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isolation and identification of quercetin degrading bacteria from human fecal microbes.quercetin has a wide range of biological properties. the gut microflora can often modulate its biological activity and their potential health effects. there still is a lack of information about gut bacteria involving in this process. the strains of gut microbes from human feces that can transform quercetin were isolated and identified by in vitro fermentation. the results showed that escherichia coli, stretococcus lutetiensis, lactobacillus acidophilus, weissella confusa, enterococcus gilvus, cl ...201424594786
short- and medium-chain fatty acids as a feed supplement for weaning and nursery pigs.the effect of supplementing piglet diets with acidifiers containing the short-chain fatty acids - scfa (propionic c3 and formic) together with medium-chain fatty acids -- mcfa (caprylic c8 and capric c10) on performance, nutrient apparent digestibility, intestinal microflora and small intestine structure was investigated. the study was performed on 326 piglets allocated to 5 experimental groups. they were fed a standard diet (group i - control) or a standard diet supplemented with 0.5% propionic ...201324597298
draft genome sequence of clostridium perfringens strain jjc, a highly efficient hydrogen producer isolated from landfill leachate sludge.clostridium perfringens strain jjc is an effective biohydrogen and biochemical producer that was isolated from landfill leachate sludge. here, we present the assembly and annotation of its genome, which may provide further insights into the gene interactions involved in efficient biohydrogen production.201424604637
antibacterial and antioxidant activities in extracts of fully grown cladodes of 8 cultivars of cactus pear.the antimicrobial and antioxidant activities of some cultivars of the nopal cactus have not been determined. in this study, 8 cultivars of nopal cacti from mexico were assayed for phenolic content, antioxidant activities, and antimicrobial activities against campylobacter jejuni, vibrio cholera, and clostridium perfringens. plant material was washed, dried, and macerated in methanol. minimum bactericidal concentrations (mbcs) were determined using the broth microdilution method. antioxidant acti ...201424621296
identification of a key residue for oligomerisation and pore-formation of clostridium perfringens netb.necrotic enteritis toxin b (netb) is a β-pore-forming toxin produced by clostridium perfringens and has been identified as a key virulence factor in the pathogenesis of avian necrotic enteritis, a disease causing significant economic damage to the poultry industry worldwide. in this study, site-directed mutagenesis was used to identify amino acids that play a role in netb oligomerisation and pore-formation. netb k41h showed significantly reduced toxicity towards lmh cells and human red blood cel ...201424625763
fluorescence in situ hybridization investigation of potentially pathogenic bacteria involved in neonatal porcine diarrhea.neonatal diarrhea is a multifactorial condition commonly present on pig farms and leads to economic losses due to increased morbidity and mortality of piglets. immature immune system and lack of fully established microbiota at birth predispose neonatal piglets to infection with enteric pathogens. the microorganisms that for decades have been associated with enteritis and diarrhea in suckling piglets are: rotavirus a, coronavirus, enterotoxigenic escherichia coli (etec), clostridium perfringens t ...201424628856
modulation of micrornas in two genetically disparate chicken lines showing different necrotic enteritis disease susceptibility.micrornas (mirna) play a critical role in post-transcriptional regulation by influencing the 3'-utr of target genes. using two inbred white leghorn chicken lines, line 6.3 and line 7.2 showing marek's disease-resistant and -susceptible phenotypes, respectively, we used small rna high-throughput sequencing (hts) to investigate whether mirnas are differently expressed in these two chicken lines after inducing necrotic enteritis (ne). the 12 mirnas, selected from the most down-regulated or up-regul ...201424629767
membrane vesicles of clostridium perfringens type a strains induce innate and adaptive immunity.vesicle shedding from bacteria is a universal process in most gram-negative bacteria and a few gram-positive bacteria. in this report, we isolate extracellular membrane vesicles (mvs) from the supernatants of gram-positive pathogen clostridium perfringens (c. perfringens). we demonstrated vesicle production in a variety of virulent and nonvirulent type a strains. mvs did not contain alpha-toxin and netb toxin demonstrated by negative reaction to specific antibody and absence of specific proteins ...201424631214
clostridium perfringens type a enterotoxin damages the rabbit colon.clostridium perfringens enterotoxin causes the gastrointestinal (gi) symptoms of c. perfringens type a food poisoning and cpe-associated non-food-borne human gi diseases. it is well established that cpe induces fluid accumulation and severe tissue damage in ligated small intestinal loops of rabbits and other animals. however, a previous study had also reported that cpe binds to rabbit colonic cells yet does not significantly affect rabbit colonic loops. to the contrary, the current study determi ...201424643537
molecular analysis of infant fecal microbiota in an asian at-risk cohort-correlates with infant and childhood eczema.studies have suggested that selective microbial targets prevail in the fecal microbiota of infants with eczema. this study evaluated the composition of fecal microbiota of infants who developed eczema in the first 5 years of life and compared these with those of healthy controls.201424650346
evaluation of a multiplex pcr assay for simultaneous detection of bacterial and viral enteropathogens in stool samples of paediatric patients.we evaluated a multiplex pcr assay, the seeplex diarrhoea ace detection, that simultaneously detects 15 enteric pathogens, including salmonella spp., shigella spp., vibrio spp., toxin b producer clostridium difficile, campylobacter spp., clostridium perfringens, yersinia enterocolitica, aeromonas spp., escherichia coli o157:h7, verocytotoxin-producing escherichia coli, adenovirus, group a rotavirus, norovirus gi and gii, and astrovirus. we compared this assay with clinical methods routinely used ...201424656922
synergistic antibacterial effect of honey and herba ocimi basilici against some bacterial pathogens.to evaluate the antibacterial activity of the combination of different honey brands and methanolic fraction of herba ocimi basilici using agar well diffusion assay.201324660616
empathy dolls: are they a source of cross-contamination between patients?following a cluster of two patients with identical strains of clostridium perfringens prosthetic joint infections on an ortho-geriatric ward in a teaching hospital in england, investigations were conducted into infection control practices. it emerged that empathy dolls were being used to help alleviate agitation in dementia patients; this had been introduced without consultation with the infection prevention and control team. environmental testing of the doll pre and post laundry at different te ...201424661788
bec, a novel enterotoxin of clostridium perfringens found in human clinical isolates from acute gastroenteritis outbreaks.clostridium perfringens is a causative agent of food-borne gastroenteritis for which c. perfringens enterotoxin (cpe) has been considered an essential factor. recently, we experienced two outbreaks of food-borne gastroenteritis in which non-cpe producers of c. perfringens were strongly suspected to be the cause. here, we report a novel enterotoxin produced by c. perfringens isolates, bec (binary enterotoxin of c. perfringens). culture supernatants of the c. perfringens strains showed fluid-accum ...201424664508
production of human metabolites by gastrointestinal bacteria as a potential source of post-mortem alteration of antemortem drug/metabolite concentrations.previous studies have demonstrated that bacterial species are capable of transforming complex chemical substances. several of these species, native to the human gastrointestinal tract, are active in postmortem decomposition. they have potential to cause biotransformations affecting compound-to-metabolite ratios within the human body, especially after death. investigation of postmortem effects could supply valuable information, especially concerning compound identification and confirmation. the p ...201524665046
x-ray structure of a novel endolysin encoded by episomal phage phism101 of clostridium perfringens.gram-positive bacteria possess a thick cell wall composed of a mesh polymer of peptidoglycans, which provides physical protection. endolysins encoded by phages infecting bacteria can hydrolyse peptidoglycans in the bacterial cell wall, killing the host bacteria immediately. the endolysin (psm) encoded by episomal phage phism101 of enterotoxigenic clostridium perfringens type a strain sm101 exhibits potent lytic activity towards most strains of clostridium perfringens. psm has an n-terminal catal ...201424674022
clostridium perfringens challenge and dietary fat type affect broiler chicken performance and fermentation in the gastrointestinal tract.the aim of the present work was to examine how different fats commonly used in the feed industry affect broiler performance, nutrient digestibility and microbial fermentation in the gastrointestinal tract of broiler chickens challenged with virulent clostridium perfringens strains. two experiments were carried out, each including 480-day-old male broilers (ross 308), which were randomly distributed to eight experimental groups using six replicate pens per treatment and 10 birds per pen. in exper ...201424674938
utility of the clostridial site-specific recombinase tnpx to clone toxic-product-encoding genes and selectively remove genomic dna fragments.tnpx is a site-specific recombinase responsible for the excision and insertion of the transposons tn4451 and tn4453 in clostridium perfringens and clostridium difficile, respectively. here, we exploit phenotypic features of tnpx to facilitate genetic mutagenesis and complementation studies. genetic manipulation of bacteria often relies on the use of antibiotic resistance genes; however, a limited number are available for use in the clostridia. the ability of tnpx to recognize and excise specific ...201424682304
[spondylodiscitis due to clostridium perfringens]. 201424684888
heterologous protection against alpha toxins of clostridium perfringens and staphylococcus aureus induced by binding domain recombinant chimeric protein.clostridium perfringens and staphylococcus aureus are the two important bacteria frequently associated with majority of the soft tissue infections. the severity and progression of the diseases caused by these pathogens are attributed primarily to the alpha toxins they produce. previously, we synthesized a non-toxic chimeric molecule r-αcs encompassing the binding domains of c. perfringens and s. aureus alpha toxins and demonstrated that the r-αcs hyperimmune polysera reacts with both the native ...201424699467
perfrin, a novel bacteriocin associated with netb positive clostridium perfringens strains from broilers with necrotic enteritis.necrotic enteritis in broiler chickens is associated with netb positive clostridium perfringens type a strains. it is known that c. perfringens strains isolated from outbreaks of necrotic enteritis are more capable of secreting factors inhibiting growth of other c. perfringens strains than strains isolated from the gut of healthy chickens. this characteristic could lead to extensive and selective presence of a strain that contains the genetic make-up enabling to secrete toxins that cause gut les ...201424708344
clostridium perfringens epsilon toxin mutant y30a-y196a as a recombinant vaccine candidate against enterotoxemia.epsilon toxin (etx) is a β-pore-forming toxin produced by clostridium perfringens toxinotypes b and d and plays a key role in the pathogenesis of enterotoxemia, a severe, often fatal disease of ruminants that causes significant economic losses to the farming industry worldwide. this study aimed to determine the potential of a site-directed mutant of etx (y30a-y196a) to be exploited as a recombinant vaccine against enterotoxemia. replacement of y30 and y196 with alanine generated a stable variant ...201424709588
necrotic enteritis in chickens: development of a straightforward disease model system.the interaction between eimeria species and clostridium perfringens was investigated in two different necrotic enteritis (ne) models: 120-day-old broilers were used in two separate experiments consisting of six groups (n=10) each. besides controls, chickens were infected with coccidia on study day (sd) 18 (eimeria maxima and eimeria acervulina (experiment 1) or eimeria tenella and eimeria brunetti (experiment 2) and/or a netb toxin positive c perfringens strain (both experiments: sd 14 or sd 22, ...201424714053
complication of invasive molar pregnancy with clostridium perfringens sepsis.clostridium perfringens (cp) is an anaerobic, gram-positive bacillus associated with malignant diseases and near-term pregnancies. the necrotic tissue that results from these disease processes fuels the proliferation of cp, leading to gas gangrene and subsequently sepsis. herein, we report a case of a 41-year-old female patient with a history of invasive molar pregnancy that was further complicated with a cp infection. although past research has shown a link between clostridium infection and cho ...201424716030
haemorhagic enterotoxemia by clostridium perfringens type c and type a in silver foxes.type c and type a of c. perfringens were detected in the seat of natural infections in silver foxes characterized by symptoms of haemorrhagic enterotoxemia. in all of the dead foxes characteristic changes were noted in the small intestine and parenchymatous organs. the production of alpha and beta toxins by isolated bacteria was confirmed by the bioassay using white mice and by pcr. the results of the drug sensitivity testing showed that isolated strains were highly susceptible to amoxicillin wi ...201424724490
application of a real-time pcr method for salmonella spp., escherichia coli, staphylococcus aureus and clostridium perfringens detection in water samples.the diagnostic assessment of water sanitary state is based mainly on the cultivation of bacteria retained on membrane filters. however classical microbiology methods have a lot of disadvantages. more and more frequently, rapid detection and identification of pathogens present in water is based on molecular biology techniques. the aim of this study was to determine the effectiveness and usefulness of a real-time pcr method, when compared to the recommended bacteria culture method, in diagnostics ...201324730140
lrp1 is a receptor for clostridium perfringens tpel toxin indicating a two-receptor model of clostridial glycosylating toxins.large glycosylating toxins are major virulence factors of various species of pathogenic clostridia. prototypes are clostridium difficile toxins a and b, which cause antibiotics-associated diarrhea and pseudomembranous colitis. the current model of the toxins' action suggests that receptor binding is mediated by a c-terminal domain of combined repetitive oligopeptides (crop). this model is challenged by the glycosylating clostridium perfringens large cytotoxin (tpel toxin) that is devoid of the c ...201424737893
liver abscess caused by clostridium perfringens. 201424739619
retinal microvascular damage and vasogenic edema produced by clostridium perfringens type d epsilon toxin in rats.when the brain is exposed to large circulating levels of clostridium perfringens type d epsilon toxin (ext), microvascular damage with resulting severe, generalized, vasogenic edema seems to be principally responsible for the ensuing acute, and frequently fatal, neurologic disorder. however, although the blood-retinal barrier resembles in many respects the blood-brain barrier, retinal changes in livestock with acute epsilon intoxication have not, to the authors' knowledge, been previously report ...201424741023
relationship between gastrointestinal dysbiosis and clostridium botulinum in dairy cows.the gastrointestinal tract is a balanced ecosystem that can get out of balance and predisposed to clostridial diseases or other pathological conditions. the objective of the present study was to evaluate the gut microbiota in dairy cows suffering from chronic botulism. cows were investigated for clostridium (c.) botulinum in faeces and rumen fluids. in order to study the relationship between botulism and gastrointestinal microbiota, faeces and rumen fluid were tested for bacterial composition us ...201424747040
'core species' in three sources of indoor air belonging to the human micro-environment to the exclusion of outdoor air.although we spend the majority of our lives indoors, the airborne microbial content of enclosed spaces still remains inadequately described. the objective of this study was to characterize the bacterial diversity of indoor air in three different enclosed spaces with three levels of occupancy, and, in particular, to highlight the 'core' species, the opportunistic pathogens and their origins. our findings provide an overall description of bacterial diversity in these indoor environments. data gath ...201424747243
the impact of major earthquakes and subsequent sewage discharges on the microbial quality of water and sediments in an urban river.a series of large earthquakes struck the city of christchurch, new zealand in 2010-2011. major damage sustained by the sewerage infrastructure required direct discharge of up to 38,000 m(3)/day of raw sewage into the avon river of christchurch for approximately six months. this allowed evaluation of the relationship between concentrations of indicator microorganisms (escherichia coli, clostridium perfringens and f-rna phage) and pathogens (campylobacter, giardia and cryptosporidium) in recreatio ...201424747258
hypermotility in clostridium perfringens strain sm101 is due to spontaneous mutations in genes linked to cell division.clostridium perfringens is a gram-positive anaerobic pathogen of humans and animals. although they lack flagella, c. perfringens bacteria can still migrate across surfaces using a type of gliding motility that involves the formation of filaments of bacteria lined up in an end-to-end conformation. in strain sm101, hypermotile variants are often found arising from the edges of colonies on agar plates. hypermotile cells are longer than wild-type cells, and video microscopy of their gliding motility ...201424748614
gas-forming liver abscess associated with rapid hemolysis in a diabetic patient.we experienced a case of liver abscess due to clostridium perfringens (cp) complicated with massive hemolysis and rapid death in an adequately controlled type 2 diabetic patient. the patient died 6 h after his first visit to the hospital. cp was later detected in a blood culture. we searched for case reports of cp septicemia and found 124 cases. fifty patients survived, and 74 died. of the 30 patients with liver abscess, only 3 cases survived following treatment with emergency surgical drainage. ...201424748935
the effect of new neonatal porcine diarrhoea syndrome (nnpds) on average daily gain and mortality in 4 danish pig herds.the study evaluated the effect of new neonatal porcine diarrhoea syndrome (nnpds) on average daily gain (adg) and mortality and described the clinical manifestations in four herds suffering from the syndrome. nnpds is a diarrhoeic syndrome affecting piglets within the first week of life, which is not caused by enterotoxigenic escherichia coli (etec), clostridium perfringens (c. perfringens) type a/c, clostridium difficile (c. difficile), rotavirus a, coronavirus, cystoisospora suis, strongyloide ...201424755093
hepatic emphysema associated with ultrasound-guided liver biopsy in a dog.an eleven-year-old chinese crested powder puff dog presented with polydipsia/polyuria, inappetence, diarrhea and vomiting underwent an ultrasound-guided percutaneous liver biopsy. two days post-biopsy the clinical condition of the dog acutely deteriorated with fever, dyspnea, ataxia and subcutaneous emphysema. radiographs and ultrasound showed focal severe hepatic emphysema in the region of the previous liver biopsy. post-mortem examination revealed chronic hepatitis with dissecting fibrosis, ac ...201424758285
antibiotic resistance of clostridium perfringens isolates from broiler chickens in egypt.the use of antibiotic feed additives in broiler chickens results in a high prevalence of resistance among their enteric bacteria, with a consequent emergence of antibiotic resistance in zoonotic enteropathogens. despite growing concerns about the emergence of antibiotic-resistant strains, which show varying prevalences in different geographic regions, little work has been done to investigate this issue in the middle east. this study provides insight into one of the world's most common and financ ...201324761735
towards an understanding of the role of clostridium perfringens toxins in human and animal disease.clostridium perfringens uses its arsenal of >16 toxins to cause histotoxic and intestinal infections in humans and animals. it has been unclear why this bacterium produces so many different toxins, especially since many target the plasma membrane of host cells. however, it is now established that c. perfringens uses chromosomally encoded alpha toxin (a phospholipase c) and perfringolysin o (a pore-forming toxin) during histotoxic infections. in contrast, this bacterium causes intestinal disease ...201424762309
fulminating septic shock from clostridium perfringens in an early breast cancer patient with severe myalgia after docetaxel treatment.anaerobic bacteraemia could be a life-threatening condition in neutropenic patients receiving chemotherapy. taxane therapy is associated with necrotising inflammation of the caecum (named also typhlitis) that could be a potential source for bacteraemia. we report the case of a sudden onset of septic shock by clostridium perfringens in a young patient treated with docetaxel as adjuvant chemotherapy for early breast cancer. a mini-review of the literature has been performed.201124765295
beta2 toxin is not involved in in vitro cell cytotoxicity caused by human and porcine cpb2-harbouring clostridium perfringens.clostridium perfringens is a common cause of intestinal disease in animals and humans. its pathogenicity is attributed to the toxins it can produce, including the beta2 toxin. the presence of cpb2, the gene encoding the beta2 toxin, has been associated with diarrhoea in neonatal piglets and humans. however, the exact role of the beta2 toxin in the development of diarrhoea is still unknown. in this study we investigated the level of cytotoxicity to porcine ipi-21 and human caco-2 cell-lines cause ...201424768003
synergistic effects of clostridium perfringens enterotoxin and beta toxin in rabbit small intestinal loops.the ability of clostridium perfringens type c to cause human enteritis necroticans (en) is attributed to beta toxin (cpb). however, many en strains also express c. perfringens enterotoxin (cpe), suggesting that cpe could be another contributor to en. supporting this possibility, lysate supernatants from modified duncan-strong sporulation (mds) medium cultures of three cpe-positive type c en strains caused enteropathogenic effects in rabbit small intestinal loops, which is significant since cpe i ...201424778117
pathogen exclusion properties of canine probiotics are influenced by the growth media and physical treatments simulating industrial processes.manufacturing process used in preparation of probiotic products may alter beneficial properties of probiotics. the effect of different growth media and inactivation methods on the protective properties of canine-originated probiotic bacteria against adhesion of canine enteropathogens was investigated.201424779583
structure and stability of an azoreductase with an fad cofactor from the strict anaerobe clostridium perfringens.azoreductase enzymes present in many microorganisms exhibit the ability to reduce azo dyes, an abundant industrial pollutant, to produce carcinogenic metabolites that threaten human health. all biochemically-characterized azoreductases, around 30 to date, have been isolated from aerobic bacteria, except for azoc, the azoreductase of clostridium perfringens, which is from a strictly anaerobic bacterium. azoc is a recently biochemically-characterized azoreductase. the lack of structural informatio ...201424779771
essential oil composition and antimicrobial activity of angelica archangelica l. (apiaceae) roots.in this paper, the chemical composition and the antimicrobial activity of the essential oil of angelica archangelica l. (apiaceae) roots from central italy were analyzed. the major constituents of the oil were α-pinene (21.3%), δ-3-carene (16.5%), limonene (16.4%) and α-phellandrene (8.7%). the oil shows a good antimicrobial activity against clostridium difficile, clostridium perfringens, enterococcus faecalis, eubacterium limosum, peptostreptococcus anaerobius, and candida albicans with minimum ...201424788027
analysis of genetic similarities between clostridium perfringens isolates isolated from patients with gas gangrene and from hospital environment conducted with the use of the pfge method.the objective of the study was to perform a comparative analysis of genetic similarity, with the use of pulsed field gel electrophoresis (pfge), of clostridium perfringens isolates originating from patients with gas gangrene and from the hospital environment. the study encompassed two patients with a clinical and microbiological diagnosis of gas gangrene, who were hospitalized in one of the hospitals of the małopolska province in the time period between 31st march 2012 and 18th may 2012. clostri ...201424791817
effects of in ovo injection with selenium on immune and antioxidant responses during experimental necrotic enteritis in broiler chickens.this study was conducted to investigate the effects of in ovo injection of se on modulating the immune system and antioxidant responses in broiler chickens with experimental necrotic enteritis. broiler eggs were injected at 18 d of embryo age with either 100 μl of pbs alone or sodium selenite (na2seo3) in pbs, providing 0 (ss0), 10 (ss10), or 20 (ss20) μg of se/egg. at 14 d posthatch, pbs-treated and uninfected chickens were kept as the control group, whereas the remaining chickens were orally i ...201424795303
eutrophication and bacterial pathogens as risk factors for avian botulism outbreaks in wetlands receiving effluents from urban wastewater treatment plants.due to the scarcity of water resources in the "mancha húmeda" biosphere reserve, the use of treated wastewater has been proposed as a solution for the conservation of natural threatened floodplain wetlands. in addition, wastewater treatment plants of many villages pour their effluent into nearby natural lakes. we hypothesized that certain avian pathogens present in wastewater may cause avian mortalities which would trigger avian botulism outbreaks. with the aim of testing our hypothesis, 24 loca ...201424795377
biofilm formation of clostridium perfringens and its exposure to low-dose antimicrobials.clostridium perfringens is an opportunistic pathogen that can cause food poisoning in humans and various enterotoxemia in animal species. very little is known on the biofilm of c. perfringens and its exposure to subminimal inhibitory concentrations of antimicrobials. this study was undertaken to address these issues. most of the c. perfringens human and animal isolates tested in this study were able to form biofilm (230/277). porcine clinical isolates formed significantly more biofilm than the p ...201424795711
perfringolysin o structure and mechanism of pore formation as a paradigm for cholesterol-dependent cytolysins.cholesterol-dependent cytolysins (cdcs) constitute a family of pore forming toxins secreted by gram-positive bacteria. these toxins form transmembrane pores by inserting a large β-barrel into cholesterol-containing membrane bilayers. binding of water-soluble cdcs to the membrane triggers the formation of oligomers containing 35-50 monomers. the coordinated insertion of more than seventy β-hairpins into the membrane requires multiple structural conformational changes. perfringolysin o (pfo), secr ...201424798008
use of essential oils and extracts from spices in meat protection.the hydro distillation method was used in this study to get essential oils (eos) from cumin (cuminum cyminum l.), clove (eugenia caryohyllata) and elecampane (inula helenium l.) and the co-hydro distillation method (addition of fatty acid ethyl esters as extraction cosolvents) to get functional extracts (efs). the mic (minimum inhibitory concentration) and the mbc (minimum bactericidal concentration) were determined on five pathogenic strains (escherichia coli o157:h7, salmonella, listeria monoc ...201124803704
effects of melatonin on oxidative stress, and resistance to bacterial, parasitic, and viral infections: a review.melatonin, a hormone secreted by the pineal gland, works directly and indirectly as a free radical scavenger. its other physiological or pharmacological activities could be dependent or independent of receptors located in different cells, organs, and tissues. in addition to its role in promoting sleep and circadian rhythms regulation, it has important immunomodulatory, antioxidant, and neuroprotective effects suggesting that this indole must be considered as a therapeutic alternative against inf ...201424811367
antibacterial activity against clostridium genus and antiradical activity of the essential oils from different origin.in the present study, the antimicrobial and antiradical activities of 15 essential oils were investigated. the antimicrobial activities were determined by using agar disc diffusion and broth microdilution methods against clostridium genus and antioxidant properties of essential oils by testing their scavenging effect on dpph radicals activities. we determined the antibacterial activity of clostridium butyricum, clostridium hystoliticum, clostridium intestinale, clostridium perfringens and clostr ...201424813985
structural basis of sialidase in complex with geranylated flavonoids as potent natural inhibitors.sialidase catalyzes the removal of a terminal sialic acid from glycoconjugates and plays a pivotal role in nutrition, cellular interactions and pathogenesis mediating various infectious diseases including cholera, influenza and sepsis. an array of antiviral sialidase agents have been developed and are commercially available, such as zanamivir and oseltamivir for treating influenza. however, the development of bacterial sialidase inhibitors has been much less successful. here, natural polyphenoli ...201424816104
bovicin hj50-like lantibiotics, a novel subgroup of lantibiotics featured by an indispensable disulfide bridge.lantibiotics are ribosomally-synthesized and posttranslationally modified peptides with potent antimicrobial activities. discovery of novel lantibiotics has been greatly accelerated with the soaring release of genomic information of microorganisms. as a unique class ii lantibiotic, bovicin hj50 is produced by streptococcus bovis hj50 and contains one rare disulfide bridge. by using its precursor bova as a drive sequence, 16 bova-like peptides were revealed in a wide variety of species. from them ...201424821187
clostridium perfringens surgical site infection after osteotomy for knee deformity correction in a non-immunocompromised child.clostridium perfringens myonecrosis following an elective surgical procedure in a previously healthy child is a rare incident.201424828080
development of a flow cytometric bead immunoassay and its assessment as a possible aid to potency evaluation of enterotoxaemia vaccines.enterotoxaemia, an economically important disease of sheep, goats and calves, is caused by systemic effects of the epsilon toxin produced by the anaerobic bacterium clostridium perfringens type d. the only practical means of controlling the occurrence of enterotoxaemia is to immunise animals by vaccination. the vaccine is prepared by deriving a toxoid from the bacterial culture filtrate and the potency of the vaccine is tested with the in vivo mouse neutralisation test (mnt). due to ethical, eco ...201424832497
evaluation of clostridium perfringens as a tracer of sewage contamination in sediments by two enumeration methods.a traditional method of enumerating clostridium perfringens using membrane filtration (mf) as an indicator of fecal contamination was compared to recently developed rapid method using rapid fung double tube (rfdt) in an evaluation to characterize the extent of sewage contamination in sediments of the great lakes. evaluation of these two methods included determining c. perfringens concentrations and recovery efficiencies from sewage, sewage-spiked sediments, and water (surface and bottom) and sed ...201424833022
porcine group a rotaviruses with heterogeneous vp7 and vp4 genotype combinations can be found together with enteric bacteria on belgian swine farms.group a rotaviruses (rva) are an important cause of diarrhea in young piglets, resulting in significant economic losses. however, the role of rva in the etiology of piglet diarrhea on belgian swine farms was previously unreported. in the present study, different techniques, including fast antigen detection tests, virus isolation, rt-pcr and rt-qpcr have been applied for detection of rva in diarrheic (n=28) and asymptomatic (n=6) fecal samples collected on belgian pig farms. rt-qpcr was shown to ...201424837191
investigating an outbreak of clostridium perfringens gastroenteritis in a school using smartphone technology, london, march 2013.on 22 march 2013, 150 of 1,255 students (13–17 years) and staff at a school in london reported gastrointestinal symptoms; onset peaked 8 to 12 hours after a lunch served in the school on 21 march. we performed a retrospective cohort study of all students and staff. we defined cases as school attenders on 20 and 21 march with onset of gastrointestinal symptoms between 20 and 23 march. we tested food, environmental and stool samples of cases for common pathogens and bacterial toxins. we administer ...201424852955
effects of long-term heat stress in an experimental model of avian necrotic enteritis.stressful conditions are predisposing factors for disease development. heat stress is one of the most important stressors in poultry production. the reemergence of some previously controlled diseases [e.g., avian necrotic enteritis (ne)] has been extensively reported. the combination of bacterial infection and certain environmental factors have been reported to trigger the disease. the aim of this study was to analyze the effects of long-term heat stress (35 ± 1°c) on the development of ne in br ...201424879684
differential regulation of microrna transcriptome in chicken lines resistant and susceptible to necrotic enteritis disease.necrotic enteritis (ne) is a re-emerging disease as a result of increased restriction on the use of antibiotics in poultry. however, the molecular mechanisms underlying the pathogenesis of ne are unclear. small rna transcriptome analysis was performed using spleen and intestinal intraepithelial lymphocytes (iel) from 2 inbred chicken lines selected for resistance or susceptibility to marek's disease (md) in an experimentally induced model of avian ne to investigate whether microrna (mirna) contr ...201424879688
haemorrhagic toxin and lethal toxin from clostridium sordellii strain vpi9048: molecular characterization and comparative analysis of substrate specificity of the large clostridial glucosylating toxins.large clostridial glucosylating toxins (lcgts) are produced by toxigenic strains of clostridium difficile, clostridium perfringens, clostridium novyi and clostridium sordellii. while most c. sordellii strains solely produce lethal toxin (tcsl), c. sordellii strain vpi9048 co-produces both hemorrhagic toxin (tcsh) and tcsl. here, the sequences of tcsh-9048 and tcsl-9048 are provided, showing that both toxins retain conserved lcgt features and that tcsl and tcsh are highly related to toxin a (tcda ...201424905543
antimicrobial susceptibility of clinical isolates of anaerobic bacteria in ontario, 2010-2011.the local epidemiology of antimicrobial susceptibility patterns in anaerobic bacteria is important in guiding the empiric treatment of infections. however, susceptibility data are very limited on anaerobic organisms, particularly among non-bacteroides organisms. to determine susceptibility profiles of clinically-significant anaerobic bacteria in ontario canada, anaerobic isolates from sterile sites submitted to public health ontario laboratory (phol) for identification and susceptibility testing ...201424923267
survival of salmonella spp. and fecal indicator bacteria in vietnamese biogas digesters receiving pig slurry.small-scale biogas digesters are widely promoted worldwide as a sustainable technology to manage livestock manure. in vietnam, pig slurry is commonly applied to biogas digesters for production of gas for electricity and cooking with the effluent being used to fertilize field crops, vegetables and fish ponds. slurry may contain a variety of zoonotic pathogens, e.g. salmonella spp., which are able to cause disease in humans either through direct contact with slurry or by fecal contamination of wat ...201424933419
infectious diseases in large-scale cat hoarding investigations.animal hoarders accumulate animals in over-crowded conditions without adequate nutrition, sanitation, and veterinary care. as a result, animals rescued from hoarding frequently have a variety of medical conditions including respiratory infections, gastrointestinal disease, parasitism, malnutrition, and other evidence of neglect. the purpose of this study was to characterize the infectious diseases carried by clinically affected cats and to determine the prevalence of retroviral infections among ...201424934262
clostridium perfringens and clostridium difficile in cooked beef sold in côte d'ivoire and their antimicrobial susceptibility.the aim of this study was to evaluate the prevalence of clostridium difficile and clostridium perfringens in cooked beef sold in the streets in côte d'ivoire and their antimicrobial susceptibility. a total of 395 kidney and flesh samples of cooked beef were collected from vendors at abidjan and subjected to c. difficile and c. perfringens isolation and identification by using biochemical tests, api 20a system and pcr detection. subsequently, the antimicrobial susceptibility test was performed fo ...201424944124
prevention and treatment of clostridium perfringens epsilon toxin intoxication in mice with a neutralizing monoclonal antibody (c4d7) produced in nicotiana benthamiana.epsilon toxin (etx), produced by clostridium perfringens types b and d, is among the most lethal toxins known. etx is a potential bioterrorism threat that was listed as a category b agent by the u.s. centers for disease control until 2012 and it still remains a toxin of interest for several government agencies. we produced a monoclonal antibody (mab) against etx (etx mab c4d7) in nicotiana benthamiana and characterized its preventive and therapeutic efficacy in mice. the etx preparation used was ...201424950050
a claudin 3 and claudin 4-targeted clostridium perfringens protoxin is selectively cytotoxic to psa-producing prostate cancer cells.prostate cancer is the second leading cause of non-cutaneous cancer-related death in males, and effective strategies for treatment of metastatic disease are currently limited. the tight junction proteins, claudin 3 and claudin 4, serve as cell-surface receptors for the pore-forming clostridium perfringens enterotoxin [cpe]. most prostate cancer cells overexpress claudin 3 and claudin 4, and claudins are aberrantly distributed over the plasma membrane, making these cells particularly sensitive to ...201424952257
clostridial pore-forming toxins: powerful virulence factors.pore formation is a common mechanism of action for many bacterial toxins. more than one third of clostridial toxins are pore-forming toxins (pfts) belonging to the β-pft class. they are secreted as soluble monomers rich in β-strands, which recognize a specific receptor on target cells and assemble in oligomers. then, they undergo a conformational change leading to the formation of a β-barrel, which inserts into the lipid bilayer forming functional pore. according to their structure, clostridial ...201424952276
m cell-targeting strategy facilitates mucosal immune response and enhances protection against cvb3-induced viral myocarditis elicited by chitosan-dna vaccine.efficient delivery of antigen to mucosal associated lymphoid tissue is a first and critical step for successful induction of mucosal immunity by vaccines. considering its potential transcytotic capability, m cell has become a more and more attractive target for mucosal vaccines. in this research, we designed an m cell-targeting strategy by which mucosal delivery system chitosan (cs) was endowed with m cell-targeting ability via conjugating with a cpe30 peptide, c terminal 30 amino acids of clost ...201424958702
clostridium perfringens sepsis and liver abscess following laparoscopic cholecystectomy.clostridium perfringens sepsis with intravascular haemolysis is a catastrophic process with a reported mortality of between 90 to 100%. we successfully treated a case of severe clostridial infection with a liver abscess following laparoscopic cholecystectomy, the first to our knowledge. a 59-year-old man presented one week after an uneventful laparoscopic cholecystectomy with jaundice, peritonism, sepsis and acute renal failure. he was found to have a haemolytic anaemia, unconjugated hyperbiliru ...201224960720
application of lactobacillus johnsonii expressing phage endolysin for control of clostridium perfringens.clostridium perfringens is frequently found in food and the environment and produces potent toxins that have a negative impact on both human and animal health and particularly on the poultry industry. lactobacillus johnsonii fi9785, isolated from the chicken gastrointestinal tract, has been demonstrated to exclude cl. perfringens in poultry. we have investigated the interaction of wild-type lact. johnsonii fi9785 or an engineered strain expressing a cell wall-hydrolysing endolysin with cl. perfr ...201424961379
comparative neuropathology of ovine enterotoxemia produced by clostridium perfringens type d wild-type strain cn1020 and its genetically modified derivatives.clostridium perfringens type d causes enterotoxemia in sheep and goats. the disease is mediated by epsilon toxin (etx), which affects the cerebrovascular endothelium, increasing vascular permeability and leading to cerebral edema. in the present study, we compared the distribution and severity of the cerebrovascular changes induced in lambs by c. perfringens type d strain cn1020, its isogenic etx null mutant, and the etx-producing complemented mutant. we also applied histochemical and immunohist ...201524964921
loss of endothelial barrier antigen immunoreactivity as a marker of clostridium perfringens type d epsilon toxin-induced microvascular damage in rat brain.the epsilon toxin elaborated by clostridium perfringens type d in the intestine of domestic livestock is principally responsible for the neurological disease produced after its absorption in excessive quantities into the systemic circulation. the fundamental basis of the cerebral damage induced by epsilon toxin appears to be microvascular injury with ensuing severe, diffuse vasogenic oedema. endothelial barrier antigen (eba), which is normally expressed by virtually all capillaries and venules i ...201524973988
cryptosporidium and giardia in tropical recreational marine waters contaminated with domestic sewage: estimation of bathing-associated disease risks.sewage is a major contributor to pollution problems involving human pathogens in tropical coastal areas. this study investigated the occurrence of intestinal protozoan parasites (giardia and cryptosporidium) in tropical recreational marine waters contaminated with sewage. the potential risks of cryptosporidium and giardia infection from recreational water exposure were estimated from the levels of viable (oo) cysts (dic+, dapi+, pi-) found in near-shore swimming areas using an exponential dose r ...201424975093
clostridium difficile and clostridium perfringens from wild carnivore species in brazil.despite some case reports, the importance of clostridium perfringens and clostridium difficile for wild carnivores remains unclear. thus, the objective of this study was to identify c. perfringens and c. difficile strains in stool samples from wild carnivore species in brazil. a total of 34 stool samples were collected and subjected to c. perfringens and c. difficile isolation. suggestive colonies of c. perfringens were then analyzed for genes encoding the major c. perfringens toxins (alpha, bet ...201424979683
progress and problems in vaccination against necrotic enteritis in broiler chickens.necrotic enteritis in broilers is caused by clostridium perfringens type a strains that produce the netb toxin. necrotic enteritis is one of the gastrointestinal diseases in poultry that has gained worldwide importance during the last decade due to efforts to improve broiler performance. prevention strategies include avoiding predisposing factors, such as coccidiosis, and in-feed supplementation with a variety of feed additives. however, vaccination with modified toxin or other secreted immunoge ...201424980518
public health campaign to promote hand hygiene before meals in a college of veterinary medicine.veterinary students can be exposed to environmental infectious agents in school that may include zoonotic pathogens. encouraging effective hand hygiene can minimize the spread of zoonoses and promote public health and the one health concept among veterinary students. the purpose of this study was to determine if a campaign could improve hand hygiene among veterinary students at extracurricular meetings serving meals. nine kansas state university college of veterinary medicine (ksu-cvm) extracurr ...201424981423
detection of pathogenic clostridia in biogas plant wastes.as the number of biogas plants has grown rapidly in the last decade, the amount of potentially contaminated wastes with pathogenic clostridium spp. has increased as well. this study reports the results from examining 203 biogas plant wastes (bgws). the following clostridium spp. with different frequencies could be isolated via a new enrichment medium (krüne medium) and detected by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (maldi-tof ms): clostridium perfringens ...201524984829
cystathionine β-synthase (cbs) domains confer multiple forms of mg2+-dependent cooperativity to family ii pyrophosphatases.regulated family ii pyrophosphatases (cbs-ppases) contain a nucleotide-binding insert comprising a pair of cystathionine β-synthase (cbs) domains, termed a bateman module. by binding with high affinity to the cbs domains, amp and adp usually inhibit the enzyme, whereas atp activates it. here, we demonstrate that amp, adp, and atp bind in a positively cooperative manner to cbs-ppases from four bacteria: desulfitobacterium hafniense, clostridium novyi, clostridium perfringens, and eggerthella lent ...201424986864
challenging the roles of cd44 and lipolysis stimulated lipoprotein receptor in conveying clostridium perfringens iota toxin cytotoxicity in breast cancer.translational exploration of bacterial toxins has come to the forefront of research given their potential as a chemotherapeutic tool. studies in select tissues have demonstrated that clostridium perfringens iota toxin binds to cd44 and lipolysis stimulated lipoprotein receptor (lsr) cell-surface proteins. we recently demonstrated that lsr expression correlates with estrogen receptor positive breast cancers and that lsr signaling directs aggressive, tumor-initiating cell behaviors. herein, we ide ...201424990559
b-cell epitope of beta toxin of clostridium perfringens genetically conjugated to a carrier protein: expression, purification and characterization of the chimeric protein.beta toxin (btx) is the prime virulence factor for the pathogenesis of clostridium perfringens type c strain, known to cause necrotic enteritis and enterotoxaemia in mammalian species. the existing vaccines targeting btx are formaldehyde inactivated culture filtrates of clostridium. these filtrates raise antigenic load in the host leading to nonspecific and poor responses. the present study aimed to overcome these drawbacks and generate a chimeric protein carrying in silico identified b-cell epi ...201424996028
[a patient with sepsis and a gas-forming liver abscess caused by clostridium perfringens treated with continuous perfusion drainage].a 64-year-old man presented with diarrhea, fever, and disturbance of consciousness; he was subsequently diagnosed with acute renal and hepatic disorder. abdominal computed tomography identified a gas-forming liver abscess, and the patient underwent emergency drainage. however, his condition did not improve, and clostridium perfringens was observed in his blood culture. continuous perfusion drainage was performed by placing an additional drainage tube, which resulted in abscess shrinkage and impr ...201424998733
necrotizing myositis in a neutropenic patient: the use of ultrasound in the diagnosis of atypical presentations.we report a case of fatal necrotizing soft tissue infection (nsti) due to clostridium perfringens (cp) in a neutropenic patient with diabetes mellitus. as in many cases, by the time a diagnosis was made, the condition had rapidly progressed to its late stages, resulting in a fatal outcome. the emergency physician should be aware of nsti as a complication when patients present with pain out of proportion to physical findings and/or signs of soft tissue compromise. negative prognostic factors for ...201425013733
correlation between in vitro cytotoxicity and in vivo lethal activity in mice of epsilon toxin mutants from clostridium perfringens.epsilon toxin (etx) from clostridium perfringens is a pore-forming protein with a lethal effect on livestock, producing severe enterotoxemia characterized by general edema and neurological alterations. site-specific mutations of the toxin are valuable tools to study the cellular and molecular mechanism of the toxin activity. in particular, mutants with paired cysteine substitutions that affect the membrane insertion domain behaved as dominant-negative inhibitors of toxin activity in mdck cells. ...201425013927
cytotoxicity comparison of quercetin and its metabolites from in vitro fermentation of several gut bacteria.part of quercetin is coerced into the colon after ingestion and interacts with the gut microbiota. the interaction between quercetin and gut microbiota will influence human health. the cytotoxicity of quercetin and its metabolites from human gut bacteria in vitro fermentation was investigated in this study. the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide) (mtt) and agar diffusion disc methods were individually applied in vitro to examine their inhibitory effect on three cultured ...201425015032
structure of a c. perfringens enterotoxin mutant in complex with a modified claudin-2 extracellular loop 2.cpe (clostridium perfringens enterotoxin) is the major virulence determinant for c. perfringens type-a food poisoning, the second most common bacterial food-borne illness in the uk and usa. after binding to its receptors, which include particular human claudins, the toxin forms pores in the cell membrane. the mature pore apparently contains a hexamer of cpe, claudin and, possibly, occludin. the combination of high binding specificity with cytotoxicity has resulted in cpe being investigated, with ...201425020226
applicability of universal bacteroidales genetic marker for microbial monitoring of drinking water sources in comparison to conventional indicators.water quality monitoring is essential for the provision of safe drinking water. in this study, we compared a selection of fecal indicators with universal bacteroidales genetic marker to identify fecal pollution of a variety of drinking water sources. a total of 60 samples were collected from water sources. the microbiological parameters included total coliforms, fecal coliforms, escherichia coli and fecal streptococci as the fecal indicator bacteria (fib), clostridium perfringens and h2s bacteri ...201425023746
the effect of experimental fusarium mycotoxicosis on microbiota diversity in porcine ascending colon contents.the objective of the study was to determine the effect of exposure of pigs to the fusarium mycotoxins zearalenone (zen) and deoxynivalenol (don), administered together and separately, on the colon microbiota. an experiment was conducted for 42 days on gilts, randomly assigned to four groups and administered either zen, don, zen+don, or a placebo. the number of aerobic mesophilic bacteria, yeasts, molds, anaerobic clostridium perfringens, fecal streptococci, enterobacteriaceae, escherichia coli, ...201425025709
tailored cyclodextrin pore blocker protects mammalian cells from clostridium difficile binary toxin cdt.some clostridium difficile strains produce, in addition to toxins a and b, the binary toxin clostridium difficile transferase (cdt), which adp-ribosylates actin and may contribute to the hypervirulence of these strains. the separate binding and translocation component cdtb mediates transport of the enzyme component cdta into mammalian target cells. cdtb binds to its receptor on the cell surface, cdta assembles and cdtb/cdta complexes are internalised. in acidic endosomes, cdtb mediates the deliv ...201425029374
[complex surgical management of hemodialysis vascular access infection caused by clostridium perfringens].infection of prosthetic hemodialysis (hd) shunts is one of the most common complications of vascular access in hemodialysis patients. the incidence of anaerobic infection is very rare. in such a case, management of treatment represents a great challenge for the surgeon. we report a complicated case of autologous hemodialysis shunt infected by clostridium perfringens on the right forearm in a polymorbid female patient with chronic renal failure and myelodysplastic syndrome. the patient has underg ...201425047973
effects of dietary additives and early feeding on performance, gut development and immune status of broiler chickens challenged with clostridium perfringens.the effects of dietary additives and holding time on resistance and resilience of broiler chickens to clostridium perfringens challenge were investigated by offering four dietary treatments. these were a negative control (basal), a positive control (zn-bacitracin) and two dietary additives, mannanoligosaccharides (mos), and acidifier. two holding times included (a) immediate access to feed and water post hatch (fed) and (b) access to both feed and water 48 h post hatch (held). chicks fed zn-baci ...201225049595
effect of bacillus subtilis-based direct-fed microbials on immune status in broiler chickens raised on fresh or used litter.type of dietary direct-fed microbials (dfms) or poultry litter could directly influence the composition of gut microbiota. gut microbiota plays an important role in shaping the developing immune system and maintaining the homeostasis of the mature immune system in mammal and chickens. the present study was carried out to investigate the interaction among litter, dfms and immunity in broiler chickens exposed to a field-simulated environment. immune status of broiler chickens was assessed by serum ...201325049746
toxinotyping of clostridium perfringens fecal isolates of reintroduced père david's deer (elaphurus davidianus) in china.clostridium perfringens is an important pathogen causing sudden death syndrome, necrotic enteritis, and gas gangrene in ruminants, especially some deer species. père david's deer (elaphurus davidianus) is one of the world's rare species and is an endangered and protected species in china. some père david's deer in the chinese shishou père david's deer preserve died due to c. perfringens infection. we investigated the toxin types and c. perfringens enterotoxin-positive (cpe(+)) strains of isolate ...201425050802
first isolation of mycoplasma iowae in grey partridge flocks.mycoplasma iowae, an occasional pathogen of turkeys, was isolated for the first time from captive grey partridges (perdix perdix). clinical signs including respiratory and intestinal disorder were seen in birds of all ages but mainly in those kept housed during rearing. mortality rates averaged over 20% during the year. treatment with antibiotics and antiparasitic drugs produced only a transient improvement in condition. the gross pathology findings included poor body growth, lack of development ...201425055642
lumbar discitis caused by clostridium perfringens.we report here a rare case of chronic lumbar discitis caused by clostridium perfringens in an elderly patient that was treated with a combination of β-lactams and clindamycin. molecular analysis performed on the strain revealed an unusual toxin gene pattern.201425056327
cyclophilin-facilitated membrane translocation as pharmacological target to prevent intoxication of mammalian cells by binary clostridial actin adp-ribosylated toxins.clostridium botulinum c2 toxin, clostridium perfringens iota toxin and clostridium difficile cdt belong to the family of binary actin adp-ribosylating toxins and are composed of a binding/translocation component and a separate enzyme component. the enzyme components adp-ribosylate g-actin in the cytosol of target cells resulting in depolymerization of f-actin, cell rounding and cell death. the binding/translocation components bind to their cell receptors and form complexes with the respective en ...201525058685
proportion of illness acquired by foodborne transmission for nine enteric pathogens in australia: an expert elicitation.estimates of the burden of illness acquired from food inform public health policy and prioritize interventions. a key component of such estimates is the proportion of illnesses that are acquired by foodborne transmission. in view of the shortage of requisite data, these proportions are commonly obtained through a process known as expert elicitation. we report findings from an elicitation process used to assess the importance of the foodborne transmission route for nine pathogens in australia, ci ...201425072416
new amino acid germinants for spores of the enterotoxigenic clostridium perfringens type a isolates.clostridium perfringens spore germination plays a critical role in the pathogenesis of c. perfringens-associated food poisoning (fp) and non-food-borne (nfb) gastrointestinal diseases. germination is initiated when bacterial spores sense specific nutrient germinants (such as amino acids) through germinant receptors (grs). in this study, we aimed to identify and characterize amino acid germinants for spores of enterotoxigenic c. perfringens type a. the polar, uncharged amino acids at ph 6.0 effic ...201425084641
screening of lactobacillus strains of domestic goose origin against bacterial poultry pathogens for use as probiotics.lactobacilli are natural inhabitants of human and animal mucous membranes, including the avian gastrointestinal tract. recently, increasing attention has been given to their probiotic, health-promoting capacities, among which their antagonistic potential against pathogens plays a key role. a study was conducted to evaluate probiotic properties of lactobacillus strains isolated from feces or cloacae of domestic geese. among the 104 examined isolates, previously identified to the species level by ...201425104766
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