【作者】 阮风雷；

【导师】 呼和巴特尔；

【作者基本信息】 内蒙古农业大学 ， 预防兽医学， 2007， 硕士

【摘要】 本课题旨在建立有效控制牛皮蝇蛆病在内蒙古地区的流行的科学防治时机及免疫学诊断方法,本研究在牛皮蝇成蝇季节动态调查和幼虫移行规律研究的基础上,确定了内蒙古地区牛皮蝇蛆病的最佳防治时机;为了实现早期诊断和把握防治时机,分别以虫体粗蛋白和重组蛋白为抗原,建立了牛皮蝇蛆病的ELISA诊断方法。防治时机的研究:内蒙古地区牛皮蝇蛆病为纹皮蝇和牛皮蝇混合感染,两者比例为4.1:1,纹皮蝇为优势蝇种。在内蒙古西部呼和浩特市周围农牧交错区纹皮蝇Ⅰ期幼虫在8月下旬开始进入牛食道部,在9月中旬～10月上旬寄生数量达到高峰,12月初纹皮蝇幼虫完全离开食道向背部移行,在牛背部发育为Ⅲ期幼虫,形成“瘤包”。纹皮蝇Ⅲ期幼虫在4月上旬落地,到5月底～6月初成蝇开始出现,6月末成蝇基本消失。牛皮蝇的出没要比纹皮蝇晚15天左右;在东部北方牧区,皮蝇的活动规律与西部基本相似,只是出没时间较西部晚约半个月。根据牛皮蝇的季节动态和幼虫移行规律,确定了内蒙古西部农牧交错区牛皮蝇蛆病的主要防治时机为7月中旬～8月中旬,辅助防治时机为1月中旬～2月中旬;在东部牧区牛皮蝇蛆病的主要防治时机为8月初～9月初,辅助防治时机为2月初～3月初。诊断方法的建立:分别以虫体粗蛋白和重组rHC蛋白为检测抗原建立了牛皮蝇蛆病的ELISA诊断方法。两种抗原的包被浓度分别为26.64μg/ml和15.63μg/ml,均具有较好的特异性和敏感性,虫体抗原的特异性和敏感性分别为100%和98.36%,重组抗原的特异性和敏感性分别为95.8%和100%。用建立的两种ELISA诊断方法检测采自内蒙古地区的233份田间血清,结果显示两种抗原ELISA检测方法间差异不显著(p>0.05)。在次年以传统诊断方法进行复检,两种诊断方法均有很好的阳性符合率(分别为88.46%和93.75%),说明两种抗原建立的ELISA诊断方法均可用于牛皮蝇蛆病的诊断。更多还原

【Abstract】 In order to control the prevalence of hypodermosis in Inner Mongolia,the best preventive and control time and diagnostic method were established.We have investigated the seasonal fluctuation of warble flies and the rule of larvae migration.According to the investigative results, we established the best preventive and control time of Hypodermosis in the eastern and western area in Inner Mongolia. In order to take early diagnosis and hold preventive and control time,a specific serological ELISA method was developed with natural protein and recombinant protein as detective antigen for hypodermosis.The study on the best preventive and control time: there were two species of Hypoderma spp. ,including H.lineatum and H.bovis in Inner Mongolia,and H.lineatum is the local superior species, the ratio of the two species is 4.1:1. In the western Inner Mongolia ,the first instar larvae of hypoderma spp. were discovered in esophagus in the last ten days of August, the highest infection rate occured in September and October,the larvae disappeared from esophagus at the beginning of December and migrated to back of cattle and transformed into the third instar larvae.The third instar larvae of H.lineatum departed from cattle back in the first ten days of April, the warble flies appeared between the last ten days of May and the first ten days of June,and disappeared at the end of June .Compared with H.linertum, the time of H.bovis delayed about fifteen days.The seasonal fluctuation of hypodera spp. is similar in the eastern and the western Inner Mongolia,the difference of the two areas was the appear and disappear time of Hypoderma spp.,and the eastern lated about half a month.According to the investigative results, we established the best preventive and control time of Hypodermosis in the eastern and the western Inner Mongolia. In the western,the main preventive and control time was from the middle ten days of July to the middle ten days of August,and the assistant preventive and control time was from the middle ten days of January to the middle ten days of February .In the eastern, the main preventive and control time was from the beginning of August to the beginning of September,and the assistant preventive and control time was between next February and March.Development of diagnostic method:A specific serological ELISA method was developed with natural protein and recombinant protein as detective antigen for hypodermosis.We found that the best working concentration of the two antigen were 26.64μg/ml and 15.63μg/ml.The results showed that: the two developed ELISA had higher sensitivity and specification for hypodermosis,natural protein antigen ,98.36% and 100% respectively,recombinant protein antigen,100% and 95.8% respectively.we detected 233 blood sera of cattle which collected from in the western and the eastern Inner Mongolia,the results showed that: there was no conspicuous differences between the two antigen (p<0.05).According to the detection with the traditional method next year,we found that the two methods have better positive coincidence ratio ( 88.46% (46/52) and 93.75%(45/48)),respectively.Therefore,the two methods could be used to diagnose hypodermosis.更多还原