固相萃取-超高效液相色谱串联质谱法快速测定猪尿中的赛庚啶

建立了猪尿中赛庚啶的固相萃取-超高效液相色谱串联质谱的测定方法。猪尿样品经酸化,混合型阳离子交换固相柱萃取后,以超高效液相色谱串联质谱测定,外标法定量。本方法的测定线性范围为0.2～5.0μg/L,在0.2、0.4、2.0μg/L低、中、高三个浓度的回收率为80%～120%,批内、批间精密度小于20%。本方法简便、快速、灵敏,经实际样品分析,适用于猪尿中赛庚啶的定量测定和确证。     

高效液相色谱法检测动物脂肪组织中辛硫磷残留量的研究

建立了高效液相色谱法检测动物脂肪组织中辛硫磷残留量的方法.均质后的动物脂肪组织在微波液化后,经乙腈提取,正己烷脱脂,旋转蒸发浓缩后净化,残余物用80％乙腈-水溶解,供高效液相色谱定量(紫外检测器)检测.该方法在浓度0.05 ～2 μg/mL之间呈线性相关,相关系数(R2)为0.9999;在空白脂肪组织样品中添加20(定量限)、200、400、800 μg/kg的辛硫磷,回收率为80％～110％,相对标准偏差(RSD)小于10％.该方法简单快速,灵敏度高,重现性好,适合动物组织中辛硫磷残留量的测定.     

褪黑素的添加减轻猪精原干细胞的氧化损伤

旨在研究褪黑素(melatonin,MT)对体外培养猪精原干细胞(spermatogonial stem cells,SSCs)的作用机制.本研究采集3头7日龄健康大白公猪睾丸,利用差速贴壁法获得SSCs.后经形态学观察、碱性磷酸酶染色、标记基因检测及免疫荧光染色鉴定后以SSCs作为试验材料,设置MT浓度梯度(0、50...     

结肠小袋纤毛虫体外培养特性的初步观察

目的观察结肠小袋纤毛虫（Balantidium coli）在体外培养条件下的生长情况。方法取粪样分别置于4℃～8℃冰箱,28℃和37℃恒温培养箱中,观察记录虫体数量,统计不同温度条件下滋养体生存时间。28℃恒温条件下,将粪液加入不同稀释液（蒸馏水、生理盐水及洛克氏液）中,观察滋养体的活力及数量;采用同样的观察方法,统计滋养体在双相培养基以及免血水培养基内的生存时间。在28℃和37℃恒温条件下,分别观察滋乔体在RSS培养液（Ringer液＋血清＋米粉）及不加米粉的RSS培养液内的生存时间。结果结肠小袋纤毛虫滋养体在体外的最适生存温度为28℃,在洛克氏夜中生存时间明显长于生理盐水和蒸馏水,在RSS培养基及双相培养基中可在培养24h时达到高峰。在含有诺氟沙星的兔血水培养基中,滋养体的数量可在培养后96h达到高峰,生存时间可达到180h。结论结肠小袋纤毛虫体外培养受很多因素的影响,有待于进一步研究。     

用地高辛标记的核酸探针检测猪伪狂犬病毒野毒感染的研究

利用PCR技术从带有伪狂犬病毒（PRV）gE基因的重组质粒pMD18-T-gE中扩增回收约304bp大小的片段,并制备出地高辛标记的gE基因核酸探针。特异性检测结果表明,该探针能与重组质粒DNA发生特异性杂交,而与对照的PRVBartha-k61株疫苗毒DNA、猪细小病毒（PPV）DNA、猪圆环病毒（PCV）DNA、猪繁殖与呼吸综合征病毒（PRRsV）cDNA、猪瘟病毒（CSFV）cDNA的杂交反应均为阴性;敏感性检测结果表明,该探针对PRV野毒的最低检出量为4pg。应用该探针对11份繁殖障碍病料进行了杂交检测,共检出4份阳性病料,该结果与PCR检测结果一致,表明该核酸探针可用于猪伪狂犬病野毒感染的临床诊断。     

Genetic and pathobiologic characterization of H3N2 canine influenza viruses isolated in the Jiangsu Province of China in 2009-2010

The newly emerging canine influenza virus (CIV) causes considerable concerns for both veterinary and public health. During 2009-2010, six strains of H3N2 influenza virus were isolated from dogs in Jiangsu Province, China. Sequence and phylogenetic analysis of eight gene segments revealed that the six viruses were most similar to a recent canine-derived subtype H3N2 influenza virus isolated in cats from South Korea, which originated from avian strain. By comparing the deduced amino acid sequences of the hemagglutinin 1 (HA1) and neuraminidase (NA) genes of the six Jiangsu isolates against the most similar avian strains, we found that all isolates had several common mutations at the receptor-binding sites, potential glycosylation sites and cleavage site in HA1, and antigenic sites in both the HA1 and NA segments. Significantly, a unique two amino acid insertion in the NA stalk was found. Experimental infection of BALB/c mice revealed that viral RNA could be detected in the major rodent organs, such as brain, heart, spleen, kidney, liver and intestine, as well as the lung. All the sampled organs from infected mice showed significant lesions and viral antigen staining. This study highlights the potential of domesticated animals to become a reservoir for influenza virus and the need for surveillance programs to detect cross-species transmission.     

绵羊痘病毒山东流行株的分离鉴定与T4肽基因序列分析

从山东东营疑似绵羊痘病羊的肺脏组织中分离到1株病毒。取疑似绵羊痘病羊组织病料研磨、冻融、离心后分别接种11日龄SPF鸡胚绒毛尿囊膜和牛睾丸继代细胞,鸡胚接种部位出现明显的痘斑;牛睾丸细胞出现聚集、变圆等明显的细胞病变。利用1对绵羊痘病毒T4肽基因特异性引物进行了PCR扩增,获得与预期大小一致的约300 bp的片段。将所得序列与GenBank收录的绵羊痘病毒、山羊痘病毒等毒株相关序列进行比较分析。结果表明,该分离株与国外其他绵羊痘病毒株具有较高的同源性,达97.4%～99.3%;与山羊痘病毒株同源性为96.4%。通过试验初步证明所分离的病毒为绵羊痘病毒,并将该毒株命名为绵羊痘病毒DY株。     

草鸡源H9亚型禽流感病毒的分离与鉴定

2010年4月,安徽省某草鸡养殖场发生一起以严重呼吸困难、急性死亡为特征的病例。取发病典型的鸡内脏,按常规方法处理后,接种10日龄SPF鸡胚,经有限稀释法纯化,获得一株病毒,通过HA、HI、分子生物学以及动物回归等试验,最终确诊该病为H9亚型禽流感。     

马立克强毒MDV-GA对BWEL-SPF鸡的感染试验

134只1日龄BWEL-SPF鸡随机分成2组,攻毒组84只腹腔注射2 000 PFU的MDV-GA细胞毒0.2 mL,对照组50只注射同等剂量的病毒稀释液。结果显示,攻毒后27 d攻毒组鸡只开始发病并持续到攻毒后51 d,发病高峰期在攻毒后28~42 d。经过60 d的观察发现,该毒株可以引起很高的肿瘤发生率(61.90%),死亡率为75.00%。肿瘤主要出现在肾脏、心脏、肝脏、性腺、脾脏、肺、胸腺、皮肤、腺胃等组织,其中以肾脏、心脏、肝脏、性腺、脾脏肿瘤发生率较高,分别为:36.90%,28.75%,25.00%,21.43%,17.86%。其中2个以上组织发生肿瘤的个体占44.08%;3个以上组织发生肿瘤的个体占27.38%。多数肿瘤个体同时伴有胸腺、法氏囊萎缩,脾脏、性腺肿大等症状。经过对发病鸡的肾脏作组织切片观察,发现其基本结构已经消失,伴随大量的淋巴细胞和浆细胞浸润,呈现典型的马立克症状。结果表明,利用2 000 PFU的MDV-GA毒株感染BWEL-SPF鸡可以得到大量的肿瘤样本,可用于探讨马立克淋巴瘤内部的基因变化。     

Antegrade pyelography for suspected ureteral obstruction in cats: 11 cases (1995-2001)

OBJECTIVE: o determine sensitivity and specificity of radiography, ultrasonography, and antegrade pyelography for detection of ureteral obstructions in cats. DESIGN: Retrospective study. ANIMALS: 11 cats. PROCEDURE: Medical records of cats that had radiography, ultrasonography, and antegrade pyelography performed for suspected ureteral obstructions were examined. Ultrasound-guided pyelocentesis and fluoroscopic-assisted antegrade pyelography were performed on 18 kidneys in 11 cats. Obstructive ureteral lesions were confirmed in all cats by surgical or necropsy examination. Sensitivity and specificity of survey radiography, ultrasonography, and antegrade pyelography for identification of ureteral obstructions were calculated. Surgical or necropsy findings were used as the standard for comparison. RESULTS: All cats were azotemic. Mean +/- SD serum creatinine and BUN concentrations were 10.2 +/- 6.1 and 149 +/- 82 mg/dL, respectively. Fifteen of 18 ureters were found to be obstructed at surgery or necropsy. Sensitivity and specificity were 60 and 100% for radiography and 100 and 33% for ultrasonography, respectively, in identification of ureteral obstructions. Leakage of contrast material developed in 8 of 18 kidneys during antegrade pyelography and prevented diagnostic interpretation in 5 of 18 studies. For the 13 diagnostic studies, specificity and sensitivity were 100% by use of the antegrade pyelography technique. Correct identification of the anatomic location of the ureteral obstruction was obtained in 100% of diagnostic antegrade pyelography studies and in 60% of radiography or ultrasonography studies. CONCLUSIONS AND CLINICAL RELEVANCE: Antegrade pyelography can be a useful alternative in the diagnosis and localization of ureteral obstructions in azotemic cats, although leakage of contrast material may prevent interpretation of the study.