鸡源致病性沙门氏菌的分离鉴定及血清型和药物敏感性分析

为了解当前广西鸡源致病性沙门氏菌的优势流行菌株及耐药情况,掌握其最新流行特点,本试验采集疑似病鸡的组织病料,对沙门氏菌进行增菌和分离培养,运用VITEK System ATB Expression法ID 32E肠杆菌鉴定试剂条对分离菌株进行生化试验鉴定,应用玻片凝集试验进行血清型鉴定,采用ATB VET药敏试剂条对30种肠杆菌抗菌药物进行药物敏感性试验。结果显示,从310份疑似鸡病料中分离鉴定出34株致病性沙门氏菌;这些分离菌株中有A群1株、C2群1株、B群15株和D群14株,另有3株未定型,其中以B群鼠伤寒沙门氏菌和D群鸡白痢沙门氏菌为优势菌株;34株分离株均对美洛培南、亚胺培南、大观霉素和安普霉素敏感,对氯霉素、卡那霉素、庆大霉素的耐药率小于10%,对阿莫西林、链霉素、氟甲喹、噁喹酸、磺胺甲噁唑、四环素和呋喃妥因耐药率介于50%和90%之间,而对青霉素、苯唑西林、夫西地酸、利福平和甲硝唑的耐药率达到100%,并且存在多重耐药现象。结果表明,鸡源致病性沙门氏菌的流行菌株具有一定的地域性,当前广西以鼠伤寒沙门氏菌和鸡白痢沙门氏菌为主,流行菌株耐药性严重,应高度关注。     

鸡源致病性沙门氏菌的分离鉴定及耐药性分析

通过对临床病死雏鸡的肝和心血处进行采样、分离、鉴定,获得13株致病性沙门氏菌。采用Kirby-Bauer法检测13株分离菌对10种抗菌药物的敏感性,结果表明,细菌对10种临床常用抗菌药物均表现出较高的耐药性,且均为两重或两重以上耐药,有的甚至达到对其中8种药物产生耐药性。13株分离菌对四环素的耐药率已高达100%,对复合磺胺、氧氟沙星、诺氟沙星、环丙沙星、氨苄西林、头孢唑啉和氯霉素的耐药率分别为92.3%、84.6%、84.6%、84.6%、69.2%、23.1%和7.7%;对庆大霉素和卡那霉素敏感。结果表明,沙门氏菌的耐药现象已十分严重。     

山东省鸡源沙门氏菌的分离鉴定及毒力基因分析

本研究旨在调查山东地区鸡源沙门氏菌的流行情况及毒力基因分布。在山东地区规模化养鸡场采集病料,利用四硫磺酸钠煌绿增菌液（TTB）进行沙门氏菌选择增菌,并对分离菌株进行纯化培养、革兰氏染色、生化试验、培养基菌落形态鉴定,利用沙门氏菌属及血清型特异性引物对分离菌进行PCR扩增鉴定,并利用PCR方法检测分离菌中33种毒力基因分布。结果显示,分离菌在LB固体培养基和麦康凯培养基上呈无色半透明、光滑且边缘整齐的菌落,镜检为革兰氏阴性、两端钝圆的小杆菌。生化试验、培养基菌落形态鉴定、沙门氏菌属及血清型特异性引物PCR扩增结果显示,共分离到25株鸡源沙门氏菌,其中肠炎沙门氏菌9株,鸡白痢沙门氏菌和鼠伤寒沙门氏菌各8株。毒力基因检测结果显示,33种毒力基因在25株分离菌中均有检出,其中20种毒力岛基因（invJ、virK、sopA、mogA、hilA、hisJ、ssaB、ssaQ、ssiD、misL、mgtC、orf319、siiE、siiD、bcfA、pipC、sopB、araB、spoB和avrA基因）、肠毒素基因（stn基因）及菌毛毒力基因（fimA基因）检出率均为100%;2种毒力岛基因（sipA和sodC基因）及4种毒力质粒基因（spvA、spvC、spvD和spvR基因）检出率均为96%,spoE基因检出率为68%,fliC基因检出率为36%,gipA与spvB基因检出率均为32%,sseL基因检出率为4%;25株鸡源沙门氏菌中,分别携带31种（8株）、30种（1株）、29种（15株）和24种（1株）毒力基因;毒力基因gipA和spvB只在鼠伤寒沙门氏菌中检出,且在鼠伤寒沙门氏菌中检出率为100%。本研究结果表明,山东地区鸡源沙门氏菌主要为肠炎沙门氏菌、鸡白痢沙门氏菌和鼠伤寒沙门氏菌,分离菌皆携带多种毒力基因,其中gipA和spvB毒力基因只在鼠伤寒沙门氏菌中检出,可作为鼠伤寒沙门氏菌鉴定的备选基因之一。     

西藏绵羊溶血性曼氏杆菌分离鉴定及药敏特性研究

为探究西藏自治区山南市某地农户饲养的绵羊大批死亡的原因,采集病死羊心脏、脾脏、小肠、肺脏、气管等病料,进行细菌培养、分离鉴定、PCR检测、血清型鉴定、毒力基因检测及药敏特性分析。结果显示:从病死羊肺脏病料中分离出一株溶血性曼氏杆菌,血清型为2型;特异性PCR检测为阳性,证明分离菌株为溶血性曼氏杆菌;药敏试验结果显示,分离的溶血性曼氏杆菌对大多数临床常用抗生素敏感;主要毒力基因lktA检测为阳性。该研究首次从西藏绵羊分离出溶血性曼氏杆菌,并研究了其药敏特性,为该病的诊断与治疗提供了参考。     

鸡源绿脓杆菌的分离鉴定及药敏试验

为确诊引起秦皇岛某鸡场细菌性疾病的主要病原菌,分析其耐药情况,为临床用药和治疗提供依据,本试验对送检的病死鸡肝脏中分离得到的4株菌进行形态特征观察、生化鉴定、免疫荧光染色观察、细菌16S rRNA基因PCR扩增与测序分析及药敏试验.结果显示,4株分离菌均为革兰氏阴性、带绿色荧光的短杆菌,PCR扩增出的特异性条带与预期大小相吻合,确定为绿脓杆菌.药敏试验结果显示,分离株对庆大霉素、阿米卡星、大观霉素、头孢他啶、头孢曲松、左氧氟沙星、恩诺沙星、氟苯尼考、多黏菌素B和磷霉素高度敏感,对四环素和强力霉素中度敏感,而对新霉素、头孢噻肟、头孢噻吩、阿莫西林、氨苄西林、红霉素、复方新诺明和磺胺甲氧异唑耐药.提示,该分离菌对常用抗生素耐药情况严重,养殖场应根据药敏试验结果选择敏感药物进行合理用药.     

山羊败血性链球菌的分离鉴定与药敏实验

通过临诊观察、病理剖检和细菌分离鉴定,确诊1例急性山羊败血性链球菌病。从患病羊心血分离的致病菌为革兰氏阳性菌,镜下呈单个球状或链状排列,在鲜血琼脂平板上呈β溶血,对头孢哌酮、头孢呋辛、米诺环素、环丙沙星、左氧氟沙星、氧氟沙星、诺氟沙星、呋喃妥因等抗菌药物高度敏感,可用于山羊链球菌病的治疗。     

宠物犬源沙门菌的分离鉴定及药敏试验

为分析洛阳地区宠物犬源沙门菌耐药情况,对采集的120份宠物犬肛门拭子样品进行沙门菌的分离鉴定,通过培养特性、革兰染色、生化试验、血清型检测和分子鉴定最终获得21株沙门菌。通过纸片扩散法进行23种抗菌药物敏感性试验,采用PCR检测四环素类、β-内酰胺类和磺胺类药物耐药基因共12种。药敏试验结果表明,21株沙门菌对头孢吡肟、诺氟沙星等6种药物均未表现耐药,对阿莫西林、磺胺异恶唑等17种抗菌药物表现不同程度的耐药,并出现多重耐药现象。PCR检测结果证实,耐药基因tetE、blaPSE、blaTEM、sulⅠ和sulⅡ的检出率最高。研究结果显示,洛阳地区宠物犬沙门菌的耐药情况较为严重,为防治宠物犬源沙门菌病及宠物犬临床用药提供参考。     

Isolation,Identification and Drug Susceptibility Test of Pseudomonas aeruginosa in Chickens

In order to detect the main bacterial pathogens and analyze the drug resistance for providing therapeutical guidance for drug administration,4 stains were isolated from death chickens of a breeding chicken farm in Qinhuangdao area.All 4 isolates were identified by morphological characteristics,biochemical test,immunofluorescent staining,amplification of 16S rRNA genes by PCR,sequence analysis and drug susceptibility test.The results showed that all 4 isolates were gram-negative,bacillus brevis with green fluorescent,and the sequence of 16S rRNA gene of isolates were analyzed and test results showed that specific amplified band was consistent with the expected size.The isolated bacteria were identified to be Pseudomonas aeruginosa.Results of drug susceptibility test indicated that the bacterial isolates were highly sensitive to gentamycin,amikacin,spectinomycin,ceftazidine,ceftriaxone,levofloxacin,enrofloxacin,florfenicol,polymyxin B and fosfomycin,intermidiate sensitive to tetracycline and doxycyclin,resistant to neomycin,cefotaxime,cephalothin,amoxicillin,ampicillin,erythromycin,trimethoprim-sulfamethoxazole and oxygen sulfamethoxazole isoxazole.The results suggested that the resistances of Pseudomonas aeruginosa isolates to commonly used antibiotics were serious.So antibiotics should be reasonably selected based on the result of drug susceptibility test in this farm.     

安徽地区禽源沙门氏菌分离鉴定与耐药性研究

为了解安徽地区禽源沙门氏菌的流行菌株和耐药情况,本研究通过培养特性、镜检、生化试验、PCR和血清型鉴定等,对疑似发病禽的116份病料进行沙门氏菌分离鉴定,并采用Kindy-Bauer法对分离菌株进行耐药性分析。试验共分离到42株禽源沙门氏菌,其中包含5个大群11种血清型。B群30株、A群2株、D群7株、C₁群2株、C₂群1株,其中以B群鼠伤寒沙门氏菌为优势菌株。42株分离菌对阿米卡星、左氧氟沙星和环丙沙星的耐药率均低于20%,而对氨苄西林、链霉素、呋喃唑酮、大观霉素、复方新诺明和多西环素的耐药率达50%以上,其中对氨苄西林的耐药率达97.62%,且41株分离菌株存在多重耐药。结果表明,目前安徽地区禽源沙门氏菌优势流行株是鼠伤寒沙门氏菌,流行菌株呈现较严重的耐药性。本研究为指导临床用药及禽源沙门氏菌的综合防制提供数据支撑。     

宁夏部分地区鸡大肠埃希菌的分离与血清型鉴定

为调查宁夏地区鸡大肠杆菌病的流行情况,采集宁夏4个地区的103个规模化鸡场疑似病料151份,通过病原菌分离鉴定及生化试验,鉴定出鸡大肠埃希菌109株,平均分离率为72.19%(109/151);动物试验证实,59株分离菌有致病性,占分离菌54.13%(59/109);对致病性菌株进行血清型鉴定,确定血清型种类13种38株,占致病性菌株64.41%(38/59);优势血清型分别为O15、O18、O78、O88,占定型菌株63.16%(24/38)。     

Isolation,Identification and Drug Resistance Analysis of Avian Salmonella in Anhui

In order to investigate the situation of predominant strain and antibiotic resistance of avian Salmonella in Anhui province, 42 strains Salmonella were isolated and identified from 116 suspected samples by culture characteristics, microscopic examination, biochemical test, PCR and serological identification,then drug resistance was analyzed by Kindy-Bauer method. There were 5 groups of 11 serotypes in 42 strains Salmonella, of which 30 strains belonged to B serogroup, 2 strains to A serogroup, 7 strains to D serogroup, 2 strains to C₁ serogroup, 1 strain to C₂ serogroup, and Salmonella Typhimurium of B serogroup were the predominant serotypes. All Salmonella isolates resistant rates to amikacin, levofloxacin and ciprofloxacin were less than 20%, and the resistance rates to ampicillin, streptomycin, furazolidone, spectinomycin, cotrimoxazole and doxycycline were more than 50%, while resistance rate to ampicillin was 97.62%, and 41 isolates had multiple drug resistance. The results showed that Salmonella Typhimurium was the predominant serotypes of avian Salmonella in Anhui, and isolates showed multiple drug resistance. This study would provide the data basis for the comprehensive prevention and treatment of clinical medicine of avian Salmonella.     

乌鲁木齐某农场沙门氏菌的分离鉴定

新疆乌鲁木齐某养鸡场近期发生一起临床表现为实验结果可为精神萎顿、食欲减少、腹泻、有的出现眼盲或肢关节呈跛行等症状的疫情。对送检病料进行病原分离和生化鉴定,结合分子生物学方法、动物回归及药敏试验,确认该鸡场疫情病原为鸡白痢沙门氏菌。建议加强饲养管理、改善环境卫生、做好隔离消毒。本实验可为该病的防控提供参考。     

1株鸡源肠炎沙门氏菌的分离鉴定及致病性分析

【目的】确定云南某蛋鸡场送检的疑似沙门氏菌病病料中的主要病原菌种类,并测定其致病性及药物敏感性。【方法】通过细菌分离培养、形态观察、生化试验、血清学鉴定及分子学方法进行细菌分离鉴定;通过对鸡的致病性试验及毒力基因检测分析分离菌的致病性;通过药物敏感性试验检测分离菌耐药情况。【结果】分离菌株在沙门氏菌显色培养基上形成紫色菌落,镜检为革兰氏阴性短杆菌,疑为沙门氏菌,命名为S2;生化试验结果显示,菌株S2赖氨酸脱羧酶、硫化氢、卫矛醇试验结果均为阳性;靛基质、尿素酶、氰化钾、山梨醇、水杨苷、β-半乳糖苷、丙二酸盐生化试验结果均为阴性,判断菌株S2为典型沙门氏菌属;血清学鉴定结果显示,菌株S2属于D群肠炎沙门氏菌,其抗原式为1,9,12:g,m;遗传进化分析结果显示,菌株S2与肠炎沙门氏菌MT621365处于同一分支,自展值为99%;致病性试验结果表明,分离菌株对雏鸡有较强的致病性,感染6 d后雏鸡全部死亡(10/10);毒力基因检测结果显示,菌株S2携带invJ、hilA、ssaB、misL、mgtC、orf319、sopB、spvA、spvB、spvC、spvR、stn和fimA毒力基因,不携带sopA、fliC毒力基因;菌株S2对多数抗菌药敏感程度很高,对阿莫西林-克拉维酸、头孢西丁、四环素等12种抗菌药均敏感,对甲氧嘧啶、复方新诺明和林可霉素耐药。【结论】本研究分离到了1株肠炎沙门氏菌,其毒力较强,可造成雏鸡较高的死亡率,对大部分常用抗菌药敏感,对甲氧嘧啶、复方新诺明和林可霉素具有耐药性。本研究结果可为沙门氏菌病的临床诊断和药物有效性评价提供一定参考依据。     

河南省鸡源结肠弯曲杆菌的分离鉴定及耐药性分析

[目的]掌握河南省鸡源结肠弯曲杆菌的分布和对抗菌药物的耐药情况。[方法]从养殖场抽取鸡泄殖腔拭子460份进行结肠弯曲杆菌分离培养,采用PCR、基质辅助激光解吸电离飞行时间质谱进行鉴定,并通过微量肉汤稀释法对分离菌株进行药物敏感性分析。[结果]共分离结肠弯曲杆菌42株,分离率为9.13%;分离的结肠弯曲杆菌对环丙沙星、萘啶酸、阿奇霉素耐药严重,耐药率分别为95.24%、90.48%、71.43%。[结论]河南省鸡源结肠弯曲杆菌耐药情况较为严重,应规范使用抗菌药物。     

副猪嗜血杆菌的分离鉴定及药敏试验

黑龙江省近几年频繁发生以多发性浆膜炎、关节炎、脑膜炎以及急性死亡为特征的传染病,为猪场造成严重经济损失,为了确诊是否有副猪嗜血杆菌感染,采集病死猪肝脏、脾脏和肺脏等病料,进行副猪嗜血杆菌的分离;对其理化特性进行鉴定,应用PCR方法对其16S rRNA基因扩增后进行克隆测序,将测序结果在GenBank上进行BLAST分析,把相近的基因序列应用DNAStar软件进行同源性和进化关系分析;用分离菌株进行药物敏感性试验,筛选敏感药物。结果分离出一种具有多形性的NAD依赖性菌株,经鉴定为副猪嗜血杆菌;16S rRNA基因进化分析结果表明,分离菌株与以往报道的副猪嗜血杆菌中国、日本和美国分离株属于同一亚群,而西班牙分离株属于单独的亚群;分离菌株对壮观霉素和头孢拉定高度敏感。     

红斑丹毒丝菌的分离鉴定及药敏试验

为确定引起广东潮州某猪场后备母猪发病的病原,采用自制的含马血清的牛心浸汁替代琼脂培养基,从送检的肝脏组织病料中分离到菌落形态一致的1株细菌,根据其菌落形态特征、革兰染色观察、生化特性和16SrDNA序列分析,最终鉴定为红斑丹毒丝菌,将其命名为CZ1。CZ1对31种常用抗菌药物的药敏试验结果显示,该菌对青霉素类和头孢菌素类、四环素类、呋喃类、大部分氨基糖苷类、大环内酯类、喹诺酮类均敏感;对磺胺类药物、利福平和氟罗沙星耐药。对菌株CZ1和实验室早前分离的韶关株SG7分别进行小鼠LD50测定,测得CZ1株和SG7株的LD50分别为1.02×10^4 CFU/mL和7.3×10^3 CFU/mL,表明CZ1株的毒力弱于韶关分离株SG7株。用大鼠分别制备抗CZ1株和SG7株高免血清,血清玻片凝集试验显示,抗CZ1株和SG7株高免血清均能与自身菌株新鲜培养物发生强的凝集反应。交互凝集试验显示,抗CZ1株高免血清能与SG7株新鲜培养物发生强的凝集反应,抗SG7株高免血清能与CZ1株新鲜培养物发生强的凝集反应,表明此次潮州分离株CZ1与SG7株很有可能属于同一种血清型分离株。而2种抗血清均不与现有疫苗株GC42发生凝集反应,提示猪丹毒病的病原血清型可能发生了变化。     

腹泻仔猪肠道致病性大肠埃希氏菌和沙门氏菌的分离鉴定与耐药性研究

为探明仔猪细菌性腹泻肠道致病性大肠埃希氏菌和沙门氏菌流行的血清型、耐药表型及耐药基因型,本试验采集了贵州省5个地（州）市7个规模化养猪场的128份腹泻仔猪肠道样本,并对采集的样本进行了大肠埃希氏菌和沙门氏菌分离与鉴定,通过动物试验鉴定菌株的致病性,利用血清学方法鉴定其血清型,并通过药敏纸片法对主要致病菌进行耐药性研究,采用PCR技术检测各致病菌株耐药相关基因,分析细菌耐药表型和耐药基因型相关性。结果显示,本研究共分离鉴定到78株致病性大肠埃希氏菌与21株沙门氏菌,致病性大肠埃希氏菌血清型以O138、O87为主,沙门氏菌血清型以鼠伤寒沙门氏菌、肠炎沙门氏菌居多;药敏试验结果表明,本试验分离到的78株致病性大肠埃希氏菌对β-内酰胺类药物耐药率达80%以上,对其他种类的抗菌药耐药率均超过40%,分离鉴定的21株沙门氏菌对氨基糖苷类药物耐药率达50%以上,对其他种类的抗菌药耐药率均达20%以上;本试验分离鉴定的致病性大肠埃希氏菌共检出12种耐药相关基因,沙门氏菌共检出10种耐药相关基因,两种细菌耐药基因型与耐药表型符合率均达60%以上,且均为多重耐药。本研究为仔猪腹泻的综合防控提供了理论依据。     

Isolation,Identification and Drug Resistance Analysis of Interstinal Pathogenic Escherichia coli and Salmonella Isolated from Diarrhea Piglets

In order to find out the serotype, resistant phenotype and genotype of Escherichia coli (E. coli) and Salmonella in piglets, this study collected 128 samples of diarrhea piglets from seven large-scale pig farms in five cities in Guizhou province, and the E. coli and Salmonella were isolated and identified. The pathogenicity of the strain was identified by animal test. The drug resistance of the main pathogen was tested by drug susceptibility paper. The resistance gene of each pathogen was detected by PCR. The drug resistance and genotype correlation of the bacterial were analyzed. The results showed that 78 strains of pathogenic E. coli and 21 strains of Salmonella were isolated and identified in this study. The serotypes of pathogenic E. coli were predominantly O138 and O87. Salmonella Typhimurium and Salmonella Enteritidis were predominant serotypes. The susceptibility test showed that the resistant strains of 78 strains of E. coli were more than 80% resistant to β-lactams and more than 40% for other antibacterials. The resistance rate of 21 strains of Salmonella to aminoglycosides was more than 50% and more than 20% to other types of antibacterials; 12 and 10 kinds of drug resistance-related genes of E. coli and Salmonella were detected, respectively; The coincidence rate of resistant genotype and phenotype of two kinds of bacteria were above 60%, and both were multiple drug resistance. This study provided a theoretical basis for comprehensive prevention and control of piglets diarrhea.     

新疆地区2株驴源马流产沙门菌的分离鉴定及致病性分析

【目的】研究确定导致新疆地区2个规模化驴场出现流产的疑似病原沙门菌,并探究其致病能力和耐药性情况。【方法】通过细菌分离培养、形态学观察、生化试验对分离菌进行了鉴定,并对分离菌的鞭毛基因FliC进行了PCR扩增及序列分析,通过致病性测定、荷菌量检测及病理组织学观察,鉴定和分析了分离菌的致病性,并通过药敏试验分析其耐药性。【结果】通过细菌分离培养、形态学观察、生化试验,确定分离到的2株细菌均为马流产沙门菌,分别命名为G1-1和XD1-2。对这2个分离株的鞭毛基因FliC的遗传进化分析结果显示,2株分离菌FliC氨基酸序列之间的相似性为99.0%,2株分离菌FliC氨基酸序列与爱尔兰马源马流产沙门菌分离株Ireland-HE801373、Ireland-HE801378株相似性均最高,且均为99.3%;分离株G1-1与爱尔兰马源马流产沙门菌分离株Ireland-HE801373和Ireland-HE801378及国内马源马流产沙门菌分离株China-KJ486797.1和China-KJ486769.1亲缘关系较近,分离株XD1-2则与美国肠炎沙门菌分离株USA-EBQ1214032.1亲缘...     

Isolation,Identification and Analysis of Serotype and Drug Resistance of Salmonella from Pigeon in Shanghai

The purpose of this study was to investigate the molecular types and antimicrobial resistance of Salmonella isolated from pigeon in Shanghai from 2011 to 2014. A total of 92 fecal samples were collected from markets. XLD plate and Salmonella chromogenic medium were used to isolate suspected Salmonella colonies, and then determined the number of Salmonella through gram staining and biochemical tests. Among them, 24(26.1%) were positive for Salmonella. Kauffmann-White method and the K-B method were used respectively for serotype identification and susceptibility testing. In result, serological identification showed that 24 isolates from pigeon could be divided into 3 serotypes including S.typhimurium (66.7%), S.agona (25.0%) and S.corvallis (8.3%). Drug susceptibility test showed that 75.0% of the isolates were resistant to one antibiotic at least. The highest level resistance were found for tetracycline as well as sulfisoxazole (62.5%), followed by streptomycin (58.3%), nalidixic acid (50.0%) and ampicillin (20.8%). Multi-drug resistant strains accounted for 62.5% (15), in which the largest number of strains (7, 16.7%) were resistant to four drugs. In addition, isolates were 100.0% susceptible to cefepime, cefotaxime, ceftazidime, imipenem and ofloxacin, but 12.5% were moderately sensitive to the amoxicillin/clavulanic acid and ciprofloxacin. The study showed that Salmonella had a high separation rate of pigeon in Shanghai farmers market and performed serious multidrug resistance, which would bring great challenges and risks to the prevention and control of Salmonella in food, so that the prevalence and drug resistance of Salmonella in pigeon deserved close attention.