河北省保定地区雏鸡鸡白痢沙门菌的分离鉴定及耐药性检测

为了有效防控雏鸡白痢沙门菌病,试验自保定某蛋种鸡场疑似为鸡沙门菌病雏鸡采取肝脏病料进行细菌的分离培养、生化鉴定、鸡白痢沙门菌多价血清学鉴定及沙门菌inv A基因PCR鉴定,并对分离菌进行致病性检测及对7种抗菌药物的耐药性检测。结果表明,10株分离菌的形态特征、生化特性、血清学鉴定结果符合鸡白痢沙门菌的特征,说明从病死雏鸡组织中分离出的细菌为鸡白痢沙门菌。沙门菌inv A基因PCR扩增出373 bp目的片段,进一步证实分离菌为鸡沙门菌。用分离菌株接种雏鸡,其死亡率高达90%,表明该菌株有很高的致病性。10株分离菌株对阿莫西林和青霉素耐药率达100%,对阿齐霉素的耐药率达90%,而对阿米卡星、头孢唑林和庆大霉素耐药性较弱、对氧氟沙星较为敏感。3耐菌株多重耐药比例为42.86%,4耐菌株多重耐药比例为57.14%,5耐菌株多重耐药比例为71.43%,说明雏鸡沙门菌分离株对所试抗菌药物均表现出不同程度的耐药性,且表现出多重耐药现象。本试验为鸡白痢沙门菌病的防治提供参考。     

一株高致病性鸡伤寒沙门菌的分离与鉴定

对某蛋鸡场送检的10日龄左右病死雏鸡,进行了病理剖检,并开展了病原微生物分离培养、PCR鉴定、生化鉴定、血清学分型、动物回归、药物敏感性试验.结果显示,分离菌株为革兰氏阴性小杆菌,能够与鸡白痢鸡伤寒沙门菌阳性血清产生明显的凝集,PCR结果证实该分离株为鸡伤寒沙门菌;动物回归试验中,分离的鸡沙门菌具有较强的致病性;药物敏感性试验表明,分离株对替米考星、磷霉素、阿莫西林耐药,使用敏感药物氟苯尼考对鸡群沙门菌的控制取得了良好的治疗效果.     

同时感染雏鹅的两种血清型沙门菌分离鉴定及遗传进化分析

为了解鹅源沙门菌的病原学特性,本研究从一发病雏鹅体内分离到2株细菌,根据培养特性、染色镜检、生化试验、PCR检测及血清学分型结果鉴定,分离菌分别为鼠伤寒沙门菌与印第安纳沙门菌。药敏试验表明,2个沙门菌分离株均为多重耐药菌株,其中鼠伤寒沙门菌耐3种药物(耐药率16.7%),而印第安纳沙门菌耐12种药物(耐药率66.7%);2个分离株均对头孢曲松钠、呋喃唑酮、多粘菌素B及链霉素表现高度敏感。对分离菌株16S r DNA基因进行扩增测序后,与Gen Bank登录的22株不同血清型沙门菌序列进行比较,构建系统进化树,结果显示:2个分离菌株具有较高同源性,与大多数泛嗜性血清型亲缘性较近,而与鸡沙门菌感染密切相关的血清型亲缘性相距较远。本研究为鹅沙门菌病的流行病学调查积累了数据,也为临床防控提供了试验依据。     

鹅源鼠伤寒沙门菌的分离鉴定及药敏试验

对临床送检疑似鼠伤寒沙门菌感染的病例进行细菌分离,利用沙门菌所具有的种属特异性的亲膜蛋白基因inv A做PCR鉴定,结合生化鉴定和血清型鉴定,并分析分离菌的耐药性。从4例病死鹅病料中共分离到4株细菌,经inv A PCR鉴定、生化特点和血清型鉴定确认为鼠伤寒沙门菌。药敏试验结果显示,4株分离菌对庆大霉素、诺氟沙星、头孢曲松等药物敏感性较强,但对多黏菌素B、多西环素耐药。其中,有2个分离株表现出多重耐药性。     

鹅源鼠伤寒沙门茵的分离鉴定及遗传进化分析

从发病雏鹅肝脏中分离到1株细菌,根据培养特性、镜检结果、生化特征、血清学分型及PCR检测结果,表明分离茵为鼠伤寒沙门菌.动物试验结果表明,分离菌可致死雏鹅;药敏试验表明,该菌对环丙沙星、痢特灵、茵必治、阿米卡星、链霉素等药物敏感,对利福平和四环素耐药,与其它家禽或水禽来源的沙门菌相比,该分离株耐药性不严重.将分离茵16SrDNA基因进行扩增,PCR产物经胶回收试剂盒纯化后测序,序列Blast分析与血清型鉴定一致.将序列与GenBank上已登录的29株不同来源的鼠伤寒沙门茵序列进行比较,构建系统进化树,结果显示分离菌与美国分离的VDL-SS334株位置最为接近,同源性为95.9％.     

鸡大肠埃希菌与沙门菌混合感染的病原分离鉴定及药敏试验

对武汉市某鸡场送检的病鸡进行病理剖检,病理组织切片观察,并从病鸡的肝脏和心脏中分离得到两株细菌,依次进行分离纯化培养、镜检、生化鉴定及鸡白痢鸡伤寒玻片凝集试验,确定一株为大肠埃希菌,另一株为沙门菌.通过进行小鼠感染试验,表明所分离的两株细菌均有致病力.通过药敏试验分析它们对卡那霉素、恩诺沙星、利福平、庆大霉素、氟哌酸、...     

动物源性沙门菌的血清型、生物被膜形成能力和耐药性分析

本研究旨在探讨动物源性沙门菌的血清型分布、生物被膜形成能力及其耐药性.从不同动物病料中分离细菌,以PCR方法鉴定沙门菌,结合玻片凝集法和16S rRNA序列测定确定沙门菌的血清型和分布,结晶紫染色定量法检测分离株的生物被膜形成能力,药敏试验检测分离菌对常用药物的敏感性.结果共分离鉴定出58株沙门菌,包括鸡白痢沙门菌、鼠伤寒沙门菌、肠炎沙门菌、丙型副伤寒沙门菌、乙型副伤寒沙门菌、都柏林沙门菌和阿哥那沙门菌等7种血清型,其中鸡群以鸡白痢沙门菌感染为主,肠炎沙门菌次之;水禽以鼠伤寒沙门菌感染为主.生物被膜测定结果显示51.72％的沙门菌分离株可形成生物被膜,其中83.33％的鼠伤寒沙门菌可形成生物被膜.20种抗生素(氨基糖苷类、磺胺类、喹诺酮类、林可酰胺类、氯霉素类、青霉素类、四环素类和头孢菌素类)敏感性试验表明所有菌株对林可霉素耐药,51.72％对4种及其以上抗生素耐药,并出现了1株对所有受试抗生素均耐药的鼠伤寒沙门菌.结果表明鸡白痢沙门菌、鼠伤寒沙门菌和肠炎沙门菌是为目前在家禽中分离的优势血清型;同时具有生物被膜形成能力和多重耐药性的沙门菌将对家禽疾病控制和公共卫生带来更大的威胁.     

猪源鼠伤寒沙门菌的分离鉴定与耐药性分析

为确定江西2个规模养猪场患病死亡仔猪的病因,通过病理剖检、常规细菌学检查,从采集的病料中进行病原菌的分离鉴定,并利用动物试验和药敏试验,确定分离菌的致病性和敏感药物。结果表明,6株分离菌均符合鼠伤寒沙门菌的生物学特性,分离菌可在48h内致死小鼠,分离菌的耐药性较强,对阿米卡星、卡那霉素、头孢噻肟较敏感,对另外9种抗生素低敏或耐药。确定2个猪场仔猪发病的主要致病菌是鼠伤寒沙门菌。     

河南省鸡源沙门菌新近流行株的分离鉴定及其耐药性分析

旨在了解河南省鸡沙门菌临床分离株血清型分布及药物敏感情况。从河南省各地区养鸡场共采集了沙门菌可疑病料617份,按常规方法进行分离,经生化、PCR及血清型鉴定,共分离出98株鸡源沙门菌,其中血清型定型82株,主要为肠炎沙门菌(62.24%)、鸡白痢沙门菌(18.37%)。进一步Kindy-Bauer(KB)法检测分离菌株对22种药物敏感性表明,所有分离菌株对多粘菌素B、阿米卡星、庆大霉素、萘啶酸较为敏感,对阿齐霉素和氨苄西林耐药率在60.00%～70.00%之间,对红霉素和青霉素表现为极高的耐药性,耐药率高达100.00%。三重以上耐药菌株高达96.94%,七耐以上的菌株为56.12%,耐药最多的菌株可耐18种抗生素。由此可见,河南省禽类沙门菌的感染流行主要以肠炎沙门菌和鸡白痢沙门菌为主,多重耐药十分严重,严重影响了禽类沙门菌病的诊治与预防。     

15株禽源沙门菌的分离鉴定与耐药性试验

本试验旨在对沙门菌临床分离株进行鉴定及其耐药性进行研究。通过收集2012年2月-7月间具有典型沙门菌病病变特征的病例,无菌采集病死禽的肝脏、心、卵巢等脏器接种血清营养琼脂培养基,以麦康凯琼脂培养基纯化分离菌,并检测其生化特点,16S rDNA基因测序法快速鉴定分离菌株,然后进一步鉴定血清型;同时用15种抗菌药物进行药敏试验。结果显示,有15株细菌在麦康凯琼脂培养基上为无色、透明的菌落,能利用葡萄糖、甘露醇,不发酵乳糖、蔗糖,符合沙门菌的生化特性。同时,15株沙门菌的16S rDNA序列分析结果显示,分离株的同源性达到97%以上。15株沙门菌的血清学试验结果显示,有7株鼠伤寒沙门菌、5株伤寒沙门菌、2株肠炎沙门菌、1株鸡白痢沙门菌。药敏试验结果显示,分离细菌对链霉素、萘啶酸、强力霉素和美洛西林耐药率高。     

Drug resistance and biochemical characteristics of Salmonella from turkeys.

A study was conducted to determine the antibiotic resistance and biochemical characteristics of 2690 Salmonella strains belonging to 52 serovars and isolated from environmental and feed samples from 270 turkey flocks in Canada. Resistance of the Salmonella strains to the aminoglycoside antibiotics varied widely; none of the strains were resistant to amikacin, 14.2% were resistant to neomycin, 25.8% were resistant to gentamicin, and 27.7% of the strains were resistant to kanamycin. Most strains (97.6%) were resistant to the aminocyclitol, spectinomycin. Regarding resistance to the beta-lactam antibiotics, 14.3% and 14.4% of the strains were resistant to ampicillin and carbenicillin, respectively, whereas only 5 (0.2%) of the strains were resistant to cephalothin. None of the strains were resistant to the fluoroquinolone ciprofloxacin or to polymyxin B. Resistance to chloramphenicol and nitrofurantoin was found in 2.4% and 7% of the strains, respectively. Only 1.7% of the strains were resistant to the trimethoprimsulfamethoxazole combination, whereas 58.1% were resistant to sulfisoxazole. Thirty-eight percent of the strains were resistant to tetracycline. Salmonella serovars differed markedly in their drug resistance profiles. Biochemical characterization of the Salmonella showed that the S. anatum, S. saintpaul and S. reading serovars could be divided into distinct biotypes.     

猪链球菌Ⅱ型四川分离株对抗菌药的敏感性分析

对四川省不同地区链球菌病病死猪体内分离的7株链球菌Ⅱ型进行抗菌药敏感性试验,用链球菌兰氏D群C55914作为对照,比对菌为肺炎链球菌质控菌株ATCC49619.在32种抗菌药中,选择29种药物进行纸片法药敏试验.试验结果表明:7株链球菌耐药谱非常相似,对青霉素、氨苄西林、阿莫西林、奥格门丁、卡那霉素、红霉素、壮观霉素、氯霉素、氟苯尼考、头孢氨苄、头孢拉定、头孢他定、头孢呋辛、头孢曲松、环丙沙星、氧氟沙星、二氟沙星、沙拉沙星、达氟沙星、恩诺沙星、复方新诺明、万古霉素、亚安培南均敏感;对阿米卡星、新霉素、庆大霉素3株敏感,4株中敏;对四环素和多西环素2株中敏,5株耐药;对链霉素1株中敏,6株耐药.用21种药物进行最小抑菌浓度(MIC)测定,结果7株均对青霉素、氨苄西林、卡那霉素、红霉素、头孢他定、头孢呋辛、头孢曲松、头孢噻呋、环丙沙星、达氟沙星、氧氟沙星、二氟沙星、沙拉沙星、恩诺沙星、磺胺六甲氧嘧啶敏感,1株对新霉素和阿米卡星中敏,2株耐药;所有菌株对四环素、土霉素、多西环素、链霉素均耐药.两种方法总体结果一致.     

广东省部分地区禽源和猪源沙门菌血清型与耐药性及Ⅰ类整合子分析

为了解广东省禽场和猪场沙门菌的血清型和耐药性情况,本研究2015年从广东省多个养殖场共采集样品126份,分离出沙门菌24株,分离率为19.04%。采用Kauffmann-White法、Kirby-Bauer法和PCR方法对分离株进行了血清型鉴定、药敏试验和Ⅰ类整合子检测。血清型鉴定结果显示,24株沙门菌共鉴定出4种血清型,分别是鼠伤寒沙门菌(14株)、印第安纳沙门菌(8株)、科瓦利斯沙门菌(1株)和阿尔巴尼沙门菌(1株)。药敏试验结果显示,分离株对四环素(83.33%)、氨苄西林(70.83%)、磺胺异恶唑(70.83%)、萘啶酸(66.67%)、复方新诺明(58.33%)、卡那霉素(54.17%)、阿米卡星(54.17%)、庆大霉素(50.00%)的耐药率较高,有54.17%(13/24)的菌株对8种及8种以上抗菌药物耐药。Ⅰ类整合子检测结果显示,Ⅰ类整合酶阳性率为29.17%(7/24),整合酶阳性菌株中仅有1株扩增到携带耐药基因aadA2的基因盒。上述结果表明,广东省禽场和猪场沙门菌分离率较高,耐药情况严重,菌株的耐药性与其血清型和Ⅰ类整合子的携带有一定的相关性。     

A survey of antimicrobial resistance in Salmonellae isolated from animals in England and Wales during 1984-1987

Resistance to 14 antimicrobial substances was tested in 18,647 salmonella strains isolated from animals, their environment and from animal feeds during the period 1984-1987. Of the 2284 Salmonella dublin strains the percentage sensitive to all the antimicrobial substances ranged from 18.1 to 26.8. Resistance to the higher concentration of streptomycin (S25) ranged from 1.9 to 6.4%, whereas the corresponding figures when the lower concentration (S10) was used were 32.3 and 63.8%. Resistance to the higher sulphonamide concentration (Su500) never exceeded 3.3%, although in 1987 70.3% of strains showed resistance to the lower concentration. In general, less than 1% of strains showed resistance to the other antibacterial substances. No strains resistant to amikacin (AK), apramycin (Apr), gentamicin (CN) and colistin (CT) were detected. Of the 8677 S. typhimurium strains the percentage sensitive to all the antimicrobial agents ranged from 6.6 in 1985 to 13.6 in 1987. Resistance to tetracycline (T), ampicillin (PN), trimethoprim (TM) and chloramphenicol (C) ranged from 36.5 to 58.8%, the highest percentages being detected with tetracyclines. Less than 1% of strains showed resistance to furazolidone and none was resistant to amikacin and colistin. Resistance to apramycin ranged from 3.1% in 1984 to 11.6% in 1985; the figures for gentamicin were approximately half that of apramycin. In 1984, 41.6% of strains showed resistance to neomycin but only 8.5% in 1987. The fall in neomycin resistance was associated with the epidemic spread of the commonest phage-type DT204C becoming sensitive to neomycin. Of the 7687 strains of serotypes other than S. typhimurium and S. dublin the percentage sensitive to all antimicrobial agents ranged from 23.7 in 1985 to 14.7 in 1987. Resistance to tetracyclines and sulphonamides (Su500) ranged from 5.2 to 12.1% and 5.8 to 13.3% respectively. Resistance to the other antimicrobial agents was usually less than 5%.     

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.     

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。     

Salmonella Gallinarum field isolates and its relationship to vaccine strain SG9R

1. The aim of the present study was to determine if the 9R-strain of the Salmonella Gallinarum live vaccine was responsible for having fowl typhoid outbreaks in chicken flocks from both chicken and turkey breeders as well as to verify the antimicrobial resistance of the isolates from the outbreaks.

2. The triplex polymerase chain reaction, standard antimicrobial test, beta-lactamase genes identification and Ion Torrent PMG whole-genome sequence were used in the field isolates and in the vaccine strain of S. Gallinarum.

3. The 60 tested isolates were not from vaccine origin and manifested high resistance to drugs from macrolide and quinolone groups. Whole-genome sequencing (WGS) and single nucleotide polymorphism analysis on selected isolates for core genes from Salmonella enterica confirmed the wild origin of these isolates and showed two possible sources of S. Gallinarum in the studied outbreaks.

4. S. Gallinarum isolated from fowl typhoid outbreaks in the studied period were not caused by the use of the SG9R live vaccine. The source of strains sequenced was diverse.     

High prevalence of multiple resistance to antibiotics in Salmonella serovars isolated from a poultry slaughterhouse in Spain

Salmonellosis is a major foodborne infection in Spain, and strains that are resistant to a great variety of antibiotics have become a major public health concern. The aim of this study was to determine the level of antibiotic resistance in 133 Salmonella isolates obtained from a poultry slaughterhouse in Zaragoza (NE Spain). Antimicrobial resistance testing was performed by disk diffusion method using 19 antibiotics. Results were interpreted following the NCCLS criteria. Overall, the highest percentage of resistance was found to the following antimicrobial agents: sulfadiazine (96.2%), neomycin (53.4%), tetracycline (21.8%), and streptomycin (11.3%). All isolates were found to be resistant to one or more of the antibiotics tested. Multiple resistance was observed in 87 strains (65.4%). We found 23 different patterns of resistance in Salmonella Enteritidis. Resistance to sulfadiazine was the most common single resistance. The most frequent patterns of multiresistant strains were neomycin+sulfadiazine and neomycin+tetracycline+sulfadiazine. S. 4,5,12:b: showed the highest percentages of resistance to the tested drugs, with five different resistance patterns found. Ampicillin+chloramphenicol+streptomycin+sulphonamides+tetracycline (ACSSuT) resistance pattern, commonly associated with S. Typhimurium DT 104, was not detected in strains of the same phage type from broilers. The appearance of substantial multiresistance in foodborne Salmonella isolates suggests the need for more prudent use of antibiotics by farmers, veterinarians, and physicians.     

屠宰前鸡、猪源大肠杆菌耐药性调查

本试验旨在了解屠宰前鸡、猪源食品动物体内大肠杆菌耐药情况,分析潜在的食品安全问题。从广州市畜禽交易市场随机采集待屠宰鸡和猪的粪便样品,分离鉴定大肠杆菌,并采用琼脂稀释法检测大肠杆菌对15种抗菌药物的敏感性。结果显示,从658份猪源样品和133份鸡源样品中共分离鉴定出731株大肠杆菌,其中猪源606株,鸡源125株。药敏试验结果显示,731株大肠杆菌均表现出不同程度的耐药,耐药谱广且多重耐药现象严重。对复方新诺明和四环素的耐药率为90.0%以上,仅对头孢西丁、黏菌素和阿米卡星较敏感(耐药率均低于3%)。鸡源大肠杆菌对头孢噻肟、头孢曲松、新霉素、阿米卡星、萘啶酸和环丙沙星的耐药率显著高于猪源大肠杆菌(P＜0.05)。鸡源大肠杆菌中3耐及3耐以上的菌株占97.60%,猪源大肠杆菌占94.72%。结果表明,屠宰前畜禽体内大肠杆菌对临床常用抗菌药物的耐药性非常严重,以多重耐药为主,且耐药谱丰富多样。提示屠宰前畜禽携带的耐药菌对食品安全和人类健康存在较大的安全隐患。     

Reproduction of fowl typhoid by respiratory challenge with Salmonella Gallinarum

Fowl typhoid is a disease of adult chickens and is caused by Salmonella Gallinarum infection via the alimentary tract. The experimental reproduction of fowl typhoid per os (PO) requires artificial conditions to minimize the effect of gastric acid, and several Salmonella serovars have been known to be transmitted via the respiratory route. Therefore, we have hypothesized the existence of a respiratory route for Salmonella Gallinarum infection and have attempted to reproduce fowl typhoid via intratracheal challenge. In accordance with our hypothesis, the intratracheal challenges of Salmonella Gallinarum reproduced exactly same lesions as fowl typhoid and induced higher mortality and morbidity than those of the PO challenge. Therefore, this study represents the first reproduction of fowl typhoid via respiratory route, and our findings may be useful for understanding the transmission of Salmonella Gallinarum in the field.