首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 15 毫秒
1.
Three-hundred and forty-five herds (17 swine, 122 dairy sheep, 124 beef and 82 dairy cattle) were investigated for prevalence of Shiga toxin-producing Escherichia coli (STEC). Rectal faecal samples were selectively enriched and then examined by immunodetection techniques (Immunomagnetic Separation with anti-E. coli O157 Dynabeads, ImmunoMagnetic cell Separation (IMS) and automated enzyme-linked fluorescent immunoassay using VIDAS) and polymerase chain reaction (PCR) (rfbE and fliC genes) to assess the prevalence of E. coli O157:H7. Prevalence of non-O157 STEC was estimated by PCR screening for stx genes of 10 lactose-positive colonies grown on MacConkey agar after enrichment. PCR was used on all STEC isolates to detect stx(1), stx(2), eaeA and E-hlyA genes. Both immunodetection methods showed a moderate-good level of agreement (kappa = 0.649) but IMS showed 87.5% complementary sensitivity. Prevalence of positive herds for E. coli O157:H7 was estimated at 8.7% for sheep and 3.8% for cattle, whereas all the porcine herds tested negative. Non-O157 STEC were also absent from swine, but were isolated more frequently from ovine (50.8%) than bovine herds (35.9%). Within-herd prevalences of excretion of E. coli O157:H7 established by individual testing of 279 sheep (six herds) and 30 beef cattle (one herd) were 7.3% and 6.7% respectively. PCR analysis of 49 E. coli O157:H7 and 209 non-O157 isolates showed a different distribution of virulence genes. All E. coli O157:H7 were stx(2) gene-positive, eaeA was detected in 95.9%, and the toxigenic profile stx(2)/eaeA/E-hlyA was present in 75.5% of the isolates. Among the non-O157 STEC, prevalence of eaeA was significantly lower (5.3%) and E-hlyA was present in 50.2% of the isolates but only sporadically associated with eaeA. stx(2) was predominant in non-O157 isolates from cattle, whereas in sheep the combination stx(1)/stx(2) was more prevalent. This study demonstrated the wide distribution of STEC in ruminant herds, which represent an important reservoir for strains that pose a potential risk for human infections.  相似文献   

2.
3.
To estimate the prevalence of Escherichia coli O157 on Dutch dairy herds, faecal samples were collected once from 678 randomly selected dairy farms in the period October 1996-December 2000. Samples were cultured for E. coli O157. Thirty-eight isolates were tested for virulence genes (eae, VT1 and VT2). A questionnaire about farm characteristics was taken from the farm manager, resulting in variables that could be analysed to identify and quantify factors associated with presence of E. coli O157. In total, 49 of the 678 herds (7.2%) showed at least one positive pooled sample. E. coli O157 was not isolated from herds sampled in December-April in consecutive years (except for one isolate found in March, 2000). VT- and eae-genes were found in 37 and 38 isolates, respectively. Logistic regression was performed on variables obtained from the questionnaire, comparing E. coli O157-positive herds to negative herds. To account for season, a sine function was included in the logistic regression as an offset variable. In the final model, the presence of at least one pig at the farm (OR = 3.4), purchase of animals within the last 2 years before sampling (OR = 1.9), supply of maize (OR = 0.29) to the cows, and sampling a herd in the year 1999 or 2000 (compared to sampling in 1998; OR = 2.1 and 2.9, respectively) had associations with the presence of E. coli O157.  相似文献   

4.
5.
Shiga toxin-producing Escherichia coli (STEC) are an important group of emerging pathogens, with ruminants recognised as their main natural reservoir. The aim of this work was to establish the prevalence of non-O157 STEC in free-ranging wild ruminants in the Extremadura region of Spain and to characterise them phenogenotypically. Faecal samples were collected from 243 wild ruminants, including Cervus elaphus, Capreolus capreolus, Dama dama and Ovis musimon and were examined for STEC using both phenotypic (Vero cells) and genotypic (PCR and PFGE) methods.Shiga toxin-producing Escherichia coli were isolated from 58 (23.9%) of the samples and a total of 65 isolates were characterised. A PCR method indicated that 11 (16.9%) strains carried the stx1 gene, 44 (67.7%) carried the stx2 gene and 10 (15.4%) carried both these genes. The ehxA gene was detected in 37 (57%) of the isolates but none contained either the eae or saa genes. The isolates were from a total of 12 ‘O’ serogroups, although 80% were restricted to the O2, O8, O128, O146, O166 and O174 serogroups. The most commonly isolated STEC bacteria, which were from the O146 serogroup, exhibited a high degree of polymorphism as indicated by PFGE. Shiga toxin-producing Escherichia coli isolates of serogroups O20, O25, O166, O171, O174 and O176 had not previously been found in wild ruminants. This is the first study to confirm that wild ruminants in Spain are a reservoir of STEC and are thus a potential source of human infection.  相似文献   

6.
Four hundred and twenty-two calves were examined for intestinal carriage of Shiga toxin-producing Escherichia coli O157:H7 using conventional plating. Two (0.5%) E. coli O157 were recovered. They were compared with 96 Argentine strains of different origin by pulsed-field gel electrophoresis, phage typing and PCR-RFLP of stx2 genes. One strain isolated from a calf, was closely related with 18 strains of clinical origin.  相似文献   

7.
Little is known of the association of enterohemorrhagic Escherichia coli O157:H7/NM (EHEC O157) with disease in naturally infected calves, although cattle have been known as a major source for EHEC O157 outbreaks in humans. In this study, we investigated the occurrence of EHEC O157 in calves associated with/without diarrhoea to examine if EHEC O157 is involved in calf diarrhoea and to characterize the isolates. Four hundred and ninety eight diarrhoeic and non-diarrhoeic young calves from 115 different farms were examined. Of 244 diarrhoeic calves, 24 (9.8%) were positive for EHEC O157, and of 254 non-diarrhoeic calves, 7 (2.8%) were positive. EHEC O157 was recovered from 12/76 (15.79%) of diarrhoeic calves less than 2-week-old, and no EHEC O157 was detected in this age group of non-diarrhoeic calves. This implicates EHEC O157 as a possible cause of the disease in naturally infected neonatal calves. The occurrence of EHEC O157 was relatively lower in the older calves (aged older than 8 weeks) and no significant difference was found in the occurrence rates between these diarrhoeic and non-diarrhoeic calves. PCR analysis of virulence markers revealed that the isolates carried various virulence genes such as Ehly, eae, stx1 and stx2, which underlines the potential importance of these attributes for the infection, colonization and possible pathogenesis of calf diarrhoea.  相似文献   

8.
A risk-factor study was performed in eight dairy herds found to excrete verocytotoxin-producing Escherichia coli (VTEC) O157 in a former prevalence study. Associations between excretion of VTEC O157 and management factors such as housing and feeding were analysed in a generalised linear mixed model. The animals were stratified in three age groups and sampled four times during 1 year. The risk of excreting VTEC O157 was higher among weaned calves than non-weaned calves. Among the calves aged 1–4 months, the risk was reduced if the calf had suckled colostrum from the mother or if the calf had stayed >2 days with the mother after calving. Calves aged 5–24 months that had been moved within the last 2 weeks had a higher risk, but risk was reduced if fed barley silage. Cows fed grain or molasses had a higher risk of excreting VTEC O157.  相似文献   

9.
Verocytotoxin-producing Escherichia coli O157:H7 (VTEC O157:H7) was detected in two of 2446 individual faecal samples collected from pigs slaughtered at five Swedish slaughterhouses, indicating a prevalence of 0.08 per cent, with a 95 per cent confidence interval from 0 to 0.16 per cent Four Swedish VTEC O157:H7-positive farms which kept ruminants and pigs were studied by repeated faecal sampling; VTEC O157:H7 was isolated from the ruminants and pigs on all the farms and the same strains were present in the pigs and the ruminants. On one of the farms, the organism persisted in the pig population for 11 months. On all four farms, management practices which might have influenced the isolation rate in pigs were identified. A group of young VTEC O157:H7-positive pigs was moved from one of the VTEC O157:H7-positive farms to a fattening herd where there were no ruminants. The number of VTEC O157:H7-positive faecal samples decreased gradually and after nine weeks the pigs were all negative; at slaughter none of the pigs was VTEC O157:H7-positive.  相似文献   

10.
Faecal samples were collected from 573 slaughtered cattle aged between three and 24 months in seven abattoirs. After enrichment (mTSB with novobiocin), samples were screened by real‐time PCR first for stx and if positive, tested for the top‐five Shiga toxin‐producing Escherichia coli (STEC) serogroups using PCR assays targeting genes specific for serogroups O26, O103, O111, O145 and O157. Of 563 samples with available results, 74.1% tested positive for stx genes. Amongst them, the serogroups O145, O103, O26, O157 and O111 were detected in 41.9%, 25.9%, 23.9%, 7.8% and 0.8%, respectively. From 95 O26, 166 O145 and 30 O157 PCR‐positive samples, 17 O26, 28 O145 and 12 O157 strains were isolated by colony hybridization after immunomagnetic separation. The 17 O26 strains were eae‐positive, but only nine strains harboured stx (eight possessing stx1 and one stx2). Of the 28 O145 strains, ten were eae‐positive including four harbouring stx1 or stx2, whereas 18 were negative for stx and eae. Five of the 12 O157 strains harboured stx2 and eae, did not ferment sorbitol, and were identified as STEC O157:H7/H?. The other seven O157 strains were negative for stx and eae or positive only for eae. Shiga toxin genes and the top‐five STEC serogroups were frequently found in young Swiss cattle at slaughter, but success rates for strain isolation were low and only few strains showed a virulence pattern of human pathogenic STEC.  相似文献   

11.
利用生物信息学分析大肠杆菌O157:H7鞭毛蛋白FliC的二级结构及亲水性、抗原指数、柔性区域和表面可能性等指数,预测大肠杆菌O157:H7鞭毛蛋白FliC的潜在B细胞抗原表位,为其致病性研究提供理论基础。利用DNAStar软件Protean程序中Garnier-Robson方法和Chou-Fasman方法分析鞭毛蛋白FliC的α-螺旋、β-折叠、转角区域和卷曲区域,通过Kyte-Doolittle方法、Karplus-Schulz方法、Emini方法和Jameson-Wolf方法分析鞭毛蛋白FliC的亲水性、柔性区域、表面可能性和抗原指数。综合分析得出鞭毛蛋白FliC 63-74、236-247、338-349、460-471、542-553位氨基酸序列为潜在的B细胞优势抗原表位。化学合成法合成优势抗原表位338-349和460-471肽段,免疫BALB/c小鼠3次后,采用ELISA方法验证抗体水平。ELISA结果显示,338-349、460-471肽段具有很强的抗原性,能引起BALB/c小鼠产生高滴度的抗体。  相似文献   

12.
The prevalence of verocytotoxin-producing Escherichia coli (VTEC) O157 in 12-30-month-old beef finishing cattle in Scotland was determined using 1g faeces samples enriched in buffered peptone water, followed by immunomagnetic separation (IMS) and isolation on sorbitol MacConkey agar with cefixime and tellurite supplement (CT-SMAC). A validated questionnaire was used to collect information that could be associated with the samples. Generalised Linear Models and Generalised Linear Mixed Models were used to identify factors associated with shedding both between and within groups. A total of 14,856 samples were collected from 952 farms, of which 1231 were positive for VTEC O157. Prevalence levels were calculated with 95% confidence intervals as follows: 7.9% (6.5%, 9.6%) of animals sampled were estimated to be shedding VTEC O157, while 22.8% (19.6%, 26.3%) of farms were estimated as having at least one animal shedding in the group sampled. The median percentage of animals shedding in positive groups was 25% (20%, 32%). An increased probability of a group containing a shedding animal was associated with larger numbers of finishing cattle, the presence of pigs on the farm, or the farm being classed as a dairy unit stocking beef animals. Farms that spread slurry on grazing land were more likely to have shedding animals, while those that spread manure were at lower risk. Groups with older animals were less likely to be identified as positive. There was no significant regional difference in group shedding probabilities, but the proportion of positive groups dropped over two successive years of the study. Higher mean levels of shedding in positive groups were associated with animals being housed rather than at pasture, and this effect was stronger in groups which had recently had a change in housing or diet. Farms with animals at pasture had lower mean prevalence where water was supplied from a natural source, as had farms with higher numbers of finishing cattle. There remained unexplained variability in mean prevalence levels on positive farms in different areas of Scotland.  相似文献   

13.
A 1-year study on the animal-level prevalence and concentration of Escherichia coli O157 in adult sheep at slaughter was performed, to collect qualitative and quantitative information on the diffusion of the pathogen in adult sheep from Italy. A total 533 samples were collected, with a similar distribution in the four seasons. For prevalence estimates, a simple random sampling technique was used. An immuno-magnetic separation technique was used for sample screening, with enumeration of the pathogen in positive samples, along with molecular and serological identification of isolates. An overall prevalence of 7.1% (38/ 533, 95% CI 4.9-9.3%) was observed for fully virulent E. coli O157. A wide interval of VTEC O157 per gram was observed (< 100 to 6 x 10(5) CFU g(-1)), with 28.9% (11/38) of positive samples > or = 1 x 10(3) CFU g(-1), set as the threshold for those animals defined 'active shedders' for the purpose of the study. Eight per cent (3/38) of animals shed > 1 x 10(4) g(-1) VTEC O157, which represents > 96% of the total VTEC O157 bacteria cultured from all animals tested. The prevalence estimate of active shedders was therefore 2.1% (95% CI 0.9-3.3%). Most (34/38, 89.5%) of the positive animals were found in summer (July-September). Prevalence and concentrations of virulent VTEC O157 obtained in this study contribute to the demonstration that adult sheep represent a relevant source of environmental contamination from virulent VTEC O157, as well as a source of VTEC O157 contamination for food of ovine origin (meat and dairy products), especially during warm months.  相似文献   

14.
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.  相似文献   

15.
Equine piroplasmosis (EP) is caused by Theileria equi and/or Babesia caballi and has economic importance particularly in equines reared in poor management systems. This study is based on cELISA test to study the seroprevalence of EP among 370 horses and 150 donkeys in four Governorates north Egypt. Additionally, its risk factors were studied for the first time. The seroprevalence rates 36.5 %, 20 %, and 5.6 % for T. equi, B. caballi, and mixed infections, respectively. The highest antibody levels against T. equi were detected in Kafr ElSheikh (40 %) and Giza (40.1 %) Governorates, whereas those of B. caballi were detected in Qalyubia (25 %) and Kafr ElSheikh (24.1 %) Governorates. Concerning T. equi, animals >10 years (OR = 2.06) were more likely to be infected with EP than those <5 years old. In addition, the seropositivity increased among grazing (OR = 5.7, 95 % CI: 1.73−19.27) males (OR = 1.8, 95 % CI: 1.23−2.61) infested with ticks (OR = 2.3, 95 % CI: 1.60−3.48) during summer (OR = 4.3, 95 %CI: 2.53−7.46); whereas the seropositivity of animals for B. caballi increased among grazing equines (OR = 7.8, 95 % CI: 1.05−58.25) over 10 years old (OR = 2.08, 95 % CI: 1.10−3.94) and infested with ticks (OR = 2.4, 95 % CI: 1.54−3.76) during summer (OR = 7.12, 95 % CI: 3.15–16.06). Therefore, EP is an important prevalent disease in Egypt and deserves further attention regarding the management system, treatment, and vector control.  相似文献   

16.
In the period October 1996 through December 2000, a total of 7163 pooled faecal samples of laying hen and broiler flocks, finishing-pig herds and veal herds were examined for the presence of Salmonella spp., Campylobacter spp. and verocytotoxin-producing Escherichia coli O157 as part of a national monitoring programme in The Netherlands. Isolates were tested for eae and VT genes. Risk factors for Dutch veal herds were quantified. For all herd/flock types, faecal samples were cultured for E. coli O157. Of broiler flocks, laying flocks and finishing pig herds, respectively, 1.7%, 0.5% and 0.4% were E. coli O157 positive. In total, 42 of the 454 veal herds (9.3%) showed at least one positive pooled sample. E. coli O157-positive herds were compared (with logistic regression) to negative herds, regarding variables obtained from the questionnaire taken from the farm manager. To account for season, a sine function was included in the logistic regression as offset variable. In the final model, 'pink-veal production' (compared to white-veal production), 'group housing of the sampled herd' (compared to individual housing), 'more than one stable present' (compared to one stable present), 'hygienic measures regarding visitors' (compared to no hygienic measures), 'interval arrival-sampling of a herd of >20 weeks' (compared to < or =10 weeks), and 'presence of other farms within 1 km' (compared to no presence of farms <1 km) showed associations (P<0.05) with the presence of E. coli O157. These results need careful interpretation; they should be considered as indications for further (experimental or cohort-based) research rather then causal associations.  相似文献   

17.
为了了解大肠杆菌O157分离菌株携带毒力及黏附相关基因的情况及菌株的多态性.用聚合酶链式反应扩增stx1、stx2、eaeA、ehxA、EspA和Tccp基因.选用限制性内切酶HincⅡ和EcoRⅡ对分离的O157:H7菌株eaeA基因进行酶切,选用限制性内切酶HaeⅢ、HinfⅠ、EcoRⅡ和RsaⅠ对分离的O157:H7菌株chxA基因进行酶切,最后对这两个基因进行PCR-RFLP分析.结果,所有O157:H7菌株扩增出eaeA、ehxA、EspA和Tccp基因,但没有扩增出stx1和stx2基因;4株O157:H?菌株,均没有扩增出stx2、eaeA、ehxA、EspA和Tccp基因,且只有1株扩增出stx1基因.所有分离菌株的eaeA基因和ehxA基因选用限制性内切酶酶切之后所得酶切片段数目和大小与标准菌株的eaeA基因和ehxA基因选用限制性内切酶酶切之后所得酶切片段数目和大小相同.结果表明,由于所有菌株缺失stx2基因,其致病力相对较低,且在基因水平较为保守,多态性较为单一.  相似文献   

18.
Besides the well-known O157:H7 clone causing enterohaemorrhagic colitis and haemolytic uraemic syndrome in Europe, Japan and North America, the number of Escherichia coli isolates with non-motile (NM) phenotype has considerably increased. We supposed that spontaneous antibiotic resistance mutation could cause this phenotypic change. To model our hypothesis we isolated rifampicin--(Rif) and ampicillin--(Amp) resistant mutants from E. coli O157:H7 prototype strains 7785 and EDL933. Among Rifr mutants we could isolate strains with no or reduced motility, while the Ampr mutants became hypermotile. The biochemical profile of the mutants had not changed but phage sensitivity and generation time of the mutants were altered. Among the representative strains we did not find polymorphism with Southern blot analysis and no polymorphism was found in the fliC gene of the mutants. The described characteristics have proven to be stable. In a mice virulence assay by intravenous infections the virulence of the derivatives was also found to be changed. In summary, we found that the antibiotic-resistant phenotype in E. coli O157:H7 was coexpressed with several other phenotypic changes including motility and virulence. It can be assumed that expression of the involved phenotypes may be under the influence of a common regulatory cascade. Further work is needed to identify the components and mechanism of this regulatory system.  相似文献   

19.
Escherichia coli isolates from calves were investigated by multiplex PCR assays for the presence of genes encoding K99, F41, F17-related fimbriae, heat-stabile enterotoxin a (STa), intimin (eae) and Shiga toxins (stx1 and stx2). A total of 120 E. coli isolates, 75 isolated from diarrhoeic or septicemic calves and 45 from clinically healthy calves aged between 1 day and 2 months were tested. Each isolate was obtained from different calves in different herds. Among the isolates from diseased animals, 12 (16%) isolates from 1- to 7-day-old diarrhoeic calves were detected as enterotoxigenic E. coli which possessed K99, F41 and STa in combination; F17-related fimbriae genes were detected in 33 (44%) isolates and they were found in combination with K99 + F41 + STa in two isolates. Of 120 isolates, 16 carried eae, eight stx1 and five stx2 genes alone or in combination. None of the eae- or stx-positive strains was identified as O157:H7. However, results indicate that calves may be carrier of Shiga toxin-producing E. coli which have potential as a human pathogen. Antimicrobial susceptibility of 75 isolates from diseased calves was determined by agar disk diffusion method for 14 antimicrobial agents. In 77.3% of the isolates, multiresistance was detected. Higher resistance rates were detected for cephalothin (72%), tetracycline (69.3%), kanamycin (69.3%), ampicillin (65.3%), nalidixic acid (53.3%), trimethoprim-sulphamethoxazole (52%) and enrofloxacin (41.3%), respectively. No resistance was found for ceftiofur and cefoxitin.  相似文献   

20.
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.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号