Prevalence and risk factors for Salmonella spp. and Campylobacter spp. caecal colonization in broiler chicken and turkey flocks slaughtered in Quebec, Canada

We conducted an observational study to estimate prevalence and risk factors for Salmonella spp. and Campylobacter spp. caecal colonization in poultry. Eighty-one broiler chicken and 59 turkey flocks selected among flocks slaughtered in the province of Quebec, Canada, were included in the study. Flock status was evaluated by culturing pooled caecal contents from about 30 birds per flock. Exposure to potential risk factors was evaluated with a questionnaire. Odds ratios were computed using multivariable logistic regression.

The prevalence of Salmonella-positive flocks was 50% (95% CI: 37, 64) for chickens and 54% (95% CI: 39, 70) for turkeys, respectively. Odds of Salmonella colonization were 2.6 times greater for chicken flocks which failed to lock the chicken house permanently. In turkeys, odds of Salmonella colonization were 4.8–7.7 times greater for flocks which failed to be raised by ≤2 producers with no other visitors allowed onto the premises, or origin from a hatchery.

The prevalence of Campylobacter-positive flocks was 35% (95% CI: 22, 49) for chickens and 46% (95% CI: 30, 62) for turkeys. Odds of colonization were 4.1 times higher for chicken flocks raised on farms with professional rodent control and 5.2 times higher for flocks with manure heap >200 m from the poultry house, and also increased with the number of birds raised per year on the farm and with the age at slaughter. For turkeys, odds of Campylobacter flock colonization were 3.2 times greater in flocks having a manure heap at ≤200 m from poultry house and 4.2 times greater in flocks drinking unchlorinated water.     

Prevalence and antimicrobial resistance among Escherichia coli and Salmonella in Ontario smallholder chicken flocks

Surveillance is an important component of an overall strategy to address antimicrobial resistant bacteria in food animals and the food chain. The poultry market has many points of entry into the Canadian food chain, and some production practices are underrepresented in terms of surveillance. For example, pathogen carriage and antimicrobial resistance surveillance data are limited in smallholder chicken flocks raised for slaughter at provincially inspected abattoirs. In Canada, antimicrobial resistance in Escherichia coli and Salmonella isolated from commercial broiler chicken flocks, slaughtered at federally inspected abattoirs, is monitored by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS ). The objective of this study was to establish baseline information of antimicrobial resistance presence in E. coli and Salmonella isolated from smallholder flocks in Ontario, utilizing CIPARS collection and isolation methodologies, and to compare findings with CIPARS federally inspected abattoir data from Ontario, Canada. Five chickens per flock were sampled from 205 smallholder flocks. Of 1,025 samples, the E. coli prevalence was 99% (1,022/1,025), and 47% (483/1,022) of positive E. coli isolates were resistant to one or more of the 14 antimicrobials. Furthermore, as compared to results reported for the CIPARS commercial flocks, E. coli isolates from smallholder flocks had significantly lower resistance prevalence to six of 14 individual antimicrobials. Recovery of E. coli did not differ between federally inspected and provincially inspected flocks. Salmonella prevalence at the bird level in smallholder flocks was 0.3% (3/1,025), significantly lower (p  ? 0.0001, 95% CI 0.080%–0.86%) than federally inspected commercial flocks. The overall differences found between the commercial and smallholder flocks may be explained by differences in poultry husbandry practices and hatchery sources.     

Antimicrobial resistance in fecal generic Escherichia coli and Salmonella spp. obtained from Ontario sheep flocks and associations between antimicrobial use and resistance

The purpose of this study was to examine the prevalence and patterns of antimicrobial resistance (AMR) in enteric bacteria obtained from Ontario sheep flocks, and associations between antimicrobial use (AMU) and AMR. Forty-nine sheep producers participated for a 1-year interval between 2006 and 2008. Two-hundred and eighty-three pooled fecal samples were collected from the flocks during initial and final visits. Up to 3 isolates of Salmonella spp. and generic E. coli per pooled fecal sample were tested for susceptibility to 15 antimicrobials. Resistance was infrequent among Salmonella (0%, n = 7 isolates) and low among E. coli (13.1%; n = 849) isolates. A small number of isolates were resistant to antimicrobials classified as being of very high importance to human health. Tetracycline resistance was most frequently observed (12.0%). Logistic regression was used to model potential AMU (qualitative and quantitative) risk factors for tetracycline resistance in generic E. coli from final visits. Qualitative analysis indicated that the use of injectable sulfonamides [including trimethoprim-sulfonamide combinations (TMS)] and tetracycline in the feed and water were significantly associated with tetracycline resistance (OR = 2.6, P = 0.01; and OR = 4.8, P ≤ 0.01, respectively). Quantitative analysis also indicated that TMS exposure rate was significantly associated with tetracycline resistance, which varied depending on the exposure rate. The exposure rate of tetracycline in the feed and water was only significant after the removal of one influential flock, warranting further research examining flocks with higher tetracycline exposure rates. Although the prevalence of AMR in participating flocks was relatively low, risk factors for resistance were identified.     

Antimicrobial resistance in Campylobacter spp. isolated from Ontario sheep flocks and associations between antimicrobial use and antimicrobial resistance

The objectives of this study were to determine the prevalence of antimicrobial resistance (AMR) in faecal Campylobacter spp. from lambs and adult sheep and associations between antimicrobial use (AMU) and AMR. A total of 275 faecal samples collected during initial and final visits from 51 sheep flocks, including one feedlot, across southern Ontario were tested for the presence of Campylobacter spp. Campylobacter jejuni was detected in 52% (143/275) of the faecal samples, Campylobacter coli in 7% (19/275), Campylobacter lari in 1% (2/275) and 2% (4/275) were non-speciated Campylobacter. Broth microdilution was used to test antimicrobial susceptibility of 162 isolates to nine antimicrobials. Campylobacter jejuni isolates (n = 142) were resistant to tetracycline (39%), ciprofloxacin (4%), nalidixic acid (4%) and telithromycin (1%). C. coli isolates (n = 19) were resistant to tetracycline (74%), and azithromycin, clindamycin, erythromycin, and telithromycin (5%). The C. lari isolate displayed resistance to nalidixic acid. No statistically significant associations were found between AMU and AMR during multivariate modelling in this study.     

Antibiotic resistance in Escherichia coli and Enterococcus spp. isolates from commercial broiler chickens receiving growth-promoting doses of bacitracin or virginiamycin

Antibacterial agents such as zinc bacitracin (ZB) and virginiamycin (VG) are used as growth promoting agents (GP) in broiler chicken production. The objective of this study was to evaluate the effect of the use of ZB and VG on the emergence of antibacterial resistance in a commercial broiler chicken farm. Three trials were conducted using 3 different diets: one without antibacterial agents, one containing VG, and one with ZB. Escherichia coli and Enterococcus spp. strains were isolated and tested for their susceptibility to various antibacterial agents. The occurrence of the resistance genes vatD, ermB, and bcrR in Enterococcus spp. isolates was determined by polymerase chain reaction (PCR). Comparative quantification of vatD and bcrR genes in total deoxyribonucleic acid (DNA) extracts from litter was done by SYBR Green Real-Time PCR (QPCR). Escherichia coli and Enterococcus spp. isolates from diet groups had different levels of resistance to various antibacterial agents over time. These GPs did not select for specific antibacterial agent resistance (AAR) in Enterococcus spp. The use of GPs seemed to lower the percentage of E. coli isolates resistant to some antibacterial agents. The presence of the bcrR gene could not explain all resistant phenotypes to ZB. Genes other than vatD and ermB might be involved in the resistance to VG in Enterococcus spp. Use of GPs was not associated with presence of the bcrR gene in DNA extracts from litter, but use of VG was associated with vatD presence.     

Antimicrobial resistance in Enterococcus spp. and Escherichia coli isolated from poultry feed and feed ingredients

Poultry feed is at the start of the food safety chain in the "farm-to-fork" model, and might serve as a source of antimicrobial resistant bacteria present in poultry meat. Antimicrobial resistance was investigated in 1137 enterococci and 163 Escherichia coli strains recovered from 23 samples of commercial broiler feed and 66 samples of raw feeding materials taken over half a year timespan. Enumeration of enterococci and E. coli were also performed using traditional plating and fluorescent in situ hybridisation methods. Viable enterococci were detected in all feed samples and in 66% of samples of separate feed ingredients, while E. coli was present in 50% and 32% of feed and raw feeding materials, respectively. The median values (50th percentile) for plate and FISH counts for feeds were, respectively, 2.70 log CFU/g and 5.52 log cells/g for enterococci, and 0.15 log CFU/g and 6.00 log cells/g for E. coli. Among enterococci recovered from feed ingredients, resistance to rifampicin, erythromycin, nitrofurantoin, tetracycline, and ciprofloxacin was found in 59.8%, 21.6%, 21.2%, 18.0% and 6.9% of the isolates, respectively. A considerable proportion of the enterococci isolates obtained from broiler feed displayed resistance to tetracycline (69.1%), rifampicin (58.5%), erythromycin (52.9%) and nitrofurantoin (36.2%). Lower percentage of resistance was observed to chloramphenicol (4.6%), ciprofloxacin (3.9%), vancomycin (1.9%) and ampicillin (1.2%). Among E. coli recovered from feed ingredients and poultry feeds, resistance to ampicillin, tetracycline and streptomycin was found in 22.9%, 27.6% and 19.0% and in 22.4%, 41.4% and 17.0% of the isolates, respectively. These data show that feedstuffs and poultry feeds are extensively contaminated by resistant enterococci and, in a lesser extent, by E. coli, thus leading to their introduction in the farm environment.     

Associations between antimicrobial use and the prevalence of antimicrobial resistance in fecal Escherichia coli from feedlot cattle in western Canada

A randomized, controlled, blinded clinical trial was performed at a research feedlot in western Canada. Auction-market-derived steers (n = 288) were randomly assigned to 1 of 3 treatments: 1) no antimicrobials on arrival; 2) oxytetracycline in the starter ration for 14 d; and 3) long-acting oxytetracycline subcutaneously on day 0. Minimal inhibitory concentrations of 7 antimicrobials were determined for 3 generic fecal E. coli isolates per animal on arrival and throughout the feeding period. There was a low prevalence of antimicrobial resistance in generic E. coli isolates from calves on arrival. There were increased proportions of cattle with resistant E. coli isolates early in the feeding period among calves in groups 2 and 3. Individual animal treatments were not associated with increased proportions of cattle with resistant E. coli isolates preslaughter. There was no difference in the proportion of animals with E. coli isolates resistant to tetracycline between the treatment groups preslaughter. However, there were significantly more animals with tetracycline resistant isolates of E. coli preslaughter than at arrival.     

Heat resistance in liquids of Enterococcus spp., Listeria spp., Escherichia coli, Yersinia enterocolitica, Salmonella spp. and Campylobacter spp

The aim of the work was to collect, evaluate, summarize and compare heat resistance data reported for Campylobacter, Enterococcus, Escherichia, Listeria, Salmonella and Yersinia spp. The work was limited to resistance in liquids with pH values 6-8. Results obtained under similar experimental conditions were sought. Thermal destruction lines for the various bacterial groups studied were constructed using log10 D values and treatment temperatures. There was a good linear relationship between log10 D and temperature with Escherichia coli, listerias and salmonellas. For campylobacters, enterococci and yersinias the relationships were weaker but, nevertheless, present. Using the slopes of the lines and their 95% confidence limits, z values and their 95% confidence limits were calculated. z values were compared with z values obtained from reports. The equations for the lines were also used for calculation of predicted means of D values at various treatment temperatures. 95% confidence limits on predicted means of D values and on predicted individual D values were also calculated. Lines and values are shown in figures and tables. Differences in heat resistance noted between and within the bacterial groups studied are discussed.     

Prevalence of antimicrobial resistance and resistance genes in faecal Escherichia coli isolates recovered from healthy pets

Faecal samples of healthy dogs (n=39) and cats (n=36) obtained in Northern Portugal were seeded on Levine agar plates, and two Escherichia coli isolates per sample were recovered (78 of dogs and 66 of cats). The susceptibility to 16 antimicrobial agents was tested in this series of 144 E. coli isolates. Almost 20% of them showed tetracycline resistance and 12 and 15% presented ampicillin or streptomycin resistance, respectively. The percentage of resistance to the other antimicrobial agents was in all cases below 4%, and no resistant isolates were detected for ceftazidime, imipenem, cefoxitin or amikacin. Two isolates (from one dog) showed cefotaxime-resistance and harboured both the CTX-M-1 and OXA-30 beta-lactamases. A bla(TEM) gene was detected in 12 of 17 ampicillin-resistant isolates, the aac(3)-II gene in the three gentamicin-resistant isolates, aadA in 7 of 22 streptomycin-resistant isolates, and tet(A) and/or tet(B) gene in all 28 tetracycline-resistant isolates. The gene encoding class 1 integrase was detected in six E. coli isolates, including the four trimethoprim-sulfamethoxazole-resistant isolates and those two harbouring CTX-M-1 and OXA-30 beta-lactamases; different gene cassette arrangements were identified: dfrA1+aadA1 (two isolates), dfrA12+orfF+aadA2 (two isolates) and bla(OXA30)+aadA1 (two isolates). One amino acid change in GyrA protein (Ser83Leu or Asp87Tyr) was detected in four nalidixic acid-resistant and ciprofloxacin-susceptible isolates and two amino acid changes in GyrA (Ser83Leu+Asp87Asn) and one in ParC (Ser80Ile) were identified in one nalidixic acid- and ciprofloxacin-resistant isolate. Faecal E. coli isolates of healthy pets could be a reservoir of antimicrobial resistance genes.     

宠物源大肠杆菌耐药性调查

宠物细菌产生的耐药性不仅影响治疗效果,而且由于人类经常与小动物亲密频繁地接触,犬猫等身上的耐药菌非常有可能传给人类,导致严重的公共卫生问题[1].鉴于此,近年来欧美等发达国家对小动物临床的细菌耐药问题开始重视,已有不少宠物源细菌耐药性的研究[2-3].但在我们国家还没有这方面的认识,实际上随着国内宠物行业的发展,抗菌药的使用量非常大,由此导致的耐药问题可能比较突出.目前,国内已有大量关于畜禽病原菌耐药性的研究[4-5],但还未见有宠物源细菌耐药性方面的报道.本研究拟调查宠物源大肠杆菌对常用抗菌药物的敏感性,为小动物临床合理使用抗菌药物提供指导.     

Comparison of antimicrobial resistance in generic Escherichia coli and Salmonella spp. cultured from identical fecal samples in finishing swine

The objectives of this study were to investigate the associations between antimicrobial resistance patterns in generic Escherichia coli and Salmonella spp. isolates recovered from identical pen pooled fecal samples, and to evaluate potential clustering of multiple isolates of these organisms within identical fecal samples. Up to 5 generic E. coli (n = 922 isolates) and Salmonella spp. (n = 922 isolates) isolates were obtained from each of 188 pen pooled fecal samples that had been collected from 45 finishing swine farms in Alberta in 2000, and tested for susceptibility to 15 antimicrobials. No isolates of either organism were resistant to 3rd generation cephalosporins or fluoroquinolones, which in Canada are considered antimicrobials of very high importance to human health. Approximately twice as many generic E. coli isolates as Salmonella spp. isolates were resistant to at least 1 antimicrobial. In addition, E. coli isolates showed more multidrug-resistance patterns. No significant association was observed between the resistance phenotypes of Salmonella spp. and E. coli at the fecal sample level. More clustering at the sample level was observed for proportions of antimicrobial resistance (AMR) in Salmonella spp. isolates than E. coli indicating that in future studies it might be sufficient to test fewer than 5 Salmonella spp. isolates per sample.     

Characterization of integrons-mediated antimicrobial resistance among Escherichia coli strains isolated from bovine mastitis

To assess the prevalence of antimicrobial resistance and class I integrons in Escherichia coli strains (n=58) isolated from bovine mastitis in Inner Mongolia, antimicrobial susceptibility and the presence of various types of integrons were characterized. Most isolates were susceptible to amikacin, colistin, ceftazidime, gentamicin and kanamycin, while those also exhibited high resistant incidence rates to ampicillin, amoxicillin, sulfadiazine and sulfamethoxydiazine. The integrase gene of integrons was amplified by PCR using degenerate primers. The integrons were confirmed by restriction fragment length polymorphism (RFLP) analysis of positive PCR products. Neither class II nor class III integron was detected, while 56.90% (n=33) of the isolates were positive for the presence of intI1 gene. Sequencing analysis of gene cassettes revealed that seven gene cassettes were found, which encoded resistance to trimethoprim (dfrA1 and dfrA17), aminoglycosides (aacA4, aadA1 and aadA5) and chloramphenicol (catB3), respectively. Of them, the gene cassette array dfrA17-aadA5 was found most prevalent (62.96%). The percentage of positive-integron among the isolates whose resistant profile was relatively broad (n> or =7) is 100.00%, while the one in narrow-profile isolates (n=2-6) is 30.56%. The correlation analysis revealed the incidence of integrons among the isolates were highly related to the resistant profile, indicating integrons play an important role in the dissemination and spread of the antimicrobial resistant strains.     

Observations on production of hemolysin, heat-labile enterotoxin and antimicrobial drug resistance among enterotoxigenic Escherichia coli from pigs.

A total of 52 isolates of Escherichia coli belonging to enterotoxigenic serotypes from piglets with diarrhea were examined for hemolysis, production of cholera-like enterotoxin (LT) and susceptibility to 10 antimicrobial drugs. A strong association between production of LT and hemolysis was seen. Ninety percent of 29 hemolylic isolates were LT⁺ whereas 100% of 23 nonhemolytic isolates were LT⁻ in a commercial latex agglutination assay. Antimicrobial susceptibility tests employing disc diffusion showed that resistance to trimethoprim-sulfamethoxazole (TMS), neomycin and tetracycline was significantly less among LT⁺ isolates compared to LT⁻ ones. Enrofloxacin was the only antimicrobial drug to which all the 52 isolates were susceptible.     

西藏牦牛大肠杆菌耐药性调查

对西藏部分牧区牛源大肠杆菌进行耐药性考察,在对发病牦牛进行致病性大肠杆菌分离鉴定、纯化培养的基础上,采用美国NCCLS规定的纸片扩散法进行药敏试验,结果表明,分离培养的10株牛源大肠杆菌对头孢氨苄、头孢唑林、卡那霉素、阿米卡星、磷霉素、喹诺酮类药物的敏感率均为100%,可为临床合理选药、有效治疗牦牛大肠杆菌病提供依据。     

华南虎源大肠杆菌耐药性与致病性研究

为了解华南虎源大肠杆菌的耐药性与致病性情况,采集龙岩梅花山虎园老虎粪便分离大肠杆菌,并对分离株进行药敏试验、动物毒力试验以及毒力基因检测。结果表明,分离株对复方新诺明(100%)、多西环素(75%)和阿莫西林/克拉维酸(75%)产生耐药;对庆大霉素的敏感性较好;对头孢呋辛和头孢噻呋敏感性下降。动物毒力试验显示,分离株具有较强的致病性,同时毒力基因aat A的检出率为100%,irp2检出率为75%;papC、iucD、vat和iss检出率均为50%,tsh和cva/cvi未检出。虎源大肠杆菌对常用药物具有一定的耐药性,同时表现出较强的致病性,其毒力基因的携带率较高。该结果为兽医临床合理用药提供一定的科学依据。     

Antimicrobial resistance in Staphylococcus spp., Escherichia coli and Enterococcus spp. in dogs given antibiotics for chronic dermatological disorders, compared with non-treated control dogs

The aim of this study was to evaluate antimicrobial resistance in canine staphylococci, Escherichia coli and enterococci, which were isolated from 22 dogs with pyoderma and a history of previous antibiotic treatment, compared to bacterial isolates from 56 non-treated control dogs. Two isolates of each bacterial species per dog were investigated, if detected. Staphylococcal isolates from dogs with pyoderma (35 isolates) were more resistant to sulphatrimethoprim than the isolates from controls (56 isolates) (57% vs. 25%, p < 0.004). Multiresistance in staphylococci was also more common in dogs with pyoderma (29% vs. 9%, p = 0.02). A similar trend among isolates of E. coli was detected (24 and 74 isolates from treated and control dogs, respectively), but the differences were not significant. Resistance for macrolide-lincosamides was approximately 20% among staphylococci in both groups. Resistance to ampicillin among enterococci was 4%-7%. The age of the dogs might have an impact on resistance: multiresistance among staphylococcal isolates from younger dogs (< or = 5 years) was more common than in older dogs (26 years) (24%, vs. 0%, 63 and 27 isolates, respectively, p = 0.02). Staphylococci in younger dogs were more resistant to tetracycline (48% vs. 11%, p < 0.001) and sulphatrimethoprim (48% vs. 15%, p < 0.01) than those in older dogs. In contrast, the isolates of E. coli from older dogs tended to be more resistant, although a significant difference was detected only in resistance to tetracycline (13% vs. 2% of 40 and 50 isolates respecthely, p = 0.04)). The results of this small study indicate that resistance in canine staphylococci in the capital area of Finland is comparable with many other countries in Europe. Resistance in indicator bacteria, E. coli and enterococci, was low.     

Virulence-associated genes and antimicrobial resistance among avian pathogenic Escherichia coli from colibacillosis affected broilers in Pakistan

Avian pathogenic Escherichia coli (APEC) causes colibacillosis that leads to high morbidity and mortality among poultry birds. To date, there is a lack of knowledge about virulence-associated genes (VAGs) and multidrug resistance of APEC isolates from Pakistan. In this study, we determined the VAGs and antibiotic resistance profiles of APEC isolates recovered from colibacillosis affected broilers in Faisalabad region of Pakistan. A total of 84 diseased and dead birds from different local broilers farms were collected and examined for the gross lesions of colibacillosis by conducting postmortem examination. Of these, APEC isolates were recovered from 75 (89.2%) birds. Antibiotic susceptibility tests against 11 antimicrobial agents showed the highest resistance against ampicillin (98.6%) followed by tetracycline (97.3%) and ciprofloxacin (72%). The presence of 11 virulence-associated genes (VAGs) was detected by multiplex polymerase chain reaction (PCR). Of the 75 APEC, 32 (42.6%) harbored > 5 VAGs. Most commonly found genes were increased serum survival (iss; 84%), iron transport (iutA; 74.6%), and colicin V (ColV; 60%). Twenty-two isolates (29.3%) were found to possess a combination of VAGs; iss, tsh, iroN, and iutA, in addition to other VAGs. To the best of our knowledge, this is the first report on the detection of virulence-associated genes and multidrug resistance among APEC isolates in Pakistan. In the future, the strains with the predominant set of VAGs can be used for colibacillosis diagnosis and as a potential vaccine candidate.