Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to ...Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to carbadox.The objective of this study was to investigate the impacts of oligosaccharide-based polymer or antibiotic on the host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.Results Multivariate analysis highlighted the differences in the metabolic profiles of serum and colon digesta which were predominantly found between pigs supplemented with oligosaccharide-based polymer and antibiotic.The relative abundance of metabolic markers of immune responses and nutrient metabolisms,such as amino acids and carbohydrates,were significantly differentiated between the oligosaccharide-based polymer and antibiotic groups(q<0.2 and fold change>2.0).In addition,pigs in antibiotic had a reduced(P<0.05)relative abundance of Lachnospiraceae and Lactobacillaceae,whereas had greater(P<0.05)Clostridiaceae and Streptococcaceae in the colon digesta on d 11 post-inoculation(PI)compared with d 5 PI.Conclusions The impact of oligosaccharide-based polymer on the metabolic and microbial profiles of pigs is not fully understood,and further exploration is needed.However,current research suggest that various mechanisms are involved in the enhanced disease resistance and performance in ETEC-challenged pigs by supplementing this polymer.展开更多
BACKGROUND:The virulent factors of Escherichia coli(E.coli) play an important role in the process of pathopoiesis.The study aimed to compare drug-resistant genes and virulence genes between extended spectrum β-lactam...BACKGROUND:The virulent factors of Escherichia coli(E.coli) play an important role in the process of pathopoiesis.The study aimed to compare drug-resistant genes and virulence genes between extended spectrum β-lactamases(ESBLs)-producing E.coli and non-ESBLs-producing E.coli to provide a reference for physicians in management of hospital infection.METHODS: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,qacEA1-sull and 8 virulence genes(CNF2,hlyA,eaeA,VT1,est,bfpA,elt,and CNF1) were tested by polymerase chain reaction(PCR).RESULTS:Among the 96 E.coli isolates,the ESBLs-producing E.coli comprised 46(47.9%)strains and the non-ESBLs-producing E.coli consisted of 50(52.1%) strains.The detection rates of multiple drug-resistant strain,qacEA1-sull,CNF2,hlyA,eaeA,VT1,est,bfpA,elt,and CNF1 in 46ESBLs-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.coli strains,the positive rates of multiple drug-resistant strain,qacEA1-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.coli strains and the non-ESBLs-producing E.coli strains was statistically significant(P<0.05).CONCLUSION:The positive rate of multiple drug-resistant strains is higher in the ESBLsproducing 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.展开更多
Background: Recently micro-organisms that synthesize extended-spectrum β-lactamase (ESBLs) were increased. The peculiarities of ESBL synthesis of Escherichia coli and Klebsiella pneumoniae strains that cause nosocomi...Background: Recently micro-organisms that synthesize extended-spectrum β-lactamase (ESBLs) were increased. The peculiarities of ESBL synthesis of Escherichia coli and Klebsiella pneumoniae strains that cause nosocomial urinary tract infections, surgical site infections and pneumonia in surgical clinic were studied. ESBL synthesis were observed 38.9% of E. coli strains obtained from urine, 92.3% of strains obtained from surgical site infections, and 50% of strains obtained from sputum. ESBL synthesis were observed 37.5% of K. pneumoniae strains obtained from urine, 85.7% of strains obtained from surgical site infections, and 60% of strains obtained from sputum. Different levels of ESBL synthesize of E. coli and K. pneumoniae strains isolated from different pattern is discussed. Conclusion. ESBL synthesis is common in E. coli and K. pneumoniae strains, which cause nosocomial infections. The frequency of occurrence of ESBL s synthesis among of these strains depends on clinical forms of nosocomial infections.展开更多
Fecal coliform bacteria such as Escherichia coli (E. coli) are one of the main sources of groundwater pollution. An assessment of the transport and Persistence of E. coli in poultry litter amended Decatur silty Clay s...Fecal coliform bacteria such as Escherichia coli (E. coli) are one of the main sources of groundwater pollution. An assessment of the transport and Persistence of E. coli in poultry litter amended Decatur silty Clay soil and Hartsells Sandy soil was conducted using soil columns and simulated groundwater leaching. Enumeration of initial E. coli was determined to range from 2.851 × 10<sup>3</sup> to 3.044 × 10<sup>3</sup> CFU per gram of soil. These results have been used in a batch study to determine the persistence rate of E. coli in Decatur silty Clay soil and Hartsells Sandy soil. Results prove that E. coli survival growth rate increases for clay soil later than and at a higher rate than sandy soil. The column study has determined that E. coli was transported at a rate of 3.7 × 10<sup>6</sup><sup> </sup>CFU for Decatur silty loam and 6.3 × 10<sup>6</sup><sup> </sup>CFU for Hartsells sandy per gram of soil. Further, linear regression analysis predictions show higher porosity and soil moisture content affect transport, and Hartsells sandy soil has higher transport of E. coli due to its higher porosity and lower volumetric water content.展开更多
A preliminary study was made on the antibacterial mechanism of copper-bearing antibacterial stainless steels against E.coli through experiments of microbiology such as EDTA (ethylenediaminetetraacetic acid) complexi...A preliminary study was made on the antibacterial mechanism of copper-bearing antibacterial stainless steels against E.coli through experiments of microbiology such as EDTA (ethylenediaminetetraacetic acid) complexing, DNA smearing and AFM (atomic force microscope) observation. It was measured that the antibacterial stainless steels showed excellent antibacterial functions with antibacterial rate to E.coli over 99.99%. The antibacterial rate was weak if the bacteria solution was complexed by EDTA, indicating that the copper ions play a dominant role in the antibacterial effect of the antibacterial stainless steels. The electrophoresis experiment did not show the phenomenon of DNA smearing for E.coli after contacting antibacterial stainless steels, which meant that DNA of E.coli was not obviously damaged. It was observed by AFM that the morphology of E.coli changed a lot after contacting antibacterial stainless steels, such as cell walls being seriously changed and lots of contents in the cells being leaked.展开更多
Background:The independent and interactive effects of dietary fiber(DF)and threonine(Thr)were investigated in growing pigs challenged with either systemic E.coli lipopolysaccharide(LPS)or enteric Salmonella Typhimuriu...Background:The independent and interactive effects of dietary fiber(DF)and threonine(Thr)were investigated in growing pigs challenged with either systemic E.coli lipopolysaccharide(LPS)or enteric Salmonella Typhimurium(ST)to characterise their effect on intestinal barrier function.Results:In experiment 1,intestinal barrier function was assessed via oral lactulose and mannitol(L:M)gavage and fecal mucin analysis in pigs challenged with E.coli LPS and fed low fiber(LF)or high fiber(HF)diets with graded dietary Thr.Urinary lactulose recovery and L:M ratio increased(P<0.05)during the LPS inoculation period in LF fed pigs but not in HF fed pigs.Fecal mucin output was increased(P<0.05)in pigs fed HF compared to LF fed pigs.In experiment 2,RT-qPCR,ileal morphology,digesta volatile fatty acid(VFA)content,and fecal mucin output were measured in Salmonella Typhimurium challenged pigs,fed LF or HF diets with standard or supplemented dietary Thr.Salmonella inoculation increased(P<0.05)fecal mucin output compared to the unchallenged period.Supplemental Thr increased fecal mucin output in the HF-fed pigs(Fib×Thr;P<0.05).Feeding HF increased(P<0.05)VFA concentration in cecum and colon.No effect of either Thr or fiber on expression of gene markers was observed except a tendency(P=0.06)for increased MUC2 expression with the HF diet.Feeding HF increased goblet cell numbers(P<0.05).Conclusion:Dietary fiber appears to improve barrier function through increased mucin production capacity(i.e.,goblet cell numbers,MUC2 gene expression)and secretion(i.e.,fecal mucin output).The lack of effect of dietary Thr in Salmonella-challenged pigs provides further evidence that mucin secretion in the gut is conserved and,therefore,Thr may be limiting for growth under conditions of increased mucin production.展开更多
Objective To report a protocol using biotin-labelled PrP protein in cell free conversion assay instead of isotope. Methods A hamster PrP protein (HaPrP) was expressed in E. coli and purified with HIS-tag affinity ch...Objective To report a protocol using biotin-labelled PrP protein in cell free conversion assay instead of isotope. Methods A hamster PrP protein (HaPrP) was expressed in E. coli and purified with HIS-tag affinity chromatograph. After being labelled with biotin, HaPrP was mixed with PrP^sen preparation from scrapie strain 263K. Results Protease-resistant bands were detected after four-day incubation. Conclusion The new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.展开更多
AIM: To study the effect of integration of tandem aroG-pheA genes into the tyrA locus of Corynebacterium glutamicum (C. glutamicum) on the production of L-phenylalanine.METHODS: By nitrosoguanidine mutagenesis, five p...AIM: To study the effect of integration of tandem aroG-pheA genes into the tyrA locus of Corynebacterium glutamicum (C. glutamicum) on the production of L-phenylalanine.METHODS: By nitrosoguanidine mutagenesis, five pfluorophenylalanine (FP)-resistant mutants of Cglutamicum FP were selected. The tyrA gene encoding prephenate dehydrogenase (PDH) of C.glutamicum was amplified by polymerase chain reaction (PCR) and cloned on the plasmid pPR. Kanamycin resistance gene (Km) and the PBF-aroGpheA-T (GA) fragment of pGA were inserted into tyrA gene to form targeting vectors pl-K and pTGAK, respectively. Then,they were transformed into C.glutamicum FP respectively by electroporation. Cultures were screened by a medium containing kanamycin and detected by PCR and phenotype analysis. The transformed strains were used for L-phenylalanine fermentation and enzyme assays.RESULTS: Engineering strains of C.glutamicum (Tyr-)were obtained. Compared with the original strain, the transformed strain C. glutamicum GAK was observed to have the highest elevation of L-phenylalanine production by a 1.71-fold,and 2.9-, 3.36-, and 3.0-fold in enzyme activities of chorismate mutase, prephenate dehydratase and 3-deoxy-Darabinoheptulosonate-7-phosphate synthase, respectively.CONCLUSION: Integration of tandem aroG-pheA genes into tyrA locus of C. glutamicum chromosome can disrupt tyrA gene and increase the yield of L-phenylalanine production.展开更多
Background: This experiment was conducted to test the hypothesis that vitamin E(Vit E) and acetylsalicylic acid(ASA), a cyclooxygenase-2(COX-2) inhibitor, will additively reduce the production of the immunosupp...Background: This experiment was conducted to test the hypothesis that vitamin E(Vit E) and acetylsalicylic acid(ASA), a cyclooxygenase-2(COX-2) inhibitor, will additively reduce the production of the immunosuppressive molecule prostaglandin E_2(PGE_2) and hence reduce inflammatory responses in weaner pigs experimentally infected with an enterotoxigenic strain of E. coli.Methods: The experiment was conducted in a research facility with 192 individually-housed male weaner pigs(Landrace × Large White) weighing 6.6 ± 0.04 kg(mean ± SEM). The pigs were experimentally infected with an enterotoxigenic strain of E. coli and were allocated to a 2 × 3 factorial design with the respective factors being without and with 125 ppm ASA and three levels of Vit E supplementation(50, 100 or 200 IU/kg diet, dl-α-tocopheryl acetate).Results: Acetylsalicylic acid supplementation improved average daily gain(P 〈 0.05) and tended to improve feed:gain ratio(P 〈 0.10) during the first 14 d after weaning. Acetylsalicylic acid supplementation also improved(P 〈 0.001) amino acid utilization efficiency(as assessed by plasma urea level) and tended to decrease(P 〈 0.10) PGE2 production in the liver without affecting smal intestinal histology and tight junction protein mR NA expression in the jejunal epithelium. Vitamin E supplementation greater than 100 IU/kg diet sustained both the plasma Vit E concentration(P 〈 0.001) and plasma haptoglobin content(P 〈 0.001) after weaning. However, there was no additive effects of the combined supplementation of ASA and Vit E on performance, intestinal barrier function and inflammatory responses of weaned pigs.Conclusions: Although ASA and vitamin E improved amino acid utilization efficiency and reduced acute inflammatory responses, ASA and vitamin E did not additively reduce production of PGE2 and inflammatory responses in weaner pigs experimental y infected with an enterotoxigenic strain of E. coli.展开更多
Objective To investigate whether the effect of E.coli on U937 cell lines apoptosis is mediated via p38 mitogen-activated protein kinase (MAPK) activation. Methods The U937 cell lines were treated with E.coli at differ...Objective To investigate whether the effect of E.coli on U937 cell lines apoptosis is mediated via p38 mitogen-activated protein kinase (MAPK) activation. Methods The U937 cell lines were treated with E.coli at different time or together with SB203580, an inhibitor for p38. Cell apoptosis was analyzed by flow cytometry. p38 activities were detected by Western blotting. Results E.coli induced apoptosis in cultured U937 cell lines in a time-dependent manner. The phosphorylation of p38 was induced after 10 minutes infection, reached the peak after 20 minutes, and started to decline after 30 minutes. In contrast, the level of total p38 protein was not changed in whole experimental period. Inhibition of p38 with SB203580 significantly inhibited E.coli induced apoptosis in U937 cells. Conclusion The activation of the p38 MAPK in U937 cell lines by E.coli is a major pathway to mediate the apoptosis.展开更多
Ciprofloxacin is a second-generation of fluoroquinolone,broad-spectrum antibiotic with bactericidal activity against Gram-positive and Gram-negative organisms.It is one of the most widely used antibiotics,because of i...Ciprofloxacin is a second-generation of fluoroquinolone,broad-spectrum antibiotic with bactericidal activity against Gram-positive and Gram-negative organisms.It is one of the most widely used antibiotics,because of its efficacy,safety,and relatively low cost.Ascorbic acid(vitamin C)is water-soluble monosaccharide antioxidant;it is essentially required by the body for its various biochemical and physiological processes.S.aureus is Gram-positive cocci;widely distributed in the environment,it is a member of the normal flora of the body.S.aureus is not always pathogenic;it is a common cause of skin infections including abscesses,respiratory infections such as sinusitis,and food poisoning.E.coli is Gram-negative bacteria,found in the environment,foods,and intestines.Most E.coli strains are harmless;it is part of the normal microbiota of the gut.However,some serotypes of E.coli cause serious food poisoning in their hosts;it can cause diarrhea,while others cause urinary tract infections,respiratory illness and pneumonia,and other illnesses.Method:Cup cut diffusion method was applied.Experiment I:is carried out to choose the concentration of vitamin C to be used in experiment II.The negative control is normal saline,added in cup in each plate,vitamin C(100 mg/mL,200 mg/mL,400 mg/mL)was added,the volume in each cup was 100μL.Experiment II:Eight groups of treatments were applied.The first is the negative control(1%normal saline),the second group is the positive control of vitamin C(200 mg/mL).The third,fourth and fifth groups are ciprofloxacin with different concentrations(10 mg/mL,20 mg/mL,40 mg/mL);the sixth,seventh and eighth are the combination of vitamin C with each concentration of ciprofloxacin(10 mg/mL,20 mg/mL,40 mg/mL).Each group includes six petri dishes.Bacterial plates were incubated at 37 oC for 24 h and 48 h.Zone of inhibition is measured in mm.Results and conclusion:Ciprofloxacin produces dose dependent increase in zone of inhibition of S.aureus and E.coli growth,after 24 and 48 hours incubation.While vitamin C in the concentration used produced inhibitory effect on the growth of S.aureus and E.coli,after 24 hours incubation,vitamin C effect was not changed after 48 hours incubation.After 24 hours incubation,vitamin C potentiated the effect of ciprofloxacin at low concentration(10 mg/mL);while vitamin C antagonized the effect of ciprofloxacin at higher concentrations(20 and 40 mg/mL)on S.aureus growth.In the same time,ciprofloxacin antagonized the inhibitory effect of vitamin C on S.aureus growth.After 48 hours incubation,S.aureus produced resistance against ciprofloxacin alone,and that combined with vitamin C compared to zone of inhibition after 24 hours.Ciprofloxacin produced dose dependent inhibition of E.coli growth after incubation for 24 and 48 hours.Vitamin C potentiated the inhibitory effect induced by ciprofloxacin(additive effect).The inhibitory effect of ciprofloxacin,vitamin C and the combination was not changed after 48 hours compared to 24 hours.展开更多
Objective To study MIC value of 7 boron derivatives namely [Boric acid (H3BO3), Anhydrous Borax (Na2B407), Sodium Borate (NaBO2), Diammonium Tetraborate (NH4)2B4O7, Sodium Perborate (NaBO3), Boron Trioxide ...Objective To study MIC value of 7 boron derivatives namely [Boric acid (H3BO3), Anhydrous Borax (Na2B407), Sodium Borate (NaBO2), Diammonium Tetraborate (NH4)2B4O7, Sodium Perborate (NaBO3), Boron Trioxide (B203), Potassium Tetraborate (K2B407)] on E. coil and P. aeruginosa and their effects on survival of bacteria in lake water and resistance against kanamycin antibiotic. Methods MIC values of Boron derivatives and antibiotic were studied by broth microdilution method. The effect of boron derivatives on survival of bacteria in lake water were also determined with plate count. Results Sodium perborate was determined as the substances. Effectiveness increased as temperature most effective substance among the studied increased. E. coil was more affected from P. aeruginosa in 8 mg/mL sodium perborate concentration in lake water. Moreover, it was determined that MIC value of kanamycin antibiotic decreased 200 times by especially treating P. aeruginosa with sodium perborate in lake water. However, it can be stated that this change in resistance did not arise from microorganisms. Conclusion Sodium perborate solution can be used supportedly in kanamycin antibiotic applications for P. aeruginosa. Future studies are necessary to explore the relation between sodium perborate and kanamycin which is effective on P. aeruginosa in lake water.展开更多
The Ca. LSU intron flanking a 129 bp cxon upstream and a fOO bp exondownstream was inserted into the lacZ gene on pRS426 to transform E. coli. Northern blot analysisand RT-PCR showed that splicing of Ca. LSU in E. col...The Ca. LSU intron flanking a 129 bp cxon upstream and a fOO bp exondownstream was inserted into the lacZ gene on pRS426 to transform E. coli. Northern blot analysisand RT-PCR showed that splicing of Ca. LSU in E. coli is efficient upon inducible expression of theprecursor RNA, In contrast, co-lranscriptional self-splicing of the intron in vitro is much lessactive. Therefore, this E. coli splicing system can be used as a better model to investigate theeffect of the ribozyme inhibitors on Ca. LSUsplicing in living cell. We examined (he effects ofneomycin sulfatc and pentamidine on Ca. LSU splicing in E. coli, and found that these drugsdoes-dependently inhibit the intron splicing. However, heomycin is more potent than pentamidine inthis action.展开更多
To obtain the recombinant core domain of porcine zone pellucida 3β (cZP3β) for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained ...To obtain the recombinant core domain of porcine zone pellucida 3β (cZP3β) for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained by PCR and then was cloned into pET-3c vector. After being identified, recon was transformed into E.coli BL21 (DE3) pLysS and then induced by IPTG. Results The recombinant cZP3β was expressed in E. coli up to 15% of total cellular proteins, and was made sure by Western blot analysis. Conclusion The research on expression of core domain of pZP3β could benefit to further investigation of its immunogenicity and the development of antigen preparation.展开更多
The photocatalytic degradation of E.coli membrane cell by ZnO nanowires was studied using field-emission scanning electron microscope(FE-SEM),fluorescence microscopy,and Attenuated total reflection fourier transform...The photocatalytic degradation of E.coli membrane cell by ZnO nanowires was studied using field-emission scanning electron microscope(FE-SEM),fluorescence microscopy,and Attenuated total reflection fourier transform infrared(ATR-FTIR).The outer membrane of E.coli was removed completely in the presence of ZnO nanowires under UV irradiation,and the cells became twisted shapes without a mechanically strong network.After ZnO nanowires photocatalysis,the permeability of the treated cells increased to some degree that could be confirmed by quantum dots labeling technique.Structural changes in the cell wall membrane were revealed by the decay of the characteristic groups bands in ATR-FTIR spectra.展开更多
Objective:Diarrhea is a leading cause of morbidity and mortality among children in developing countries.The bacterial pathogen most commonly associated with childhood diarrhea is Escherichia coli.A one-year prospectiv...Objective:Diarrhea is a leading cause of morbidity and mortality among children in developing countries.The bacterial pathogen most commonly associated with childhood diarrhea is Escherichia coli.A one-year prospective study was carried out in Sanandaj to determine the prevalence and roles of the different E.coli pathotypes in children less than five years of age with acute diarrhea.Methods:Rectal swab were collected prospectively from children with acute diarrhea and transported to the Department of Microbiology,School of Medicine, KUMS,Sanandaj during 2008.The study was approved by the institutional ethics committee.Results:During this study period,rectal swabs were investigated from a total of 466 children 1 to 144 months of age(mean, 29.97 months±S.D) with diarrhea.Among the children,191(41%,191/466) were girls,and 275 (59%,275/466) were boys.The age-specific incidence rates of acute diarrhea among children 13-24 and 1 - 12 months of age were 37.37%(37/99) and 26.26%(26/99),respectively,during the study period.A total of 99 strains of E.coli were detected.EPEC 59(59.59%) and EIEC 22(22.22%),were the most commonly found Escherichia coli strains detected in stools from children.Disk diffusion testing showed E.coli strains resistance to tetracycline(89.89%),chloramphenicol(88.88%),Ampicillin(79.79%),Amoxicillin (75.75%) and Ceficime(75.75%).Among risk factors like age,sex,haemoglubin,fathers and mothers education,food and weight of children only mother's education was significant(P =0.018).Conclusion: In most of the clinical laboratories in Iran,E.coli does not considered as an etiologic agent responsible for diarrhea. Results in this study revealed that E.coli should be considered as an etiologic agent causing acute diarrhea among children.We therefore,recommend the routine isolation and identification of E.coli strains in all the clinical laboratories in Sanandaj.Guidelines for appropriate use of antibiotics in Sanandaj need updating.展开更多
[Objectives] The study aimed to identify the pathogenic E. coli strain that caused diarrhea in foxes and to analyze its drug sensitivity.[Methods] A pathogenic E. coli strain was isolated from dead foxes with diarrhea...[Objectives] The study aimed to identify the pathogenic E. coli strain that caused diarrhea in foxes and to analyze its drug sensitivity.[Methods] A pathogenic E. coli strain was isolated from dead foxes with diarrhea. By conventional bacterial isolation and culture, morphological observation, pathogenicity test and K-B disc method, the isolated strain was identified as pathogenic E. coli .[Results] The isolated pathogen was highly sensitive to ceftriaxone, cefotaxime, ciprofloxacin and lincomycin, moderately sensitive to enrofloxacin, neomycin, gentamycin, spectinomycin, florfenicol, amikacin and polymyxin, and resistant to ampicillin, amoxicillin and doxycycline.[Conclusions] This study provided reference for the prevention and control of diarrheal diseases in foxes in Qinhuangdao region.展开更多
基金supported by Pancosma SA,Geneva,Switzerland,Jastro & Shields Graduate Research Awardthe United States Department of Agriculture (USDA) National Institute of Food and Agriculture (NIFA),multistate projects W4002 and NC1202
文摘Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to carbadox.The objective of this study was to investigate the impacts of oligosaccharide-based polymer or antibiotic on the host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.Results Multivariate analysis highlighted the differences in the metabolic profiles of serum and colon digesta which were predominantly found between pigs supplemented with oligosaccharide-based polymer and antibiotic.The relative abundance of metabolic markers of immune responses and nutrient metabolisms,such as amino acids and carbohydrates,were significantly differentiated between the oligosaccharide-based polymer and antibiotic groups(q<0.2 and fold change>2.0).In addition,pigs in antibiotic had a reduced(P<0.05)relative abundance of Lachnospiraceae and Lactobacillaceae,whereas had greater(P<0.05)Clostridiaceae and Streptococcaceae in the colon digesta on d 11 post-inoculation(PI)compared with d 5 PI.Conclusions The impact of oligosaccharide-based polymer on the metabolic and microbial profiles of pigs is not fully understood,and further exploration is needed.However,current research suggest that various mechanisms are involved in the enhanced disease resistance and performance in ETEC-challenged pigs by supplementing this polymer.
文摘BACKGROUND:The virulent factors of Escherichia coli(E.coli) play an important role in the process of pathopoiesis.The study aimed to compare drug-resistant genes and virulence genes between extended spectrum β-lactamases(ESBLs)-producing E.coli and non-ESBLs-producing E.coli to provide a reference for physicians in management of hospital infection.METHODS: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,qacEA1-sull and 8 virulence genes(CNF2,hlyA,eaeA,VT1,est,bfpA,elt,and CNF1) were tested by polymerase chain reaction(PCR).RESULTS:Among the 96 E.coli isolates,the ESBLs-producing E.coli comprised 46(47.9%)strains and the non-ESBLs-producing E.coli consisted of 50(52.1%) strains.The detection rates of multiple drug-resistant strain,qacEA1-sull,CNF2,hlyA,eaeA,VT1,est,bfpA,elt,and CNF1 in 46ESBLs-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.coli strains,the positive rates of multiple drug-resistant strain,qacEA1-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.coli strains and the non-ESBLs-producing E.coli strains was statistically significant(P<0.05).CONCLUSION:The positive rate of multiple drug-resistant strains is higher in the ESBLsproducing 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.
文摘Background: Recently micro-organisms that synthesize extended-spectrum β-lactamase (ESBLs) were increased. The peculiarities of ESBL synthesis of Escherichia coli and Klebsiella pneumoniae strains that cause nosocomial urinary tract infections, surgical site infections and pneumonia in surgical clinic were studied. ESBL synthesis were observed 38.9% of E. coli strains obtained from urine, 92.3% of strains obtained from surgical site infections, and 50% of strains obtained from sputum. ESBL synthesis were observed 37.5% of K. pneumoniae strains obtained from urine, 85.7% of strains obtained from surgical site infections, and 60% of strains obtained from sputum. Different levels of ESBL synthesize of E. coli and K. pneumoniae strains isolated from different pattern is discussed. Conclusion. ESBL synthesis is common in E. coli and K. pneumoniae strains, which cause nosocomial infections. The frequency of occurrence of ESBL s synthesis among of these strains depends on clinical forms of nosocomial infections.
文摘Fecal coliform bacteria such as Escherichia coli (E. coli) are one of the main sources of groundwater pollution. An assessment of the transport and Persistence of E. coli in poultry litter amended Decatur silty Clay soil and Hartsells Sandy soil was conducted using soil columns and simulated groundwater leaching. Enumeration of initial E. coli was determined to range from 2.851 × 10<sup>3</sup> to 3.044 × 10<sup>3</sup> CFU per gram of soil. These results have been used in a batch study to determine the persistence rate of E. coli in Decatur silty Clay soil and Hartsells Sandy soil. Results prove that E. coli survival growth rate increases for clay soil later than and at a higher rate than sandy soil. The column study has determined that E. coli was transported at a rate of 3.7 × 10<sup>6</sup><sup> </sup>CFU for Decatur silty loam and 6.3 × 10<sup>6</sup><sup> </sup>CFU for Hartsells sandy per gram of soil. Further, linear regression analysis predictions show higher porosity and soil moisture content affect transport, and Hartsells sandy soil has higher transport of E. coli due to its higher porosity and lower volumetric water content.
基金supported by the National Natural Science Foundation of China(Grant No.50671101).
文摘A preliminary study was made on the antibacterial mechanism of copper-bearing antibacterial stainless steels against E.coli through experiments of microbiology such as EDTA (ethylenediaminetetraacetic acid) complexing, DNA smearing and AFM (atomic force microscope) observation. It was measured that the antibacterial stainless steels showed excellent antibacterial functions with antibacterial rate to E.coli over 99.99%. The antibacterial rate was weak if the bacteria solution was complexed by EDTA, indicating that the copper ions play a dominant role in the antibacterial effect of the antibacterial stainless steels. The electrophoresis experiment did not show the phenomenon of DNA smearing for E.coli after contacting antibacterial stainless steels, which meant that DNA of E.coli was not obviously damaged. It was observed by AFM that the morphology of E.coli changed a lot after contacting antibacterial stainless steels, such as cell walls being seriously changed and lots of contents in the cells being leaked.
基金Funding for this project was provided by Alberta Agriculture and Forestry Strategic Research and Development Section,Evonik Nutrition&Care GmbH,and Mitacs Accelerate.
文摘Background:The independent and interactive effects of dietary fiber(DF)and threonine(Thr)were investigated in growing pigs challenged with either systemic E.coli lipopolysaccharide(LPS)or enteric Salmonella Typhimurium(ST)to characterise their effect on intestinal barrier function.Results:In experiment 1,intestinal barrier function was assessed via oral lactulose and mannitol(L:M)gavage and fecal mucin analysis in pigs challenged with E.coli LPS and fed low fiber(LF)or high fiber(HF)diets with graded dietary Thr.Urinary lactulose recovery and L:M ratio increased(P<0.05)during the LPS inoculation period in LF fed pigs but not in HF fed pigs.Fecal mucin output was increased(P<0.05)in pigs fed HF compared to LF fed pigs.In experiment 2,RT-qPCR,ileal morphology,digesta volatile fatty acid(VFA)content,and fecal mucin output were measured in Salmonella Typhimurium challenged pigs,fed LF or HF diets with standard or supplemented dietary Thr.Salmonella inoculation increased(P<0.05)fecal mucin output compared to the unchallenged period.Supplemental Thr increased fecal mucin output in the HF-fed pigs(Fib×Thr;P<0.05).Feeding HF increased(P<0.05)VFA concentration in cecum and colon.No effect of either Thr or fiber on expression of gene markers was observed except a tendency(P=0.06)for increased MUC2 expression with the HF diet.Feeding HF increased goblet cell numbers(P<0.05).Conclusion:Dietary fiber appears to improve barrier function through increased mucin production capacity(i.e.,goblet cell numbers,MUC2 gene expression)and secretion(i.e.,fecal mucin output).The lack of effect of dietary Thr in Salmonella-challenged pigs provides further evidence that mucin secretion in the gut is conserved and,therefore,Thr may be limiting for growth under conditions of increased mucin production.
基金This work was supported by National Natural Science Foundation of China 30070038 and 30130070, National High-Tech Research and Development Program of China (863 Project) 2001AA215391, and EU Project QLRT 2000 01441.
文摘Objective To report a protocol using biotin-labelled PrP protein in cell free conversion assay instead of isotope. Methods A hamster PrP protein (HaPrP) was expressed in E. coli and purified with HIS-tag affinity chromatograph. After being labelled with biotin, HaPrP was mixed with PrP^sen preparation from scrapie strain 263K. Results Protease-resistant bands were detected after four-day incubation. Conclusion The new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.
基金Supported by the National Natural Science Foundation of China,No.30070020
文摘AIM: To study the effect of integration of tandem aroG-pheA genes into the tyrA locus of Corynebacterium glutamicum (C. glutamicum) on the production of L-phenylalanine.METHODS: By nitrosoguanidine mutagenesis, five pfluorophenylalanine (FP)-resistant mutants of Cglutamicum FP were selected. The tyrA gene encoding prephenate dehydrogenase (PDH) of C.glutamicum was amplified by polymerase chain reaction (PCR) and cloned on the plasmid pPR. Kanamycin resistance gene (Km) and the PBF-aroGpheA-T (GA) fragment of pGA were inserted into tyrA gene to form targeting vectors pl-K and pTGAK, respectively. Then,they were transformed into C.glutamicum FP respectively by electroporation. Cultures were screened by a medium containing kanamycin and detected by PCR and phenotype analysis. The transformed strains were used for L-phenylalanine fermentation and enzyme assays.RESULTS: Engineering strains of C.glutamicum (Tyr-)were obtained. Compared with the original strain, the transformed strain C. glutamicum GAK was observed to have the highest elevation of L-phenylalanine production by a 1.71-fold,and 2.9-, 3.36-, and 3.0-fold in enzyme activities of chorismate mutase, prephenate dehydratase and 3-deoxy-Darabinoheptulosonate-7-phosphate synthase, respectively.CONCLUSION: Integration of tandem aroG-pheA genes into tyrA locus of C. glutamicum chromosome can disrupt tyrA gene and increase the yield of L-phenylalanine production.
基金support by Australian Cooperative Research Centre for High Integrity Australian Pork(Award number 2C-110 1213)
文摘Background: This experiment was conducted to test the hypothesis that vitamin E(Vit E) and acetylsalicylic acid(ASA), a cyclooxygenase-2(COX-2) inhibitor, will additively reduce the production of the immunosuppressive molecule prostaglandin E_2(PGE_2) and hence reduce inflammatory responses in weaner pigs experimentally infected with an enterotoxigenic strain of E. coli.Methods: The experiment was conducted in a research facility with 192 individually-housed male weaner pigs(Landrace × Large White) weighing 6.6 ± 0.04 kg(mean ± SEM). The pigs were experimentally infected with an enterotoxigenic strain of E. coli and were allocated to a 2 × 3 factorial design with the respective factors being without and with 125 ppm ASA and three levels of Vit E supplementation(50, 100 or 200 IU/kg diet, dl-α-tocopheryl acetate).Results: Acetylsalicylic acid supplementation improved average daily gain(P 〈 0.05) and tended to improve feed:gain ratio(P 〈 0.10) during the first 14 d after weaning. Acetylsalicylic acid supplementation also improved(P 〈 0.001) amino acid utilization efficiency(as assessed by plasma urea level) and tended to decrease(P 〈 0.10) PGE2 production in the liver without affecting smal intestinal histology and tight junction protein mR NA expression in the jejunal epithelium. Vitamin E supplementation greater than 100 IU/kg diet sustained both the plasma Vit E concentration(P 〈 0.001) and plasma haptoglobin content(P 〈 0.001) after weaning. However, there was no additive effects of the combined supplementation of ASA and Vit E on performance, intestinal barrier function and inflammatory responses of weaned pigs.Conclusions: Although ASA and vitamin E improved amino acid utilization efficiency and reduced acute inflammatory responses, ASA and vitamin E did not additively reduce production of PGE2 and inflammatory responses in weaner pigs experimental y infected with an enterotoxigenic strain of E. coli.
文摘Objective To investigate whether the effect of E.coli on U937 cell lines apoptosis is mediated via p38 mitogen-activated protein kinase (MAPK) activation. Methods The U937 cell lines were treated with E.coli at different time or together with SB203580, an inhibitor for p38. Cell apoptosis was analyzed by flow cytometry. p38 activities were detected by Western blotting. Results E.coli induced apoptosis in cultured U937 cell lines in a time-dependent manner. The phosphorylation of p38 was induced after 10 minutes infection, reached the peak after 20 minutes, and started to decline after 30 minutes. In contrast, the level of total p38 protein was not changed in whole experimental period. Inhibition of p38 with SB203580 significantly inhibited E.coli induced apoptosis in U937 cells. Conclusion The activation of the p38 MAPK in U937 cell lines by E.coli is a major pathway to mediate the apoptosis.
文摘Ciprofloxacin is a second-generation of fluoroquinolone,broad-spectrum antibiotic with bactericidal activity against Gram-positive and Gram-negative organisms.It is one of the most widely used antibiotics,because of its efficacy,safety,and relatively low cost.Ascorbic acid(vitamin C)is water-soluble monosaccharide antioxidant;it is essentially required by the body for its various biochemical and physiological processes.S.aureus is Gram-positive cocci;widely distributed in the environment,it is a member of the normal flora of the body.S.aureus is not always pathogenic;it is a common cause of skin infections including abscesses,respiratory infections such as sinusitis,and food poisoning.E.coli is Gram-negative bacteria,found in the environment,foods,and intestines.Most E.coli strains are harmless;it is part of the normal microbiota of the gut.However,some serotypes of E.coli cause serious food poisoning in their hosts;it can cause diarrhea,while others cause urinary tract infections,respiratory illness and pneumonia,and other illnesses.Method:Cup cut diffusion method was applied.Experiment I:is carried out to choose the concentration of vitamin C to be used in experiment II.The negative control is normal saline,added in cup in each plate,vitamin C(100 mg/mL,200 mg/mL,400 mg/mL)was added,the volume in each cup was 100μL.Experiment II:Eight groups of treatments were applied.The first is the negative control(1%normal saline),the second group is the positive control of vitamin C(200 mg/mL).The third,fourth and fifth groups are ciprofloxacin with different concentrations(10 mg/mL,20 mg/mL,40 mg/mL);the sixth,seventh and eighth are the combination of vitamin C with each concentration of ciprofloxacin(10 mg/mL,20 mg/mL,40 mg/mL).Each group includes six petri dishes.Bacterial plates were incubated at 37 oC for 24 h and 48 h.Zone of inhibition is measured in mm.Results and conclusion:Ciprofloxacin produces dose dependent increase in zone of inhibition of S.aureus and E.coli growth,after 24 and 48 hours incubation.While vitamin C in the concentration used produced inhibitory effect on the growth of S.aureus and E.coli,after 24 hours incubation,vitamin C effect was not changed after 48 hours incubation.After 24 hours incubation,vitamin C potentiated the effect of ciprofloxacin at low concentration(10 mg/mL);while vitamin C antagonized the effect of ciprofloxacin at higher concentrations(20 and 40 mg/mL)on S.aureus growth.In the same time,ciprofloxacin antagonized the inhibitory effect of vitamin C on S.aureus growth.After 48 hours incubation,S.aureus produced resistance against ciprofloxacin alone,and that combined with vitamin C compared to zone of inhibition after 24 hours.Ciprofloxacin produced dose dependent inhibition of E.coli growth after incubation for 24 and 48 hours.Vitamin C potentiated the inhibitory effect induced by ciprofloxacin(additive effect).The inhibitory effect of ciprofloxacin,vitamin C and the combination was not changed after 48 hours compared to 24 hours.
基金supported by 2009--0214 numbered project of National Boron Research Institute(BOREN)-Turkey
文摘Objective To study MIC value of 7 boron derivatives namely [Boric acid (H3BO3), Anhydrous Borax (Na2B407), Sodium Borate (NaBO2), Diammonium Tetraborate (NH4)2B4O7, Sodium Perborate (NaBO3), Boron Trioxide (B203), Potassium Tetraborate (K2B407)] on E. coil and P. aeruginosa and their effects on survival of bacteria in lake water and resistance against kanamycin antibiotic. Methods MIC values of Boron derivatives and antibiotic were studied by broth microdilution method. The effect of boron derivatives on survival of bacteria in lake water were also determined with plate count. Results Sodium perborate was determined as the substances. Effectiveness increased as temperature most effective substance among the studied increased. E. coil was more affected from P. aeruginosa in 8 mg/mL sodium perborate concentration in lake water. Moreover, it was determined that MIC value of kanamycin antibiotic decreased 200 times by especially treating P. aeruginosa with sodium perborate in lake water. However, it can be stated that this change in resistance did not arise from microorganisms. Conclusion Sodium perborate solution can be used supportedly in kanamycin antibiotic applications for P. aeruginosa. Future studies are necessary to explore the relation between sodium perborate and kanamycin which is effective on P. aeruginosa in lake water.
基金Supported by the National Natural Science Foundation of China (30170213) and the Foundation of Wuhan University(0000028)
文摘The Ca. LSU intron flanking a 129 bp cxon upstream and a fOO bp exondownstream was inserted into the lacZ gene on pRS426 to transform E. coli. Northern blot analysisand RT-PCR showed that splicing of Ca. LSU in E. coli is efficient upon inducible expression of theprecursor RNA, In contrast, co-lranscriptional self-splicing of the intron in vitro is much lessactive. Therefore, this E. coli splicing system can be used as a better model to investigate theeffect of the ribozyme inhibitors on Ca. LSUsplicing in living cell. We examined (he effects ofneomycin sulfatc and pentamidine on Ca. LSU splicing in E. coli, and found that these drugsdoes-dependently inhibit the intron splicing. However, heomycin is more potent than pentamidine inthis action.
基金This study was supported by the Science & Technology Plan (No. 2001C12001) of Guangdong Province,P.R. China
文摘To obtain the recombinant core domain of porcine zone pellucida 3β (cZP3β) for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained by PCR and then was cloned into pET-3c vector. After being identified, recon was transformed into E.coli BL21 (DE3) pLysS and then induced by IPTG. Results The recombinant cZP3β was expressed in E. coli up to 15% of total cellular proteins, and was made sure by Western blot analysis. Conclusion The research on expression of core domain of pZP3β could benefit to further investigation of its immunogenicity and the development of antigen preparation.
基金Funded by the National Natural Science Foundation of China (No. 50702040)
文摘The photocatalytic degradation of E.coli membrane cell by ZnO nanowires was studied using field-emission scanning electron microscope(FE-SEM),fluorescence microscopy,and Attenuated total reflection fourier transform infrared(ATR-FTIR).The outer membrane of E.coli was removed completely in the presence of ZnO nanowires under UV irradiation,and the cells became twisted shapes without a mechanically strong network.After ZnO nanowires photocatalysis,the permeability of the treated cells increased to some degree that could be confirmed by quantum dots labeling technique.Structural changes in the cell wall membrane were revealed by the decay of the characteristic groups bands in ATR-FTIR spectra.
基金The autors are thankful to Kurdistan Digestive,Liver Research Center which is affiliated to Kurdistan University of Medical Sciences,Sanandaj,for providing the financial support and necessary facilities
文摘Objective:Diarrhea is a leading cause of morbidity and mortality among children in developing countries.The bacterial pathogen most commonly associated with childhood diarrhea is Escherichia coli.A one-year prospective study was carried out in Sanandaj to determine the prevalence and roles of the different E.coli pathotypes in children less than five years of age with acute diarrhea.Methods:Rectal swab were collected prospectively from children with acute diarrhea and transported to the Department of Microbiology,School of Medicine, KUMS,Sanandaj during 2008.The study was approved by the institutional ethics committee.Results:During this study period,rectal swabs were investigated from a total of 466 children 1 to 144 months of age(mean, 29.97 months±S.D) with diarrhea.Among the children,191(41%,191/466) were girls,and 275 (59%,275/466) were boys.The age-specific incidence rates of acute diarrhea among children 13-24 and 1 - 12 months of age were 37.37%(37/99) and 26.26%(26/99),respectively,during the study period.A total of 99 strains of E.coli were detected.EPEC 59(59.59%) and EIEC 22(22.22%),were the most commonly found Escherichia coli strains detected in stools from children.Disk diffusion testing showed E.coli strains resistance to tetracycline(89.89%),chloramphenicol(88.88%),Ampicillin(79.79%),Amoxicillin (75.75%) and Ceficime(75.75%).Among risk factors like age,sex,haemoglubin,fathers and mothers education,food and weight of children only mother's education was significant(P =0.018).Conclusion: In most of the clinical laboratories in Iran,E.coli does not considered as an etiologic agent responsible for diarrhea. Results in this study revealed that E.coli should be considered as an etiologic agent causing acute diarrhea among children.We therefore,recommend the routine isolation and identification of E.coli strains in all the clinical laboratories in Sanandaj.Guidelines for appropriate use of antibiotics in Sanandaj need updating.
基金Supported by Project of Hebei Education Department(ZD2017234)Project of Science and Technology Bureau of Shijiazhuang(171500953A)Project of Science and Technology Bureau of Qinhuangdao(201602A046)
文摘[Objectives] The study aimed to identify the pathogenic E. coli strain that caused diarrhea in foxes and to analyze its drug sensitivity.[Methods] A pathogenic E. coli strain was isolated from dead foxes with diarrhea. By conventional bacterial isolation and culture, morphological observation, pathogenicity test and K-B disc method, the isolated strain was identified as pathogenic E. coli .[Results] The isolated pathogen was highly sensitive to ceftriaxone, cefotaxime, ciprofloxacin and lincomycin, moderately sensitive to enrofloxacin, neomycin, gentamycin, spectinomycin, florfenicol, amikacin and polymyxin, and resistant to ampicillin, amoxicillin and doxycycline.[Conclusions] This study provided reference for the prevention and control of diarrheal diseases in foxes in Qinhuangdao region.