Comparison Analysis of Immune Related Gene of Orf Virus in Sheep and Camels

The purpose of this study was to investigate the variation of Orf virus (ORFV) immune related genes after infection with different species.The ORFV genomes of sheep and camel were extracted and named ORFV-Y and ORFV-LT,respectively.Based on ORFV genome sequence published in GenBank (accession No.:KF234407.1),three pairs of specific primers were designed and synthesized to amplify the B2L,F1L and VIR gene fragments of ORFV-Y and ORFV-LT,respectively,and the amplified fragments were cloned into pMD19-T vector,transformed into E.coli DH5α competent cells.The recombinant plasmid was identified,and positive clones were selected for sequencing,DNAStar software was used to analyze the homology,amino acid sequence and phylogenetic tree of 13 ORFV genome sequences published on NCBI.The results showed that the nucleotide homology of B2L,F1L and VIR genes were 92.8% to 99.2%,95.7% to 99.5% and 77.6% to 100%,respectively.After comparing the amino acid sequence between the two genomes and the reference sequence,it was found that there were obvious differences in the immune related genes between the two genomes,and F1L gene had some rules to follow.The phylogenetic analysis of B2L,F1L and VIR genes showed that ORFV-Y was closely related to the Chinese Fujian goat strain,while ORFV-LT was far from the reference strains,and was a separate branch.The results showed that ORFV had obvious difference in immune related genes between sheep and camels,it provided a reference basis for further research on the changes of ORFV gene sequences in different species and the development of vaccines for different species in the future.     

西双版纳橡胶盔蚧天敌调查

通过对西双版纳州各大国有农场(景洪、勐养、勐捧、勐腊、橄榄坝、东风)、民营胶园(三达山、嘎栋、嘎洒、勐罕)及西双版纳热带花卉园内幼林地的调查,初步了解到橡胶盔蚧的天敌31种,其中云南省天敌新记录9种,分为2个纲、7个目、11个科,其中优势种为普通草蛉、川上座壳孢和寄生蜂,分别占天敌总量的26.43%、32.35%和27.48%。比较有利用前景的为普通草蛉和寄生蜂。     

Evaluation of a competitive ELISA for antibody detection against avian influenza virus

Active serologic surveillance is necessary to control the spread of the avian influenza virus (AIV). In this study, we evaluated a commercially-available cELISA in terms of its ability to detect AIV antibodies in the sera of 3,358 animals from twelve species. cELISA detected antibodies against reference H1- through H15-subtype AIV strains without cross reactivity. Furthermore, the cELISA was able to detect antibodies produced following a challenge of the AIV H9N2 subtype in chickens, or following vaccination of the AIV H9 or H5 subtypes in chickens, ducks and geese. Next, we tested the sensitivity and specificity of the cELISA with sera from twelve different animal species, and compared these results with those obtained by the hemagglutination-inhibition (HI) test, the "gold standard" in AIV sera surveillance, a second commercially-available cELISA (IZS ELISA), or the agar gel precipitation (AGP) test. Compared with the HI test, the sensitivities and specificities of cELISA were 95% and 96% in chicken, 86% and 88% in duck, 97% and 100% in turkey, 100% and 87% in goose, and 91% and 97% in swine, respectively. The sensitivities and specificities of the cELISA in this study were higher than those of IZS ELISA for the duck, turkey, goose, and grey partridge sera samples. The results of AGP test against duck and turkey sera also showed significant correlation with the results of cELISA (R-value >0.9). In terms of flock sensitivity, the cELISA correlated better with the HI test than with commercially-available indirect ELISAs, with 100% flock sensitivity.     

饲料中大豆卵磷脂/鱼油比例对斜带石斑鱼稚鱼生长发育及肠道组织的影响

为探讨饲料中不同大豆卵磷脂/鱼油比例对斜带石斑鱼稚鱼成活率、生长行为、饲料利用以及肠道组织学的影响,本实验设计了4种添加不同大豆卵磷脂/鱼油比率的微颗粒饲料：0%/22%（饲料1）,5%/14%（饲料2）,10%/8%（饲料3）以及15%/7%（饲料4）,饲喂斜带石斑鱼稚鱼。每种实验饲料均设3个平行。试验持续40d。结果表明,饲料中不同大豆卵磷脂/鱼油比例对斜带石斑鱼稚鱼生长以及成活率有显著影响（P≤0.05）。饲料3和饲料4的增重分别为：（449.6±11.4）g和（588.3±61.7）g,显著高于饲料1的数值（185.0±2.5）（P≤0.05）。饲料1鱼体脂肪含量为：（154.7±12.8）g·kg-1,显著低于其他饲料组（P≤0.05）,但其鱼体蛋白含量为（728.3±25.5）g·kg-1,显著高于其他饲料组。饲料效率、肥满度、特定生长率、摄食量和蛋白质效率以及终末体重中,4种饲料之间无显著性差异。随饲料卵磷脂/鱼油比例升高,石斑鱼稚鱼肠道脂滴生成数量出现下降,且在饲料4中未观察到肠道脂滴。研究表明斜带石斑鱼稚鱼生长发育阶段需要较高的饲料卵磷脂/鱼油比例（如：15%/7%）。     

One year's study of dynamic and evolution of types I and II PRRSV in a swine farm

This study was to investigate dynamic and evolution of PRRSV in a seed-stock farm by monitoring PRRSV status from 11 June 2009 to 4 August 2010. For laboratory test, around 18-24 umbilical cords from farrowed sows and 5-95 sera from nursery and grow/finish pigs were submitted around every 2 weeks interval during the study. The submitted samples were tested for PRRSV using IDEXX PRRS 2XR ELISA kit, RT-nested PCR. The PRRSV-positive samples were further sequences based on ORF5 and analyzed using MEGA 3.1 program and Beast 1.5.4 package. The surveyed farm was first infected with type II PRRSV but it was infected newly with type I PRRSV of unknown origin, showing rapid substitution to type I PRRSV as a dominant strain in 2 weeks. The type I PRRSV was first detected from umbilical cord of a farrowed sow in 12 January 2010, and secondly from nursery pigs in 26 January 2010. Although sudden increase of mean S/P ratio was found in grow/finish pigs around 2 months earlier than first type I PRRSV detection, no type I PRRSV viremia was found. Thirty three ORF5 full sequences from 14 type II to 19 type I PRRSVs were obtained chronologically in this farm and the genetic characteristics and evolution rates of those sequences were analyzed. The substitution rates (/site/day) of two types were 4.03×10(-5) (type I), 3.09×10(-5) (type II), respectively, which was more frequent than previous reports. The calculated divergence time of type I PRRSV was consistent with the time when the sudden elevation of serum IgG in grow/finish barn was first observed. This study provided fundamental data for type I PRRSV dynamic in a previously type II PRRSV-infected farm and suggested grow/finisher barn could be a primary site for PRRSV introduction.     

ADP-核糖基化调控精子形成的研究进展

ADP-核糖基化是由ADP-核糖基转移酶((ADP-ribosyl) transferases,ARTs)催化的一种蛋白质翻译后修饰,分为聚-ADP-核糖基化和单-ADP-核糖基化,在着丝点和纺锤体组装、DNA损伤修复及精子染色质重塑等多个精子发生生物学事件中发挥重要调控作用.部分ARTs基因缺失使精子形成异常,导致精...     

禽流感病毒重组核蛋白ELISA诊断技术的研究

用表达禽流感病毒（ＡＩＶ）核蛋白基因的杆状病毒感染Ｓｆ９昆虫细胞。以其表达产物制备抗原,建立了以杆状病毒系统表达的ＡＩＶ核蛋白为抗原的禽流感间接酶联免疫吸附试验诊断技术（ｒＮＰ－ＥＬＩＳＡ）。其抗原最适包被量为０．６μｇ／孔,等检血清最适稀释度为１：２００,酶标抗体使用浓度为１：１０００。根据对１４０份ＳＰＦ鸡血清检测结果的统计分析,确定其判定标准为ＯＤ均为阴性;检测Ａ型ＡＩＶ１５个不同亚型（Ｈ１￣Ｈ１５）毒株的特异性血清均为阳性;对人工接种ＡＩＶ的ＳＰＦ鸡第３天即能检出抗体,到第１６２天试验结束时检测仍为阳性。其批内和批间重复试验的变异系数分别在２．９％￣７．２％和３．４％￣９．８％之间。对３１３８份鸡血清进行监测,ｒＮＰ－ＥＬＩＳＡ与全病毒间接ＥＬＩＳＡ与全病毒间接ＥＬＩＳＡ（ＡＩＶ－ＥＬＩＳＡ）、琼脂扩散     

仪器法测定饲料混合均匀度

本研究应用饲料混合均匀度仪测定饲料混合均匀度。考察了仪器的准确性、精确度和稳定性,并将总离子电极仪器法与氯离子选择电极仪器法、邻菲罗啉比色法进行了比较。结果表明:仪器性能稳定、回收率100%、变异系数(CV)小于1.0%。总离子电极仪器法与氯离子选择电极仪器法或邻菲罗啉法无显著差异,具有应用范围广、操作简便、测定快速、结果准确、自动计算等特点。     

CYSTIC ENDOMETRIAL HYPERPLASIA AND BACTERIAL ENDOMETRITIS ASSOCIATED WITH AN INTRAUTERINE FOREIGN BODY IN A GUINEA PIG WITH OVARIAN CYSTIC DISEASE

A 3-year-old, 920 g intact female guinea pig presented with a 4-month history of nonpruritic hair loss on the ventral abdomen. The physical examination revealed flank and ventral abdominal alopecia, mucoid vulvar discharge, and abdominal distension. Bilateral rounded masses just caudal to the kidneys and structures consistent with enlarged uterine horns were detected on abdominal palpation. Abdominal ultrasound revealed bilateral ovarian cysts, thickened uterine horns, and multiple circular hypoechoic and anechoic structures in the uterine wall. The patient underwent ovariohysterectomy. Gross examination of the uterus revealed a piece of hay in the left uterine horn. A final diagnosis was hormonally active ovarian follicular cysts, cystic endometrial hyperplasia, and purulent bacterial endometritis caused by Escherichia coli, Fusobacterium nucleatum, and Arthrobacter spp. Cystic endometrial hyperplasia is infrequently reported in guinea pigs, and this report describes an associated bacterial endometritis and uterine foreign body with concurrent ovarian cysts.     

右美托咪定抑制NOX4缓解急性应激致大鼠肾损伤

本研究拟探索右美托咪定（dexmedetomidine,DEX）对大鼠急性应激致肾损伤的保护作用,并从氧化应激的角度探索DEX对大鼠肾的保护通路。本研究使用了急性束缚应激模型,其中,大鼠被迫游泳15 min,并束缚3 h。本试验采用生化检测、组织病理学切片观察以评估肾功能,然后测定了氧化应激以及氧化应激的相关通路蛋白。旷场试验证实成功建立了急性应激模型。急性应激引起的肾损伤增加了NOX4,降低了Nrf2/HO-1/NQO1表达水平。DEX可降低NOX4表达,同时升高Nrf2/HO-1/NQO1的表达水平。DEX治疗组与急性应激组相比的肾生化结果明显恢复正常,病理切片观察损伤显著降低。试验结果表明,DEX治疗急性应激可影响NOX4/Nrf2/HO-1/NQO1信号通路,并抑制氧化应激。因此,DEX对急性应激引起的肾损伤具有保护作用,并在应激综合征中具有潜在的临床应用。