Pathogenic and fecal Escherichia coil strains from turkeys in a commercial operation

The biochemical phenotypes and antimicrobial susceptibility patterns of 105 clinical Escherichia coli isolates from flocks with colibacillosis in a turkey operation were compared with 1104 fecal E. coli isolates from 20 flocks in that operation. Clinical isolates and 194 fecal isolates with biochemical phenotypes or minimum inhibitory concentrations for gentamicin and sulfamethoxazole similar to clinical isolates were tested for somatic antigens and the potential virulence genes hylE, iss, tsh, and K1. The predominant biochemical phenotype of clinical isolates contained 21 isolates including 14 isolates belonging to serogroup 078 with barely detectable beta-D-glucuronidase activity. Thirty-five fecal isolates had biochemical phenotypes matching common phenotypes of clinical isolates. Sixty-six (63%) clinical isolates exhibited intermediate susceptibility or resistance to gentamicin and sulfamethoxazole compared with 265 (24%) fecal isolates (P < 0.001). Seventy-seven clinical isolates reacted with O-antisera, of which 51 (66%) belonged to the following serogroups: O1, O2, O8, O25, O78, O114, and O119. In comparison, 8 of 35 (23%) fecal isolates subtyped on the basis of biochemical phenotype belonged to these serogroups and four of 167 (2%) fecal isolates subtyped on the basis of their antimicrobial resistance patterns belonged to these serogroups. Iss, K1, and tsh genes were detected more often among clinical isolates than these fecal isolates (P < 0.05). In summary, a small subgroup of E. coli strains caused most colibacillosis infections in this operation. These strains existed at low concentration in normal fecal flora of healthy turkeys in intensively raised flocks. The data suggest that colibacillosis in turkey operations may be due to endogenous infections caused by specialized pathogens.     

Antimicrobial resistance in fecal Escherichia coli isolated from growing chickens on commercial broiler farms

To investigate the effects of rearing practices of commercial broiler chickens on the incidence of antimicrobial resistance in commensal Escherichia coli isolates, fecal E. coli isolates obtained in 4 farms were screened for anitimicrobial resistance. Ten E. coli isolates were recovered from each of the fecal samples collected from 10 birds in the farms at the ages of 2 days, 14-17 days, and 47-50 days. In 2 out of the 4 farms, no antimicrobials were used during the rearing period. In the other two farms, following collection of the fecal samples at 14 and 15 days of age, oxytetracycline (OTC), sulfadimethoxine (SDMX), and tylosin were given to birds on one farm and SDMX was used in the other. Isolates resistant to ampicillin and OTC that were obtained from an untreated flock at different sampling times were closely related to each other by pulsed-field gel electrophoresis patterns (PFGE) of XbaI-digested chromosomal DNA. PFGE analysis together with in vitro conjugation experiments suggested that diversity of resistance phenotypes within a clone may be resulted from the acquisition and loss of R-plasmids in an untreated and a treated flock. The numbers of resistance phenotypes observed among fecal isolates increased during the growth of the chickens in all the farms. The results in the present study suggest that persistence of commensal E. coli strains resistant to antimicrobials even in the absence of antimicrobial administration. It is also hypothesized that horizontal transmission of resistance determinants resulted in the emergence of different resistance phenotypes in those farms.     

Associations among antimicrobial drug treatments and antimicrobial resistance of fecal Escherichia coli of swine on 34 farrow-to-finish farms in Ontario, Canada

Logistic regression was used to model associations between antimicrobial treatment and resistance among fecal Escherichia coli of finisher pigs at the farm level. Four sets of potential risk factors representing different levels of refinement of antimicrobial use on farms were modelled on resistance to antimicrobials. Final models for each antimicrobial were constructed from treatment and management variables significant on initial screening, and corrections for overdispersion were made. In general, in-feed antimicrobial treatment of pigs was more consistently associated with an increased risk of resistance than individual-animal treatment. Antimicrobial treatment in starter rations was significant in final models of resistance to ampicillin, carbadox, nitrofurantoin, sulfisoxizole, and tetracycline. Treatment in grower–finisher rations was significantly associated with resistance to ampicillin, spectinomycin, sulfisoxizole, and tetracycline. There was little evidence that in-feed antimicrobials increased the risk of resistance to gentamicin, which is a drug used only for individual-pig treatment in this study population. These results suggest that antimicrobial medication of rations of post-weaning pigs selects for and maintains antimicrobial resistance among E. coli of finisher pigs. Although resistance was common on farms that did not medicate rations of post-weaning pigs, the results indicate that antimicrobial use does increase the risk of resistance to the antimicrobials studied.     

Antimicrobial susceptibility of Escherichia coli isolates from dairy farms using organic versus conventional production methods

OBJECTIVE: To compare antimicrobial susceptibility patterns of Escherichia coli isolates cultured from fecal samples from cows and calves on dairy farms that used organic (ie, no or severely limited antimicrobial use) versus conventional production methods. DESIGN: Cross-sectional study. SAMPLE POPULATION: Fecal samples from 10 cows and 10 calves on each of 30 organic dairy farms and 30 neighboring conventional dairy farms in Wisconsin. PROCEDURE: E. coli isolates obtained from the fecal samples were tested for susceptibility to 17 antimicrobials by means of a microbroth dilution test. Prevalence of antimicrobial resistance was compared between organic and conventional dairy farms. RESULTS: E. coli was isolated from 1,121 (94%) fecal samples. Farm type (organic vs conventional) and animal age (cow vs calf) were significantly associated with odds that E. coli isolates would be resistant to various antimicrobials. After controlling for age, logistic regression analyses indicated that isolates from conventional dairy farms had significantly higher rates of resistance to ampicillin, streptomycin, kanamycin, gentamicin, chloramphenicol, tetracycline, and sulfamethoxazole than did isolates from organic dairy farms. However, no significant differences were detected for the 10 other antimicrobials that were tested. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that compared with isolates from conventional dairy farms, E. coli isolates from organic dairy herds have significantly lower prevalences of resistance to 7 antimicrobials; however, prevalence of resistance was not significantly different for 10 other antimicrobials. Resistance was more common for isolates from calves than for isolates from adult dairy cows.     

Prevalence and patterns of antimicrobial resistance of fecal Escherichia coli among pigs on 47 farrow-to-finish farms with different in-feed medication policies in Ontario and British Columbia

The main objectives of this study were to determine the prevalence and patterns of antimicrobial resistance in pigs on farms that medicated swine ration and those that did not. A total of 940 isolates of Escherichia coli from 188 pooled fecal samples obtained from weaner and finisher pigs on 47 farrow-to-finish swine farms (34 farms used in-feed medication and 13 did not) were tested for susceptibility to 21 antimicrobials using a breakpoint concentration method. The prevalence of resistance varied widely (0.0% to 81.3%) among the antimicrobials tested. Ninety percent of all the isolates tested were resistant to one or more antimicrobials. The most common multi-drug resistance patterns were to 2 to 6 antimicrobials. Resistance was significantly more frequent (P < 0.01) on farms that used in-feed medication compared to those that did not, and significantly more frequent (P < 0.01) in weaner pigs compared to finisher pigs. These findings indicate that resistance to a broad range of antimicrobials was prevalent among fecal E. coli isolates of pigs on study farms, and that this constitutes a potential reservoir for resistance genes that could spread to pathogens. The findings also provide further evidence that use of medication in swine rations provides selective pressure for antimicrobial resistance in E. coli in pigs.     

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.     

Prevalence and pattern of antimicrobial susceptibility in Escherichia coli isolated from pigs reared under antimicrobial-free and conventional production methods

OBJECTIVE: To determine and compare levels and patterns of antimicrobial resistance among Escherichia coli isolated from pigs on farms that did not use antimicrobial agents versus pigs produced under conventional methods. DESIGN: Cross-sectional study. Sample Population-35 antimicrobial-free and 60 conventional swine farms. PROCEDURES: Farms were visited once, and fecal samples were collected from 15 finisher pigs if available. One E coli isolate from each sample was tested for susceptibility pattern to 14 antimicrobial agents by use of microbroth dilution. RESULTS: E coli isolates were recovered from 1,381 (97.1%) of 1,422 fecal samples. Herd size was significantly larger for conventional swine farms. Resistance to ceftriaxone, ciprofloxacin, or nalidixic acid was not observed on any of the 95 farms. Three isolates from 2 conventional farms were resistant to ceftiofur. Conventional farms had significantly higher levels of resistance to ampicillin, sulfamethoxazole, tetracycline, and chloramphenicol, compared with antimicrobial-free farms. Fourteen percent of E coli isolates were susceptible or had intermediate resistance to all the tested antimicrobial agents. The 3 most frequent patterns of multiple resistance were streptomycin-tetracycline, sulfamethoxazole-tetracycline, and kanamycin-streptomycin-sulfamethoxazole-tetracycline. CONCLUSIONS AND CLINICAL RELEVANCE: Cessation of antimicrobial use did not appear to result in an immediate reduction in antimicrobial resistance in swine farms. Prospective studies of long-term antimicrobial usage and cessation are needed to estimate the extent to which food animal production may be contributing to antimicrobial drug resistance and might provide a direct measure of the rates of reversibility of antimicrobial drug resistance that might be achieved by curtailing antimicrobial usage.     

Effects of feeding wet corn distillers grains with solubles with or without monensin and tylosin on the prevalence and antimicrobial susceptibilities of fecal foodborne pathogenic and commensal bacteria in feedlot cattle

Distillers grains, a coproduct of ethanol production from cereal grains, are composed principally of the bran, protein, and germ fractions and are commonly supplemented in ruminant diets. The objective of this study was to assess the effect of feeding wet distillers grains with solubles (WDGS) and monensin and tylosin on the prevalence and antimicrobial susceptibilities of fecal foodborne and commensal bacteria in feedlot cattle. Cattle were fed 0 or 25% WDGS in steam-flaked corn-based diets with the addition of no antimicrobials, monensin, or monensin and tylosin. Fecal samples were collected from each animal (n = 370) on d 122 and 136 of the 150-d finishing period and cultured for Escherichia coli O157. Fecal samples were also pooled by pen (n = 54) and cultured for E. coli O157, Salmonella, commensal E. coli, and Enterococcus species. Antimicrobial resistance was assessed by determining antimicrobial susceptibilities of pen bacterial isolates and quantifying antimicrobial resistance genes in fecal samples by real-time PCR. Individual animal prevalence of E. coli O157 in feces collected from cattle fed WDGS was greater (P < 0.001) compared with cattle not fed WDGS on d 122 but not on d 136. There were no treatment effects on the prevalence of E. coli O157 or Salmonella spp. in pooled fecal samples. Antimicrobial susceptibility results showed Enterococcus isolates from cattle fed monensin or monensin and tylosin had greater levels of resistance toward macrolides (P = 0.01). There was no effect of diet or antimicrobials on concentrations of 2 antimicrobial resistance genes, ermB or tetM, in fecal samples. Results from this study indicate that WDGS may have an effect on the prevalence of E. coli O157 and the concentration of selected antimicrobial resistance genes, but does not appear to affect antimicrobial susceptibility patterns in Enterococcus and generic E. coli isolates.     

Associations of antimicrobial uses with antimicrobial resistance of fecal Escherichia coli from pigs on 47 farrow-to-finish farms in Ontario and British Columbia

This study assessed the associations between antimicrobial use and other management practices in pigs and antimicrobial resistance in generic Escherichia coli recovered from feces of weaner and finisher pigs on 39 purposefully selected farrow-to-finish farms in Ontario and 8 in British Columbia. Antimicrobials (n = 13), most frequently penicillins and tetracycline, wer dministered to different age groups of pigs on study farms through various routes of administration. Logistic regression was used to model risk factors to antimicrobial resistance in fecal E. coli of pigs for the following antimicrobials: ampicillin, apramycin, carbadox, cephalothin, chloramphenicol, kanamycin, neomycin, nitrofurantoin, spectinomycin, streptomycin, sulfamethoxazole tetracycline, and cotrimoxazole (trimethoprim and sulfamethoxazole). Use of antimicrobials in weaner pigs compared with use in finisher pigs was associated with resistance in most models. There was phenotypic evidence of different mechanisms of resistance selection, including direct selection [use of carbadox was associated with carbadox resistance (OR = 6.48)]; cross-resistance [use of spectinomycin was associated with streptomycin resistance (OR = 2.29)]; and possible co-selection [ceftiofu use was associated with tetracycline resistance (OR = 6.12)]. These results provide further evidence that use of antimicrobials in pigs selects for resistance among fecal E. coli within and between classes of antimicrobials.     

Antimicrobial resistance in generic Escherichia coli isolated from swine fecal samples in 90 Alberta finishing farms

The objective of this study was to determine the prevalence of antimicrobial resistance in generic Escherichia coli isolates obtained from 90 Alberta finisher swine farms. Up to 5 isolates were obtained from each of 269 pooled fecal samples and were classified as susceptible or resistant according to Clinical and Laboratory Standards Institute guidelines. Of the 1322 isolates, 166 (12.6%) were susceptible to all 15 antimicrobials. No resistance to amikacin, ceftiofur, ceftriaxone, or ciprofloxacin, antimicrobials of importance in human medicine, was observed. Relatively low frequencies of resistance were observed to gentamicin (1.1%), amoxicillin/clavulanic acid (0.7%), and cefoxitin (0.7%). Higher frequencies of resistance were observed for tetracycline (78.9%), sulfisoxazole (49.9%), streptomycin (49.6%), ampicillin (30.6%), chloramphenicol (17.6%), kanamycin (10%), and trimethoprim/ sulfamethoxazole (6.4%). Among the isolates resistant to > or = 2 antimicrobial classes, 20.8%, 20.6%, 18.2%, 7.0%, 1.8%, 0.2%, and 0.2% were resistant to 2, 3, 4, 5, 6, 7, and 8 antimicrobials, respectively. The most common multidrug-resistance patterns (resistance to > or = 2 antimicrobial classes) were streptomycin-tetracycline (9.4%), streptomycin-sulfisoxazole-tetracycline (6.2%), and ampicillin-streptomycin-sulfisoxazole-tetracycline (6.1%). More clustering (higher intra-class correlation coefficients) in antimicrobial resistance was observed for isolates at the same visit than for isolates from different visits in the same farm, indicating that sampling more farms, testing fewer isolates per visits, and taking longer periods between visits may be appropriate and more efficient for a better understanding of potential shifts in resistance over time.     

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.     

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。     

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.     

Identification, antimicrobial resistance profiles, and virulence of members from the family Enterobacteriaceae from the feces of yellow-headed blackbirds (Xanthocephalus xanthocephalus) in North Dakota

Public pressure to reduce or eliminate antimicrobials as ingredients of feed for poultry and other agricultural animals is mounting, primarily due to the fear of multidrug-resistant bacteria in clinical infections in both animals and humans. Exploration of the occurrence of antibiotic resistance in the gut flora of wildlife avian flocks that presumptively do not receive antimicrobials will determine the rate of resistance in a na?ve population. Fecal samples collected from a healthy population of the yellow-headed blackbirds (YHB) (Xanthocephalus xanthocephalus) in North Dakota were cultured to determine what genera and species of gram-negative facultative anaerobic bacteria these wild birds carry in their intestinal flora and to evaluate the antimicrobial susceptibility profiles. Isolates of Escherichia coli were further characterized for the presence of putative virulence factors and for pathogenic potential using the chicken embryo lethality assay (ELA). The ELA was performed in chicken embryos with challenges at both 12 days and 16 days of incubation to determine whether the 16-day-old embryos were better able to fight the infection and subsequent disease and also to determine whether the ELA could distinguish between primary and secondary avian Escherichia coli pathogens. After screening 33 isolates from the 21 fecal samples, only two E. coli isolates were identified. The predominant genus and species of bacterium identified was Pantoea agglomerans. Collectively, 12 of the 33 isolates (36%) exhibited no resistance to any antimicrobial tested. However, several multidrug-resistant isolates of varying genera were identified. Among the antimicrobial resistances observed, the most common was to ampicillin (60%), followed by cephalothin (33%). Neither E. coli isolate belonged to serogroups that are notorious for causing major outbreaks of colibacillosis in poultry, and only one E. coli isolate retained resistance to any antibiotics; nevertheless, the ELA results indicate that at least one of these E. coli may be a primary pathogen of chickens. This study demonstrates that antibiotic resistance occurs in the gut flora of natural populations of YHB despite the absence of antibiotic pressure. In addition, these results indicate that YHB will harbor E. coli isolates that are potentially pathogenic in poultry. However, these E. coli isolates are not a significant reservoir for multiple antibiotic resistances nor are they widespread in the population of YHB surveyed in North Dakota.     

Antimicrobial Activity of Tulathromycin and 14 Other Antimicrobials Against Virulent Rhodococcus equi In Vitro

This study determined the antimicrobial activity of tulathromycin against Rhodococcus equi in vitro. Ninety-eight virulent isolates of R. equi from equine clinical cases were examined, of which 20 isolates were macrolide resistant. A custom 96-well antimicrobial susceptibility testing plate was used, allowing 14 additional antimicrobials to be tested against R. equi. Isolates were cultured with various concentrations of antimicrobials, and minimal inhibitory concentration (MIC) values were determined. Tulathromycin was found to have poor activity in vitro against R. equi isolates susceptible or resistant to macrolides, with MIC50 and MIC90 values >64 ug/mL for all isolates. MIC values for other macrolides tested were similar to previously published data.     

Prevalence of antimicrobial resistance in fecal generic Escherichia coil isolated in western Canadian beef herds. Part II--cows and cow-calf pairs.

The objective of this study was to describe antimicrobial resistance in fecal generic Escherichia coli isolated from cows and cow-calf pairs from western Canadian beef herds. Susceptibility testing was completed on 1555 E. coli isolates (n = 533 cows from 69 herds) harvested from fresh fecal samples in the spring of 2002, and 630 isolates (n = 105 cow-calf pairs from 10 herds) collected in the spring of 2003. Only 1 cow isolate was resistant to an antimicrobial classified by Health Canada as being of very high importance to human medicine. Resistance to at least 2 antimicrobials was detected in 7.1% of the 2002 cow isolates, in 3.4% of the 2003 cow isolates, and 23.2% of the 2003 calf isolates. In the cows, resistance to at least 1 antimicrobial was not associated with cow breed (P = 0.16), cow age (P = 0.14), or previous cow treatment (P = 0.56). In the calves, resistance to at least 1 antimicrobial was not predicted by whether or not its dam was resistant to at least 1 antimicrobial (P = 0.36).     

Prevalence of resistance to 11 antimicrobials among Campylobacter coill isolated from pigs on 80 grower-finisher farms in Ontario.

We carried out a cross-sectional study to investigate antimicrobial resistance patterns of Campylobacter coli isolated from Ontario grower-finisher pigs. From January to June 2004, 1200 samples were collected from 80 farms by obtaining a constant number (15) of fecal samples per farm. Susceptibility of the isolates to 11 antimicrobial drugs was determined by the agar-dilution technique. The overall prevalence of resistance to 1 or more antimicrobials among the isolates was 99.2%. High levels of resistance were observed for azithromycin, clindamycin, erythromycin, streptomycin, and tetracycline: 91.7%, 82.5%, 81.4%, 70.7%, and 63.7%, respectively. For sulfamethoxazole, ampicillin, and nalidixic acid, resistance was observed in 40.3%, 26.6%, and 22.7% of the isolates, respectively. Although at very low levels, resistance was observed for ciprofloxacin (a fluoroquinolone), chloramphenicol, and gentamicin: in 2.4%, 1.7%, and 0.2%, respectively. Many of the isolates (29.7%) were resistant to 5 antimicrobials, the most common being azithromycin, clindamycin, erythromycin, streptomycin, and tetracycline. Isolates from the same farm showed at least 5 patterns of resistance. Results from this study indicate high levels of resistance to the antimicrobial drugs most commonly used in the Canadian swine industry (macrolides, lincosamides, and tetracyclines) among C. coli isolated from grower-finisher pigs in Ontario. Macrolides and fluoroquinolones are the drugs most commonly used to treat severe human campylobacteriosis. Fortunately, at present, there is little resistance to fluoroquinolones among C. coli from pigs in Ontario.     

Microarray based comparative genotyping of gentamicin resistant Escherichia coli strains from food animals and humans

Recent data from the European and Hungarian Antimicrobial Resistance Monitoring Systems have indicated that the routine use of gentamicin in human and veterinary medicine frequently leads to the selection of gentamicin resistance in Escherichia coli. The aim of this study was to provide molecular characterization of gentamicin resistance in clinical and commensal E. coli strains representing humans and food producing animals by genotyping for antimicrobial resistance and virulence using a miniaturized microarray. All 50 strains tested proved to be multidrug resistant defined as resistance to three or more antimicrobial classes. Antimicrobial resistances genes such as aadA1-like, strB, bla(TEM), sul1 and tet(A) or tet(B), and corresponding phenotypes (streptomycin-, ampicillin-, sulfamethoxazole- and tetracycline resistance) were detected in >50% of isolates regardless of the host or clinical background. However, certain genes encoding gentamicin resistance such as aac(6')-Ib and ant(2″)-Ia as well as catB3-like genes for phenicol resistance were only detected in human isolates. Among virulence genes, the increased serum survival gene iss was predominant in all host groups. Although the majority of gentamicin resistant E. coli strains were characterized by diverse antimicrobial resistance, and virulence gene patterns, accentuated links between catB3-like, aac(6')-Ib, bla(CTX-M-1) and sat genes could be detected in human strains. Further resistance/virulence gene associations (tet(A) with iroN and iss) were detected in poultry strains. In conclusion, the simultaneous characterization of antimicrobial resistance and virulence genotypes of representative clinical and commensal strains of E. coli should be useful for the identification of emerging genotypes with human and or animal health implications.     

Antimicrobial-induced endotoxin and cytokine activity in an in vitro model of septicemia in foals

OBJECTIVE: To determine which antimicrobials that are used to treat neonatal foals with septicemia attributable to Escherichia coli will minimize endotoxin release from bacteria and subsequent activity of inflammatory mediators while maintaining bactericidal efficacy. SAMPLE POPULATION: Blood samples from 10 healthy foals. PROCEDURE: Escherichia coli isolates A and B were isolated from 2 septicemic foals, and minimal inhibitory concentrations (MIC) were determined for 9 antimicrobials. Five of these antimicrobials were tested in vitro at 2 and 20 times their respective MIC. Whole blood or mononuclear cells grown in tissue-culture media were incubated with 105 colony-forming units of E. coli and each antimicrobial or saline (0.9% NaCl) solution. After 6 hours, number of viable bacteria remaining was determined, and supernatant was tested for endotoxin and tumor necrosis activity. RESULTS: Testing in whole blood was compromised by bactericidal effects of the blood itself. In mononuclear cell suspensions, each antimicrobial significantly reduced the number of viable bacteria to low or undetectable amounts. Antimicrobials did not differ significantly in efficacy of bacterial killing. Amikacin used alone or in combination with ampicillin resulted in significantly less endotoxin activity than did ampicillin, imipenem, or ceftiofur alone. There was a correlation between TNF-alpha and endotoxin activity. CONCLUSIONS AND CLINICAL RELEVANCE: Aminoglycosides appear less likely to induce endotoxemia and TNF-alpha synthesis during bactericidal treatment of E. coli septicemia, compared with beta-lactam antimicrobials. Use of ampicillin, imipenem, or ceftiofur in the treatment of septicemic neonatal foals should be accompanied by appropriate treatment for endotoxemia.