Antibiotic susceptibility and molecular characterization of resistance genes among Escherichia coli and among Salmonella subsp. in chicken food chains

Objective: To investigate the occurrence of resistance genes among Escherichia coli(E. coli) and Salmonella subsp. isolated in chicken food chains in Phnom Penh, 2012–2013.Methods: Six hundred eighty two E. coli and 181 Salmonella Albany, Corvallis, and Kentucky strains were examined for susceptibilities to eight antimicrobials and following resistance genes were identified by PCR: blaTem, Str A, aad A, sul1, sul2, gyr A, Tet(A), and Tet(B).Results: E. coli presented high resistances to tetracycline, amoxicillin, and sulfamethoxazole(63.1%–76.1%). Salmonella Albany and Salmonella Kentucky traduced high resistance percentages to amoxicillin, tetracycline, sulfamethoxazole, and nalidixic acid(84.6%–100%). Among amoxicillin-resistant isolates, blaTemgenes were observed for 62% of E. coli isolates and 20% of 65 Salmonella Kentucky. The Str A gene was prevalent in 36% of 331 aminoglycoside-resistant E. coli and 90% of 40 aminoglycoside-resistant Salmonella Corvallis. The sul2 gene was predominant among sulfamethoxazole-resistant isolates, for 56% of 431 E. coli and 53% of 66 Salmonella Corvallis; the sul1 gene was observed in 54% of Salmonella Albany. The Tet(A) resistance gene was prevalent in E.coli(86%), Salmonella Corvallis(82%), Salmonella Kentucky(84%). High percentages of gyr A genes observed among nalidixic-acid resistant E. coli(91%), Salmonella Albany(92%), Salmonella Corvallis(75%) and Salmonella Kentucky(85%).Conclusions: Important occurrences of resistance gene were observed among E. coli and Salmonella in chicken food chains in Cambodia.

Detection of Drug Resistance in Escherichia coli Isolated from Animals and ERIC Analysis of Multidrug-resistant Strains

The drug resistance of Escherichia coli from animals was detected in Shandong Province in 2016,the gene mapping of Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) of multidrug-resistant isolates were analyzed,and then the relationship between ERIC-PCR genotyping and drug resistance of highly-resistant E.coli with multidrug resistance was discussed.A total of 110 E.coli isolates were separated and identified from diseased swine and avian,and their resistance to 10 kinds of antimicrobials was determined by the agar dilution method.Twenty highly-resistant isolates with multidrug resistance were selected to carry on the ERIC-PCR,followed by cluster genetic analysis according to the DNA fingerprints.The swine-sourced E.coli isolates possessed serious resistance against several kinds of antimicrobial agents,for example,all isolates were tolerant to florfenicol,doxycycline and ampicillin (100%),but had a relative lower resistance rate to cefotaxime sodium (57.14%).The same situation was observed in the poultry-sourced E.coli isolates,which had a resistance rate of 95.51% against florfenicol,while the lowest rate of 61.80% appeared on ciprofloxacin.Analysis of ERTIC showed dispersive fingerprint patterns of highly-resistant and multidrug- resistant isolates which represented a multiple clone resources,and there were certain correlation between the B genotype and the drug-resistant characteristics.It indicated that the animal-sourced E.coli isolates had a high level of drug resistance and were multidrug-resistant,which meant there was severe antibiotic resistance against not only different kinds of antibiotics but also different drugs of the same kind.The highly-resistant E.coli isolates with multidrug resistance had no apparent species preference,while their spread and pervasion posed a potential threat to the development of animal husbandry and the public health security.

Comparison of extended spectrum β-lactamasesproducing Escherichia coli with non-ESBLsproducing E.coli:drug-resistance and virulence

The virulent factors of Escherichia coil （E.cofi） play an important role in the process of pathopoiesis. The study aimed to compare drug-resistant genes and virulence genes between extended spectrum β-1actamases （ESBLs）-producing E.coli and non-ESBLs-producing E.cofi to provide a reference for physicians in management of hospital infection. From October 2010 to August 2011,96 drug-resistant strains of E. coli isolated were collected from the specimens in Qingdao Municipal Hospital, Qingdao, China. These bacteria strains were divided into a ESBLs-producing group and a non-ESBLs-producing group. Drug sensitivity tests were performed using the Kirby-Bauer （K-B） method. Disinfectant gene, qacEAl-sull and 8 virulence genes （CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1） were tested by polymerase chain reaction （PCR）. Among the 96 E.coli isolates, the ESBLs-producing E.coli comprised 46 （47.9%） strains and the non-ESBLs-producing E.cofi consisted of 50 （52.1%） strains. The detection rates of multiple drug-resistant strain, qacEAl-sull, CNF2, hlyA, eaeA,VT1, est, bfpA, elt, and CNF1 in 46 ESBLs-producing E.coli isolates were 89.1%, 76.1%, 6.5%, 69.6%, 69.6%, 89.1%, 10.9%, 26.1%, 8.7%, and 19.6%, respectively. In the non-ESBLs-producing E.cofi strains, the positive rates of multiple drug-resistant strain, qacEAl-sull, CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1 were 62.0%, 80.0%, 16.0%, 28.0%, 64.0%, 38.0%, 6.0%, 34.0%, 10.0%, and 24.0%, respectively. The difference in the detection rates of multiple drug-resistant strain, hlyA and VT1 between the ESBLs-producing E.cofi strains and the non-ESBLs-producing E.cofi strains was statistically significant （P〈0.05）. The positive rate of multiple drug-resistant strains is higher in the ESBLs-producing strains than in the non-ESBLs-producing strains. The expression of some virulence genes hlyA and VT1 varies between the ESBLs-producing strains and the non-ESBLs-producing strains. Increased awareness of clinicians and enhanced testing by laboratories are required to reduce treatment failures and prevent the spread of multiple drug-resistant strains.

Antimicrobial Drug Resistance Pattern of <i>Escherichia coli</i>Isolated from Chickens Farms with Colibacillosis Infection

Colibacillosis refers to any localized or systemic infection caused entirely or partly by avian pathogenic Escherichia coli (APEC). Colibacillosis in mammals is most often a primary enteric or urinary tract disease, whereas colibacillosis in poultry is typically a localized or systemic disease occurring secondarily when host defenses have been impaired or overwhelmed by virulent E. coli strains. The purpose of this study was to investigate the antimicrobial drug resistance pattern of Escherichia coli isolated from broiler chickens farms with colibacillosis infection. Dead birds from commercial broiler chicken farms showing signs of colibacillosis were necropsied and swab samples were collected from internal organs and blood aseptically for the isolation of Escherichia coli. Pure colonies of the bacteria were isolated on solid media and the isolates were identified as E. coli based on morphological and biochemical characteristics. For determination of susceptibility to antibacterial agents, the disc diffusion method on Muller-Hinton agar was used. The following antimicrobial agents were tested: gentamycin, oxytetracyline, colistin, ciprofloxacin, doxycycline, nalidixic acid, co-trimoxazole (trimethoprim-sulfamethoxazole), norefloxacin, lincospectin and cefuroxime. The drug resistance patterns of the organisms were determined as a percentage and reported at three levels: susceptible, intermediate and resistant. All the isolates of Escherichia coli showed resistance to several antibiotics and a pattern of multiple drug resistance was observed. The highest rate of resistance was observed against nalidixic acid (100%) and the least rate of resistance was observed against gentamycin (17%). According to the results of this research care must be taken to avoid secondary infection (colibacillosis) in chicken farms and also avoid in careless antimicrobial consumption in food animals including chickens.

Multi-Drug Resistance Pattern of Lactose Non-Fermenting <i>Escherichia coli</i>as Causative Agent of Urine Tract Infections in Luanda, Angola

This prospective study was carried out to assess the sensitivity and resistance pattern of lactose non-fermenting Escherichia coli from July 2018 to December 2018 in the Laboratory of Microbiology at Luanda Medical Center, Angola. Out of 1170 patient, a total of 120 urine specimens infected with Escherichia coli (>105 CFU/ml) were collected according to the routine protocol of urinalysis. Among these 120 isolates, 25 (21%) isolates were determined as “atypical”, lactose non-fermenting E. colis trains. The twenty-five lactose non-fermenting Escherichia coli strains isolated from urine samples in Luanda Medical Center were declared as Multiple Drugs-Resistant strains with high resistance to Cefalexine (100%), Cefuroxime (100%), Ceftriaxone (92%), Gentamycin (92%), Ciprofloxacin (72%) and Amoxiciclin/Clavulanic (80%). The alarming resistance level to the first-choice drugs for the treatment of urinary tract infections caused by non-fermentative lactose E. coli was observed.

Monitoring and Analysis on Multi Drug Resistance of Escherichia coli from Captive Population Amur Tiger

In order to investigate the multi drug resistance to Escherichia coli from captive population Amur tiger,E. coli strains were isolated from the fecal samples of tiger in Heilongjiang Amur Tiger Park in Harbin. The sensitivity of E. coli isolates to 14 antibiotics was determined by scrip diffusion method. The results indicated that all the isolates varied in drug resistance to different antibiotics; the isolates gave high resistance to ampicillin,with a drug fast rate of 100%; over80% of the isolates were resistant to tetracycline and Paediatric Compound Sulfamethoxazole Tablets(SMZ- TMP),and over 70% of the isolates were sensitive to aztreonam,amoxicillin /potassium clavulanate. Most of the isolates had high sensitive to aztreonam and amoxicillin / clavulanate acid.

Antibiotic Sensitivity Test and Detection of Sulfonamide Resistance Gene Sul2 in Escherichia coli Isolates from Swine

[ Objective] This study aimed to analyze antibiotic sensitivity and detect sulfonamide resistance gene Sul2 in 17 Escherichia coli isolates from swine. [ Method] The antibiotic sensitivity of 17 E. coil isolates from swine was analyzed with Kirby-Bauer disk diffusion method. Sulfonamide resistance gene Sul2 was detected by PCR amplification with the extracted genomic DNA as a template. [ Result] The 17 swine-derived E. coli isolates from different regions were resistant to various antibiotics and exhibited different multi-antibiotic resistance phenotypes, including nine isolates resistant to sulfamethoxazole, seven isolates resistant to oxacillin, ten isolates resistant to cefazolin and eight isolates resistant to erythromycin. Sulfonamide resistance gene Sul2 was successfully amplified from genomic DNA of E. coli isolates. The PCR results were basically consistent with antibiotic resistance phenotypes of these strains. [ Conclusion] Sul2 gene was widespread in swine-derived E. coli, which was closely associated with sulfonamide resistance phenotvpes of E. coli.

Study on the Resistance of Pathogenic Escherichia coli to Ceftiofur

[Objective] Ceftiofur was as the substrate to induce the standard strain of Escherichia coli（E.coli）to be the drug-resistance one.The resistant mechanism of E.coli to ceftiofur was studied.[Method] The sub-inhibitory concentration method was used to induce the standard strains C83907 and C83845.After they were induced for 10 generations,the double disc synergy test（DDST）,NCCLS（National Committee for Clinical Laboratory Standards）confirmatory test and PCR amplification were used to detect the extend spectrum β-lactamases（ESBLs）.The two fold dilution method was used to measure the minimal inhibitory concentration（MIC）of cetiofur to the strain which produced ESBLs.For the drug-resistance strain which produced ESBLs,the two fold dilution method was used to measure the minimal inhibitory concentrations of different proportions of cetiofur and tazobactam sodium.[Result] After they were induced 15 generations,MIC value of ceftiofur to the induced bacteria was during 8-10 μg/ml,and ESBLs was detected.MICs of cetiofur combining tazobactam sodium（the mass ratio was 1∶1-8∶1）to Escherichia coli produced ESBLs reduced 20-22 times than that of cetiofur.[Conclusion] The main mechanism of pathogenic Escherichia coli resistance to ceftiofur was that which produced ESBLs.

Use of a naturally occurring codon bias for identifying topoisomerase mutations in ciprofloxacin resistant <i>Escherichia coli</i>using PCR and future prospects with other bacterial genera: A pilot study

We developed a novel PCR method aimed at identi- fying and amplifying native codon sequences of muta- tion-prone amino acids in DNA gyrase implicated in quinolone resistance using a naturally occurring co- don bias in E. coli DNA gyrase A.

Isolation and Identification of Drug-resistant Escherichia coli Strains from Chickens

[Objective] This study aimed to isolate and identify the drug-resistant Es- cherichia coli strains from chickens. [Method] E. coli strains were isolated from the fecal samples collected from five chicken farms around Shangqiu City, and verified by biochemical and pathogenic assay. [Result] Among the 35 isolated E. coli stains, 11 E. coil stains were sensitive to florfenicol, amikacin, neomycin and gentamicin; 12 E. coli stains were moderately sensitive to ciprofloxacin, doxycycline and norfloxacin; 15 E. coil stains were resistant against erythromycin, penicillin and streptomycin. [Conclusion] Strengthening biosecurity measures, rationally using vaccine and choosing effective antibiotics are the most cost-efficient methods to control E. coli.

Analysis of the Drug Sensitivity of Escherichia coli Isolated from Sheep in Yangling,Shaanxi Province

This study was conducted to investigate the drug sensitivity of Escherichia coli isolated from sheep, providing data reference for clinical medication. Fifty-four samples were collected from a certain sheep farm in Yangling, Shaanxi Province for E. coli isolation and identification, and K-B disk diffusion method was used to carry out the dug sensitivity test of 40 E. coli to the eight antimicrobials of spectinomycin, amikacin, tobramycin, enrofloxacin, nalidixic acid, levofloxacin, salafloxacin and ciprofloxacin. The results showed that there were 40 positive E. coli strains and the isolation rate was 74.7%. The resistance rates of the 40 E. coli to spectinomycin, amikacin,tobramycin and nalidixic acid were 85%, 95%, 97.5% and 87.5%, respectively, while the resistance rate to enrofloxacin, levofloxacin, salafloxacin and ciprofloxacin were relatively low and the resistance rates were 35%, 20%, 25% and 17.5%, respectively. The isolates showed different levels of drug resistance to eight antimicrobials and the highest resistance rate was 97.5%. It reminds that the sheep farm should strictly control the dosage according to the antimicrobial sensitive data in clinic.

Prevalence,serotyping and drug susceptibility patterns of Escherichia coli isolates from kidney transplanted patients with urinary tract infections

BACKGROUND Extended-spectrumβ-lactamase(ESBL)-producing Escherichia coli(E.coli)are among the main pathogens in urinary tract infections(UTIs)among kidney transplant patients(KTPs).AIM To estimate the prevalence of ESBL-producing E.coli in KTPs and to evaluate the most prevalent serotypes and antibacterial susceptibility patterns of isolated bacteria in Tehran,Iran.METHODS A total of 60 clinical isolates of uropathogenic E.coli were collected from 3 kidney transplant centers from April to May 2019.Antimicrobial susceptibility testing was performed by the disk diffusion method as recommended by the Clinical Laboratory and Standards Institute.The serotyping of E.coli isolates was performed by the slide agglutination method.The presence of blaTEM,blaSHV,and bla CTX-M genes was evaluated by polymerase chain reaction.RESULTS The frequency of ESBL-producing E.coli in KTPs was found to be 33.4%.All of the 60 E.coli isolates were found to be susceptible to doripenem(100%)and ertapenem(100%).High resistance rates to ampicillin(86%),cefotaxime(80%),and cefazolin(77%)were also documented.The most frequent serotypes were serotype I(50%),serotype II(15%),serotype III(25%),and serotype VI(10%).The gene most frequently found was blaTEM(55%),followed by blaCTX-M(51%)and blaSHV(41%).CONCLUSION Molecular analysis showed that blaTEM was the most common ESBL-encoding gene.The high resistance toβ-lactams antibiotics(i.e.,ampicillin,cefotaxime,and cefazolin)found in E.coli from KTPs with UTIs remains a serious clinical challenge.Further efforts to control ESBL-producing E.coli should include the careful use of all antibiotics as well as barrier precautions to reduce spread.

Escherichia coli tetracycline efflux determinants in relation to tetracycline residues in chicken

Objective:To screen for Escherichia coli(E.coli)resistant to tetracycline,followed by identification of tet efflux genes by polymerase chain reaction(PCR).In addition,detection of tetracycline residues in chicken livers and kidneys were conducted using high performance liquid chromatography-tandem quadrupole mass spectrometry(HPLC-MS-MS).Methods:Strains of E.coli were isolated from samples of chicken colon and screened for tetracycline resistance.Tetracycline genes conferring resistance(Tc^r)were detected by polymerase chain reaction(PCR).Most of the isolates were resistant to tetracycline(97.9%).Results:PCR analysis indicated that Tc^r E.coli R-plasmids contained tet(A),tet(B)and a combination of both efflux genes.None of the isolates contained other efflux tet genes tet(C,D,E and Y).High performance liquid chromatography-tandem quadrupole mass spectrometry(HPLC-MS-MS),a sensitive technique,was used to detect residues of chlortetracycline(CTC),oxytetracyeline(OTC),doxveycline(DC)in chicken livers and kidneys.The samples containing tetracycline residues were at 0.13-0.65pg/μL levels.Conclusions:Tetracycline and other antibiotics are commonly used in the poultry and meat production industry for prevention of microbial infections.Multiple antibiotic resistant bacteria in Oman have increased to alarming levels,threatening public health,domestic and may have adverse effect on environment.

In vitro biofilm formation by uropathogenic Escherichia coli and their antimicrobial susceptibility pattern

Objective:To detect in vitro biofilm formation of uropathogenic Escherichia coli(E.coli)(UPEC) strains isolated from urine specimens and also to determine their antimicrobial susceptibility pattern using 13 commonly used antibiotics.Methods:The present study comprised of 166 urine specimens collected from tertiary care hospitals in and around Coimbatore.South India. All the specimens were subjected to gram staining,bacterial culture and the E.coli strains were screened for biofilm formation using Tube Method(TM),Congo Red Agar(CRA) and Tissue Culture Plate method(TCP) respectively.Subsequently,the antimicrobial susceptibility test was performed by Kirby Bauer-disk diffusion method for the biofilm and non-biofilm producing E. coli strains.Results:Of the 100(60.2%) E.coli strains,72 strains displayed a biofilm positive phenotype under the optimized conditions in the Tube Method and the strains were classified as highly positive(17,23.6%),moderate positive(19.26.3%) and weakly positive(36.50.0%). similarly under the optimized conditions on Congo Red agar medium,biofilm positive phenotype strains were classified as highly positive(23,23%).moderate positive(37.37%) and weakly positive(40,40%).While in TCP method,the biofilm positive phenotype strains were also classified as highly positive(6.6%),moderate positive(80.80%) and weakly positive(14,14%),it didn’t not correlate well with the tube method for detecting biofilm formation in E.coli.The rates of antibiotic resistance of biofilm producing E.coli were found to be 100%for chloramphenicol and amoxyclav(amoxicillin and clavulanic acid),86%for gentamicin and cefotaxime.84%for ceftazidime,83%for cotrimoxazole and piperacillin/tazobactam,75%for tetracycline and 70% for amikacin,Conclusions:This study reveals the prevalence and antimicrobial susceptibility pattern of biofilm and non-biofilm producing uropathogenic E.coli strains.

Performance Parameters:Demobilization Antibiotic Resistant Bacteria(ARB)and Carrying Genes(ARG)in Wastewater Disinfection

The UV irradiation is used for removing Antibiotic Resistant Bacteria(ARB)and Antibiotic Resistance Genes(ARG)from wastewater treatment.Bacteriophages are viruses that infect within bacteria,are recognized for bacterial control.The influence of some parameters in quantification and performance influencing of pathogen demobilization could be considered in disinfection of wastewater.The comparison of Polyvalent phage(NE1)versus Coliphage(NE4)in suppressing a bacterium Escherichia coli(NDM-1:b-lactam-resistant)with UV irradiation was observed the efficacy in reduction of cells in the disinfection and parameter process.The results with the effect of UV-C irradiation on NDM-1 infected with 1%of NE4 showed a decrease of cells from 8×10^(6)to 2×10^(5)in 60 min with UV-C dose.The NDM1(E.coli)was infected with 1%of NE4(Polyvalent Phage)under magnetic stirring for 1 h,the cells count was 8×10^(6).After 1 h in UV-C e×posure,the cells number reached 3×10^(5).The NDM1 that was e×posed in 1 h of UV-C irradiation and then was infected with 1%of NE4.Cells counting were done 24 h after this procedure.These cells were e×posed in UV-C and showed a reduction in the number of cells from 1×10^(8)to 4×10^(5)after 60 min.The results indicate that bacteriophages can mitigate bacteria species,and combined the conventional water disinfection technologies that can support the microbial safety control strategies.

Antiboitic Resistance of Uropathogenic Eshcherichia coli in Pateints of Hargeisa Group Hospital, Hargeisa, Somaliand

Background: Urinary tract infection is a common disease in Somaliland society. The predominant causative organism of Urinary tract infection is Escherichia coli. This research studies antibiotic resistance of uropathogenic E. coli in patients of Hargeisa Group Hospital. The study selected commonly prescribed antibiotics for urinary tract infection treatment. Methodology: Urine samples of patients were cultured to isolate causative organisms of the urinary tract infection. Chromo-agar media, CLED, and biochemical tests are applied to identify the type of bacteria. Antibiotic reactions to E. coli bacteria are measured to differentiate between sensitive and resistant drugs with the guidance of the Clinical and Laboratories Standard Institute (CLSI). Kirby Bauer disc diffusion method is applied to assess antimicrobial activity against E. coli. Data of patients such as age, sex, symptoms of UTI, previous UTI infection, and history of antibiotic use were recorded. SPSS and Microsoft Excel are applied to analyze and interpret data. Results: The predominant organism that caused urinary tract infection was Escherichia coli (55%), Klebsiella spp (15%), Candida spp (15%), Enterococcus spp (10%), Staph spp 2.5%, and Pseudomonas spp 2.5% while other 55% were negative. The study assessed antibiotic resistance of E. coli, which reported resistance to Tetracycline at (70%), Ampicillin (64%), and Cotrimoxazole (61%). The bacteria showed moderate resistance to Ceftriaxone (43.5%), Nalidixic acid (43%), and Ciprofloxacin (36%). The bacteria are sensitive to Amikacin (100%), Nitrofurantoin (96%), Levofloxacin (73%) and gentamicin (74%). Conclusion: The overall incidence of antibiotic resistance to E. coli is high because the bacteria show a percentage of resistance to each antibiotic except Amikacin which gives (100%) sensitivity. The research recommends public awareness of the risks associated with antibiotic use and periodic evaluation of antibiotic resistance to accomplish better managing urinary tract infections.

携带bla_(CTX-M)禽大肠杆菌的多重耐药特征和体外致病性研究

【目的】探究携带bla_(CTX-M)禽大肠杆菌的分子流行特点。【方法】检测76株携带bla_(CTX-M)禽大肠杆菌的多重耐药谱、系统进化分群及毒力基因,并分析毒力基因与耐药特点、系统进化分群之间的关联性。【结果】药敏结果显示,携带bla_(CTX-M)禽大肠杆菌对氟苯尼考的耐药率最高,达85.53%(65/76);其次是恩诺沙星,为48.68%(37/76);最常见的多重耐药谱为氟苯尼考+恩诺沙星+乙酰甲喹,达10.53%(8/76)。系统进化分群结果显示,主要的亚群为C群和B1群;其中,C群检出率最高,达53.95%(41/76);其次是B1群,为30.26%(23/76);E群、A群、D群和F群的检出率分别为9.21%(7/76)、2.63%(2/76)、2.63%(2/76)和1.32%(1/76);未检测出B2群和分支I群。毒力基因检测结果显示,76株携带bla_(CTX-M)禽大肠杆菌均检出了至少4种毒力因子,且有11株菌同时携带至少11种毒力基因;其中,有13株菌同时检出禽致病性大肠杆菌(avian pathogenic Escherichia coli,APEC)的5个标志性毒力基因(iroN、iutA、hly、OmpT和iss),检出率为17.11%;有11株菌检出ColV质粒的标志基因(cva/cvi、iroN、iucD/iutA、etsAC、OmpT/hlyF和sitA),检出率为14.47%;有28株菌检出耶尔森菌强毒力岛(high pathogenitility island,HPI)的标志基因irp-2和fyuA,检出率为36.82%。【结论】携带bla_(CTX-M)禽大肠杆菌多为多重耐药菌,且多重耐药菌株携带ColV毒力质粒可能性更高;携带bla_(CTX-M)禽大肠杆菌的多重耐药与致病性有关联,即携带bla_(CTX-M)禽大肠杆菌具有多重耐药的同时,具有致病的可能性更高。

Isolation and Characterization of E. Coli O157 : H7 from Infected Newborn Calves in Northeast China

Escherichia coli O157 : H7 is a foodborne pathogen that poses a major threat to public health. Epidemiologic investigations have identified dairy cows, especially calves, are the principal reservoir of E. coli O157 : H7. In this study, based on the results, E. coli O157 : H7 was the main cause of E. coli disease outbreak in late October, 2015, and more than 90% of newborn calves died of serious diarrhea. Through further experiments, the drug sensitivity and resistance of the strain, the expression of the virulence gene and virulence pathogenicity were studied. E. coli O157 : H7 isolates were resistant to 12 antibiotics including penicillin, tetracycline and ampicillin, and were sensitive to eight antibiotics including cefoperazone, ceftazidime and amikacin. Resistance genes included tetB, strB, aadB, aphA, floR, TEM and virulence genes included stx1, eaeA and hlyA. Using specific pathogen free mice, the result showed that the isolate was pathogenic with a median lethal dose of 7.9×107 CFU · mL-1. This study described the pathogenesis and clinical manifestations of E. coli O157 : H7 infection. These results guided the use of antibiotics in prevent and control of bacterial infections in the future.

通辽部分地区鸭源大肠杆菌耐药性检测及多重耐药菌株YA-1全基因组测序分析

【目的】大肠杆菌作为家禽最常见细菌性病原之一,其耐药性的不断增强已引起广泛关注。试验旨在了解通辽地区部分鸭场大肠杆菌的耐药性及耐药基因分布,为该地区鸭源大肠杆菌耐药性及耐药基因的研究提供参考。【方法】经16S rRNA测序、生化试验和小鼠致病性试验对送检的通辽市各旗县15份病死鸭肠道样品进行病原菌分离纯化,对分离菌株进行药敏试验及耐药基因的PCR检测,并选取多重耐药菌株进行全基因组测序。【结果】所分离的10株鸭源大肠杆菌均具有不同程度耐药性,对链霉素、环丙沙星、恩诺沙星等药物的耐药率在80%及以上,耐药基因aphA1、strB和qnrS的检出率最高,达到100%。多重耐药菌株YA-1全基因组测序证实,该菌株染色体大小为4 844 827 bp,携带3个完整的质粒,大小分别为144 776 bp(pTLYA-1)、113 584 bp(pTLYA-2)和77 544 bp(pTLYA-3)。分离菌株携带arr-3、aadA16、sul1等70个耐药基因,其中pTLYA-1中携arr-3、dfrA27、aadA16等8个可移动耐药基因,pTLYA-3中携带floR、tetA、sul2等5个可移动耐药基因,pTLYA-2中未发现耐药基因。【结论】由通辽地区部分鸭场分离的鸭源大肠杆菌存在多重耐药性,耐药水平较高,aphA1、strB和qnrS耐药基因普遍流行。

首次从湖南猪源大肠杆菌中检出mcr-1阳性IncI2(Delta)质粒

近年来,由于抗生素耐药基因的广泛传播及多重耐药细菌的出现,导致抗生素的使用面临严峻挑战,致使毒性较强的多黏菌素又重新引起业界的重视,为此对多黏菌素耐药情况的研究也变得十分重要。为了调查畜牧场中多黏菌素耐药基因mcr-1的流行情况,本研究选择我国湖南省某猪场240份肠肛拭子样品,培养分离鉴定出含有mcr-1耐药基因的大肠杆菌菌株,选取多黏菌素等10种抗菌药物对分离的菌株进行耐药性试验,对所有含mcr-1耐药基因的大肠杆菌阳性株进行全基因组测序(whole-genome sequencing,WGS),采用多位点序列分型(multi-locus sequence typing,MLST)技术进行大肠杆菌的遗传多样性分析。研究结果表明,在分离出的172株大肠杆菌中,来自不同个体的9株携带mcr-1基因的大肠杆菌均表现出多重耐药现象;MLST分析共鉴定出ST10、ST196、ST46和ST5229共4种分型和4种血清型(O83:H5,O16:H7,O16:H51,O9:H4)。生物信息学分析显示,所有阳性菌株均未发现与mcr-1基因传播有关的典型可移动遗传元件ISApl1,但均检出可能会导致mcr-1传播的IV型分泌系统基因。质粒接合转移试验与单倍型的MLST多态性结果表明,mcr-1基因主要以质粒介导的水平传播为主。本研究是国际上首次在猪源细菌中检出常见于人医临床细菌的携带有mcr-1抗性基因的IncI2(Delta)质粒。