短乳杆菌的分离鉴定及其对鸡肠粘膜SlgA分泌的影响

用MRS培养基,从SPF鸡的消化道分离到1株乳酸杆菌.该菌生长速度快,耐酸,耐胆汁,对肠道粘膜有较强的粘附力,能提高鸡肠粘膜的SIgA滴度.对该菌的形态特征、培养特性、生理生化特性、药物的敏感性及G+C等进行试验,证明该菌为短乳杆菌.该菌可作为禽用益生素的候选菌株.     

鸡源大肠埃希氏菌某些生物学特性

对鸡源大肠埃希氏菌的某些生物学特性进行了研究,包括对１日龄鸡的致病性、溶血性、菌毛的表达、血凝性、对鸡胚成纤维细胞（ＣＥＦ）的粘附特性、体内外与鸡气管粘膜的粘附特征、质粒特征、全菌蛋白ＳＤＳ－ＰＡＧＥ电泳及免疫转印等。结果表明,鸡源大肠埃希氏菌的溶血性、质粒特征、对ＣＥＦ的粘附特性与其致病性无关;菌毛与致病性有关,大部分菌株表达Ⅰ型菌毛;血凝能被Ｄ－甘露糖抑制,不同菌株血凝谱具有差异;体内外与气管粘膜的粘附特性与菌株致病性有关;在全菌蛋白ＳＤＳ－ＰＡＧＥ电泳中,相对分子质量约为４０７００的多肽仅存在于具有致病性的菌株中,经免疫转印证实,该多肽为致病性大肠埃希氏菌所特有     

艾维茵肉仔鸡爆发盲肠肝炎的诊治

葡萄球菌广泛存在于自然界中，同时也是动物体表及粘膜的常带菌，该菌主要通过溃烂的皮肤和粘膜侵人鸡的体内而引起发病。     

鸡源肺炎克雷伯氏菌的致病性和生物学特性研究

本文系统地报告鸡源肺炎克雷伯氏菌通过口腔接种，种蛋及种蛋蛋壳用新洁而灭消毒而后涂沫该菌进行的感染试验，试验表明该菌不产生血凝素和溶血素，但具有肠毒素和致细胞病变因子等生物学特性。     

体内繁殖禽巴氏杆菌的提取及其交叉保护特性

通过不同攻毒途径、不同攻毒剂量、不同采血时间及不同采血途径的试验表明，以翅静脉途径攻击多杀巴氏杆菌，剂量10亿／只，在鸡濒死挣扎时的即刻颈静脉无菌放血，可以自鸡体采集到最大量的含菌血，每只鸡37～39mL，再将血液进行差速和蔗糖密度梯度离心，可最大限度地提取到血液中的巴氏杆菌，每只鸡300～400亿的总菌量。该结果与报道的提取方法相比，效率提高了1000倍以上。提取的细菌灭活后免疫鸡，经异型菌交叉攻毒试验表明，体内繁殖的巴氏杆菌具有交叉保护特性，而体外培养菌则没有。用该方法提取的巴氏杆菌，其总蛋白和总糖含量均高于培养基培养菌，提示二者之间的差异可能与交叉保护特性存在着一定的相关性。     

1例鸡白痢沙门氏菌的分离鉴定与耐药性分析

为查明河南省新乡市某规模化蛋鸡场疑似沙门氏菌感染病例的病原，进而制定合理的治疗方案，本研究无菌采集疑似病雏鸡肠管样品，用革兰氏染色、培养特性观察、生化试验、PCR方法对其进行确诊，并进行病原回归试验，采用药敏纸片琼脂扩散法（K-B法）分析分离菌对19种常见抗生素的耐药性。结果显示，本试验成功分离了一株革兰氏阴性短杆菌，该分离菌符合鸡白痢沙门氏菌的培养特性和生化特性；PCR成功扩出invA基因，通过与GenBank数据库比对分析确定该细菌为鸡白痢沙门氏菌；用分离细菌株感染SPF鸡能复制出与自然感染一致的病例，说明鸡白痢沙门氏菌是造成本养鸡场雏鸡发病的主要病原；分离株具有较强的致病性和多重耐药性，对阿米卡星、苯唑青霉素、青霉素耐药，对复达欣、氨苄青霉素、头孢曲松等药物敏感，将敏感抗生素用于临床治疗效果明显。结合以上结果，本研究提出该病的具体防治措施，并取得了较好的效果，为鸡白痢沙门氏菌的分离鉴定及防治提供了参考，对鸡白痢沙门氏菌病病原的早期诊断和治疗具有重要临床意义。     

鹑梭状芽胞杆菌的分离鉴定

用色氨酸磷酸琼脂培养基，采用厌氧培养的方法，对从泰安、菏泽、聊城等地送检的疑似鸡溃生肠炎的病死鸡进行了鹑梭状芽胞杆菌的分离，并对谝菌的培养特性、生化特性、对药物的敏感性、致病性等进行了测定。     

口服腔上囊提取液对鸡生长发育及抗大肠埃希氏菌感染的影响

对１日龄ＳＰＦ鸡通过口服腔上囊提取液，４ｄ后剖杀称重及对腔上囊进行组织学检查。结果：与对照组相比，试验组鸡增重较快，腔上囊重无明显差别；组织学检查试验组鸡腔上囊发育较快，腔上囊淋巴滤泡增多、滤泡间隙变窄、滤泡皮髓质分界明显，其内充满大量淋巴细胞和网状内皮细胞。人工感染大肠埃希氏菌试验显示，腔上囊提取液能推迟感染鸡的死亡时间。结果显示，通过粘膜接种途径不会影响该物质的生物学功能。     

牦牛肠毒素型大肠杆菌的分离鉴定和某些生物学特性的研究

从西藏地区腹泻死亡牦牛中分离出一株肠毒素型大肠杆菌并对其某些生物学特性进行了研究。结果表明，该菌在形态特`征培养特性和生化特性方面与大肠杆菌基本一致。血清学试验表明，该菌株O抗原属O148，K88、K99、987P单因子血清均不能凝集本菌；该菌不产生溶血素；对绵羊、豚鼠、马、鸡的红细胞表现强凝集，而K88、K99、987P抗血清均不能抑制其它对绵羊、豚鼠、马、鸡红细胞的凝集；该菌株在营养肉汤中经37℃，48小时培养表达菌毛；肌肉接种兔、腹腔接种小鼠均具有高致病性；乳鼠胃内投报试验和兔回肠结扎试验证明，该菌能产生热稳定肠毒素和热敏性肠毒素；分离菌株对恩诺沙星、环丙沙星、阿莫西林、羧苄青霉素等高度敏感，而对链霉素、四环素、土霉素等表现耐药性。     

葡萄球菌引发鸡腿病的原因及统治

葡萄球菌广泛存在于自然界中，同时也是动物体表及粘膜的常带菌，该菌主要通过溃烂的皮肤和粘膜侵入鸡的体内而引起发病。葡萄球菌使鸡最常受害的关节是跗关节、跖关节和趾关节。感染鸡早期跛行，不愿走动，或只用一条腿一瘸一拐地行走，严重时病鸡表现精神抑郁，受害关节肿胀发热，压迫可见痛感，病鸡很难吃料和饮水，最终死亡。切开病变关节可见关节腔内及周围组织中存有奶油状渗出物，病程长的鸡渗出物干稠，关节腔内有干酪样物质，一般可见跖骨后趾屈肌腱和跗关节上方的跖伸肌腱受损，轻者组织水肿充血，重者组织中有渗出物。由葡萄球菌…     

Enterotoxin activity of a Salmonella typhimurium of equine origin in vivo in rabbits and the effect of Salmonella culture lysates and cholera toxin on equine colonic mucosa in vitro

To determine whether a strain of Salmonella typhimurium (UCD 1755) of equine origin had enterotoxin activity, 2 ml of a cell-free culture lysate of strain UCD 1755 and approximately 10(9) viable strain UCD 1755 organisms were inoculated into ligated small intestinal segments of rabbits. Intestinal segments inoculated with viable strain UCD 1755 organisms and those inoculated with a cell-free culture lysate of strain UCD 1755 had significant (P less than 0.05) accumulations of fluid 10 hours after inoculation when compared with ligated intestinal segments either inoculated with sterile brain-heart infusion broth or left empty. There was not a statistically significant difference between fluid accumulation of intestinal segments inoculated with viable strain UCD 1755 and that of segments inoculated with a cell-free culture lysate of strain UCD 1755. The responses of equine colonic mucosa to culture filtrates of 2 strains of salmonella typhimurium (UCD 1755 and SL 1027) and purified cholera toxin were studied in vitro. Isolated smaples of colonic mucosa were incubated for 4 hours at 37 C in Krebs-Henseleit bicarbonate buffer (KHB) alone, KHB plus culture lysate of strain UCD 1755, KHB plus culture lysate of strain SL 1027, and KHB plus 1 microgram of cholera toxin/ml. Cyclic adenosine monophosphate (cAMP) content of each sample was determined.(ABSTRACT TRUNCATED AT 250 WORDS)     

仔猪小肠上皮细胞的体外培养及猪流行性腹泻病毒对其感染性研究

本研究旨在建立一种操作简单的仔猪小肠上皮细胞体外培养体系，为猪流行性腹泻病毒（porcine epidemic diarrhea virus，PEDV）的相关研究提供材料。研究以未吃初乳的新生仔猪作为肠道供体，采用肠腔面刮取肠黏膜和机械分离分散的方式进行原代细胞的体外分离。采用0.1%胰蛋白酶差速消化法进行仔猪小肠上皮细胞的纯化；比较新生弱仔猪和正常仔猪作为供体对原代小肠上皮细胞活性的影响；MTT法比较不同代次原代细胞的增殖活性；免疫荧光和实时荧光定量RT-PCR方法检测PEDV毒株CV777在原代小肠上皮细胞感染和增殖情况。结果显示，本研究建立的体外分离培养方法能获得增殖活性良好的原代小肠上皮细胞，具有明显的"S"型细胞增殖曲线。通过胰酶差速消化可得到纯度高、形态单一的小肠上皮细胞，同时细胞连续传代5次仍保持良好的增殖活性。弱仔猪和正常仔猪分离培养的小肠上皮细胞的增殖活性比较显示，两者并没有明显区别，这为降低原代细胞培养的成本提供新的思路。免疫荧光和实时荧光定量RT-PCR结果显示，PEDV可感染本方法分离培养的仔猪原代小肠上皮细胞，并在其中进行复制增殖。本研究建立了一种操作简单、实用性强、成本较低的仔猪原代小肠上皮细胞体外培养方法，该方法培养的原代细胞可作为PEDV分离培养和相关研究的基础材料。     

Culture of Piglet Intestinal Epithelial Cells in vitro and the Infectivity of Porcine Epidemic Diarrhea Virus on the Cells

The study was aimed to establish a simple in vitro culture system for piglet small intestinal epithelial cells,and to provide materials for researches on porcine epidemic diarrhea virus (PEDV).In this study,newborn piglets that did not eat colostrum were used as the initial donors,and the primary cells were separated in vitro by scraping the intestinal mucosa from the intestinal lumen and mechanical separation and dispersion.The piglet intestinal epithelial cells were purified using 0.1% trypsin differential digestion method.Compared the effects of newborn weak piglets and normal piglets as donors on the activity of primary intestinal epithelial cells.MTT method was used to compare the proliferation activity of primary cells of different generations.Immunofluorescence and Real-time RT-PCR were used to detect the infection and proliferation of PEDV strain CV777 in primary intestinal epithelial cells.The results showed that the isolation and culture method in vitro used in this study could obtain primary intestinal epithelial cells with good proliferation activity,with obvious S-type cell proliferation curves.The cells with high purity and single morphology could be obtained by differential digestion,and had good proliferation activity after five consecutive passages.The proliferative activity of intestinal epithelial cells isolated from weak piglets and normal piglets had no obvious difference,and provided new way for reducing the cost of primary cell culture.The results of immunofluorescence and Real-time RT-PCR showed that PEDV could infect the primary intestinal epithelial cells,and replicated and proliferated in them.In this study,we established a simple,practical and low-cost method for in vitro culture of piglet primary small intestinal epithelial cells.The primary cells cultured by this method could be used as basic materials for the isolation and culture of PEDV and related research.     

In vitro cultivation and partial characterization of Lawsonia intracellularis from a Japanese field case of porcine proliferative enteropathy

Lawsonia intracellularis isolated from a Japanese field case of porcine proliferative enteropathy (PPE) was cultivated and partially characterized. The bacterial cells isolated from the intestinal mucosa of a pig suffering from the acute form of PPE were used to inoculate rat small intestine cells (IEC-18) and human epithelial cells (HEp-2). Infected foci, which were stained with L. intracellularis-specific rabbit antiserum, were observed in the cell culture at 5 days post inoculation. The DNA sequence of several genes in the Japanese isolate had high similarity with those of the L. intracellularis type strain, suggesting the genetically close relationship of the two strains. This is the first report describing the cultivation and partial characterization of L. intracellularis originated in Japan.     

一株仔猪屎肠球菌的分离及其种属鉴定

为分离出可用作猪用微生态制剂的肠道益生菌株,本研究选择锦州市凌海大业乡生态猪场为试验采样点,采集10日龄左右健康仔猪肠道粪样12份。利用MRS琼脂培养基分离培养初筛目标分离菌株的形态特征和培养特性,再通过生理生化鉴定法和16SrRNA基因分子鉴定法,最终确定1株分离菌为屎肠球菌,并命名为LN／EFl312株。为后续肠道益生球菌的益生功能研究奠定了物质基础。     

Variation in the immuno-pathological responses of lambs after experimental infection with different strains of Mycobacterium avium subsp. paratuberculosis

Ruminant infection by Mycobacterium avium subsp. paratuberculosis (MAP) causes a granulomatous inflammatory response in the intestine and associated lymph nodes. Differences either in the affected organs or in the inflammatory infiltrate were observed between species and individuals. Such differences are usually attributed to variations in host immune responses or to inconsistent effects among different MAP strains. To evaluate if different MAP strains induce different immuno-pathological responses in lambs, 28 one-month-old individuals were divided into six groups and inoculated with different MAP strains. Groups 1 and 2 were inoculated with two bovine strains isolated in Argentina that showed different genetic patterns after BstEII-IS900-RFLP (hereafter strains E and A respectively). Group 3 was inoculated with a bovine strain isolated in Spain obtained after a previous step of culture (patterns C1). Group 4 was inoculated with a homogenate of intestinal mucosa of a clinical case affected by the same bovine strain as that of group 3. Group 5 was inoculated with an ovine strain that was directly purified from the intestinal mucosa of a clinical case, and group 6 was kept as control (i.e. no inoculation). Peripheral immune responses were assessed until 150 days post-infection (dpi), when lambs were humanely killed. Pathological studies were performed in tissues from the intestine and lymph nodes. Lesion types and inflammatory infiltrates were examined as indicators of pathogenicity. All the lambs infected with bovine MAP strains showed a common lesion pattern regardless of the strain type. Such pattern was characterized by focal lesions mainly in the mesenteric lymph nodes, the presence of fibrous tissue, and, occasionally, necrosis in the granulomas as well as the presence of numerous giant cells. Differences in lesion severity were observed among groups: lambs from groups 1 and 2 had the highest number of granulomas and the largest lymph node area affected. Lesions in animals from group 5 (infected with an ovine strain) were more severe and occurred mostly in the intestinal lymphoid tissue; necrosis, fibrosis or giant cells were never detected in this group. These results indicate that the MAP strain type induces different pathological responses in lambs.     

LLO对大鼠肠黏膜微血管内皮细胞分泌NO和ET-1的影响

为探讨李斯特菌溶血素(LLO)导致大鼠肠黏膜微血管内皮细胞(RIMVECs)损伤的作用机制,将体外培养的RIMVECs分为对照组和LLO组,观察细胞形态变化,测定细胞生长情况,并检测细胞培养上清液中NO、ET-1浓度变化。结果表明:LLO组RIMVECs的细胞间隙变大,有大量细胞碎片漂浮;当LLO浓度达到50ng/m L以上时,测得细胞增殖(OD值)均呈极显著下降(P0.01);12 h内,NO、ET-1分泌量高于正常水平。试验表明,LLO引起RIMVECs细胞因子NO、ET-1分泌量升高,导致细胞微环境紊乱,是LLO导致肠黏膜微血管内皮细胞损伤的机制之一。     

1株猪源肠道益生菌的分离鉴定及其生物学特性研究

从健康猪肠道中分离出1株对仔猪大肠杆菌病和副伤寒病病原有较强拮抗作用的菌株,命名为W-02,对其进行菌落形态、生理生化特性和16S rDNA系统发育分析,确定该菌株为凝结芽孢杆菌。对该菌株胃肠道耐受特性和安全性进行了研究,结果表明,W-02菌株在人工胃液、人工肠液、猪胆盐和80℃高温表现出很强的耐受能力,急性毒性试验结果表明该菌株为无毒菌株。     

哺乳仔猪断奶前后肠道微生物培养组学研究

近年来国内外研究人员通过下一代测序和分离培养技术,结合宏基因组学相关的生物信息学手段证实了猪肠道微生物的组成及其代谢物活性对宿主的健康具有重要作用.受限于传统培养方法,猪肠道中大多数细菌仍无法培养,因此,为了突破研究人员研究宿主-菌株相互关系及益生菌株开发应用的限制,本研究对仔猪断奶前后回肠和结肠内容物微生物进行高通量...     

The experimental infection of piglets with a porcine reovirus.

A strain (Quebec) of reovirus isolated from the faeces of a pig with dysentery was neutralised by reovirus type 1 antiserum. Four of eight hysterectomy-produced, colostrum-deprived (HPCD) piglets dosed orally with the third cell culture passage of the virus developed diarrhoea and showed focal areas of villous atrophy in the small intestine. The virus was isolated from the intestinal tract of all eight specific pathogen free piglets, but not from three control animals. Nine germ-free piglets dosed orally with the eight cell culture passage of the virus showed neither clinical signs nor lesions, but virus was recovered from their intestinal tracts for 14 days after infection. No virus was isolated from four control germ-free piglets.