呼伦贝尔市首例羊源荚膜D群多杀性巴氏杆菌的分离与鉴定

为了确定呼伦贝尔市两个羊场的羊只出现体温升高、精神沉郁、食欲废绝、咳嗽、腹泻甚至死亡病因,本研究以病羊病变组织为研究对象,采用常规细菌培养法和多杀性巴氏杆菌种特异性基因KMT1引物以及cap A、cap B、cap D、cap E、cap F荚膜血清型特异性基因引物进行双重PCR扩增,确定分离菌的荚膜血清型,同时应用纸片扩散法对分离菌进行药物敏感性试验。结果显示:从病变的肺组织中分离到1株菌落为灰白色、露珠状、不溶血,革兰氏染色阴性球状短杆菌;通过序列比对及遗传进化树分析,该分离株与印度分离株巴氏杆菌同源性高达100%;本研究只扩增出多杀性巴氏杆菌荚膜血清D群;分离菌株对青霉素、复方新诺明、克林霉素等耐药,对诺氟沙星、卡那霉素、环丙沙星等药物已不敏感。本研究结果表明,我们首次从呼伦贝尔市病羊体内分离到荚膜血清D群多杀性巴氏杆菌。     

1株羊源多杀性巴氏杆菌的分离鉴定及生物学特性研究

为了确定和分析新疆某羊场患呼吸道疾病病例的细菌性病原，采集病死羊的肺脏、胸腔积液等样品进行细菌分离，通过生化试验、16S rRNA基因测序、特异性引物PCR鉴定、药物敏感性试验、毒力基因测定、对小鼠半数致死量(LD₅₀)测定等方法对分离株进行生物学特性研究。结果显示，从病羊胸腔积液中分离鉴定出一株荚膜血清D型多杀性巴氏杆菌，将其命名为Pmh7。Pmh7对链霉素、头孢呋辛等10种抗菌药物敏感，对万古霉素、杆菌肽等5种抗菌药物耐药；携带6个毒力基因(fimA、nanB、ompH、sodC、Oma87、sodA),对小鼠的LD₅₀为2.3×10~6 CFU。结果表明，引发该羊场呼吸系统疾病的病原为荚膜D型多杀性巴氏杆菌，有较强的致病性。本研究结果可为防治羊巴氏杆菌病提供参考，为巴氏杆菌病的流行病学监测奠定基础。     

羊源D型多杀性巴氏杆菌的分离鉴定及毒力基因检测

为了确定石河子地区规模化羊场出现呼吸道症状死亡羊的细菌性病原,采用常规细菌分离鉴定方法、细菌16SrRNA序列分析以及多杀性巴氏杆菌特异性基因kmt,从病变肺组织分离鉴定细菌,利用5个荚膜血清型特异性基因确定其血清型,扩增分离株的16个毒力相关基因,分析分离菌致病性和对常用抗菌药物的耐药性。结果表明,从病羊的病变肺组织中分离鉴定到一株血清D型多杀性巴氏杆菌,具有较强的致病性;携带8个毒力相关基因,其片段序列与NCBI上己公布的多杀性巴氏杆菌参考株同源性在99%以上;分离株对青霉素、林可霉素、庆大霉素、复方新诺明耐药,对其他23种药物敏感。     

三株鸭源荚膜A型多杀性巴氏杆菌的分离与鉴定

从疑似禽霍乱死亡鸭分离到3株细菌,经PCR法鉴定为荚膜A型多杀性巴氏杆菌,纸片法进行药物敏感试验,发现3株细菌同时敏感的药物有恩诺沙星、多黏菌素B、复方新诺明和头孢唑啉,3株细菌同时耐受的药物有链霉素、四环素、克林霉素和青霉素。本研究结果可为禽源多杀性巴氏杆菌的血清流行病学和禽霍乱的临床治疗提供参考。     

一株兔源荚膜血清D型多杀性巴氏杆菌的分离和鉴定

为确定山东某兔场母兔子宫内膜炎病原,从病死兔子宫内膜炎的子宫脓液中分离到一株细菌,并对细菌进行了鉴定。结果显示,通过瑞氏染色、特异性PCR以及荚膜血清分型PCR鉴定,确定该株致病菌为荚膜血清D型的多杀性巴氏杆菌,命名为SDZG0324;通过多位点序列分析,确定该株致病菌为ST11型;利用Mega X生物学软件进行基因进化分析,表明该菌与多杀性巴氏杆菌经典菌株处于同一大分支,但是却为独立的分支;药敏试验表明,该菌对氨苄西林和庆大霉素2种药物耐药,对青霉素、环丙沙星和复方新诺明等6种抗生素敏感。说明在发生兔子宫内膜炎的样本中分离到荚膜D型多杀性巴氏杆菌,丰富了子宫内膜炎病原学资料,同时为该病的防控提供了理论指导。     

猪源多杀性巴氏杆菌的分离鉴定及其荚膜血清型鉴定

从病死母猪肺脏中分离到一株革兰氏阴性小杆菌,用生理生化鉴定、药敏试验、致病性试验和PCR鉴定方法对分离菌株进行鉴定,并用多杀性巴氏杆菌荚膜分型引物对分离株的荚膜血清型进行鉴定。结果表明:本菌为猪多杀性巴氏杆菌,对多种抗生素高度敏感,对小白鼠有强致病性;PCR扩增16SrDNA基因获得1415bp片段,分离株的16SrDNA核苷酸序列与多杀性巴氏杆菌(AY078999)的同源性为99%,因此该分离菌株被鉴定为致病性巴氏杆菌,命名为YN20110122株;本菌分离株为荚膜A型血清型多杀性巴氏杆菌。     

上海市猪源多杀性巴氏杆菌分离鉴定及其耐药性分析

本研究旨在调查上海市猪源多杀性巴氏杆菌对临床常用抗菌药的耐药情况,为该病的药物预防与治疗提供科学依据。2014~2015年在上海市11个规模化养猪场采集疑似多杀性巴氏杆菌感染的病料共计116份,采用细菌分离培养及PCR方法,分离鉴定出20株多杀性巴氏杆菌。采用纸片法测定了分离菌株对20种临床常用抗菌药的敏感性,结果表明,所有菌株对阿莫西林克拉维酸、头孢曲松、恩诺沙星、氟苯尼考、阿奇霉素和多黏菌素B敏感,这6种药物可作为本地区控制本病的首选药物。然而,超过50%的菌株对青霉素(65.0%)、复方新诺明(65.0%)、氨苄西林(60.0%)、链霉素(60.0%)、四环素(60.0%)耐药。耐药谱分析显示,85.0%的菌株对4~7种抗菌药耐药。本研究结果表明上海市猪源多杀性巴氏杆菌对临床常用抗菌药物耐药较严重,应加强养殖场猪多杀性巴氏杆菌的耐药性监测与防控。     

肉牛多杀性巴氏杆菌的分离鉴定及药敏分析

为调查多杀性巴氏杆菌在河北省部分地区肉牛群中的流行情况,本课题组从规模化肉牛养殖场中采集患有呼吸道综合征的病牛鼻腔拭子进行细菌分离鉴定、荚膜血清分型、毒力基因检测、动物攻毒试验及药物敏感性分析。结果显示,从105份鼻腔拭子中共分离了25株多杀性巴氏杆菌;菌株荚膜分型均为A型,携带ptfA、exbB和nanH等多种毒力基因,LD_(50)显示多数菌株毒力较强;分离菌株对大多数药物敏感性较高,但对复方新诺明、磺胺间甲氧嘧啶、磺胺二甲氧嘧啶和林可霉素药物耐药性较高,耐药率达到80%以上。     

猪多杀性巴氏杆菌的分离鉴定及oppA基因遗传进化分析

【目的】了解福建省猪场猪多杀性巴氏杆菌（Pasteurella multocida,Pm）的流行及oppA基因遗传进化情况。【方法】本研究采用细菌分离培养、生化试验、16S rRNA PCR扩增测序、PCR荚膜分型、oppA基因克隆及相似性分析、动物回归试验等方法对分离菌株进行鉴定和分析。【结果】本研究共分离到10株菌,分离菌在血平板上形成淡灰白色、湿润光滑、奶油露珠状菌落;分离菌株能酵解蔗糖、果糖、麦芽糖和甘露醇,不能分解葡萄糖、枸橼酸盐、乳糖、硫化氢等,与多杀性巴氏杆菌生化特性基本一致;分离菌株16S rRNA序列与GenBank中登录的多杀性巴氏杆菌相似性达99.9%以上,10株分离菌均为多杀性巴氏杆菌;PCR荚膜分型显示,6株分离菌为荚膜A型,4株为荚膜D型;基于oppA基因的遗传进化树显示,10株分离菌均位于同一分支内;动物回归试验结果显示,在24 h内攻毒小鼠死亡率较高（21/30）,分离菌有较强的致病力。【结论】福建省猪场猪多杀性巴氏杆菌流行菌株的荚膜血清型主要是A和D型,且大部分菌株都来源于共同的祖先,本研究结果丰富了猪多杀性巴氏杆菌的流行病学资料,并为该病的防控奠定基础。     

产毒素多杀性巴氏杆菌的鉴定

采用菌落多重PCR方法对分离保存的28株多杀性巴氏杆菌进行种型和毒素基因的检测,结果表明,菌株C51-6、M-4和P-2237为产毒素多杀性巴氏杆菌,菌株C51-6和P-2237为荚膜血清D型,菌株M-4为荚膜血清A型。同时用金黄色葡萄球菌抑制试验、中性吖啶黄沉淀试验和豚鼠皮肤坏死试验对PCR方法进行了验证。基于对甘露醇、卫茅醇、山梨醇、海藻糖的发酵能力和产生鸟氨酸脱羧酶的特性,3株菌株鉴定为多杀性巴氏杆菌多杀亚种。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

中华人民共和国农业部公告 第267号

为贯彻落实《兽药生产质量管理规范》(简称《兽药GMP》),进一步推动兽药GMP实施进程,我部制定了《兽药生产质量管理规范检查验收办法》,现予公告。本公告自2003年6月1日起施行。附件:兽药生产质量管理规范检查验收办法二○○三年四月十日第一章 总则 第一条 为推动《兽药生产质量管理规范》(以下简称兽药GMP)的实施,规范兽药GMP检查验收工作,制定本办法。 第二条 农业部负责全国兽药GMP管理和检查验收工作;负责制修订兽药GMP检查验收管理规定;负责兽药GMP检查员队伍建设和监督管理工作,负责国际兽药贸易中GMP互认工作。 …     

鸡败血支原体垂直传播模式及其阻断方法

以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.