浅谈兽医实验室生物安全管理

随着生物技术的快速发展,在病原微生物研究、遗传基因工程、生物技术开发、生物制品研发、动物疫病防控方面,越来越多的兽医生物安全实验室,特别是高等级兽医生物安全实验室相继建立和投入使用.兽医实验室生物安全不仅关系公共卫生安全,还直接影响国民经济的正常发展,甚至关乎一个国家的社会稳定和安全.病原体逃逸出设施造成人和动物感染的事例时有报道,为确保医药、农业等行业中生物技术的健康迅速发展和应用,生物安全工作的研究和管理尤为重要,不可掉以轻心.     

高等级兽医生物安全实验室生物危害及其风险评估要素

<正>兽医生物安全实验室的主要工作内容是动物源性病原微生物样品的保存、处理、分离培养和鉴定等工作,因此实验室是病原体、未知病原因子、带病动物、易感人群和传播途径相对集中的特殊场所。部分动物尤其人兽共患病潜伏期动物潜在危害大,涉及个人健康、环境污染等。因此应在开展实验活动之前,对实验活动所操作的病原微生物以及实验活动本身进行评估,确保生物安全。兽医生物安全实验室,特别是高等级生物安全实验室的生物安全风险评估及风险控制,对确保实验室生物安全,保障     

全球生物安全与生物安全实验室的起源和发展

近年来全球陆续暴发的人类和动物疫病使各国愈发关注生物安全问题,生物安全实验室作为重要的生物安全科技支撑平台,其建设和管理也受到了各国的普遍重视。文章系统梳理了全球生物安全和生物安全实验室的起源与发展历程,重点分析了全球高等级生物安全实验室建设、管理等发展现状,并展望了全球生物安全实验室的发展前景,为我国生物安全实验室的建设和管理提供参考。     

兽医高等级生物安全实验室安全隐患分析及对策建议

正高等级生物安全实验室是指生物安全防护级别为三级(下文简称三级实验室)和四级(下文简称四级实验室)的生物安全实验室[1]。随着生命科学的日新月异,兽医高等级生物安全实验室在高危险度烈性病原体研究、新发突发传染病疫情防控、生物技术开发、转基因技术和遗传基因工程应用等方面所发挥的作用日益增强,各国政府对高等级实验室的建设和使用越来越广泛[2]。高等级生物安全防护实验设施是开展传染病研究、流行病学调查与研究、传染病     

高级别动物生物安全实验室气密性的探讨

高级别动物生物安全实验室气密性是保证实验室空气洁净度、室内实验人员和室外环境安全的重要手段。我国现行规范对高级别动物生物实验室各区域气密性有严格的规定。从实验室装修、围护结构设备安装以及气密性检测方面对实验室气密性进行了讨论,以期为我国高级别动物生物安全实验室的建设提供参考。     

欧洲高等级兽医生物安全实验室建设与生物安全管理

目前,我国高等级兽医生物安全实验室建设工作刚刚起步,同发达国家相比有很大差距,如何设计和建设,以及建设完后如何管理,如何安全地运行和使用等问题都摆到了我们面前。     

科技前沿

正中国首个自主设计建设高级别生物安全实验室投用8月7日,中国农业科学院哈尔滨兽医研究所举行了国家动物疫病防控高级别生物安全实验室认可证书授予仪式,标志着我国唯一的大动物生物安全四级实验室符合国家标准要求,开始投入运行。这是我国第一个完全自主设计、建设和管理运行的生物安全四级实验室。其建成运行,使我国具备开展所有已知重要传染病病原动物试验的能力,是我国生物安全保障能力建设的重要成就,     

论兽医生物安全实验室管理

<正>兽医生物安全实验室是指对病原微生物进行试验操作时所产生的生物危害具有物理防护能力的兽医实验室。多年来,随着国家动物疫病防控规划的实施,省、市、县各级动物疫控机构、兽医相关企业的生物安全实验室迅速发展起来,如何安全运行和有效管理兽医生物安全实验室,是实验室管理者、使用者以及有关主管部门需要重视的问题。1我国兽医生物安全实验室布局现状我国生物安全实验室建设起步相对较晚,法规和标准制定也较晚。SARS和高致病性禽流感疫情     

亨德拉病毒实验活动中安全风险评估

高致病性病原微生物实验活动中存在着诸多潜在的危险因素,笔者从生物安全角度出发,选定了亨德拉病毒为研究模型,对高致病性病原微生物实验活动中的安全风险进行评估,按照高等级生物安全实验室风险管理的要求,对感染性因子的特性、实验过程中的生物安全防护、意外安全风险等要素进行了风险评估,并提出了应急措施。为积极推进高等级实验室生物安全风险评估工作的开展,撰写高致病性病原微生物风险评估报告积累了素材。     

高级别兽医生物安全实验室的初步定量风险评估研究

[目的]探讨高级别兽医生物安全实验室的定量风险评估方法。[方法]对2005年以来通过认可的高级别兽医生物安全实验室,就设施、设备、管理体系和运行管理四大类风险要素开具的不符合项进行梳理、分析,通过模型构建,利用@Risk风险分析管理软件,对实验室运行中可能出现的事故概率进行初步量化评估。[结果]比较分析了有代表性的高级别兽医生物安全实验室现场评审发现的不符合项,按照比例权重依次为管理体系(30.3%)、设施(25.7%)、运行管理(18.2%)、其他(15.2%)和设备(10.6%)。构建了设施-事故、设备-事故、管理体系-事故和运行管理-事故的发生概率模型,通过50%百分位数统计量和平均值统计量分析各要素对事故概率的影响,由高到低依次为运行时间、运行管理、设施(平均值统计量分析时,在体系之后)、体系和设备;通过模型运算估计,就目前部分兽医实验室运行状况看,实验室年度内发生生物安全事故的概率最小为0.38%,平均发生事故的概率为2.5%,90%的可能发生事故的概率事件集中在0.8%~6.7%。严重疏于运行管理情况下,发生事故概率将有2.3%~2.5%升高至13.1%。[结论 ]高等级兽医生物安全实验室有多重要素影响其安全,应在注重设施建设、设备配置、体系编制基础上,加强日常运行的管理。     

IGF-1基因变异与犬体型及衰老的关系

体型较大往往是预测哺乳动物寿命较长的最佳指标之一,与此形成鲜明对比的是,有相当多的证据表明,在物种内部,体型较大的个体实际上寿命较短。这种体型增大的明显代价在犬上表现得尤为突出,人工选择导致犬种间体型相差近2个数量级,平均寿命相差2倍。胰岛素样生长因子1(IGF-1)是GH/IGF-1和IIS(insulin/IGF-1)通路中关键的调控基因,对各种生物的生长、发育、寿命以及代谢均具有重要作用。IGF-1与犬的体型和寿命都有着千丝万缕的联系,犬可作为一种研究IGF-1基因变异与体型大小和寿命之间关系的模型。论文综述了IGF-1基因变异与犬体型及衰老的研究,为抗衰老相关研究提供参考。     

昆明犬IGF-1基因CDS区克隆、生物信息学分析及组织表达研究

试验旨在系统研究昆明犬胰岛素样生长因子1（insulin-like growth factor-1，IGF-1）基因的结构和功能，揭示该基因的组织表达规律，为探讨工作犬体型及生长发育性状提供基本数据。以昆明犬为试验材料，运用分子克隆技术扩增IGF-1基因CDS区，应用生物信息学方法进行序列同源性比对并构建系统进化树；获得对应的氨基酸序列并分析蛋白理化特性及蛋白亚细胞结构、亲疏水性和磷酸化位点，利用SOPMA和SWISS-MODEL软件预测蛋白二级结构并构建蛋白质三级结构模型；同时利用实时荧光定量方法检测该基因在昆明犬各组织的表达情况。结果显示，昆明犬IGF-1基因CDS区长462 bp，编码153个氨基酸，其核苷酸序列与家犬的同源性较高（100%），与小鼠和鸡的同源性较低（82.6%和81.2%）。IGF-1蛋白分子质量和理论等电点分别为16.97 ku和9.36，属亲水性分泌型蛋白，无跨膜结构，存在信号肽区域（第1-48位氨基酸）；亚细胞定位分析表明，IGF-1蛋白主要分布于细胞核（56.5%）及线粒体（17.4%）中；磷酸化位点分析发现，IGF-1蛋白存在15个磷酸化位点；该蛋白的二级结构以α-螺旋和无规则卷曲为主。组织表达显示，IGF-1基因在昆明犬不同组织中均有表达，尤其在脾脏、肝脏及肾脏中呈现高表达。本试验结果为今后深入研究IGF-1基因功能提供了理论参考，并为深入探讨IGF-1基因对昆明犬生长发育性状研究提供一定的基础资料。     

Serum insulin-like growth factor type 1 concentrations in healthy dogs and dogs with spontaneous primary hypothyroidism

Circulating insulin-like growth factor type 1 (IGF-1) concentrations in dogs have been correlated with standard breed bodyweight (SBBW or breed size). Thyroid and somatotropic functions, which have common effects and regulatory mechanisms, were investigated in hypothyroid dogs. IGF-1 was measured in 495 adult healthy dogs (N) and in 220 primary hypothyroid dogs (HOT) with clinical and biological signs of primary hypothyroidism. IGF-1 was determined as a function of SBBW (kg): ≤15 (group A); 1540 (group D). In HOT dogs, median fT4 and c-TSH values were 9pmol/L and 1.5ng/mL, respectively. A significant correlation between bodyweight (BW) and IGF-1 was observed in both HOT and N dogs. The median IGF-1 value (ng/mL) was significantly higher (P<0.01) in HOT dogs compared to N in groups B, C and D (230 vs. 182; 316 vs. 230; 606 vs. 306 respectively). In conclusion, IGF-1 concentration should be interpreted in the context of SBBW in dogs and increases in spontaneous primary hypothyroidism. However, it remains unclear if this association is directly due to hypothyroidism or is the result of the weight gain accompanying hypothyroidism.     

Circulating concentrations of insulin-like growth factor-1 in dogs with naturally occurring mitral regurgitation

Insulin-like growth factor-1 (IGF-1), which mediates most effects of growth hormone, has effects on cardiac mass and function, and plays an important role in the regulation of vascular tone. In humans, an inverse relationship between degree of heart failure (HF) and circulating IGF-1 concentrations has been found in several studies. In dogs with HF, few studies have focused on IGF-1. We examined circulating IGF-1 concentrations in dogs with mitral regurgitation (MR) caused by myxomatous mitral valve disease. Study 1 included 88 Cavalier King Charles Spaniels (CKCSs) with a broad range of asymptomatic MR (median serum IGF-1: 76.7 microg/L; 25-75 percentile, 59.8-104.9 microg/L). As expected, standard body weight and percentage under- or overweight correlated directly with IGF-1. MR (assessed in 4 different ways) did not correlate with IGF-1. In study 2, 28 dogs with severe MR and stable, treated congestive HF had similar serum IGF-1 concentrations (median, 100.8 g/L; 25-75 percentile, 74.9-156.5 microg/L) as 11 control dogs (79.6 microg/L; 25-75 percentile, 64.1-187.4 microg/L; P = .84). In study 3, the plasma IGF-1 concentration of 15 untreated CKCSs with severe MR was 16.4 +/- 24.2 microg/L lower (P = .02) at the examination when decompensated HF had developed (80.8 +/- 30.9 microg/L) than at a visit 1-12 months earlier (97.2 +/- 39.8 microg/L), possibly in part due to an altered state of nutrition. The studies document that circulating IGF-1 concentrations are not altered before development of congestive HF in dogs with naturally occurring MR, but decrease by approximately 20% with the development of untreated HE In treated HF, circulating IGF-1 concentrations apparently return to within the reference range.     

犬体型大小相关基因的研究进展

犬作为人类最早驯化的家养动物,在人类发展史和现代生活中扮演着非常重要的角色,如伴侣、看护、导盲、军警用犬等。人工选择过程使得家犬产生了极大的表型多样性,其中体型性状是犬最显著的表型性状之一,属高遗传力性状。本文综述了犬体型性状相关基因IGF1、IGF1R、GHR1、GHR2、SMAD2、STC2、HMGA2、IGSF1、ACSL4及IRS4的研究进展,为未来工作犬体型遗传改良育种提供重要的靶点和依据,为进一步阐明人类及其他家养动物在体型上的遗传变异提供参考。     

撒坝猪胰岛素样生长因子-Ⅰ基因多态性及其与部分生长性状的关联分析

胰岛素样生长因子-Ⅰ(insulin-like growth factor-Ⅰ,IGF-Ⅰ)基因的表达产物对猪肌肉的生长有着重要的调控作用。以IGF-Ⅰ基因作为撒坝猪生长性状的候选基因,运用PCR-RFLP技术对其HhaⅠ位点的多态性进行了检测,并对IGF-Ⅰ基因的多态性与12个阶段体重、日增重和体尺性状的关系进行了分析。结果表明,撒坝猪IGF-Ⅰ基因的多态性较为丰富,产生AA、AB和BB 3种基因型,其基因型频率分别是0.3473、0.5149、0.1378,等位基因A和B的频率分别是0.6048、0.3952,且该基因座位处于Hardy-Weinberg平衡状态,撒坝猪IGF-Ⅰ基因的遗传变异对其生长性状有着显著的影响(P<0.05或P<0.01),其中,不同阶段体重和日增重的增效基因型为AB基因型,180日龄体尺性状的增效基因型多为BB基因型(部分性状表现为AB基因型最高)。结合前人的研究,初步推断IGF-Ⅰ基因可能是影响猪生长性状的一个主效基因或是一个与影响猪生长性状的数量性状座位(quantiative trait locus, QTL)连锁的基因。     

Serum insulin-like growth factor-1 measurements in dogs: performance characteristics of an automated assay and study of some sources of variation

The aim of this study was to evaluate the performance characteristics of an automated immunoassay for canine insulin-like growth factor 1 (IGF-1) measurement and to investigate the possible effects of some sources of variation, such as diurnal variations, feeding/fasting cycles, and glucocorticoid administration, in dogs. The immunoassay evaluated had an adequate analytical performance with intra- and inter-assay coefficients of variation (CVs) lower than 10%, linear regression equations with correlation coefficients of 0.9993 and 0.9988 after serial dilutions, and a limit of quantification of 7.1 ng/mL that was even lower than that reported by the manufacturer. The assay was significantly affected by hemolysis and lipemia producing a significant decrease in IGF-1 concentrations, but not by bilirubinemia.Serum IGF-1 concentrations did not show significant diurnal changes in fed or fasted dogs and were not affected by glucocorticoid administration.     

An association between lifespan and variation in insulin-like growth factor I receptor in sheep

Longevity in livestock is a valuable trait. When productive animals live longer, fewer replacement animals need to be raised. However, selection for longevity is not commonly the focus of breeding programs as direct selection for long-lived breeding stock is virtually impossible until late in the reproductive life of the animal. Additionally the underlying genetic factors or genes associated with longevity are either not known, or not well understood. In humans, there is evidence that IGF 1 receptor (IGF1R) is involved in longevity. Polymorphism in the IGF1R gene has been associated with longevity in a number of species. Recently, 3 alleles of ovine IGF1R were identified, but no analysis of the effect of IGF1R variation on sheep longevity has been reported. In this study, associations between ovine IGF1R variation, longevity and fertility were investigated. Polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) was used to type IGF1R variation in 1,716 New Zealand sheep belonging to 6 breeds and 36 flocks. Ovine IGF1R C was associated with age when adjusting for flock (present 5.5 ± 0.2 yr, absent 5.0 ± 0.1 yr, P = 0.02). A general linear mixed effects model suggested an association (P = 0.06) between age and genotype, when correcting for flock. Pairwise comparison (least significant difference) of specific genotypes revealed the difference to be between AA (5.0 ± 0.1 yr) and AC (5.6 ± 0.2 yr, P = 0.02). A weak negative Pearson correlation between fertility and longevity traits was observed (r = -0.25, P < 0.01). The finding of an association between variation in IGF1R and lifespan in sheep may be useful in prolonging the lifespan of sheep.     

A long life among ruminants: giraffids and other special cases

In order to investigate differences in the relative maximum longevity and other life history parameter between ruminant species, we collated data on mean body mass, maximum longevity, gestation period and newborn mass in wild ruminant and camelid species. Among ruminants, giraffids (giraffe Giraffa camelopardalis and okapi Okapia johnstoni) have particularly high longevities, long gestation periods, and low intrauterine growth rates. A particularly high absolute and relative longevity is also achieved by the anoa (Bubalus depressicornis), a member of the bovinae (cattle-type ruminants) and an insular dwarf (inhabiting the Indonesian island of Sulawesi). The fact that some (but not all) other small ruminants also achieve surprisingly high longevities leads to the hypothesis that extreme relative longevities in this group are an indication for secondary body size reduction.     

Statistical analysis regarding the effects of height and weight on life span of the domestic dog

This study was undertaken to determine the association between life spans and breed size in the dog, based upon data derived from the pet population. Seventy-seven American Kennel Club breeds were analyzed with data collected for more than 700 dogs. Multiple linear regression analysis was carried out with longevity as the dependent variable and height or weight as the independent variable. A negative correlation was observed between height and longevity (r=-0.603, p<0.05), and between weight and longevity (r=-0.679, p<0.05). Weight was the significant predictor of life span (p<0.001), revealing that breeds smaller by weight generally live longer than heavier breeds. These data form the ground work for investigations of aging utilizing the dog as a model and provide owners with a quantitative method for predicting lifespan of dog breeds, thereby aiding in pet selection.