新型绿色饲料添加剂-螺旋藻

螺旋藻广泛分布在世界各海区及陆地淡、咸水湖中 ,是浮游自养的原核生物 ,属蓝藻门颤藻目颤藻科。藻体由于含藻蓝素而呈绿色 ,显微镜观察呈细丝状螺旋形。全世界已知存在 3 6个种 ,目前国际上工厂化生产的主要是钝顶螺旋藻和极大螺旋藻 2个淡水种类。螺旋藻是目前常用微藻 (小球藻、绿藻、螺旋藻 )中蛋白质含量最高、营养全面、消化吸收及适口性最好的藻种 ,而且其安全性很好 ,至今未发现对动物有任何毒性或致病等副作用。1　螺旋藻的营养特点1 .1　蛋白质含量高 ,氨基酸组成合理螺旋藻含蛋白质高达 5 5 %～ 70 % ,相当于大豆蛋白质的 2倍 ,…     

处理猪场沼液的微藻筛选及其净化效果评价

本研究旨在筛选可高效降解猪场沼液污染物并具有产蛋白和油脂潜力的微藻藻种,探讨其对不同稀释浓度沼液的处理效果。通过对沼液原位富集筛选,分离获得4株微藻,分别命名为CS-1、CS-2、CS-3和CS-4,分析不同微藻的生长及油脂、蛋白质产率。结果表明:藻株CS-3的生物量产率、蛋白产率、油脂产率和细胞内油脂含量均高于其余藻株(P<0.05),因而选取藻株CS-3为目标藻株,经过18S rDNA鉴定属于小球藻属(Chlorella sp.);比较不同微藻对沼液的处理效果,藻株CS-3对硝酸盐氮、总氮、总磷和化学需氧量等水质指标的去除效果最好,去除率分别达到94.5%、90.0%、94.8%和46.1%;在光照强度3000lux、温度25℃、曝气强度2.0L/min的条件下,CS-3对6倍和4倍稀释的猪场沼液处理效果较好,经过10 d处理,总氮的去除率分别为96.4%和94.4%,氨氮的去除率为98.4%和98.2%,总磷的去除率为100%和96.2%。以上表明,小球藻CS-3在沼液处理中具有应用前景,且其对沼液的净化效果受沼液稀释倍数的影响。     

我国东北部分地区牛溶血性曼氏杆菌的分子流行病学研究

为了解牛溶血性曼氏杆菌(Mannheimia haemolytica,Mh)在我国牛群中的流行情况,本研究于2014年～2019年在东北部分地区采集健康牛、患病牛的鼻拭子和肺脏样品进行细菌的检测、分离和鉴定,并对分离株进行遗传进化分析。对牛溶血性曼氏杆菌的检测结果显示,不同年份、不同地区采集样品的总检出率为23.5%(28/119),其中健康牛的检出率为13.46%(7/52),明显低于临床发病或死亡牛样品的检出率31.34%(21/67)。采用特异性分型PCR检测发现,Mh分离株分为A1、A2和A6三个血清型,从健康牛分离的Mh株的主要血清型为A2型,而从发病或死亡牛主要分离的Mh是A1和A6型。遗传进化分析表明,A1和A6型分离株位于同一进化分支,菌株间16S rDNA和lktA基因的序列同源性均为100%;而它们与A2型分离株的亲缘关系较远,16S rDNA和lktA基因的序列同源性分别为99.8%～99.9%和85.8%～86.7%;此外,A1和A6型分离株的lktA同属于lktA1等位基因亚型,而A2型分离株属于lktA2和lktA8亚型,与A1和A6型相比呈现出较强的序列多样性。综上所述,Mh在我国东北部分地区牛群中的存在较为普遍,且A1和A6血清型是牛群中Mh的主要致病血清型。本研究获得的流行病学数据为我国牛群中溶血性曼氏杆菌的病原学以及疫苗研究提供科学依据。     

鸡肾型传染性支气管炎病毒分离株S1基因的序列分析

采取RT-PCR方法对5株传染性支气管炎病毒分离株的S1基因进行序列扩增、测定及分析,应用DNAStar软件将这些分离毒株与中国常用疫苗毒株、国际上其它血清型的代表参考毒株分别进行核苷酸和氨基酸系统发生进化关系分析.核苷酸与氨基酸序列分析结果表明,这5株分离毒株与A2毒株的亲缘关系较近,与疫苗株H120和Ma5的亲缘关系较远.     

鸡肾型传染性支气管炎病毒分离株S1基因的序列分析

采取RT-PCR方法对5株传染性支气管炎病毒分离株的S1基因进行序列扩增、测定及分析,应用DNAStar软件将这些分离毒株与中国常用疫苗毒株、国际上其它血清型的代表参考毒株分别进行核苷酸和氨基酸系统发生进化关系分析。核苷酸与氨基酸序列分析结果表明,这5株分离毒株与A2毒株的亲缘关系较近,与疫苗株H120和Ma5的亲缘关系较远。     

福建省羊传染性脓疱病毒F1L、B2L和VIR基因的遗传变异分析

为探明2014年-2015年在福建省流行的羊传染性脓疱病毒(ORFV)遗传变异情况,对10株ORFV流行毒株的F1L、B2L和VIR基因进行克隆、测序及分析。结果表明,10株ORFV F1L基因之间的核苷酸序列同源性为97.6%~100%,与国内株的核苷酸序列同源性为96.8%~99.7%,与NZ2参考株的核苷酸序列同源性为96.3%~97.1%;同NZ2参考株比较,FJ-YT2014缺失2个氨基酸;10株ORFV B2L基因之间核苷酸序列同源性为97.5%~99.9%,与国内株的核苷酸序列同源性为96.7%~99.5%,与NZ2参考株的核苷酸序列同源性为96.7%~97.7%;10株ORFV VIR基因之间的核苷酸序列同源性为95.8%~99.5%,与国内株的核苷酸序列同源性为94.6%~99.6%,与NZ2参考株的核苷酸序列同源性为94.6%~96.4%。基于基因核苷酸序列的遗传进化分析表明,10株ORFV F1L基因与福建省分离株、山西株和新疆株亲缘关系较近;10株ORFV B2L基因与新疆、山西、德国毒株亲缘关系较近;10株ORFV VIR基因与台湾、新疆株亲缘关系较近。结果提示,当前福建省流行的ORFV F1L、B2L和VIR基因尚未出现明显变异,但是其F1L、B2L和VIR基因核苷酸序列之间普遍存在异质性。     

新疆塔城地区马梨形虫感染的分子生物学诊断及分析

采用常规血涂片镜检和PCR技术对新疆塔城地区疑似梨形虫感染的病马进行检测。通过18S rRNA基因通用引物扩增出目的基因,克隆测序后,与GenBank公布的梨形虫18S rDNA参考序列对比分析同源性,并构建系统进化树,确定梨形虫的种属。血涂片鉴定结果显示,红细胞中发现圆环形虫体,具有马梨形虫的主要特征。分子生物学鉴定发现,18S rDNA序列与马泰勒虫美国分离株核苷酸序列同源性为98.3%;遗传进化分析发现,该虫株与已知的泰勒属在同一大枝上,与马泰勒虫美国分离株、马泰勒虫西班牙分离株有较近的亲缘关系。     

水产养殖用复合微生物粉剂中菌种的分子生物学鉴定

对两种市售水产养殖用复合微生物粉剂中微生物菌种进行鉴定。利用平板分离技术对粉剂中的微生物进行分离纯化,进行了形态学观察、16S rDNA和26S rDNA序列分析,并构建系统发育树。结果表明,两种微生物粉剂中菌群结构存在一定差异,S1粉剂中鉴定出5株芽孢杆菌和2株酵母菌,分别为枯草芽孢杆菌、地衣芽孢杆菌或索诺拉沙漠芽孢杆菌、葡萄牙棒孢酵母和库德里阿兹威毕赤酵母;S2粉剂中鉴定出3株芽孢杆菌、1株放线菌、2株酵母菌和3株肠球菌,分别为解木糖赖氨酸芽孢杆菌、环状芽孢杆菌、地衣芽孢杆菌或索诺拉沙漠芽孢杆菌、芬氏纤维微菌、酿酒酵母、库德里阿兹威毕赤酵母、鹑鸡肠球菌、屎肠球菌或乳酸肠球菌。明确水产养殖用复合微生物粉剂的菌群结构为同类产品的安全性及有效性评估提供理论基础。     

一株H5N3亚型禽流感病毒全基因组序列分析及其对小鼠的感染性研究

为了解H5N3亚型流感病毒的生物学特性,本研究对2017年浙江省分离到的一株H5N3亚型禽流感病毒(AIV)[DK/ZJ/S1368/2017(H5N3)]进行了遗传演化分析及小鼠感染性实验。遗传演化分析结果显示,该株病毒的HA蛋白裂解位点处仅含一个碱性氨基酸,属于低致病性AIV。同时,该病毒的血凝素(HA)基因与H5N7亚型流感病毒亲缘关系较近;聚合酶碱性蛋白2(PB2)基因和核蛋白(NP)基因与H10N7亚型流感病毒亲缘关系较近;碱性聚合酶蛋白1(PB1)基因与H1N1亚型流感病毒亲缘关系较近;酸性聚合酶蛋白(PA)基因与H6N2亚型流感病毒亲缘关系较近;神经氨酸酶(NA)基因与H10N3亚型流感病毒亲缘关系较近;基质蛋白(M)基因与H3N8亚型流感病毒亲缘关系较近;非结构蛋白(NS)基因与H1N1亚型流感病毒亲缘关系较近。表明其基因来源复杂。小鼠感染实验结果显示,该分离株无需提前适应即可以在小鼠肺脏和鼻甲中复制,小鼠感染病毒后无明显临床症状,与对照组相比其体质量变化不明显,表明该病毒对小鼠呈低致病性。本研究通过对该H5N3亚型AIV的生物学特性的分析,发现该病毒基因来源复杂,无需提前适应就可以在小鼠体内复制,具有感染哺乳动物的潜在威胁,提示应当持续加强对H5N3亚型AIV的监测和相关生物学特性的研究工作。     

野生动物源肠杆菌科细菌鉴定及同源性分析

本文主要对2020-2021年间分离自杭州动物园患病动物、死亡动物的肠杆菌科细菌进行鉴定和同源性分析。通过16S rDNA技术鉴定细菌种类、通过MAGA6.0方法对16S rDNA扩增序列构建系统发生树,并结合ERIC-PCR指纹图谱技术来分析分离的肠杆菌科细菌的同源性。结果共分离保存菌株38株,经16S rDNA鉴定,其中克雷伯氏菌4株,埃希氏菌27株,摩氏摩根菌2株,沙门氏菌1株,弗氏柠檬酸杆菌1株,哈夫尼氏菌2株,勒克氏菌1株;对埃希氏菌属构建系统发生树显示,置信度在90%可分为23个亚群,其中E1和E26,E5、E6、E17和E18,从进化关系上看更接近,通过NTSYSpc2.0软件对ERIC-PCR指纹图谱分析,在相似系数为0.87时,埃希氏菌属可分为18个亚群。该研究通过对杭州动物园野生动物源肠杆科细菌鉴定,以及采用ERIC-PCR对埃希氏细菌的同源鉴定,结果表明临床埃希氏菌属细菌感染非同株菌感染引发,这对临床疾病预防和肠杆菌科细菌快速分类有一定的指导意义。     

Evaluation of two monoclonal antibodies for serotyping Haemophilus paragallinarum

Two monoclonal antibodies (MAbs) were evaluated for their ability to serotype 108 isolates of Haemophilus paragallinarum. One MAb (E5C12D10) was raised against a Page serovar A strain and the other (F2E6) against a Page serovar C strain. In both dot blot and hemagglutination-inhibition tests, MAb E5C12D10 recognized the type strains of Page serovar A and Kume serovars A-1, A-2, A-3, and A-4. MAb F2E6 recognized the type strains of Page serovar C and Kume serovars C-1, C-2, and C-3. Neither antibody recognized the type strains of Page serovar B or Kume serovars B-1 and C-4. When evaluated with 97 field isolates in a dot blot test, the MAbs serotyped 81 isolates, which was better than agglutinin typing by the Page scheme (69 isolates serotyped). The field isolates that did not react with the MAbs were either Page serovar B/Kume serovar B-1 (three isolates), Page serovar C/Kume serovar C-4 (12 isolates), or nontypable by either the Page or Kume scheme (one isolate).     

Generation of monoclonal antibodies to porcine interleukin 6 (PIL-6) using the recombinant PIL-6 expressed in Escherichia coli

Porcine interleukin-6 (PIL-6) protein without signal peptide was expressed as a glutathione S-transferase (GST) fusion protein in Escherichia coli. The fusion protein was expressed in an insoluble fraction, however, it was solubilized by refolding procedure using urea. From the solubilized protein, the recombinant PIL-6 (rPIL-6) was purified by a batch method using glutathione sepharose 4B and PreScission protease cleavage. By the B3B1 hybridoma cell proliferation assay, biological activity of the purified rPIL-6 was confirmed. Three monoclonal antibodies (MAbs) named 2B-1, 5A-8 and 4C-3 were generated by using the rPIL-6 as an immunogen. Immunoglobulin isotypes of the MAbs were IgG2a (4C-3) and IgG2b (2B-1 and 5A-8). For the epitope analysis, additive enzyme-linked immunosorbent assay and immunoblot analysis using deletion mutants of PIL-6 were performed. These experiments revealed that the two MAbs (2B-1 and 5A-8) recognize an overlapped epitope and the other (4C-3) recognizes a distinct epitope, and all epitopes reside in the region of aa26-64 of PIL-6.     

Pathogenicity of various adenovirus serotypes in the presence of Escherichia coli in chickens

Ten strains of adenovirus representing 10 serotypes were administered intratracheally to 3-week-old specific-pathogen-free chickens, which also received 2.9 X 10(5) colony-forming units of a pathogenic Escherichia coli intranasally. One group was given only E. coli, and one was retained as an uninoculated control. Gross pathologic alterations post-mortem were minimal and limited to multiple scattered, pale areas in the lungs of an occasional chicken in various groups. Histopathologic changes in the lungs were those of multifocal, interstitial, and occasionally diffuse pneumonia. Moderate to marked interstitial pneumonia was incited by adenovirus strains 75-1A, B-3 A-2, C-2B, and X-11; Ind-C, Stein, Tipton, J-2, and T-8 caused similar but milder lesions. Strains 75-1A, A-2, C-2B, T-8, and X-11 incited moderate to marked multifocal pneumonia; Ind-C, Stein, Tipton, J-2, and B-3 caused mild multifocal pneumonia. In all groups, the pneumonic lesions were more severe 5 days postinoculation than 12 days postinoculation. Bronchiolitis and tracheitis lesions also varied in severity with serotype. A mild hepatitis was seen with serotypes T-8 and 75-1A. Neither the uninoculated control group nor the group inoculated with only E. coli exhibited gross or histopathologic alterations.     

马铃薯杂种优良株系遗传差异的AFLP分析

为明确从‘1867’ב陇薯7号’、‘MB09’ב陇薯7号’和‘MB09’ב陇薯6号’3个马铃薯杂交组合中选育出的17个杂种优良株系的遗传差异程度,用4个亲本材料作对照,利用AFLP分子标记技术对其进行了遗传差异分析。用筛选出的10对AFLP适宜引物进行PCR扩增共得到321个位点,多态性位点277个,多态性位点百分率87.29%。试验建立了各杂种优良株系的AFLP指纹图。21份马铃薯材料间的多态信息含量(PIC)平均为0.5617,Nei’s遗传多样性指数为0.3623,Shannon指数为0.5407。各材料间的平均遗传距离(GD)为0.5281,以GD值0.51为基准,21份材料分为4类:‘1867’、A-10、A-12、A-21和A-29为一类;‘陇薯6号’、‘陇薯7号’、A-14、A-23、A-26、B-13和B-14为一类;‘MB09’、B-1、B-2、B-7、B-15、B-20和C-22为一类;C-2和C-21为一类。该研究可为马铃薯杂交亲本的选择利用和杂种优良新品系的选育提供依据。     

Serotyping of Haemophilus paragallinarum isolates from Mexico by the Kume hemagglutinin scheme

A total of 42 isolates of Haemophilus paragallinarum from Mexico were serotyped by the Kume hemagglutinin scheme. Serovars A-1, A-2, B-1, and C-2 were recognized among 11 (26.2%), 7 (16.6%), 4 (9.5%), and 14 (33.3%) isolates, respectively. A further six isolates (14.3%) showed hemagglutinating activity but could not be classified into any serovar. Commercial vaccines containing Kume serovars A-1, A-2, B-1, and C-2 may provide better protection than those bi- or trivalent infectious coryza vaccines currently used in Mexico.     

Development of an experimental model allowing discrimination between virulent and avirulent isolates of Serpulina (Treponema) hyodysenteriae.

Variation in virulence among different strains of Serpulina hyodysenteriae was studied by oral inoculation of specific pathogen free piglets and CD-1 mice. Piglets infected with serotype 2 reference strain B204 and an untypable field strain LHV-90-9-I had severe diarrhea tainted intermittently with mucus and fresh blood. The piglets inoculated with B169, B8044, B6933, and ACK300-8 reference strains representing serotypes 3, 5, 6, and 7 respectively developed moderate diarrhea. However, reference strains B234 and A-1 of serotypes 1 and 4, respectively, failed to cause any diarrhea. None of the S. hyodysenteriae strains caused diarrhea in mice. The results indicate a great variation in virulence among strains of different serotypes of S. hyodysenteriae. Mice were less susceptible to infection with S. hyodysenteriae.     

Crossbreeding cattle in beef production programmes in Kenya

Data collected on a privately owned ranch located in the Machakos District of Kenya at approximately 2 degrees latitude south of the equator at an elevation varying from 1,675 to 2,000 m were analysed on five breed groups of cows: (1) purebred Boran, (2) 1/2 Charolais-1/2 Boran (1/2 C-1/2 B), (3) 3/4 Boran-1/4 Charolais (3/4 B-1/4 C), (4) 1/2 Ayrshire-1/2 Boran (1/2 A-1/2 B) and (5) 1/2 Santa Gertrudis-1/2 Boran (1/2 SG-1/2 B). The maternal traits evaluated included age at first calving, calving interval, calf weight at weaning and cow productivity index (calf weight weaned annually per cow calving). Mean cow productivity index for all cows was 192 kg; for purebred Boran, 174 kg; for 1/2 C-1/2 B, 200 kg; for 3/4 B-1/4 C, 191 kg; for 1/2 A-1/2 B, 210 kg; and for 1/2 SG-1/2 B, 185 kg. Cow breed groups 1/2 C-1/2 B, 3/4 B-1/4 C, 1/2 A-1/2 B and 1/2 SG-1/2 B exceeded (P less than 0.01) purebred Boran by 14.9, 9.8, 20.7 and 6.3%, respectively, in cow productivity index.     

Protection conferred by bivalent and trivalent infectious coryza bacterins against prevalent serovars of Avibacterium (Haemophilus) paragallinarum in Mexico

The protection and level of hemagglutination-inhibition (HI) antibodies conferred in infectious coryza bivalent- and trivalent-immunized chickens against Avibacterium (Haemophilus) paragallinarum field isolates of the prevalent serovars in Mexico (A-1, A-2, B-1, and C-2) were investigated. The bivalent bacterin (A-1 and C-1) conferred significant protection and increased HI antibodies against isolates of serovars A-1, A-2, and C-2, but not against a serovar B-1 isolate. The trivalent bacterin (A-1, B-1, and C-2) conferred protection and increased HI antibodies against all four of the isolates. The results confirmed that in poultry areas where serovar B-1 is prevalent, the inclusion of this serovar in bacterins is needed to confer protection against infectious coryza caused by A. (H.) paragallinarum isolates of serovar B-1.     

Typing of Haemophilus paragallinarum strains by using enterobacterial repetitive intergenic consensus-based polymerase chain reaction

The enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique was used for fingerprinting of reference strains and Mexican isolates of Haemophilus paragallinarum. A total of nine ERIC patterns were given by the nine serovar reference strains of this bacteria. Two Modesto (C-2) reference strains from different sources showed the same ERIC pattern. Seventeen ERIC patterns were obtained among 29 Mexican isolates included in the study, belonging to serovars prevalent in Mexico (A-1, A-2, B-1, and C-2). Obtained results indicate that the ERIC-PCR technique could be used as a molecular laboratory tool for subtyping of H. paragallinarum.     

白尾地鸦种群、繁殖生态及保护现状

白尾地鸦(Podoces biddulphi)仅分布于新疆塔克拉玛干沙漠沙漠之中,为中国特有物种.目前有关这个种的现状和生态报道较少.自1988年以来,作者在塔克拉玛干沙漠腹地对白尾地鸦种群状况、分布、繁殖生态等进行了系统的研究.并对沙漠化造成白尾地鸦"东扩"进行了分析和讨论.