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.     

施肥量对泰兴香荷芋生长及产量效益的影响

通过对泰兴香荷芋施肥量研究,考查了各处理主要生育期的苗情与长势、成熟期子孙芋产量及商品芋产值效益。结果显示:基施腐熟脱水猪粪15t·hm-2,苗期追施(3.1叶)脱水猪粪15t·hm-2+13-10-18高钾复合肥675kg·hm-2的处理子孙芋产量最高为23 421.90kg·hm-2,商品芋效益为88 506.60元·hm-2;基施有机肥7 500kg·hm-2+苗期追施(3.1叶)有机肥7 500kg·hm-2+13-10-18高钾复合肥450kg·hm-2+膨大肥(8.1叶)13-10-18高钾复合肥900kg·hm-2的处理产量相对较高。     

黔育1号菊苣与紫花苜蓿、鸭茅间作栽培效果研究

依据贵州省松桃县的气候特点和牧草种植需要,用黔育1号菊苣与北林202紫花苜蓿、黔草4号鸭茅分别进行间作试验,从产量、营养品质、生产效益等方面对间作复合群体与3种牧草单播进行比较分析,探讨黔育1号菊苣与不同牧草间作种植模式的增产效益。结果表明,在贵州松桃地区,黔育1号菊苣+北林202紫花苜蓿间作,年均鲜草产量可达87.27 t/hm²,比紫花苜蓿单播产量增加36.17%,生产效益提高20.77%,粗蛋白含量达26.83%。该研究结果可为松桃县种草养畜产业的发展提供技术支撑。     

不同厂家阿胶及动物皮胶中四种阴离子含量的测定

通过离子交换色谱法,建立不同厂家阿胶及动物皮胶中Cl~-、SO_4~(2-)、NO_3~-、PO_4~(3-)4种阴离子的测定方法。结果表明,4种阴离子分离效果良好,Cl~-、SO_4~(2-)、NO_3~-、PO_4~(3-)分别在0.000 922～0.023 1、0.001 09～0.027 3、0.000 167～0.004 18、0.000 314～0.007 86 mg/mL浓度范围内线性关系良好,平均加样回收率在98.7%～101.0%。该测定方法结果准确、灵敏、可靠,重现性较好,可用于阿胶及动物皮胶中4种阴离子含量的测定。     

Infections Investigation of ALV and REV in Guangxi Local Poultry Breeds and Sequence Analysis of gp85 Gene of ALV Isolates

In order to investigate the infection status of avian leukosis virus (ALV) and avian reticuloendotheliosis virus (REV) in the major local breeds of Qinzhou,Guangxi,totally 953 samples of egg white,cloaca swab and serum of Ma duck,Shitou goose,Tiejiao-Ma chicken,turkey and pigeon were collected from the representing flocks and detected by the commercial ELISA kits.ALV was isolated for the ALV p27 positive samples by culturing on DF-1 cells,and gp85 gene was sequenced.The results showed that the detections of ALV were negative in the samples except those of Tiejiao-Ma chicken,while REV antibody was found positive in Ma duck,Tiejiao-Ma chicken and turkey.The nucleotide sequences of gp85 gene of two isolates shared 94.5% identity with each other,and shared 86.9% to 94.9% with reference strains.The amino acid sequences of gp85 gene of two isolates shared 91.5% identity with each other,and shared 84.0% to 91.6% with reference strains.There were many variable sites in the hyper variable region hr1 and hr2,and the vr2 and vr3 variable regions were relatively conservative.Phylogenetic tree analysis showed that the two isolates shared the highest homology with SCAU11-XG strain.     

Cloning and Sequence Analysis of Buffalo Keap1 Gene and Investigation of Its Expression Pattern in Different Tissues

In this study,the CDS sequence of buffalo Keap1 gene was cloned and analyzed,then its expression pattern in different tissues was also investigated.A pair of primers of buffalo Keap1 gene was designed based on the nucleotide sequence of Bos taurus Keap1 gene from GenBank,and then the buffalo Keap1 gene was amplified.Using the bioinformation techniques,the gene sequence and the protein structure were analyzed.The expression level of Keap1 gene in different tissues were detected with Real-time quantitative PCR.The results showed that the length of buffalo Keap1 gene coding sequence was 1 875 bp and encoded 624 amino acids.The multiple sequence alignment results showed that buffalo Keap1 gene shared 99%,96%,92% and 90% of similar nucleotide sequence with that of Bos taurus,Ovis aries,Sus scrofa and Homo sapiens,respectively.And the phyogenetic tree also showed the conservatism between several different species.The second structure of buffalo Keap1 protein was predicted as 24 alpha regions,40 beta regions,38 turn regions and 27 coil regions.In addition,the results of Real-time quantitative PCR showed that Keap1 mRNA exists in all seven tissues,but the most abundant expression was in heart and the minimal expression was in liver and spleen.The results provided an foundation for further study of Keap1-Nrf2-ARE signal pathway,for enhancing the ability of antioxidant of buffalo embryo in vitro culture.     

Preliminary Study on NF-κB Signaling Pathway Regulating Brucella Intracellular Survival Molecular Mechanisms

By the infection of Brucella virulent strain and attenuated strain in mice macrophage RAW264.7,the assay was aimed to explore the relationship between NF-κB signaling pathways and Brucella virulent strain and attenuated strain in intracellular survival.Use different MOI Brucella (2308,RB51,16M and M5) to infect mice macrophage RAW264.7,after 0,4,8 and 24 h infected,cracking cell and collecting supernatant,we detected the effect of Brucella on activation of NF-κB signaling pathway by Western blotting.Different concentrations of NF-κB signaling pathway inhibitor were incubated with mice macrophage RAW264.7,with different multiplicities of infection (MOI) of Brucella infecting cells,ELISA kits to detect the expressions of TNF-α,IL-1β and IL-6 cytokine;At the same time,count the number of intracellular bacteria of CFU.The results showed that rough cattle Brucella strains RB51 could strongly activate NF-κB signaling pathway,smooth cattle Brucella strains 2308 was weak in the activation;At the same time,the activation of NF-κB signaling pathway was concentration dependent.When the MOI was 80,infection time was 8 h,NF-κB activation degrees of rough cattle Brucella strains RB51 and smooth cattle Brucella strains 2308 were the strongest,and this pathway was involved in producing TNF-α and IL-6;NF-κB signaling pathway inhibitor BAY11-7082 affected Brucella intracellular survival.So rough cattle Brucella strains RB51 intracellular survival and NF-κB signaling pathway activity were closely related.The results laid the foundation for the further study of Brucella intracellular pathogenesis,also provided scientific basis for the research of new drugs to Brucella,and prevention and treatment of brucellosis.     

汉中市耐寒桉树引种繁殖试验

桉树是目前世界上发展最快的短轮伐期工业原料林树种,但多数不耐寒,适宜在热带和亚热带地区发展。为了扩大其北移引种,从四川省林科院引种耐寒品种开展抗寒、抗逆性研究,初步能在汉中安全越冬。本文以邓恩桉全光喷雾嫩枝扦插试验为例,介绍了耐寒桉树繁殖技术。试验采用4种基质、吲哚丁酸3个处理、1个对照、3个重复,结果表明:扦插不同基质处理以珍珠岩∶腐质土(4∶6)最好,吲哚丁酸不同浓度处理间差异不显著,以浓度1 000mg·L-1最好,平均生根率最高达70.83%。