大熊猫小肠结肠炎耶尔森氏菌感染及治疗

大熊猫小肠结肠炎耶尔森氏菌感染及治疗叶志勇吕文其（中国成都大熊猫繁育研究基地,四川成都６１００８１）刘新华（重庆动物园,６３００５０）姜文球权洙浣金银规（韩国自然农园动物园）李昌雨（韩国汉城兽医大学）大熊猫消化系统疾病是一种发病率高、危害性大的常见病...     

鱼源小肠结肠炎耶尔森氏菌的耐药性研究

本试验采用聚合酶链反应(PCR)方法检测鱼源小肠结肠炎耶尔森氏菌的四环素类耐药基因(tetA、tetC、tetM)、磺胺类耐药基因(sul1、sul2、sul3)和氨基糖苷类耐药基因(aph(3')-Ⅲa、aac(6')-Ⅰb、ant(3')-Ⅰa、aac(3)-Ⅱa),并采用κ-B法检测该菌对14种抗生素的耐药表型。结果表明,小肠结肠炎耶尔森氏菌8种耐药基因(tetC、tetM、sul1、sul2,aph(3')-Ⅱa、aac(6')-Ⅰb、ant(3')-Ⅰa、aac(3)-Ⅱa)被检测出,而tetA、sul3基因未被检测出。该菌株对复方新诺明、磺胺异恶唑、红霉素、利福平、洁霉素和头孢噻吩6种抗生素耐药,对氟哌酸、氟苯尼考和头孢曲松等8种抗生素敏感。这为治疗小肠结肠炎耶尔森氏菌引起的鱼病积累了资料。     

羚牛等草食兽小肠结肠炎耶尔森氏菌病的诊断与治疗

羚牛，驯鹿等草食兽５种１０只选后发生急性出血性肠炎，经临床剖检，细菌学检查，生化反应综合诊断为由小肠结肠炎耶尔森氏菌引起的出血性肠炎，并以药敏试验为依据采取综合措施治愈９只，死亡１只。     

西宁野生动物园灵长类动物粪便中小肠结肠炎耶尔森氏菌的检测

应用细菌常规病原分离鉴定技术对西宁市野生动物园无菌采集的部分灵长类动物粪便进行小肠结肠炎耶尔森氏菌株的带菌率及其耐药率进行了检测.结果显示,在34份样品中共检出5株小肠结肠耶尔森氏菌,检出率为14.7％ (5/34);药敏试验结果表明,5株分离菌株对36种药敏纸片的平均耐药率为24.44％;致病性试验表明,4株分离菌对小鼠具有致死性效应,1株分离菌对小鼠有一定致病性.     

黄颡鱼感染小肠结肠炎耶尔森氏菌的组织病理学观察

为探讨小肠结肠炎耶尔森氏菌GM2402株对黄颡鱼的致病性,本试验采用人工回感试验测定其半数致死浓度(LC₅₀),同时采用组织病理学方法探讨该菌对黄颡鱼组织的影响.结果表明:回感18 h后,发病黄颡鱼腹部肿大,胸鳍、腹鳍基部出血,肛门红肿,其LC₅₀为3.0×10^6.2 CFU/mL.组织病理学观察肝细胞肿大、排列紊乱、广泛空泡变性;肾小球萎缩,肾小管上皮细胞肿大;脾脏组织结构疏松充满大量红细胞;鳃小片轻度水肿充血.超微结构观察结果显示肝脏和肠道细胞内线粒体均出现不同程度的肿胀,嵴断裂囊泡化.本试验结果表明小肠结肠炎耶尔森氏菌GM2402株对黄颡鱼有较强的致病性,可导致鱼体多个组织出现病变.     

猪肉中小肠结肠炎耶尔森氏菌失活／存活预测模型及控制研究

目的：建立小肠结肠炎耶尔森氏菌在猪肉中的存活／失活预测模型．并提出其控制措施。方法：用浓度梯度稀释法计细菌总量．按对数法作出不同条件下小肠结肠炎耶尔森氏菌的生长曲线。采用预测微生物学的基本方法和程序，并用CurveExpert1．38软件作为辅助工具。对试验数据进行拟合。结果：50℃、-18℃及速冻条件下该菌的初始菌数与时间为T时的菌数之比关于时间T的存活／失活预测模型符合Linear关系式。结论：实验表明，本菌对高温非常敏感，在冷冻．速冻过程中仍能存活。     

仔猪粪便中小肠结肠炎耶尔森菌的检测

应用常规细菌分离培养鉴定技术对西宁市某猪场断奶仔猪粪便中小肠结肠炎耶尔森菌带菌率进行了调查。结果显示,在60头断奶仔猪粪便中共检出7株小肠结肠炎耶尔森菌,阳性率为11.7%(7/60)。致病性试验表明有3株分离菌对小鼠具有致死效应。     

Yersinia enterocolitica infection in goat. A serological and bacteriological investigation

The distribution of agglutinating antibodies to Yersinia enterocolitica type 2 in sera from goats in an infected herd, and in 190 other animals from different parts of the country was studied. The faecal excretion of Yersinia enterocolitica from the same animals was also examined. Experimental inoculations of two goats were carried out. The serological results indicate that subclinical cases had occurred in the infected herd during the enzooty and during the following months. Faecal excretion of the organism was observed during the first month after the acute phase of the disease. It was found to be difficult to induce the disease experimentally, but clinical signs were observed in 1 goat after injection of live Yersinia enterocolitica intraperitoneally. The Widal-titre rose to 1/1250 during the first 2 weeks after the inoculation and then fell to 1/80 during the next 2 months. The serological results indicate infections with Yersinia enterocolitica, if a Widal-titre of at least 1/80–1/160 in the first month after the acute phase of a disease and a fall to a low level during the following months are found. Among animals from different partst of the country 16.3 % had a Widal-titre which indicated infection with Yersinia enterocolitica during the previous 3–6 months.     

Multiplex PCR method for differentiating highly pathogenic Yersinia enterocolitica and low pathogenic Yersinia enterocolitica,and Yersinia pseudotuberculosis

A multiplex PCR method for rapid and sensitive diagnosis, differentiating three pathogenic Yersinia groups such as the highly pathogenic Y. enterocolitica, including serotype O8, low pathogenic Y. enterocolitica, and Y. pseudotuberculosis, was developed. Four primer pairs were chosen to detect the genes fyuA, ail, inv, and virF, responsible for the virulence in pathogenic Yersinia species. Under the multiplex PCR conditions, the unique band patterns for the highly pathogenic Y. enterocolitica, low pathogenic Y. enterocolitica, and Y. pseudotuberculosis were generated from Yersinia strains. The detection limit of this method was 10¹–10³ CFU per reaction tube. This multiplex PCR method could detect highly pathogenic Y. enterocolitica O8 from the wild rodent fecal samples that were culture-positive. Therefore, the new multiplex PCR method developed in this study is a useful tool for rapid and sensitive diagnosis, distinguishing three pathogenic Yersinia groups.     

强毒小肠结肠耶氏菌生物素标记基因探针的研究

在构建的小肠结肠耶氏菌毒性质粒DNA基因文库pYB1～8与pYP1～6的基础上,筛选出了pYB7和pYP6克隆株.用限制性内切酶Bam HI消化pYB7,Pst消化pYP6,可分离出3.8kb和6.4kb的插入性DNA片段.以这两个基因片段为目的基因,用生物素化dUTP和光敏生物素标记,获得了生物素标记的基因探针.该探针能检出10pg以上的强毒小肠结肠耶氏菌DNA,不与无毒小肠结肠耶氏菌及大肠杆菌、鼠伤寒沙门氏菌、金黄色葡萄球菌等18种对照菌反应,具有高度的特异性和敏感性.pYB7与pYP6探针对不同血清型及来源的小肠结肠耶氏菌检测,其结果与自凝性试验、依钙试验等结果相符;对小肠结肠耶氏菌强毒株与无毒株检定的准确率为100%.     

小肠结肠耶氏菌O:9血清型与布氏杆菌M_5株共同抗原的免疫印迹分析

应用SDS-聚丙烯酰胺凝胶电泳和免疫印迹技术,对经超声波打碎的小肠结肠耶氏菌O:9血清型和布氏杆菌M5株全菌体蛋白成分进行了分子量测定及抗原分析。结果表明,小肠结肠耶氏菌O:9血清型Y15株与布氏杆菌M5株存在一条发生交叉反应的蛋白质共同抗原带,其分子量为11400。Y15株与M5株有多个分子量相同的条带,但不发生交叉反应。Y15株与M5株皆有多个各自特异的条带。     

腹泻水貂检出携带耶尔森菌HPI毒力岛的大肠杆菌

为了解大肠杆菌引起水貂腹泻的机理,进行了小肠结肠炎耶尔森菌HPI毒力岛基因的检测,并对其菌株做毒力试验。用PCR扩增法检测毒力岛基因irp2和fyua,小鼠腹腔注射检测菌株毒力。结果：从3个貂场腹泻病死水貂脏器以及粪便中分离出血清型分别为078、029和038的大肠杆菌,对3个血清型大肠杆菌进行毒力岛检测,均检出携带小肠结肠炎耶尔森菌HPI毒力岛基因irp2和fyua。3个血清型078、029和038的大肠杆菌均使小鼠发病死亡。结果表明水貂腹泻是由携带小肠结肠炎耶尔森茵HPI毒力岛基因irp2和fyua的大肠杆菌引起,该茵对水貂的健康具有潜在的威胁。     

感染小肠结肠炎耶尔森菌的黄颡鱼脾脏和肾脏组织病理学观察

为了解小肠结肠炎耶尔森菌感染黄颡鱼后对其脾脏和肾脏免疫器官结构的影响,通过人工感染小肠结肠炎耶尔森菌致病后,观察黄颡鱼脾脏和肾脏显微和超微结构的病理变化。结果显示,发病黄颡鱼腹部肿大,剖检可见脾脏、肾脏肿大;肾小球萎缩,肾小管上皮细胞水肿、变性;脾组织内充满大量红细胞;脾脏、肾脏细胞中线粒体均出现不同程度的肿胀,嵴断裂,囊泡化。这表明该菌对黄颡鱼具有明显的致病性,可导致其脾脏和肾脏发生严重的病变。     

Yersinia enterocolitica. Experimental pathogenicity for chinchilla.

An intragastric inoculation of approx. 2 × 10¹⁰ Yersinia enterocolitica cells killed chinchillas in three days in the case of four strains out of six tested. Because of the sensitivity of chinchillas to this bacterium, the test is useful for the evaluation of the virulence and invasiveness of Y. enterocolitica isolates. This animal model could also be used for studies on the mechanism of the infection.     

小肠结肠耶氏菌毒性质粒基因文库的构建

本研究以pAT153质粒为载体,构建了小肠结肠耶氏菌毒性质粒(pVYE)经限制性内切酶Bam HI和PstI消化产生的DNA片段的基因文库.实验克隆了8株(pYBI～8)pVYE的BamHI片段和6株(pYPI～6)pVYE的PstI片段的重组子.其中pYB4、pYB7、pYB8重组质粒中插入的pVYE—Bam HIDNA片段的分子量分别为8.7、3.8、20kb;pYP3、pYP5、pYP6重组质粒中插入的pVYE-Pst I DNA片段的分子量分别为4.5、3.0、7.0kb.本实验结果为进一步筛选和制备特异性基因探针奠定了基础.     

A long-term observation for ecology of pathogenic Yersinia in wild rodents living in Fukushima Prefecture,Japan

From 2012 to 2021, prevalence of pathogenic Yersinia in wild rodents captured in Fukushima Prefecture, Japan was investigated twice a year to clarify the ecology of this pathogen in wild rodent populations. Pathogenic Yersinia enterocolitica O8 was isolated from 13 (1.7%) of 755 wild rodents. The Y. enterocolitica O8 isolates harbored three virulent genes (ail, fyuA, and virF). This pathogen was isolated repeatedly from wild rodents in April 2015, 2016, and 2017, in June and November 2020, and in April 2021, which was 6 of 19 times of observations. All Y. enterocolitica O8 isolates showed the same PFGE patterns. These results indicated that the same clone of pathogenic Y. enterocolitica O8 has been maintained in wild rodent populations in Fukushima Prefecture. Therefore, wild rodent populations contribute substantially to the continuous transmission of Y. enterocolitica O8 and its persistence in the ecosystem. This is the first report on the isolation of pathogenic Y. enterocolitica O8 in wild rodents in Fukushima Prefecture, Japan.