中国矮马运铁蛋白遗传多态性的初步研究

采用聚丙烯酰胺凝胶电泳对中国矮马运铁蛋白多态性进行了测定。在运铁蛋白位点共发现5种表型，即F2F2，DF2，F2H2，F2O和F2R，由Tf^D,Tf^F2,Tf^F2,Tf^H2,Tf^O和Tf^R等5个等位基因控制，其基因频率分别为0.0312,0.7188,0.0625,0.0312和0.1563；基因的杂合度（H），个体鉴别概率（Dp)和亲仔关系排除概率（PE）分别为0.4530,0.6875和0.2586。     

山丹马运铁蛋白遗传多态性的研究

用聚丙烯酰胺凝胶电泳对山丹马的运铁蛋白进行了检测。结果发现该座位有20种基因型，即DD、DF、DF2、DH2、DO、DR、F1F2、F1F1、F1R、F1H2、F2F2、F2H2、F2O、F2R、H2R、H2H2、H2O、OO、OR、RR，其对应的频率分别为0.062，0.021，0.167，0.062，0.042，0.062，0.021，0.021，0.021，0.021，0.021，0.125，0.042，0.146，0.021，0.021，0.021，0.021，0.021，0.062，由Tf^D、Tf^F1、Tf^F2、Tf^H2、Tf^O、和Tf^R6个等位基因控制，其对应频率分别为0.024、0.062、0.323、0.094、0.083、0.198。单位点基因杂合度（H）为0.789，亲仔关系排除概率（PE）为0.584，个体识别概率为0.912。     

四川白猪血液蛋白多态型的测定

采集 30头四川白猪血样 ,采用聚丙烯酰胺凝胶电泳法 ,测定血清中运铁蛋白 (Tf)、前白蛋白(Pa)、后白蛋白 (Po)、血液结合素 (Hp)的多态型 ,估测其基因型频率和基因频率。测定结果表明 :四川白猪运铁蛋白有AA、AB、AC、BB、BC、CC六种表现型 ,前白蛋白有AA、AB、BB三种表现型 ,后白蛋白有AA、AB、BB三种表现型 ,血液结合素有 0 - 1、1 - 1、1 - 2、2 - 2、2 - 3五种表现型 ,其频率分别为 :0 0 6 6 7、0 2 0 0 0、0 2 6 6 7、0 2 0 0 0、0 2 333、0 0 333、0 4 333、0 4 6 6 7、0 1 0 0 0、0 5 333、0 30 0 0、0 1 6 6 7、0 30 0 0、0 4 6 6 7、0 0 333、0 1 0 0 0、0 1 0 0 0     

安哥拉山羊改良建昌黑山羊生产马海毛横交方案的研究

以安哥拉山羊、建昌黑山羊、一代羊(F1)、二代羊(F2)、三代羊(F3)、二代与三代同质个体横交后代(H)为供试羊只,采用形态遗传标记、生化遗传标记和分子遗传标记确定安哥拉山羊改良建昌黑山羊生产马海毛的横交方案。结果:F2、F3 和H白色毛被分别为93 5%、100%和100%,毛被同质和基本同质个体分别为27 7%、70 2%和65 3%;成年羊产毛量随代数增加而增加,但F3 和H与F2,H 与F3 公、母羊分别对应比较,差异均不显著(P>0 05)。成年羊体重F1 和F2 增加,杂种优势明显,但F3 母羊则不显著(P>0 05),H还有下降(P>0 05)。平均基因杂合度和一致度安哥拉山羊、建昌黑山羊、F3、H分别为0 113 1 和0 886 9, 0 062 1 和0 937 9, 0 110 3 和0 889 7, 0 110 4和0 889 6。安哥拉山羊与F3 和H的遗传距离小,分别为0 003 9和0 004 0,遗传相似系数大,相应为0 996 1、0 996 0。群体内谱带相似系数,安哥拉山羊、F2、F3 和H分别为0 454 0,0 407 6,0 611 3 和0 610 9。且F3 和H均极显著大于安哥拉山羊(P<0 01)。综合研究表明,可在F2 和F3 代中选择理想同质个体进行横交。     

生长猪对去皮豆粕中氮和能量利用效率的研究

采用玉米淀粉 豆粕半纯合型日粮 ,选用 36 6 5± 2 6 4kg体重三元杂交去势公猪 12头 ,按 2× 2交叉试验设计 ,研究 5个去皮豆粕和 3个带皮豆粕的养分消化率、氮平衡和能量平衡。结果表明 :去皮豆粕的营养物质利用率高于带皮豆粕。两者日粮的干物质消化率分别为 94 2 5 %和 92 93 % ,氮消化率分别为 93 0 1%和 91 6 2 % ,氮沉积率分别为6 2 2 0 %和 5 9 0 6 % ,能量消化率分别为 95 49%和 94 44 % ,能量沉积率分别为 93 6 9%和 93 0 3 %。而去皮豆粕和带皮豆粕的消化能分别为 15 0 8MJ kg和 14 85MJ kg,相差 0 2 3MJ kg ,差异显著 (P <0 0 5 ) ;代谢能分别为 14 2 9MJ kg和13 85MJ kg,差异 0 44MJ kg ,存在明显差异的趋势 (P <0 1)。总体看来 ,去皮豆粕的营养价值高于带皮豆粕。通过多元线性回归建立豆粕的常规成分含量与消化能和代谢能的回归方程 ,回归系数R2 分别为 0 946和 0 96 4,差异显著性检验后表明 ,F检验显著 (P <0 0 1)。     

河田鸡与白绒乌骨鸡MC1R基因序列分析

为了比较白绒乌骨鸡和河田鸡的黑素皮质素受体1(melanocortin 1 receptor,MC1R)基因序列的差异,试验以NCBI上的红原鸡MC1R基因序列(AB201630)为参照,进行PCR扩增和基因序列比对,并分析了氨基酸序列与蛋白质结构。结果表明:白绒乌骨鸡和河田鸡MC1R基因的序列长度均为945 bp,河田鸡与红原鸡序列同源性为99. 70%,白绒乌骨鸡与红原鸡序列同源性为99. 20%,而河田鸡与白绒乌骨鸡序列同源性为99. 0%;河田鸡相对于红原鸡有2处突变(T143A和C213R),白绒乌骨鸡相对于红原鸡有6处突变(M71T、E92K、L133P、A137T、C213R和H215P),河田鸡相对于白绒乌骨鸡有6处突变(M71T、E92K、L133P、A137T、A143T和H215P);河田鸡和白绒乌骨鸡MCIR基因的二级结构区中α-螺旋、β-折叠、无规则卷曲所占比例分别为55. 73%、8. 92%、35. 35%和52. 55%、12. 42%、35. 03%,均有7个跨膜区,白绒乌骨鸡和河田鸡的三维结构差异主要发生在跨膜区;在鸡形目动物中,MC1R基因具有较高的保守性。说明白绒乌骨鸡MC1R基因突变导致其编码的氨基酸序列H215P异常,推测鸡体内抑制黑色素在羽毛沉积的表征可能由该突变导致。     

秦川牛运铁蛋白多态性与体尺、体重性状的关系分析

秦川牛运铁蛋白 (Tf)多态性与体尺、体重性状之间的关系分析表明 :( 1 )秦川牛体尺、体重的优势基因为TfD1与TfE ;( 2 )在胸深性状上 ,TfEF型的LSE值极显著地大于其它各基因型值 (P <0 .0 1 ) ,TfD1E、TfD1D2 、TfAA、TfEE、TfD2 E、TfD1D1、TfD2 D2 、TfAD1的LSE值显著地大于TfD2 F值 (P <0 .0 5) ;( 3)其他 8个性状各基因型值间差异不显著 (P >0 .0 5)。     

我国部分地区鸡新城疫病毒流行株遗传变异分析

将近年来从不同地区临床病例中分离鉴定的10株鸡新城疫病毒(NDV)流行株,用RT-PCR方法分别扩增出大小为535 bp的F基因重要功能区,并克隆到pGEM-T载体上,经酶切分析结合核苷酸序列测定而确诊。10株分离株核苷酸序列同源性达到81.9%～99.4%,其中8个分离株F蛋白裂解位点的氨基酸顺序为:112R-R-Q/R-K/R/R/F117,为NDV强毒株,且具有基因Ⅶ型的典型特征,即在101位和121位氨基酸残基分别为K(赖氨酸)和V(缬氨酸);其余2株病毒F蛋白裂解位点的氨基酸顺序为:112G-R-Q-G-R-L117,表明它们为NDV弱毒株,在13位和17位氨基酸分别为M(蛋氨酸)和I(异亮氨酸),属于基因Ⅱ型。根据上述测序结果并参照已发表NDV F基因序列绘制出遗传进化树,遗传进化树显示,上述8个强毒株和台湾95年分离株同源性较高,而2个弱毒株与La Sota株同源性较高。     

抗鸡传染性支气管炎病毒核蛋白单克隆抗体的制备及抗原表位鉴定

为鉴定鸡传染性支气管炎病毒(IBV)抗原表位,本研究将IBV tl/CH/LDT3/03株免疫BALB/c小鼠,将其脾细胞与SP2/0骨髓瘤细胞融合,经克隆筛选,得到两株针对IBV核(N)蛋白的单克隆抗体(MAb)6H3和6F9,其染色体数目分别为90条和102条,MAb亚类鉴定分别属于IgG1和IgG2b,轻链均为κ链。对MAb6H3和6F9的抗原表位进行初步鉴定,将N基因分成8个片段克隆于pGEX-6p-1载体中,转化BL21(DE3)内诱导表达。经western blot分析,MAb6H3表位的初步定位在aa80～aa99,而MAb6F9表位的初步定位在aa64～aa82。而且MAb6F9还能特异性识别其他5株IBV的天然N蛋白,推测它们含有一个相同的B细胞表位。     

大庆市猪戊型肝炎病原基因型分析

猪戊型肝炎病毒(H EV)是一种无囊膜的正链单股R NA病毒,直径为27～32nm,表面粗糙,呈不规则球形。H EV基因组全长大约为7.3kb,具有3个互相重叠的开放阅读框架即O RF1、OR F2和O R F3,并且5'端有27个核苷酸的非编码区,3'末端为非结构区。H EV通过喝了污染水而经粪便-口腔途径传播。最近有人从猪血清中检测到抗H EV抗体,并克隆出戊肝人肝炎病毒基因,因此认为戊型肝炎是一种人畜共患性疾病。本研究使用通用引物对来源于猪粪便中的H EV进行了序列分析,以期确定该地区猪群中流行的H EV的基因型,以便更深入的分析猪源H EV与人源H EV…     

Follicle Deviation in Ovulatory Follicular Waves with One or Two Dominant Follicles in Mares

The follicle and hormone aspects of diameter deviation and development of one dominant (≥28 mm) follicle (1DF) vs two dominant follicles (2DF) were studied in 32 ovulatory follicular waves in mares. Follicles were ranked each day as F1 (largest) to F3. The beginning of deviation was designated day 0 and preceded the first increase in the differences in diameter between F1 and F2 in the 1DF group and between a combination of F1 and F2 vs F3 in the 2DF group. One dominant follicle and 2DF developed in 21 (66%) and 11 (34%) waves, respectively. Double ovulations occurred in only one of the waves with 2DF. In 8/11 waves with 2DF, a second deviation occurred between F1 and F2 on 2.5 ± 0.4 days after the first deviation. On day 0, 1DF and 2DF waves were similar in number of days after ovulation, number of follicles, difference in diameter between F1 and F2, and plasma concentrations of LH, estradiol and immunoreactive inhibin. The interval from maximum FSH concentration to day 0 was longer (p < 0.05) and FSH concentration was lower (p < 0.05) on days -1 to 4 in the 2DF group. The similarities on day 0 in the characteristics of 1DF and 2DF waves despite the differences in the declining portions of the FSH profile indicated that a specific day of the FSH decline or a specific concentration were not factors in initiating deviation. Unlike reported results in heifers, the results in mares did not indicate a hormonal basis for the development of 2DF or two deviations.     

平武黄牛血红蛋白和血清运铁蛋白多态性研究

应用醋酸纤维薄膜和垂直板聚丙烯酰胺凝胶电泳技术对四川平武黄牛地方牛种的血红蛋白（Hb)和血清运铁蛋白（Tf)座位多态性进行了分析。结果表明：平武黄牛Hb座位含有3种基因HbA,HbB和HbC,HbC为优势基因，组成5种表现型，同时发现了2头突变个体（基因型暂定为AX型）；Tf座位有4个等位基因，TfD1和TfE为优势基因，组成10种表现型；与秦川牛，宣汉牛比较，Hb和Tf座位的不同基因及基因型频率都有很大差异。     

Difference in content ratio of components among horse serum transferrin variants

Each of five genetic variants of horse serum transferrin (Tf), D, F, H, O, and R, was separated into two bands by polyacrylamide gel isoelectric focusing (PAGIEF). The more acidic band, termed component a, was more abundant than the other one, termed component b, in all variants. Components a and b of TFO variant were immunologically indistinguishable from each other by double immunodiffusion test. Determination of the content ratio of component a to component b in each variant revealed that the variants were classified into two groups: one group (D, F, and H) had a relatively high ratio within a range from 3.4 to 4.0 and the other group (O and R) had a relatively low ratio of 1.8 to 2.3. From these results and reference data on carbohydrate compositions of components a and b, it was proposed that there is a difference in glycosylation between the two groups.     

Serotypes, virulence genes, intimin types and PFGE profiles of Escherichia coli isolated from piglets with diarrhoea in Slovakia

Two hundred and fifty Escherichia coli isolates from diarrhoeic and healthy piglets were serotyped and tested for the presence of virulence genes for fimbriae, intimin, heat-labile (LT) and heat-stable (STa and STb) enterotoxins, Stx toxins, and enteroaggregative heat-stable 1 (EAST1) enterotoxin by polymerase chain reaction (PCR). Although 220 isolates from diarrhoeic piglets belonged to 43 O serogroups and 77 O:H serotypes, 60% were of one of the 10 serogroups O2, O8, O15, O54, O84, O101, O141, O147, O149 and O157, and 60% belonged to only 10 serotypes (O8:H-, O54:H-, O84:H7, O101:H-, O141:H-, O141:H4, O147:H-, O149:H10, O163:H-, and ONT:H-). PCR showed that 79% of 220 isolates carried genes for at least one of the virulence factors tested. The gene encoding for EAST1 was the most prevalent (65%) followed by those encoding for STb (49%), LT (42%), STa (13%), and Stx2e (4%). Eighty-three (38%) of the 220 E. coli isolates carried the gene for F4 (K88), whereas genes for F18, F5 (K99), F41, F6 (P987), F17, and intimin (eae) were detected in 9%, 3%, 3%, 3%, 1%, and 3%, respectively. Seropathotype O149:H10:F4:LT/STb/EAST1 (70 isolates) was the most common, representing 32% of isolates. Pulsed-field gel electrophoresis (PFGE) analysis with XbaI of 15 O149:H10 representative isolates from diarrhoeic piglets distinguished 14 types. The 15 isolates exhibited a wide variability of distinct restriction patterns though all belonged to the same serotype (O149:H10), and all but one showed identical virulence determinants (F4, LT, STb, and EAST1). Among 30 isolates from healthy piglets only two virulence genes were detected: EAST1 (26%) and eae (17%). In total, 12 isolates were positives for the eae gene: five isolates had intimin beta1, four possessed intimin theta and three showed intimin type xiB. This is believed to be the first study describing the presence of intimin type xiB in E. coli of porcine origin.     

Isolation and characterization of nine bacteriophages that lyse O149 enterotoxigenic Escherichia coli

The goal of this study was to isolate and characterize phages that might be used in prevention and treatment of porcine post-weaning diarrhea due to O149 enterotoxigenic E. coli (ETEC). Serotype O149:H10:F4 was especially targeted because this is the dominant ETEC serotype. Mixtures of 10 strains of O149:H10:F4 ETEC and of 10 O149:H43:F4 ETEC were used as hosts for isolation of phages in sewage from 38 Ontario pig farms. Six phages (GJ1-GJ6) that lysed O149:H10:F4 ETEC and three (GJ7-GJ9) that lysed O149:H43:F4 ETEC were isolated. All phages produced large, clear plaques. All nine phages had necks and contractile tails and therefore belonged to the Myoviridae. Their estimated genome sizes were 48.3-50.7kb and their restriction enzyme fragments suggested that they were closely related. Phages GJ1-GJ6 lysed 99-100% of 85 O149:H10:F4 ETEC, 0-12% of 42 O149:H43:F4 ETEC, 3-35% of 37 non-O149 porcine ETEC, and 6-68% of the 72 strains of the ECOR collection. Phages GJ7-GJ9 lysed 86-98% of the O149:H43:F4 ETEC, 2-53% of the O149:H10:F4 ETEC, and 24-41% of the non-O149 porcine ETEC. Titres of the nine phages were unaffected by exposure for 16h to pH 5-9. Among phages GJ1-GJ6, resistance of O149:H10:F4 ETEC to one phage was generally not accompanied by resistance to other phages. It is concluded that the nine phages are suitable candidates for prophylaxis and therapy of porcine post-weaning diarrhea due to O149 ETEC.     

三种鹿属动物血清淀粉酶同工酶的电泳研究

采用聚丙烯酰胺凝胶电泳法对３４头白唇鹿，５８头马鹿和１１头梅花鹿的血清淀粉酶同工酶进行了比较研究。结果发现：（１）３种鹿的血清淀粉酶都有ＡＭＹ１，ＡＭＹ２和ＡＭＹ３种同工酶，但白唇鹿ＡＭＹ１和ＡＭＹ２同工酶的相对迁移率小于马鹿和梅花鹿；（２）３种鹿的ＡＭＹ１同工酶都有ＡＭＹ１Ｆ和ＡＭＹ１Ｓ两种表型，以ＡＭＹ１Ｆ为优势表型（分别为６４．７１％，６８．９７％和８１．８２％）；（３）白唇鹿的ＡＭＹ２同工酶存在ＡＭＹ２ＡＡ，ＡＭＹ２ＡＢ和ＡＭＹ２ＢＢ３种表型，以ＡＭＹ２ＡＡ为优势表型（５０．００％）；（４）马鹿的ＡＭＹ２同工酶存在ＡＭＹ２ＡＡ，ＡＭＹ２ＡＢ，ＡＭＹ２ＢＢ和ＡＭＹ２Ｏ４种表型，梅花鹿存在ＡＭＹ２ＡＢ和ＡＭＹ２ＢＢ两种表型，以ＡＭＹ２ＡＢ为优势表型（分别为８９．６６％和９０．９１％）。     

Frequency of Shiga toxin-producing Escherichia coli in cattle at a breeding farm and at a slaughterhouse in Japan

To investigate the frequency of Shiga toxin-producing Escherichia coli (STEC) infected calves at a breeding farm and cattle at a slaughterhouse in Tohoku area of Japan, the polymerase chain reaction (PCR) was used for detection of genes for Shiga toxin(s). The fecal samples from a total of 204 calves and 306 cattle were examined. The prevalence rates in calves less than 2 months of age, cattle 2-8 months of age, and adults greater than 1 year of age were 39.4, 78.9, and 40.8%, respectively. Detection frequency of STEC in the fecal specimens from calves aged 0-8 months was not different among the breeds of cattle (Holstein: H, Japanese black cattle: B, and F1: HxB). On the other hand, for calves over 12 months of age, the frequency of STEC in Japanese black cattle and F1 were significantly higher than in Holstein cattle. Serogroups of STEC usually identified in human cases of food poisoning (O157, O26, and O111) were not frequently found in the feces of the cattle.     

日本大耳白兔、新西兰兔血液蛋白多态性的研究

采用垂直板聚丙烯酰胺凝胶电泳法对日本大耳白兔、新西兰兔血液中的血清转铁蛋白 (Tf)、前转铁蛋白 (Prt)、后白蛋白 (Po)、血液结合素 (Hp)、红细胞酯酶 (ES - 1、ES - 2和ES - 3)等 7个蛋白座位进行测定。结果表明 :这 7个座位均呈多态。在多态座位中 ,Tf、Prt、Po、Hp、ES - 1、ES - 2、ES - 3分别出现 3、3、6、3、3和 3种表型 ,受 2、2、3、2、2和 2个共显性等位基因控制。X2 检验表明 ,新西兰兔在这 7个座位上均处于平衡状态 (P >0 .0 5 ) ,日本大耳白兔在Tf、Po、ES -1、ES - 34个座位处于平衡 (P >0 .0 5 ) ,而在Prt、Hp、ES - 2出现不平衡状态 (P <0 .0 5 )。     

中国矮马血液遗传标记的初步研究

采用聚丙烯酰胺凝胶电泳对16匹中国矮马血清中白蛋白（ALB）、酯酶（ES）、α1-B糖蛋白（A1B）和维生素D结合蛋白（GC）进行了检测。结果发现：在4个位点中，白蛋白（ALB）和酯酶（ES）位点呈现多态性，而α1-B糖蛋白（A1B）、维生素D结合蛋白（GC）2个位点均呈现单态性。在白蛋白（ALB）位点发现3个基因型，即AA、AB和BB，由等位基因ALB^A和ALB^B控制，其基因频率分别为0.3125和0.6875；在酯酶（ES）位点发现4个基因型，即FS、II、FF和FI，由等位基因ES^s、ES^1和Es^f控制，其基因频率分别为0.1875,0.281和0.5313。