闽西地区猪伪狂犬病病毒变异株gC基因克隆与序列分析

为了解闽西地区2013年以来猪伪狂犬病病毒(PRV)流行株的现状、分子生物学特征和遗传演化规律,收集了闽西不同地区的14株PRV分离株,并对其gC基因进行遗传变异分析。结果表明,14株PRV分离株与2012年以来国内不同省份分离的PRV变异株的核苷酸序列同源性为98.8%~99.7%,而与Bartha疫苗株核苷酸序列同源性为93.6%~93.8%。氨基酸多序列比对发现,14株PRV分离株和近年来国内分离的PRV毒株gC基因氨基酸序列同源性较高,有很多相同的特征性氨基酸替换,其中最有特征性的是aa52~aa61,aa63~aa69位和aa186~aa194位氨基酸的替换和插入。遗传进化树分析表明,14株PRV分离株与2012年以来国内不同省份分离的PRV株位于一个相对独立的大分支中,且形成了一个独立的亚分支,而与Bartha疫苗株存在很大差异,亲缘关系较远。抗原表位预测分析表明,PRV闽西分离株与Bartha疫苗株相比发生了抗原漂移,该差异是否预示闽西地区流行株正在向远离疫苗株的方向变异尚有待进一步研究。     

闽西地区新流行猪伪狂犬病病毒gD基因分子特征

本研究收集闽西不同地区的17株猪伪狂犬病病毒(PRV)分离株,并对其gD基因进行遗传变异分析,探讨2013—2015年闽西地区PRV流行株的现状、分子生物学特征和遗传演化规律。结果显示,17株PRV分离株与7株PRV国内分离株核苷酸及氨基酸序列的同源性较高均为99.0%~100.0%;与4株国外PRV经典株的核苷酸及氨基酸序列同源性相对较低,分别为98.6%~99.3%,97.0%~99.0%。核苷酸多序列比对结果显示,17株PRV分离株最具有特征性的变化是在831~836bp插入了连续的6个碱基CCGGCC,进而导致gD氨基酸序列275~276位增加了2个氨基酸RP。抗原性分析结果显示,这2个氨基酸还位于gD蛋白的抗原表位区内。遗传进化树分析结果显示,17株PRV分离株与7株国内分离株、韩国Yangsan株及美国Becker株位于一个相对独立的遗传分支内,亲缘关系较近;与Bartha及Kaplan 2株匈牙利分离株位于不同的大遗传进化分支中,亲缘关系较远。     

闽西地区猪伪狂犬病病毒变异株gE基因新变异序列鉴定

为了解2013-2014年我国闽西地区猪伪狂犬病病毒(PRV)流行株的现状、分子生物学特征和遗传演化规律,收集闽西不同地区的15株PRV分离株,并对其g E基因进行遗传变异分析。结果表明,15株PRV分离株与2012年以来国内不同省份分离的14株PRV变异株核苷酸及氨基酸序列的同源性较高,分别为98.7%~99.9%和97.1%~99.8%;而与15株PRV经典株的核苷酸及氨基酸序列的同源性相对较低,分别为97.0%~99.6%和94.6%~99.3%。氨基酸多序列比对发现,g E氨基酸序列最具特征性的变化是第48位和第496位各有1个天冬氨酸(D)的插入,该插入特征为PRV变异毒株的重要标志。同时,抗原性分析发现这2个位点还位于g E蛋白的抗原表位区内。遗传进化树分析结果表明,15株PRV分离株与近3年国内不同省份分离的PRV株位于一个相对独立的分支中,而与经典毒株处于不同的分支中,亲缘关系相对较远。以上结果表明PRV变异毒株已成为我国闽西地区主要的流行毒株。     

伪狂犬病病毒FS-2015株gE和gB基因序列分析

为了解猪伪狂犬病病毒（porcine pesudorabies virus,PRV）gB和gE基因变异及遗传演化情况,本研究针对PRV FS-2015野毒株,应用"蚀斑法"对组织病料中病毒进行三轮纯化,应用全长扩增引物对FS-2015株gB和gE基因进行全基因扩增,并对PCR产物进行测序和序列分析。结果显示,PRV FS-2015株的gB、gE基因与国内外PRV参考毒株的核苷酸同源性分别为97.0%~100.0%和97.5%~99.7%,氨基酸同源性分别为96.4%~100.0%和95.3%~99.7%。氨基酸变异位点分析表明,FS-2015株的gB和gE基因均有位点突变和缺失。遗传进化分析表明,FS-2015株与国内近几年分离的PRV变异株GY、ZJ01、HB1201、HN1201、JS2012、BJ-YT和BP属于同一分支,同源性较高,亲缘关系较近;与PRV经典株Kaplan、Becker、NIA3、Kolchis、Bartha和Yangsan株属于不同分支,同源性较低,亲缘关系较远。从PRV FS-2015毒株与国内外经典毒株和当前国内流行的变异毒株的分析结果可知,PRV FS-2015毒株发生了一定的变异,属于当前国内流行变异毒株。本研究结果为广东省伪狂犬病分子流行病学调查、伪狂犬病的防控和疫苗株挑选工作提供参考数据。     

伪狂犬病病毒FS-2015株gE和gB基因序列分析

为了解猪伪狂犬病病毒(porcine pesudorabies virus,PRV)gB和gE基因变异及遗传演化情况,本研究针对PRV FS-2015野毒株,应用"蚀斑法"对组织病料中病毒进行三轮纯化,应用全长扩增引物对FS-2015株gB和gE基因进行全基因扩增,并对PCR产物进行测序和序列分析。结果显示,PRV FS-2015株的gB、gE基因与国内外PRV参考毒株的核苷酸同源性分别为97.0%～100.0%和97.5%～99.7%,氨基酸同源性分别为96.4%～100.0%和95.3%～99.7%。氨基酸变异位点分析表明,FS-2015株的gB和gE基因均有位点突变和缺失。遗传进化分析表明,FS-2015株与国内近几年分离的PRV变异株GY、ZJ01、HB1201、HN1201、JS2012、BJ-YT和BP属于同一分支,同源性较高,亲缘关系较近;与PRV经典株Kaplan、Becker、NIA3、Kolchis、Bartha和Yangsan株属于不同分支,同源性较低,亲缘关系较远。从PRV FS-2015毒株与国内外经典毒株和当前国内流行的变异毒株的分析结果可知,PRV FS-2015毒株发生了一定的变异,属于当前国内流行变异毒株。本研究结果为广东省伪狂犬病分子流行病学调查、伪狂犬病的防控和疫苗株挑选工作提供参考数据。     

猪伪狂犬病毒流行株JL1株主要基因的遗传变异分析

为了解我国吉林省某地区猪伪狂犬病毒流行株JL1株的遗传变异情况,对PRV JL1株TK、gI、gD、gE和gB基因进行克隆测序,并与GenBank上发表的国内外毒株进行序列分析和遗传进化分析。结果显示,JL1株TK、gI、gD、gE和gB基因与国内外其他参考毒株的核苷酸同源性为99.6%~100%,96.4%~99.9%,98.8%~99.9%,98.1%~99.9%,98.4%~99.9%;氨基酸同源性为99.4%~100%,94.3%~99.7%,97.3%~99.8%,95.7%~99.5%,97%~99.7%。氨基酸多序列比对发现,PRV JL1株gE基因第48位和496位各插入一个天冬氨酸(D),与流行变异株的基因特征一致;同时gD基因、gI基因和gB基因分别有不同数量的氨基酸的插入和缺失。遗传进化分析表明,JL1株与国内近年来分离的流行毒株亲缘关系较近,尤其与JS-2012流行毒株亲缘关系相对最近,而与国外分离的经典毒株亲缘关系较远。通过对PRV JL1流行毒株的重要基因进行遗传变异分析,进一步了解PRV变异情况,可为当前流行的猪伪狂犬病防控以及疫苗的研究提供参考依据。     

一株猪伪狂犬病病毒变异株的分离鉴定

为了解广东省猪伪狂犬病病毒(pseudorabies virus,PRV)野毒株的基因变异及遗传演化的情况,本试验对广东佛山疑似暴发伪狂犬病的猪场采集的病料(脑、肺脏、扁桃体、肝脏、脾脏)进行了PCR鉴定,初步鉴定为PRV毒株后,将阳性病料接种非洲绿猴肾细胞(Vero),进行毒株传代培养,对分离毒株进行PCR检测及小鼠感染试验,证实该病毒为PRV,并命名为PRV FS-2015株;并对该毒株进行细胞病变观察、病毒TCID50测定、毒株gC和TK基因扩增及序列分析。结果显示,PRV FS-2015株TCID50为10-7.5/0.1mL。PRV FS-2015株的gC和TK基因序列与国内外PRV参考毒株进行同源性比对分析发现,其核苷酸序列同源性分别为95.8%~100.0%和99.4%~100.0%,氨基酸同源性分别为92.3%~100.0%和98.7%~100.0%。遗传进化分析表明,PRV FS-2015株与国内近几年分离的PRV变异株GY、ZJ01、HB1201、HN1201、JS2012、BJ/YT和BP属于同一分支,同源性较高,亲缘关系更近;但与PRV经典株Kaplan、Becker、NIA3、Kolchis、Bartha、Yangsan、Min-A、SS和SL株的同源性较低,基因变异较大,表明PRV FS-2015毒株属于近几年流行的变异株。本研究结果可为广东省伪狂犬病的防控工作和疫苗株的选择提供科学依据。     

一株猪伪狂犬病病毒变异株的分离鉴定

为了解广东省猪伪狂犬病病毒（pseudorabies virus，PRV）野毒株的基因变异及遗传演化的情况，本试验对广东佛山疑似暴发伪狂犬病的猪场采集的病料（脑、肺脏、扁桃体、肝脏、脾脏）进行了PCR鉴定，初步鉴定为PRV毒株后，将阳性病料接种非洲绿猴肾细胞（Vero），进行毒株传代培养，对分离毒株进行PCR检测及小鼠感染试验，证实该病毒为PRV，并命名为PRV FS-2015株；并对该毒株进行细胞病变观察、病毒TCID₅₀测定、毒株gC和TK基因扩增及序列分析。结果显示，PRV FS-2015株TCID₅₀为10^-7.5/0.1 mL。PRV FS-2015株的gC和TK基因序列与国内外PRV参考毒株进行同源性比对分析发现，其核苷酸序列同源性分别为95.8%~100.0%和99.4%~100.0%，氨基酸同源性分别为92.3%~100.0%和98.7%~100.0%。遗传进化分析表明，PRV FS-2015株与国内近几年分离的PRV变异株GY、ZJ01、HB1201、HN1201、JS2012、BJ/YT和BP属于同一分支，同源性较高，亲缘关系更近；但与PRV经典株Kaplan、Becker、NIA3、Kolchis、Bartha、Yangsan、Min-A、SS和SL株的同源性较低，基因变异较大，表明PRV FS-2015毒株属于近几年流行的变异株。本研究结果可为广东省伪狂犬病的防控工作和疫苗株的选择提供科学依据。     

猪伪狂犬病病毒GD株的生物学特性和gB基因进化分析

为了解猪伪狂犬病病毒(Pseudorabies virus,PRV)变异毒株的生物学特性和gB基因遗传变异特性,本研究对本实验室分离鉴定的变异株PRV GD的一步生长曲线和病毒对小鼠的毒力进行了研究,结果显示PRVGD毒株对KM小鼠的LD50为102.32TCID50。此外,对PRVGD株的gB基因进行了全长扩增,并对gB基因进行了测序和序列分析。研究结果显示,PRVGD株的gB基因与国内外PRV参考株的核苷酸同源性为98.3%～100%,氨基酸同源性为96.8%～99.9%。与经典毒株相比,PRV GD株的gB基因有位点插入、突变和缺失。基于gB基因的系统进化树分析发现,PRV GD株与国内近几年分离鉴定的PRV流行变异毒株如ZJ01、TJ、HeN1和JS-2012株位于同一分支上,同源性较高,亲缘关系较近。与国内外经典毒株均处于不同进化树分支,亲缘关系较远。本研究为PRV流行变异毒株的分子流行病学研究以及针对变异毒株的控制和新型疫苗的研发提供一定的参考价值。     

猪伪狂犬病病毒山西分离株gE全基因的遗传变异多样性及流行特点分析

为了研究当前山西省猪伪狂犬病(PR)的流行特点及遗传变异情况,对山西分离的3株猪伪狂犬病病毒(PRV)分离株(SXQX株、SXTG株和SXYP株)的gE全基因进行了扩增、克隆和测序(已被GenBank收录)。并将gE全基因序列和推导出的氨基酸序列与不同国家和地区的主要流行株进行了遗传变异多样性分析。结果显示,3株PRV分离株的gE全基因序列之间核苷酸及推导的氨基酸序列同源性分别为99.7%～100.0%和99.5%～100.%,与34株不同国家和地区的PRV参考株之间同源性分别为97.1%～100.0%和94.6%～100.0%,与欧美分离株的同源性分别为97.1%～97.6%和94.6%～95.7%,与2012年以来我国不同省份分离株的同源性分别为98.8%～100.0%和98.1%～100.0%,与2012年以前分离的经典毒株同源性分别为98.4%～99.7%和97.2%～99.3%。PRV gE全基因组序列的进化分析表明,3株PRV分离株属于亚洲基因群中2012年以来国内不同地区相继分离的PRV变异株群,而与2012年前分离的经典强毒株群亲缘关系较远,与欧美基因群亲缘关系最远。进一步进行核苷酸及氨基酸多序列比对分析发现,3株PRV分离株的gE基因均有一些位点发生了突变。本试验为近年来山西地区PRV的流行特点和PRV变异情况提供了依据。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.     

Mechanism of Action of Probiotic Function of Lactobacilli

乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.     

Evaluation of serum concentrations of biomarkers of skeletal metabolism and results of radiography as indicators of severity of osteochondrosis in foals

OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.