基于TMT技术的牦牛妊娠期血清蛋白质组学分析

为了探究妊娠期内牦牛血清的蛋白质组特征,筛选并分析差异表达蛋白质(DEPs)在牦牛妊娠维持中的作用。本研究选择9头4岁半至6岁龄健康状况良好且当年未产犊的自然发情母牦牛进行人工授精(AI)。在AI后第30、60及90天采集其血清样品,并对授配母牦牛进行妊娠诊断。依据诊断结果将采集的样品进行回顾性分组,即妊娠第1、2、3月组及未妊娠组(未妊娠组样品来自AI后90天经妊娠诊断确诊为未孕的牦牛个体),每组样品3个生物学重复。利用串联质谱标签(TMT)技术对牦牛血清样品的蛋白表达情况进行定量分析,筛选差异表达蛋白,并对其进行GO功能富集和KEGG通路等生物信息学分析,进一步筛选与牦牛妊娠维持相关的潜在蛋白。结果显示,在4组牦牛血清蛋白样品中共获得497个蛋白,以差异倍数≥1.2或≤0.83,且P<0.05为条件筛选,共筛选到68个差异蛋白。GO功能富集和KEGG通路分析显示,这些DEPs在代谢、免疫系统、粘附、生物调节等生物学过程以及补体与凝血级联、金黄色葡萄球菌感染等与炎症和免疫途径相关的通路有明显富集。值得注意的是,妊娠期各组间差异蛋白富集通路中均包含有PI3K-Akt、Rap1、M...     

云南省部分地区马属动物感染弓形虫血清学调查

为了解云南省马属动物弓形虫病流行情况,用夹心ELISA方法对云南省8个地区3种马属动物共969份血清样品进行弓形虫血清流行病学调查。结果弓形虫的抗体总阳性率为8.15%(79/969),其中马弓形虫抗体阳性率为3.03%(8/264),驴弓形虫抗体阳性率为8.92%(37/415),骡弓形虫抗体阳性率为11.72%(34/290)。不同地区马属动物血清样本弓形虫阳性率差异不显著(P>0.05),不同种类马属动物血清样品弓形虫阳性率差异极显著(P<0.01)。     

马属动物进化特征概述

马属动物丰富的化石材料,记录了其5 500万年的起源、迁徙、灭绝和进化,使其成为进化生物学的教研材料。马属动物于始新世时期出现于北美,称为始马。由于气候的变化,导致白令海峡水面的上升和下降,马属祖先曾陆续经由白令桥抵达欧亚古路,但是均没有留下现存物种后代。马属动物在北美持续演化,到上新世时,现代马属的最近祖先出现,即上新马。在接下来约3百万年间,马属动物经历快速的辐射进化,在不同的环境中占领生态位,形成包括马、驴和斑马3个主要亚属。促成现存马属物种在极短的时间内分离的主要原因,是马属典型的快速核型进化。随着马基因组精细图和驴基因组草图的完成,及分子生物学的研究,马属核型多样性的识别越来越精准。马属动物剧烈的染色体重排,让科学家们越来越相信,其异种杂交能够产生后代,后代没有繁殖能力的原因是马属动物基因组之间的高度相似,使得其杂种能够存活;但是由于核型的巨大差异,其杂种后代生殖细胞在减数分裂时,不能均衡的分配两套染色体,但这个猜测仍需要大量的试验来证实。     

马属动物顽固性肠便秘的手术治疗讨论

马属动物顽固性肠便秘又称肠阻塞、肠秘结、肠内容物停滞,是马属动物的常见内科病,也是多发的胃肠性腹痛病,多发生于马和骡,而驴发病较少.笔者采用手术治疗顽固性肠便秘62例,其中马8例、骡41例、驴13例,全部治愈,取得了满意的效果.通过投服泻剂、直肠破结等治疗方法均无效,遂采用手术治疗,现报告如下.     

驴dCG/LH受体CDS的克隆测序

研究比较2种马属动物的LH受体基因CDS,以驴卵巢黄体为实验材料,利用RT-PCR克隆测序得到驴dCG/LH受体基因的部分cDNA序列与马的相应序列进行对比。结果表明:马和驴在C-498T、A-1409G、G-1468T、C-1478T 4个位点上存在差异,其中C-498T、G-1468T导致(CTT)Leu-Phe(TTT)、(AGG)Arg-Met(ATG)的氨基酸变化,这两处氨基酸差异分别位于受体跨膜蛋白的胞外域和跨膜第2胞外域。     

驴品种之一——新疆驴

正驴是人们比较熟悉的畜种之一。具有肉、皮、药、乳、役等多种经济用途。目前全世界约有毛驴4 000万头,其中47%分布在亚洲。驴属于哺乳纲、奇蹄目、马科、马属,马和驴同属马属,但不同种,它们有共同的起源,驴的体型比马和斑马都小,第三趾发达,有蹄,其余各趾都已退化。驴的形象似马,多为灰褐色,不威武雄壮,它的头大,且耳朵长,胸部稍窄,四肢瘦弱,躯干虽短,但较长于四肢,因而体高和身长不相等,呈小     

抗菌肽作饲用抗生素代替品的应用

动物以抗菌肽、吞噬作用和蛋白质级联分解反应(如补体系统的级联反应)来消灭入侵的病原微生物，其中抗菌肽除了可直接发挥杀灭微生物的作用外．还具有引发吞噬和特异性蛋白质级联反应的趋化作用。近年来的研究表明，抗菌肽还具抗炎症功能(见图1)。     

朝阳市马属动物体寄生虫调查报告及防治

<正>朝阳市马属动物饲养量逐年增多,特别是由于驴的价格几年来居高不下,致使驴的饲养量增加极其显著。寄生虫病是危害马属动物的主要疾病,可造成消瘦、贫血、生长速度缓慢、腹泻、流产甚至死亡,为了掌握马属动物体寄生虫病防治的科学依据,笔者于2012~2015年用4年时间根据全市不同地区和自然状况及畜群分布特点,对马属动物进行了全面系统的寄生虫调查,调查选取有代表性的乡(镇)、村的一些养马(驴、骡)户、肉驴(马)     

根据线粒体3个基因确定马属动物系统发育关系

[目的]马属动物因其化石证据发掘充足,已经成为物种进化的最佳例证,但化石证据尚存在断层或者彼此难以区分等问题。旨在从分子水平上对马属动物的遗传分化、亲缘关系等方面的鉴定提供根本认识,为马属动物的保护和开发利用提供一些帮助,并为生物学教学提供基本的教学资料。[方法]采用分子生物学软件,通过比对在GenBank中提交的马属动物线粒体DNA(mtDNA)序列,检测单核苷酸突变(SNP)位点,并对检测到的单核苷酸突变进行系统发育分析。[结果]3个线粒体基因(COX1、Cytb和ND4)存在丰富的遗传变异,能够揭示出现存马属动物间的亲缘关系。[结论]3种基因构建的系统发育树的拓扑结构基本一致,都是白犀牛作为外群,马和普氏野马序列先聚在一起,3种亚洲野驴聚在一起,所有的斑马聚在一起,驴和斑马再聚在一起。     

一种驴和马及狐狸源性成分快速检测方法的研究

实验旨在建立快速检测驴、马和狐狸源性成分的方法。以驴、马和狐狸的线粒体保守序列16S r RNA基因为靶基因,设计通用引物和以不同荧光素标记的特异Taq Man探针,构建与通用引物共用的阳性扩增内标(Internal Amplification Control,IAC)作为PCR监控反应体系。通过优化PCR反应体系和条件,建立能同时检测驴、马和狐狸源性成分的四重实时荧光PCR方法。结果表明:利用驴、马、狐狸、牛、猪、羊、水貂、狗、鸡、鸭、大豆、小麦、玉米等物种检测没有交叉反应,证明该方法特异性高,准确性好。该方法模板DNA检测灵敏度为0.01 ng。本研究建立的驴、马和狐狸源性四重荧光实时PCR检测体系灵敏度高、特异性强,能有效鉴别动物产品中驴、马和狐狸的源性成分,也可用作相关动物皮张真伪的鉴别方法。     

马传贫驴白细胞弱毒疫苗株基质蛋白基因的克隆与表达

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码基质蛋白（p15)的基因，并在大肠杆菌中进行了表达，所表达的蛋白是一种可溶性的融合蛋白，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化，在间接ELISA和免疫印迹试验中，重组的基质蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应，这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制及在接种马体人免疫应答的研究中。     

Anaesthesia and analgesia of the donkey and the mule

The number of donkeys and mules throughout the world is stable, and awareness of their use and concern for welfare, pain recognition and treatment are receiving increasing veterinary interest. Therefore, accurate information about anaesthesia and analgesia in donkeys and mules is important to ever more equine practitioners. Since donkeys are physiologically and pharmacologically different from horses, knowledge on species specific aspects of anaesthesia and analgesia are very important. Mules combine elements from both donkey and horse backgrounds, leading to great diversity in size, temperament and body type. Physiologically, they seem to resemble horses more than donkeys. This review highlights the current knowledge on various anaesthetic and analgesic approaches in donkeys and mules. There is still much information that is not available about donkeys; in many circumstances, the clinician must use available equine information to treat the patient, while monitoring carefully to observe for differences in response to therapy compared to the horse.     

马传贫驴白细胞弱毒疫苗株跨膜蛋白主要免疫决定区基因的克隆与表达

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码跨膜蛋白主要免疫决定区(TMIR)的基因，并在大肠杆菌中进行了表达。所表达的融合蛋白有一部分是可溶的，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。在间接酶联免疫吸附试验(ELISA)和免疫印迹试验中，重组的TMIR蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应。这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制、接种马体内免疫应答及马传贫诊断的研究。     

Coagulation profiles of healthy Andalusian donkeys are different than those of healthy horses

Background: Coagulation disorders are frequently diagnosed, especially in hospitalized equidae, and result in increased morbidity and mortality. However, hemostatic reference intervals have not been established for donkeys yet. Objectives: To determine whether the most common coagulation parameters used in equine practice are different between healthy donkeys and horses. Animals: Thirty‐eight healthy donkeys and 29 healthy horses. Methods: Blood samples were collected to assess both coagulation and fibrinolytic systems by determination of platelet count, fibrinogen concentration, clotting times (prothrombin time [PT] and activated partial thromboplastin time [aPTT]), fibrin degradation products (FDP) and D‐Dimer concentrations. Results: PT and aPTT in donkeys were significantly (P < .05) shorter than those of horses. In contrast, FDP and D‐Dimer concentrations were significantly (P < .05) higher in donkeys than in horses. Conclusions and Clinical Importance: The coagulation parameters most commonly determined in equine practice are different in donkeys compared with horses. Thus, the use of normal reference ranges reported previously for healthy horses in donkeys might lead to a misdiagnosis of coagulopathy in healthy donkeys, and unnecessary treatments in sick donkeys. This is the first report of normal coagulation profile results in donkeys, and further studies are warranted to elucidate the physiological mechanisms of the differences observed between donkeys and horses.     

Differential responses of Equus caballus and Equus asinus to infection with two pathogenic strains of equine infectious anemia virus

Most in vivo studies with equine infectious anemia virus (EIAV) have been performed in horses and ponies (Equus caballus) with little published information available detailing the clinical responses of donkeys (Equus asinus) to infection with this virus. Consequently, donkeys were inoculated with two strains of EIAV (EIAV(PV) and EIAV(WY)) which have been documented to produce disease in E. caballus. Four ponies, 561, 562, 564 and 567 and two donkeys, 3 and 5 were infected with EIAV(PV) and one horse (94-10) and one donkey (4) were infected with EIAV(WY). Although the horse and ponies all experienced clinical signs of disease, which in some cases were severe, the donkeys remained asymptomatic throughout a 365-day observation period, except for mild transient reductions in platelet counts. The results from serological assays, virus isolation from plasma and detection of plasma-associated viral RNA by RT-PCR, indicated that initial replication of EIAV(PV) and EIAV(WY) was lower in donkeys than in horses and ponies. This conclusion was confirmed using competitive RT-PCR, in which viral RNA levels in the plasma of EIAV(PV)-infected ponies was up to 100,000-fold higher than in infected donkeys during the first 20 days post-infection (dpi). Similar results were obtained in the EIAV(WY)-infected animals, in which viral RNA burdens in the donkey at 20 dpi were 1000-fold less than in the horse. However, infection of donkey and horse monocyte-derived macrophage cultures with EIAV(PV) demonstrated that these cells in vitro were equally susceptible to virus-induced cytopathic effects and yielded similar levels of progeny virus. This result suggests that factors other than host cell permissiveness mediate the clinical differences observed between horses and donkeys infected with EIAV(PV) or EIAV(WY).     

马、驴主要经济性状功能基因研究进展

马、驴是重要的草食家畜,在人类的历史变迁和生产生活中扮演重要角色。马、驴的初始功能主要以役用为主,多用于乘骑或驮运物品。随着机械化程度和交通方式的进步,其役用功能逐渐降低甚至消失。现代马产业主要以竞技、休闲娱乐及副产品加工为主,而驴产业则以皮、肉、乳及其生物制品的开发利用为主。马经济性状主要包括体型、毛色、竞赛能力、疾病、极端环境适应能力等,在驴上更关注生长、皮用和泌乳等性状。随着基因组学和生物信息学的发展,马、驴主要经济性状相关基因的发掘更为有效和精准：如与马体重、体尺相关的基因被定位于LCORL/NCAPG基因区域;MSTN基因与骨骼肌的发育相关,进而影响马竞赛性能;与设特兰矮马、德保矮马矮小性状相关的主要基因分别为HMGA2和TBX3基因;ACAN基因突变会导致设特兰矮马侏儒;DMRT3基因突变影响马的步态特征;KIT基因与白斑毛表型相关,MC1R基因是控制栗色毛的主要基因,ASIP基因与黑色毛相关;EDNRB基因突变会导致致死白色马驹综合征;EPAS1基因和线粒体NADH6基因在高原适应性进化中起重要作用。作者对马、驴主要经济性状相关功能基因的研究进展进行综述,并对功能基因组学研究进行展望,以期为今后开展马、驴分子遗传育种研究提供借鉴与参考。     

An investigation of equine infectious anaemia infection in the central Anatolia region of Turkey

In this study, 162 horses, 80 donkeys and 51 mule serum samples were collected in Konya city. Additionally, 64 horse serum samples from Ankara and 49 samples from Kayseri city were included in the study. A total of 406 serum samples were examined by agar gel immunodiffusion (AGID) and enzyme-linked immunosorbent assay (ELISA) for antibody to equine infectious anaemia virus (EIAV) and no positive result was detected.     

德州驴与泌阳驴睾丸组织mRNA与microRNA表达鉴定与分析

试验旨在探究驴睾丸组织中mRNA和microRNA的表达模式，为中国地方驴的转录组研究提供遗传数据。利用RNA-Seq和生物信息学方法分析了中国地方驴睾丸组织表达基因和microRNA，并分析了泌阳驴和德州驴品种间差异表达基因及microRNA。结果表明，mRNA平均clean reads为57837506条，Small RNA平均clean reads为9952015条，分别占原始数据的96.34%和88.86%。在驴睾丸组织中共发现24751个表达基因及1074个表达microRNAs，其中，睾丸组织表达量较高的基因为LOC106836782、XLOC_066401、HSP90AA1、TNP1和COF2，表达量最高的microRNAs为eca-miR-143、eca-miR-21、eca-miR-99a、eca-miR-148a和eca-miR-26a。对泌阳驴与德州驴睾丸组织进行差异表达分析共发现102个差异表达基因及2个差异表达microRNAs，但未富集到功能条目，说明泌阳驴和德州驴品种间差异不明显。本研究提供了驴睾丸组织mRNA及microRNA表达模式和转录组学数据，并鉴定了泌阳驴与德州驴睾丸组织的差异表达mRNA和microRNA，对驴的遗传资源研究具有一定的科学意义。     

Pharmacology and therapeutics in donkeys

Therapeutics are often administered to donkeys based on dosage and intervals recommended for horses because very few drugs have donkey‐specific label indications. Yet differences between donkeys and horses in drug distribution, metabolism and elimination have been noted for most therapeutic agents studied. These differences can be partially explained by the donkey's unique physiology. Since their ancestors evolved in a desert environment, the modern donkey exhibits qualities that allow them to tolerate dehydration better than the horse and recover more quickly from its effects. Fluid balance and body water compartment partitioning differ from the horse and may have implications regarding drug distribution. Since donkeys are preferential browsers, differences in diet may have influenced evolutionary differences in metabolic disposition of drugs. It is important to acknowledge these differences when designing dose regimes for donkeys based on horse protocols in order to avoid either lack of efficacy or toxicity.     

Comparative pharmacokinetics of meloxicam in clinically normal horses and donkeys

OBJECTIVE: To determine the disposition of a bolus of meloxicam (administered IV) in horses and donkeys (Equus asinus) and compare the relative pharmacokinetic variables between the species. ANIMALS: 5 clinically normal horses and 5 clinically normal donkeys. PROCEDURES: Blood samples were collected before and after IV administration of a bolus of meloxicam (0.6 mg/kg). Serum meloxicam concentrations were determined in triplicate via high-performance liquid chromatography. The serum concentration-time curve for each horse and donkey was analyzed separately to estimate standard noncompartmental pharmacokinetic variables. RESULTS: In horses and donkeys, mean +/- SD area under the curve was 18.8 +/- 7.31 microg/mL/h and 4.6 +/- 2.55 microg/mL/h, respectively; mean residence time (MRT) was 9.6 +/- 9.24 hours and 0.6 +/- 0.36 hours, respectively. Total body clearance (CL(T)) was 34.7 +/- 9.21 mL/kg/h in horses and 187.9 +/- 147.26 mL/kg/h in donkeys. Volume of distribution at steady state (VD(SS)) was 270 +/- 160.5 mL/kg in horses and 93.2 +/- 33.74 mL/kg in donkeys. All values, except VD(SS), were significantly different between donkeys and horses. CONCLUSIONS AND CLINICAL RELEVANCE: The small VD(SS) of meloxicam in horses and donkeys (attributed to high protein binding) was similar to values determined for other nonsteroidal anti-inflammatory drugs. Compared with other species, horses had a much shorter MRT and greater CL(T) for meloxicam, indicating a rapid elimination of the drug from plasma; the even shorter MRT and greater CL(T) of meloxicam in donkeys, compared with horses, may make the use of the drug in this species impractical.