边界病研究进展

边界病(border disease,BD)是由边界病毒(BDV)引起绵羊和山羊的一种病毒性疾病,临床症状表现为母羊不孕、流产和死胎,以及羔羊体型异常,多毛,震颤,因此又称"多毛震颤病","茸毛症"。该病的主要传播途径是垂直传播,持续性感染的羔羊是该疾病在绵羊中传播的潜在主要传染源。山羊相对绵羊感染症状相对较轻,主要表现为流产。目前尚无公认有效的BD疫苗,做好生物安全防控,严格饲养管理,及时对疑似染病动物特别是持续性病毒血症绵羊的检测和淘汰是防治本病的重要措施。现对该病的流行现状、病原学、流行病学、鉴别诊断和预防控制等方面的研究进展进行综述,旨在为研究人员及养殖户加强对BD的认识,做好疾病的相关预防工作,减少经济损失提供参考。     

我国羊隐孢子虫感染情况及危险因素

隐孢子虫是可以感染小反刍动物的肠道原生生物,具有潜在的公共卫生学问题。隐孢子虫可引起人和动物的腹泻和肠道疾病,严重情况下会出现死亡。被感染的动物可能是人畜共患型隐孢子虫的宿主,可引起公共卫生风险、农场利润减少和动物福利等问题。隐孢子虫病已被报道是新生牛、羊等反刍动物腹泻和死亡的一个重要原因,认为是新生羔羊腹泻的第二大诱因,仅次于轮状病毒。羊隐孢子虫病在世界范围内都有发现,不同程度地威胁着人和动物的身体健康,基于此,现就隐孢子虫对羊感染的危险因素和常用检测方法,以及我国羊感染的现状进行综述,为其防控提供参考。     

妊娠后期营养限制对蒙古绵羊胎儿生物原胺类神经递质的影响

本试验旨在研究妊娠后期营养限制对蒙古绵羊胎儿生物原胺类神经递质的影响。选择健康、体况相近的蒙古绵羊18只,对其进行同期发情、配种后,从妊娠第90天开始,随机分为3个组:营养限制1组[NG1组,n=6,代谢能(ME)=0.175 MJ/(kg BW 0.75·d)]、营养限制2组[NG2组,n=6,ME=0.330 MJ/(kg BW 0.75·d)]和对照组[CG组,n=6,ME=0.670 MJ/(kg BW 0.75·d)],在妊娠第140天时屠宰,取各组胎儿及其脑组织和血液。结果显示:NG1组胎儿的脑重较CG组显著增高(P<0.05),并且NG1组胎儿脑中5-羟色胺(5-HT)(P<0.01)、肾上腺素(EPI)(P<0.05)含量显著或极显著高于CG组,去甲肾上腺素(NA)的含量极显著低于CG组(P<0.01)。NG2组胎儿脑中5-HT含量极显著高于CG组(P<0.01),而NA(P<0.01)、多巴胺(DA)(P<0.05)含量显著或极显著低于CG组。另外,NG1组胎儿血浆中5-HT(P<0.01)、EPI(P<0.01)、NA(P<0.05)、DA(P<0.01)含量显著或极显著低于CG组;NG2组胎儿血浆中5-HT(P<0.01)、NA(P<0.01)、DA(P<0.05)含量显著或极显著低于CG组。由此得出,妊娠后期营养限制使得蒙古绵羊胎儿脑中5-HT、EPI含量升高和NA含量降低,血浆中5-HT、EPI、NA、DA含量降低。     

饲粮添加氨基酸锌对育肥羊屠宰性能、肉品质及血液和组织中微量元素含量的影响

本试验旨在研究氨基酸锌对育肥羊屠宰性能、肉品质及血液和组织中微量元素含量的影响。选择4月龄、体重为(29.00±0.61)kg的健康滩湖杂F1代公羊48只,随机分为4组,每组12只。对照组饲粮锌含量为52.80 mg/kg(硫酸锌),其中不含硫酸锌的锌含量为18.02 mg/kg,试验Ⅰ组、试验Ⅱ组、试验Ⅲ组试验饲粮锌含量分别为对照组饲粮的1/2、1和2倍(氨基酸锌),预试期7 d,正试期60 d。结果表明:饲粮添加氨基酸锌显著提高了育肥羊平均日增重(P<0.05),以试验Ⅱ组平均日增重最高,达到283.59 g/d。饲粮添加氨基酸锌对育肥羊屠宰性能、肉质理化性状无显著影响(P>0.05),但随着饲粮氨基酸锌添加量增加,试验组肉骨比、背膘厚、眼肌面积先升高后降低,GR值、肌内脂肪含量升高,剪切力降低;试验组血液锌含量均高于对照组,在同等锌添加量条件下,试验Ⅱ组血清锌含量极显著高于对照组(P<0.01);各组间背最长肌、肾脏、心脏、肝脏及肺脏组织的锌含量差异不显著(P>0.05),但氨基酸锌添加量过高会影响肌肉、肾脏、心脏、肺脏中铁沉积及肝脏中铜沉积。综上所述,饲粮添加氨基酸锌可促进育肥羊生长,改善肉品质;在锌含量为18.02 mg/kg的基础饲粮中添加52.80 mg/kg锌(氨基酸锌)效果最佳。     

Toll-like receptor signaling is changed in ovine lymph node during early pregnancy

Toll-like receptors (TLRs) participate in regulation of adaptive immune responses, and lymph nodes play key roles in the initiation of immune responses. There is a tolerance to the allogenic fetus during pregnancy, but it is unclear that expression of TLR signaling is in ovine lymph node during early pregnancy. In this study, lymph nodes were sampled from day 16 of nonpregnant ewes and days 13, 16, and 25 of pregnant ewes, and the expressions of TLR family (TLR2, TLR3, TLR4, TLR5 and TLR9), adaptor proteins, including myeloid differentiation primary-response protein 88 (MyD88), tumor necrosis factor receptor associated factor 6 (TRAF6), and interleukin-1-receptor-associated kinase 1 (IRAK1), were analyzed through real-time quantitative polymerase chain reaction, Western blot, and immunohistochemistry analysis. The results showed that mRNA and protein levels of TLR2, TLR3, TLR4, TRAF6, and MyD88 were upregulated in the maternal lymph node, but TLR5, TLR9, and IRAK1 were downregulated during early pregnancy. In addition, MyD88 protein was located in the subcapsular sinus and lymph sinuses. Therefore, it is suggested that early pregnancy induces changes in TLR signaling in maternal lymph node, which may be involved in regulation of maternal immune responses in sheep.     

AANAT基因在滩羊不同繁殖时期卵巢中的转录差异及DNA甲基化程度分析

试验旨在检测5-羟色胺-N-乙酰基转移酶（AANAT）基因在绵羊休情季节和繁殖季节（卵泡期和黄体期）卵巢组织中的转录差异,并分析转录差异是否由DNA甲基化修饰程度改变所导致。试验采用自然环境条件和饲养管理一致,且体重差异在0.5 kg范围内的空怀母滩羊作为试验动物,采集其休情期、卵泡期和黄体期（每个时期3只）的卵巢组织,采用SYBR染料法进行实时荧光定量PCR检测AANAT基因在滩羊不同繁殖时期卵巢组织中的转录水平。随后针对转录水平有差异的两个时期（休情期和卵泡期）的样本,利用MethPrimer 2.0在线软件预测AANAT基因启动子区和第一外显子区的CpG岛;用重亚硫酸盐测序法（BSP法）检测AANAT基因启动子区及第一外显子区的甲基化程度。试验结果显示,滩羊休情期卵巢组织中AANAT基因转录水平显著低于卵泡期的AANAT基因转录水平（P<0.05）,休情期与黄体期滩羊卵巢组织中AANAT基因的转录水平差异不显著（P>0.05）。滩羊卵巢组织中AANAT基因启动子区上存在着一个长度为173 bp的CpG岛,第一外显子区存在着一个长度为118 bp的CG岛。然而,两个甲基化岛区内的单个CpG位点甲基化程度在滩羊休情期和卵泡期之间均不存在显著差异,暗示AANAT基因的表达受甲基化修饰外的因素调控。本研究结果可为进一步探讨AANAT基因在季节性发情和卵泡成熟中的功能提供参考资料。     

绵羊乳酪蛋白血管紧张素转换酶抑制肽的制备与鉴定

以绵羊乳酪蛋白为原料,采用中性蛋白酶水解制备生物活性肽,研究酶解时间对酶解产物水解度和血管紧张素转换酶（angiotensin converting enzyme,ACE）抑制率的影响,利用液相色谱-质谱联用技术对水解产物进行分析和鉴定,筛选具有潜在ACE抑制作用的生物活性肽。结果表明：酶解时间达到300 min时,水解度最高值达9.65%,ACE抑制率为84.55%;当ACE抑制率达到50%时,所需肽段YYQQRP浓度为5～10 μmol/L;利用超高效液相色谱飞行时间质谱仪对酶解后的多肽进行序列分析,     

The utility of a novel formulation of alfaxalone in a remote delivery system

ObjectiveTo quantify induction time, reliability, physiological effects, recovery quality and dart volume of a novel formulation of alfaxalone (40 mg mL^?1) used in combination with medetomidine and azaperone for the capture and handling of wild bighorn sheep.Study designProspective clinical study.AnimalsA total of 23 wild bighorn sheep (Ovis canadensis) in Sheep River Provincial Park, AB, Canada.MethodsFree-ranging bighorn sheep were immobilized using medetomidine, azaperone and alfaxalone delivered with a remote delivery system. Arterial blood was collected for measurement of blood gases, physiologic variables (temperature, heart and respiratory rates) were recorded and induction and recovery length and quality were scored.ResultsData from 20 animals were included. Administered dose rates were alfaxalone (0.99 ± 0.20 mg kg^?1; 40 mg mL^?1), azaperone (0.2 ± 0.04 mg kg^?1; 10 mg mL^?1) and medetomidine (0.16 ± 0.03 mg kg^?1; 30 mg mL^?1). The mean drug volume injected was 1.51 mL. The median (range) induction time was 7.7 (5.8–9.7) minutes, and recovery was qualitatively smooth.Conclusions and clinical relevanceAn increased concentration formulation of alfaxalone was administered in combination with medetomidine and azaperone, and resulted in appropriate anesthesia for the capture and handling of bighorn sheep. The dart volume was small, with potential for reducing capture-related morbidity.     

Cryopreservation of ram semen: Manual versus programmable freezing and different lengths of equilibration

The aim of our study was to examine effects of the length of semen equilibration as well as two freezing techniques on ram sperm post-thaw quality. The ejaculates of Wallachian sheep rams (n = 12) were collected by an electro-ejaculation, equilibrated in a Triladyl® (0, 2, 4, 6, and 8 h) containing glycerol and egg yolk and frozen by programmable freezing (PF) or manual freezing (MF). After thawing, sperm samples were subjected to the motility (computer-assisted sperm analysis [CASA]), viability (SYBR-14/PI), and fertilizing ability (FA) (in vitro penetration/fertilization test on bovine oocytes) assays. It was found that the equilibration of 6 h (E-6) ensured higher post-thaw sperm motility and progressive movement compared with other lengths tested, irrespective of a freezing technique. The E-6 sperm viability did not differ between PF and MF but was lower (P < 0.05) than control. Sperm FA (E-6) was similar in PF (60.44%) and MF (62%) but slightly lower than in fresh (72.8%). Our data demonstrate that the use of MF was comparable with PF, which can be applied in the field conditions without need in a piece of cost-expensive equipment, which can greatly benefit the gene bank of animal genetic resources.