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1.
A sex determination method using DNA extracted from feces has been developed for sika deer (Cervus nippon). We determined a partial sequence of the amelogenin gene of sika deer, which exists on both X and Y chromosomes with a deletion region on the Y chromosome. Based on the sexually dimorphic sequences, we designed a pair of primers which could amplify DNA fragments the lengths of which are different between males and females. PCR products were detected in 34 out of 37 fecal samples collected from captured deer and the sexes estimated by the present method were perfectly matched with the actual sexes.  相似文献   

2.
为了探讨吉林省圈养梅花鹿的遗传分化情况,利用DNA直接测序技术测定其线粒体细胞色素mt DNA Cytb基因全序列,结合Gen Bank收录的其他梅花鹿亚种同源序列,利用生物学软件进行分析。结果表明,15头样本mt DNA Cytb基因序列中,共检测到核苷酸多态位点38个,吉林省圈养梅花鹿种群的遗传多样性水平较低,与俄罗斯、韩国、朝鲜亚种亲缘关系较近,与四川亚种亲缘关系较远。  相似文献   

3.
The aim of the present study was to determine the whole nucleotide sequence of the open reading frame of the sex‐determining region Y (SRY‐ORF) in wild sika deer. The SRY gene of wild sika deer was obtained by polymerase chain reaction (PCR) with DNA from blood samples. The whole nucleotide sequence of the SRY‐ORF in wild sika deer consisted of 687 bp and encoded 229 deduced amino acids. In comparison with the bovine SRY gene, the percentage of nucleotide sequence homology was 91.0% in the overall ORF, and those of the N‐terminal, high mobility group (HMG) box, and C‐terminal regions within ORF were 88.9%, 96.2% and 87.9%, respectively. The nucleotide sequences of sika deer SRY‐ORF characterized in the present study can be used for phylogenetic analysis or sexing in wild sika deer.  相似文献   

4.
The browsing habits of sika deer (Cervus nippon) in Japan have caused serious ecological problems. Appropriate management of sika deer populations requires understanding the different genetic structures of local populations. In the present study, we used 10 microsatellite polymorphisms to explore the genetic structures of sika deer populations (162 individuals) living in the Kanto region. The expected heterozygosity of the Tanzawa mountain range population (Group I) was lower than that of the populations in the Kanto mountain areas (Group II). Our results suggest that moderate gene flow has occurred between the sika deer populations in the Kanto mountain areas (Group II), but not to or from the Tanzawa mountain range population (Group I). Also, genetic structure analysis showed that the Tanzawa population was separated from the other populations. This is probably attributable to a genetic bottleneck that developed in the Tanzawa sika deer population in the 1950s. However, we found that the Tanzawa population has since recovered from the bottleneck situation and now exhibits good genetic diversity. Our results show that it is essential to periodically evaluate the genetic structures of deer populations to develop conservation strategies appropriate to the specific structures of individual populations at any given time.  相似文献   

5.
试验旨在对基于基因分型测序(genotyping by sequencing,GBS)技术筛选出的马鹿特异性SNPs位点的准确性进行验证,为梅花鹿、马鹿及其杂交后代的鉴别提供可靠的分子遗传标记。随机选取30个马鹿特异性SNPs位点,根据SNPs位点前后各200 bp的序列,利用Primer Premier 6.0软件设计特异性引物,以随机选取的验证样本DNA作为模板进行PCR扩增,并进行Sanger测序,对测序结果利用BioEdit软件进行峰图的观察,利用Mega 6.0软件对测序得到的序列进行比对分析并观察每个特异性SNP位点在不同验证群体中的基因型,对每一个马鹿特异性位点的峰图和比对信息进行统计分析。结果表明,30个马鹿特异性位点中有28个和前期研究结果一致,其中1个SNP位点(SNP3)中G等位基因在梅花鹿中的基因频率为0.05,而G等位基因在马鹿中的基因频率为1,G等位基因在马鹿个体中的基因频率比在梅花鹿个体中高,同时,利用SPSS 22.0进行统计分析发现,该位点在马鹿和梅花鹿中基因型分布表现出显著性差异(P<0.05);另外1个SNP位点(SNP5)中T等位基因在梅花鹿的基因频率为0.15,而在马鹿中的基因频率为1,且这个SNP位点在梅花鹿和马鹿中基因型分布差异显著(P<0.05),所以这2个位点仍然可以作为马鹿的特异性SNPs位点。研究结果说明了GBS测序筛选出的马鹿特异性SNPs可以作为鉴定的分子标记,对梅花鹿、马鹿及其杂交后代的鉴别奠定了理论基础。  相似文献   

6.
The feasibility of fecal steroid analysis for pregnancy diagnosis and sex determination were tested in sika deer (Cervus nippon). Feces were collected from captive sika deer in June (non-breeding season and late-pregnancy period) and October (breeding season), and also from the rectum of 24 female sika deer (19 pregnant and 5 non-pregnant females) shot as part of programs for population control in February and March (mid-pregnancy period). In mid- and late-pregnancy periods, fecal progesterone concentrations were significantly higher in pregnant female than in male and non-pregnant female deer. In October, fecal testosterone concentrations were higher in adult male deer, and no difference was found between young males and females. These results suggest that fecal steroid analysis would be a useful means for estimating pregnancy status and for detecting adult male among wild deer.  相似文献   

7.
The 18S rRNA gene and the piroplasm major immunodominant protein gene (p33/34) of Theileria from various subspecies of sika deer in 8 different locations of Japan were analyzed. The similarity between 633 bp partial sequences of the 18S rRNA gene among various subspecies of sika deer was found to be between 99.7% and 100%. While the percent identities of the 412 bp partial p33/34 gene sequence and deduced amino acid sequences between Theileria of sika deer from Yamaguchi Prefecture and those found in deer from other Prefectures, were comparatively low, 68.7% to 70.1% and 64.1% to 70.0% respectively. These findings suggest that there are at least two genetically distinct strains of Theileria of sika deer in Japan.  相似文献   

8.
吉林、黑龙江两省梅花鹿布氏杆菌病的血清学调查   总被引:1,自引:0,他引:1  
应用血清学方法,对我国吉林、黑龙江两省梅花鹿成年鹿群未注射布氏杆菌病疫苗的874份血清进行了血清抗体检测,其抗体阳性率分别为8.87%、2 6.39%。这一数据表明,吉林省和黑龙江省的鹿群中存在着布氏杆菌病的感染。同时只对不同性别的梅花鹿进行了血清抗体检测,结果表明,梅花鹿公鹿和母鹿对布氏杆菌均易感。此研究为今后有效的预防和治疗布氏杆菌病提供了临床理论依据  相似文献   

9.
2018年3月17日,贵州省某梅花鹿场的梅花鹿(Cervus nippon)出现关节肿大、下颌肿大、睾丸单侧肿胀,还伴有严重的跛行等症状。采集4份该发病梅花鹿的血样送检。检测结果表明,通过虎红平板凝集试验2份血样检测出了布氏杆菌感染,4份血样经RT-PCR均扩增出了JEV NS1部分基因的特异性条带,PCR产物测序结果分析显示,4份梅花鹿血样的JEV NS1基因(GZ-015、GZ-019、GZ-022、GZ-039)间的核苷酸同源性分别为99. 9%、99. 3%、99. 4%,与强毒株SA-14、国内外的疫苗株及四川、吉林等地的JEV分离株之间核苷酸同源性为99%以上,GZ-015、GZ-019、GZ-022、GZ-039与强毒株SA-14、弱毒株、国内外的疫苗株及四川、吉林等地的JEV分离株处于同一个较大的遗传进化分支。  相似文献   

10.
We examined the antimicrobial susceptibility of 848 Escherichia coli isolates from 237 feces samples of wild sika deer (Cervus nippon) captured between 2016 and 2019 in 39 of the 47 prefectures of Japan. Five of the 237 wild sika deer (2.1%) carried E. coli with resistance to at least one antimicrobial, and all the resistant isolates showed resistance to tetracycline. The resistant isolates contained antimicrobial resistance genes that were similar to those in E. coli derived from humans and farm animals. Although wild sika deer are not currently likely to be a source for the transmission of antimicrobial resistance in Japan, they can potentially mediate antimicrobial resistance spread by coming into contact with humans, animals, and their surroundings.  相似文献   

11.
In the present study, we tried to detect protozoan blood parasites from the liver or blood of 141 Japanese sika deer (Cervus nippon) in Iwate Prefecture of Japan by polymerase chain reaction. Approximately 500-bp amplicons were obtained in 76 (53.9%) of 141 samples by amplification for V4 hyper-variable regions of the 18S rRNA gene, and the amplicons were considered to be DNA of Theileria species. The complete nucleotide sequences (1701-bp) of the 18S rRNA gene were determined in 25 samples and were divided into 8 genotypes that were phylogenetically separated into two distinct lineages showing a monophyletic relation. The two lineages of Theileria were detected in different rates (12 and 88%) from sika deer in Iwate Prefecture.  相似文献   

12.
本试验通过研究吉林梅花鹿不同生长阶段血液生理生化指标的差异情况,旨在为梅花鹿的健康检测、疾病防治、育种等提供依据和参考。选取吉林省左家、双阳、东丰地区哺乳仔鹿93头(公37头,母56头)、育成鹿135头(公40头,母95头)、成年鹿130头(公42头,母88头),共358头,进行13项血液生理指标、19项生化指标检测。结果显示:哺乳公鹿与哺乳母鹿各项指标总体上差异不大,仅红细胞系统指标、血糖、肌酸激酶具有显著性差异(P<0.05);红细胞压积、红细胞平均体积、平均血红蛋白含量、平均血红蛋白浓度、碱性磷酸酶、乳酸脱氢酶在不同性别的育成鹿与成年鹿中差异极显著(P<0.01)。不同生长阶段公鹿天门冬氨酸氨基转移酶、前白蛋白、总胆汁酸、血糖等差异显著(P<0.05),红细胞系统、肌酸激酶、钙、无机磷、肌酐等差异极显著(P<0.01),白蛋白、淀粉酶差异不显著(P>0.05);不同生长阶段的母鹿淋巴细胞总数、红细胞总数、中性粒细胞总数差异显著(P<0.05),白细胞总数、红细胞系统指标、天门冬氨酸氨基转移酶、丙氨酸氨基转移酶、肌酸激酶、钙、无机磷等差异极显著(P<0.01),谷氨酰转肽酶、总胆汁酸、白蛋白差异不显著(P>0.05)。以上结果说明,不同生长阶段、不同性别的梅花鹿血液生理生化指标存在较大变异,部分指标的差异可能与梅花鹿生茸、性别等生物学特性有关。对上述生理生化指标的测定,获得了不同性别的梅花鹿在不同生长阶段生理生化指标的参考范围,可为梅花鹿选种育种、疾病诊断、资源开发利用提供一定的参考,也可为野生东北梅花鹿资源保护和健康评估提供依据。  相似文献   

13.
Surveillance of chronic wasting disease (CWD) was conducted by performing Western blot analysis of tissue samples from 136 sika deer (Cervus nippon) killed by hunters in the Tokachi district of Hokkaido Island. No prion protein (PrPSc) associated with CWD was detected in any of the samples. To assess amino acid polymorphisms of the sika deer PrP gene, nucleotide sequencing of the PrP gene was performed. The only amino acid polymorphisms detected were 3 silent mutations at nucleotide positions 63, 225 and 408. These results suggest that sika deer in the Tokachi district are genetically homogeneous, and are not infected with CWD.  相似文献   

14.
应用Sry-PCR扩增鉴定狍(Capreolus capreolus)的性别   总被引:2,自引:0,他引:2  
根据人的SRY基因核心序列设计合成1对引物C1、C2,应用PCR技术对野生狍(Capreolus capreolus)Sry基因(哺乳动物Y染色体DNA雄性特异区)进行扩增.结果在野生狍雄性样本扩增出1条带(221bp).而在雌性样本未见扩增带.显示了Sry基因的性别特异性。应用这1对引物对42个未知性别狍肌肉组织样本进行了性别鉴定.结果雄性22个,雌性20个。对Sry-PCR产物克隆测序,得到185bp的Sry基因部分核苷酸序列。本试验为狍种群性别比率及其种群动态变化机制研究提供了资料。  相似文献   

15.
Seven sika deer (Cervus nippon) in a herd of 30 deer in a park died. Upon examination of three dead deer, Salmonella Typhimurium was isolated from the organs and intestinal contents. Histopathological examination revealed catarrhal enteritis and focal necroses in the liver. Immunohistochemically, Salmonella antigen of O4 was detected in the enteric lesions. The case was diagnosed as S. Typhimurium infection in the sika deer. Because of the importance of Salmonella in public health, fecal and soil samples were continuously collected from the paddock. However, no Salmonella was isolated from any samples collected after medication of the deer and thorough disinfection of the immediate environment.  相似文献   

16.
采用血清学方法,对沈阳地区的梅花鹿群未注射布氏杆菌病疫苗的541份血清进行了检测,阳性率为26.62%。结果表明鹿群中仍然存在布氏杆菌病。  相似文献   

17.
旨在利用基因分型测序(genotyping by sequencing,GBS)技术对梅花鹿、马鹿及其杂交后代(F1、F2)基因组的SNP特征进行分析。本试验采用GBS技术对梅花鹿(63个)、马鹿(12个)及其杂交后代(F1代112个,F2代38个,未知类型个体1个)共226个个体的血液基因组DNA进行测序,并利用本实验室前期110只梅花鹿、197只马鹿和1只F1代杂交鹿的测序数据,以梅花鹿全基因组为参考序列进行比对分析。结果,226个个体共产生Clean data 322.683 Gb,平均每个样品1 427.802 Mb;将所有样本作为一个群体检测SNP变异,共检测出SNP位点23 943 582个,质控过滤后得到SNP位点31 630个。对31 630个SNPs使用最大似然(maximum likelihood,ML)法构建的分子进化树显示,梅花鹿、马鹿、F1及F2代区分明显。对梅花鹿和马鹿的SNPs进行比对分析,筛选出可用于鉴别马鹿、梅花鹿、F1、F2的物种特异SNP位点1 032个(马鹿特异SNP位点474个,梅花鹿特异SNP位点558个),计算结果显示,F1代个体包含马鹿特异SNPs的比例主要在40%~60%之间,F2代个体含马鹿特异SNPs的比例主要在10%~30%之间,马鹿个体中不含梅花鹿的特异SNPs,梅花鹿中55.49%的个体不含马鹿特异SNPs,17.34%的个体含马鹿特异SNPs的比例低于1%,13.29%的个体含马鹿特异SNPs的比例在1%~10%之间,其余个体含马鹿特异SNPs的比例为10%~20%(其中有一个个体含马鹿特异SNPs的比例为33.3%)。该研究为花马杂交鹿后代的鉴定提供了可靠标记,并定量估计了F1和F2代个体含马鹿特异SNPs的比例,马鹿个体中不含梅花鹿的特异SNPs,这对梅花鹿、马鹿及其杂交后代(F1、F2)的鉴别具有重要意义。  相似文献   

18.
试验旨在研究梅花鹿SRY基因的遗传多样性及其父系类型。选取东北亚种、北海道亚种、本州亚种、指名亚种、屋久岛亚种5个亚种的144个个体,利用试剂盒和传统酚/仿(1∶1)法进行DNA的提取,采用PCR扩增和直接测序法分析梅花鹿的核苷酸多样性(Pi)、单倍型多样性(Hd)、遗传距离及系统进化关系。结果显示,试验所获序列长度为1 613bp,在第47、62、602、1 148、1 569、1 604bp处共检测到6个SNPs多态位点,占核苷酸总数的0.3%,碱基替换以碱基转换为主。通过6个SNPs多态位点确立了6种单倍型:Hap-1、Hap-2、Hap-3、Hap-4、Hap-5和Hap-6,其中Hap-4、Hap-5和Hap-6为新发现单倍型。遗传多样性由高至低依次为:指名亚种、屋久岛亚种、东北亚种、本州亚种、北海道亚种。各亚种间遗传距离最小为北海道亚种与本州亚种的距离(0.000056),最大为东北亚种与指名亚种的距离(0.001733)。基于单倍型利用邻接法构建系统进化树,结果显示,与日本梅花鹿相比,东北亚种与马鹿关系更近,东北亚种存在两大分支,日本梅花鹿各亚种间无明显分支,其他单倍型均是由Hap-3进化而来,单倍型最小跨度网络图与系统进化树一致。结果表明,东北梅花鹿存在两大父系类型,日本梅花鹿存在一个父系类型,单倍型Hap-3是日本梅花鹿的原始单倍型。  相似文献   

19.
【目的】本研究旨在探索梅花鹿甲硫氨酸亚砜还原酶B3(methionine sulfoxide reductase B3,MSRB3)基因多态性及其与茸重性状的关联性。【方法】选取高、低产茸量的梅花鹿各8只,应用DNA直接测序法检测基因变异位点。采用MassARRAY~? SNP分型技术对314头24月龄梅花鹿MSRB3基因进行基因分型,并结合梅花鹿茸重性状数据进行了关联分析和单倍型分析。【结果】在梅花鹿MSRB3基因中共发现6个SNPs位点,其中2个SNPs位点位于外显子区域,且突变均未引起氨基酸改变,属于同义突变,其余4个SNPs位点均存在于内含子区域。分型结果显示4个样本未分型成功,其余310个样本进行后续分析。各位点观测杂合度和期望杂合度基本一致,g.44455582 T>C、g.44455759 C>T、g.44414424 T>C、g.44350306 T>C及g.44340836 G>A等5个位点的杂合度较高,均属于中度多态(0.25C位点杂合度较低,属于低度多态(PIC<0...  相似文献   

20.
There has been no report on Chronic wasting disease (CWD) cases in Japan to date; however, there is concern about the geographic spread of CWD. To clarify the CWD status in Japan, we conducted CWD monitoring using real-time quaking-induced conversion (RT-QuIC) assay which can detect the low level of CWD prions. A total of 690 obex samples collected from sika deer and Reeves’s muntjac in Hokkaido and Honshu was tested for CWD prions. No CWD-positive cases were found, suggesting that CWD is nonexistent in Japan. Our results also indicate that RT-QuIC assay is useful for continuous monitoring of CWD. Furthermore, nucleotide sequence analysis of the PrP gene revealed sika deer in Japan harbor CWD susceptible allele.  相似文献   

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