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association of campylobacter upsaliensis with persistent bloody diarrhea.campylobacter upsaliensis is a zoonotic, emerging pathogen that is not readily recovered in traditional stool culture. this case represents the first report of persistent bloody diarrhea with c. upsaliensis that was confirmed by filtration culture, pcr, and sequencing.201222915607
sodium metasilicate affects growth of campylobacter jejuni in fresh, boneless, uncooked chicken breast fillets stored at 4 degrees celsius for 7 days.the objectives of this study were to determine the antimicrobial effects of sodium metasilicate (sms) treatments against campylobacter jejuni in fresh, boneless, uncooked chicken breast fillets and to ascertain the effects of sms treatments on ph. the fillets were inoculated with c. jejuni, treated with 0% sms and no inoculum (negative control), 0% sms and inoculum (positive control), 1 and 2% sms solutions, and stored at 4 ± 1°c. all samples were analyzed after 0, 1, 3, 5, and 7 d storage for c ...201222912470
garlic derivative propyl propane thiosulfonate is effective against broiler enteropathogens in vivo.two experiments were carried out to study the effects of dietary supplementation with the garlic (allium sativum)-derived product propyl propane thiosulfonate (pts-o) on the intestinal log(10) number of copies of enteropathogens in broiler chickens, together with their intestinal morphology and growth performance. the additive had no significant effect on feed intake at any dose assayed. in experiment 1 (1 to 21 d of age), the bw of chickens fed on 45 mg of pts-o/kg of diet was higher (p < 0.01) ...201222912448
a bioinformatic strategy for the detection, classification and analysis of bacterial autotransporters.autotransporters are secreted proteins that are assembled into the outer membrane of bacterial cells. the passenger domains of autotransporters are crucial for bacterial pathogenesis, with some remaining attached to the bacterial surface while others are released by proteolysis. an enigma remains as to whether autotransporters should be considered a class of secretion system, or simply a class of substrate with peculiar requirements for their secretion. we sought to establish a sensitive search ...201222905239
differences in the fecal concentrations and genetic diversities of campylobacter jejuni populations among individual cows in two dairy herds.dairy cows have been identified as common carriers of campylobacter jejuni, which causes many of the human gastroenteritis cases reported worldwide. to design on-farm management practices that control the human infection sourced from dairy cows, the first step is to acquire an understanding of the excretion patterns of the cow reservoir. we monitored the same 35 cows from two dairy farms for c. jejuni excretion fortnightly for up to 12 months. the objective was to examine the concentration of c. ...201222904055
lacto-n-tetraose, fucosylation, and secretor status are highly variable in human milk oligosaccharides from women delivering preterm.breast milk is the ideal nutrition for term infants but must be supplemented to provide adequate growth for most premature infants. human milk oligosaccharides (hmos) are remarkably abundant and diverse in breast milk and yet provide no nutritive value to the infant. hmos appear to have at least two major functions: prebiotic activity (stimulation of the growth of commensal bacteria in the gut) and protection against pathogens. investigations of hmos in milk from women delivering preterm have be ...201222900748
guillain-barre syndrome after coronary artery bypass surgery.a case of the guillain-barré syndrome occurring after otherwise uneventful cardiac surgery using cardiopulmonary bypass is presented. though the guillain-barré syndrome has been reported after surgical procedures, there are very few case reports after cardiopulmonary bypass surgery in the literature. the exact pathophysiological cause of the syndrome is still unknown. however, the most widely accepted hypothesis is that the syndrome is the result of an immune-mediated process. cardiac surgery ma ...201222899666
campylobacter antimicrobial resistance in peru: a ten-year observational study.campylobacter jejuni and campylobacter coli are food-borne pathogens of great importance and feature prominently in the etiology of developing world enteritis and travellers' diarrhoea. increasing antimicrobial resistant campylobacter prevalence has been described globally, yet data from peru is limited. our objective was to describe the prevalence trends of fluoroquinolone and macrolide-resistant c. jejuni and c. coli stool isolates from three regions in peru over a ten-year period.201222898609
in silico directed mutagenesis identifies the cd81/claudin-1 hepatitis c virus receptor interface.hepatitis c virus (hcv) entry is dependent on host cell molecules tetraspanin cd81, scavenger receptor bi and tight junction proteins claudin-1 and occludin. we previously reported a role for cd81/claudin-1 receptor complexes in hcv entry; however, the molecular mechanism(s) driving association between the receptors is unknown. we explored the molecular interface between cd81 and claudin-1 using a combination of bioinformatic sequence-based modelling, site-directed mutagenesis and fluorescent re ...201222897233
yersinia pestis infection and laboratory conditions alter flea-associated bacterial communities.we collected oropsylla montana from rock squirrels, spermophilus varigatus, and infected a subset of collected fleas with yersinia pestis, the etiological agent of plague. we used bar-tagged dna pyrosequencing to characterize bacterial communities of wild, uninfected controls and infected fleas. bacterial communities within y. pestis-infected fleas were substantially more similar to one another than communities within wild or control fleas, suggesting that infection alters the bacterial communit ...201222895162
yersinia pestis infection and laboratory conditions alter flea-associated bacterial communities.we collected oropsylla montana from rock squirrels, spermophilus varigatus, and infected a subset of collected fleas with yersinia pestis, the etiological agent of plague. we used bar-tagged dna pyrosequencing to characterize bacterial communities of wild, uninfected controls and infected fleas. bacterial communities within y. pestis-infected fleas were substantially more similar to one another than communities within wild or control fleas, suggesting that infection alters the bacterial communit ...201222895162
inhibitory effects of 405 nm irradiation on chlamydia trachomatis growth and characterization of the ensuing inflammatory response in hela cells.chlamydia trachomatis is an intracellular bacterium that resides in the conjunctival and reproductive tract mucosae and is responsible for an array of acute and chronic diseases. a percentage of these infections persist even after use of antibiotics, suggesting the need for alternative treatments. previous studies have demonstrated anti-bacterial effects using different wavelengths of visible light at varying energy densities, though only against extracellular bacteria. we investigated the effec ...201222894815
application of orange essential oil as an antistaphylococcal agent in a dressing model.staphylococcus aureus is the pathogen most often and prevalently involved in skin and soft tissue infections. in recent decades outbreaks of methicillin-resistant s. aureus (mrsa) have created major problems for skin therapy, and burn and wound care units. topical antimicrobials are most important component of wound infection therapy. alternative therapies are being sought for treatment of mrsa and one area of interest is the use of essential oils. with the increasing interest in the use and app ...201222894560
[simple and rapid detection methods for campylobacters].campylobacter jejuni and campylobacter coli have been leading cause of food poisoning in terms of both number of patients and cases in japan. therefore these bacteria are recognized as one of the most important food-borne pathogens. since campylobacters are microaerobic, slow growing, biochemically unreactive, not only it takes some time but also there are several problems regarding isolation and identification of campylobacter spp. we found that cytolethal distending toxin (cdt) genes are ubiqu ...201222894063
[basic research for the control of campylobacter food poisoning].appropriate handling and controlled temperature prevent cross-contamination and proliferation of contaminants in foods, thereby reducing the incidences of food-borne gastroenteritis in japan. however, the incidence of campylobacter jejuni/coli infection did not markedly decrease and has become one of the major causes of food-borne diseases. c. jejuni and c. coli are widespread in warm-blooded domestic animals; therefore, food products may easily become contaminated during processing. c. jejuni a ...201222894062
salmonella "sops" up a preferred electron receptor in the inflamed intestine.the microbiota of the mammalian intestinal tract represents a formidable barrier to colonization by pathogens. to overcome this resistance to colonization, bacterial pathogens use virulence factors to induce intestinal inflammation, which liberates nutrients for selective use by the infecting microbe. studies of salmonella enterica serovar typhimurium (s. typhimurium) infection in a streptomycin-treated mouse colitis model show how virulence factor-induced inflammation can produce nutrients used ...201222893385
a picomolar transition state analogue inhibitor of mtan as a specific antibiotic for helicobacter pylori.campylobacter and helicobacter species express a 6-amino-6-deoxyfutalosine n-ribosylhydrolase (hpmtan) proposed to function in menaquinone synthesis. but-dadme-imma is a 36 pm transition state analogue of hpmtan, and the crystal structure of the enzyme-inhibitor complex reveals the mechanism of inhibition. but-dadme-imma has a mic(90) value of <8 ng/ml for helicobacter pylori growth but does not cause growth arrest in other common clinical pathogens, thus demonstrating potential as an h. pylori- ...201222891633
guillain-barré syndrome associated with pulmonary tuberculosis.a 47-year-old male smoker admitted complaining of progressive bilateral lower-limb and upper-limb weakness, without any numbness. he also complained of productive cough and fever for the last 2 weeks before admission. there were no nasal symptoms, sore throat, night sweats, loss of appetite and weight, nor did he have any gastrointestinal symptoms.201222891003
acid stress response and protein induction in campylobacter jejuni isolates with different acid tolerance.during the transmission route from poultry to the human host, the major foodborne pathogen c. jejuni may experience many types of stresses, including low ph caused by different acids. however, not all strains are equally sensitive to the stresses. the aim of this study was to investigate the response to acid stress of three sequenced c. jejuni strains with different acid tolerances using hcl and acetic acid.201222889088
the apo-structure of the low molecular weight protein-tyrosine phosphatase a (mptpa) from mycobacterium tuberculosis allows for better target-specific drug development.protein-tyrosine phosphatases (ptps) and protein-tyrosine kinases co-regulate cellular processes. in pathogenic bacteria, they are frequently exploited to act as key virulence factors for human diseases. mycobacterium tuberculosis, the causative organism of tuberculosis, secretes a low molecular weight ptp (lmw-ptp), mptpa, which is required for its survival upon infection of host macrophages. although there is otherwise no sequence similarity of lmw-ptps to other classes of ptps, the phosphate ...201222888002
influence of asellus aquaticus on escherichia coli, klebsiella pneumoniae, campylobacter jejuni and naturally occurring heterotrophic bacteria in drinking water.water lice, asellus aquaticus (isopoda), frequently occur in drinking water distribution systems where they are a nuisance to consumers and water utilities. whether they are solely an aesthetic problem or also affect the microbial water quality is a matter of interest. we studied the influence of a. aquaticus on microbial water quality in non-chlorinated drinking water in controlled laboratory experiments. pure cultures of the indicator organisms escherichia coli and klebsiella pneumoniae and th ...201222884244
farm and environmental distribution of campylobacter and salmonella in broiler flocks.the objective of this study was to determine the prevalence of campylobacter and salmonella in broilers and their distribution in the indoor and outdoor farm environment. nine hundred samples (400 faecal; 300 indoor environment; 200 outdoor environment), were collected from 10 individual broiler houses on 10 farms. campylobacter jejuni prevalence was significantly higher (p=0.003) in faeces (29.5%; 118/400) than the environment (0.8%; 4/500) in contrast to salmonella typhimurium from faecal (8.8 ...201322884004
anti-gq1b-negative miller fisher syndrome after campylobacter jejuni enteritis.miller fisher syndrome is a clinical variant of guillain-barré syndrome, characterized by acute-onset ophthalmoplegia, ataxia, and areflexia. it results from an immune response to a cross-reactive antigen between gq1b ganglioside in human neurons and lipo-oligosaccharides of certain bacteria, e.g., campylobacter jejuni. anti-gq1b antibody is a powerful diagnostic marker for miller fisher syndrome. however, only a small number of anti-gq1b-negative miller fisher syndrome cases are documented. a 1 ...201222883289
short-term and medium-term clinical outcomes of campylobacter concisus infection.there are only sparse data on the short-term and medium-term clinical impacts of campylobacter concisus infection. a clinical study was performed during a 2-year period to determine the clinical manifestations in c. concisus-positive adult patients. a case patient was defined as an adult patient (≥18 years) with a c. concisus-positive stool sample during the study period. clinical data were obtained with use of a questionnaire supplemented with the patients' medical records, if any. the short-te ...201222882347
a comparison of computational methods for identifying virulence factors.bacterial pathogens continue to threaten public health worldwide today. identification of bacterial virulence factors can help to find novel drug/vaccine targets against pathogenicity. it can also help to reveal the mechanisms of the related diseases at the molecular level. with the explosive growth in protein sequences generated in the postgenomic age, it is highly desired to develop computational methods for rapidly and effectively identifying virulence factors according to their sequence info ...201222880014
extreme water-related weather events and waterborne disease.global climate change is expected to affect the frequency, intensity and duration of extreme water-related weather events such as excessive precipitation, floods, and drought. we conducted a systematic review to examine waterborne outbreaks following such events and explored their distribution between the different types of extreme water-related weather events. four medical and meteorological databases (medline, embase, georef, pubmed) and a global electronic reporting system (promed) were searc ...201222877498
extreme water-related weather events and waterborne disease.global climate change is expected to affect the frequency, intensity and duration of extreme water-related weather events such as excessive precipitation, floods, and drought. we conducted a systematic review to examine waterborne outbreaks following such events and explored their distribution between the different types of extreme water-related weather events. four medical and meteorological databases (medline, embase, georef, pubmed) and a global electronic reporting system (promed) were searc ...201222877498
atlast4ss: a curated database for type iv secretion systems.the type iv secretion system (t4ss) can be classified as a large family of macromolecule transporter systems, divided into three recognized sub-families, according to the well-known functions. the major sub-family is the conjugation system, which allows transfer of genetic material, such as a nucleoprotein, via cell contact among bacteria. also, the conjugation system can transfer genetic material from bacteria to eukaryotic cells; such is the case with the t-dna transfer of agrobacterium tumefa ...201222876890
microbiological applications of high-resolution melting analysis.high-resolution melting (hrm) analysis uses real-time pcr instrumentation to interrogate dna sequence variation and is a low-cost, single-step, closed-tube method. here we describe hrm technology and provide examples of varied clinical microbiological applications to highlight the strengths and limitations of hrm analysis.201222875887
biosynthesis of the polymannose lipopolysaccharide o-antigens from escherichia coli serotypes o8 and o9a requires a unique combination of single- and multiple-active site mannosyltransferases.the escherichia coli o9a and o8 o-antigen serotypes represent model systems for the abc transporter-dependent synthesis of bacterial polysaccharides. the o9a and o8 antigens are linear mannose homopolymers containing conserved reducing termini (the primer-adaptor), a serotype-specific repeat unit domain, and a terminator. synthesis of these glycans occurs on the polyisoprenoid lipid-linked primer, undecaprenol pyrophosphoryl-glcpnac, by two conserved mannosyltransferases, wbdc and wbdb, and a se ...201222875852
epidemiological association of campylobacter jejuni groups with pathogenicity-associated genetic markers.campylobacter jejuni, the most leading cause for bacterial gastroenteritis worldwide, shows a high genetic diversity among its isolates. recently, we demonstrated the existence of six c. jejuni-groups by combining mlst with six genetic markers. these groups were further characterized by the detection of cj1321-cj1326, fucp, cj0178, cj0755/cfra, ceue, plda, cstii, and cstiii in order (i.) to show further associations between these different genetic markers and mlst ccs. moreover, different studie ...201222873291
avian resistance to campylobacter jejuni colonization is associated with an intestinal immunogene expression signature identified by mrna sequencing.campylobacter jejuni is the most common cause of human bacterial gastroenteritis and is associated with several post-infectious manifestations, including onset of the autoimmune neuropathy guillain-barré syndrome, causing significant morbidity and mortality. poorly-cooked chicken meat is the most frequent source of infection as c. jejuni colonizes the avian intestine in a commensal relationship. however, not all chickens are equally colonized and resistance seems to be genetically determined. we ...201222870198
identification of a novel genomic island conferring resistance to multiple aminoglycoside antibiotics in campylobacter coli.historically, the incidence of gentamicin resistance in campylobacter has been very low, but recent studies reported a high prevalence of gentamicin-resistant campylobacter isolated from food-producing animals in china. the reason for the high prevalence was unknown and was addressed in this study. pcr screening identified aminoglycoside resistance genes apha-3 and apha-7 and the aade-sat4-apha-3 cluster among 41 campylobacter isolates from broiler chickens. importantly, a novel genomic island c ...201222869568
transforming clinical microbiology with bacterial genome sequencing.whole-genome sequencing of bacteria has recently emerged as a cost-effective and convenient approach for addressing many microbiological questions. here, we review the current status of clinical microbiology and how it has already begun to be transformed by using next-generation sequencing. we focus on three essential tasks: identifying the species of an isolate, testing its properties, such as resistance to antibiotics and virulence, and monitoring the emergence and spread of bacterial pathogen ...201222868263
eukaryotic n-glycosylation occurs via the membrane-anchored c-terminal domain of the stt3p subunit of oligosaccharyltransferase.n-glycosylation is an essential and highly conserved protein modification. in eukaryotes, it is catalyzed by a multisubunit membrane-associated enzyme, oligosaccharyltransferase (ot). we report the high resolution structure of the c-terminal domain of eukaryotic stt3p. unlike its soluble β-sheet-rich prokaryotic counterparts, our model reveals that the c-terminal domain of yeast stt3p is highly helical and has an overall oblate spheroid-shaped structure containing a membrane-embedded region. anc ...201222865878
diversity in the protein n-glycosylation pathways within the campylobacter genus.the foodborne bacterial pathogen, campylobacter jejuni, possesses an n-linked protein glycosylation (pgl) pathway involved in adding conserved heptasaccharides to asparagine-containing motifs of >60 proteins, and releasing the same glycan into its periplasm as free oligosaccharides. in this study, comparative genomics of all 30 fully sequenced campylobacter taxa revealed conserved pgl gene clusters in all but one species. structural, phylogenetic and immunological studies showed that the n-glyco ...201222859570
luminal hypertonicity and acidity modulate colorectal afferents and induce persistent visceral hypersensitivity.carbohydrate malabsorption such as in lactose intolerance or enteric infection causes symptoms that include abdominal pain. because this digestive disorder increases intracolonic osmolarity and acidity by accumulation of undigested carbohydrates and fermented products, we tested whether these two factors (hypertonicity and acidity) would modulate colorectal afferents in association with colorectal nociception and hypersensitivity. in mouse colorectum-pelvic nerve preparations in vitro, afferent ...201222859365
prospective evaluation of a matrix-assisted laser desorption ionization-time of flight mass spectrometry system in a hospital clinical microbiology laboratory for identification of bacteria and yeasts: a bench-by-bench study for assessing the impact on time to identification and cost-effectiveness.matrix-assisted laser desorption ionization-time of flight mass spectrometry (maldi-tof ms) has been found to be an accurate, rapid, and inexpensive method for the identification of bacteria and yeasts. previous evaluations have compared the accuracy, time to identification, and costs of the maldi-tof ms method against standard identification systems or commercial panels. in this prospective study, we compared a protocol incorporating maldi-tof ms (maldi protocol) with the current standard ident ...201222855510
expression of sialic acids and other nonulosonic acids in leptospira.sialic acids are negatively charged nine carbon backbone sugars expressed on mammalian cell surfaces. sialic acids are part of a larger family of nonulosonic acid (nulo) molecules that includes pseudaminic and legionaminic acids. microbial expression of sialic acids and other nonulosonic acids has been shown to contribute to host-microbe interactions in a variety of contexts, including participation in colonization, immune subversion, and behaviors such as biofilm formation, autoagglutination an ...201222853805
multifunctional role of dextran sulfate sodium for in vivo modeling of intestinal diseases.inflammatory bowel diseases (ibds) are chronic, relapsing disorders that affect the gastrointestinal tract of millions of people and continue to increase in incidence each year. while several factors have been associated with development of ibds, the exact etiology is unknown. research using animal models of ibds is beginning to provide insights into how the different factors contribute to disease development. oral administration of dextran sulfate sodium (dss) to mice induces a reproducible exp ...201222853702
exploiting the nephrotoxic effects of venom from the sea anemone, phyllodiscus semoni, to create a hemolytic uremic syndrome model in the rat.in the natural world, there are many creatures with venoms that have interesting and varied activities. although the sea anemone, a member of the phylum coelenterata, has venom that it uses to capture and immobilise small fishes and shrimp and for protection from predators, most sea anemones are harmless to man. however, a few species are highly toxic; some have venoms containing neurotoxins, recently suggested as potential immune-modulators for therapeutic application in immune diseases. phyllo ...201222851928
modulation of toxin production by the flagellar regulon in clostridium difficile.we show in this study that toxin production in clostridium difficile is altered in cells which can no longer form flagellar filaments. the impact of inactivation of flic, cd0240, flif, flig, flim, and flhb-flir flagellar genes upon toxin levels in culture supernatants was assessed using cell-based cytotoxicity assay, proteomics, immunoassay, and immunoblotting approaches. each of these showed that toxin levels in supernatants were significantly increased in a flic mutant compared to that in the ...201222851750
sialoadhesin promotes rapid proinflammatory and type i ifn responses to a sialylated pathogen, campylobacter jejuni.sialoadhesin (sn) is a macrophage (mφ)-restricted receptor that recognizes sialylated ligands on host cells and pathogens. although sn is thought to be important in cellular interactions of mφs with cells of the immune system, the functional consequences of pathogen engagement by sn are unclear. as a model system, we have investigated the role of sn in mφ interactions with heat-killed campylobacter jejuni expressing a gd1a-like, sialylated glycan. compared to sn-expressing bone marrow-derived ma ...201222851711
zinc competition among the intestinal microbiota.bioavailable levels of trace metals, such as iron and zinc, for bacterial growth in nature are sufficiently low that most microbes have evolved high-affinity binding and transport systems. the microbe campylobacter jejuni lives in the gastrointestinal tract of chickens, the principal source of human infection. a high-affinity abc transporter for zinc uptake is required for campylobacter survival in chicken intestines in the presence of a normal microbiota but not when chickens are raised with a ...201222851657
guillain-barré syndrome and anti-ganglioside antibodies: a clinician-scientist's journey.guillain-barré syndrome (gbs) is the most frequent cause of acute flaccid paralysis. having seen my first gbs patient in 1989, i have since then dedicated my time in research towards understanding the pathogenesis of gbs. along with several colleagues, we identified igg autoantibodies against ganglioside gm1 in two patients with gbs subsequent to campylobacter jejuni enteritis. we proceeded to demonstrate molecular mimicry between gm1 and bacterial lipo-oligosaccharide of c. jejuni isolated from ...201222850724
a mutation in the pp2c phosphatase gene in a staphylococcus aureus usa300 clinical isolate with reduced susceptibility to vancomycin and daptomycin.methicillin-resistant staphylococcus aureus (mrsa) strains with reduced susceptibility to vancomycin (mic of 4 to 8 μg/ml) are referred to as vancomycin-intermediate s. aureus (visa). in this study, we characterized two isogenic usa300 s. aureus isolates collected sequentially from a single patient with endocarditis where the s. aureus isolate changed from being susceptible to vancomycin (vssa) (1 μg/ml) to visa (8 μg/ml). in addition, the visa isolate lost beta-lactamase activity and showed inc ...201222850507
effect of crust freezing applied alone and in combination with ultraviolet light on the survival of campylobacter on raw chicken.the application of crust freezing (cf) applied as a stand-alone treatment or in combination with ultraviolet (uv) light for reducing the level of artificially inoculated campylobacter jejuni on raw chicken was investigated. cf air temperatures of -5, -15 and -27 °c (±3 °c) with freezing times of 70, 15 and 6 min, respectively, were used. the level of c. jejuni on chicken was also examined following subsequent refrigerated (0-4 °c) storage at 3 and 7 days. all cf treatments resulted in significan ...201222850386
purification and characterization of a new bacteriocin active against campylobacter produced by lactobacillus salivarius smxd51.strain smxd51, isolated from chicken ceca and identified as lactobacillus salivarius, produced a component that inhibits the growth of gram-positive and gram-negative bacteria and especially campylobacter jejuni. the active peptide from the cell-free supernatant of lb. salivarius smxd51 was purified in three steps: (i) precipitation with 80% saturated ammonium sulfate, (ii) elution on a reversed phase spe upti-clean cartridge using different concentrations of acetonitrile, (iii) final purificati ...201222850384
tracing campylobacter jejuni strains along the poultry meat production chain from farm to retail by pulsed-field gel electrophoresis, and the antimicrobial resistance of isolates.in this study campylobacter jejuni isolates were recovered from birds, carcasses and carcass portions from two broiler chicken flocks and from equipment used for carcass and meat processing along the production chain from farms to retail stores. isolates were subjected to pulsed-field gel electrophoresis (pfge) using smai and kpni restriction enzymes and their antimicrobial susceptibilities were determined. c. jejuni was recovered from product and equipment used with both flocks at each point in ...201222850383
phylogenetic position of aquificales based on the whole genome sequences of six aquificales species.species belonging to the order aquificales are believed to be an early branching lineage within the bacteria. however, the branching order of this group in single-gene phylogenetic trees is highly variable; for example, it has also been proposed that the aquificales should be grouped with ε-proteobacteria. to investigate the phylogenetic position of aquificales at the whole-genome level, here we reconstructed the phylogenetic trees of 18 bacteria including six aquificales species based on the co ...201222844640
the complete genome sequence of bacteriophage cp21 reveals modular shuffling in campylobacter group ii phages.campylobacter group ii phages described so far share a high degree of sequence similarity. we report the 182,833-bp genomic sequence of the closely related group ii phage cp21 and show that it has a completely different genomic organization. as in other group ii phages, the cp21 genome is composed of large modules separated by long dna repeat regions which obviously trigger recombination and modular shuffling.201222843857
ischaemic colitis associated with oral contraceptive and bisacodyl use.a woman with a history of hip replacement presented 3 days prior to revision of the hip with abdominal pain, diarrhoea and haematochezia. these symptoms began 6 h after she began taking bisacodyl in preparation for her upcoming surgery. she was on low-dose estradiol for hormone replacement therapy (hrt). subsequent colonoscopy and biopsies were consistent with acute colonic ischaemia (ci). she was treated with intravenous fluids and antibiotics and discharged and told to stop hrt and bisacodyl. ...201222843752
inhibition of listeria monocytogenes atcc 19115 on ham steak by tea bioactive compounds incorporated into chitosan-coated plastic films.the consumer demands for better quality and safety of food products have given rise to the development and implementation of edible films. the use of antimicrobial films can be a promising tool for controlling l. monocytogenes on ready to eat products. the aim of this study was to develop effective antimicrobial films incorporating bioactive compounds from green and black teas into chitosan, for controlling l. monocytogenes atcc 19115 on vacuum-packaged ham steak. the effectiveness of these anti ...201222839307
real-time pcr for diagnosing helicobacter pylori infection in patients with upper gastrointestinal bleeding: comparison with other classical diagnostic methods.the aim of this study was to determine the diagnostic usefulness of quantification of the h. pylori genome in detection of infection in patients with upper gastrointestinal bleeding (ugb). a total of 158 consecutive patients with digestive disorders, 80 of whom had clinical presentation of ugb, were studied. the number of microorganisms was quantified using a real-time pcr system which amplifies the urease gene with an internal control for eliminating the false negatives. a biopsy sample from th ...201222837325
behaviour of non-stressed and stressed listeria monocytogenes and campylobacter jejuni cells on fresh chicken burger meat packaged under modified atmosphere and inoculated with protective culture.numerous investigations have provided evidence that chicken products are a source of listeria monocytogenes and campylobacter jejuni. different strategies applied in final products are needed to prevent consumers' contamination. in this work, the combination of modified atmosphere packaging (map) and protective culture to control the growth of freeze stressed and non-stressed l. monocytogenes and c. jejuni on fresh chicken meat burger was studied. meat burgers were inoculated with l. monocytogen ...201222831819
interaction effects between sender and receiver processes in indirect transmission of campylobacter jejuni between broilers.infectious diseases in plants, animals and humans are often transmitted indirectly between hosts (or between groups of hosts), i.e. via some route through the environment instead of via direct contacts between these hosts. here we study indirect transmission experimentally, using transmission of campylobacter jejuni (c. jejuni) between spatially separated broilers as a model system. we distinguish three stages in the process of indirect transmission; (1) an infectious "sender" excretes the agent ...201222831274
reactive oxygen species in the signaling and adaptation of multicellular microbial communities.one of the universal traits of microorganisms is their ability to form multicellular structures, the cells of which differentiate and communicate via various signaling molecules. reactive oxygen species (ros), and hydrogen peroxide in particular, have recently become well-established signaling molecules in higher eukaryotes, but still little is known about the regulatory functions of ros in microbial structures. here we summarize current knowledge on the possible roles of ros during the developm ...201222829965
rsfa (ybeb) proteins are conserved ribosomal silencing factors.the ybeb (duf143) family of uncharacterized proteins is encoded by almost all bacterial and eukaryotic genomes but not archaea. while they have been shown to be associated with ribosomes, their molecular function remains unclear. here we show that ybeb is a ribosomal silencing factor (rsfa) in the stationary growth phase and during the transition from rich to poor media. a knock-out of the rsfa gene shows two strong phenotypes: (i) the viability of the mutant cells are sharply impaired during st ...201222829778
multilocus sequence types of environmental campylobacter jejuni isolates and their similarities to those of human, poultry and bovine c. jejuni isolates.in this study, we investigated the multilocus sequence type (mlst) diversity and population genetics of campylobacter jejuni isolates collected from the natural waters (n = 57), wild birds (n = 37) and zoo animals (n = 19) in southern finland, the helsinki area and the helsinki zoo, respectively. on average, we found c. jejuni in 20%, 10.4% or 11.5% of the samples collected from natural waters, wild birds and zoo animals, respectively. high st diversity was detected in all three sources and 41.2 ...201322827634
viscosity dictates metabolic activity of vibrio ruber.little is known about metabolic activity of bacteria, when viscosity of their environment changes. in this work, bacterial metabolic activity in media with viscosity ranging from 0.8 to 29.4 mpas was studied. viscosities up to 2.4 mpas did not affect metabolic activity of vibrio ruber. on the other hand, at 29.4 mpas respiration rate and total dehydrogenase activity increased 8 and 4-fold, respectively. the activity of glucose-6-phosphate dehydrogenase (gpd) increased up to 13-fold at higher vis ...201222826705
oxyr activation in porphyromonas gingivalis in response to a hemin-limited environment.porphyromonas gingivalis is a gram-negative obligately anaerobic bacterium associated with several forms of periodontal disease, most closely with chronic periodontitis. previous studies demonstrated that oxyr plays an important role in the aerotolerance of p. gingivalis by upregulating the expression of oxidative-stress genes. increases in oxygen tension and in h(2)o(2) both induce activation of oxyr. it is also known that p. gingivalis requires hemin as an iron source for its growth. in this s ...201222825453
genomic and proteomic characterization of sumu, a mu-like bacteriophage infecting haemophilus parasuis.haemophilus parasuis, the causative agent of glässer's disease, is prevalent in swine herds and clinical signs associated with this disease are meningitis, polyserositis, polyarthritis, and bacterial pneumonia. six to eight week old pigs in segregated early weaning herds are particularly susceptible to the disease. insufficient colostral antibody at weaning or the mixing of pigs with heterologous virulent h. parasuis strains from other farm sources in the nursery or grower-finisher stage are con ...201222823751
mucosal reactive oxygen species decrease virulence by disrupting campylobacter jejuni phosphotyrosine signaling.reactive oxygen species (ros) play key roles in mucosal defense, yet how they are induced and the consequences for pathogens are unclear. we report that ros generated by epithelial nadph oxidases (nox1/duox2) during campylobacter jejuni infection impair bacterial capsule formation and virulence by altering bacterial signal transduction. upon c. jejuni invasion, ros released from the intestinal mucosa inhibit the bacterial phosphotyrosine network that is regulated by the outer-membrane tyrosine k ...201222817987
reduced spread of campylobacter jejuni in broiler chickens by stimulating the bird's natural barriers.we have tested the effect of feed structure and feeding regime to prevent the spread of the zoonotic pathogen campylobacter jejuni in broiler chicken flocks.201222817452
do globins in microaerophilic campylobacter jejuni confer nitrosative stress tolerance under oxygen limitation?the microaerophilic pathogen campylobacter jejuni possesses inducible systems for resisting no. two globins--cgb (a single-domain globin) and ctb (a truncated globin)--are up-regulated in response to no via the positively acting transcription factor nssr. our aims were to determine whether these oxygen-binding globins also function in severely oxygen-limited environments, as in the host. at growth-limiting oxygen transfer rates, bacteria were more s-nitrosoglutathione (gsno) sensitive, irrespect ...201322816769
planktonic aggregates of staphylococcus aureus protect against common antibiotics.bacterial cells are mostly studied during planktonic growth although in their natural habitats they are often found in communities such as biofilms with dramatically different physiological properties. we have examined another type of community namely cellular aggregates observed in strains of the human pathogen staphylococcus aureus. by laser-diffraction particle-size analysis (lda) we show, for strains forming visible aggregates, that the aggregation starts already in the early exponential gro ...201222815921
structural heterogeneity of terminal glycans in campylobacter jejuni lipooligosaccharides.lipooligosaccharides of the gastrointestinal pathogen campylobacter jejuni are regarded as a major virulence factor and are implicated in the production of cross-reactive antibodies against host gangliosides, which leads to the development of autoimmune neuropathies such as guillain-barré and fisher syndromes. c. jejuni strains are known to produce diverse los structures encoded by more than 19 types of los biosynthesis clusters. this study demonstrates that the final c. jejuni los structure can ...201222815868
diarrhea outbreak during u.s. military training in el salvador.infectious diarrhea remains a major risk to deployed military units worldwide in addition to their impact on travelers and populations living in the developing world. this report describes an outbreak of diarrheal illness in the u.s. military's 130(th) maneuver enhancement brigade deployed in san vicente, el salvador during a training and humanitarian assistance mission. an outbreak investigation team from u.s. naval medical research unit-six conducted an epidemiologic survey and environmental a ...201222815747
a longitudinal 6-year study of the molecular epidemiology of clinical campylobacter isolates in oxfordshire, united kingdom.temporal and seasonal trends in campylobacter genotypes causing human gastroenteritis were investigated in a 6-year study of 3,300 recent isolates from oxfordshire, united kingdom. genotypes (sequence types [st]) were defined using multilocus sequence typing and assigned to a clonal complex (a cluster of related strains that share four or more identical alleles with a previously defined central genotype). a previously undescribed clonal complex (st-464) was identified which, together with st-42, ...201222814466
campylobacter jejuni induces acute enterocolitis in gnotobiotic il-10-/- mice via toll-like-receptor-2 and -4 signaling.campylobacter jejuni is a leading cause of foodborne bacterial enterocolitis worldwide. investigation of immunopathology is hampered by a lack of suitable vertebrate models. we have recently shown that gnotobiotic mice as well as conventional il-10(-/-) animals are susceptible to c. jejuni infection and develop intestinal immune responses. however, clinical symptoms of c. jejuni infection were rather subtle and did not reflect acute bloody diarrhea seen in human campylobacteriosis.201222808254
glycerol monolaurate antibacterial activity in broth and biofilm cultures.glycerol monolaurate (gml) is an antimicrobial agent that has potent activity against gram-positive bacteria. this study examines gml antibacterial activity in comparison to lauric acid, in broth cultures compared to biofilm cultures, and against a wide range of gram-positive, gram-negative, and non-gram staining bacteria.201222808139
glycopp: a webserver for prediction of n- and o-glycosites in prokaryotic protein sequences.glycosylation is one of the most abundant post-translational modifications (ptms) required for various structure/function modulations of proteins in a living cell. although elucidated recently in prokaryotes, this type of ptm is present across all three domains of life. in prokaryotes, two types of protein glycan linkages are more widespread namely, n- linked, where a glycan moiety is attached to the amide group of asn, and o- linked, where a glycan moiety is attached to the hydroxyl group of se ...201222808107
recent advances in small bowel diseases: part i.as is the case in all parts of gastroenterology and hepatology, there have been many advances in our knowledge and understanding of small intestinal diseases. over 1000 publications were reviewed for 2008 and 2009, and the important advances in basic science as well as clinical applications were considered. in part i of this editorial review, seven topics are considered: intestinal development; proliferation and repair; intestinal permeability; microbiotica, infectious diarrhea and probiotics; d ...201222807604
restricted gene flow among hospital subpopulations of enterococcus faecium.enterococcus faecium has recently emerged as an important multiresistant nosocomial pathogen. defining population structure in this species is required to provide insight into the existence, distribution, and dynamics of specific multiresistant or pathogenic lineages in particular environments, like the hospital. here, we probe the population structure of e. faecium using bayesian-based population genetic modeling implemented in bayesian analysis of population structure (baps) software. the anal ...201222807567
distribution and genotypic characterization of campylobacter jejuni isolated from poultry in split and dalmatia county, croatia.consumption of poultry contaminated with campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. therefore, the aim of our study was to investigate the prevalence and genotype diversity of campylobacter jejuni in poultry meat intended for consumption in split and dalmatia county, which is the second biggest county in croatia. furthermore, we also wanted to discover possibly stable clones of c. jejuni appearing in different samples and periods of time, which ...201322805333
a ferritin mutant of mycobacterium tuberculosis is highly susceptible to killing by antibiotics and is unable to establish a chronic infection in mice.iron is an essential, elusive, and potentially toxic nutrient for most pathogens, including mycobacterium tuberculosis. due to the poor solubility of ferric iron under aerobic conditions, free iron is not found in the host. m. tuberculosis requires specialized iron acquisition systems to replicate and cause disease. it also depends on a strict control of iron metabolism and intracellular iron levels to prevent iron-mediated toxicity. under conditions of iron sufficiency, m. tuberculosis represse ...201222802345
evaluating the efficacy of an avian-specific probiotic to reduce the colonization of campylobacter jejuni in broiler chickens.campylobacteriosis is the most frequent zoonotic disease in humans worldwide, and the contaminated poultry meat by campylobacter jejuni can be considered one of the important sources of enteric infections in humans. the use of probiotics, which can help to improve the natural defense of animals against pathogenic bacteria, is an alternative and effective approach to antibiotic administration for livestock to reduce bacterial contamination. in vitro experiments showed that enterococcus faecium, p ...201222802174
benign ulcer of the right colon clinically misdiagnosed as carcinoma: an additional case.benign solitary ulcer of the colon is an uncommon lesion that was originally described by cruveilhier in 1832. its aetiology remains unknown, and there are no pathognomonic lesions or symptoms. diagnosis is made by exclusion; in fact, diseases such as specific infections (cytomegalovirus, campylobacter jejuni, entamoeba histolytica), common clinical conditions (acute appendicitis, diverticulitis, intestinal obstruction, inflammatory bowel disease), pharmacotherapy (non-steroidal anti-inflammator ...201222799054
campylobacter epidemiology: a descriptive study reviewing 1 million cases in england and wales between 1989 and 2011.to review campylobacter cases in england and wales over 2 decades and examine the main factors/mechanisms driving the changing epidemiology.201222798256
hypomorphic glycosyltransferase alleles and recoding at contingency loci influence glycan microheterogeneity in the protein glycosylation system of neisseria species.as more bacterial protein glycosylation systems are identified and characterized, a central question that arises is, what governs the prevalence of particular glycans associated with them? in addition, accumulating evidence shows that bacterial protein glycans can be subject to the phenomenon of microheterogeneity, in which variant glycan structures are found at specific attachment sites of a given glycoprotein. although factors underlying microheterogeneity in reconstituted expression systems h ...201222797763
nutritional immunity: transition metals at the pathogen-host interface.transition metals occupy an essential niche in biological systems. their electrostatic properties stabilize substrates or reaction intermediates in the active sites of enzymes, and their heightened reactivity is harnessed for catalysis. however, this heightened activity also renders transition metals toxic at high concentrations. bacteria, like all living organisms, must regulate their intracellular levels of these elements to satisfy their physiological needs while avoiding harm. it is therefor ...201222796883
bacillus anthracis inosine 5'-monophosphate dehydrogenase in action: the first bacterial series of structures of phosphate ion-, substrate-, and product-bound complexes.inosine 5'-monophosphate dehydrogenase (impdh) catalyzes the first unique step of the gmp branch of the purine nucleotide biosynthetic pathway. this enzyme is found in organisms of all three kingdoms. impdh inhibitors have broad clinical applications in cancer treatment, as antiviral drugs and as immunosuppressants, and have also displayed antibiotic activity. we have determined three crystal structures of bacillus anthracis impdh, in a phosphate ion-bound (termed "apo") form and in complex with ...201222788966
the campylobacter jejuni dps homologue is important for in vitro biofilm formation and cecal colonization of poultry and may serve as a protective antigen for vaccination.in this work, we investigated the campylobacter jejuni dps (dna binding protein from starved cells) gene for a role in biofilm formation and cecal colonization in poultry. in vitro biofilm formation assays were conducted with stationary-phase cells in cell culture plates under microaerophilic conditions. these studies demonstrated a significant (>50%) reduction in biofilm formation by the c. jejuni dps mutant compared to that by the wild-type strain. studies in poultry also demonstrated the impo ...201222787197
complete 6-deoxy-d-altro-heptose biosynthesis pathway from campylobacter jejuni: more complex than anticipated.the campylobacter jejuni capsule is important for colonization and virulence in various infection models. in most strains, the capsule includes a modified heptose whose biological role and biosynthetic pathway are unknown. to decipher the biosynthesis pathway for the 6-deoxy-d-altro-heptose of strain 81-176, we previously showed that the 4,6-dehydratase wcbk and the reductase wcag generated gdp-6-deoxy-d-manno-heptose, but the c3 epimerase necessary to form gdp-6-deoxy-d-altro-heptose was not id ...201222787156
epidemiologic overview of malignant lymphoma.malignant lymphoma encompasses a wide variety of distinct disease entities. it is generally more common in developed countries and less common in developing countries. the east asia region has one of the lowest incidence rates of malignant lymphoma. the incidence of malignant lymphoma around the world has been increasing at a rate of 3-4% over the last 4 decades, while some stabilization has been observed in developed countries in recent years. the reasons behind this lymphoma epidemic are poorl ...201222783355
gene expression analysis of toll-like receptor pathways in heterophils from genetic chicken lines that differ in their susceptibility to salmonella enteritidis.previously conducted studies using two chicken lines (a and b) show that line a birds have increased resistance to a number of bacterial and protozoan challenges and that heterophils isolated from line a birds are functionally more responsive. furthermore, when stimulated with toll-like receptor (tlr) agonists, heterophils from line a expressed a totally different cytokine and chemokine mrna expression pattern than heterophils from line b. a large-scale gene expression profile using an agilent 4 ...201222783275
cloning, expression, and antigenicity of 14 proteins from campylobacter jejuni.fourteen campylobacter jejuni genes--pora, cadf, omp18, dnak, flac, peb1, peb2, peb3, peb4, ahpc, groel, tuf, hipo, and cj0069--were cloned and expressed in escherichia coli bl21. the recombinant proteins were purified on histidine (his) and glutathione s-transferase (gst) trap columns using the äkta explorer 100 system. recombinant proteins were visualized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified using matrix-assisted laser desorption ionization time-of-fli ...201222779748
campylobacter jejuni translocation across intestinal epithelial cells is facilitated by ganglioside-like lipooligosaccharide structures.translocation across intestinal epithelial cells is an established pathogenic feature of the zoonotic bacterial species campylobacter jejuni. the number of c. jejuni virulence factors known to be involved in translocation is limited. in the present study, we investigated whether sialylation of c. jejuni lipooligosaccharide (los) structures, generating human nerve ganglioside mimics, is important for intestinal epithelial translocation. we here show that c. jejuni isolates expressing ganglioside- ...201222778098
role of rela and spot in burkholderia pseudomallei virulence and immunity.burkholderia pseudomallei is a gram-negative soil bacterium and the causative agent of melioidosis, a disease of humans and animals. it is also listed as a category b bioterrorism threat agent by the u.s. centers for disease control and prevention, and there is currently no melioidosis vaccine available. small modified nucleotides such as the hyperphosphorylated guanosine molecules ppgpp and pppgpp play an important role as signaling molecules in prokaryotes. they mediate a global stress respons ...201222778096
a convenient and robust in vivo reporter system to monitor gene expression in the human pathogen helicobacter pylori.thirty years of intensive research have significantly contributed to our understanding of helicobacter pylori biology and pathogenesis. however, the lack of convenient genetic tools, in particular the limited effectiveness of available reporter systems, has notably limited the toolbox for fundamental and applied studies. here, we report the construction of a bioluminescent h. pylori reporter system based on the photorhabdus luminescens luxcdabe cassette. the system is constituted of a promoterle ...201222773640
bacteremia caused by antimicrobial resistant campylobacter species at a medical center in taiwan, 1998-2008.this study was intended to delineate the clinical and microbiological characteristics of patients with bacteremia caused by campylobacter species.201222771419
real-time taqman polymerase chain reaction-based genus-identification and pyrosequencing-based species identification of campylobacter jejuni, c. coli, c. lari, c. upsaliensis, and c. fetus directly on stool samples.a new method was developed for campylobacter identification and applied directly on 599 stool samples from diarrhoeagenic patients. here, the gyrase b gene of campylobacter was targeted in a 2-step process: first, taqman polymerase chain reaction (pcr)-based identification of c. jejuni, c. coli, c. upsaliensis, c. lari, and c. fetus at the genus level, and, second, pyrosequencing-based identification at the species level. the taqman pcr method was compared to culturing and identified 87 campylob ...201222770773
estimating the relative roles of recombination and point mutation in the generation of single locus variants in campylobacter jejuni and campylobacter coli.single locus variants (slvs) are bacterial sequence types that differ at only one of the seven canonical multilocus sequence typing (mlst) loci. estimating the relative roles of recombination and point mutation in the generation of new alleles that lead to slvs is helpful in understanding how organisms evolve. the relative rates of recombination and mutation for campylobacter jejuni and campylobacter coli were estimated at seven different housekeeping loci from publically available mlst data. th ...201222767048
towards a pathogenic escherichia coli detection platform using multiplex sybr®green real-time pcr methods and high resolution melting analysis.escherichia coli is a group of bacteria which has raised a lot of safety concerns in recent years. five major intestinal pathogenic groups have been recognized amongst which the verocytotoxin or shiga-toxin (stx1 and/or stx2) producing e. coli (vtec or stec respectively) have received a lot of attention recently. indeed, due to the high number of outbreaks related to vtec strains, the european food safety authority (efsa) has requested the monitoring of the "top-five" serogroups (o26, o103, o111 ...201222761753
modification of the campylobacter jejuni n-linked glycan by eptc protein-mediated addition of phosphoethanolamine.campylobacter jejuni is the major worldwide cause of bacterial gastroenteritis. c. jejuni possesses an extensive repertoire of carbohydrate structures that decorate both protein and non-protein surface-exposed structures. an n-linked glycosylation system encoded by the pgl gene cluster mediates the synthesis of a rigidly conserved heptasaccharide that is attached to protein substrates or released as free oligosaccharide in the periplasm. removal of n-glycosylation results in reduced virulence an ...201222761430
in silico analysis of sequenced strains of clostridium difficile reveals a related set of conjugative transposons carrying a variety of accessory genes.the human gut pathogen clostridium difficile contains many conjugative transposons that have an array of accessory genes. in the current study, recently sequenced genomes were analyzed to identify new putative conjugative transposons. eleven new elements in 5 c. difficile strains were identified and all had a similar structure to the previously described elements ctn1, ctn5 and ctn7 in c. difficile strain 630. each element identified did however contain a new set of accessory genes compared with ...201222754747
noncanonical notch signaling modulates cytokine responses of dendritic cells to inflammatory stimuli.dendritic cell (dc)-derived cytokines play a key role in specifying adaptive immune responses tailored to the type of pathogen encountered and the local tissue environment. however, little is known about how dcs perceive the local environment. we investigated whether endogenous notch signaling could affect dc responses to pathogenic stimuli. we demonstrate that concurrent notch and tlr stimulation results in a unique cytokine profile in mouse bone-marrow derived dcs characterized by enhanced il- ...201222753939
carbohydrate binding and gene expression by in vitro and in vivo propagated campylobacter jejuni after immunomagnetic separation.campylobacter jejuni is an important human food-borne intestinal pathogen, however relatively little is known about its mechanisms of pathogenesis or pathogen-host interactions. to monitor changes in gene expression and glycan binding of c. jejuni within a common avian host, an immunomagnetic separation technique (ims) was utilised to directly isolate infecting c. jejuni 81116 from a chicken host. an average of 10(5) cells/g was re-isolated from chicken caecal samples by ims technique. the in vi ...201322753110
dnak dependence of the mycobacterial stress-responsive regulator hspr is mediated through its hydrophobic c-terminal tail.hspr is a repressor known to control expression of heat shock operons in a number of eubacteria. in mycobacteria and in several other actinobacteria, this protein is synthesized from the dnakje-hspr operon. previous investigations revealed that hspr binds to the operon promoter, thereby controlling its expression in an autoregulatory manner. dnak, which is a product of the same operon, further aids this autoregulatory process by stimulating the operator binding activity of hspr. the molecular me ...201222753065
antimicrobial activities of fidaxomicin.fidaxomicin is bactericidal against clostridium difficile. the combined results of 8 in vitro studies of 1323 c. difficile isolates showed the minimum inhibitory concentration (mic) range of fidaxomicin to be ≤ 0.001-1 μg/ml, with a maximum mic for inhibition of 90% of organisms (mic(90)) of 0.5 μg/ml. isolates from 2 phase iii clinical trials demonstrated that fidaxomicin mics of baseline isolates did not predict clinical cure, failure, or recurrence of c. difficile infections. no resistance to ...201222752863
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