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.

Burden of Multidrug-Resistant Escherichia coli in Pigs Slaughtered in Uganda and Its Implication on Veterinary Public Health

Antimicrobial resistance by bacteria and other microbes has become a global public and animal health threat. In this cross-sectional study, assessed the abattoir workers’ practices regarding pork handling and we investigated antimicrobial susceptibility patterns of Escherichia coli isolated from pigs brought for slaughter at Wambizzi, Uganda’s main pig abattoir. Rectal swabs were collected from a total of 176 live pigs prior to slaughter. Additionally, 24 swabs were taken from the abattoir floor environment. The collected swabs were cultured for the detection and isolation of E. coli followed by antibiotic susceptibility tests. Regarding pork handling practices, absence of hand washing facilities was observed and none of the workers cleaned/disinfected their equipment between slaughters while slaughters took place on the unhygienic floors of the inspection room. Overall, high prevalence (85.1%) of multi-drug resistant E. coli was detected in pigs received from all the regions of Uganda. Swine E. coli isolates exhibited high resistance against erythromycin (87.4%) and the least resistance against ciprofloxacin at 2.3%. At regional level, E. coli isolates from the central region of Uganda showed higher prevalence of multidrug resistant E. coli isolates as follows;amoxicillin (30.4%, p-value = 0.007), erythromycin (34.8%, p-value = 0.002), streptomycin (40.7%), ciprofloxacin (100%), oxytetracycline (31%) and sulphamethoxazole-trimethoprim (42.9%). Furthermore, multidrug-resistant E. coli was also confirmed in the immediate environment where pigs were gathered and slaughtered. From these environmental isolates, the highest resistance was confirmed against erythromycin (100%), whereas no isolates showed resistance against ciprofloxacin. The observed practices coupled with the presence of multidrug-resistant E. coli in the slaughterhouses presents a possible risk of pork contamination with multidrug-resistant E. coli presenting a potential risk of causing foodborne illnesses among pork consumers in Uganda. The current findings could justify active surveillance of antimicrobial resistance among food animals and provides basis for monitoring the quality of pork products to ensure food safety.

How Growth Ability of Multidrug-Resistant Escherichia coli Is Affected by Abiotic Stress Factors

The ability of multidrug-resistant Escherichia coli to adapt and grow in a wide range of different environmental conditions may be crucial to the global spread of antimicrobial resistance. The aim of this study was to evaluate the survival ability of 54 multidrug-resistant E. coli strains, isolated from three different biotopes (clinical setting, gull intestine, river water) when subjected to variations in pH (from 3 to 11) and salinity (from 0.5% to 6% of NaCl) and to nutrient deprivation. The growth of each isolate as well as of a reference strain was assessed during 168 h in every varying condition. Slight variations in the growth ability under some abiotic stress factors were recorded among the isolates from the different biotopes. Multidrug-resistant isolates from gull feces were found to be the more tolerant to environmental abiotic changes, while isolates from river water were the less tolerant. In addition, it was notorious that the carriage of antimicrobial resistance has a clear fitness cost in comparison with the susceptible (reference) strain, highlighting the necessity of reducing the selective pressure exerted by antibiotics. This study underlines the ecological hardness of multidrug-resistant E. coli isolates with a consequent ability to reach and colonize new host and environments.

Multidrug-Resistant of Escherichia coli and Salmonella spp. Strains in Chicken Feces Intended for Consumption in Open Spaces of Ouagadougou, Burkina Faso

Resistant bacteria can be transmitted to humans through feces or contaminated meat from local chickens. Bacterial strains were isolated from the intestinal contents of 400 local chicken samples from various sales sites. These strains were then characterized using bacteriological and biochemical methods to identify resistant strains. In a study conducted in Ouagadougou, we systematically collected chicken fecal samples from 20 locations across the city, followed by isolation and identification of Salmonella spp. using specific enrichment and culture methods, as well as Escherichia coli. Bacterial strains were characterized using antibiotic resistance profiles were determined through agar diffusion tests, revealing sensitivity or resistance to a range of antibiotics based on established scientific criteria. The results showed that out of the 400 samples collected, 81.25% and 63.5% were contaminated by Escherichia coli and Salmonella spp., respectively. Among these, 86.15% of identified Escherichia coli and 50.78% of Salmonella spp. displayed resistance to at least one tested antibiotic. Among 280 Escherichia coli isolates identified resistant to at least one antibiotic, 31.07% were resistant to cefotaxime (CTX), 20.35% to ceftazidime (CAZ), 21.07% to ceftriaxone (CTR), 75% to amoxicillin clavulanic acid (AMC), 23.57% aztreoname (ATM) and 27.14% were resistant to imipenem (IMP). In the case of the 129 Salmonella spp. isolates resistant to at least one tested antibiotic, 34.88% were resistant to CTX;41.08% to CAZ;35.65% to CTR, 92% to AMC, 39.53% to ATM and finally 47.28% were resistant to IMP. Our study revealed high prevalence of resistance in bacterial strains isolated from local chickens sold outdoors in Ouagadougou. These findings raise significant public health concerns, due to the possible transmission of these resistant strains to humans through the consumption of contaminated meat, thus complicating the treatment of bacterial infections.

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.

Discovery of natural berberine-derived nitroimidazoles as potentially multi-targeting agents against drug-resistant Escherichia coli

A series of natural berberine-derived nitroimidazoles as novel antibacterial agents were designed, synthesized and characterized by nuclear magnetic resonance(NMR), infrared spectra(IR), and high resolution mass spectra(HRMS) spectra. The antimicrobial evaluation showed that some target molecules exhibited moderate to good inhibitory activities against the tested bacteria and fungi including clinical drug-resistant strains isolated from infected patients. Especially, 2-fluorobenzyl derivative8 f not only gave strong activity against drug-resistant E. coli with the minimal inhibitory concentration(MIC) value of0.003 m M, 33-fold more active than norfloxacin, but also exhibited low toxicity toward RAW 264.7 cells and less propensity to trigger resistance. The aqueous solubility and Clog P values of target compounds were investigated to elucidate the structureactivity relationships. Molecular docking and quantum chemical studies for compound 8 f rationally explained its antibacterial effect. The further exploration of antibacterial mechanism revealed that the highly active compound 8 f could effectively permeabilize E. coli cell membrane and intercalate into DNA isolated from resistant E. coli to form 8 f-DNA complex that might block DNA replication to exert the powerful bioactivities. Compound 8 f could also selectively address resistant E. coli from a mixture of various strains.

Antibacterial mechanism of kojic acid and tea polyphenols against Escherichia coli O157:H7 through transcriptomic analysis

Escherichia coli O157:H7 is one of the major foodborne pathogenic bacterial that cause infectious diseases in humans.The previous found that a combination of kojic acid and tea polyphenols exhibited better activity against E.coli O157:H7 than using either alone.This study aimed to explore responses underlying the antibacterial mechanisms of kojic acid and tea polyphenols from the gene level.The functional enrichment analysis by comparing kojic acid and tea polyphenols individually or synergistically against E.coli O157:H7 found that acid resistance systems in kojic acid were activated,and the cell membrane and genomic DNA were destructed in the cells,resulting in“oxygen starvation”.The oxidative stress response triggered by tea polyphenols inhibited both sulfur uptake and the synthesis of ATP,which affected the bacteria's life metabolic process.Interestingly,we found that kojic acid combined with tea polyphenols hindered the uptake of iron that played an essential role in the synthesis of DNA,respiration,tricarboxylic acid cycle.The results suggested that the iron uptake pathways may represent a novel approach for kojic acid and tea polyphenols synergistically against E.coli O157:H7 and provided a theoretical basis for bacterial pathogen control in the food industry.

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.

Preclinical and clinical evidence of the association of colibactinproducing Escherichia coli with anxiety and depression in colon cancer

BACKGROUND The association between the intestinal microbiota and psychiatric disorders is becoming increasingly apparent.The gut microbiota contributes to colorectal carcinogenesis(CRC),as demonstrated with colibactin-producing Escherichia coli(CoPEC).AIM To evaluate the association between CoPEC prevalence and anxiety-and depressive-like behaviors with both preclinical and clinical approaches.METHODS Patients followed after a CRC surgery and for whom the prevalence of CoPEC has been investigated underwent a psychiatric interview.Results were compared according to the CoPEC colonization.In parallel C57BL6/J wild type mice and mice with a CRC susceptibility were chronically infected with a CoPEC strain.Their behavior was assessed using the Elevated Plus Maze test,the Forced Swimming Test and the Behavior recognition system PhenoTyper®.RESULTS In a limited cohort,all patients with CoPEC colonization presented with psychiatric disorders several years before cancer diagnosis,whereas only one patient(17%)without CoPEC did.This result was confirmed in C57BL6/J wildtype mice and in a CRC susceptibility mouse model(adenomatous polyposis colimultiple intestinal neoplasia/+).Mice exhibited a significant increase in anxiety-and depressive-like behaviors after chronic infection with a CoPEC strain.CONCLUSION This finding provides the first evidence that CoPEC infection can induce microbiota-gut-brain axis disturbances in addition to its procarcinogenic properties.

The virulence regulator AbsR in avian pathogenic Escherichia coli has pleiotropic effects on bacterial physiology

Avian pathogenic Escherichia coli(APEC)belonging to extraintestinal pathogenic E.coli(ExPEC)can cause severe infections in extraintestinal tissues in birds and humans,such as the lungs and blood.MprA(microcin production regulation,locus A,herein renamed AbsR,a blood survival regulator),a member of the MarR(multiple antibiotic resistance regulator)transcriptional regulator family,governs the expression of capsule biosynthetic genes in human ExPEC and represents a promising druggable target for antimicrobials.However,a deep understanding of the AbsR regulatory mechanism as well as its regulon is lacking.In this study,we present a systems-level analysis of the APEC AbsR regulon using ChIP-Seq(chromatin immunoprecipitation sequencing)and RNA-Seq(RNA sequencing)methods.We found that AbsR directly regulates 99 genes and indirectly regulates 667 genes.Furthermore,we showed that:1)AbsR contributes to antiphagocytotic effects by macrophages and virulence in a mouse model for systemic infection by directly activating the capsular gene cluster;2)AbsR positively impacts biofilm formation via direct regulation of the T2SS(type II secretion system)but plays a marginal role in virulence;and 3)AbsR directly upregulates the acid tolerance signaling system EvgAS to withstand acid stress but is dispensable in ExPEC virulence.Finally,our data indicate that the role of AbsR in virulence gene regulation is relatively conserved in ExPEC strains.Altogether,this study provides a comprehensive analysis of the AbsR regulon and regulatory mechanism,and our data suggest that AbsR likely influences virulence primarily through the control of capsule production.Interestingly,we found that AbsR severely represses the expression of the type I-F CRISPR(clustered regularly interspaced short palindromic repeats)-Cas(CRISPR associated)systems,which could have implications in CRISPR biology and application.

Prevalence of Drug Resistant Uropathogenic Escherichia coli from Immunocompromised Diabetic Patients Attending Selected Health Facilities in Benue State

Escherichia coli is the commonest bacterial uropathogen of UTIs, the commonest infections in immunocompromised diabetic patients. Better understanding of their main resistance mechanisms to commonly used antibacterial agents will help to reduce the burden of this infection. The prevalence of drug resistant uropathogenic Escherichia coli isolates from immunocompromised diabetic patients attending selected health facilities in Benue State was investigated. Two hundred and ninety-six midstream urine samples were collected for both study and control diabetic patients. Bacterial isolation was done using semi-quantitative method. Drug resistant Escherichia coli were identified as multidrug resistant (MDR), extensive drug resistant (XDR) and pan-drug resistant organisms (PDR). Statistical significance was considered at p E. coli isolates from the study and control subjects with overall prevalence of 20.9% and 8.4% respectively. The isolates were highly resistant to penicillin (ampicillin), monobactam (aztreonam), older quinolone (nalidixic acid) whereas the majority of them showed high susceptibility to aminoglycoside (streptomycin), cephalosporin (cefotaxime) and carbapenem (imipenem). None showed complete susceptibility to all the tested antibiotics. Twenty-five E. coli were identified in this MDR, eight, XDR while 5 were PDR. High numbers of drug resistant E. coli isolates were identified in the study group of which 25 were MDR, 8 XDR while 5 were PDR isolates. High prevalence of UTI and drug resistant isolates occur in diabetic patients with hyperglycemic condition.

Impact of Genetic Diversity of Uropathogenic Escherichia coli and Klebsiella pneumoniae Strains on the Dissemination of Extended Spectrum Beta-Lactam Resistance Genes in Côte d’Ivoire

The increase and spread of bacterial resistance to extended-spectrum beta-lactam antibiotics are reported in many infections and are a real public health problem worldwide. Drug pressure is a factor that favors the emergence of a population of better adapted bacteria. However, there is no literature highlighting the genetic diversity and evolutionary structure of E. coli and K. pneumoniae in an environment with high selection pressure in Côte d’Ivoire. The objective of this study was to evaluate the genetic diversity of E. coli and K. pneumoniae strains circulating at the HKB Hospital in Abobo and at the Daloa Regional Hospital and its impact on the dissemination of extended spectrum beta-lactam resistance genes. A total of 39 strains isolated from the urinary tract of infected patients, including 30 strains of E. coli and 9 strains of K. pneumoniae were studied. A total of 39 strains isolated from the urinary tract of infected patients, including 30 strains of E. coli and 9 strains of K. pneumoniae were studied. From genomic DNA extracts, ESBL resistance genes were amplified by PCR and sequenced, in addition to genetic typing by ERIC-PCR. The data obtained were submitted to genetic and bioinformatics analyses. The results have shown a genetic diversity important in E. coli and K. pneumoniae with diversity indexs (SID) ranging from 0.5 to 0.77. The genetic structure of the bacterial species studied has shown a clonal distribution of strains with clones expressing TEM-9 and CTX-M-15 variants. Also, this clonal structure was correlated with the spread of resistance genes in E. coli and K. pneumoniae. The spread of resistant clones is a factor that might limit the fight against antibiotic resistance.

Phytochemicals of Aloe barbadensis miller as Potential Inhibitors of Uropathogenic Escherichia coli for Urinary Tract Infection Therapy: An in Silico Approach

Urinary tract infections (UTIs) are common infections caused by normal skin or rectum bacteria that get into the urethra and infect the urinary tract. Although the infection can affect various parts of the tract, bladder infections are the most prevalent kind. Uropathogenic Escherichia Coli (UPEC) is the most common pathogen associated with UTI development. Therefore, inhibiting the UPEC protein target (PDB ID: 8BVD) appears to be a promising therapeutic strategy. Therefore, in this study, molecular docking and dynamics were conducted to examine the antibacterial activity of Aloe barbadensis miller against UPEC bacteria. The Aloe barbadensis miller natural compounds licochalcone A, palmidin B and palmidin C were downloaded from PubChem with amoxicillin, which was used as a control drug and studied against the target molecule. The potential parameters examined were the docking scores, absorption, distribution, metabolism, excretion, toxicity (ADMET), bioavailability, root mean square deviation (RMSD), root mean square fluctuation (RMSF), hydrogen bonding, radius of gyration, and potential energy of the system. Docking scores showed that all ligands demonstrated an admirable candidature as an inhibitor to 8BVD molecule, and the score hierarchy is licochalcone A (-6.4 kcal/mol), palmidin C (-6.1 kcal/mol), palmidin B (-6.0 kcal/mol), and amoxicillin (-5.9 kcal/mol). All ligands appeared to have good drug-like properties and oral bioavailability. Molecular dynamic studies showed that all ligands exhibited an excellent nominee as inhibitors in their vicinity at 20 ns. However, there is a relatively high fluctuation of palmidin B compared with other compounds, which seems to be more stable. This work suggests that the selected phytoconstituents could be used as inhibitors of the 8BVD protein in the fight against UTIs. However, further investigation on the clinical and experimental validation of UTI treatment’s specific mechanisms and effects is still welcomed.

Presence of Multidrug-Resistant <i>E. coli</i>, <i>Enterococcus</i>spp. and <i>Salmonella</i>spp. in Lakes and Fountains of Porto, Portugal

Urban lakes and fountains provide recreational activities that could facilitate the contact between humans, animals and biological agents. The objective of this work was to assess the water quality and safety of 17 lakes and 13 fountains in the city of Porto (Portugal), by detecting the presence of Escherichia coli, enterococci and Salmonella spp., and analyzing their antimicrobial resistance. (For more information,please refer to the pdf.)

Sterilization Effects of Bacterial Inhibitor on Escherichia coli in Cattle Manure Compost

[Objective] The aim of this study was to explore the sterilization effects on Escherichia coli by adding bacterial inhibitor(CaCN2)during the process of cattle manure composting so as to provide a theoretical basis for cattle manure harmless treatment.[Method] Both experimental groups supplemented with 2.0% bacterial inhibitor and control groups without bacterial inhibitor were cultured under different temperatures(20,30,37,50,60 ℃)to determine the optimal composing temperature.Under 30 ℃,different bacterial inhibitor doses(0,2.0%,2.5%,3.0%)were added into the compost to obtain the optimal bacterial inhibitor addition dose.[Result] 30,50 and 60 ℃ were ideal temperatures for sterilization of E.coli.Under 30 ℃,E.coli couldn't be detected in 2.5% dose group and 3.0% dose group after culture for 48 h,demonstrating no less than 2.5% bacterial inhibitor should be added.[Conclusion] It has an important significance to enhance the sterilization effects on E.coli by adding CaCN2 into cattle manure compost especially in winter.

口服益生菌Escherichia coli Nissle 1917调控断奶仔猪空肠黏膜屏障功能的机理研究

旨在探讨口服益生菌Escherichia coli(E.coli)Nissle 1917调控病原菌E.coli Abbottstown攻毒或未攻毒21d断奶仔猪肠道屏障功能的机理。将20头21d断奶仔猪((6.35±0.38)kg)随机分为4组:(1)饲喂基础日粮的对照组(CON);(2)正试期口服E.coli Nissle 1917组(EcN);(3)预试期E.coli Abbottstown攻毒组(EcA);(4)预试期E.coli Abbottstown攻毒和正试期口服E.coli Nissle 1917组(EcN+EcA)。每个处理5个重复,每个重复1头仔猪。试验分为3d预试期和21d正试期。预试期EcA和EcN+EcA组的断奶仔猪接种5×10~9E.coli Abbottstown。正试期EcN和EcN+EcA组仔猪每天每头口服1×10~10E.coli Nissle 1917。试验结果表明,未攻毒的断奶仔猪中,与CON相比,口服1×10~10E.coli Nissle 1917能显著改善生长性能(P<0.05),降低腹泻率,提高oCcludin蛋白水平(P<0.05),显著降低空肠黏膜calprotectin蛋白水平(P<0.05),显著下调空肠黏膜蛋白激酶R(Protein kinase R,PKR)、toll样受体-2(Toll-like receptor-2,TLR-2)和真核起始因子-5A(eukaryotic initiation factor-5A,eIF-5A)mRNA的相对表达丰度(P<0.05),上调空肠黏膜内皮生长因子(Endothelial growth factor,EGF)、肝细胞生长因子(Hepatocyte growth factor,HGF)、胰岛素样生长因子(Insulin-like growth factor,IGF-I)、三叶肽因子-3(trefoil peptide factor-3,TFF-3)mRNA的相对表达丰度(P<0.05);在E.coli Abbottstown攻毒的断奶仔猪中,与EcA组相比,口服1×10~10E.coli Nissle 1917能显著改善生长性能(P<0.05),降低腹泻率,降低血清二胺氧化酶(DAO)活性(P<0.05)和一氧化氮(NO)含量(P<0.05),下调空肠黏膜PKR、eIF-5A和TLR-2 mRNA相对表达丰度(P<0.05),显著降低空肠黏膜calprotectin蛋白水平和淋巴细胞数量(P<0.05),显著提高空肠黏膜oCcludin蛋白水平(P<0.05),上调空肠黏膜IGF-I、HGF、TFF-3和EGF mRNA相对表达丰度(P<0.05)。本试验结果提示,口服1×10~10E.coli Nissle 1917能降低病原大肠杆菌攻毒或未攻毒21日龄断奶仔猪空肠黏膜屏障功能的损伤。

硝酸镧引起Escherichia coli B代谢过程热爆发及其机理研究

采用微量热法研究了硝酸镧对Escherichia coli B生长代谢过程的影响,发现高浓度硝酸镧引起E.coli B热谱图出现异常变化:生长速率常数k值增大、产热峰显著升高和总发热量异常增加.当硝酸镧浓度为300和500mg/L时,培养物在培养过程的总发热量分别是正常条件下的3.89和2.54倍.用生物学方法对细胞存活率和生物量进行测定结果表明,细胞在高浓度硝酸镧条件下增殖受到抑制、细胞生物量减少.表明高浓度的硝酸镧存在时,E.coli B细胞生长受到抑制反而释放出比正常生长细胞多得多的热量,将抑制状态细胞释放大量热量的现象称为热爆发.分析热爆发的原因,认为是La3+离子破坏细胞壁外膜而增加其透性,导致细胞膜与外膜间的质子电化学势因质子外泄而降低或者不能形成,氧化磷酸化过程中的能量不能有效地转化为ATP,而以热能的方式释放出来.细胞由于缺乏生物通用能量ATP,因而其生长受到抑制.

Morphological Changes of Hemocytes of Musca domestica Larva in vitro Infected by Escherichia coli

[Objective] The research aimed to observe the effects of Escherichia coli infection on the morphology of hemocytes of the 3rd stage larva of Musca domestica in vitro and understand the hemocytes types that take part in the cell immunity of Musca domestica larval.[Method] The hemcytes of the 3rd stage larva of Musca domestica were cultured in vitro and the hemocyte morphology was observed about 2,4,6,8 h after culture in vitro.After Escherichia coli were injected into the hemocytes of the 3rd stage larva of Musca domestica in vitro,the morphology changes of hemocytes were observed at different time after infection.[Result] The hemocytes of of the 3rd stage larva of Musca domestica was divided into five types about 2 h after hemoculture.The hemocytes partly adherence was seen about 6 h after hemoculture.The vacuolation and morpholysis was found in plasmatocytes after being infected by E.coli and a great quantity bacterium were gathered around granulocyte,but the morphology changes of hemocytes were not found in the prohemocyte,shprulocyte and oenocytoid.[Conclusion] The plasmatocyte and granulocyte were primary participants of the cell immunity of Musca domestica larval,but the prohemocyte,sphrulocyte and oenocytoid do not participate in the cell immune reactions.

PCR Detection of Virulence Genes Colv,Stxs and HlyE of Escherichia coli

[Objective] This study aimed to explore the presence of three causative genes Colv,Stxs and HlyE of the pathogenic E.coli from chickens,pigs and food.[Method] By using 44 E.coli strains from chickens,24 from pigs and 26 from food as the experimental materials,virulence genes Colv,Stxs（stx2,stx2e） and HlyE were detected with polymerase chain reaction（PCR） method.[Result] Among all the E.coli strains,the detection rate of Colv was 25% from chickens,4.2% from pigs,and 0 from food;the detection rate of Stx2（Stx2e） from all E.coli strains was 0;the detection rate of HlyE was 2.27% from chickens,0 from pigs,and 11.5% from food.[Conclusion] Virulence gene Colv shows relatively high carrying rate in E.coli from chickens and pigs;HlyE also shows a certain degree of presence in E.coli from chickens and food.

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.