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fluorescence-based comparative evaluation of bactericidal potency and food application potential of anti-listerial bacteriocin produced by lactic acid bacteria isolated from indigenous samples.the aim of the present study was to ascertain the potency of anti-listerial bacteriocin produced by lactic acid bacteria (lab) isolated from indigenous samples of dahi, dried fish, and salt-fermented cucumber. a total of 231 lab isolates were obtained from the samples, of which 51 isolates displayed anti-listerial activity. the anti-listerial lab were identified by pcr as lactobacillus sp., pediococcus sp., enterococcus sp., and lactococcus sp. pcr also enabled the detection of class iia bacteri ...201226781854
comparison of the potency of the lipid ii targeting antimicrobials nisin, lacticin 3147 and vancomycin against gram-positive bacteria.while nisin (lantibiotic), lacticin 3147 (lantibiotic) and vancomycin (glycopeptides) are among the best studied lipid ii-binding antimicrobials, their relative activities have never been compared. nisin and lacticin 3147 have been employed/investigated primarily as food preservatives, although they do have potential in terms of veterinary and clinical applications. vancomycin is used exclusively in clinical therapy. we reveal a higher potency for lacticin 3147 (mic 0.95-3.8 μg/ml) and vancomyci ...201226781852
novel polymer anchored cr(iii) schiff base complexes: synthesis, characterization and antimicrobial properties.new polymer-bound schiff bases and cr(iii) complexes have been synthesized by the reaction of 4-benzyloxybenzaldehyde, polymer-bound with 2-aminophenol, 2-amino-4-chlorophenol and 2-amino-4-methylphenol. the structure of polymeric-schiff bases and their cr(iii) complexes have been characterized by elemental analyses, magnetic measurements, ir, uv-vis, tg-dta and (1)h-nmr. all these compounds have also been investigated for antibacterial activity by the well-diffusion method against staphylococcu ...201222622060
development of sandwich enzyme-linked immunosorbent assay for the detection of cronobacter muytjensii (formerly called enterobacter sakazakii).this study aimed to produce a polyclonal antibody against cronobacter muytjensii (c. muytjensii, formerly called enterobacter sakazakii) and to develop an immunoassay for its detection. the optimum production of rabbit anti-c. muytjensii immunoglobulin g (igg) and chicken anti-c. muytjensii igy was reached in weeks 8 and 9, respectively. purification of rabbit anti-c. muytjensii igg from immunized rabbit sera was accomplished using the caprylic acid and ammonium sulfate precipitation method. as ...201222519814
microbiological performance of a food safety management system in a food service operation.the microbiological performance of a food safety management system in a food service operation was measured using a microbiological assessment scheme as a vertical sampling plan throughout the production process, from raw materials to final product. the assessment scheme can give insight into the microbiological contamination and the variability of a production process and pinpoint bottlenecks in the food safety management system. three production processes were evaluated: a high-risk sandwich p ...201222488059
characterization of lysb4, an endolysin from the bacillus cereus-infecting bacteriophage b4.bacillus cereus is a foodborne pathogen that causes emetic or diarrheal types of food poisoning. the incidence of b. cereus food poisoning has been gradually increasing over the past few years, therefore, biocontrol agents effective against b. cereus need to be developed. endolysins are phage-encoded bacterial peptidoglycan hydrolases and have received considerable attention as promising antibacterial agents.201222416675
volatile organic compounds from centaurium erythraea rafn (croatia) and the antimicrobial potential of its essential oil.gc and ms were used for the analysis of croatian centaurium erythraea rafn essential oil (obtained by hydrodistillation) and headspace (applying headspace solid-phase microextraction). the headspace contained numerous monoterpene hydrocarbons (the major ones were terpinene-4-ol, methone, p-cymene, γ-terpinene and limonene). oxygenated monoterpenes were present in the headspace and oil, while 1,8-cineole, bornyl acetate and verbenone were present only in the headspace. high headspace percentages ...201222349896
assessment of microbial quality of commercial and home-made tiger-nut beverages.we aimed to assess the microbiological quality of spanish commercial tiger-nut beverages as well as home-made samples collected from supermarkets, street vendors, juice bars and ice-cream parlours located in valencia.201222283438
evaluation of food protective property of five natural products using fresh-cut apple slice model.the present study evaluated the antioxidant (aa), antimicrobial and preservation effects of five plant derived natural products viz., rosmarinic acid (ra), p-coumaric acid (pca), trans-cinnamic acid (tca), hydroxyphenyllactic acid (hpa) and caffeic acid (ca) along with synthetic compounds (ascorbic acid, gallic acid, citric acid and bha) on fresh cut apple slices. antimicrobial efficacy of these compounds against bacillus licheniformis, pseudomonas vulgaris, shigella boydii, salmonella typhi, st ...201222530437
mannose receptor, c type 1 contributes to bacterial uptake by placental trophoblast giant cells.during pregnancy, maternal immune function is strictly controlled and immune tolerance is induced. trophoblast giant (tg) cells exhibit phagocytic activity and show macrophage-like activity against microorganisms in the placenta. however, details of molecular receptors and mechanisms for uptake by tg cells have not been clarified. in this study, we investigated the involvement of the mannose receptor, c type 1 (mrc1), in the uptake of the abortion-inducible bacterium listeria monocytogenes and a ...201223163874
cytotoxicity of bacterial metabolic products, including listeriolysin o, on leukocyte targets.bacterial toxins can exhibit anticancer activities. here we investigated the anticancer effects of the listeriolysin o toxin produced by listeria monocytogenes. we found that supernatants of listeria monocytogenes strains (wild type, 1189, and 1190) were cytotoxic to the jurkat cell line and human peripheral blood mononuclear cells (pbmc) in a concentration-dependent manner. the supernatant of strain 1044, not producing listeriolysin o, was inactive. the supernatants of listeria strains were als ...201223091365
isolation of a bacteriocin-producing lactococcus lactis and application of its bacteriocin to manage spoilage bacteria in high-value marine fish under different storage temperatures.the bacteriocins of lactic acid bacteria have considerable potential for biopreservation. the lactococcus lactis strain psy2 (genbank account no. jf703669) isolated from the surface of marine perch perca flavescens produced antibacterial activity against pathogenic and spoilage-causing gram-positive and gram-negative bacteria viz. arthrobacter sp., acinetobacter sp., bacillus subtilis, escherichia coli, listeria monocytogenes, pseudomonas aeruginosa and staphylococcus aureus and possessed broad ...201222555500
expression of virulence genes by listeria monocytogenes j0161 in natural environment.majority of studies concerning the gene expression of listeria monocytogenes have been done on pure culture states. our objective was to study l.monocytogenes in a co-cultured state and to understand if microbes in their natural state of existence are different in their expression than that of the purely cultured lab grown forms. for a long period discussions have been on the expression of prfa, (which is a virulence gene regulator) in a mammalian host and its role in causing the switch from a s ...201224031897
ifn-β expression is directly activated in human neutrophils transfected with plasmid dna and is further increased via tlr-4-mediated signaling.upon lps binding, tlr4 activates a myd88-dependent pathway leading to the transcriptional activation of proinflammatory genes, as well as a myd88-independent/trif-dependent pathway, responsible for the transcriptional induction of ifn-β. previous findings delineated that human neutrophils are unable to induce the transcription of ifn-β in response to tlr4 stimulation. because neutrophils do not express protein kinase c ε, a molecule recently reported as essential for initiating the myd88-indepen ...201222730532
invasive escherichia coli vaccines expressing brucella melitensis outer membrane proteins 31 or 16 or periplasmic protein bp26 confer protection in mice challenged with b. melitensis.because of the serious economic and medical consequences of brucellosis, efforts are to prevent infection of domestic animals through vaccines. many disadvantages are associated with the current brucella melitensis rev.1 vaccine prompting development of alternative vaccines and delivery. escherichia coli (dh5α) was engineered to express a plasmid containing the inv gene from yersinia pseudotuberculosis and the hly gene from listeria monocytogenes. these recombinant invasive e. coli expressing b. ...201222546330
inhibition of bacterial biofilm formation and swarming motility by a small synthetic cationic peptide.biofilms cause up to 80% of infections and are difficult to treat due to their substantial multidrug resistance compared to their planktonic counterparts. based on the observation that human peptide ll-37 is able to block biofilm formation at concentrations below its mic, we screened for small peptides with antibiofilm activity and identified novel synthetic cationic peptide 1037 of only 9 amino acids in length. peptide 1037 had very weak antimicrobial activity, but at 1/30th the mic the peptide ...201222354291
differential expression of virulence and stress fitness genes during interaction between listeria monocytogenes and bifidobacterium longum.bifidobacterium is well known to have an inhibitory effect on the survival, growth, and proliferation of various foodborne pathogens, but the mechanism of the molecular action of b. longum in blocking the invasion of listeria monocytogenes is not yet well defined. in the present study, following rna extraction and cdna synthesis, differential expression of virulence and stress fitness genes in l. monocytogenes and b. longum was determined by real-time pcr. the results indicate that l. monocytoge ...201222484935
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
campylobacter, salmonella, listeria monocytogenes, verotoxigenic escherichia coli, and escherichia coli prevalence, enumeration, and subtypes on retail chicken breasts with and without skin.this study examined the prevalence, counts, and subtypes of campylobacter, salmonella, listeria monocytogenes, verotoxigenic escherichia coli (vtec), and e. coli on raw retail chicken breast with the skin on versus the skin off. from january to december 2007, 187 raw skin-on chicken breasts and 131 skin-off chicken breasts were collected from randomly selected retail grocery stores in the region of waterloo, ontario, canada. campylobacter isolates were recovered from a higher proportion of the s ...201222221353
antimicrobial screening of some egyptian plants and active flavones from lagerstroemia indica leaves.one hundred and twenty four plant extracts were evaluated for their antimicrobial activity against four pathogenic bacteria (staphylococcus aureus (atcc 8095), salmonella enteritides (atcc 13076), escherichia coli (atcc 25922), and listeria monocytogenes (atcc 15313)) and candida albicans yeast (atcc 10231) using the disk diffusion and broth microdilution methods. of the plant extracts, fourteen exhibited antimicrobial activity against two or more of the five microorganisms tested. only the meth ...201223006992
production of human lactoferrin in animal milk.genetic constructs containing the human lactoferrin (hlf) gene were created within a joint program of russian and belorussian scientists. using these constructs, transgenic mice were bred (the maximum hlf concentration in their milk was 160 g/l), and transgenic goats were also generated (up to 10 g/l hlf in their milk). experimental goatherds that produced hlf in their milk were also bred, and the recombinant hlf was found to be identical to the natural protein in its physical and chemical prope ...201222360490
lessons from anaplasma phagocytophilum: chromatin remodeling by bacterial effectors.bacterial pathogens can alter global host gene expression via histone modifications and chromatin remodeling in order to subvert host responses, including those involved with innate immunity, allowing for bacterial survival. shigella flexneri, listeria monocytogenes, chlamydia trachomatis, and anaplasma phagocytophilum express effector proteins that modify host histones and chromatin structure. a. phagocytophilum modulates granulocyte respiratory burst in part by dampening transcription of sever ...201223082961
phylogenetic analysis of antimicrobial lactic acid bacteria from farmed seabass dicentrarchus labrax.the use of lactic acid bacteria (lab) in the prevention or reduction of fish diseases is receiving increasing attention. in the present study, 47 lab strains were isolated from farmed seabass ( dicentrarchus labrax ) and were phenotypically and phylogenetically analysed by 16s rdna and randomly amplified polymorphic dna - polymerase chain reaction (rapd-pcr). their antimicrobial effect was tested in vitro against a wide variety of pathogenic and spoilage bacteria. most of the strains isolated we ...201222439634
no effect of wolbachia on resistance to intracellular infection by pathogenic bacteria in drosophila melanogaster.multiple studies have shown that infection with the endosymbiotic bacterium wolbachia pipientis confers drosophila melanogaster and other insects with resistance to infection by rna viruses. studies investigating whether wolbachia infection induces the immune system or confers protection against secondary bacterial infection have not shown any effect. these studies, however, have emphasized resistance against extracellular pathogens. since wolbachia lives inside the host cell, we hypothesized th ...201222808174
age-associated alterations in cd8α+ dendritic cells impair cd8 t-cell expansion in response to an intracellular bacterium.age-associated decline in immunity to infection has been documented across multiple pathogens, yet the relative contributions of the aged priming environment and of lymphocyte-intrinsic defects remain unclear. to address the impact of the aging environment on t-cell priming, adult naïve ot-i tcr transgenic cd8 t cells, specific for the h-2kb-restricted immunodominant ova(257-264) epitope, were transferred into adult or old recipient mice infected with the recombinant intracellular bacterium list ...201222862959
single chain mhc i trimer-based dna vaccines for protection against listeria monocytogenes infection.to circumvent limitations of poor antigen presentation and immunogenicity of dna vaccines that target induction of cd8(+) t cell immunity, we have generated single chain mhc i trimers (mhc i scts) composed of a single polypeptide chain with a linear composition of antigenic peptide, β2-microglobulin, and heavy chain of a mhc class i molecule connected by flexible linkers. because of its pre-assembled nature, the sct presents enhanced expression and presentation of the antigenic peptide/mhc compl ...201222285270
intracerebral hemorrhages in adults with community associated bacterial meningitis in adults: should we reconsider anticoagulant therapy?to study the incidence, clinical presentation and outcome of intracranial hemorrhagic complications in adult patients with community associated bacterial meningitis.201223028898
a new hybrid bacteriocin, ent35-mccv, displays antimicrobial activity against pathogenic gram-positive and gram-negative bacteria.bacteriocins and microcins are ribosomally synthesized antimicrobial peptides that are usually active against phylogenetically related bacteria. thus, bacteriocins are active against gram-positive while microcins are active against gram-negative bacteria. the narrow spectrum of action generally displayed by bacteriocins from lactic acid bacteria represents an important limitation for the application of these peptides as clinical drugs or as food biopreservatives. the present study describes the ...201223650575
listeria monocytogenes triggers the cell surface expression of gp96 protein and interacts with its n terminus to support cellular infection.listeria monocytogenes is an intracellular food-borne pathogen causing listeriosis in humans. this bacterium deploys an arsenal of virulence factors that act in concert to promote cellular infection. bacterial surface proteins are of primary importance in the process of host cell invasion. they interact with host cellular receptors, inducing/modulating specific cellular responses. we previously identified vip, a listeria surface protein covalently attached to the bacterial cell wall acting as a ...201223109341
detection and identification of salmonella enterica, escherichia coli, and shigella spp. via pcr-electrospray ionization mass spectrometry: isolate testing and analysis of food samples.an assay to identify the common food-borne pathogens salmonella, escherichia coli, shigella, and listeria monocytogenes was developed in collaboration with ibis biosciences (a division of abbott molecular) for the plex-id biosensor system, a platform that uses electrospray ionization mass spectroscopy (esi-ms) to detect the base composition of short pcr amplicons. the new food-borne pathogen (fbp) plate has been experimentally designed using four gene segments for a total of eight amplicon targe ...201223001674
prevalence and relative risk of cronobacter spp., salmonella spp., and listeria monocytogenes associated with the body surfaces and guts of individual filth flies.although flies are important vectors of food-borne pathogens, there is little information to accurately assess the food-related health risk of the presence of individual flies, especially in urban areas. this study quantifies the prevalence and the relative risk of food-borne pathogens associated with the body surfaces and guts of individual wild flies. one hundred flies were collected from the dumpsters of 10 randomly selected urban restaurants. flies were identified using taxonomic keys before ...201222941079
highly specific fiber optic immunosensor coupled with immunomagnetic separation for detection of low levels of listeria monocytogenes and l. ivanovii.immunomagnetic separation (ims) and immunoassays are widely used for pathogen detection. however, novel technology platforms with highly selective antibodies are essential to improve detection sensitivity, specificity and performance. in this study, monoclonal antibodies (mabs) against internalin a (inla) and p30 were generated and used on paramagnetic beads of varying diameters for concentration, as well as on fiber-optic sensor for detection.201223176167
expanding the use of a fluorogenic method to determine activity and mode of action of bacillus thuringiensis bacteriocins against gram-positive and gram-negative bacteria.previously we described a rapid fluorogenic method to measure the activity of five bacteriocins produced by mexican strains of bacillus thuringiensis against b. cereus 183. here we standardize this method to efficiently determine the activity of bacteriocins against both gram-positive and gram-negative bacteria. it was determined that the crucial parameter required to obtain reproducible results was the number of cells used in the assay, that is, ~4 × 10(8) cell/ml and ~7 × 10(8) cell/ml, respec ...201222919330
functional characterization of the infection-inducible peptide edin in drosophila melanogaster.drosophila is a well-established model organism for studying innate immunity because of its high resistance against microbial infections and lack of adaptive immunity. in addition, the immune signaling cascades found in drosophila are evolutionarily conserved. upon infection, activation of the immune signaling pathways, toll and imd, leads to the expression of multiple immune response genes, such as the antimicrobial peptides (amps). previously, we identified an uncharacterized gene edin among t ...201222606343
microrna response to listeria monocytogenes infection in epithelial cells.micrornas represent a family of very small non-coding rnas that control several physiologic and pathologic processes, including host immune response and cancer by antagonizing a number of target mrnas. there is limited knowledge about cell expression and the regulatory role of micrornas following bacterial infections. we investigated whether infection with a gram-positive bacterium leads to altered expression of micrornas involved in the host cell response in epithelial cells. caco-2 cells were ...201222312311
sumoylation at the host-pathogen interface.many viral proteins have been shown to be sumoylated with corresponding regulatory effects on their protein function, indicating that this host cell modification process is widely exploited by viral pathogens to control viral activity. in addition to using sumoylation to regulate their own proteins, several viral pathogens have been shown to modulate overall host sumoylation levels. given the large number of cellular targets for sumo addition and the breadth of critical cellular processes that a ...201223795346
allosteric mutants show that prfa activation is dispensable for vacuole escape but required for efficient spread and listeria survival in vivo.the transcriptional regulator prfa controls key virulence determinants of the facultative intracellular pathogen listeria monocytogenes. prfa-dependent gene expression is strongly induced within host cells. while the basis of this activation is unknown, the structural homology of prfa with the camp receptor protein (crp) and the finding of constitutively activated prfa* mutants suggests it may involve ligand-induced allostery. here, we report the identification of a solvent-accessible cavity wit ...201222646689
fatty acids regulate stress resistance and virulence factor production for listeria monocytogenes.fatty acids (fas) are the major structural component of cellular membranes, which provide a physical and chemical barrier that insulates intracellular reactions from environmental fluctuations. the native composition of membrane fas establishes the topological and chemical parameters for membrane-associated functions and is therefore modulated diligently by microorganisms especially in response to environmental stresses. however, the consequences of altered fa composition during host-pathogen in ...201222843841
membrane damage during listeria monocytogenes infection triggers a caspase-7 dependent cytoprotective response.the cysteine protease caspase-7 has an established role in the execution of apoptotic cell death, but recent findings also suggest involvement of caspase-7 during the host response to microbial infection. caspase-7 can be cleaved by the inflammatory caspase, caspase-1, and has been implicated in processing and activation of microbial virulence factors. thus, caspase-7 function during microbial infection may be complex, and its role in infection and immunity has yet to be fully elucidated. here w ...201222807671
optimizing the balance between host and environmental survival skills: lessons learned from listeria monocytogenes.environmental pathogens - organisms that survive in the outside environment but maintain the capacity to cause disease in mammals - navigate the challenges of life in habitats that range from water and soil to the cytosol of host cells. the bacterium listeria monocytogenes has served for decades as a model organism for studies of host-pathogen interactions and for fundamental paradigms of cell biology. this ubiquitous saprophyte has recently become a model for understanding how an environmental ...201222827306
phosphatidylinositol 5-phosphatase oculocerebrorenal syndrome of lowe protein (ocrl) controls actin dynamics during early steps of listeria monocytogenes infection.listeria monocytogenes is a bacterial pathogen that induces its own entry into a broad range of mammalian cells through interaction of the bacterial surface protein inlb with the cellular receptor met, promoting an actin polymerization/depolymerization process that leads to pathogen engulfment. phosphatidylinositol bisphosphate (pi[4,5]p(2)) and trisphosphate (pi[3,4,5]p(3)) are two major phosphoinositide species that function as molecular scaffolds, recruiting cellular effectors that regulate a ...201222351770
[type i interferon and bacterial infection].interferons (ifns) are cytokines playing an important role in immune responses. interferons are classified into two distinct types according to specific interferon receptors(ifnr). type i ifns include ifn-α and ifn-β, whereas ifn-γ is type ii ifn. it is well known that type i ifns have important roles in the host defense against viruses through activation of interferon receptor a (ifnar). however, many recent studies have also demonstrated that type i ifns have effects on immune responses to bac ...201222927085
hiv-1 integrase inhibitor-inspired antibacterials targeting isoprenoid biosynthesis.we report the discovery of antibacterial leads, keto- and diketo-acids, targeting two prenyl transferases: undecaprenyl diphosphate synthase (upps) and dehydrosqualene synthase (crtm). the leads were suggested by the observation that keto- and diketo-acids bind to the active site mg(2+)/asp domain in hiv-1 integrase, and similar domains are present in prenyl transferases. we report the x-ray crystallographic structures of one diketo-acid and one keto-acid bound to crtm, which supports the mg(2+) ...201222662288
finding the smoking gun: protein tyrosine phosphatases as tools and targets of unicellular microorganisms and viruses.protein tyrosine phosphatases (ptps) are increasingly recognized as important effectors of host-pathogen interactions. since guan and dixon reported in 1990 that phosphatase yoph serves as an essential virulence determinant of yersinia, the field shifted significantly forward, and dozens of ptps were identified in various microorganisms and even in viruses. the discovery of extensive tyrosine signaling networks in non-metazoan organisms refuted the moth-eaten paradigm claiming that these organis ...201222360485
bacteria and their toxins tamed for immunotherapy.bacterial toxins share the ability to enter host cells to target various intracellular proteins and to modulate host immune responses. over the last 20 years, toxins and their mutated variants, as well as live attenuated bacteria, have been exploited for vaccination and immunotherapy of various infectious, malignant and autoimmune diseases. the ability of bordetella pertussis adenylate cyclase toxin to translocate its adenylate cyclase domain across the host cell membrane, as well as the pathway ...201222339216
the influences of natural zeolite (cliptinolite) on ammonia and biogenic amine formation by foodborne pathogen.the influence of natural zeolite on biogenic amines (bas) and ammonia (amn) production by eight common gram negative and positive foodborne pathogens (fbp) were investigated in histidine decarboxylase broth (hdb). presence of 1% zeolite in the hdb resulted in significantly higher amn production. histamine (his) production by gram positive bacteria was as low as 0.5 mg/l, whereas escherichia coli produced 18.96 mg/l of his. the use of zeolite also significantly suppressed his accumulation by e. c ...201222860594
design, structural and spectroscopic elucidation, and the in vitro biological activities of new diorganotin dithiocarbamates.the reaction of 2,2-dimethoxy-n-methylethyllamine or 2-methyl-1,3-dioxolane with cs(2) in alkaline media produced two novel dithiocarbamate salts. subsequent reactions with organotin halides yielded six new complexes: [snme(2){s(2)cnr(r(1))(2)}(2)] (1), [sn(n-bu)(2){s(2)cnr(r(1))(2)}(2)] (2), [snph(2){s(2)cnr(r(1))(2)}(2)] (3), [snme(2){s(2)cnr(r(2))(2)}(2)] (4), [sn(n-bu)(2){s(2)cnr(r(2))(2)}(2)] (5), [snph(2){s(2)cnr(r(2))(2)}(2)] (6), where r = methyl, r(1) = ch(2)ch(ome)(2), and r(2) = 2-met ...201223159807
immunomodulatory properties of enterococcus faecium jws 833 isolated from duck intestinal tract and suppression of listeria monocytogenes infection.the aim of this study was to evaluate the immunomodulatory properties of enterococcus faecium jws 833 (jws 833) isolated from duck intestine and compare them to those of lactobacillus rhamnosus gg (lgg), a proven immunity-enhancing probiotic. to investigate the immune-enhancing properties of jws 833, production of nitric oxide (no) and cytokines was measured in mouse peritoneal macrophages. in addition, a listeria monocytogenes challenge model was used in the assessment. it was found that heat-k ...201222709265
[foodborne infections in france].prevention and control measures implemented along the human food chain in recent decades have been largely successful, as shown by the dramatic decrease in the incidence of listeriosis and certain salmonella serotypes. however, foodborne illness continues to be a serious public health threat, due mainly to salmonella, listeria monocytogenes and campylobacter. infections caused by less frequent micro-organisms such as shigatoxin-producing e. coli are also a concern, because of their serious healt ...201224313018
effects of combined treatments of electron-beam irradiation and addition of leek (allium tuberosum) extract on reduction of pathogens in pork jerky.this study investigated the combined effect of electron-beam irradiation and addition of leek (allium tuberosum r.) extract on pork jerky inoculated with selected foodborne pathogens. prepared pork jerky samples (control and samples with 1.0% leek extract) were inoculated with pathogens and subsequently irradiated at 0, 0.5, 1, 2, 3, and 4 kgy doses. in comparison with the control, samples with 1.0% leek extract showed significant reduction in the numbers of escherichia coli, listeria monocytoge ...201223199493
modified-atmosphere packaging of hen table eggs: effects on pathogen and spoilage bacteria.as part of a more comprehensive research activity on the use of modified-atmosphere packaging for the improvement of quality and functional properties of table eggs, the effects of air, 100% co(2), and 100% o(2) packaging were also evaluated on the survival of experimentally inoculated pathogen bacteria (salmonella enteritidis, escherichia coli, and listeria monocytogenes) as well as on spoilage bacteria (total aerobic mesophilic bacteria) on table eggs during 30 d of storage at 4, 25, and 37°c ...201223155038
development of bacteria identification array to detect lactobacilli in thai fermented sausage.to improve the quality and safety of food products, there is a need in the food industry for a reliable method for simultaneously monitoring multiple bacterial strains. microarray technology is a high-throughput screening approach that can provide an alternative for bacteria detection. a total of 164 bacteria-specific probes were designed from 16s rrna gene sequences to target 12 bacteria species, including lactic acid bacteria and selected food pathogens. after fabrication onto aminosilane-coat ...201223022427
[zoonotic potential of raw cow's milk in the czech republic].to assess current microbiological risks of raw cow's milk. the presented work reports the prevalence of selected bacteria on czech dairy farms in 2010 and compares the results with a similar study in 2002.201222997775
glutamate decarboxylase-dependent acid resistance in orally acquired bacteria: function, distribution and biomedical implications of the gadbc operon.for successful colonization of the mammalian host, orally acquired bacteria must overcome the extreme acidic stress (ph < 2.5) encountered during transit through the host stomach. the glutamate-dependent acid resistance (gdar) system is by far the most potent acid resistance system in commensal and pathogenic escherichia coli, shigella flexneri, listeria monocytogenes and lactococcus lactis. gdar requires the activity of glutamate decarboxylase (gadb), an intracellular plp-dependent enzyme which ...201222995042
foodborne pathogens in in-line milk filters and associated on-farm risk factors in dairy farms authorized to produce and sell raw milk in northern italy.all dairy farms authorized to produce and sell raw milk in a province of northern italy were investigated to determine the presence of campylobacter spp., verocytotoxin-producing escherichia coli (vtec), listeria monocytogenes, and salmonella spp. in in-line milk filters and to assess their association with suspected risk factors on farms. a logistic regression model was used to analyze data collected describing the characteristics and management practices of 27 farms and the microbiological sta ...201222980010
effects of pressurization on some contamination flora in beef pate.pate meat is one of the food products at the top of the high-risk category since it is an undercooked product, which does not undergo any treatment to ensure its safety before consumption. in this study, pressurization, a nonthermal technology, was administered to the product to enhance its microbial safety for the 1st time. after being cooked, ground, and mixed with broth and spices, the pate meat was divided into 3 batches. their additional fat contents were adjusted to 9.11%, 25.00%, and 35.0 ...201222924817
the timing of ifnβ production affects early innate responses to listeria monocytogenes and determines the overall outcome of lethal infection.dendritic cells (dcs) and natural killer (nk) cells are essential components of the innate immunity and play a crucial role in the first phase of host defense against infections and tumors. listeria monocytogenes (lm) is an intracellular pathogen that colonizes the cytosol of eukaryotic cells. recent findings have shown lm specifically in splenic cd8a(+) dcs shortly after intravenous infection. we examined gene expression profiles of mouse dcs exposed to lm to elucidate the molecular mechanisms ...201222912878
effects of holdfast of laminaria japonica on listeria invasion on enterocyte-like caco-2 cells and no production of macrophage raw 264.7 cells.listeria monocytogenes (lm) causes food poisoning in humans mainly through consumption of ready-to-eat foods. immunocompromised persons are at the highest risk for infection. we investigated effects of crude soluble polysaccharides (sps) and ethanolic extract (ee) fractions of frond (kombu) and holdfast (ganiashi) parts of laminaria japonica on lm invasion into human enterocyte-like caco-2 cells and immune and/or inflammatory reactions of murine macrophage raw 264.7 cells. recovery and viscosity ...201222893519
listeria monocytogenes peritonitis: presentation, clinical features, treatment, and outcome.spontaneous bacterial peritonitis (sbp) is a serious complication in cirrhotic patients. gram (-) (e. coli, klebsiella pneumoniae), and gram (+) (streptococci, staphylococci) bacteria are most frequently cultured from patients'ascites. listeria monocytogenes (lm) is scarcely reported as a causative agent.201222834987
the activity of ferulic and gallic acids in biofilm prevention and control of pathogenic bacteria.the activity of two phenolic acids, gallic acid (ga) and ferulic acid (fa) at 1000 μg ml(-1), was evaluated on the prevention and control of biofilms formed by escherichia coli, pseudomonas aeruginosa, staphylococcus aureus and listeria monocytogenes. in addition, the effect of the two phenolic acids was tested on planktonic cell susceptibility, bacterial motility and adhesion. biofilm prevention and control were tested using a microtiter plate assay and the effect of the phenolic acids was asse ...201222823343
[mri findings of neonatal purulent meningitis caused by different pathogenic bacteria].to study the image characteristics of neonatal purulent meningitis caused by different pathogenic bacteria, using conventional magnetic resonance imaging (mri).201222809598
pulsed light treatment for the inactivation of selected pathogens and the shelf-life extension of beef and tuna carpaccio.the efficacy of pulsed light to improve the safety of carpaccio has been investigated. beef and tuna slices were superficially inoculated with approximately 3 log cfu/cm2 of listeria monocytogenes, escherichia coli, salmonella typhimurium and vibrio parahaemolyticus. fluences of 0.7, 2.1, 4.2, 8.4 and 11.9 j/cm2 were assayed. colour, sensory and shelf-life studies were carried out. treatments at 8.4 and 11.9 j/cm2 inactivated the selected pathogens approximately by 1 log cfu/cm2, although they m ...201222795799
nlrp6 negatively regulates innate immunity and host defence against bacterial pathogens.members of the intracellular nucleotide-binding and oligomerization domain (nod)-like receptor (nlr) family contribute to immune responses through activation of nuclear factor-κb (nf-κb), type i interferon and inflammasome signalling. mice lacking the nlr family member nlrp6 were recently shown to be susceptible to colitis and colorectal tumorigenesis, but the role of nlrp6 in microbial infections and the nature of the inflammatory signalling pathways regulated by nlrp6 remain unclear. here we s ...201222763455
construction of recombinant e. coli nissle 1917 (ecn) strains for the expression and secretion of defensins.the probiotic escherichia coli strain nissle 1917 (ecn) is one of the few probiotics licensed as a medication in several countries. best documented is its effectiveness in keeping patients suffering from ulcerative colitis (uc) in remission. this might be due to its ability to induce the production of human β-defensin 2 (hbd2) in a flagellin-dependent way in intestinal epithelial cells. in contrast to ulcerative colitis, for crohn's disease (cd) convincing evidence is lacking that ecn might be c ...201222748509
identification of volatile components of liverwort (porella cordaeana) extracts using gc/ms-spme and their antimicrobial activity.chemical constituents of liverwort (porella cordaeana) extracts have been identified using solid-phase microextraction-gas chromatography mass spectrometry (spme-gc/ms). the methanol, ethanol and ethyl acetate extracts were rich in terpenoids such as sesquiterpene hydrocarbons (53.12%, 51.68%, 23.16%), and monoterpene hydrocarbons (22.83%, 18.90%, 23.36%), respectively. the dominant compounds in the extracts were β-phellandrene (15.54%, 13.66%, 12.10%) and β-caryophyllene (10.72%, 8.29%, 7.79%, ...201222728370
[accelerated method for microbiological control of the safety of foodstuffs].the action and devices of rapid singlepath (merck kgaa) tests are considered. the singlepath tests are used for the accelerated detection of pathogenic bacteria (salmonella spp., campylobacter spp., listeria monocytogenes, e. coli o157) in the raw products and foodstuff. the tests as compared to conventional methods showed a high similarity of results in identifying salmonella in raw meat material. the immunochromatographic test requires no equipment and it is characterized by simple sample prep ...201222712333
microbiological profile of milk: impact of household practices.milk is susceptible to contamination by many microorganisms including microbial pathogens responsible for causing diseases. various processes including pasteurization, boiling or storage under refrigerated conditions are undertaken to minimize the microbial contamination of milk.201222684182
antimicrobial food packaging film based on the release of lae from evoh.the aim of this work was to develop antimicrobial films for active packaging applications containing the natural antimicrobial compound lae (lauramide arginine ethyl ester) in evoh copolymers with different mol % ethylene contents (i.e. evoh-29 and evoh-44). evoh-29 and evoh-44 films were made by casting and incorporating 0.25%, 1%, 5%, and 10% lae in the film forming solution (w/w with respect to polymer weight). previously, the minimum inhibitory concentration (mic) and the minimum bactericida ...201222640726
development of antimicrobial films for microbiological control of packaged salad.the aim of the present work was to characterize the antimicrobial efficiency of films consisting of pp/evoh structures with oregano essential oil and citral. both substances are known for their antimicrobial activity based on their interaction with the cell membrane. the films developed were used to pack minimally processed salads, combining modified atmosphere technology to extend shelf-life and active packaging technology to reduce possible microbiological risks. the antimicrobial activity of ...201222633535
estimates of enteric illness attributable to contact with animals and their environments in the united states.contact with animals and their environment is an important, and often preventable, route of transmission for enteric pathogens. this study estimated the annual burden of illness attributable to animal contact for 7 groups of pathogens: campylobacter species, cryptosporidium species, shiga toxin-producing escherichia coli (stec) o157, stec non-o157, listeria monocytogenes, nontyphoidal salmonella species, and yersinia enterocolitica.201222572672
high-intensity 405 nm light inactivation of listeria monocytogenes.the antimicrobial properties of light is an area of increasing interest. this study investigates the sensitivity of the significant foodborne pathogen listeria monocytogenes to selected wavelengths of visible light. results demonstrate that exposure to wavelength region 400-450 nm, at sufficiently high dose levels (750 j cm(-2)), induced complete inactivation of a 5 log(10) population. exposure to wavelengths longer than 450 nm did not cause significant inactivation. analysis of 10 nm bandwidths ...201222582879
multidrug-resistant bacterial infection in solid organ transplant recipients.the most frequent complication from infection after solid organ transplantation is bacterial infection. this complication is more frequent in organ transplantation involving the abdominal cavity, such as liver or pancreas transplantation, and less frequent in heart transplant recipients. the sources, clinical characteristics, antibiotic resistance and clinical outcomes vary according to the time of onset after transplantation. most bacterial infections during the first month post-transplantation ...201222542034
study on carvacrol and cinnamaldehyde polymeric films: mechanical properties, release kinetics and antibacterial and antibiofilm activities.polyethylene-co-vinylacetate (eva) films with different concentrations (3.5 wt% and 7 wt%) of essential oil constituents, carvacrol or cinnamaldehyde, were prepared and characterized by mechanical, antibacterial and antibiofilm properties. the incorporation of the compounds into copolymer films affected their elastic modulus, tensile stress and elongation at break. carvacrol and cinnamaldehyde act as plasticizers which reduce the intermolecular forces of polymer chains, thus improving the flexib ...201222555914
factors affecting quality and safety of fresh-cut produce.the quality of fresh-cut fruit and vegetable products includes a combination of attributes, such as appearance, texture, and flavor, as well as nutritional and safety aspects that determine their value to the consumer. nutritionally, fruit and vegetables represent a good source of vitamins, minerals, and dietary fiber, and fresh-cut produce satisfies consumer demand for freshly prepared, convenient, healthy food. however, fresh-cut produce deteriorates faster than corresponding intact produce, a ...201222530712
variability of rna quality extracted from biofilms of foodborne pathogens using different kits impacts mrna quantification by qpcr.the biofilm formation by foodborne pathogens is known to increase the problem related with surface disinfection procedure in the food processing environment and consequent transmission of these pathogens into the population. messenger rna has been increasingly used to understand the action and the consequences of disinfectants in the virulence on such biofilms. rna quality is an important requirement for any rna-based analysis since the quality can impair the mrna quantification. therefore, we e ...201222526569
bactericidal antibiotics do not appear to cause oxidative stress in listeria monocytogenes.oxidative stress can be an important contributor to the lethal effect of bactericidal antibiotics in some bacteria, such as escherichia coli and staphylococcus aureus. thus, despite the different target-specific actions of bactericidal antibiotics, they have a common mechanism leading to bacterial self-destruction by internal production of hydroxyl radicals. the purpose of the present study was to determine if a similar mechanism is involved in antibiotic killing of the infectious human pathogen ...201222504823
insight into the prevalence and distribution of microbial contamination to evaluate water management in the fresh produce processing industry.this study provided insight into the degree of microbial contamination in the processing chain of prepacked (bagged) lettuce in two belgian fresh-cut produce processing companies. the pathogens salmonella and listeria monocytogenes were not detected. total psychrotrophic aerobic bacterial counts (tpacs) in water samples, fresh produce, and environmental samples suggested that the tpac is not a good indicator of overall quality and best manufacturing practices during production and processing. be ...201222488054
microbiological surveillance of a bovine raw milk farm through multiplex real-time pcr.raw milk is increasingly appreciated by consumers but can be contaminated by a variety of zoonotic pathogens. therefore, preventive measures, such as on-farm hazard analysis critical control point (haccp) programs, must be applied to protect consumers. the aim of the present study was the comparison of a multiplex real-time polymerase chain reaction (pcr) assay with a culture-based approach in an on-farm quality assurance program for the detection of escherichia coli o157, salmonella spp., and l ...201222471929
[prokaryotic expression and identification of hpv16 e7 protein].hpv16 e7 fusion protein was expressed in e. coli bl21, and its applied value for hpv was evaluated. hpv16 e7 gene was amplified by pcr, and cloned into prokaryotic expression vector pgex6p-1. the recombinant plasmid was transformed into e. coli bl21, and hpv16 e7 fusion was expressed through iptg induction. the expressed product was analyzed by sds-page and western blot, subsequently purified according to glutathione sepharose 4b purification procedure. an indirect elisa with the purified fusion ...201222416350
nexilin is a dynamic component of listeria monocytogenes and enteropathogenic escherichia coli actin-rich structures.the bacterial pathogens listeria monocytogenes and enteropathogenic escherichia coli (epec) generate motile actin-rich structures (comet tails and pedestals) as part of their infectious processes. nexilin, an actin-associated protein and a component of focal adhesions, has been suggested to be involved in actin-based motility. to determine whether nexilin is commandeered during l. monocytogenes and epec infections, we infected cultured cells and found that nexilin is crucial for l. monocytogenes ...201222381134
changing epidemiology of bacteremia in infants aged 1 week to 3 months.bacteremia in young infants has remained an important ongoing concern for decades. despite changes in prenatal screening and infant immunizations, the current epidemiology of this problem has received little attention.201222371459
a selected probiotic strain of lactobacillus fermentum cm33 isolated from breast-fed infants as a potential source of β-galactosidase for prebiotic oligosaccharide synthesis.lactic acid bacteria from healthy breast-fed infants were isolated and screened for β-galactosidase production in mrs broth. among 49 isolates that exhibited the yellow clear zone on mrs agar supplemented with bromocresol blue, the isolate cm33 was selected as being the highest β-galactosidase producer and was identified as lactobacillus fermentum based on its morphological characteristics and 16s rdna nucleotide sequence. l. fermentum cm33 exhibited a good survival rate under the simulated stom ...201222367946
recombinant expression of a putative amidase cloned from the genome of listeria monocytogenes that lyses the bacterium and its monolayer in conjunction with a protease.listeria monocytogenes is a gram-positive, non-spore forming, catalase-positive rod that is a major bacterial food-borne disease agent associated with uncooked meats, including poultry, uncooked vegetables, soft cheeses, and unpasteurized milk. the bacterium may be carried by animals without signs of disease, can replicate at refrigeration temperatures, and is frequently associated with biofilms. there is a need to discover innovative pathogen intervention technologies for this bacterium. conseq ...201226781731
fructooligosaccharides metabolism and effect on bacteriocin production in lactobacillus strains isolated from ensiled corn and molasses.fructo- (fos) and galacto-oligosaccharides have been used to promote the growth of probiotics, mainly those from lactobacillus genus. however, only few reports have evaluated the effect of prebiotics on bacteriocins activity and production. in this work, we characterized the effect of fos supplementation on the growth, lactic and acetic acids production, and antimicrobial activity of crude extracts obtained from lactobacillus strains isolated from ensiled corn and molasses. seven out of 28 isola ...201222342961
development of a paper-based analytical device for colorimetric detection of select foodborne pathogens.foodborne pathogens are a major public health threat and financial burden for the food industry, individuals, and society, with an estimated 76 million cases of food-related illness occurring in the united states alone each year. three of the most important causative bacterial agents of foodborne diseases are pathogenic strains of escherichia coli , salmonella spp., and listeria monocytogenes , due to the severity and frequency of illness and disproportionally high number of fatalities. their co ...201222320200
cloning and heterologous expression of a bacteriocin sakacin p from lactobacillus sakei in escherichia coli.sakacin p, a bacteriocin from lactobacillus sakei, shows strong activity against food-borne pathogens such as listeria monocytogenes. in l. sakei, the structural gene (sppa) encoding sakacin p is controlled by a strict regulatory mechanism, and the quantity of secreted sakacin p is limited. in this study, the sppa gene was synthesized by splicing overlap extension pcr and cloned into escherichia coli. after the induction with isopropyl-β-d-thiogalactopyranoside, the recombinant sakacin p was suc ...201222290644
antimicrobial activity and chemical composition of essential oil from the seeds of artemisia aucheri boiss.artemisia aerial parts are well known for antimicrobial activities including anti malaria.201224624145
[analysis of international programs of the hygienic monitoring of foodborne pathogens].the article gives an overview of information international networks of monitoring the microbiological food safety in the world. the most famous are the european network of food safety (efsn), global network for disease surveillance, foodborne (gfn), international network for the determination of molecular subtypes of bacterial food patogens (pulsenet), international network of food safety (infosan). the network efsn is engaged development of information database and of warning systems to assess ...201223530431
isolation of pediococcus acidilactici kp10 with ability to secrete bacteriocin-like inhibitory substance from milk products for applications in food industry.lactic acid bacteria (lab) can be isolated from traditional milk products. lab that secrete substances that inhibit pathogenic bacteria and are resistant to acid, bile, and pepsin but not vancomycin may have potential in food applications.201223153191
antimicrobial impact of the components of essential oil of litsea cubeba from taiwan and antimicrobial activity of the oil in food systems.using natural additives to preserve foods has become popular due to consumer demands for nature and safety. antimicrobial activity is one of the most important properties in many plant essential oils (eos). the antimicrobial activity of the essential oil of litsea cubeba (lc-eo) from taiwan and the antimicrobial impact of individual volatile components in the oil on pathogens or spoilage microorganisms: vibrio parahaemolyticus, listeria monocytogenes, lactobacillus plantarum, and hansenula anoma ...201222459760
[lactobacillus plantarum and phosprenyl competitively distinguish ovalbumin oligomannoside n-glycan].demonstration of the ability of native products of lactobacteria to bind mannose containing n-glycans as a display of one of the mechanisms of probiotic behavior of these symbiont microorganism in gastrointestinal tract (git).201222308726
characterization of a reuterin-producing lactobacillus reuteri bpl-36 strain isolated from human infant fecal sample.a reuterin (3-hydroxypropinaldehyde, 3-hpa)-producing isolate from a human infant fecal sample was identified as lactobacillus reuteri bpl-36 strain. the organism displayed a broad-spectrum antimicrobial activity. the gene (gdh) encoding a glycerol dehydratase subunit was detected by pcr, thus confirming its reuterin-producing ability. reuterin concentration of 89.63 mm/ml was obtained in the mrs-glycerol medium after 16 h of incubation at 37 °c. the reuterin concentration required to inhibit th ...201226782041
lactobacillus sakei crl1862 improves safety and protein hydrolysis in meat systems.the capacity of lactobacillus sakei crl1862 to prevent the growth of pathogens and its ability to degrade sarcoplasmic and myofibrillar proteins in pork meat systems was evaluated. in addition, basic safety aspects of lact. sakei crl1862 such as production of biogenic amines and antibiotic susceptibility were addressed.201222963007
in vitro evaluation of the probiotic potential of bacteriocin producer lactobacillus sakei 1.lactobacillus sakei 1 is a food isolate that produces a heat-stable antimicrobial peptide (sakacin 1, a class iia bacteriocin) inhibitory to the opportunistic pathogen listeria monocytogenes. bacterial isolates with antimicrobial activity may be useful for food biopreservation and also for developing probiotics. to evaluate the probiotic potential of l. sakei 1, it was tested for (i) in vitro gastric resistance (with synthetic gastric juice adjusted to ph 2.0, 2.5, or 3.0); (ii) survival and bac ...201222691476
effect of brine marination on survival and growth of spoilage and pathogenic bacteria during processing and subsequent storage of ready-to-eat shrimp (pandalus borealis).the effect of brine marination at chill temperatures on survival and growth of spoilage and pathogenic bacteria during processing and subsequent storage of ready-to-eat cold water shrimp was studied. survival and growth of lactobacillus sakei, listeria monocytogenes, salmonella, staphylococcus aureus and vibrio parahaemolyticus were examined. the effect of brine composition and ph was determined in 12 screening experiments without addition of shrimp. sixteen challenge tests with shrimp were then ...201222564452
lacticin lc14, a new bacteriocin produced by lactococcus lactis bmg6.14: isolation, purification and partial characterization.a new bacteriocin, lacticin lc14, produced by lactococcus lactis bmg6.14, was isolated and characterized. it was purified to homogeneity from overnight broth culture by ammonium sulfate precipitation, sep-pak chromatography, and two steps of reversed-phase hplc. lacticin lc14 showed bactericidal-type antimicrobial activity against several lactic acid bacteria and pathogenic strains including listeria monocytogenes. it was inactivated by proteinase k and pronase e, but was resistant to papain, ly ...201222697129
therapeutic live vaccines as a potential anticancer strategy.the design of efficient cancer treatments is one of the major challenges of medical science. therapeutic vaccines of cancer have been emerged as an attractive approach for their capacity of breaking the immune tolerance and invoking long-term immune response targeting cancer cells without autoimmunity. an efficient antigen delivery system is the key issue of developing an effective cancer vaccine. in this regard, live vaccination strategies including various live bacterial and viral vectors have ...201222610886
antilisterial activity of nisin-like bacteriocin-producing lactococcus lactis subsp. lactis isolated from traditional sardinian dairy products.with the aim of selecting lab strains with antilisterial activity to be used as protective cultures to enhance the safety of dairy products, the antimicrobial properties of 117 lactococcus lactis subsp. lactis isolated from artisanal sardinian dairy products were evaluated, and six strains were found to produce bacteriocin-like substances. the capacity of these strains to antagonize listeria monocytogenes during cocultivation in skimmed milk was evaluated, showing a reduction of l. monocytogenes ...201222536018
biologic therapy and risk of infection.biologic compounds are being used more frequently to treat a multitude of systemic inflammatory conditions. these novel compounds are composed of antibodies or other peptides that act through one of three mechanisms: inhibiting inflammatory cytokine signaling (typically tumor necrosis factor or tnf), inhibiting t-cell activation, or depleting b-cells. the increase in use and ever expanding list of new immune modulating therapies make knowledge of the infectious complications associated with immu ...201222491803
in vitro and in vivo characterization of dna delivery using recombinant lactococcus lactis expressing a mutated form of l. monocytogenes internalin a.the use of food-grade lactic acid bacteria (lab) as dna delivery vehicles represents an attractive strategy to deliver dna vaccines at the mucosal surfaces as they are generally regarded as safe (gras). we previously showed that either native lactococcus lactis (ll) or recombinant invasive ll expressing fibronectin binding protein a of staphylococcus aureus (ll-fnbpa+) or internalin a of listeria monocytogenes (ll-inla+), were able to deliver and trigger dna expression by epithelial cells, eithe ...201223253484
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