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the escherichia coli o157:h7 ehab autotransporter protein binds to laminin and collagen i and induces a serum iga response in o157:h7 challenged cattle.enterohaemorrhagic escherichia coli (ehec) are a subgroup of shiga toxin-producing e.  coli that cause gastrointestinal disease with the potential for life-threatening sequelae. cattle serve as the natural reservoir for ehec and outbreaks occur sporadically as a result of contaminated beef and other farming products. while certain ehec virulence mechanisms have been extensively studied, the factors that mediate host colonization are poorly defined. previously, we identified four proteins (ehaa,b ...200919508554
quarter and cow risk factors associated with the occurrence of clinical mastitis in dairy cows in the united kingdom.quarter and cow risk factors associated with the development of clinical mastitis (cm) during lactation were investigated during a 12-mo longitudinal study on 8 commercial holstein-friesian dairy farms in the southwest of england. the individual risk factors studied on 1,677 cows included assessments of udder and leg hygiene, teat-end callosity, and hyperkeratosis; body condition score; and measurements of monthly milk quality and yield. several outcome variables for cm were used for statistical ...200919447987
separation of acidic and basic proteins by ce with ctab additive and its applications in peptide and protein profiling.in this paper, we report a very simple but efficient cationic micellar ce method for protein analysis in complex samples without any modification for the capillary wall. it was found that increased concentration (>or=2 mm) of ctab additive in electrophoretic buffer can effectively suppress the wall adsorption of both acidic and basic proteins. the separation was improved with buffer ph decreasing from 6.0 to 3.0. by using a buffer containing 2 mm ctab at ph 3.0, nine proteins with a wide range o ...200919582714
evidence-based semiquantitative methodology for prioritization of foodborne zoonoses.to prioritize an extended list of food- and water-borne zoonoses to allow food safety authorities to focus on the most relevant hazards in the food chain.200919715429
virulence gene and antibiotic resistance profile of shiga-toxin-producing escherichia coli prevalent in captive yaks (poephagus grunniens). 200919477084
[expression of the genes cela and xyla isolated from a fragment of metagenomic dna in escherichia coli].the glycosyl hydrolase genes cel5a and xyl3a previously isolated by ourselves within a fragment of dna from the methagenomic library of cow rumen microflora dna were sub-cloned and expressed in e. coli. the recombinant proteins cel5a and xyl3a were purified and characterized. cellulase cel5a belongs to the family 5 glycosyl hydrolases and is a one-module 38.2 kda enzyme that hydrolyses the 1,4-glycoside bonds of soluble cellulose substrates and amorphous cellulose, showing its maximal activity ( ...200919517808
inhibition of uropathogens by lactic acid bacteria isolated from dairy foods and cow's intestine in western nigeria.a total of 96 lactic acid bacteria (lab) were isolated from african indigenous fermented products and cow's intestines to study their inhibitory capability against multi-drug-resistant uropathogens. escherichia coli accounted for approximately 45% of isolated uropathogens, followed by staphylococcus spp. (20%). the gram negative uropathogens were highly resistant to quinolones, co-trimoxazole, teicoplanin and some beta-lactams, while the staphylococcus spp. showed high resistance to aminoglycosi ...200919529917
recombinant expression and affinity purification of a novel epididymal human sperm-binding protein, bsph1.mammalian sperm undergo a series of maturation steps before acquiring fertilization competence. our previous work demonstrated the importance of binder of sperm (bsp) proteins in bovine sperm capacitation. recent studies identified a bsp-homologous dna sequence in the human genome (bsph1) and mrna expression in the epididymis. the aim of this study was to develop an efficient method to express and purify recombinant human bsph1. bsph1 accumulates in inclusion bodies when expressed with an n-term ...200919091820
arsenic(iii) species inhibit oxidative protein folding in vitro.the success of arsenic trioxide in the treatment of acute promyelocytic leukemia has renewed interest in the cellular targets of as(iii) species. the effects of arsenicals are usually attributed to their ability to bind vicinal thiols or thiol selenols in prefolded proteins thereby compromising cellular function. the present studies suggest an additional, more pleiotropic, contribution to the biological effects of arsenicals. as(iii) species, by avid coordination to the cysteine residues of unfo ...200919102631
pathogen prevalence and influence of composted dairy manure application on antimicrobial resistance profiles of commensal soil bacteria.composting manure, if done properly, should kill pathogenic bacteria such as salmonella and escherichia coli o157:h7, providing for an environmentally safe product. over a 3-year period, samples of composted dairy manure, representing 11 composting operations (two to six samples per producer; 100 total samples), were screened for salmonella and e. coli o157:h7 and were all culture negative. nonpathogenic bacteria were cultured from these compost samples that could theoretically facilitate the sp ...200919105635
transcriptome profiling of streptococcus uberis-induced mastitis reveals fundamental differences between immune gene expression in the mammary gland and in a primary cell culture model.streptococcus uberis is a prevalent causative organism of mastitis and resides naturally in the environment of the dairy cow making prevention of the disease difficult. a bovine cdna microarray comprising approximately 22,000 expressed sequence tags was used to evaluate the transcriptional changes that occur in the mammary gland after the onset of clinical strep. uberis mastitis. five lactating friesian heifers were intramammary infused in an uninfected quarter with approximately 1,000 to 1,500 ...200919109270
supplemental fish oil does not alter immune competence or the pathophysiological response to an intramammary infusion of endotoxin in peri-partum multiparous holstein cows.the objective was to determine the effects of supplementing the diet with fish oil during the peri-partum period on the immune competence and the pathophysiological response to a lipopolysaccharide-induced mastitis challenge. multiparous holstein cows (n=30) were completely randomized to one of two treatments at 3 weeks pre-partum. treatments differed only in the source of supplemental lipid and included either energy booster or fish oil. treatment diets were fed from -21 d relative to expected ...200919121237
defatted bovine colostrum-supplemented diet around weaning improves exocrine pancreatic secretion by means of volume, digestive enzymes, and antibacterial activity.exocrine pancreatic secretion contributes to limit pathogenic bacteria-associated diarrhea. bovine colostrum, used in the treatment of diarrhea, reduces symptoms originating from gut pathogenic bacteria overgrowth. we hypothesized that bovine colostrum may stimulate the exocrine pancreatic secretion.200919136910
biochemical analysis of a beta-d-xylosidase and a bifunctional xylanase-ferulic acid esterase from a xylanolytic gene cluster in prevotella ruminicola 23.prevotella ruminicola 23 is an obligate anaerobic bacterium in the phylum bacteroidetes that contributes to hemicellulose utilization within the bovine rumen. to gain insight into the cellular machinery that this organism elaborates to degrade the hemicellulosic polymer xylan, we identified and cloned a gene predicted to encode a bifunctional xylanase-ferulic acid esterase (xyn10d-fae1a) and expressed the recombinant protein in escherichia coli. biochemical analysis of purified xyn10d-fae1a reve ...200919304844
establishing relative release kinetics of faecal indicator organisms from different faecal matrices.a laboratory assay for comparative characterization of various faecal matrices with respect to faecal indicator organism (fio) release using, artificial rain water.200919422475
pulsed-field gel electrophoresis profile changes resulting from spontaneous chromosomal deletions in enterohemorrhagic escherichia coli o157:h7 during passage in cattle.a total of 905 enterohemorrhagic escherichia coli (ehec) o157:h7 isolates that were recovered from experimentally infected cattle, in addition to the inoculated strain, were analyzed by pulsed-field gel electrophoresis (pfge). twelve pfge profiles other than that of the inoculated strain were observed. we successfully identified five distinct chromosomal deletions that affected the pfge profiles using whole-genome pcr scanning and dna sequencing analysis. the changes in pfge profiles of ehec o15 ...200919581472
fluoresceination of fepa during colicin b killing: effects of temperature, toxin and tonb.we studied the reactivity of 35 genetically engineered cys sulphydryl groups at different locations in escherichia coli fepa. modification of surface loop residues by fluorescein maleimide (fm) was strongly temperature-dependent in vivo, whereas reactivity at other sites was much less affected. control reactions with bovine serum albumin showed that the temperature dependence of loop residue reactivity was unusually high, indicating that conformational changes in multiple loops (l2, l3, l4, l5, ...200919432807
defining postpartum uterine disease and the mechanisms of infection and immunity in the female reproductive tract in cattle.uterine microbial disease affects half of all dairy cattle after parturition, causing infertility by disrupting uterine and ovarian function. infection with escherichia coli, arcanobacterium pyogenes, and bovine herpesvirus 4 causes endometrial tissue damage. toll-like receptors on endometrial cells detect pathogen-associated molecules such as bacterial dna, lipids, and lipopolysaccharide (lps), leading to secretion of cytokines, chemokines, and antimicrobial peptides. chemokines attract neutrop ...200919439727
absence of escherichia coli phylogenetic group b2 strains in humans and domesticated animals from jeonnam province, republic of korea.multiplex pcr analyses of dnas from genotypically unique escherichia coli strains isolated from the feces of 138 humans and 376 domesticated animals from jeonnam province, south korea, performed using primers specific for the chua and yjaa genes and an unknown dna fragment, tspe4.c2, indicated that none of the strains belonged to e. coli phylogenetic group b2. in contrast, phylogenetic group b2 strains were detected in about 17% (8 of 48) of isolates from feces of 24 wild geese and in 3% (3 of 9 ...200919592524
[cloning, prokaryotic expression of cattle ghrelin gene and biological activity detection of the expressed protein].the cdna of cattle ghrelin gene was amplified from abomasum fundic gland mrna of qinchuan cattle by rt-pcr. pcr product was cloned into the t vector pgm-t to construct pgh-t1 for sequencing. then the cdna was subcloned into the prokaryotic expressing plasmid vector pet32a (+) and transformed into host escherichia coli strain bl21 (de3) for expression. the expression of pgh-32 mature ghrelin protein was induced by iptg and was identified by sds-page. the expression product was observed with solub ...200919441222
adjuvant effect of bovine heat shock protein 70 on piroplasm surface protein, p33, of theileria sergenti.in this study, we investigated the immunological effects of bovine heat shock protein 70 (hsp70) on the major theileria sergenti surface protein (p33). the gene encoding p33 was expressed as a fusion protein with bovine hsp70 from a plasmid vector. the adjuvant function of hsp70 on p33 was evaluated with regard to antibody response, cytokine production, and a challenge experiment in mice or cattle. hsp-p33 fusion protein provoked higher humoral and cellular immunity than either escherichia coli- ...200919443238
influence of body condition and forage type on prevalence of escherichia coli o157:h7 and salmonella in grazing beef cows.to determine the influence of body condition (bc) and forage type on the prevalence of faecal shedding of escherichia coli o157:h7 and salmonella from beef cows.200919627482
cell population, viability, and some key immunomodulatory molecules in different milk somatic cell samples in dairy cows.immune cells in the milk are most important in combating pathogens that invade the mammary gland. this study investigated the immune competence and viability of somatic milk cells that are already resident in milk and udders free of infection. cells were studied in freshly removed milk to simulate conditions in the udder. effects of incubation, cell preparation, and immunological stimulation with 0.5 mug/ml lipopolysaccharide (lps) from escherichia coli were analysed. viability and differential ...200919445829
rapid quantification of salmonella typhimurium inoculated to meat products by real-time pcr.the objective of this study was to use a 5'-nuclease (taqman) real-time pcr method with primers and probe specific to the spaq gene as a rapid approach to quantitatively determine salmonella typhimurium. the result showed that the correlation coefficient between real-time pcr estimates and bovine serum albumin (bsa) plate counts of s. typhimurium was 0.99, independently of 10(5)-fold numbers of bystander escherichia coli o157:h7 or total viable counts. the sensitivity of the real-time quantitati ...200919457771
a biotin-hydrogel-coated quartz crystal microbalance biosensor and applications in immunoassay and peptide-displaying cell detection.a biotin-coated quartz crystal microbalance (qcm) chip was prepared by dip-coating a long-chain alkanethiol-modified crystal with precoupled dextran-biotin hydrogels. the resulting biotin chip was used to affinity-immobilize streptavidin (sav) and was then further employed for various biosensor assays. first, the sav chip allowed efficient on-line binding of biotinylated bovine serum albumin (bbsa), followed by a sensitive and specific response toward anti-bovine serum albumin (bsa) antibodies. ...200919464992
characterisation of atypical enteropathogenic e. coli strains of clinical origin.enteropathogenic e. coli (epec) is a prominent cause of diarrhoea, and is characterised in part by its carriage of a pathogenicity island: the locus for enterocyte effacement (lee). epec is divided into two subtypes according to the presence of bundle-forming pili (bfp), a fimbrial adhesin that is a virulence determinant of typical epec (tepec), but is absent from atypical epec (aepec). because aepec lack bfp, their virulence has been questioned, as they may represent lee-positive shiga toxin-pr ...200919490652
genotypic diversity of escherichia coli in a dairy farm.dairy cattle are known reservoirs of pathogenic escherichia coli, but little is known about the dynamics of e. coli in dairy cows or within the dairy farm environment. this study was conducted to evaluate the diversity and distribution of e. coli strains in a dairy farm using pulsed-field gel electrophoresis and to determine the relationships between e. coli isolated from feces and throughout the farm environment. water from watering troughs, feces from cows, manure composites, milk, and milk fi ...200919459756
antimicrobial activity of bovine psoriasin.human psoriasin (s100a7) has originally been described as a member of the family of s100 calcium-binding proteins which is overexpressed in patients suffering from psoriasis. the bovine homolog was first identified as a cow-derived respiratory allergen. as escherichia coli mastitis is a common problem in dairy cattle, and human psoriasin was found to exhibit antimicrobial activity preferentially against e. coli, we examined whether the bovine mrna is expressed in the mammary gland. to demonstrat ...200919167844
characterization of the alpha-haemolysin determinant from the human enteropathogenic escherichia coli o26 plasmid peo5.the 157-kb conjugative plasmid peo5 encoding alpha-haemolysin in strains of human enteropathogenic escherichia coli (epec) o26 was investigated for its relationship with ehec-haemolysin-encoding plasmids of enterohaemorrhagic e. coli (ehec) o26 and o157 strains. plasmid peo5 was found to be compatible with ehec-virulence plasmids and did not hybridize in southern blots with plasmid po157 from the ehec o157:h7 strain edl933, indicating that both plasmids were unrelated. a 9227-bp stretch of peo5 ...200919175679
effects of the dicarboxylic acids malate and fumarate on e. coli o157:h7 and salmonella enterica typhimurium populations in pure culture and in mixed ruminal microorganism fermentations.the dicarboxylic acids malate and fumarate increase ruminal ph, reduce methane production, increase propionate and total volatile fatty acid (vfa) production, and reduce lactic acid accumulation in a manner similar to ionophores. these acids stimulate the ruminal bacterium selenomonas ruminantium to ferment lactate to produce propionate. thus, dicarboxylic acids have been suggested as nonantibiotic modifiers of the ruminal fermentation, but their impact on ruminal microbial ecology remains unkno ...200919194750
a biosecure composting system for disposal of cattle carcasses and manure following infectious disease outbreak.during outbreaks of infectious animal diseases, composting may be an effective method of disposing of mortalities and potentially contaminated manure. duplicate biosecure structures containing 16 cattle (bos taurus) mortalities (343 kg average weight) were constructed with carcasses placed on a 40-cm straw layer and overlaid with 160 cm of feedlot manure. at a depth of 80 cm (p80), compost heated rapidly, exceeding 55 degrees c after 8 d and maintained temperatures of 55 to 65 degrees c for > 35 ...200919202014
diversity in cell properties and transport behavior among 12 different environmental escherichia coli isolates.escherichia coli is a commonly used indicator organism for detecting the presence of fecal-borne pathogenic microorganisms in water supplies. the importance of e. coli as an indicator organism has led to numerous studies looking at cell properties and transport behavior of this microorganism. in many of these studies, however, only a single strain of e. coli was used even though research has shown that significant genetic variability exists among different strains of e. coli. if this genetic div ...200919202016
automated panning and screening procedure on microplates for antibody generation from phage display libraries.antibody phage display technology is well established and widely used for selecting specific antibodies against desired targets. using conventional manual methods, it is laborious to perform multiple selections with different antigens simultaneously. furthermore, manual screening of the positive clones requires much effort. the authors describe optimized and automated procedures of these processes using a magnetic bead processor for the selection and a robotic station for the screening step. bot ...200919224869
quantifying exposure to vero-cytotoxigenic escherichia coli o157 in milk sold as pasteurized: a model-based approach.milk sold as pasteurized has historically been implicated in the uk and worldwide as a vehicle for outbreaks of food-borne gastrointestinal disease, with a number of causative pathogenic organisms. one such organism is verocytotoxigenic escherichia coli, or e. coli, o157 (vtec o157). we present a quantitative assessment of likely exposure to vtec o157 via milk sold as pasteurized in the uk. particular interest in our assessment concerns whether there is any differential risk between milk which i ...200919232769
cytokine and acute phase protein gene expression in repeated liver biopsies of dairy cows with a lipopolysaccharide-induced mastitis.a minimally invasive liver biopsy technique was tested for its applicability to study the hepatic acute phase response (apr) in dairy cows with escherichia coli lipopolysaccharide (lps)-induced mastitis. the hepatic mrna expression profiles of the inflammatory cytokines, tumor necrosis factor alpha (tnf-alpha), il-1beta, il-6, and il-10, and the acute phase proteins serum amyloid a isoform 3 (saa3), haptoglobin (hp), and alpha(1)-acid glycoprotein (agp) were determined by real-time reverse trans ...200919233785
analytical specificity and sensitivity of a real-time polymerase chain reaction assay for identification of bovine mastitis pathogens.intramammary infection (imi), also known as mastitis, is the most frequently occurring and economically the most important infectious disease in dairy cattle. this study provides a validation of the analytical specificity and sensitivity of a real-time pcr-based assay that identifies 11 major pathogen species or species groups responsible for imi, and a gene coding for staphylococcal beta-lactamase production (penicillin resistance). altogether, 643 culture isolates originating from clinical bov ...200919233788
seasonal variation of shiga toxin-encoding genes (stx) and detection of e. coli o157 in dairy cattle from argentina.to study the seasonal variation of shiga toxin-encoding genes (stx) and to investigate the presence of shiga toxin-producing escherichia coli (stec) o157 in cattle belonging to five dairy farms from argentina.200919187162
a diva system based on the detection of antibodies to non-structural protein 3 (ns3) of bluetongue virus.vaccination programs for the control of bluetongue (bt) in ruminants have limitations due to difficulties in differentiating infected from vaccinated animals (diva). to overcome this problem a diva test that looks at a differential immune response to bluetongue virus (btv) non-structural protein 3 (ns3) was developed. the ns3 encoding gene of strain btv4/22045/pt04 was inserted into expression vector pet-28a and expressed in escherichia coli strain jm109. recombinant ns3 protein was used as an a ...200919233575
genotype, serotype, and antibiotic resistance of sorbitol-negative escherichia coli isolates from feedlot cattle.rectal fecal samples from 80 steers receiving rumensin, revalor-s, and liquamycin alone or in combination for growth promotion and disease prevention were examined for the presence of non-o157:h7 shiga toxin-producing escherichia coli. all isolates were identified with the api 20e test, virulence genes were detected with a pcr assay, and antibiotic susceptibilities were determined with the sensititre system. of the 153 e. coli isolates recovered 126 (82.3%) were sorbitol negative. isolates were ...200919205460
assessment of dietary ratios of red clover and grass silages on milk production and milk quality in dairy cows.twenty-four multiparous holstein-friesian dairy cows were used in a replicated 4 x 4 latin square changeover design experiment to test the effects of changing from ryegrass (lolium perenne) silage to red clover (trifolium pratense) silage in graded proportions on feed intakes, milk production, milk organoleptic qualities, and whole-body nitrogen partitioning. four dietary treatments, comprising ad libitum access to 1 of 4 forage mixtures plus a standard allowance of 4 kg/d dairy concentrates, we ...200919233807
phenotypic and genotypic traits of shiga toxin-negative e. coli o157:h7/h(-) bovine and porcine strains.enterohemorrhagic e. coli (ehec) o157:h7/h(-) (nonmotile) exist as shiga toxin gene (stx)-positive and stx-negative variants in patients and the environment. we analyzed molecular characteristics, phenotypes, and the phylogenetic background of three stx-negative e. coli o157:h7/h(-) strains isolated from cattle and a pig and compared them with those of human ehec and stx-negative e. coli o157:h7/h(-). all three animal strains contained flich7 and two contained eae. one eae-positive strain (o157: ...200919245340
selected antimicrobial resistance during composting of manure from cattle administered sub-therapeutic antimicrobials.composting is being increasingly employed for the recycling of nutrients in manure from the livestock industry. however, composting manure from animals fed antimicrobials has not been well characterized. in this study, compost windrows were prepared using manure collected from cattle (bos taurus l.) fed tylosin (ty), chlortetracycline-sulphamethazine (ts), and control cattle (no antimicrobials). the objectives of the 18-wk trial were to quantitatively assess the survival of total e. coli, e. col ...200919202027
effectiveness of 1,3-dibromo-5,5 dimethylhydantoin on reduction of escherichia coli o157:h7- and salmonella-inoculated fresh meat.1,3-dibromo-5,5-dimethylhydantoin (dbdmh; 25 degrees c) and hot water (85 degrees c) spray treatments were evaluated for efficacy in decontamination of pathogenic bacteria attached to beef carcass surfaces represented by cutaneous trunci (ct) muscle sections and beef hearts. treatments were evaluated using two different systems, a commercial carcass wash cabinet and a model carcass washer. the effects were measured immediately after treatment and again after 48 h of storage at 4 degrees c. secti ...200919205477
role of nitric oxide production in dairy cows naturally infected with mycobacterium avium subsp. paratuberculosis.nitric oxide (no) is a crucial mediator in host defense and is one of the major killing mechanisms within macrophages. its induction is highly affected by the types of cytokines and the infectious agents present. in the current study, no production was evaluated after in vitro infection of unfractionated peripheral blood mononuclear cells (pbmcs) with mycobacterium avium subsp. paratuberculosis (map) after 8h, 3 and 6 days of culture for cows in different stages of disease. in addition, the effe ...200919409621
effects of intramammary infusion of cis-urocanic acid on mastitis-associated inflammation and tissue injury in dairy cows.to evaluate the effects of cis-urocanic acid (cis-uca) on mammary gland (mg) inflammation and injury associated with escherichia coli-induced mastitis in dairy cows.200919254150
identification of escherichia coli and salmonella enterica organisms with reduced susceptibility to ceftriaxone from fecal samples of cows in dairy herds.to estimate the relationship between therapeutic use of ceftiofur and recovery of escherichia coli and salmonella spp with reduced susceptibility to ceftriaxone from feces of dairy cattle.200919254152
pentabody-mediated antigen delivery induces antigen-specific mucosal immune response.an efficient immunization system is essential for the development of mucosal vaccine. cholera toxin (ct) and escherichia coli heat labile toxin (lt) are among the strongest adjuvants tested in experimental animals but their use in humans has been hindered by their toxicity. on the other hand, the role of their non-toxic b-subunits, ctb or ltb, in enhancing mucosal immune response is not clear. we propose here a novel strategy for the induction of mucosal immune responses. single domain antibodie ...200919269688
sorbitol-fermenting shiga toxin-producing escherichia coli o157 in austria.infections with enterohemorrhagic escherichia coli (ehec) are the major cause of hemolytic uremic syndrome (hus), the most common cause of acute renal failure in childhood. shiga toxins are considered to be the most important virulence factor of ehec strains. non-sorbitol-fermenting ehec o157:h7 is still the most prevalent serotype isolated worldwide; however, sorbitol-fermenting (sf) ehec o157:h- (h- indicates nonmotility) strains are increasingly reported. thirteen sf ehec o157:h- strains (11 ...200919280135
the acute-phase protein serum amyloid a3 is expressed in the bovine mammary gland and plays a role in host defence.the serum amyloid a protein is one of the major reactants in the acute-phase response. using representational difference analysis comparing rna from normal and involuting quarters of a dairy cow mammary gland, we found an mrna encoding the saa3 protein (m-saa3). the m-saa3 mrna was localized to restricted populations of bovine mammary epithelial cells (mecs). it was expressed at a moderate level in late pregnancy, at a low level through lactation, was induced early in milk stasis, and expressed ...200919283521
protective antigen detection using horizontally stacked hexagonal zno platelets.anthrax toxin detection before bacteremia, when toxin concentration is low, improves the chances of efficient treatment and cure. we present a novel technique for ultrasensitive detection of a protective antigen (pa(83)) of anthrax using an array of zinc oxide nanorods in conjunction with a fitc-labeled pa affinity peptide. the nanorods are composed of horizontally stacked hexagonal platelets which are uniformly spaced and grown unidirectionally upon a glass substrate via a new and simple techni ...200919400578
evaluation of three commercially available enzyme-linked immunosorbent assay kits for detection of shiga toxin.three commercially available shiga toxin (stx) enzyme-linked immunosorbent assay (elisa) kits were evaluated for their ability to detect stx in pure cultures of stx-producing escherichia coli (specificity). the detection limits (sensitivity) of each elisa kit were also evaluated. seventy-eight stx-producing e. coli (stec) isolates that produced stx1, stx2, or stx1 and stx2 variants were examined in this study. the specificities of the tests were comparable, and the sensitivities of two of the te ...200919435221
global quantitative proteomic profiling through 18o-labeling in combination with ms/ms spectra analysis.several stable-isotope-based peptide labeling methods have been developed to support large-scale relative quantitation, through mass spectrometry, of proteins present in two different biological samples. in one of these, trypsin-catalyzed 18o-based labeling, quantitation is typically performed at the full scan (ms) level by comparing the peak intensities of sister precursor ions corresponding to the labeled and unlabeled forms of an intact peptide as they co-elute during liquid chromatography (l ...200919400582
mapping protein-protein interactions by localized oxidation: consequences of the reach of hydroxyl radical.hydroxyl radicals generated from a variety of methods, including not only synchrotron radiation but also fenton reactions involving chelated iron, have become an accepted macromolecular footprinting tool. hydroxyl radicals react with proteins via multiple mechanisms that lead to both polypeptide backbone cleavage events and side chain modifications (e.g., hydroxylation and carbonyl formation). the use of site-specifically tethered iron chelates can reveal protein-protein interactions, but the in ...200919354299
shiga toxin-producing escherichia coli o157 in beef and chicken burgers, and chicken carcasses in buenos aires, argentina.we describe the isolation and characterization of shiga toxin (stx)-producing escherichia coli (stec) o157:h7 from cooked and uncooked beef and chicken burgers and from chicken carcasses collected during sampling procedures in 2001 and 2002 in buenos aires city, argentina. of the 24 stec o157:h7 strains isolated, 20 were recovered from 19 (6.8%) out of 279 samples of beef and chicken burgers, and 4 strains from 4 (10.3%) out of 39 chicken carcasses. the samples were analyzed following the usda/f ...200919443067
maternally and naturally acquired antibodies to shiga toxins in a cohort of calves shedding shiga-toxigenic escherichia coli.calves become infected with shiga toxin-producing escherichia coli (stec) early in life, which frequently results in long-term shedding of the zoonotic pathogen. little is known about the animals' immunological status at the time of infection. we assessed the quantity and dynamics of maternal and acquired antibodies to shiga toxins (stx1 and stx2), the principal stec virulence factors, in a cohort of 27 calves. fecal and serum samples were taken repeatedly from birth until the 24th week of age. ...200919363081
influence of sprinklers, used to alleviate heat stress, on faecal shedding of e. coli o157:h7 and salmonella and antimicrobial susceptibility of salmonella and enterococcus in lactating dairy cattle.to determine the effect of sprinklers on faecal shedding of escherichia coli (e. coli) o157:h7 and salmonella in lactating dairy cattle.200919413804
effect of milk on antibacterial activity of tetracycline against escherichia coli and staphylococcus aureus isolated from bovine mastitis.the susceptibility of mastitis-causing escherichia coli and staphylococcus aureus to two commonly used antibiotics, tetracycline and penicillin g, was tested in raw milk and in muller-hinton (mh) broth by introducing a ph indicator, bromocresol purple, which was shown to be a simple, sensitive, and rapid method. the minimum inhibitory concentration (mic) of penicillin g in milk was the same as those in mh broth, whereas the mic of tetracycline in milk was 4 to 32 times that in mh. an irreversibl ...200919418049
inhibition of atpase activity of escherichia coli atp synthase by polyphenols.we have studied the inhibitory effect of five polyphenols namely, resveratrol, piceatannol, quercetin, quercetrin, and quercetin-3-beta-d glucoside on escherichia coli atp synthase. recently published x-ray crystal structures of bovine mitochondrial atp synthase inhibited by resveratrol, piceatannol, and quercetin, suggest that these compounds bind in a hydrophobic pocket between the gamma-subunit c-terminal tip and the hydrophobic inside of the surrounding annulus in a region critical for rotat ...200919375450
changes in escherichia coli o157:h7 numbers during holding on excised lean, fascia and fat beef surfaces at different temperatures.to investigate changes in escherichia coli o157:h7 numbers on excised beef carcass surfaces over 72 h at different temperatures.200919457044
occurrence of non-sorbitol fermenting, verocytotoxin-lacking escherichia coli o157 on cattle farms.escherichia coli o157 is often associated with hemorrhagic colitis and the hemolytic uremic syndrome (hus). the verocytotoxins are considered to be the major virulence determinants. however, vt-negative e. coli o157 were recently isolated from patients with hus. several transmission routes to humans are described, but cattle feces are the primary source from which both the food supply and the environment become contaminated with e. coli o157. in a prevalence study performed on dairy, beef, mixed ...200919299094
effect of intrauterine infusion of ceftiofur on uterine health and fertility in dairy cows.objectives were to determine the effects of intrauterine (i.u.) infusion of ceftiofur hydrochloride on uterine health and fertility of dairy cows already receiving pgf(2alpha) for estrous synchronization. holstein cows at 44 +/- 3 d in milk (dim) were blocked by parity and diagnosis of metritis in the first 14 dim and of other illnesses and, within each block, randomly assigned to 1 of 2 treatments: a single i.u. infusion of 125 mg of ceftiofur hydrochloride at 44 +/- 3 dim (ceftiofur, n = 396), ...200919307634
dynamics of endemic infectious diseases of animal and human importance on three dairy herds in the northeastern united states.endemic infectious diseases in dairy cattle are of significant concern to the industry as well as for public health because of their potential impact on animal and human health, milk and meat production, food safety, and economics. we sought to provide insight into the dynamics of important endemic infectious diseases in 3 northeastern us dairy herds. fecal samples from individual cows and various environmental samples from these farms were tested for the presence of major zoonotic pathogens (i. ...200919307664
successful application of the dual-vector system ii in creating a reliable phage-displayed combinatorial fab library.the dual-vector system-ii (dvs-ii), which allows efficient display of fab antibodies on phage, has been reported previously, but its practical applicability in a phage-displayed antibody library has not been verified. to resolve this issue, we created two small combinatorial human fab antibody libraries using the dvs-ii, and isolation of target-specific antibodies was attempted. biopanning of one antibody library, termed dvfab-1l library, which has a 1.3 x 10(7) combinatorial antibody complexity ...200919326078
escherichia coli o157:h7 strains isolated from environmental sources differ significantly in acetic acid resistance compared with human outbreak strains.a number of studies on the influence of acid on escherichia coli o157:h7 have shown considerable strain differences, but limited information has been reported to compare the acid resistance based on the different sources of e. coli o157:h7 isolates. the purpose of this study was to determine the survival of e. coli o157:h7 strains isolated from five sources (foods, bovine carcasses, bovine feces, water, and human) in 400 mm acetic acid solutions under conditions that are typical of acidified foo ...200919343937
escherichia coli o157:h7 lineages in healthy beef and dairy cattle and clinical human cases in alberta, canada.the aim of this study was to examine the prevalence and distribution of escherichia coli o157:h7 lineage-specific polymorphism assay (lspa) 6 genotypes from cattle (n = 313) and clinical human (n = 203) isolates from northern and southern alberta, canada, to understand possible associations of genotypes with host and geographic location. the majority of cattle isolates (feedlot and dairy) typed as lspa-6 111111 (72.2%), with proportionately higher lspa-6 222222 (19.4%) than other lspa-6 genotype ...200919343950
functions of manduca sexta hemolymph proteinases hp6 and hp8 in two innate immune pathways.serine proteinases in insect plasma have been implicated in two types of immune responses; that is, activation of prophenoloxidase (propo) and activation of cytokine-like proteins. we have identified more than 20 serine proteinases in hemolymph of the tobacco hornworm, manduca sexta, but functions are known for only a few of them. we report here functions of two additional m. sexta proteinases, hemolymph proteinases 6 and 8 (hp6 and hp8). hp6 and hp8 are each composed of an amino-terminal clip d ...200919487692
role of ceftiofur in selection and dissemination of blacmy-2-mediated cephalosporin resistance in salmonella enterica and commensal escherichia coli isolates from cattle.third-generation cephalosporin resistance of salmonella and commensal escherichia coli isolates from cattle in the united states is predominantly conferred by the cephamycinase cmy-2, which inactivates beta-lactam antimicrobial drugs used to treat a wide variety of infections, including pediatric salmonellosis. the emergence and dissemination of bla(cmy-2)(-)-bearing plasmids followed and may in part be the result of selection pressure imposed by the widespread utilization of ceftiofur, a third- ...200919376926
effect of tenderizers combined with organic acids on escherichia coli o157:h7 thermal resistance in non-intact beef.non-intact beef products include beef cuts that have been ground, mechanically tenderized, restructured, or have been injected with solutions to enhance tenderness and/or flavor. this study examined the effects of tenderizing salts and organic acids on thermal inactivation of escherichia coli o157:h7 in a ground beef model system simulating non-intact beef products. ground beef (95% lean; 700 g batches) was mixed (2 min) with nothing (c) or solutions (22 ml) of water (wa), calcium ascorbate (caa ...200919493583
validation of low-volume enrichment protocols for detection of escherichia coli o157 in raw ground beef components, using commercial kits.testing of beef destined for use in ground beef products for the presence of escherichia coli o157:h7 has become an important cornerstone of control and verification activities within many meat supply chains. validation of the ability of methods to detect low levels of e. coli o157:h7 is critical to confidence in test systems. many rapid methods have been validated against standard cultural methods for 25-g samples. in this study, a number of previously validated enrichment broths and commercial ...200919343962
recombinant chymosin used for exact and complete removal of a prochymosin derived fusion tag releasing intact native target protein.fusion tags add desirable properties to recombinant proteins, but they are not necessarily acceptable in the final products. ideally, fusion tags should be removed releasing the intact native protein with no trace of the tag. unique endoproteinases with the ability to cleave outside their own recognition sequence can potentially cleave at the boundary of any native protein. chymosin was recently shown to cleave a pro-chymosin derived fusion tag releasing native target proteins. in our hands, how ...200919388053
molecular analysis of escherichia coli from retail meats (2002-2004) from the united states national antimicrobial resistance monitoring system.the origins and virulence potential of meat product-associated escherichia coli are undefined.200919508167
therapeutic efficacy of mammary irrigation regimen in dairy cattle diagnosed with acute coliform mastitis.the objective of this field study was to determine the therapeutic efficacy of mammary irrigation for the treatment of dairy cattle diagnosed with acute coliform mastitis caused by gram-negative bacteria. additionally, the effects of different mammary irrigation regimen fluids such as ozone water and normal saline were compared. dairy cattle clinically diagnosed with acute coliform mastitis (n = 57) were enrolled in the study, randomly assigned to 1 of 3 groups, and received the following treatm ...200919346692
changing prevalence of o-serogroups and antimicrobial susceptibility among stec strains isolated from healthy dairy cows over a decade in japan between 1998 and 2007.the prevalence of stec in japan was examined using rectal stool samples taken from 932 healthy dairy cows from 123 farms in 11 prefectures between 2006 and 2007. screening with stx-pcrs revealed the prevalence to be 30.4% (283 animals), and stec strains were isolated from 111 animals. although ten o-serogroups (o8, o22, o84, o103, o111, o113, o116, o136, o153 and o157) were the major o-serogroup in healthy dairy cows in japan in 1998, half of the 118 selected stec strains were serotyped as o2, o ...200919346709
characteristics and virulence genes of escherichia coli isolated from septicemic calves in southeast of iran.virulence factors are associated with the capacity of e. coli strains to cause intestinal and extraintestinal infections. thirty one e. coli isolates were obtained from heart blood or internal organs of septicemic calves. the o serogroups of isolates were determined. pcr assays were performed to determine the phylogenetic groups and presence of specific virulence genes. fourteen (45.16%) isolates belonged to seven o serogroups (o8, o15, o20, o45, o78, o101 and o103) and 17 (54.83%) isolates were ...200919067217
chaperone action of a versatile small heat shock protein from methanococcoides burtonii, a cold adapted archaeon.the methanococcoides burtonii small heat shock protein (mb-shsp) is an alphab-crystallin homolog that delivers protein stabilizing and protective functions to model enzymes, presumably reflecting its role as a molecular chaperone in vivo. although the gene encoding mb-shsp was cloned from a cold-adapted microorganism, the mb-shsp is an efficient protein chaperone at temperatures far above the optimum growth temperature of m. burtonii. we show that mb-shsp can prevent aggregation in e. coli cell ...200918951410
stable expression of a gfp-bsd fusion protein in babesia bovis merozoites.transfection has been a valuable technique for elucidating gene function in many pathogens. while transient transfection of babesia spp. has been reported previously, stable integration of exogenous genes in babesia has proven difficult. in this study, a plasmid was designed to target integration of a gfp-bsd gene into the babesia bovis ef-1alpha locus. babesia bovis-infected erythrocytes of the biologically cloned mo7 strain were transfected by electroporation with either circular or linear pla ...200918831975
glycosphingolipids from bovine milk and milk fat globule membranes: a comparative study. adhesion to enterotoxigenic escherichia coli strains.several components of milk fat globule membranes (mfgms) have been reported to display beneficial health properties and some of them have been implicated in the defense of newborns against pathogens. these observations prompted us to determine the glycosphingolipid content of mfgms and their interaction with pathogens. a comparative study with whole milk components was also carried out. milk fat globules and mfgms were isolated from milk. gangliosides and neutral glycosphingolipids were obtained ...200918937626
evaluation of the control of pathogen load by an anti-salmonella bacterium in a herd of cattle with persistent salmonella infection.to determine whether an anti-salmonella bacterium is involved in control of pathogen load in persistently infected cattle herds.200919119953
monitoring major mastitis pathogens at the population level based on examination of bulk tank milk samples.the objective was to determine the proportions and bacterial counts of major mastitis pathogens in samples of bulk tank milk (btm), as well as to clarify the relationship between these bacteria counts and bulk tank somatic cell count (btscc). the purpose was to judge the importance of the counts of mammary pathogens for btscc at the population level. samples of btm were collected from 268 randomly selected anonymous dairy herds (with approximately 29 000 dairy cows). staphylococcus aureus, other ...200919121238
microbial aetiology of acute clinical mastitis and agent-specific risk factors.a nation wide study on the microbial aetiology of cases of acute clinical mastitis in swedish dairy cows was conducted with the aim to investigate changes in the microbial panorama compared to a previous study performed 1994-1995. another aim was to investigate some agent-specific environmental and individual risk factors. milk samples were collected from 987 udder quarter cases from 829 cows during six 2-month periods from may 2002 to april 2003, and data on risk factors and demography were col ...200919155148
intermittent escherichia coli o157:h7 colonisation at the terminal rectum mucosa of conventionally-reared lambs.in cattle, the lymphoid rich regions of the rectal-anal mucosa at the terminal rectum are the preferred site for escherichia coli o157:h7 colonisation. all cattle infected by rectal swab administration demonstrate long-term e. coli o157:h7 colonisation, whereas orally challenged cattle do not demonstrate long-term e. coli o157:h7 colonisation in all animals. oral, but not rectal challenge of sheep with e. coli o157:h7 has been reported, but an exact site for colonisation in sheep is unknown. to ...200918959839
synergistic role of curli and cellulose in cell adherence and biofilm formation of attaching and effacing escherichia coli and identification of fis as a negative regulator of curli.curli are adhesive fimbriae of escherichia coli and salmonella enterica. expression of curli (csga) and cellulose (bcsa) is co-activated by the transcriptional activator csgd. in this study, we investigated the contribution of curli and cellulose to the adhesive properties of enterohaemorragic (ehec) o157:h7 and enteropathogenic e. coli (epec) o127:h6. while single mutations in csga, csgd or bcsa in epec and ehec had no dramatic effect on cell adherence, double csgabcsa mutants were significantl ...200919187284
an investigation of the expression and adhesin function of h7 flagella in the interaction of escherichia coli o157 : h7 with bovine intestinal epithelium.enterohaemorrhagic escherichia coli o157 : h7 is a bacterial pathogen that can cause haemorrhagic colitis and haemolytic uremic syndrome. in the primary reservoir host, cattle, the terminal rectum is the principal site of e. coli o157 colonization. in this study, bovine terminal rectal primary epithelial cells were used to examine the role of h7 flagella in epithelial adherence. binding of a flic(h7) mutant o157 strain to rectal epithelium was significantly reduced as was binding of the flagella ...200919016776
factors influencing the presence and concentration of e. coli o157 and e. coli in farm waste on six cattle farms in north-west england.to investigate the factors influencing the presence and burden of escherichia coli o157 in farm wastes.200919200325
detection of bovine leukemia virus specific antibodies using recombinant p24-elisa.we developed an indirect elisa test (rp24-elisa) for the detection of antibodies against the p24 capsid protein of bovine leukemia virus (blv) in bovine serum samples. a recombinant p24 (rp24) was expressed in the escherichia coli expression system, purified in a nickel charged resin and used as antigen in the elisa test. a cut-off point was established considering the distribution of reactivity values obtained by rp24-elisa on a set of 555 field serum samples that were stated as double positive ...200919201553
escherichia coli o157:h7 survival, biofilm formation and acid tolerance under simulated slaughter plant moist and dry conditions.microorganisms persisting in slaughter plant environments may develop acid resistance and be translocated to other environmental surfaces or products. the objective of this study was to evaluate the potential of escherichia coli o157:h7 to form biofilms and maintain acid resistance, under different culture habituation scenarios, on stainless steel coupons (2 x 5 x 0.08 cm), in the presence of beef carcass decontamination runoff fluids (washings). coupons were stored in test tubes with unsteriliz ...200919028314
prevalence and impact of bacteriophages on the presence of escherichia coli o157:h7 in feedlot cattle and their environment.the relationship between endemic bacteriophages infecting e. coli o157:h7 (referred to as "phage") and levels of shedding of e. coli o157:h7 by cattle was investigated in two commercial feedlots in southern alberta, canada. between may and november 2007, 10 pens of cattle were monitored by collection of pooled fecal pats, water with sediment from troughs, manure slurry from the pen floor, and rectal fecal samples from individual animals (20 per pen) at two separate times. bacteriophages infectin ...200919139243
development of a real-time pcr assay with an internal amplification control for the screening of shiga toxin-producing escherichia coli in foods.to develop and evaluate a real-time pcr assay incorporating an internal amplification control (iac) suitable for the screening of shiga toxin (stx)-producing escherichia coli (stec) in foods.200919220740
association of escherichia coli j5-specific serum antibody responses with clinical mastitis outcome for j5 vaccinate and control dairy cattle.dairy cattle in two commercial holstein herds were randomly selected to be vaccinated twice with j5, at approximately 60 days and 28 days before the expected calving date, or to be untreated controls. based on whether milk production changed following clinical mastitis or whether cows were culled or died within 30 days after onset, 51 mastitis cases were classified as severe or mild. j5-specific antibody responses were evaluated by enzyme-linked immunosorbent assay of all 32 severe and 19 mild c ...200919052158
similar cefoxitin-resistance plasmids circulating in escherichia coli from human and animal sources.the aim of this study was to determine the molecular epidemiology of cefoxitin-resistance escherichia coli identified in cattle entering feedlots and determine if there were any similarities to e. coli causing human infections in canadian hospitals. a total of 51 e. coli were isolated from a total of 2483 cattle entering four feedlots in southern alberta, canada. dna fingerprinting using pulsed-field gel electrophoresis revealed thirty-two unique patterns with two major clusters observed compris ...200918824313
antimicrobial susceptibility of udder pathogens from cases of acute clinical mastitis in dairy cows.to investigate occurrence of acquired antimicrobial resistance in udder pathogens mics in staphylococcus aureus (n=211), coagulase-negative staphylococci (cns) (n=56), streptococcus uberis (n=113), streptococcus dysgalactiae (n=152), streptococcus agalactiae (n=6), escherichia coli (n=163), and klebsiella spp. (n=42) were determined using microdilution. isolates were from a nation wide survey employing strict inclusion criteria. presence of acquired resistance was evaluated by species-specific e ...200919058930
cdna, genomic sequence cloning and overexpression of ribosomal protein s25 gene (rps25) from the giant panda.rps25 is a component of the 40s small ribosomal subunit encoded by rps25 gene, which is specific to eukaryotes. studies in reference to rps25 gene from animals were handful. the giant panda (ailuropoda melanoleuca), known as a "living fossil", are increasingly concerned by the world community. studies on rps25 of the giant panda could provide scientific data for inquiring into the hereditary traits of the gene and formulating the protective strategy for the giant panda. the cdna of the rps25 clo ...200919101821
growth and stress responses of the earthworm eisenia fetida to escherichia coli o157:h7 in an artificial soil.escherichia coli o157:h7 is an intestine-inhabiting bacterium associated with many severe disease outbreaks worldwide. it may enter the soil environment with the excreta of infected animals (including horses, cattle and chickens) and human. earthworms are able to protect themselves against invading pathogens due to their efficient innate defense system. to better understand the mechanisms earthworms (eisenia fetida) utilize to defend and eliminate escherichia coli o157:h7, we examined the change ...200919232388
selection, characterization, and application of dna aptamers for the capture and detection of salmonella enterica serovars.sensitive and specific pre-analytical sample processing methods are needed to enhance our ability to detect and quantify food borne pathogens from complex food and environmental samples. in this study, dna aptamers were selected and evaluated for the capture and detection of salmonella enterica serovar. typhimurium. a total of 66 candidate sequences were enriched against s. typhimurium outer membrane proteins (omps) with counter-selection against escherichia coli omps and lipopolysaccharides (lp ...200919049862
screening of high-affinity scfvs from a ribosome displayed library using biacore biosensor.an experimental protocol was developed to screen high-affinity single-chain fv antibody fragments (scfvs) from a xanthomonas axonopodis pv. citri (xac) immunized ribosome display library using biacore biosensor. the screening methods involved immobilizing antigen [lipopolysaccharides (lps) of xac] on sensor chip hpa and then unpurified expression products of scfvs flowing over the immobilized sensor chip. the affinity-improved scfvs were selected based on dissociation rate constants (k (d)). thi ...200918574567
antimicrobial resistance of escherichia coli o26 and o111 isolates from cattle and their characteristics.the present study was to investigate antimicrobial resistance profiles of escherichia coli o26 and o111 from cattle and to characterize the virulence genes of the resistant isolates. this paper reports the high prevalence of antimicrobial resistant e. coli o26 and o111 from cattle. among 37 e. coli o26 and 25 e. coli o111 isolates from the fecal specimens obtained from cattle, 26 (70%) and 15 (60%) were resistant to at least one antibiotic, respectively. forty (98%) of the 41 resistant isolates ...200918992996
transmission and quantification of verocytotoxin-producing escherichia coli o157 in dairy cattle and calves.data from a field study of 14 months duration in a naturally colonized dairy herd and data from an experiment with calves were used to quantify transmission of verocytotoxin-producing escherichia coli (vtec o157) in cattle. for the latter, two groups of 10 calves were randomly assigned and put out in one of two pastures. from each group, five animals were experimentally inoculated with 109 c.f.u. o157 vtec and, considered infectious, put back in their group. each of the susceptible contact calve ...200918346284
[influence of gold and silver nanoparticles on atpase activity of native and rehydrated cells of escherichia coli].the expressed changes in the bacteria cells' plasma membrane atpase activity values cells of escherichia coli, which are used in the manufacturing of vaccines against cattle colienterotoxemia, have been revealed under the influence of lyophilization/rehydration process. influence of gold (average size 20, 30, 45 nm) and silver (30 nm) nanoparticles in different concentrations on atpase activity of native and rehydrated (delyophilized) bacteria cells has been studied. gold and silver nanoparticle ...200920387660
[quantitative risk model for verocytotoxigenic escherichia coli cross-contamination during homemade hamburger preparation].the objective of this study was to develop a quantitative risk model for verocytotoxigenic escherichia coil (vtec) cross-contamination during hamburger preparation at home. published scientific information about the disease was considered for the elaboration of the model, which included a number of routines performed during food preparation in kitchens. the associated probabilities of bacterial transference between food items and kitchen utensils which best described each stage of the process we ...200920085188
association of the amino acid motifs of bola-drb3 alleles with mastitis pathogens in japanese holstein cows.the association of the polymorphism of bovine leukocyte antigen (bola-drb3) genes, identified by the polymerase chain reaction sequence-based typing (pcr-sbt) method, with resistance and susceptibility to mastitis caused by streptococci, coagulase-negative staphylococci, escherichia coli and staphylococcus aureus was investigated. blood samples for dna extraction were collected from 170 holstein cows (129 mastitis and 41 healthy cows) from 5 districts in chiba prefecture, japan. susceptibility o ...200920163614
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