介生物学及控制杂志, 2011, 22(6): 535-537.
[8]	常文英, 郭学林, 宁振东, 王庭林, 邹波, 杨新根, 侯玉, 朱文雅. 山西省娄烦地区北社鼠对杀鼠灵的敏感性研究[J]. 中国媒介生物学及控制杂志, 2011, 22(6): 541-542,549.
[9]	高志祥, 冯志勇, 郭永旺, 姚丹丹, 隋晶晶, 曾繁娟. 新型处方灭鼠诱饵的初步研究[J]. 中国媒介生物学及控制杂志, 2011, 22(5): 436-439.
[10]	高志祥, 邱俊荣, 冯志勇, 姚丹丹, 隋晶晶, 曾繁娟. 雷州市黄胸鼠对杀鼠灵的抗药性调查[J]. 中国媒介生物学及控制杂志, 2011, 22(1): 35-37.
[11]	常文英, 宁振东, 王庭林, 邹波, 杨新根, 郭学林, 侯玉, 朱文雅. 山西省隰县大仓鼠对杀鼠灵的抗药性研究[J]. 中国媒介生物学及控制杂志, 2010, 21(6): 537-538.
[12]	刘大鹏, 张爱军, 吴健, 杨维芳, 褚宏亮, 邵忠贤, 周明浩. 杀鼠灵对黄胸鼠杀灭效果及其凝血酶原时间值测定[J]. 中国媒介生物学及控制杂志, 2010, 21(6): 581-582.
[13]	龙浩宇,王军建,黄志雄,彭亚南. 黄胸鼠对杀鼠灵和溴敌隆抗药性实验室检测报告[J]. 中国媒介生物学及控制杂志, 2010, 21(4): 378-379.
[14]	易建荣1，段金花1，林立丰1，张世炎2，梁练2，阴伟雄1. 广州市斯氏屋顶鼠对第一代抗凝血灭鼠剂抗药性初步研究[J]. 中国媒介生物学及控制杂志, 2009, 20(4): 317-318.
[15]	高志祥1，施大钊2，邱俊荣1，冯志勇1，姚丹丹1，隋晶晶1. 抗凝血类灭鼠剂抗性检测方法的研究概况[J]. 中国媒介生物学及控制杂志, 2009, 20(1): 88-90.