Factors associated with the presence of Escherichia coli O157 in feedlot-cattle water and feed in the Midwestern USA

Our objective was to generate hypotheses about associations between management, climate, and the presence of Escherichia coli O157 in feedlot–cattle water tanks and in feedlot–cattle feed. Water samples from 710 tanks on 73 feedlots, and feed-samples from a subset of 504 pens on 54 feedlots, in four US states were tested for E. coli O157. Management and climate factors were ascertained by survey and observation. Escherichia coli O157 were isolated from 13% of the water tanks and at least one water tank was positive on 60% of the feedlots. The factors significantly associated with E. coli O157 in water were greater percentage of cattle shedding E. coli O157 in faeces within the same pen, higher concentration of total E. coli in the water, lack of the clarity of the water, the use of fly traps, the reported frequency of rodent sightings in the pen or alley area, and the weather at the time of sampling. Escherichia coli O157 were isolated from 14.9% of the feed samples obtained from the feedbunks. Factors positively associated with E. coli O157 in feed were higher heat index at the time of sampling, the presence of cottonseed meal in the ration, and the feedlot location (state). Coliform counts in feed, presence of E. coli O157 in water tanks and faecal prevalence of E. coli O157 were not associated with the presence of E. coli O157 in feed.     

新疆牛源大肠杆菌O157∶H7的分离鉴定及ERIC-PCR基因分型研究

为了了解牛源大肠杆菌（E.coli）O157∶H7在新疆地区的污染状况以及遗传多样性,探究不同地区分离菌株的遗传关系,为控制牛源E.coli O157∶H7的传播提供试验依据。将采集的样品在EC肉汤中进行增菌（37 ℃、180 r/min）,接着将增菌液划线接种到SMAC平板上,37 ℃培养箱中过夜培养18 h左右。挑取SMAC平板上白色或无色单菌落接种MUG培养基,37 ℃培养18 h左右,将无荧光样品接种到SMAC平板上,37 ℃培养18 h左右,隔天挑取白色或无色单菌落进行PCR鉴定,具有rfbE和fliC基因条带的即为阳性菌株。将阳性菌株进行肠杆菌基因间重复共有序列扩增（ERIC-PCR）指纹图谱聚类分析,分析菌株之间的同源性关系。ERIC-PCR结果显示,相似性100%的菌株有3组。从伊犁地区分离到的菌株差异性最大,具有6种分型;其次是乌鲁木齐,具有4种分型。菌株来源多样性最多的在D簇,由此可见通过ERIC-PCR分型,可以进行溯源观察。ERIC-PCR能够区分特定采样点或物种的分离物,它能够证明从不同来源的菌株之间,存在着某些相似的ERIC特性,并聚集在同一个簇群中。该研究中筛选出的E.coli O157∶H7菌株具有广泛的遗传多样性,该方法对于检测不同物种间的细菌差异非常敏感。由此可见ERIC-PCR可以作为E.coli O157∶H7常规监测和鉴定的一个有效的工具。     

猪源大肠杆菌O157:H7体外氟苯尼考耐药诱导及对抗菌药物敏感性变化研究

为初步研究猪源大肠杆菌O157:H7 (E.coli O157:H7)对氟苯尼考耐药性的产生和消除机制,本研究采用亚抑菌浓度体外耐药诱导的方法将两株猪源大肠杆菌O157:H7诱导成氟苯尼考高度耐药菌株,采用无氟苯尼考压力下连续传代培养的方法将获得的氟苯尼考耐药菌株的氟苯尼考耐药性消除,检测耐药诱导菌和耐药消除菌对抗菌药物的敏感性,并检测菌株质粒携带的耐药基因。结果显示,经氟苯尼考耐药诱导,猪源大肠杆菌O157:H7对氟苯尼考、阿莫西林、头孢唑啉、头孢拉定和头孢噻吩由敏感变为耐药,对头孢噻肟的敏感性由敏感变为中介,对氧氟沙星、环丙沙星和阿奇霉素由中介变为耐药;而经耐药消除后,菌株恢复对上述药物的敏感性;在菌株的质粒中检测到氟苯尼考耐药基因、喹诺酮类耐药基因和β-内酰胺酶基因,与耐药表型相符。结果表明,在氟苯尼考压力的长期存在下,猪源大肠杆菌O157:H7对氟苯尼考产生耐药,且对青霉素类、头孢类和喹诺酮类药物产生交叉耐药,在去除氟苯尼考压力下连续培养,可消除菌株的部分耐药性。     

牛源大肠杆菌对秀丽隐杆线虫的致病性研究

为建立牛源大肠杆菌-秀丽隐杆线虫致病模型，本研究从广西南宁、桂林、柳州等地区收集的牛病料中分离出13株大肠杆菌，继而对这些大肠杆菌进行血清学鉴定及小鼠、秀丽隐杆线虫的致病性试验。采用玻片凝集法鉴定大肠杆菌的O血清型，并将13株大肠杆菌培养液以3.0×10⁹ CFU/mL、0.2 mL/10 g体重给小鼠腹腔注射，同时分别喂食N2野生型秀丽隐杆线虫。结果显示，大肠杆菌的O血清型为O127、O126和O44，其中O126为优势血清型（4/13）。致病性结果显示，有11株大肠杆菌能使小鼠致死（致死率为40%~100%），2株大肠杆菌对小鼠没有致死性（致死率为0）。对小鼠有强致病性的大肠杆菌，对线虫的致死率也较高，死亡率在第3~6天最为显著，半数致死时间为3~4.5 d，最长存活时间为9 d；对小鼠不致死的两株大肠杆菌对线虫的致死率也较低，线虫死亡率下降趋势缓慢，半数致死时间为5~6 d，最长存活时间为10~11 d。肠道细菌计数结果显示，大肠杆菌在线虫体内的数量与时间呈线性关系，大肠杆菌不断破坏线虫的免疫系统，从而导致了线虫的死亡。本研究结果表明，大肠杆菌对线虫和小鼠的致病力试验结果一致，说明成功建立了牛源大肠杆菌-秀丽隐杆线虫致病模型，为牛病防治与临床用药提供了依据。     

Risk factors for hide contamination of Scottish cattle at slaughter with Escherichia coli O157

In the slaughter processing of cattle, contaminated hides have been identified as one of the major sources of Escherichia coli O157 carcase contamination. Logistic regression analysis was applied to data collected in a large scale study in Scotland involving 222 cattle forming 34 groups sent for slaughter from 30 farms to 10 slaughterhouses. Aspects of individual animal characteristics, farm management practices and slaughterhouse features were examined to identify potential risk factors for hide contamination at harvest. Two models were developed, the first in which slaughterhouse was modelled as a fixed effect, and a second model where slaughterhouse and farm groups were modelled as random effects. In the first model, there was a significantly increased risk of a carcase testing positive for E. coli O157 on the hide if either the hide of the carcase immediately before or after it on the line was contaminated (OR 3.6; 95% CI: 1.4–9.9). If both adjacent carcases had contaminated hides, the odds ratio for the study carcase having a contaminated hide rose to 11.5 (95% CI: 4.4–32.5). If animals were held in lairage, receiving hay as feed appeared to have a protective effect on hide contamination. Transportation to the slaughterhouse by haulier, as opposed to transport by the farmer, was associated with a 5.4 increase in the odds of E. coli O157 contamination. The use of a crush in the lairage, often employed when reading ear tags, was also found to significantly increase the odds of hide contamination with E. coli O157. In the second model, the inclusion of slaughterhouse and farm group as random effects resulted in two of the previously identified factors being associated with hide contamination. If at least one of the adjacent carcases on the line had a contaminated hide, the associated odds ratio was 6.6 (95% CI: 2.8–15.9), which rose to 22.7 (95% CI: 9.3–55.5) if both adjacent hides were contaminated. Receiving hay in lairage was found to be important to the model, although not significant in itself (OR 0.005; 95% CI: 1.2e⁻⁶–20.7). These results suggest that modifiable risk factors for hide contamination exist. However, in order best to reduce the prevalence of hide contamination at slaughter, individual slaughterhouse risk assessment and intervention strategies are appropriate.     

Escherichia coli O157:H7 vaccine field trial in 9 feedlots in Alberta and Saskatchewan

A feedlot trial was conducted to assess the efficacy of an Escherichia coli O157:H7 vaccine in reducing fecal shedding of E. coli O157:H7 in 218 pens of feedlot cattle in 9 feedlots in Alberta and Saskatchewan. Pens of cattle were vaccinated once at arrival processing and again at reimplanting with either the E. coli O157:H7 vaccine or a placebo. The E. coli O157:H7 vaccine included 50 microg of type III secreted proteins. Fecal samples were collected from 30 fresh manure patties within each feedlot pen at arrival processing, revaccination at reimplanting, and within 2 wk of slaughter. The mean pen prevalence of E. coli O157:H7 in feces was 5.0%; ranging in pens from 0% to 90%, and varying significantly (P < 0.001) among feedlots. There was no significant association (P > 0.20) between vaccination and pen prevalence of fecal E. coli O157:H7 following initial vaccination, at reimplanting, or prior to slaughter.     

Management and nutritional factors associated with the detection of Salmonella sp. from cattle fecal specimens from feedlot operations in the United States

In a convenience sample of 100 feedlot operations (included in the United States Department of Agriculture: Animal and Plant Health Inspection Service 1994 Cattle on Feed Evaluation), up to 25 cattle fecal samples were collected and tested for the presence of Salmonella from each of two pens (the pen which contained the most-recent arrivals, and the pen with cattle that had been on feed the longest). One or more Salmonella spp. were recovered from 38 (38.0%) of the 100 feedlots, 52 (26.0%) of the 200 pens and 273 (5.5%) of the 4977 fecal samples collected. Multivariable logistic regression indicated that feeding tallow and feeding whole cottonseed or cottonseed hulls within seven days prior to fecal sample collection was associated with an increased risk of finding Salmonella in a pen. Variables not found to be significantly associated with the detection of Salmonella in a pen included region, operation size, use of sprinklers, time on feed, type of cattle in the pen, number and concentration of cattle in a pen, feeding probiotics, and various other feeds.     

微量凝集试验用大肠杆菌O50、O60、O117、O131定型血清的研制

为进一步完善中国大肠杆菌菌体微量凝集试验定型血清库，改良微量凝集试验用大肠杆菌O抗原定型血清的制备工艺，本试验选取大肠杆菌O50、O60、O117、O131血清型进行定型血清制备方法的探究。首先用大肠杆菌不同血清型的参考菌株制备灭活抗原，而后多次免疫家兔以采血获得粗血清，用微量凝集试验的方法确定不同粗血清的交叉凝集素及凝集效价，再通过交叉凝集素吸收法结合血清稀释法消除非特异性凝集，最终研制出特异性良好的微量凝集试验用大肠杆菌O抗原定型血清。本研究确定了大肠杆菌O50、O60、O117、O131定型粗血清的主要交叉凝集素，最终成功研制出特异性良好的微量凝集试验用大肠杆菌O50、O60、O117、O131定型血清共4种，凝集效价在1：256至1：2 048之间，其中微量凝集试验用大肠杆菌O50、O60定型血清无交叉凝集素，为单因子定型血清，微量凝集试验用大肠杆菌O117、O131定型血清存在1~2种非特异性的交叉凝集素O14和O107。本研究作为大肠杆菌O抗原定型血清制备工艺摸索过程中的重要部分，为完善中国大肠杆菌菌体微量凝集试验定型血清库，进一步改良微量凝集试验用大肠杆菌O抗原定型血清的制备方法提供了重要理论依据。     

鸭致病性大肠杆菌分离鉴定及其相关致病性分析

为研究鸭大肠杆菌的致病性及相关生物特性,本试验从西昌市某规模化鸭场采集病料,通过传统分离培养及理化性质鉴定,分离得到27株鸭致病性大肠杆菌。对27株鸭致病性大肠杆菌进行血清型鉴定、药敏试验、相关毒力基因检测。血清型鉴定结果显示,优势血清型为O119、O86、O126、O142和O44,占分离株55.56%。血清型O119占27株鸭致病性大肠杆菌的40.74%,为该鸭场流行的致病血清型。对20种兽医临床常用药物的药敏试验结果显示,27株鸭致病性大肠杆菌均对阿米卡星、庆大霉素和多黏菌素B敏感,对头孢曲松等10种药物较敏感,对利福平等5种药物耐受。大肠杆菌相关毒力基因检测结果显示, iutA、hlyF、Iss、IroN、ompT、fyuA、irp 2、Tsh及papA基因携带率均为100.00%, fimC基因携带率59.26%, K 99基因携带率7.40%。各项研究结果为有效防控鸭大肠杆菌病提供了重要科学依据,并为大肠杆菌深入研究奠定了基础。     

Dietary monensin level, supplemental urea, and ractopamine on fecal shedding of Escherichia coli O157:H7 in feedlot cattle

Inclusion of distillers grains (DG) in cattle diets has been shown to increase fecal shedding of Escherichia coli O157:H7. It is hypothesized that altered gut fermentation by DG may be responsible for the positive association. Therefore, feed additives affecting ruminal or hindgut fermentation of DG also may affect fecal shedding of E. coli O157:H7. The objectives of the study were to evaluate effects of monensin (33 or 44 mg/kg of DM), supplemental urea (0, 0.35, or 0.70% of DM), and ractopamine (0 or 200 mg/steer daily administered during the last 42 d of finishing) in a steam-flaked corn grain-based diet containing 30% wet sorghum DG on fecal shedding of E. coli O157:H7. Seven hundred twenty crossbred beef steers, housed in 48 pens (15 steers/pen), were assigned to dietary treatments in a randomized complete block design with a 2 × 3 × 2 factorial treatment arrangement. Fresh pen floor fecal samples (10 per/pen) were collected every 2 wk for 14 wk (July through November) and cultured for E. coli O157:H7. Isolation of E. coli O157:H7 was by selective enrichment of fecal samples in an enrichment broth, immunomagnetic separation, followed by plating onto a selective medium. Samples that yielded sorbitol-negative colonies, which were positive for indole production, O157 antigen agglutination, and contained rfbE, fliC, and stx2 were considered positive for E. coli O157:H7. Fecal prevalence data were analyzed as repeated measures using negative binomial regression to examine effects and interactions of sampling day, urea, monensin, and ractopamine. Mean fecal prevalence of E. coli O157:H7 was 7.6% and ranged from 1.6 to 23.6%. Cattle fed monensin at 44 mg/kg of feed had less (P = 0.05) fecal E. coli O157:H7 prevalence than cattle fed 33 mg/kg (4.3 vs. 6.8%). Although the reason for the reduction is not known, it is likely because of changes in the microbial ecosystem induced by the greater amount of monensin in the hindgut. Supplemental urea at 0.35 or 0.70% had no effect (P = 0.87) on fecal shedding of E. coli O157:H7. Fecal prevalence of E. coli O157:H7 were 5.3, 5.7, and 5.9% for groups fed 0, 0.35, and 0.7% urea, respectively. The inclusion of ractopamine at 0 or 200 mg/(animal?d) had no effect (P = 0.89) on fecal prevalence of E. coli O157:H7 (4.4 vs. 4.0%). Additional research is needed to confirm the reduction in fecal shedding of E. coli O157:H7 in cattle fed monensin at 44 mg/kg of feed compared with cattle fed 33 mg/kg of feed.     

Isolation,Identification and Related Pathogenicity Analysis of Duck Pathogenic E.coli

To analyze the pathogenicity and biological characteristics of duck E.coli, 27 strains of duck pathogenic E.coli were isolated from dead ducks with the typical characterize of colibacillosis by isolation and culture, physical and chemical properties identification in Xichang city.O serotype identification, drug sensitivity test and virulence associated genes were detected in 27 strains of duck pathogenic E.coli.The result showed that the predominant serotypes were O119, O86, O126, O142 and O44, which accounted for 55.56%.O119 was the epidemic and pathogenic serotype in this farm, which accounted for 40.74% in 27 strains of duck pathogenic E.coli.Through the drug sensitivity test of 20 kinds of clinical commonly used drugs, we found that all strains were sensitive to amikacin, gentamicin and polymyxin B.Besides, 10 drugs(ceftriaxone, etc) were lightly sensitive.However, these strains were resistant to 5 kinds of drugs(rifampicin, etc).The detection result of PCR about virulence associated genes indicated that the positive of iutA, hlyF, Iss, IroN, ompT, fyuA, irp 2, Tsh and papA genes were 100.00%, fimC and K 99 genes were 59.26% and 7.40%, respectively.The results provided an important reference for effective prevention and control of duck colibacillosis, and laid the foundation for further study of E.coli.     

新疆地区腹泻仔猪源大肠杆菌的分群、血清型鉴定及耐药性分析

旨在了解新疆地区腹泻仔猪源大肠杆菌的系统进化分群、血清型及耐药性。本研究对154份腹泻仔猪粪便样品进行大肠杆菌的分离鉴定,采用多重PCR方法对分离株进行系统进化分群和O血清型鉴定,通过K-B纸片法对其进行药物敏感性检测并通过PCR方法进行耐药基因检测。结果显示：共分离到154株大肠杆菌,包括ETEC(n=24)、STEC(n=21)、EPEC(n=1)、EPEC/STEC(n=2)、ETEC/STEC(n=1)和ETEC/EPEC(n=1),其他104株。系统进化分群显示,多数菌株属于B1(37%)和A群(31%)。定型菌株44株,分别属于10种血清型,以O154、O12、O8、O141和O175为主要流行血清型。151株(98%)为多重耐药菌,对复方新诺明、四环素、氨苄西林、链霉素和氯霉素的耐药率为81%～100%,对阿莫西林/克拉维酸、头孢噻肟、庆大霉素、头孢曲松、环丙沙星和阿米卡星的耐药率为31%～66%,对左氧氟沙星、多黏菌素B、头孢他啶、头孢吡肟、氨苄西林-舒巴坦、哌拉西林-他唑巴坦和亚胺培南的耐药率为1%～19%。耐药基因tetA(88%)、tetG(60%)和cmlA(4...     

东南沿海地区水禽源大肠杆菌耐药表型及耐药基因型的调查

为研究水禽中大肠杆菌的耐药表型及耐药基因型,本研究从中国广东、福建、浙江、江苏和山东5个省市的水禽养殖场及周边环境中采集鸭粪便样品、水样以及土样1 505份,采用选择性培养基和基质辅助激光解吸电离飞行时间(MALDI-TOF-MS)质谱分离鉴定大肠杆菌,采用微量肉汤稀释法检测耐药表型,采用PCR方法筛选大肠杆菌中的25种耐药基因。结果显示,1 505份样品中共分离鉴定出449株大肠杆菌,分离率为29.8%,其中335份来源于粪便,52份来源于土壤,62份来源于水,这些样品中浙江(43.3%)和广东(43.2%)的分离率最高,其次是福建(28.1%)和江苏(23.8%),山东(18.3%)的分离率最低。449株大肠杆菌具有多重耐药性,对氨苄西林、环丙沙星、金霉素、氟苯尼考和磺胺类药物的耐药率分别为90.2%、50.3%、96.7%、87.3%和90.6%,而对替加环素和美罗培南则比较敏感,来源不同省份的大肠杆菌对不同抗菌药呈现出不同的耐药率。PCR检测结果发现,449株大肠杆菌中耐药基因tet(A)检出率最高(88.2%),随后是floR(58.13%)、sul2(57.02%)、tet(B)(53.9%)、cmlA(39.2%)和sul1(36.75%),未检测到qnrA、tet(M)和tet(X)耐药基因的大肠杆菌。在这5个省份中,上述耐药基因在山东省样品中分布均较高,而在浙江省和江苏省样品中分布较低。综上所述,本研究调查了中国东南沿海五省水禽及环境中大肠杆菌的耐药现状,为养殖业抗生素的规范使用提供理论依据。     

3种食源性致病菌TaqMan多重荧光定量PCR检测方法的建立

旨在建立一种可同时快速检测大肠杆菌O157 ∶ H7、沙门菌和产单核细胞李氏杆菌3种食源性致病菌的TaqMan多重荧光定量PCR(qPCR)方法.针对大肠杆菌O157 ∶ H7 rfbE基因、沙门菌invA基因和产单核细胞李氏杆菌hlyA基因的保守序列分别设计特异性引物和TaqMan探针,建立多重qPCR反应体系,进行...     

Stochastic Simulation Model Comparing Distributions of STEC O157 Faecal Shedding Prevalence Between Cattle Vaccinated With Type III Secreted Protein Vaccines and Non‐Vaccinated Cattle

Pens of cattle with high Escherichia coli O157:H7 (STEC O157) prevalence at harvest may present a greater risk to food safety than pens of lower prevalence. Vaccination of live cattle against STEC O157 has been proposed as an approach to reduce STEC O157 prevalence in live cattle. Our objective was to create a stochastic simulation model to evaluate the effectiveness of pre‐harvest interventions. We used the model to compare STEC O157 prevalence distributions for summer‐ and winter‐fed cattle to summer‐fed cattle immunized with a type III secreted protein (TTSP) vaccine. Model inputs were an estimate of vaccine efficacy, observed frequency distributions for number of animals within a pen, and pen‐level faecal shedding prevalence for summer and winter. Uncertainty about vaccine efficacy was simulated using a log‐normal distribution (mean = 58%, SE = 0.14). Model outputs were distributions of STEC O157 faecal pen prevalence of summer‐fed cattle unvaccinated and vaccinated, and winter‐fed cattle unvaccinated. The simulation was performed 5000 times. Summer faecal prevalence ranged from 0% to 80% (average = 30%). Thirty‐six per cent of summer‐fed pens had STEC O157 prevalence >40%. Winter faecal prevalence ranged from 0% to 60% (average = 10%). Seven per cent of winter‐fed pens had STEC O157 prevalence >40%. Faecal prevalence for summer‐fed pens vaccinated with a 58% efficacious vaccine product ranged from 0% to 52% (average = 13%). Less than one per cent of vaccinated pens had STEC O157 prevalence >40%. In this simulation, vaccination mitigated the risk of STEC O157 faecal shedding to levels comparable to winter, with the major effects being reduced average shedding prevalence, reduced variability in prevalence distribution, and a reduction in the occurrence of the highest prevalence pens. Food safety decision‐makers may find this modelling approach useful for evaluating the value of pre‐harvest interventions.     

西藏那曲市羊源大肠杆菌分离鉴定及耐药性研究

为了解西藏那曲市羊大肠杆菌的耐药情况，指导临床进行合理用药，本试验从那曲市采集羊新鲜无污染腹泻物92份，进行大肠杆菌显色培养基分离、革兰氏染色镜检、生化鉴定、分子生物学鉴定、致泻性大肠杆菌生化鉴定、药敏试验及耐药基因检测。结果显示，分离菌株在大肠杆菌显色培养基上呈蓝色菌落、革兰氏染色为粉红色的短杆菌，通过生化鉴定及23S rRNA的PCR检测得到26株羊源大肠杆菌，分离率为28.3%；其中25株符合致泻性大肠杆菌生化特性，致泻菌株分离率为27.2%。药敏试验结果显示，所得25株羊源大肠杆菌对氨苄西林的耐药性较高，耐药率为24.0%；对羧苄西林、卡那霉素的耐药性次之，耐药率为8%；对哌拉西林、头孢呋辛、庆大霉素、四环素、米诺霉素等药物耐药率为4%；对诺氟沙星、氧氟沙星、环丙沙星等药物极为敏感，可作为临床用药。5种耐药基因检测结果显示，bla_TEM基因检出率为100%，表明分离菌均含有相应的耐药基因。以上结果表明，西藏那曲市羊源大肠杆菌对多种药物耐药，提示在临床实践过程中应注重合理用药、联合用药，减缓大肠杆菌耐药性的产生。     

Prevalence of intestinal pathogens in Danish finishing pig herds

Our aim was to determine the prevalence of the intestinal bacteria: Lawsonia intracellularis, Brachyspira hyodysenteriae, Serpulina intermedia, Brachyspira innocens, Brachyspira pilosicoli, pathogenic Escherichia coli (serogroups O138, O139, O141 and O149) and Salmonella enterica in Danish finishing pig herds. A total of 79 herds was randomly selected and visited during 1998. From each herd, 20 faecal samples were collected from individual pigs weighing 30–50 kg. Furthermore, 10 pooled pen samples were collected and examined for S. enterica. In total, 1580 faecal samples and 790 pen samples were collected and examined by polymerase chain reaction (PCR) or culture. L. intracellularis was found in 74 herds (93.7%), B. hyodysenteriae in two herds (2.5%), S. intermedia in 10 herds (12.7%), B. innocens in 27 herds (34.2%), B. pilosicoli in 15 herds (19.0%), pathogenic E. coli in 19 herds (24.1%) and S. enterica in eight herds (10.1%). The within-herd prevalences of L. intracellularis and B. hyodysenteriae were 25–30%; the within-herd prevalences of the other agents were 5–10%. Three herds (4%) were not infected with any of the bacteria and 25 herds (32%) were only infected with L. intracellularis.     

Characterisation of avian pathogenic Escherichia coli (APEC) associated with colisepticaemia compared to faecal isolates from healthy birds

A total of 114 avian pathogenic Escherichia coli (APEC) isolates were collected from cases of colisepticaemia occurring in broilers (77) and layers (37) within Ireland. In addition 45 strains isolated from faeces of healthy birds were included for comparison. All isolates were serogrouped, and examined for known virulence factors, mostly by PCR. The O78 serogroup represented 55 and 27% of broiler and layer colisepticaemic isolates respectively. All isolates were positive for curli fimbriae (crl, csg) and negative for afimbrial adhesin (afa). S-fimbrial (sfa) sequences were present in 8.8% of septicaemic isolates and 8.9% of healthy bird isolates. The majority of E. coli from cases of colisepticaemia (97.4%) and healthy bird (95.6%) isolates were positive for aerobactin (aer), and temperature sensitive haemagglutinin (tsh) was similarly detected in high numbers in 93.9 and 93.3%, respectively. In comparison to E. coli isolates from the faeces of healthy birds, a significantly higher percentage of isolates from septicaemic cases possessed Type 1 fimbriae (fimC) and increased serum survival (iss) gene sequences. Forty-seven (41.2%) isolates from septicaemic birds possessed P-fimbriae (pap) gene sequences, compared with only 15.6% from E. coli isolated from healthy birds. Haemolysin (hlyE) sequences were detected in 46.7% of isolates from healthy birds in comparison with 6.1% of septicaemic isolates. Sequences encoding colicin V (cvaC) were detected in 99.1% of septicaemic isolates and 82.2% of isolates from healthy birds. The K1 capsule was only present in two septicaemic isolates, both taken from layers. Motility was detected in 36.8% of E. coli isolated from cases of septicaemia, compared with 93.3% of isolates from healthy birds. These results demonstrate the presence of 11 virulence genes in E. coli isolated from cases of colisepticaemia within Ireland, and indicate the prevalence of iss and fimC.     

鸭源致病性大肠杆菌O抗原与毒力基因检测及药敏试验

【目的】探究荣昌、大足和隆昌三地鸭大肠杆菌分离株的O抗原、毒力基因及耐药性。【方法】将2014年—2021年鸭病料中分离得到的107株细菌在无菌条件下接种于麦康凯培养基中划线进行培养纯化,通过16S rDNA基因扩增测序和生化试验进行细菌鉴定,采用PCR技术对O抗原和16种毒力基因进行检测,采用Kirby-Bauer纸片扩散法进行药敏试验。【结果】107株分离株鉴定为大肠杆菌;O抗原鉴定试验鉴定出9种O抗原,其中优势抗原为O78(37.00%)、O7(25.00%),O121和O145(均为15.00%),并检测到5株O78+O145和O7+O145融合株;共检测出11种毒力因子,其中强致病性毒力基因有Tsh基因(检出率为25.23%)、fyuA基因(检出率为31.78%)、estB基因(检出率为31.78%)、Vat基因(检出率为2.80%)、iucA基因(检出率为44.56%)。3种毒力基因ompA、yijP和ibeB的携带率最高,分别为100.00%、96.26%和85.98%;药敏试验结果表明分离株对氨基糖苷类药物、米诺环素和多黏菌素最为敏感,对大环内酯类药物和克林霉素耐药,分...     

畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌的LAMP检测及其耐药性分析

为深入了解畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌的流行及耐药状况,采用建立的畜禽粪便中金黄色葡萄球菌和大肠埃希氏菌的环介导等温扩增技术（LAMP）检测方法对山东省内牛粪污、猪粪、鸡粪、鸭粪中的金黄色葡萄球菌和大肠埃希氏菌进行检测,进而对检测阳性样品进行分离,并对药敏试验中多重耐药性菌株进行耐药基因预测。结果表明,LAMP方法检测畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌与国家标准方法检测结果符合率为100%,重复性好,特异性高。从80份畜禽粪污样品中分离到的3株金黄色葡萄球菌均对青霉素耐药;34株大肠埃希氏菌对氟苯尼考、利福平的耐药比例高于50%,高于25%以上的还有氨苄西林、四环素、多西环素、磺胺异噁唑、头孢噻吩和头孢噻呋;耐药基因预测结果显示,鸡粪、鸭粪、牛粪和猪粪中的4株大肠埃希氏菌分别携带10、8、20和15种耐药基因;鸡粪中的1株金黄色葡萄球菌携带11种耐药基因。说明山东地区畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌耐药状况严峻,菌株普遍多重耐药,且携带多种耐药基因。