Immune responses of orange-spotted grouper,Epinephelus coioides,against virus-like particles of betanodavirus produced in Escherichia coli

Betanodaviruses are the causative agents of viral nervous necrosis (VNN), a serious disease of cultured marine fish worldwide. Virus-like particles (VLPs) are one of the good novel vaccine candidates to control this disease. Until now, betanodavirus vaccine studies mainly focused on the humoral immune response and mortality after virus challenge. However, little is known about the activation of genes responsible for cellular and innate immunity by vaccines. In the present study, VLPs of orange-spotted grouper nervous necrosis virus (OGNNV) were produced in prokaryotes and their ability to enter Asian sea bass cells was the same as native virus, suggesting that they possess a similar structure to OGNNV. VLPs immunogenicity was then determined by intramuscularly vaccinating Epinephelus coioides at different concentrations (1.5 or 15 μg g^?1 fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5 μg g^?1 FBW is enough to provoke high titer antibodies (average 3 fold higher than that of negative control) with strong neutralizing antibody titer as early as 1 week post immunization. Furthermore, quantitative PCR analysis revealed that eleven genes associated with humoral, cellular and innate immunities were up-regulated in the liver, spleen and head kidney at 12 h post immunization, correlating with the early antibody response. In conclusion, we demonstrated that VLP vaccination induced humoral immune responses and activated genes associated with cellular and innate immunity against betanodavirus infection in orange-spotted grouper.     

2006—2012年鲁豫冀地区15株猪繁殖与呼吸综合征病毒的分离鉴定及ORF5/Nsp2基因的遗传变异分析

为研究鲁豫冀地区猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)的遗传变异情况,对2006—2012年来自3省区发病猪场的42份样品进行PRRSV分离鉴定,并进行了生物学特性研究和PCR鉴定,结果显示先后分离到15株PRRSV。分别采用RT-PCR扩增其ORF5基因和部分Nsp2基因并测序,与GenBank中68个ORF5序列和40个Nsp2序列的推导氨基酸序列进行比对,遗传变异分析结果表明,15株分离株均属于美洲型毒株,其中13个毒株Nsp2基因推导的氨基酸序列均存在氨基酸的不连续缺失,其ORF5基因推导的氨基酸序列与JXA1株有较高的同源性(95.5%~97.5%);SDDY2007株与疫苗株RespPRRS MLV和VR2332株亲缘关系相近,处于同一个亚群中;而HN25-2009分离株Nsp2基因推导的氨基酸序列有30个氨基酸的特征性缺失,其ORF5基因推导的氨基酸序列的遗传进化分析结果显示该分离株处于VR2332所在亚群(氨基酸同源性97.5%),具有一定特殊性。本试验结果表明,2006—2012年高致病性PRRSV是鲁豫冀地区的优势流行毒株,且存在疫苗毒株,3省区流行毒株间有一定遗传差异,但无明显地域特征。     

基于超平面构建的高光谱异常目标检测方法

高光谱遥感影像具有光谱分辨率高、图谱合一的特点,影像中的地物组成也多种多样,针对图像数据中目标出现概率较小、检测概率较低等问题,本文提出了基于超平面构建的高光谱数据目标异常检测方法。它通过对高光谱数据的结构分析,构建出高维空间中的背景超平面算子,以待测像元在其正交子空间的投影值来表征异常程度的大小。通过OMIS和AVIRIS实测数据验证,并与RX算法相比较,基于超平面构建的高光谱异常检测算法能够在多种背景环境下检测出纯度高、光谱差异大的异常像元,并具有较高的检测率和较低的虚警率,因此具备了有效性和较好的实用性。     

台湾控制禽流感的措施

台湾控制禽流感的措施主要有3个方面,第一,对鸡、鸭、鹅以及候鸟进行禽流感主动和被动的监测,这个工作有6个单位参与,4所大学和2个研究所,即台湾动物健康研究所、台湾动物科技研究所、台湾大学、嘉义大学、中兴大学、屏东大学;第二、注重交流,官员、禽流感专家以及养殖户经常通过会议或讲座进行交流;第三,加强生物安全或扑杀,台湾野鸟协会每年都要采集2 000～3 000个粪便样本来监测候鸟中禽流感病毒的存在情况.     

ELISE测定猪尿、猪肝中克伦特罗残留的研究

ELISA(酶联免疫吸附法)测定猪尿、猪肝中违禁药物克伦特罗残留量影响实验结果表明,德国拜发与北京望尔生产的两种ELISA试剂盒具有操作简单、时间短、成本低、敏感度高等特点。在室温20-22℃条件下,试验制备过程中必须做到控温度、控转速、控流速及洗板过程中应注意事项,这样可大大提高在实际操作过程中检测的准确性。     

放牧对鸭茅能量积累及分配的影响

通过长期放牧试验，探讨了不同放牧强度下亚热带高山草地鸭茅种群能量积累规律及能量在不同构件之间的分配特征。在无放牧的对照草地及采食率分别为55％、65％、80％的放牧草地，鸭茅热值随放牧强度的增加而呈单峰曲线型变化，一定强度范围内的放牧干扰提高了鸭茅热值及能量的积累；根部能量的积累随放牧强度的提高而逐渐减少。在禁牧草地，鸭茅种群能量主要被低密度的大分蘖丛所分摊，随着放牧强度的提高，种群能量则逐渐均摊给高密度的小分蘖丛。     

微生物菌群培养组学在动物医学中的应用

实验室条件下可培养的微生物约占自然界中微生物总数的1%,这限制了人们对99%未知微生物的认识和利用,而研究表明,那些“不可培养的微生物”是可以被开发和利用的,未能被纯培养的微生物才是未知微生物的主体。微生物培养组学探索利用多种培养条件和长时间的培养,结合基质辅助激光解吸电离飞行时间质谱法（MALDI-TOF-MS）和16S核糖体RNA（rRNA）测序可以大规模鉴定各种微生物,同时利用全基因组测序和宏基因组测序手段对未知微生物进行深入分析。本文综述了国内外近年来微生物菌群培养组学在反刍动物胃肠道、禽类盲肠及家畜鼻腔微生物菌群研究中的最新进展,探讨将动物体内菌群培养组学方法应用于动物疾病防治领域的可行性。作为一个新兴的研究方法,尽管该培养组学还存在一些不够成熟的方面,但它的发展前景十分广阔,微生物菌群培养组学方法和其他研究方法的互补已经逐渐成为发展兽医微生物学新的突破口。     

肠道免疫屏障功能损伤的研究进展

肠黏膜免疫屏障作为肠屏障中的第一道防线,有效地保护肠黏膜的完整性,使其不受有害病原体的感染。但在肠黏膜缺血、缺氧、肠道细菌移位、肠内营养障碍、细胞凋亡等状态下,肠黏膜免疫屏障将出现一系列生理和病理变化,导致肠黏膜萎缩和通透性增大。为此,文章从几种损伤机制的角度,对造成肠道免疫屏障损伤的因素及其作用机理作一综述。     

Identification of Suspected Stem/progenitor Cells in Bovine Mammary Epithelial Cells Culture System

To develop the potential function of dairy cow mammary stem cells (DCMECs) in regulation of lactation,we identify putative DCMECs which were BrdU label retaining epithelial cells,at the same time,analysis the location of two new mammary stem cells molecular marks FNDC3B and PROCR to verify the feasibility of them to indicate DCMECs.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-1 and their receptors were detected along with cell passage by Real-time quantitative PCR.The results showed that the proportion of BrdU label-retaining epithelial cells was nearly 0.4% after 25 d continuous culture (passaged 4 times) and few cells were positive for FNDC3B or PROCR.Moreover,we observed the BrdU labelled epithelial cells by asymmetric division.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-Ⅰ and their receptors in primary and passage cells were extremely significant difference(P<0.01).DCMECs would rapidly lose some physiological characteristics and the ability of milk synthesis when not under the condition of induction of lactation differentiation,but a certain percentage of mammary stem/progenitor cells will be retained,whose potential effects on the regulation of lactation and mammary acinar remodeling were worthy of attention.     

C型利钠肽及其受体在水牛卵泡中的表达分析

为了探明C型利钠肽（C-type natriuretic peptide,CNP）及其受体（natriuretic peptide receptor,NPR）在水牛卵泡中的表达模式,本研究首先采用实时荧光定量PCR技术检测水牛卵巢中利钠肽家族主要成员A型、B型、C型利钠肽（ANP、BNP、CNP）及其Ⅰ型、Ⅱ型受体（NPR1、NPR2）的mRNA表达情况,然后利用免疫组化技术对水牛卵泡中CNP及NPR2蛋白进行定位,最后利用实时荧光定量PCR技术检测颗粒细胞和卵丘细胞中CNP及NPR2的mRNA表达规律。结果显示,水牛卵巢主要表达CNP及NPR2,且在各级卵泡中均有表达,其中,CNP主要在壁层颗粒细胞中表达,NPR2主要在卵丘细胞中表达;颗粒细胞上CNP mRNA表达水平显著高于卵母细胞周围的卵丘细胞（P<0.05）,而卵丘细胞上NPR2 mRNA表达水平显著高于颗粒细胞（P<0.05）。综上所述,在利钠肽主要家族成员和受体中,CNP和NPR2在水牛卵巢中呈现强表达,CNP主要在壁层颗粒细胞中表达,而NPR2主要在卵母细胞周围的卵丘细胞中表达。