西宁市售鲤鱼鳃中志贺氏菌的检测及血清学分型

志贺氏菌是最常见的肠杆菌科的病原菌,可引起人类肠道传染病。应用常规细菌分离法对西宁市售的80份鲤鱼鳃样品进行了志贺氏菌的检测,同时对阳性菌株进行了血清学分型。结果显示,在80份鲤鱼鳃样品中共检出志贺氏阳性6份,阳性率为7.5%(6/80)。通过形态学观察、生化试验和血清学鉴定为志贺氏菌A群2株,其血清型为痢疾志贺氏菌1型;B群4株,其血清型为福氏志贺氏菌1b型。此次检测为以后西宁地区志贺氏菌流行病学的调查和相关疾病的防治提供了理论依据。     

福氏志贺菌2型多重PCR检测方法的建立

为构建一种快速、高效、特异性强的检测与鉴定福氏志贺菌的多重PCR方法,本试验以不同血清型志贺菌中的特有基因ipaH、gtrⅡ、gtrⅩ和R002为扩增目标来设计相对应的引物,在同一PCR体系中对福氏志贺菌进行检测。通过一系列探索及对反应条件的优化调整,最终建立了可在多种细菌中快速检测出福氏志贺菌2型（2a和2b）的方法。结果表明,应用多重PCR可快速、高效、准确的对福氏志贺菌进行验证和鉴别,对于预防实验室菌种污染和志贺氏菌病的临床诊断与治疗有着重要意义。     

牦牛源志贺菌分离鉴定、毒力基因检测与分型

本试验旨在阐明牦牛源志贺菌致病性及分子流行特性,为探索志贺菌流行途径,制定合理的防控策略提供新思路。2017年在甘肃、青海、西藏三省(区)共采集牦牛肛门棉拭子样品1 396份,通过选择培养基筛选、生化鉴定、血清凝集试验对分离菌株进行系统鉴定,应用PCR方法检测分离株中ipaH、ipaBCD、ial、sen、set1A、set1B和stx七种毒力基因流行情况。参照McMLST网站数据库提供的15对管家基因序列进行MLST分型;并参考美国CDC的PulseNet实验方法,用限制性内切酶NotⅠ和XbaⅠ分别对福氏志贺菌和宋内志贺菌染色体进行酶切,对这些分离菌株进行PFGE分析。结果显示,41株分离株符合志贺菌生化特征,分为4个生化表型,B3(36.59%)和B4(32.35%)为主要生化表型。血清凝集试验鉴定23株为福氏志贺菌,包括四个血清型1a(n=2)、2a(n=16)、2b(n=3)、Xv(n=2);18株为宋内志贺菌,分为Ⅰ相(n=12)和Ⅱ相(n=6)。共检测到6种毒力基因ipaH、ipaBCD、ial、sen、set1A、set1B,携带率分别为100%、92.68%、73.17%、70.73%、26.83%、26.83%。具有7种毒力基因型,其中VT5和VT7型为主要流行型,分别占43.9%和24.39%,同时携带两种及以上毒力基因的志贺菌占92.68%。41株志贺菌共分为10个ST型,其中ST100、ST116、ST155型为主要流行型。NotⅠ酶切的福氏志贺菌分为13个PT型,而XbaⅠ酶切的宋内志贺菌分为14个PT型。综上所述,牦牛源志贺菌生化表型、血清型、ST型和PT既存在多态性,又有优势流行型。本试验分离的志贺菌与人源志贺菌携带相同的毒力基因,其中ipaH、ipaBCD、ial、sen基因携带率较高,对公共安全具有潜在的威胁。     

牦牛源志贺菌分离鉴定、毒力基因检测与分型

本试验旨在阐明牦牛源志贺菌致病性及分子流行特性,为探索志贺菌流行途径,制定合理的防控策略提供新思路。2017年在甘肃、青海、西藏三省(区)共采集牦牛肛门棉拭子样品1 396份,通过选择培养基筛选、生化鉴定、血清凝集试验对分离菌株进行系统鉴定,应用PCR方法检测分离株中ipaH、ipaBCD、ial、sen、set1A、set1B和stx七种毒力基因流行情况。参照McMLST网站数据库提供的15对管家基因序列进行MLST分型;并参考美国CDC的PulseNet实验方法,用限制性内切酶NotⅠ和XbaⅠ分别对福氏志贺菌和宋内志贺菌染色体进行酶切,对这些分离菌株进行PFGE分析。结果显示,41株分离株符合志贺菌生化特征,分为4个生化表型,B3(36.59%)和B4(32.35%)为主要生化表型。血清凝集试验鉴定23株为福氏志贺菌,包括四个血清型1a(n=2)、2a(n=16)、2b(n=3)、Xv(n=2);18株为宋内志贺菌,分为Ⅰ相(n=12)和Ⅱ相(n=6)。共检测到6种毒力基因ipaH、ipaBCD、ial、sen、set1A、set1B,携带率分别为100%、92.68%、73.17%、70.73%、26.83%、26.83%。具有7种毒力基因型,其中VT5和VT7型为主要流行型,分别占43.9%和24.39%,同时携带两种及以上毒力基因的志贺菌占92.68%。41株志贺菌共分为10个ST型,其中ST100、ST116、ST155型为主要流行型。NotⅠ酶切的福氏志贺菌分为13个PT型,而XbaⅠ酶切的宋内志贺菌分为14个PT型。综上所述,牦牛源志贺菌生化表型、血清型、ST型和PT既存在多态性,又有优势流行型。本试验分离的志贺菌与人源志贺菌携带相同的毒力基因,其中ipaH、ipaBCD、ial、sen基因携带率较高,对公共安全具有潜在的威胁。     

野生草食动物源多杀性巴氏杆菌荚膜分型的研究

采用Carter荚膜群鉴定法对分离于我国黑鹿、白唇鹿、斑马和赤大袋鼠的56株多杀巴氏杆菌进行荚膜血清型鉴定,结果表明：B型占78．6％（44／56）,h型占14．3％（8／56）,D型占3．6％（2／56）,另有2株未能确定型,占3．6％（2／56）。其中荚膜B型是斑马、黑鹿和白唇鹿源多杀性巴氏杆菌的主要血清群,荚膜A型是赤大袋鼠源多杀性巴氏杆菌的主要血清群。黑鹿和白唇鹿源多杀性巴氏杆菌均存在2个血清群。     

一株鸡源志贺氏菌的分离、鉴定及耐药性分析

为了对鸡源志贺氏菌性腹泻进行快速准确的诊断和耐药性研究,试验对由贵州省动物疫病研究室提供的一株鸡源志贺氏菌进行进一步的分离、PCR鉴定及药敏试验。结果显示,经细菌分离培养、革兰氏染色及生化鉴定试验,该菌符合志贺氏菌的特点及其生化特性;PCR检测结果显示,针对志贺氏菌ipa H基因设计的一对引物可扩增出393 bp的目的条带,与预期大小一致,具有高度的特异性;对其耐药性的研究显示,该株志贺氏菌对参试的20种药物产生不同程度的耐药性。研究鸡源志贺氏菌的分离与鉴定中所结合的PCR方法具有快速、简单、特异性强的特点,能够迅速确定病原菌,并在药敏试验中发现了针对该菌敏感性较高、疗效更好的药物。结果为鸡源志贺氏菌耐药性的有关研究与临床用药提供了科学的参考依据。     

水貂志贺氏菌分离鉴定及药物敏感性分析

试验旨在对吉林省某水貂养殖场送检的3只具有典型腹泻症状的病死水貂的小肠和肠内容物样品进行细菌分离鉴定及耐药情况分析,为临床治疗提供参考。通过细菌分离纯化和PCR方法对分离菌株进行鉴定,对BALB/c小鼠进行菌液注射来检测菌株的致病性。采用K-B药敏法检测菌株对常用药物的敏感性,并通过PCR方法检测其耐药基因和Ⅰ类整合子的携带情况。结果显示,分离得到3株志贺氏菌,致病性检测试验显示分离菌可引起小鼠腹泻。药物敏感性试验结果显示,3株志贺氏菌对氟喹诺酮类药物、头孢类药物、氟苯尼考和多黏菌素较敏感,对氨基糖苷类、四环素、氯霉素、青霉素、氨苄西林耐药。耐药基因检测结果显示,3株志贺氏菌共检测出7种耐药基因bla_TEM-1、aadA1、aac（3'）-Ⅱc、aac（6'）-Ⅰb、aph（3'）-Ⅶ、tet（M）、cat2及携带aadA1基因盒的Ⅰ类整合子。结果表明,分离的3株志贺氏菌均具有致病性,可引起小鼠腹泻,主要表现为多重耐药现象,携带的耐药基因呈多样性,为临床治疗带来巨大影响。     

饲料中志贺氏菌的PCR快速检测方法的建立

为建立饲料中快速检测志贺氏菌的有效方法,本研究根据志贺氏菌(Shigella)侵袭性质粒抗原H基因(invasion plasmid antigen H,ipaH)设计了一对引物,PCR扩增片段长度为386bp。通过对引物的特异性检测发现,4株志贺氏菌的PCR结果均为阳性,12株非志贺氏菌的检测结果均未扩增出特异性片段。PCR对志贺氏菌纯培养物的检测灵敏度达120cfu/ml,检测快速、便捷。该方法操作简单、检验周期短、特异性和灵敏度高,能够快速地实现对饲料中志贺氏菌的检测和监控。     

细胞信号转导和骨架在人源福氏志贺菌侵袭鸡肠上皮原代细胞中的作用

为研究细胞信号转导和骨架在人源福氏志贺菌侵袭鸡肠上皮原代细胞中的作用,分别用酪氨酸蛋白激酶、酪氨酸蛋白磷酸酶、蛋白激酶C、Ca2+通道等信号转导抑制剂染料木黄酮、原钒酸钠、星状孢子素、硝苯地平和微丝蛋白聚合抑制剂细胞松弛素B处理鸡肠上皮原代细胞后,采用细胞免疫组织化学染色检测人源福氏志贺菌ZD02株对鸡肠上皮原代细胞侵袭率,用间接免疫荧光双重标记检测ZD02株侵袭鸡肠上皮原代细胞后F-肌动蛋白分布的变化。结果显示,染料木黄酮、硝苯地平和细胞松弛素B能显著抑制人源福氏志贺菌ZD02株内化,星状孢子素和原钒酸钠对ZD02株的内化无影响,人源福氏志贺菌侵袭鸡肠上皮原代细胞时F-肌动蛋白发生聚集。本研究表明人源福氏志贺菌ZD02株侵袭鸡肠上皮原代细胞过程中需要酪氨酸蛋白激酶、Ca2+通道活化以及细胞微丝重排。     

志贺氏菌属环介导恒温扩增快速检测方法的建立及初步应用

为建立快速检测志贺氏菌属的方法,本研究针对其ipaH基因,建立了志贺氏菌属环介导恒温护增(LAMP)快速检测方法。利用该方法对50株细菌进行检测,结果显示7株志贺氏菌均为阳性,其它菌均为阴性,表明该方法具有高度特异性。灵敏度试验显示,对志贺氏菌纯培养菌的检测灵敏度为28cfu/mL,对污染食品中志贺氏菌的检测灵敏度为35cfu/mL。利用该方法对195份涉及肉类、奶类、豆制品及人工污染样品进行检测,共检出29份阳性样品,与细菌分离鉴定检测结果一致。该LAMP方法操作简便、特异性强、灵敏度高且成本低廉,具有良好的应用前景。     

Comparison of in vitro and in situ methods in evaluation of forage digestibility in ruminants

The objective of this study was to compare the application of different in vitro and in situ methods in empirical and mechanistic predictions of in vivo OM digestibility (OMD) and their associations to near-infrared reflectance spectroscopy spectra for a variety of forages. Apparent in vivo OMD of silages made from alfalfa (n = 2), corn (n = 9), corn stover (n = 2), grass (n = 11), whole crops of wheat and barley (n = 8) and red clover (n = 7), and fresh alfalfa (n = 1), grass hays (n = 5), and wheat straws (n = 5) had previously been determined in sheep. Concentrations of indigestible NDF (iNDF) in all forage samples were determined by a 288-h ruminal in situ incubation. Gas production of isolated forage NDF was measured by in vitro incubations for 72 h. In vitro pepsin-cellulase OM solubility (OMS) of the forages was determined by a 2-step gravimetric digestion method. Samples were also subjected to a 2-step determination of in vitro OMD based on buffered rumen fluid and pepsin. Further, rumen fluid digestible OM was determined from a single 96-h incubation at 38°C. Digestibility of OM from the in situ and the in vitro incubations was calculated according to published empirical equations, which were either forage specific or general (1 equation for all forages) within method. Indigestible NDF was also used in a mechanistic model to predict OMD. Predictions of OMD were evaluated by residual analysis using the GLM procedure in SAS. In vitro OMS in a general prediction equation of OMD did not display a significant forage-type effect on the residuals (observed - predicted OMD; P = 0.10). Predictions of OMD within forage types were consistent between iNDF and the 2-step in vitro method based on rumen fluid. Root mean square error of OMD was least (0.032) when the prediction was based on a general forage equation of OMS. However, regenerating a simple regression for iNDF by omitting alfalfa and wheat straw reduced the root mean square error of OMD to 0.025. Indigestible NDF in a general forage equation predicted OMD without any bias (P ≥ 0.16), and root mean square error of prediction was smallest among all methods when alfalfa and wheat straw samples were excluded. Our study suggests that compared with the in vitro laboratory methods, iNDF used in forage-specific equations will improve overall predictions of forage in vivo OMD. The in vitro and in situ methods performed equally well in calibrations of iNDF or OMD by near-infrared reflectance spectroscopy.     

大庆市羊东毕吸虫病流行病学调查

对大庆市羊东毕吸虫病的流行病学进行了调查研究。结果表明，大庆有3种东毕吸虫，即程氏东毕吸虫（O.cheni)、土耳其斯坦东毕吸虫（O.turkestanica)和土耳斯坦东毕吸虫结节变种（O.turkestanican var.thuberculata)；中间宿主螺类有1种，即卵萝卜螺；东毕吸虫病在大庆分布很广，所调查的4区2县均有发生，羊东毕吸虫平均感染率分别为50．33％，平均死亡率为21．12％，大庆羊急性病例发病时间为8月上旬至10月下旬，慢性病例为11月上旬至翌年2月中旬。     

Use of cyclocryotherapy in management of glaucoma in dogs

In 5 cases of glaucoma (2 from trauma, 2 from narrowed drainage angles, 1 secondary to lens extraction), cyclocryotherapy was used to control intraocular pressure. In all cases the intraocular pressure decreased, with the usual result being a cosmetic and painless but blind eye.     

中国圈养野生动物疫苗使用调查

2005年对中国动物园协会单位中的18家动物园饲养野生动物疫苗免疫情况进行调查。受调查动物园在野生动物疫苗使用和动物种类方面具有一定的代表性。调查结果显示动物园动物现共使用18类36种疫苗,预防31种疫病。其中哺乳动物使用14类24种疫苗,预防24种疫病;禽类使用4类12种疫苗,预防7种疫病。使用范围最广的有禽流感、犬瘟热疫苗等、新城疫疫苗、猫瘟热疫苗等。共有24目58科动物接种疫苗,其中食肉动物类1目8科,食草动物类3目9科,杂食动物类2目3科,禽类18目38科。研究结果提示应加强动物园之间疫病信息交流、防疫资源的利用、疫病监测和研究,加大对动物园动物疫病的研究投入,逐步建立动物园动物统一的防疫规程。     

Estimate of economic impact of fowl cholera in turkeys in Georgia in 1986

Information gathered from cases of fowl cholera (FC) in commercial turkey flocks through case records, flock records, and telephone and mail surveys was used to estimate disease costs. The cost to the Georgia commercial turkey industry in 1986 from preventive measures, treatment of outbreaks, and production losses from the disease was estimated at $634,545. The cost of FC per kg of live production was estimated to be $0.015.     

ELISA试验在防制猪伪狂犬病中的应用

猪伪狂犬病是由伪狂病病毒(Pseudorabies virus PRV)引起的急性传染病。其特征是成年猪常为隐性感染，可有流产、死胎、呼吸系统症状，新生仔猪除神经症状外还可侵害消化系统，并伴大批死亡。本病已经给全世界的养猪业造成严重的损失，我国许多地区已报道该病。为进一步探讨该病的控制、净化措施，我们对12个养猪场共856分血清采用美国嗍公司的ELISA试剂盒进行了抗体检测。     

Occurrence of MRSA in pigs and in humans involved in pig production--preliminary results of a study in the northwest of Germany

In 2007, 678 pigs of all age groups out of 347 different farms from Lower Saxony and Northrhine-Westphalia and 86 persons occupationally exposed to pigs were investigated for their nasal colonisation with methicillin-resistant Staphylococcus areus (MRSA) by the Field Station for Epidemiology of the University of Veterinary Medicine Hannover and the Robert Koch-Institute. At the individual animal level, a frequency of positive results of 13% (n = 85 positive animals) and at the herd level, a frequency of positive results of 18% (n = 62 positive herds) were found. All isolates were assigned to the Multilocus Sequence Typing Type ST398. Within MRSA-positive herds, there were more MRSA-negative than MRSA-positive animals. Among the occupationally exposed persons (veterinarians, laboratory personnel and meat inspection personnel), 20 persons (23%) showed a nasal colonisation with MRSA ST398. A quite strong association between the intensity of the contact to pigs with the frequency of nasal colonisation in the occupationally exposed persons was detected. None of the animals or the humans nasally colonised by MRSA ST398 showed any clinical symptoms of a staphylococcal infection. Conclusions are drawn on the herd and intra-herd prevalence of the nasal colonisation of pigs with MRSA ST398 in pigs, but especially on which questions need to be addressed by further research.