陕西省榆林地区鸡异刺线虫的流行情况调查及其核糖体ITS区、线粒体cox1基因的序列分析  

Epidemiological survey of Heterakis gallinarum in Yulin area,Shaanxi Province,and sequence analysis of ribosomal ITS gene and mitochondrial cox1 gene

作　　者：郑星星 李中波 尹会方[3,4] ZHENG Xingxing;LI Zhongbo;YIN Huifang(Yulin Vocational and Technical College,Agricultural College,Yulin 719000,China;HuaiHua Vocational and Technical College,College of Animal Science and Technology,Huaihua 418000,China;College of Life Science,Longyan University,Longyan 360012,China;Fujian Provincial University Engineering Research Center for Prevention and Control of Animal-Brone Zoonoses,Longyan 360012,China)

机构地区：[1]榆林职业技术学院农学院,陕西榆林719000 [2]怀化职业技术学院动物科技学院,湖南怀化418000 [3]龙岩学院生命科学学院动物科技学院,福建龙岩360012 [4]动物源性人兽共患病防控福建省高校工程研究中心,福建龙岩360012

出　　处：《黑龙江畜牧兽医》2025年第3期84-92,101,共10页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金区域创新联合基金重点项目(U22A20518);湖南省自然科学基金项目(2022JJ50319);湖南省教育厅项目(21B1072);动物源性人兽共患病防控福建省高校工程研究中心项目(2022K004);怀化市科技局重点项目(2023R2208;2020R2206)。

摘　　要：为了解陕西省榆林地区鸡异刺线虫的流行情况及其核糖体ITS区、线粒体cox1基因序列进化特征,试验于2022年4—10月份采集榆林市榆阳区、横山区、佳县、米脂县、靖边县、定边县和神木县7个区(县)养鸡场的新鲜鸡粪样本536份(笼养鸡和放养鸡粪便各268份),采用饱和盐水漂浮法富集虫卵,并通过形态学检测鸡异刺线虫虫卵,统计不同区(县)笼养鸡和放养鸡粪便鸡异刺线虫阳性样本数量和阳性率;每个区(县)随机挑选出5只粪检呈鸡异刺线虫阳性的放养鸡(编号为YLHG1~YLHG35),取盲肠内容物提取基因组DNA后对鸡异刺线虫核糖体ITS区、线粒体cox1基因进行PCR扩增与测序,分析核糖体ITS区、线粒体cox1基因序列的碱基组成与遗传差异性,并分别基于鸡异刺线虫核糖体ITS区、线粒体cox1基因构建系统发育树以分析榆林地区鸡异刺线虫的系统发育关系。结果表明:检测到的鸡异刺线虫虫卵外观整体呈淡灰褐色、长椭圆形、轮廓清晰,卵壳光滑且分为明显的内外两层,内含单个尚未分裂的胚细胞;536份新鲜鸡粪样本中共有300份样本检出鸡异刺线虫虫卵,总阳性率为55.97%。其中55份来自笼养鸡,阳性率为20.52%;245份来自放养鸡,阳性率为91.42%。榆林市7个区(县)鸡异刺线虫总阳性率由高到低依次为靖边县(64.10%)、定边县(63.41%)、神木县(57.89%)、佳县(55.81%)、米脂县(51.14%)、横山区(50.0%)、榆阳区(46.77%),笼养鸡阳性率由高到低依次为定边县(31.71%)、靖边县(30.77%)、神木县(23.68%)、佳县(18.60%)、米脂县(15.91%)、榆阳区(9.68%)、横山区(9.38%),放养鸡阳性率由高到低依次为靖边县(97.44%)、定边县(95.12%)、佳县(93.02%)、神木县(92.10%)、横山区(90.62%)、米脂县(86.36%)、榆阳区(83.87%)。PCR扩增出鸡异刺线虫的核糖体ITS区、线粒体cox1基因大小分别为969 bp和1325 bp。鸡异刺线虫核糖体ITS区基因序列碱基A、T、G、C含量�In order to understand the prevalence of Heterakis gallinarum and the sequence evolutionary characteristics of ribosomal ITS gene and mitochondrial cox1 gene in Yulin area,Shaanxi Province,in this study,from April to October 2022,536 fresh chicken manure samples were collected from 7 districts and counties of Yuyang District,Hengshan District,Jia County,Mizhi County,Jingbian County,Dingbian County and Shenmu County(268 samples of caged chicken manure and free-range chicken manure each).The saturated saline floatation method was used to enrich the eggs;the eggs of Heterakis gallinarum were detected by morphology.The number and positive rate of positive samples of caged chickens and free-range chicken manure in different regions were counted.5 free-range chickens(numbered YLHG1~YLHG35)were randomly selected from each district(county)and their cecal contents were taken,and genomic DNA was extracted,and PCR amplification and sequencing were carried out for ITS gene and mitochondrial cox1 gene of Heterakis gallinarum.The base composition and genetic differences of ITS gene and cox1 gene sequences were analyzed.Phylogenetic trees were constructed based on the ITS gene and mitochondrial cox1 gene of Heterakis gallinarum to analyze the phylogenetic relationship of Heterakis gallinarum in this region.The results showed that the eggs of the detected Heterakis gallinarum were light gray-brown,oblong and clearly contoured,and the egg shell was smooth and divided into two distinct layers,including a single embryonic cell that had not yet divided.A total of 300 of the 536 fresh chicken manure samples contained eggs of Heterakis gallinarum,with an overall positive rate of 55.97%.Among them,55 were from caged chickens,with a positive rate of 20.52%;245 were from free-range chickens,with a positivity rate of 91.42%.The total positive rate of Heterakis gallinarum in the seven regions of Yulin City was Jingbian County(64.10%),Dingbian County(63.41%),Shenmu County(57.89%),Jia County(55.81%),Mizhi County(51.14%),Hengshan District(50.

关 键 词：鸡异刺线虫 核糖体ITS区基因 线粒体cox1基因 序列特征 基因变异 遗传多样性 系统发育关系 

分 类 号：S852.723[农业科学—基础兽医学] S858.292[农业科学—兽医学]