结核杆菌多位点环介导等温扩增检测方法的建立

为建立一种快速、高敏感、高特异、鉴别结核分枝杆菌和牛分枝杆菌的方法,本研究根据大多数致病性结核分枝杆菌共有序列esat-6,结核分枝杆菌和牛分枝杆菌gyrB共有特异性序列,以及结核分枝杆菌种特异序列mtp40设计6对特异性引物对痰液中的结核分枝杆菌进行检测,并与鉴别培养基分离培养结果以及常规PCR鉴定结果进行比较。实验结果表明,建立的LAMP检测方法具有很高的特异性。可区分致病性结核分枝杆菌和非致病结核分枝杆菌,也可鉴别结核分枝杆菌和牛分枝杆菌。LAMP检测技术的灵敏度比经典PCR技术高100倍左右,可检测到7拷贝/反应。另外,用3种方法同时检测样品发现,LAMP与细菌培养的符合率为90.91%,LAMP与常规PCR检测结果的符合率为100%。LAMP检测方法可以在流行病学调查及现场快速诊断方面广泛应用,并为临床治疗提供依据。     

恒河猴肾上腺皮质的显微与超微结构

应用光镜和透射电镜观察，恒河猴肾上腺皮质可分为球状带、束状带和网状带。其特点是：（1）被膜较发达，有时可伸入球状带将其细胞分隔为团块。（2）球状带有时呈指状指入束状带和网状带，其内可见嗜铬细胞；球状带细胞可分为明、暗2种类型，他们排列成团球状、滤泡状、索状和不规则形。（3）网状带较发达，有的可分为内、外2个区；有时网状带呈刺状伸入束状带；有时网状带细胞可见于被膜下或髓质内。（4）有时部分被膜连同皮质一起呈指状伸入髓质内。（5）皮质细胞均含丰富的溶酶体、线粒体和内质网，但不同部位的细胞所含的脂滴在数量、大小、形态和电子密度等方面存在差异。     

J亚群禽白血病病毒与禽网状内皮增生症病毒共感染对肉鸡生长和免疫功能的抑制作用

1日龄网鸡感染J亚群禽白血病病毒(ALV-J)以及共感染禽网状内皮增生症病毒(REV)后,肉鸡生长发育明显受阻,体重增重明显下降(P＜0.05),法氏囊、胸腺明显萎缩(P＜0.05),在用新城疫疫苗免疫后,感染组血清中新城疫抗体效价显著低于对照组(P＜0.05);在ALV-J和REV共感染后,这种抑制作用更为明显(P＜0.01).ALV J单独感染后,鸡对传染性法氏囊病病毒(IBDV)弱毒疫苗免疫后的抗体反应与对照组没有明显差别,但ALV-J与REV的共感染可明显延缓鸡对IBDV弱毒疫苗免疫的抗体反应.     

Effects of Feeding Different Dietary Crude Protein and Digestible Energy to Yili Mares during Late Lactation Stage on Growth Development and Blood Biochemical Parameters of Foals

In this paper, thirty Yili mares aged 11 to 12 years with weighing 410 kg ± 30 kg, parity of 4 to 5 in third lactation month were selected, and were randomly divided into six groups, each group with five repeats.Two factors quadratic regression orthogonal rotational combination design was adopted to evaluate the effects of feeding different dietary CP and DE to mares on growth development and blood biochemical parameters of foals.Daily DE and CP feeding levels of the 6 groups were 92 MJ/d and 1.20 kg/d, 125 MJ/d and 1.20 kg/d, 92 MJ/d and 1.55 kg/d, 107 MJ/d and 1.35 kg/d, 125 MJ/d and 1.44 kg/d, 115 MJ/d and 1.55 kg/d, respectively.The results showed that body weight, body height, chest circumference and cannon circumference of foals of four and five months were not significant affected by maternal protein and energy intake (P>0.05).As compared with ration contained high DE (125 MJ/d) and low CP (1.20 kg/d), feeding with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) improved body length of foals of four and five months significantly (P<0.05).From the blood biochemical parameters, such as total protein, albumin, triglyceride, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, urea nitrogen, creatinine, alanine aminotransferase, there were no significant differences among all groups (P>0.05).Mares fed with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) obviously improved the aspartate aminotransferase and uric acid level.In summary, Yili mares during lactation late stage fed with 1.35 kg/d CP and 107 MJ/d DE daily could significantly improve body length development of foals, and foals nitrogen metabolism was altered by maternal dietary nutrition through increasing blood aspartate aminotransferase activity.     

Complete Mitochondrial DNA D-Loop Sequence and Genetic Diversity of Wuzhishan Pig

In order to research the genetic diversity and phylogenies of Wuzhishan pig, 56 complete mtDNA D-Loop were analyzed.55 sequences were amplified by polymerase chain reaction (PCR) and 1 sequence was downloaded from GenBank.The results showed that repetitive sequence "TTATAAAACAC" appeared in the conservative regions of 21 mtDNA D-Loop sequences.13 haplotypes and 14 variable sites were observed in 56 sequences.Haplotype diversity and nucleotide diversity (Hd =0.838 and Pi=0.00334) indicated that Wuzhishan pig had high genetic diversity.Two branches in the phylogenetic tree and Network analysis diagram indicated that there were two maternal origins in Wuzhishan pig.The phylogenetic tree of Wuzhishan pig and other 23 species showed that Wuzhishan pig had close genetic relationship with the pig breeds in the south region of the Yangtze river.     

Bioinformatics Analysis of Bone Morphogenetic Protein 4 (BMP4) of Sheep

The assay was aimed to explore the biological characteristics of bone morphogenetic protein 4 (BMP4) of sheep,NCBI,DNAMAN DNAStar,TMHMM Server v.2.0,PsortⅡ,SignalP various bioinformatical softwares were used to speculate the physical and chemical properties,hydrophobic property,phosphorylation site,conservative structure domain,protein secondary structure of BMP4 protein.Also,the three-dimensional structure was forecasted with the SWISS-MODEL Workspace software.The results indicated that the BMP4 of sheep had high homologies with the BMP4 of various species.The encoded protein was a hydrophilic protein which was unstable.There was no transmembrane regions and it was likely to be located in the nucleus.What was more,there was signal peptide and eighteen phosphorylation sites.Through the forecast of functional domains,the protein had two functional domains,including the transforming growth factor-beta (TGF-beta) superfamily and TGF-beta propeptide superfamily.The result was consistent with the function of BMP4 gene family,it also demonstrated that BMP4 was a growth factor and it had the function of signal transduction.The amino acid homology between the predicted 3D structure of protein and template 3bmp.1.A was 88.29%.The bioinformatics analysis of BMP4 gene could provide reference for the further study in practice.     

Study on Biological Characteristics and Drug Sensitivity of Haemophilus parasuis Isolated in Sichuan Province

The study was aimed to investigate the biological characsteristics and drug sensitive tests of Haemophilus parasuis isolated in Sichuan province.This paper reported the biological characteristics,16S rRNA gene PCR and drug sensitive test of the isolates by technique of microbiological and molecular biology.The results showed that the isolates had micro anaerobic and could produce satellite phenomenon,the isolates were just same in biochemical characteristics with field isolates in other area.All the isolates amplified 821 bp 16S rRNA gene.And drug sensitive tests showed that the isolates were highly sensitive to cefradine,ampicillin,cefradine,amoxicillin,cefaclor,ofloxacin,cefotaxime and furadantin,the sensitive rate was 75.61% to 87.80%,but the isolates were high resistance to lincomycin,carbenicillin,enrofloxacin,amikacin and sulfamethoxazole,and multiple-drug resistance was found among the isolates,the number of bacterial strains resistant to antibiotics were more between five and eight,the resisitance strains were 21.95% to 30.49%,7 isolates were even resistant to 13 antibiotics.The results showed the phenomenon of drug resistance of the isolates was very serious.     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

Study on the Nutritional-ecologic Environment of Fe,Cu,Mn,Zn and Se in Yili Summer Pasture of Xinjiang (Ⅰ)

This experiment objective was to study the Fe, Cu, Mn, Zn, Se five kinds of trace element contents, distribution characteristics and mutual relations in the soil-grass-water (stream) in Yili pastoral area of Xinjiang, to evaluate the trace elements nutritional-ecologic environment of Yili summer pasture in Xinjiang.At the altitudinal belt of 1400 to 2999 m of the summer pasture of the Karajon grassland (South slope) and Tangbula grassland (North slope) of Yili, Xinjiang, the samples of soil (n=75), herbage (n=75), stream water (n=75) were collected according to the altitude, to determine their contents of Fe, Cu, Mn, Zn and Se for evaluating the nutritional-ecologic environment of trace elements of Yili summer pasture.The results showed that the contents of Fe, Cu, Mn, Zn in the soil of summer pasture on average were 15 418.4, 32.6, 474.5 and 115.8 mg/kg, respectively;And 446.0, 7.8, 85.7 and 41.9 mg/kg, respectively in forage;The correlation (R²) of the contents of Fe, Cu, Mn and Zn between soil and forage were 0.43, 0.82, 0.47 and 0.72, respectively;The changes of Fe, Cu, Mn and Zn contents in the soil and forage were not significantly related with the altitude.There were no the 4 elements mentioned above in river streams, there was no Se in the soil and forage, and there was infinitesimal of Se in the river streams.It was concluded that in Yili summer pasture there were excessive Fe, suitable Zn and Mn, insufficient Cu, and more insufficient Se.     

扁穗雀麦种质资源形态和农艺性状变异的初步分析

检测种质资源的多样性对于利用和有效管理种质非常重要。本研究对87份世界来源的扁穗雀麦种质利用常用的9个形态-农艺性状进行了统一田间评价。结果表明,所有采用的性状变异系数均表现出高度的变异,尤其以单株干物质产量、分蘖数和叶宽的变异幅度最大。反映株型大小的各性状之间呈显著相关,茎粗和第一节间长与其余8个性状之间存在显著相关(P<0.01),旗叶宽与倒二叶宽的正相关性最强(r=0.912,P<0.01)。主成分分析(PCA)表明前2个主成分可以解释总变异的74%。反映株型大小的性状以及分蘖数可能是扁穗雀麦种质形态变异的主要来源。基于欧氏距离的UPGMA聚类分析将供试材料分成3个主要的类群,第一类群具有最大的株型表现,茎干粗壮,旗叶和倒二叶宽大,分蘖数中等;第二类群具有中等的株型;第三类群具有较小的株型,叶片短而窄,但分蘖数较多。聚类分析很好地支持了前2个主成分分值的二维散点图的结果。总之,本研究表明扁穗雀麦是一种在西南平原和丘陵地区适应性极强的禾草,其表型多样性对于品种选育和资源收集具有重要的参考价值。