51-Chromium Labeled Erythrocyte Half-time Disappearance from Miniature Swine

Half-time disappearance of ⁵¹Cr-laeled erythrocytes was studied in 39 Sinclair miniature swine at 20 to 46 days of age and again at 79 to 109 days of age. Mean erythrocyte half-time disappearance, disregarding age, was 21.29 (±0.77) days for males and 22.04 (±0.81) days for females. The half-time difference between males and females was not statistically significant (P<0.05). At 20 to 46 days of age the mean erythrocyte half-time disappearance was 19.56 (±1.41) days. The same pigs, tested at 79 to 109 days of age, had a mean erythrocyte half-time disappearance of 23.95 (±1.94) days. There was a significant difference (P<0.01) in erythrocyte half-time disappearance from miniature swine as the result of age and also for age within sex. The grand mean erythrocyte half-time disappearance, for all the miniature swine tested, was 21.67 (±0.54) days. The estimate of heritability was 0.61 for dam effect at 20 to 46 days of age and 0.06 at 79 to 109 days of age. Sire estimate of heritability was 0 at both age groups. It was concluded that the large heritability coefficient found only for dam component in young nursing pigs was the result of maternal environment rather than additive genetic variance.     

Serum Biochemical and Hematological Parameters of Sinclair(S-1) Miniature Sows During Gestation and Lactation

Venous blood samples were collected from 29 Sinclair(S-1) miniature sows at 14, ten, six and two weeks prior to parturition and two, four and six weeks postpartum to determine the effect of pregnancy and lactation upon 19 serum biochemical and 12 hematological parameters. During gestation, the levels of serum cholesterol, blood urea nitrogen and alpha₁-globulin, as well as packed cell volume and hemoglobin concentration, decreased; whereas, the level of serum beta-globulin increased. During lactation, the concentrations of serum glucose, total protein, albumin, beta-globulin, calcium, sodium and hemoglobin, as well as packed cell volume, decreased; whereas, the concentration of serum cholesterol and the activity of serum alkaline phosphatase increased.     

The Electrocardiogram of Conventional and Miniature Swine (Sus scrofa)

Standard, augmented limb leads and lead V₁₀ (representing the Z axis) taken in sequence and three semi-orthogonal leads (I, aVF, and V₁₀) taken simultaneously were recorded from 43 healthy pigs. Records were analyzed for rate, rhythm, interval duration and component amplitudes. The wave form of QRS complexes were analyzed in all leads studied. P, QRS, and T vectors were calculated for the mean dorsal (frontal), sagittal, and transverse planes. study of orthogonal leads in right sternal recumbency indicated that ventricular activation is spatially oriented dorsad, sinistrad and slightly caudad. No essential difference was noticed in the conventional and miniature swine. Ventricular fibrillation induced electrically in four older pigs was a progressive, terminal event; in two newborn piglets, ventricular fibrillation induced several times was terminated in each case by spontaneous recovery to sinus rhythm.     

鸽Ⅰ型副黏病毒(S-1株)灭活疫苗免疫剂量研究

为了确定鸽Ⅰ型副黏病毒(pigeon paramyxovirus-Ⅰ,PPMV-Ⅰ)(S-1株)灭活疫苗的免疫剂量。本试验采用3批PPMV-Ⅰ(S-1株)灭活疫苗实验室制品以不同剂量肌肉注射免疫30日龄低抗体幼龄鸽(HI抗体≤2)和120日龄低抗体青年鸽(HI抗体≤2),分别在免疫后第14,28天对试验鸽进行攻毒,对试验鸽进行临床症状、抗体检测和攻毒保护测定。结果显示,该疫苗对低抗体鸽有保护作用,对30日龄幼龄鸽和120日龄青年鸽的最小免疫剂量分别为0.05,0.2mL/只。因此,为了确保疫苗的免疫效果,在PPMV-Ⅰ(S-1株)灭活疫苗制造及检验规程中疫苗用法用量规定:幼龄鸽肌肉注射,0.2mL/只;青年鸽和种鸽肌肉注射,0.5mL/只。     

地高辛标记核酸探针检测重组人白细胞介素1受体颉颃剂中DNA残余量的研究

试验旨在建立重组人白细胞介素1受体颉颃剂(interleukin-1 receptor antagonist,IL-1ra)原液中宿主菌DNA残余量的检测方法。以重组白细胞介素1受体颉颃剂工程菌基因组DNA为模板,优化试验条件,借助超声波手段,制备地高辛标记的核酸探针, 并以此探针进行狭缝杂交及显色反应。结果显示,在24 批次重组人IL-1ra原液中,每人份使用剂量的宿主DNA残余量均小于10 ng。结果表明,该方法灵敏度较高、特异性较强、重复性较好、操作步骤安全且较为方便,可用于重组人IL-1ra原液及半成品的DNA残余检测。     

H1N1猪流感病毒环介导等温扩增快速检测方法的建立

目的:建立H1N1猪流感病毒环介导等温扩增(LAMP)快速检测方法。方法:从GenBank中获得H1N1猪流感病毒血凝素(HA)、神经氨酸酶(NA)基因序列,应用DNAStar软件MegAlign程序分析其序列,利用Primer ExplorerV4软件在序列保守区域设计LAMP引物,即外引物和内引物,同时以H1N1猪流感病毒的cDNA作为阳性模板,对试验中的几个反应条件进行优化。结果:LAMP检测方法对H1N1猪流感病毒的灵敏度达到4～6个拷贝,其引物对于H9亚型禽流感病毒、猪瘟病毒和猪圆环病毒均无非特异性扩增,表现出良好的特异性。结论:建立的H1N1猪流感病毒环介导等温扩增快速检测方法灵敏度高、特异性强、重复性好,为快速检测猪流感病毒提供了新方法和新思路。     

塞北兔红细胞酯酶(Es-1,Es-3)遗传多态性研究

采用垂直板聚丙烯酰胺线性浓度梯度凝胶电泳法,对216只塞北兔红细胞酯酶(Es-1,Es-3)的遗传多态性进行了检测。结果表明,在塞北兔群体中,红细胞酯酶(Es-1,Es-3)均呈多态,各有3种表型,受2个共显性等位基因A和B控制。X2检验结果表明,塞北兔群体在Es-1和Es-3位点处于Hardy-weinberg平衡状态(P>0.05)。     

猪源牛病毒性腹泻病毒的流行初探

根据牛病毒性腹泻病毒(BVDV)代表株NADL的NS5B基因序列,设计合成2对引物进行套式PCR,检测从浙江、安徽、湖南、江西、广西、辽宁等地采集的43份猪病料中BVDV的流行情况,结果有7份病料可以扩增出360bp的特异性条带,阳性率为16.3%。为追踪BVDV在猪体中的流行来源,我们又对细胞培养用国产及进口犊牛血清进行了检测,结果也能扩增到目的条带。对这些病料和血清中扩增出的目的条带进行克隆、测序,并用DNASTAR软件分析,结果表明这些目的片段均为BVDV序列,这些不同BVDV序列可以分为两个亚群,ZJ133、HN386、LN247、NCS-J属于BVDV-Ⅰa亚型,ZJ114、AH138、JX60、GX96、NCS-G、DZ属于BVDV-Ⅰb亚型。这些毒株与BVDVI型间的同源性为80.3%-98.6%,与BVDV-Ⅱ型的同源性为72.2%-74.4%。本文研究结果说明我国猪群中BVDV感染情况已经比较严重,应该予以重视。     

H1N1猪流感广东株血凝素基因的克隆与序列分析

用RT-PCR方法扩增H1N1亚型猪流感病毒广东分离株A／Swine／GuangDong／711／2001HA基因，对其进行了克隆和测序。结果显示，HA基因全长1771bp．共编码579个氨基酸。A／Swine／Guang Dong／711／2001HA基因编码的氨基酸序列中有8个糖基化位点，5个位于HAl基因的第27、28、40、104和287位点，3个位于HA2基因的21、153和213位点。与H1N1亚型猪流感经典毒株比较后发现，血凝素糖基化位点并不是高度保守的。将A／Swine／Guang Dong／711／2001与自Gen Bank读取的1918年人流感毒株A／South Carolina／1／18、1991年人流感毒株A／MD／12／91和1997年猪流感毒株A／Swine／Wisconsin／238／97等进行核苷酸同源性比较分析，A／Swine／Guang Dong／711／2001与A／SouthCarolina／1／18、A／MD／12／91和A／Swine／Wisconsin／238／97等毒株的核苷酸同源性分别为93．9％、94．7％和93．9％。从系统发生树来看，A／Swine／Guang Dong／711／2001与A／South Carolina／1／18和A／Swine／Wisconsin／238／97的亲缘关系相近。     

H1N2亚型猪流感病毒A／Swine／Guangdong／1／06分离鉴定和对猪致病性研究

2006年5月从广东某大型猪场采集具有流感症状保育猪鼻拭子共98份,无菌常规处理猪鼻拭子后接种MDCK细胞,分离到4株流感病毒.用血凝抑制和PCR方法鉴定均为H1N2亚型.挑选其中一株A/Swine/Guangdong/1/06(H1N2),进行全基因组测序,并与GenBank中相关序列进行比较.核苷酸同源性分析表明:A/Swine/Guangdong/1/06(HIN2)与A/Swine/Guangxi/13/06不同基因之间同源性为96.6%～98.1%.以106EID50的剂量,将H1N2病毒鼻腔感染35日龄仔猪,结果表明H1N2亚型流感病毒可以感染猪上呼吸道.但不表现临床症状.本研究结果对于揭示H1N2亚型猪流感流行规律和病毒的致病机理具有一定的意义.     

哈尔滨大白兔红细胞酯酶(Es-1,Es-2和Es-3)的遗传多态性

利用垂直板型聚丙烯酰胺凝胶电泳法,对76只(雌24,雄52)哈尔滨大白兔红细胞酯酶的遗传多态性进行了研究.结果表明,红细胞酯酶Es-1,Es-2和Es-33个位点各有3种不同的表型AA、AB和BB,它们分别受常染色体上的1对等显性基因A、B控制.各位点的A基因频率分别为0.2829、0.5724和0.5329,B基因频率分别为0.7171,0.4276和0.4671.3个位点的杂合度分别为0.4057,0.4895和0.4978,平均杂合度为0.4643.经X~2检验,这3个位点的基因分布均处于Hardy-Weibery平衡状态(P>0.05).     

天津地区部分猪源大肠杆菌的OMP型

从天津各区主要养猪场分离的16株(7种血清型O20、O21、O139、O141、O93、O108、O147)大肠杆菌中提取外膜蛋白(OMP),经SDS-PAGE分析表明,5株O21菌株有2个OMP型(OMP-1和OMP-3型).3株O1s9菌株和3株O20菌株各出现2个OMP型(OMP-1和OMP-2型).2株O141菌株共出现2个OMP型(OMP-2和OMP-3).1株O93菌株、1株O108菌株和1株O147菌株均为OMP-1型.结果表明,相同血清型的菌株可以有不同的OMP型,不同血清型的菌株可以有相同的OMP型.     

Isolation and identification of bubaline herpesvirus 1 (BuHV-1) from latently infected water buffalo (Bubalus bubalis) from Iran

In order to isolate buffaloes herpesvirus 1 (BuHV-1) from latently infected water buffalo (Bubalus bubalis), 16 buffalo heifers were selected from a herd. At first, animals were bled and their sera were tested by virus neutralization (VN) test, using bovine herpesvirus 1 (BoHV-1). According to the results of VN test and dexamethasone injection (0.1 mg/kg BW) for 5 consecutive days, the examined buffaloes were divided into 4 groups. Vaginal and nasal swabs were daily collected from all buffaloes from day 0 to 10 days later. Based on the cytopathic effects in cell culture, a herpesvirus was isolated only from nasal swabs of three seropositive buffaloes which they had received dexamethasone. The nasal swabs of these three buffaloes were also positive in PCR, using primers specific for ruminant herpesviruses gD gene. The identity of the isolated viruses was determined according to partial amino acid sequences of gD, deduced from the nucleotide sequences of the PCR products. On the basis of sequence alignment, phylogenetic analysis, and genetic distances, the three buffalo virus isolates were more closely related to BuHV-1 and BoHV-5 than to BoHV-1.

     

PCR和核酸探针检测猪源沙门氏菌四环素耐药基因tetC的研究

对沙门氏菌的四环素耐药性进行了检测，结果表明，沙门氏菌对四环素类抗生素产生了广泛的耐药性，耐药率达100％，对其四环素耐药基因tetC进行了扩增，结果获得以质粒为模板的特异性产物，与药敏试验结果阳性符合率75％，具有较高的检出率。用光生物素对PcR产物进行标记，制备核酸探针，采用菌落原位杂交的方法对沙门氏菌进行检测，结果表明13株阳性、3株阴性，杂交结果与PCR结果阳性符合率为93．75％，具有较高的特异性。通过条件的优化，建立的四环素耐药基因tetC的PCR和核酸探针检测技术，为四环素耐药性的分子流行病学监测提供了有效的途径。     

Detection of Novel Reassortant Influenza A (H3N2) and H1N1 2009 Pandemic Viruses in Swine in Hanoi,Vietnam

From May to September 2013, monthly samples were collected from swine in a Vietnamese slaughterhouse for influenza virus isolation and serological testing. A(H1N1)pdm09 viruses and a novel H3N2 originating from reassortment between A(H1N1)pdm09 and novel viruses of the North American triple reassortant lineage were isolated. Serological results showed low seroprevalence for the novel H3N2 virus and higher seroprevalence for A(H1N1)pdm09 viruses. In addition, serology suggested that other swine influenza viruses are also circulating in Vietnamese swine.     

犬弓首蛔虫雄虫cDNA文库构建及EST分析

为构建T.canis雄虫cDNA文库,采用Trizol法提取T.canis雄虫的总RNA,合成cDNA,连接到λTripEx2载体上,通过包装蛋白对连接产物的包装,接种到大肠杆菌XL-1-Blue中进行原始文库和扩增文库的滴度测定.经质量鉴定表明:初始文库的滴度为5.25×106 pfu·mL-1,扩增后文库的滴度为6.90×109 pfu ·mL-1.文库的插入片段大小在500～2 000 bp,平均片段大小为1 000 bp,重组率为99.47％.所有指标均显示已成功构建了T.canis雄虫的cDNA文库.利用该文库获得了189条5'有效表达序列标签(EST).对ESTs拼接后代表了101个Unigenes,含有27个Contigs和74个Singletons.其Unigenes在GenBank中的序列号为HO348195～HO348295.同源性分析检索到有56个Unigenes与已知基因同源,其中具有已知或推测功能的基因有40个,未知功能基因有16个,未比对上的基因45个.未比对上的基因与NR数据库中的蛋白序列没有任何意义的匹配,为研究中发现的新基因.这些结果为进一步开展犬弓首蛔虫功能基因及分子机制研究奠定了基础.