Antimicrobial resistance and genetic profiling of Escherichia coli from a commercial beef packing plant

The objective of this study was to investigate the extent of antimicrobial resistance and to genetically characterize resistant Escherichia coli recovered from a commercial beef packing plant. E. coli isolates were recovered by a hydrophobic grid membrane filtration method by direct plating on SD-39 medium. A total of 284 isolates comprising 71, 36, 55, 52, and 70 isolates from animal hides, washed carcasses, conveyers, beef trimmings, and ground beef, respectively, were analyzed. The susceptibility of E. coli isolates to 15 antimicrobial agents was evaluated with an automated broth microdilution system, and the genetic characterization of these isolates was performed by the random amplified polymorphic DNA (RAPD) method. Of the 284 E. coli isolates, 56% were sensitive to all 15 antimicrobial agents. Resistance to tetracycline, ampicillin, and streptomycin was observed in 38, 9, and 6% of the isolates, respectively. Resistance to one or more antimicrobial agents was observed in 51% of the E. coli isolates recovered from the hides but in only 25% of the E. coli from the washed carcasses. Resistance to one or more antimicrobial agents was observed in 49, 50, and 37% of the isolates recovered from conveyers, beef trimmings, and ground beef, respectively. The RAPD pattern data showed that the majority of resistant E. coli isolates were genetically diverse. Only a few RAPD types of resistant strains were shared among various sample sources. The results of this study suggest that antimicrobial-resistant E. coli isolates were prevalent during all stages of commercial beef processing and that considerably higher numbers of resistant E. coli were present on conveyers, beef trimmings, and ground beef than on dressed carcasses. This stresses the need for improving hygienic conditions during all stages of commercial beef processing and meatpacking to avoid the risks of transfer of antimicrobial-resistant bacteria to humans.     

Antimicrobial resistance observed in Escherichia coli strains isolated from fecal samples of cattle and pigs in Korea during 2003-2004

A total of 744 Escherichia coli strains isolated from 830 fecal samples of healthy cattle and pigs in all provinces of Korea were examined for resistance to 16 antimicrobials. The most frequently observed resistance in cattle isolates was to tetracycline (30.5%), followed by resistance to streptomycin (20.4%), ampicillin (12.0%) and chlorampenicol (6.9%). Prevalences of resistance to the same four antimicrobials in swine isolates were 96.3%, 66.8%, 66.1%, and 47.6%, respectively. The prevalence of resistance in pigs was much higher than that in cattle, with 98.3% of pig isolates and 37.1% of cattle isolates showing resistance to one or more of the antimicrobial agents tested.     

Antimicrobial resistance in Escherichia coli isolated from retail milk-fed veal meat from Southern Ontario, Canada

This study estimated the prevalence of Escherichia coli isolates in fresh retail milk-fed veal scallopini pieces obtained from grocery stores in Ontario, Canada. In addition, the prevalence and antimicrobial resistance patterns were examined for points of public health significance. One hundred fifty-three milk-fed veal samples were collected over the course of two sampling phases, January to May 2004 and November 2004 to January 2005. E. coli isolates were recovered from 87% (95% confidence interval, 80.54 to 91.83%) of samples, and antimicrobial susceptibility testing was conducted on 392 isolates. The prevalence of resistance to one or more antimicrobials was 70% (274 of 392), while the resistance to five or more antimicrobials was 33% (128 of 392). Resistance to ceftiofur (2.8%), ceftriaxone (3.6%), nalidixic acid (12%), and ciprofloxacin (3.8%) alone or in combination was observed. Eighty-five resistance patterns were observed; resistance to tetracycline only (7.4%) was observed most frequently. Individual antimicrobial resistance prevalence levels were compared with grain-fed veal and retail beef data from samples collected in Ontario. In general, resistance to individual antimicrobials was observed more frequently in E. coli isolates from milk-fed veal than in isolates from grain-fed veal and beef. Resistance to one or more antimicrobials and to five or more antimicrobials in E. coli isolates was more frequent in isolates from milk-fed veal than in isolates from grain-fed veal and beef. This study provides baseline data on the occurrence of resistance in E. coli isolates from milk-fed veal that can be compared with data for other commodities. Additionally, E. coli resistance patterns may serve as an indicator of antimicrobial exposure.     

Antimicrobial resistance and association with class 1 integrons in Escherichia coli isolated from turkey meat products

The objective of this study was to quantify the role of class 1 integrons in antimicrobial resistance in Escherichia coli isolated from turkey meat products purchased from retail outlets in the Midwestern United States. Of 242 E. coli isolates, 41.3% (102 of 242) tested positive for class 1 integrons. A significant association was shown between presence of class 1 integrons in E. coli isolates and the resistance to tetracycline, ampicillin, streptomycin, gentamicin, sulfisoxazole, and trimethoprim-sulfamethoxazole. Attributable risk analysis revealed that for every 100 E. coli isolates carrying class 1 integrons, resistance was demonstrated for ampicillin (22%), gentamycin (48%), streptomycin (29%), sulfisoxazole (40%), trimethoprim-sulfamethoxazole (7%), and tetracycline (26%). Non-integron-related antimicrobial resistance was demonstrated for ampicillin (65%), gentamycin (16.9%), streptomycin (42.1%), sulfisoxazole (35.8%), and tetracycline (49.7%). Population-attributable fraction analysis showed that class 1 integrons accounted for the following resistances: gentamycin, 71% (50 of 71), amoxicillin-clavulanic acid, 19.6% (6 of 33), nalidixic acid, 34% (7 of 21), streptomycin, 28% (30 of 107), sulfisoxazole, 38% (40 of 106), and tetracycline, 14%, (26 of 185). In conclusion, although class 1 integrons have been implicated in resistance to antimicrobial agents, other non-integron resistance mechanisms seem to play an important part.     

Antimicrobial resistance of Vibrio parahaemolyticus and Vibrio alginolyticus strains isolated from farmed fish in Korea from 2005 through 2007

The antimicrobial resistance patterns to 15 antimicrobial agents of Vibrio parahaemolyticus and Vibrio alginolyticus isolated from farmed fishes, including olive flounder (Paralichthys olivaceus), black rockfish (Sebastes schlegeli), red sea bream (Pagrus major), and sea bass (Lateolabrax japonicus), were investigated from 2005 through 2007. A total of 218 V. parahaemolyticus isolates and 153 V. alginolyticus isolates were obtained from the 180 fish samples collected from fish farms located along the southern coast of Korea. We found that 65.1% of V. parahaemolyticus and 85.6% of V. alginolyticus isolates showed antimicrobial resistance against more than one antimicrobial agent. The prevalence of resistance in V. parahaemolyticus isolates to ampicillin was highest (57.8%), followed by resistance to rifampin (11.9%), streptomycin (8.7%), and trimethoprim (6.4%). V. alginolyticus isolates were also most resistant to ampicillin (75.2%), followed by tetracycline (15.0%), trimethoprim (12.4%), and rifampin (9.8%). The prevalence of multiresistance to four or more antimicrobials was higher in V. alginolyticus (11.1%) than in V. parahaemolyticus (5%). Antimicrobial resistance rates per isolate of V. parahaemolyticus and V. alginolyticus possessing virulence genes were not different from those of the rest of the isolates.     

多重PCR法检测与分析鱼塘生态系统大肠杆菌的耐药基因与整合子

了解在鱼塘生态系统中,大肠杆菌耐药基因、整合酶基因的携带情况,并对其进行分析,可以为控制细菌耐药基因传播提供科学的依据。本文对来自鱼塘生态系统的165株大肠杆菌进行了检测,发现大肠杆菌中普遍带有耐药基因,携带有耐药基因的比率为91．51％。携带有2种耐药基因以上菌株的比率为86．06％,大肠杆菌耐药基因的携带以多重耐药基因为主。进而对带有4种以上耐药基因的菌株检测,I整合子酶基因阳性检出率为87．84％,携带有7种耐药基因的菌株全部为I整合酶基因阳性;11型整合酶基因阳性菌株检出率10．81％;1株为阴性;检测中没发现有携带Ⅲ型整合酶基因菌株。可见I整合子介导的耐药机制占主导位置,并且通过整合子的整合作用向周围环境扩散。     

Antibiotic resistance and virulence genes in commensal Escherichia coli and Salmonella isolates from commercial broiler chicken farms

Antibiotic resistance patterns and the presence of antibiotic and virulence determinants in 74 sorbitol-negative Escherichia coli and 62 Salmonella isolates from nine different broiler chicken farms were investigated. Each farm was supplied by one of three companies that used different antimicrobial agents in feed for growth promotion. The isolates were identified by API 20E for E. coli and by serological tests for Salmonella. The susceptibility of the isolates to antibiotics was determined by Sensititre using the Clinical and Laboratory Standards Institute's breakpoints. Fifty-two E. coli isolates (70.3%) and nine Salmonella isolates (14.52%) were multiresistant to at least nine antibiotics. The multiresistant isolates were evaluated for the presence of tetracycline resistance, integron class 1, and blacMY 2 genes by PCR. Of the 74 E. coli isolates, 55 were resistant to amoxicillin and ceftiofur. Among these 55 resistant E. coli isolates, 45 (81.8%) and 22 (40.0%) were positive for blacMY-2 and qacEdeltal-Sull genes, respectively. Tetracycline resistance was found in 56 isolates (75.8%) among which 12 (21.4%) and 24 (42.9%) gave positive results for tetA and tetB, respectively. Virulence genes (iss, tsh, and traT), aerobactin operon (iucC), and the eaeA gene were detected in some E. coli strains. Among the 27 amoxicillin- and ceftiofur-resistant Salmonella isolates, the blacMY-2 gene was detected in 22 isolates. The class 1 integron gene (qacEdeltal-Sull) was not detected in any Salmonella isolates, whereas the invasin (inv) and virulence (spy) genes were found in 61 (98.4%) and 26 (42%) of the Salmonella isolates, respectively. This study indicated that multiple antibiotic-resistant commensal E. coli and Salmonella strains carrying virulence genes can be found on commercial broiler chicken farms and may provide a reservoir for these genes in chicken production facilities. Except for the presence of tetB, there was no significant effect of feed formulations on the phenotypic or genotypic characteristics of the isolates.     

Evaluation of antibacterial resistance in Vibrio strains isolated from imported seafood and Italian aquaculture settings

Ninety-two Vibrio strains isolated over a period of 9 years from different sources (national and imported fishery products, shellfish, seawater from aquaculture settings, etc.) and belonging mostly to two species relevant for human health and fish pathology, V. parahaemolyticus and V. alginolyticus, were tested for resistance to different antibacterials using the standard disk diffusion test (five antibacterials: ampicillin, sulfamethoxazole, tetracycline, trimethoprim/sulfamethoxazole and chloramphenicol) and the Minimum Inhibitory Concentrations method (six antibacterial families: β-lactams, tetracyclines, chloramphenicol, aminoglycosides, quinolones, trimethoprim/sulfamethoxazole) providing consistent results. The analysis performed by standard disk diffusion test showed resistance to ampicillin in 82% of the strains, whereas 7% were resistant to sulfamethoxazole, 3% to tetracycline, and 1% to trimethoprim/sulfamethoxazole; no strain was found to be resistant to chloramphenicol. The results obtained by the MIC tests, because of the absence of established breakpoints for some antibacterials, were not readily interpretable for all substances, but showed no statistically significant difference to the results obtained by the standard disk diffusion test. Beside these results, high MIC₉₀ values (128 μg ml⁻¹ and above) were obtained for kanamycin and streptomycin, indicating that high concentrations of these antibacterials were required to inhibit the growth of the strains. Strains exhibiting resistance or intermediate resistance to two or more antibacterials represented 15% of the total and included, beside resistance to ampicillin, resistance to sulfamethoxazole or, for V. parahaemolyticus isolates, tetracycline. Interestingly, differences in the mechanism of resistance to β-lactams were detected between the species V. parahaemolyticus and V. alginolyticus.     

Antimicrobial resistance of Escherichia coli O157 from cattle in Korea

Of 45 Escherichia coli O157 isolates from cattle feces, which were collected between May 2000 and September 2003 in Korea, 32 were resistant to at least 1 antibiotic and 28 were resistant to 4 or more antibiotics, with 32, 30 and 30 of the isolates being resistant to streptomycin, tetracycline and sulfisoxazole, respectively. Two isolates were resistant to fluoroquinolones and to 10 or more of the 22 other antimicrobial agents that were tested. Thirteen antimicrobial resistant patterns were observed. The most frequent resistance type, which was found for 11 isolates, was streptomycin-tetracycline-kanamycin-ampicillin-piperacillin-cephalothin-sulfisoxazole-ticarcillin. Polymerase chain reaction (PCR) analysis of the isolates for E. coli O157 virulence markers revealed that 25 of the resistant E. coli O157 isolates tested positive for stx2 or both stx1 and stx2 genes. These findings suggest that many of the resistant E. coli O157 isolates might cause disease in humans.     

市售生禽类产品分离大肠埃希氏菌药物敏感分析

目的 生禽类产品中大肠埃希氏菌是细菌中最常见的传染病菌之一,耐药大肠埃希氏菌的生长和繁殖中能通过食物链将耐药性传递给人源致病菌,对人类健康的造成严重影响。方法 本文对市售生禽类产品中大肠埃希氏菌进行分离和鉴定,并通过抗生素最小抑制浓度确定分离菌株药敏谱。结果：从65份样本中分离得到55株大肠埃希氏菌,分离率高达84.46%;药敏结果显示,分离菌对亚胺培南(98.2%)、头孢西丁(92.73%)、头孢他啶(90.9%)、多粘菌素(90.9%)耐药率较高,对四环素最敏感(72.2%)。表型不同的大肠埃希氏菌分为典型菌株(45株)和非典型菌株(10株)药敏谱无显著性差异(P>0.5)。受试菌100%携带3种以上抗生素,有83.6%受试菌同时携带4种高耐药率的抗生素(亚胺培南、头孢西丁、头孢他啶、多粘菌素)。结论 市售生禽类产品中存在非常严重的多重耐药性大肠埃希氏菌污染,本文对大肠埃希氏菌的药性分析,旨在为临床预防治疗提供相应的理论参考。     

北疆部分地区食源性大肠杆菌血清型、毒力基因及耐药基因的研究

目的研究北疆部分地区食源性大肠杆菌的优势血清型、毒力基因和耐药基因的相关性。方法采用玻片凝集法测定了大肠杆菌血清型分布,PCR方法调查11种耐药基因和9种毒力基因。结果血清试验,定型菌株26株,分别属于17种血清型,其中O1、O115为优势血清型;PCR结果表明,不含有所检测耐药基因的分离株占12.50%,至少含有2种以及以上的分离株占50.00%,分离株对blaOXA基因携带率高达57.14%;检出率较高的毒力基因为fimC(64.29%)和fimA(25.00%)。结论 O1、O115为主要的血清型,2种血清型菌株拥有的耐药基因谱和毒力基因谱不相同。     

Antimicrobial resistance in Campylobacter, Salmonella, and Escherichia coli isolated from retail grain-fed veal meat from Southern Ontario, Canada

This study estimated the prevalence of Salmonella, Campylobacter, and Escherichia coli isolates in fresh retail grain-fed veal obtained in Ontario, Canada. The prevalence and antimicrobial resistance patterns were examined for points of public health significance. Veal samples (n = 528) were collected from February 2003 through May 2004. Twenty-one Salmonella isolates were recovered from 18 (4%) of 438 samples and underwent antimicrobial susceptibility testing. Resistance to one or more antimicrobials was found in 6 (29%) of 21 Salmonella isolates; 5 (24%) of 21 isolates were resistant to five or more antimicrobials. No resistance to antimicrobials of very high human health importance was observed. Ampicillin-chloramphenicolstreptomycin-sulfamethoxazole-tetracycline resistance was found in 5 (3%) of 21 Salmonella isolates. Campylobacter isolates were recovered from 5 (1%) of 438 samples; 6 isolates underwent antimicrobial susceptibility testing. Resistance to one or more antimicrobials was documented in 3 (50%) of 6 Campylobacter isolates. No Campylobacter isolates were resistant to five or more antimicrobials or category I antimicrobials. E. coli isolates were recovered from 387 (88%) of 438 samples; 1,258 isolates underwent antimicrobial susceptibility testing. Resistance to one or more antimicrobials was found in 678 (54%) of 1,258 E. coli isolates; 128 (10%) of 1,258 were resistant to five or more antimicrobials. Five (0.4%) and 7 (0.6%) of 1,258 E. coli isolates were resistant to ceftiofur and ceftriaxone, respectively, while 34 (3%) of 1,258 were resistant to nalidixic acid. Ciprofloxacin resistance was not detected. There were 101 different resistance patterns observed among E. coli isolates; resistance to tetracycline alone (12.7%, 161 of 1,258) was most frequently observed. This study provides baseline prevalence and antimicrobial resistance data and highlights potential public health concerns.     

Typing of Escherichia coli O157 strains isolated from fresh sausage

E. coli O157 is a foodborne pathogen responsible for serious human illnesses, such as hemorrhagic colitic and hemolytic uremic syndrome. Ground beef products are among the foods that are more commonly contaminated, and the strains isolated have been frequently found to carry virulence factors of this pathotype. This paper reports the results of serotyping assays and of investigations performed to screen for virulence factors of 10 E. coli O157 strains isolated from fresh sausages purchased at retail meat outlets in various provinces of Apulia (southern Italy). The presence of verocytotoxins was assessed on VERO cells and ELISA tests. Multiplex PCR assays were performed for the eae, stx₁, stx₂ and hlyA genes. Six of the 10 strains examined presented the H7 antigen and all of them proved to be potentially pathogenic due to the presence of individual or multiple virulence factors.     

Characterization of resistance patterns and detection of apramycin resistance genes in Escherichia coli isolated from swine exposed to various environmental conditions

Weaned pigs were separated into eight treatments including a control without exposure to apramycin; a control with exposure to apramycin; and apramycin plus either cold stress, heat stress, overcrowding, intermingling, poor sanitation, or intervention with oxytetracycline, to determine the effects of management and environmental conditions on antibiotic resistance among indigenous Escherichia coli. Pigs exposed to apramycin sulfate received that antibiotic in the feed at a concentration of 150 g/ton for 14 days. Environmental treatments were applied 5 days following initial antibiotic administration and maintained throughout the study. Fecal samples were obtained on day 0 (prior to antibiotic treatment) and on days 2, 7, 14, 28, 64, 148, and 149. E. coli were isolated and tested for resistance to apramycin using a minimum inhibitory concentration (MIC) broth microdilution method. Macrorestriction profiling, arbitrarily primed PCR, PCR targeting a gene coding for apramycin resistance, and DNA hybridization were used to characterize genetic elements of resistance. Increased (P<0.0001) resistance to apramycin was noted in E. coli from all treatment groups administered apramycin. MICs of isolates from control pigs receiving apramycin returned to pretreatment levels following removal of the antibiotic, whereas isolates from cold stress, overcrowding, and oxytetracycline groups expressed greater (P<0.05) MICs through day 64, before returning to pretreatment levels. Genetic analysis indicated that all resistant isolates carried the aac(3)IV gene sequence and this sequence was found in a variety of E. coli isotypes. Our data indicate that E. coli resistance to apramycin is increased upon exposure to various stressors.     

Antimicrobial resistance in Campylobacter coli isolated from pigs in two provinces of China

The aim of this study was to determine the prevalence, antimicrobial resistance and molecular epidemiology of Campylobacter coli isolated from swine in China. A total of 190 C. coli isolates obtained from two slaughter houses and ten conventional pig farms in Shandong (SD, n = 95) and Ningxia (NX, n = 95) provinces were tested for their susceptibility to 14 antimicrobials. A high prevalence (> 95%) of ciprofloxacin and tetracycline-resistant strains was observed in both SD and NX. The erythromycin and clindamycin resistance rates of C. coli from NX (ERY: 54.7% CLI: 43.2%) were higher than those from SD (ERY: 37.9%, CLI: 35.8%). A significant difference (P < 0.05) was observed in erythromycin resistance rate, but not (P > 0.05) in clindamycin resistance rate. while the resistance rates of ampicillin and kanamycin in NX (AMP: 34.7%, KAN: 43.2%) were significantly lower (P < 0.05) than those in SD (AMP: 51.6%, KAN: 71.6%). None of the tested isolates were resistant to phenicols. The majority of the isolates from both provinces (SD: 80% and NX: 73.7%) showed multi-drug resistance profiles. The point mutations of A2075G in the 23S rRNA and C257T in the gyrA gene were detected in 98% (87/89) of macrolide resistant isolates and all ciprofloxacin resistant isolates, respectively. In addition, all tetracycline-resistant isolates harbored the tet(O) gene. The high prevalence of antimicrobial resistance in C. coli strains derived from pigs in China was observed and was likely due to the extensive use of various antimicrobials. Prudent use of antimicrobial agents on farms should be further emphasized to control the dissemination of antimicrobial resistant C. coli.     

养猪场源致泻大肠埃希菌毒力基因和致病型研究

目的 了解养猪场源致泻大肠埃希菌(DEC)毒力基因及致病型分布情况,为从猪肉生产源头防控该菌引发的食源性疾病提供参考数据.方法 采用多重荧光实时定量聚合酶链式反应(PCR)检测分离 自12个养猪场的生猪、养殖环境和养殖工人的大肠埃希菌毒力基因,鉴定DEC的致病型.结果 985株分离 自养猪场的大肠埃希菌中,DEC占比2...     

Antimicrobial properties of lactic acid bacteria and yeast-LAB cultures isolated from traditional fermented milk against pathogenic Escherichia coli and Salmonella enteritidis strains

The survival and growth of Escherichia coli 3339 and Salmonella enteritidis 949575 isolated from human clinical samples, in milk fermented with lactic acid bacteria (LAB) and yeast strains previously isolated from Zimbabwean naturally fermented milk (NFM) was studied. The LAB starter cultures used were Lactococcus lactis subsp. lactis biovar. diacetylactis C1 alone (C1) or in combination with Candida kefyr 23 (C1/23), L. lactis subsp. lactis Lc261 alone (LC261) or in combination with C. kefyr 23 (Lc261/23). The growth of the same pathogens in milk fermented with a commercial DL culture (CH-N 22) and spontaneously fermented raw milk was also monitored. The C1 and C1/23 cultures significantly (P<0.05) inhibited the growth of both pathogens. When inoculated at the beginning of the fermentation, both E. coli 3339 and S. enteritidis 949575 counts were significantly (P<0.05) reduced by about two log cycles in C1 and C1/23 cultured milk. However, in naturally fermented milk and the DL cultured milk, both E. coli 3339 and S. enteritidis 949575 grew and reached high populations of about 9 and 8.8 log cfu ml(-1), respectively, after 18 h. When E. coli 3339 was inoculated into previously fermented milk, the viable counts were significantly (P<0.05) reduced in the presence of C1 and C1/23 from 7 log cfu ml(-1) to 3 log cfu ml(-1) after 48 h. S. enteritidis 949575 could not be recovered from these cultures after 48 h. The addition of the yeast did not enhance or diminish the inhibitory capacity of the LAB cultures. The pathogens survived in high numbers when inoculated into pre-fermented NFM and the commercial DL- (CH-N 22) cultured milk. The C1 strain, therefore, offered the best protection against the pathogens. Its inhibitory effect was mainly related to fast acid production.     

成都市海产品中副溶血性弧菌耐药特征及多位点序列分型

目的了解成都市不同种类的海产品中副溶血性弧菌的污染程度、耐药情况、毒力基因分布、基因分型情况,为成都市食源性副溶血性弧菌流行及其风险评估提供基础数据。方法参照GB 4789. 7—2013《食品安全国家标准食品微生物学检验副溶血性弧菌检验》,从不同种类的海产品中分离副溶血性弧菌疑似菌株,通过生化试验及16S r DNA测序进行准确鉴定;采用纸片扩散法对分离株进行药敏试验,通过聚合酶链式反应(PCR)检测与其致病性相关的2个毒力基因,对分离株进行多位点序列分型分析。结果从采集的380份海产品中共104份样品检出副溶血性弧菌,总检出率为27. 4%。药敏试验表明,97. 1%(101/104)的分离株具有耐药性,其中对氨苄西林的耐药率最高(95. 2%,99/104)。分离株trh基因携带率为12. 5%(13/104),tdh基因携带率为1. 0%(1/104); 104株分离株共分为38个ST型,其中ST1801、ST392、ST413型分离率较高,分离株未出现流行克隆群。结论流通过程中不同种类海产品副溶血性弧菌污染率、耐药情况、毒力基因分布存在差异,可能与养殖环境、运输条件等有关。