Antimicrobial resistance of Escherichia coli isolated from chickens

Faecal Escherichia coli isolated from healthy farm chickens, from farm chickens with avian influenza, and from chickens with diarrhoea were more resistant to antimicrobial agents (94-100%) than those isolated from healthy domestic chickens (20%). Transfer of drug resistance was readily achieved from strains isolated from both healthy and sick farm chickens, and from diarrhoeic chickens; it was more difficult to demonstrate in strains from domestic chickens. Resistant E. coli showing serotypes suspected to be enteropathogenic for man, i.e 0126:K71(B16), 044:K74 (L) and 0119:K69(B14), were isolated from faecal samples of healthy and sick farm chickens, but not from healthy domestic birds.     

鸡源耐安普霉素大肠杆菌耐药表型研究

进行了安普霉素耐药大肠杆菌耐药表型的研究.采用常规方法和生化鉴定管对有过安普霉素用药史的鸡场分离的鸡源病原性大肠杆菌进行鉴定,并用试管二倍稀释法测定其最低抑菌浓度(MIC),筛选出鸡源安普霉素耐药大肠杆菌;采用药敏纸片法研究了这些耐药菌对安普霉素等14种抗菌药物的敏感性.共筛选出7株对安普霉素耐药的鸡源性大肠杆菌,这些耐药菌株全部对安普霉素、妥布霉素、奈啶酸、多西环素和阿莫西林耐药;大部分对庆大霉素、链霉素、卡那霉素、壮观霉素也呈现耐药.对新霉素的耐药较低,对阿米卡星高度敏感.部分交叉耐药现象的存在揭示对安普霉素等氨基糖苷类药物产生耐药性的菌株,其他抗生素也可能对它们失去疗效.     

鸡致病性大肠杆菌对头孢噻呋和氟喹诺酮类药物的敏感性

近年来,第三代动物专用头孢菌素类头孢噻呋和发展极为迅速的氟喹诺酮类药物均在兽医临床上广泛应用,取得了明显的治疗效果。但随着使用时间的延长和用药的增多,临床上细菌耐药性也开始出现并导致疗效下降。然而,目前尚缺乏系统资料说明其耐药性发生的程度。为了更好地控制鸡大肠杆菌病,本研究对2004-2005年河南省中北部9个地级市区大型养殖场收集的91株鸡源致病性大肠杆菌对头孢噻呋和氟喹诺酮类药物的耐药性情况进行了观查。1材料与方法1.1药品与试剂头孢噻呋、恩诺沙星、环丙沙星和沙拉沙星标准品均购自中国兽医药品监察所;细菌生化鉴定管…     

广东鸡致病性大肠杆菌对氟喹诺酮类药物的敏感性分析

以广东省21个地级市为单位,从鸡场病死鸡分离细菌,选取经生化和动物试验证明的鸡致病性大肠杆菌共115株,采用药敏纸片法测定其对氟喹诺酮类药物的敏感性.结果显示,72.2 %的试验菌对氟喹诺酮类药物产生抗药性;氟喹诺酮类药物之间交叉耐药严重,恩诺沙星与氟罗沙星交叉耐药率高达99%.     

Molecular characteristics of ESBL-producing Escherichia coli isolated from chickens with colibacillosis

BackgroundAvian pathogenic Escherichia coli (APEC) causes colibacillosis, resulting in significant economic losses in the poultry industry.ObjectivesIn this study, the molecular characteristics of two extended-spectrum beta-lactamase (ESBL)-producing APEC isolates were compared with previously reported ESBL-producing E. coli isolates.MethodsThe molecular characteristics of E. coli isolates and the genetic environments of the ESBL genes were investigated using whole genome sequencing.ResultsThe two ESBL-producing APEC were classified into the phylogenetic groups C and B1 and ST410 and ST162, respectively. Moreover, the ESBL genes of the two isolates were harbored in different Inc plasmids. The EC1809182 strain, harboring the bla_CTX-M-55 gene on the plasmid, exhibited extensive homology to IncFIB (98.4%) and IncFIC(FII) (95.8%). The EC1809191 strain, harboring the bla_CTX-M-1 gene, was homologous to IncI1-I (Gamma) (99.3%). All chromosomes carried the multidrug transporter, mdf(A) gene. Mobile genetic elements, adjacent to CTX-M genes, facilitated the dissemination of genes in the two isolates, analogous to other ESBL-producing E. coli isolates.ConclusionsThis study clarifies the transmission dynamics of CTX-M genes and supports strengthened surveillance to prevent the transmission of the antimicrobial-resistant genes to humans via the food chain.     

Cytotoxin produced by Escherichia coli isolated from chickens with swollen head syndrome (SHS)

This study was conducted to determine the presence of selected virulence factors in Escherichia coli recovered from swollen head syndrome (SHS) in chickens. Thirty-six (72%) of 50 isolates of E. coli from SHS were shown to produce a cytotoxin that was active on Vero and HeLa but not on Y-1 cells. The toxin was called VT2y because the cytotoxic effect on Vero cells was morphologically similar to that produced by E. coli verotoxins (VTs), and this effect was neutralized by antiserum against VT2 but not by antiserum against VT1. Interestingly, DNA probes for VT1 and VT2 failed to hybridize the VT2y-positive isolates with total DNA. Although the role of this toxin in SHS has yet to be demonstrated, the marked edema which characterizes affected chickens is consistent with the action of a toxin such as verotoxin which targets vascular endothelium.     

Congo red binding by Escherichia coli isolates from chickens

An attempt was made to use a recently reported special Congo red medium to determine the pathogenicity of Escherichia coli isolates obtained from chickens. The inclusion of bile salts in the Congo red medium as described in previous reports by others was found in the current experiments to cause the production of red colonies from almost all E. coli cultures tested, including known Congo red-negative control cultures. Cultures of E. coli, regardless of their pathogenic history, rarely produced red colonies on the Congo red medium without added bile salts. Numerous isolates of other bacterial genera were examined and found to produce red colonies on the Congo red medium with or without added bile salts.     

Antibiotic resistance of Escherichia coli strains isolated from chickens with colisepticaemia in Morocco

Sixty-two strains of Escherichia coli were isolated in 58 farms from broiler chickens showing respiratory signs and lesions characteristic of avian colibacillosis. Serological examination of these strains showed that the types 078, 01 and 02 (for the somatic antigen) and K1 (for the capsular antigen) were the most frequently found. Newcastle disease virus was also isolated in two cases. All the strains of E. coli isolated were sensitive to colistin, flumequine and gentamicin. A few strains were resistant to neomycin, nalidixic acid and trimethoprim. The frequency of strains resistant to nitrofurans, sulfonamides, chloramphenicol, spectinomycin and ampicillin was intermediate. Most strains were resistant to tetracycline. Multiple resistance was common.     

Spatial heterogeneity of Escherichia coli DNA fingerprints isolated from cellulitis lesions in chickens

Avian cellulitis in broiler chickens is characterized by subcutaneous lesions that result in economic losses because of the partial or complete condemnation of the carcasses at processing. Escherichia coli is the primary causative agent of this condition. Previous research with a biotyping system found that the E. coli of cellulitis origin were unique to each ranch, suggesting that these E. coli were endemic within the ranch environment. The objective of our study was to analyze the genetic variability of E. coli isolates associated with cellulitis. We analyzed the genetic relatedness of the isolates in relation to the houses, ranches, and complexes in which the broilers were grown. This analysis enabled us to assess the spatial heterogeneity, or genetic diversity on a spatial scale, of the isolates. Forty-nine broilers with cellulitis lesions were necropsied. These broilers came from six houses on four ranches on three complexes that had been placed with chicks from the same hatchery within a 2-wk period. Isolates of E. coli from the lesions were DNA fingerprinted by pulsed-field gel electrophoresis. Relatedness among isolates was determined with the Dice coefficient and an unweighted pair group method with average linkages cluster analysis. The complexes possessed isolates with a variety of DNA fingerprints, yet each complex appeared to have isolates with a unique set of DNA fingerprints. Isolates from the same complex tended to form clusters with similarity coefficients greater than 90%. Isolates from different complexes were genetically distinct. This heterogeneity at the level of the complex suggests that isolates were not disseminated from a source common to the complexes. The spatial heterogeneity of the E. coli isolates in this study implies an endemic population of cellulitis-associated E. coli exists in the broiler house environment.     

Serotypes of Escherichia coli isolated from septicaemic chickens in Galicia (Northwest Spain)

The purpose of this study was to establish the serogroups of Escherichia coli that cause avian colibacillosis in Spain. The serogroups of 625 avian E. coli isolated between 1992 and 1993 were determined. The 458 E. coli from chickens with septicaemia belonged to 62 different O serogroups; however, 59% were of one of 18 serogroups (O1, O2, O5, O8, O12, O14, O15, O18, O20, O53, O78, O81, O83, O102, O103, O115, O116 and O132). These 18 serogroups were also determined as an important percentage (29%) of control isolates from faeces of healthy birds. Nevertheless, a significant difference (59% versus 29%; P<0.001) was observed. Furthermore, the serogroups O12, O14, O18, O53, O78, O81, O102, O115, O116 and O132 were almost exclusively identified among septicaemic E. coli (31% versus 3%; P<0.001). The high prevalence of O18, O81, O115, O116 and O132 isolates was not expected and may indicate the emergence of five new serogroups associated with avian colibacillosis not yet reported.     

信宜地区鸡大肠杆菌耐药性的调查

近几年来,由致病性大肠杆菌引起的鸡大肠杆菌病在广东省信宜市有明显上升趋势,由于多方面原因,特别是临床上随意及超量使用抗菌药物,致使大肠杆菌的耐药性不断加强,呈现难以控制的局面.     

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.     

Serotyping and virulence genes detection in Escherichia coli isolated from fertile and infertile eggs, dead-in-shell embryos, and chickens with yolk sac infection

Escherichia coli is a common avian pathogen mainly associated with extraintestinal infections such as yolk sac infection (YSI). The aim of this study was to determine the serotypes and the presence of some virulence genes of E. coli strains isolated from different samples in a vertically integrated poultry operation in Mexico. Two hundred sixty-seven E. coli isolates from different samples were serotyped using rabbit serum against the 175 somatic (O) and 56 flagellar (H) antigens of the typing schema. Virulence genes were determined by colony blot hybridization, using DNA probes for st, eae, agg1, agg2, bfp, lt, cdt, slt, and ipaH diarrhea-associated virulence factors. The serogroup of 85% of the strains was determined; O19 (12%), 084 (9%), 08 (6%), and 078 (5%) were the most common. Using the complete antigenic formula (O and H), O19:NM (n = 31) was the serotype most frequently isolated from dead-in-shell embryos and in broilers that had died on the fourth, fifth, sixth, and seventh days after hatch. One hundred ten strains (41.2%) hybridized with one or more of the used probes. Of these, ipaH (72%), eae (30%), and cdt (27%) were the most common. Considering the origin of the respective isolates, 40% of the broiler farm strains were positive for at least one probe. Results show that some avian E. coli strains isolated in Mexico are included in avian pathogenic E. coli serotypes not previously reported, suggesting that they could be specific for this geographic area. The wide distribution of the ipaH gene among nonmotile strains suggests that this invasiveness trait could be important in YSI pathogenesis. On the other hand, some other genes could contribute to E. coli virulence during YSI.     

Virulence factors of avian pathogenic Escherichia coli strains isolated from chickens with colisepticemia in Japan

The virulence factors of avian pathogenic Escherichia coli (APEC) isolated in Japan were investigated. Serogroups O, serotypes K1 and K5, and genes cva C, iss, iutA, papA, tsh, and usp, which have been thought to be related to virulence, were examined for their association with E. coli strains isolated from diseased and healthy chickens. The frequently recognized serogroups O1, O2, and O78 were found in 56 of 125 (44.8%) strains of diseased chickens (APEC) versus 13 of 100 (13.0%) strains of healthy chickens (commensal E. coli), a significant difference at risk ratio < 0.01. Although iss, iutA, and tsh were widely distributed in the APEC irrespective of O serogroup, papA, usp, and the K1 serotype were detected in serogroup O2 of APEC. The kfiD gene related to the K5 capsule and VT, LT, and ST genes related to exotoxins were not detected in any strains examined.     

Serotyping of O and pilus antigens of Escherichia coli strains isolated from chickens with coli-septicemia.

One hundred and fifty one Escherichia coli strains were isolated from broiler chickens with coli-septicemia in Aichi (63 strains), Shizuoka (58 strains), and Kagoshima (30 strains) prefectures from 1980 to 1987, and their O and pilus antigens were serologically typed. One hundred and twenty five strains (82.8%) were typed into 23 O serogroups, and twenty six strains (17.2%) remained untypable. The predominant O serogroups were O2 (35 strains, 23.2%) and O78 (24 strains, 15.9%). Distribution of O serogroup was different, depending on prefectures where they are isolated. In total, 109 strains (72.2%) possessed Type 1 and/or Fmsha pili (Type 1; 41 strains, Fmsha; 22 strains, and Type 1 and Fmsha; 46 strains), and 42 strains (27.2%) were non-piliated. All the strains lacked K88, K99, 987P, F41, and Att25 pili. The ratios of piliated strains to non-piliated ones were almost the same among the three prefectures. Strains possessing Type 1 pili showed variety of O antigens, but most of the strains with Fmsha pili belonged to O2 serogroup.     

Curli expression by Escherichia coli strains isolated from bovine mastitis

The aim of the study was to determine the expression of mannose-sensitive and mannose-resistant adhesins by agglutination of cattle, sheep, goat, rabbit, horse, and chicken red blood cell assay, and curli fimbriae by Congo red binding assay among 341 E. coli strains isolated from 51 milk samples of clinically recognized bovine mastitis. Curli fimbriae expression within biofilms created on an inert surface was also investigated. To determine whether curli fimbriae are expressed both in conditions optimal for their production and in conditions resembling the host organism, the study was conducted in anaerobic atmosphere at 37 degrees C, and at room temperature in aerobic atmosphere. The results demonstrated that although the E. coli isolates examined were deprived of mannose-sensitive and mannose-resistant adhesins they were able to produce curli fimbriae in both aerobic and anaerobic conditions at room and higher temperature, indicating that these adhesins may be involved in the pathogenesis of bovine mastitis.     

Mutant-prevention concentrations of enrofloxacin for Escherichia coli isolates from chickens

OBJECTIVE: To investigate the development of enrofloxacin resistance among Escherichia coli isolates obtained from chickens by determining mutant-prevention concentrations (MPCs) and sequence the quinolone resistance-determining regions (QRDRs) of gyrA and parC genes in selected isolates. SAMPLE POPULATION: 15 chicken-derived E coli isolates. PROCEDURES: For all isolates, MPC and minimal inhibition concentration (MIC) of enrofloxacin were determined. The MPCs and maximum serum drug concentrations attained with enrofloxacin doses recommended for treatment of E coli infections in chickens were compared. Mutation frequencies and QRDR sequence changes in gyrA and parC were also determined. RESULTS: In 2 of 15 E coli strains, MPCs were low (0.016 and 0.062 microg/mL), MPC:MIC ratios were 2 and 4, and the GyrA and ParC proteins had no mutations. In 9 susceptible isolates with a GyrA point mutation, MPCs ranged from 2 to 16 microg/mL. For isolates with double mutations in GyrA and a single mutation in ParC, MPCs were > 32 microg/mL (several fold greater than the maximal plasma concentration of enrofloxacin in chickens); mutation frequencies were also much lower, compared with frequencies for single-mutation isolates. CONCLUSIONS AND CLINICAL RELEVANCE: For E coli infections of chickens, MPC appears to be useful for determining enrofloxacin-dosing strategies. The high MPC:MIC ratio may result in enrofloxacin-treatment failure in chickens infected with some wild-type gyrA E coli isolates despite the isolates' enrofloxacin susceptibility (MICs 0.125 to 1 microg/mL). For infections involving isolates with high MPCs, especially those containing mutations in gyrA and parC genes, treatment with combinations of antimicrobials should be adopted.     

Characterization of Escherichia coli isolated from healthy dogs

Five month old dogs from a Midwestern research kennel occasionally developed bloody diarrhea after shipment to other facilities. As previous diagnostic efforts failed to reveal any potential pathogens in feces from normal and diarrheic dogs, Escherichia coli was investigated for select virulence properties that may contribute to the occurrence of bloody diarrhea. Fecal swabs from 52 healthy dogs were examined for E. coli. Two hundred and sixty E. coli-like colonies were screened by PCR for the attaching and effacing (eae) gene, Shiga toxin (stx) genes, and the heat-stable enterotoxin type A (sta) gene. One hundred forty two of the 260 E. coli-like colonies (54.6%) from 43 dogs were eae or sta positive; and 60 of the eae and/or sta positive isolates were examined further. Among the 60 isolates, 23 (38.3%) possessed the eae gene, 32 (53.3%) possessed the sta gene, and five (8.3%) possessed both eae and sta genes (eae⁺/sta⁺). Of the 60 isolates, six sta⁺ and one eae⁺/sta⁺ isolates were hemolytic. When examined in the suckling mouse assay, five of six sta⁺ isolates and three of four eae⁺/sta⁺ isolates gave gut-to-remaining carcass ratios ≥0.083, indicating expression of heat-stable enterotoxin. These enterotoxin-producing isolates belonged to serogroups O42, O170, and O-negative.