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diverse mechanisms regulate sporulation sigma factor activity in the firmicutes.sporulation allows bacteria to survive adverse conditions and is essential to the lifecycle of some obligate anaerobes. in bacillus subtilis, the sporulation-specific sigma factors, σ(f), σ(e), σ(g), and σ(k), activate compartment-specific transcriptional programs that drive sporulation through its morphological stages. the regulation of these sigma factors was predicted to be conserved across the firmicutes, since the regulatory proteins controlling their activation are largely conserved. howev ...201525646759
recent insights into clostridium perfringens beta-toxin.clostridium perfringens beta-toxin is a key mediator of necrotizing enterocolitis and enterotoxemia. it is a pore-forming toxin (pft) that exerts cytotoxic effect. experimental investigation using piglet and rabbit intestinal loop models and a mouse infection model apparently showed that beta-toxin is the important pathogenic factor of the organisms. the toxin caused the swelling and disruption of hl-60 cells and formed a functional pore in the lipid raft microdomains of sensitive cells. these f ...201525654787
complementary probes reveal that phosphatidylserine is required for the proper transbilayer distribution of cholesterol.cholesterol is an essential component of metazoan cellular membranes and it helps to maintain the structural integrity and fluidity of the plasma membrane. here, we developed a cholesterol biosensor, termed d4h, based on the fourth domain of clostridium perfringens theta-toxin, which recognizes cholesterol in the cytosolic leaflet of the plasma membrane and organelles. the d4h probe disassociates from the plasma membrane upon cholesterol extraction and after perturbations in cellular cholesterol ...201525663704
tight junctions. structural insight into tight junction disassembly by clostridium perfringens enterotoxin.the c-terminal region of clostridium perfringens enterotoxin (c-cpe) can bind to specific claudins, resulting in the disintegration of tight junctions (tjs) and an increase in the paracellular permeability across epithelial cell sheets. here we present the structure of mammalian claudin-19 in complex with c-cpe at 3.7 å resolution. the structure shows that c-cpe forms extensive hydrophobic and hydrophilic interactions with the two extracellular segments of claudin-19. the claudin-19/c-cpe comple ...201525678664
computing smallest intervention strategies for multiple metabolic networks in a boolean model.this article considers the problem whereby, given two metabolic networks n1 and n2, a set of source compounds, and a set of target compounds, we must find the minimum set of reactions whose removal (knockout) ensures that the target compounds are not producible in n1 but are producible in n2. similar studies exist for the problem of finding the minimum knockout with the smallest side effect for a single network. however, if technologies of external perturbations are advanced in the near future, ...201525684199
clostridium perfringens bacteremia in an 85-year-old diabetic man.emphysematous cholecystitis is an uncommon and dangerous complication of acute cholecystitis. common risk factors for this disease include male gender, old age, presence of diabetes mellitus and cholelithiasis. the disease is best treated with emergent surgery and parenteral antibiotics. we present the case of an 85-year-old nursing home resident who presented to our institution with a 3-day history of gradually worsening abdominal discomfort.201725685130
solution structure and dna binding of the catalytic domain of the large serine resolvase tnpx.the transfer of antibiotic resistance between bacteria is mediated by mobile genetic elements such as plasmids and transposons. tnpx is a member of the large serine recombinase subgroup of site-specific recombinases and is responsible for the excision and insertion of mobile genetic elements that encode chloramphenicol resistance in the pathogens clostridium perfringens and clostridium difficile. tnpx consists of three structural domains: domain i contains the catalytic site, whereas domains ii ...201525720550
microbial pathogens in wastewater treatment plants (wwtp) in hamburg.microbial pathogens are among the major health problems associated with water and wastewater. classical indicators of fecal contamination include total coliforms, escherichia coli, and clostridium perfringens. these fecal indicators were monitored in order to obtain information regarding their evolution during wastewater treatment processes. helminth eggs survive for a long duration in the environment and have a high potential for waterborne transmission, making them reliable contaminant indicat ...201525734765
characteristics of metroxylon sagu resistant starch type iii as prebiotic substance.resistant starch type iii (rs3 ) was produced from sago (metroxylon sagu) and evaluated for its characteristics as a prebiotic. two rs3 samples designated sago rs and hcl-sago rs contained 35.71% and 68.30% rs, respectively, were subjected to hydrolyses by gastric juice and digestive enzymes and to absorption. both sago rs and hcl-sago rs were resistant to 180 min hydrolysis by gastric acidity at ph 1 to 4 with less than 0.85% hydrolyzed. both samples were also resistant toward hydrolysis by gas ...201525739421
characterization of deep sea fish gut bacteria with antagonistic potential, from centroscyllium fabricii (deep sea shark).the bacterial isolates from centroscyllium fabricii (deep sea shark) gut were screened for antagonistic activity by cross-streak method and disc diffusion assay. this study focuses on strain btss-3, which showed antimicrobial activity against pathogenic bacteria including salmonella typhimurium, proteus vulgaris, clostridium perfringens, staphylococcus aureus, bacillus cereus, bacillus circulans, bacillus macerans and bacillus pumilus. btss3 was subjected to phenotypic characterization using bio ...201525740801
loop-mediated isothermal amplification (lamp) assays for the species-specific detection of eimeria that infect chickens.eimeria species parasites, protozoa which cause the enteric disease coccidiosis, pose a serious threat to the production and welfare of chickens. in the absence of effective control clinical coccidiosis can be devastating. resistance to the chemoprophylactics frequently used to control eimeria is common and sub-clinical infection is widespread, influencing feed conversion ratios and susceptibility to other pathogens such as clostridium perfringens. despite the availability of polymerase chain re ...201525741643
[a case of freeze-dried gas gangrene antitoxin for the treatment of clostridium perfringens sepsis].a 66-year-old man was admitted to our hospital with high fever. we diagnosed a gas-containing liver abscess and performed percutaneous abscess drainage. however, 15 hours after admission, he developed massive intravascular hemolysis and acidosis. sepsis due to clostridium perfringens was suspected and we treated the patient intensively with multidisciplinary approaches, including antibiotics, mechanical ventilation, and renal replacement therapy. furthermore, we administered freeze-dried gas gan ...201525748160
an allosteric model for control of pore opening by substrate binding in the eutl microcompartment shell protein.the ethanolamine utilization (eut) microcompartment is a protein-based metabolic organelle that is strongly associated with pathogenesis in bacteria that inhabit the human gut. the exterior shell of this elaborate protein complex is composed from a few thousand copies of bmc-domain shell proteins, which form a semi-permeable diffusion barrier that provides the interior enzymes with substrates and cofactors while simultaneously retaining metabolic intermediates. the ability of this protein shell ...201525752492
epidemiological and pathobiological profiles of clostridium perfringens infections: review of consecutive series of 33 cases over a 13-year period.although clostridium perfringens (c. perfringens) is well known as the causative agent of several forms of enteric disease, precise epidemiological and pathobiological aspects are still unknown.201525755747
use of yeast cell wall extract as a tool to reduce the impact of necrotic enteritis in broilers.the use of a yeast cell wall extract derived from saccharomyces cerevisiae (actigen(®)) has been proposed as an alternative to in-feed antibiotics. this experiment was conducted to investigate the efficacy of yeast cell extract as an alternative to zinc bacitracin or salinomycin using a necrotic enteritis challenge model. a feeding study was conducted using 480-day-old male ross 308 chicks assigned to 48 floor pens. a 2 × 4 factorial arrangement of treatments was employed. the factors were: chal ...201525762162
health risk assessment related to waterborne pathogens from the river to the tap.a two-year monitoring program of cryptosporidium parvum oocysts, giardia duodenalis cysts, escherichia coli, clostridium perfringens spores and adenovirus was conducted in three large rivers in france used for recreational activities and as a resource for drinking water production. fifty-liter river water and one thousand-liter tap water samples were concentrated using hollow-fiber ultrafiltration and analyzed by molecular biology or laser-scanning cytometry. in order to evaluate watershed land ...201525764059
clostridial co-infection episodes in commercial laying hens.the present report describes two outbreaks of serious enteritis in commercial laying hens where clostridium perfringens and clostridium colinum were simultaneously detected. at the age of 44 and 31 weeks, two laying hen flocks showed an increase of the mortality rate and a worsening of productive performance. post-mortem examination revealed intestinal necrotic-haemorrhagic ulcerations and hepatic focal necrosis. the bacteriological examination yielded the isolation of c. colinum and c. perfring ...201525769045
animal models to study the pathogenesis of human and animal clostridium perfringens infections.the most common animal models used to study clostridium perfringens infections in humans and animals are reviewed here. the classical c. perfringens-mediated histotoxic disease of humans is clostridial myonecrosis or gas gangrene and the use of a mouse myonecrosis model coupled with genetic studies has contributed greatly to our understanding of disease pathogenesis. similarly, the use of a chicken model has enhanced our understanding of type a-mediated necrotic enteritis in poultry and has led ...201525770894
structure-function analysis of peptide signaling in the clostridium perfringens agr-like quorum sensing system.the accessory growth regulator (agr)-like quorum sensing (qs) system of clostridium perfringens controls the production of many toxins, including beta toxin (cpb). we previously showed (j. e. vidal, m. ma, j. saputo, j. garcia, f. a. uzal, and b. a. mcclane, mol microbiol 83:179-194, 2012, http://dx.doi.org/10.1111/j.1365-2958.2011.07925.x) that an 8-amino-acid, agrd-derived peptide named 8-r upregulates cpb production by this qs system. the current study synthesized a series of small signaling ...201525777675
phytase modulates ileal microbiota and enhances growth performance of the broiler chickens.phytase is well studied and explored, however, little is known about its effects on the microbial ecology of the gastrointestinal tract. in total, 400 one-day-old female ross 308 chicks were randomly distributed to four experimental groups. the dietary treatments were arranged as a 2 × 2 complete factorial design, with the factors being adequate (pc) or insufficient calcium (ca) and digestible phosphor (dp)(nc) and with or without 5000 phytase units (ftu)/kg of escherichia coli 6-phytase. the ga ...201525781608
improvement of immunity and disease resistance in the nile tilapia, oreochromis niloticus, by dietary supplementation with bacillus amyloliquefaciens.probiotics can be used as immunostimulants in aquaculture. the aim of this study was to evaluate the immune responses of nile tilapia oreochromis niloticus following feeding with bacillus amyloliquefaciens spores at concentrations of 1 × 10(6) (g3) and 1 × 10(4) (g2) colony-forming units per gram (cfu/g) of feed compared with a basal diet with no probiotics (g1). a total of 180 fingerlings (27.7 ± 0.22 g) were divided into three groups (g1-g3 of 20 fish per group) in triplicate. innate immunitie ...201525783002
hygienisation and nutrient conservation of sewage sludge or cattle manure by lactic acid fermentation.manure from animal farms and sewage sludge contain pathogens and opportunistic organisms in various concentrations depending on the health of the herds and human sources. other than for the presence of pathogens, these waste substances are excellent nutrient sources and constitute a preferred organic fertilizer. however, because of the pathogens, the risks of infection of animals or humans increase with the indiscriminate use of manure, especially liquid manure or sludge, for agriculture. this p ...201525786255
the inhibitory effects of sorbate and benzoate against clostridium perfringens type a isolates.this study evaluated the inhibitory effects of sorbate and benzoate against clostridium perfringens type a food poisoning (fp) and non-food-borne (nfb) disease isolates. no significant inhibition of germination of spores of both fp and nfb isolates was observed in rich medium (ph 7.0) supplemented with permissive level of sodium sorbate (0.3% ≈ 0.13 mm undissociated sorbic acid) or sodium benzoate (0.1% ≈ 0.01 mm undissociated benzoic acid) used in foods. however, these levels of sorbate and ben ...201525790996
clostridium perfringens type e virulence traits involved in gut colonization.clostridium perfringens type e disease in ruminants has been characterized by hemorrhagic enteritis or sudden death. although type e isolates are defined by the production of alpha and iota toxin, little is known about the pathogenesis of c. perfringens type e infections. thus far, the role of iota toxin as a virulence factor is unknown. in this report, iota toxin showed positive effects on adherence and colonization of c. perfringens type e while having negative effect on the adherence of type ...201525799452
effect of lactobacillus plantarum strain k21 on high-fat diet-fed obese mice.recent studies have demonstrated beneficial effects of specific probiotics on alleviating obesity-related disorders. here we aimed to identify probiotics with potential antiobesity activity among 88 lactic acid bacterial strains via in vitro screening assays, and a lactobacillus plantarum strain k21 was found to harbor abilities required for hydrolyzing bile salt, reducing cholesterol, and inhibiting the accumulation of lipid in 3t3-l1 preadipocytes. furthermore, effects of k21 on diet-induced o ...201525802537
first detection of macrorhabdus ornithogaster in wild eurasian siskins (carduelis spinus) in germany. a case study.the colonization of the gastric ascomycetous yeast macrorhabdus (m.) ornithogaster could be associated with a chronic wasting disease in several bird species in captivity. the prevalence and clinical relevance of m. ornithogaster in wild birds is unknown in detail.201525804259
multi-organ failure secondary to a clostridium perfringens gaseous liver abscess following a self-limited episode of acute gastroenteritis.clostridium perfringens is an unusual pathogen responsible for the development of a gas-forming pyogenic liver abscess. progression to septicemia with this infection has amplified case fatality rates.201525807198
characterization of clostridium perfringens tpel toxin gene carriage, production, cytotoxic contributions, and trypsin sensitivity.large clostridial toxins (lcts) are produced by at least four pathogenic clostridial species, and several lcts are proven pivotal virulence factors for both human and veterinary diseases. tpel is a recently identified lct produced by clostridium perfringens that has received relatively limited study. in response, the current study surveyed carriage of the tpel gene among different c. perfringens strains, detecting this toxin gene in some type a, b, and c strains but not in any type d or e strain ...201525824828
the cpal quorum sensing system regulates production of hemolysins cpa and pfo to build clostridium perfringens biofilms.clostridium perfringens strains produce severe diseases, including myonecrosis and enteritis necroticans, in humans and animals. diseases are mediated by the production of potent toxins that often damage the site of infection, e.g., skin epithelium during myonecrosis. in planktonic cultures, the regulation of important toxins, such as cpa, cpb, and pfo, is controlled by the c. perfringens agr-like (cpal) quorum sensing (qs) system. strains also encode a functional luxs/ai-2 system. although c. p ...201525824838
focal bacterial meningitis following ascending bite wound infection leading to paraparesis in a captive california sea lion (zalophus californianus).magnetic resonance imaging was performed on a 15-yr-old captive female california sea lion (zalophus californianus) with a 2-wk history of progressive paraparesis and a 9-mo history of exudative skin lesion on the left thoracic wall. magnetic resonance images showed a well-defined muscle infiltrating lesion ventrolateral to the seventh cervical to the third thoracic vertebra on the left side, which extended through the left intervertebral foramina c7 to t3 into the vertebral canal, causing spina ...201525831587
clostridium perfringens enterotoxin (cpe) and cpe-binding domain (c-cpe) for the detection and treatment of gynecologic cancers.clostridium perfringens enterotoxin (cpe) is a three-domain polypeptide, which binds to claudin-3 and claudin-4 with high affinity. because these receptors are highly differentially expressed in many human tumors, claudin-3 and claudin-4 may provide an efficient molecular tool to specifically identify and target biologically aggressive human cancer cells for cpe-specific binding and cytolysis. in this review we will discuss these surface proteins as targets for the detection and treatment of che ...201525835384
expert elicitation as a means to attribute 28 enteric pathogens to foodborne, waterborne, animal contact, and person-to-person transmission routes in canada.enteric illness contributes to a significant burden of illness in canada and globally. understanding its sources is a critical step in identifying and preventing health risks. expert elicitation is a powerful tool, used previously, to obtain information about enteric illness source attribution where information is difficult or expensive to obtain. thirty-one experts estimated transmission of 28 pathogens via major transmission routes (foodborne, waterborne, animal contact, person-to-person, and ...201525835810
effect of oxygen stress on growth and survival of clostridium perfringens, campylobacter jejuni, and listeria monocytogenes under different storage conditions.this study investigated the growth and survival of three foodborne pathogens (clostridium perfringens, campylobacter jejuni, and listeria monocytogenes) in beef (7% fat) and nutrient broth under different oxygen levels. samples were tested under anoxic (<0.5%), microoxic (6 to 8%), and oxic (20%) conditions during storage at 7 °c for 14 days and at 22 °c for 5 days. two initial inoculum concentrations were used (1 and 2 log cfu per g of beef or per ml of broth). the results show that c. perfring ...201525836393
microbiological diversity and prevalence of spoilage and pathogenic bacteria in commercial fermented alcoholic beverages (beer, fruit wine, refined rice wine, and yakju).the present study examined 469 commercially available fermented alcoholic beverages (fabs), including beer (draft, microbrewed, and pasteurized), fruit wine (grape and others), refined rice wine, and yakju (raw and pasteurized). samples were screened for escherichia coli and eight foodborne pathogens (bacillus cereus, campylobacter jejuni, clostridium perfringens, escherichia coli o157:h7, listeria monocytogenes, salmonella spp., staphylococcus aureus, and yersinia enterocolitica), and the aerob ...201525836410
congener specific polychlorinated biphenyl metabolism by human intestinal microbe clostridium species: comparison with human liver cell line-hepg2.polychlorobiphenyls (pcbs), which adversely affect human fetal and infant development, are endocrine disrupter and cause neurological disorders. they may also be carcinogenic. it is not known whether these effects are due to whole pcbs or to its metabolites, produced by the human gastrointestinal system primarily the liver and/or by intestinal microbes such as clostridium sp. the available data show that clostridium perfringens, the most prominent species of clostridium occurs in the human gut. ...200625838614
characterization of clostridium perfringens isolates obtained from 2010 to 2012 from chickens with necrotic enteritis in korea.clostridium perfringens produces diverse virulent toxins that cause necrotic enteritis in poultry, resulting in a great negative impact on the poultry industry. to study the characteristics of c. perfringens in chickens, we isolated 88 strains from chickens (1 strain per flock) with necrotic enteritis. the isolated bacterial strains were screened for toxin type and antimicrobial susceptibility. necropsy of 17 chickens that died from necrotic enteritis revealed that their intestines were dilated ...201525840962
bacterial enteritis in ostrich (struthio camelus) chicks in the western cape province, south africa.ostrich (struthio camelus) chicks less than 3 mo age are observed to experience a high mortality rate that is often associated with enteritis. this study was undertaken to investigate the infectious bacteria implicated in ostrich chick enteritis. postmortems were performed on 122 ostrich chicks aged from 1 d to 3 mo and intestinal samples were subjected to bacterial culture. bacterial isolates were typed by pcr and serotyping. escherichia coli (e. coli; 49%) was the most frequently isolated from ...201525840967
claudin-4 spect imaging allows detection of aplastic lesions in a mouse model of breast cancer.the expression of claudin-4, a protein involved in tight junction complexes, is widely dysregulated in epithelial malignancies. claudin-4 is overexpressed in several premalignant precursor lesions, including those of cancers of the breast, pancreas, and prostate, and is associated with poor survival. a noncytotoxic c-terminal fragment of clostridium perfringens enterotoxin (ccpe) is a natural ligand for claudin-4. here, we demonstrate whole-body quantitative spect imaging of preneoplastic breast ...201525840973
rye affects bacterial translocation, intestinal viscosity, microbiota composition and bone mineralization in turkey poults.previously, we have reported that rye significantly increased both viscosity and clostridium perfringens proliferation when compared with corn in an in vitro digestive model. two independent trials were conducted to evaluate the effect of rye as a source of energy on bacterial translocation, intestinal viscosity, gut microbiota composition, and bone mineralization, when compared with corn in turkey poults. in each experiment, day-of-hatch, turkey poults were randomly assigned to either a corn or ...201525849537
isolation and characterization of a new [fefe]-hydrogenase from clostridium perfringens.this paper reports the first characterization of an [fefe]-hydrogenase from a clostridium perfringens strain previously isolated in our laboratory from a pilot-scale bio-hydrogen plant that efficiently produces h2 from waste biomasses. on the basis of sequence analysis, the enzyme is a monomer formed by four domains hosting various iron-sulfur centres involved in electron transfer and the catalytic center h-cluster. after recombinant expression in escherichia coli, the purified protein catalyzes ...201625851509
neuroinflammation induced by intracerebroventricular injection of microbial neuraminidase.in the present paper, we describe the facts that took place in the rat brain after a single injection of the enzyme neuraminidase from clostridium perfringens into the right lateral ventricle. after injection, it diffused through the cerebrospinal fluid of the ipsilateral ventricle and the third ventricle, and about 400 μm into the periventricular brain parenchyma. the expression of icam1 in the endothelial cells of the periventricular vessels, iba1 in microglia, and gfap in astrocytes notably i ...201525853134
a novel pore-forming toxin in type a clostridium perfringens is associated with both fatal canine hemorrhagic gastroenteritis and fatal foal necrotizing enterocolitis.a role for type a clostridium perfringens in acute hemorrhagic and necrotizing gastroenteritis in dogs and in necrotizing enterocolitis of neonatal foals has long been suspected but incompletely characterized. the supernatants of an isolate made from a dog and from a foal that died from these diseases were both found to be highly cytotoxic for an equine ovarian (eo) cell line. partial genome sequencing of the canine isolate revealed three novel putative toxin genes encoding proteins related to t ...201525853427
antibiotic-resistant bacteria: prevalence in food and inactivation by food-compatible compounds and plant extracts.foodborne antibiotic-resistant pathogenic bacteria such as campylobacter jejuni, bacillus cereus, clostridium perfringens, escherichia coli, salmonella enterica, staphylococcus aureus, vibrio cholerae, and vibrio parahemolyticus can adversely affect animal and human health, but a better understanding of the factors involved in their pathogenesis is needed. to help meet this need, this overview surveys and interprets much of our current knowledge of antibiotic (multidrug)-resistant bacteria in th ...201525856120
attack rate in food poisoning: order in chaos.the frequency distributions of the numbers of patients and eaters were near lognormal for almost all food poisonings. the medians and ranges remained almost unchanged over time, although the annual number of incidents varied. however, the numbers of patients and eaters were not correlated for many of the food poisonings. a regular pattern relating the numbers of patients and eaters emerged for food poisonings exhibited strong seasonal incidence. for example, food poisonings caused by norovirus o ...201525866112
specific binding of clostridium perfringens enterotoxin fragment to claudin-b and modulation of zebrafish epidermal barrier.claudins (cldn) are the major components of tight junctions (tjs) sealing the paracellular cleft in tissue barriers of various organs. zebrafish cldnb, the homolog of mammalian cldn4, is expressed at epithelial cell-cell contacts and is important for regulating epidermal permeability. the bacterial toxin clostridium perfringens enterotoxin (cpe) has been shown to bind to a subset of mammalian cldns. in this study, we used the cldn-binding c-terminal domain of cpe (194-319 amino acids, ccpe 194-3 ...201525869230
characterization of the spore-forming bacillus cereus sensu lato group and clostridium perfringens bacteria isolated from the australian dairy farm environment.the bacillus cereus sensu lato group and clostridium perfringens are spore-forming bacteria often associated with food spoilage and which can cause emetic and diarrheal syndromes in humans and ruminants. this study characterised the phenotypes and genotypes of 50 bacillus cereus s. l. isolates and 26 clostridium perfringens isolates from dairy farms environments in victoria, australia.201525881096
non-classical azoreductase secretion in clostridium perfringens in response to sulfonated azo dye exposure.clostridium perfringens, a strictly anaerobic microorganism and inhabitant of the human intestine, has been shown to produce an azoreductase enzyme (azoc), an nadh-dependent flavin oxidoreductase. this enzyme reduces azo dyes into aromatic amines, which can be carcinogenic. a significant amount of work has been completed on the activity of azoc. despite this, much is still unknown, including whether azoreduction of these dyes occurs intracellularly or extracellulary. a physiological study of c.  ...201525881497
vertebral osteomyelitis and epidural abscess caused by gas gangrene presenting with complete paraplegia: a case report.gas gangrene is most often caused by clostridium perfringens infection. gas gangrene is a medical emergency that develops suddenly. the mortality rate is higher with trunk involvement than with involvement of the extremities, which carries a better prognosis. with respect to vertebral involvement, there are few reports in the literature. the purpose of this paper is to report a very rare case of vertebral osteomyelitis caused by gas gangrene.201525888739
development of intestinal microflora and occurrence of diarrhoea in sucking foals: effects of bacillus cereus var. toyoi supplementation.almost all foals develop transient diarrhoea within the first weeks of life. studies indicated different viral, bacterial, and parasitic causes, such as rotavirus, clostridium perfringens, escherichia coli, and cryptosporidium are discussed. but little is known about the development of intestinal microflora in foals. the present study investigated whether the supplementation with bacillus cereus var. toyoi would modify the developing intestinal microflora and consequently reduce diarrhoea in foa ...201525889817
clostridium perfringens infection after transarterial chemoembolization for large hepatocellular carcinoma.we report an unusual case of clostridium perfringens liver abscess formation after transcatheter arterial chemoembolization (tace) for large hepatocellular carcinoma. severe deterioration in liver and renal function accompanied with hemocytolysis was found on the 2(nd) day after tace. blood culture found clostridium perfringens and abdominal computed tomography revealed a gas-containing abscess in the liver. following antibiotics administration and support care, the infection was controlled and ...201525892893
bioassay-guided chromatographic isolation and identification of antibacterial compounds from artemisia annua l. that inhibit clostridium perfringens growth.clostridium perfringens is the causative agent of necrotic enteritis leading to significant losses in the poultry industry. dichloromethane and n-hexane extracts of aerial parts of artemisia annua (asteraceae) exhibited activity against c. perfringens with minimum inhibitory concentrations (mic) of 185 and 270 μg/ml, respectively. bioassay-guided fractionation of the extracts gave several active fractions (mic between 75 and 600 μg/ml). investigations of the most active fractions resulted in the ...201625902977
effect of a probiotic on prevention of diarrhea and clostridium difficile and clostridium perfringens shedding in foals.up to 60% of foals develop diarrhea within 6 months after birth. preventive measures are limited but potentially probiotics could be used.201725903509
clostridium perfringens alpha-toxin induces gm1a clustering and trka phosphorylation in the host cell membrane.clostridium perfringens alpha-toxin elicits various immune responses such as the release of cytokines, chemokines, and superoxide via the gm1a/trka complex. alpha-toxin possesses phospholipase c (plc) hydrolytic activity that contributes to signal transduction in the pathogenesis of gas gangrene. little is known about the relationship between lipid metabolism and trka activation by alpha-toxin. using live-cell fluorescence microscopy, we monitored transbilayer movement of diacylglycerol (dag) wi ...201525910247
use of power ultrasound to enhance the thermal inactivation of clostridium perfringens spores in beef slurry.clostridium perfringens is a pathogen of concern in pasteurised foods. the main objective of this study was to use power ultrasound to enhance the thermal inactivation of c. perfringens spores in beef slurry. the effect of simultaneous ultrasound and heat (ts, thermosonication) on the spore inactivation in beef slurry was first investigated. at 75 °c, a 60 min ts process (24 khz, 0.33 w/g) resulted in a less than 1.5 log reduction for both c. perfringens nzrm 898 and nzrm 2621 spores. then, the ...201525912313
in vitro cytotoxicity induced by clostridium perfringens isolate carrying a chromosomal cpe gene is exclusively dependent on sporulation and enterotoxin production.clostridium perfringens type a is a common source of food poisoning (fp) and non-food-borne (nfb) gastrointestinal diseases in humans. in the intestinal tract, the vegetative cells sporulate and produce a major pathogenic factor, c. perfringens enterotoxin (cpe). most type a fp isolates carry a chromosomal cpe gene, whereas nfb type a isolates typically carry a plasmid-encoded cpe. in vitro, the purified cpe protein binds to a receptor and forms pores, exerting a cytotoxic activity in epithelial ...201525912832
amino acid residue y196e substitution and c-terminal peptide synergistically alleviate the toxicity of clostridium perfringens epsilon toxin.epsilon toxin (etx) is produced by clostridium perfringens type b and d strains, and is the causative agent of a lethal enterotoxemia in livestock animals and possibly in humans. however, many details of etx structure and activity are not known. therefore, it is important to clarify the relationship between etx structure and activity. to explore the effect and mechanism of etx amino acid residue y196e substitution and c-terminal peptide on toxicity, four recombinant proteins, retx (without 13 n- ...201525912943
the role of biological processes in reducing both odor impact and pathogen content during mesophilic anaerobic digestion.mesophilic anaerobic digestion (mad) produces renewable energy, but it also plays a role in reducing the impact of digestates, both by reducing odor and pathogen content. ten full-scale biogas plants characterized by different plant designs (e.g. single digesters, parallel or serial digesters), plant powers (ranging from 180 to 999 kwe), hydraulic retention time (hrt) (ranging between 20 to 70 days) and feed mixes were monitored and odors and pathogens were observed in both ingestates and digest ...201525925189
plasmid partitioning systems of conjugative plasmids from clostridium perfringens.many pathogenic strains of clostridium perfringens carry several highly similar toxin or antibiotic resistance plasmids that have 35 to 40 kb of very closely related syntenous sequences, including regions that carry the genes encoding conjugative transfer, plasmid replication and plasmid maintenance functions. key questions are how are these closely related plasmids stably maintained in the same cell and what is the basis for plasmid incompatibility in c. perfringens. comparative analysis of the ...201525929175
recovery of staphylococcus aureus in gray mugil cephalus roe (bottarga): investigation by an integrated cultural/molecular approach.in the mediterranean area, salted and dried roe from the gray mugil cephalus "bottarga" represent a speciality food with great commercial value. bottarga is currently produced by a traditional handmade process and, the risk of human bacterial contamination during its manufacturing is still unknown; in this perspective the foodborne pathogen staphylococcus aureus could potentially contaminate this product due to poor sanitation or bad handling during processing. the aim of this work is: to evalua ...201525944563
clostridium perfringens tpel induces formation of stress fibers via activation of rhoa-rock signaling pathway.clostridium perfringens tpel belongs to a family of large clostridial glucosylating cytotoxins. tpel modifies rac1 and ras subfamily proteins. herein we report tpel-induced formation of stress fibers via rhoa-rho kinase (rock) signaling. a recombinant protein (tpel1-525) derived from the tpel n-terminal catalytic domain in the presence of streptolysin o (slo) induced the formation of actin stress fibers in madin-darby canine kidney (mdck) cells in a dose-dependent manner. the rhoa/rock pathway i ...201525947919
impact of clean-label antimicrobials and nitrite derived from natural sources on the outgrowth of clostridium perfringens during cooling of deli-style turkey breast.organic acids and sodium nitrite have long been shown to provide antimicrobial activity during chilling of cured meat products. however, neither purified organic acids nor nano2 is permitted in products labeled natural and both are generally avoided in clean-label formulations; efficacy of their replacement is not well understood. natural and clean-label antimicrobial alternatives were evaluated in both uncured and in alternative cured (a process that uses natural sources of nitrite) deli-style ...201525951389
genotyping of clostridium perfringens isolated from healthy and diseased ostriches (struthio camelus).clostridium perfringens is more prevalent type of clostridia genus isolated from the intestinal tract of ostrich (struthio camelus). necrotic enteritis (ne) is a potentially fatal gastrointestinal (gi) disease of poultry and other avian species, which produces marked destruction of intestinal lining in digestive tract caused by c. perfringens. pathogenicity and lesions are correlated with the toxins produced, thus toxin typing of the bacterium has diagnostic and epidemiological significance. the ...201425954489
assembly and function of claudins: structure-function relationships based on homology models and crystal structures.the tetra-span transmembrane proteins of the claudin family are critical components of formation and function of tight junctions (tj). homo- and heterophilic side-by-side (cis) and intercellular head-to-head (trans) interactions of 27 claudin-subtypes regulate tissue-specifically the paracellular permeability and/or tightness between epithelial or endothelial cells. this review highlights the functional impact that has been identified for particular claudin residues by relating them to structura ...201525957516
enforcement of science-using a clostridium perfringens outbreak investigation to take legal action.we report an outbreak of clostridium perfringens in a care home in north east england.201625972386
genome-wide transcriptional profiling of clostridium perfringens sm101 during sporulation extends the core of putative sporulation genes and genes determining spore properties and germination characteristics.the formation of bacterial spores is a highly regulated process and the ultimate properties of the spores are determined during sporulation and subsequent maturation. a wide variety of genes that are expressed during sporulation determine spore properties such as resistance to heat and other adverse environmental conditions, dormancy and germination responses. in this study we characterized the sporulation phases of c. perfringens enterotoxic strain sm101 based on morphological characteristics, ...201525978838
clostridium sordellii genome analysis reveals plasmid localized toxin genes encoded within pathogenicity loci.clostridium sordellii can cause severe infections in animals and humans, the latter associated with trauma, toxic shock and often-fatal gynaecological infections. strains can produce two large clostridial cytotoxins (lccs), tcsl and tcsh, related to those produced by clostridium difficile, clostridium novyi and clostridium perfringens, but the genetic basis of toxin production remains uncharacterised.201525981746
fate and transport of enteric microbes from septic systems in a coastal watershed.onsite wastewater treatment systems (owts) are commonly used in coastal areas to treat household wastewater. these systems represent potential sources of fecal pollution of groundwater and nearby surface water. owts are expected to reduce microbial concentrations in wastewater; however, system and environmental factors can affect treatment efficiency and impacts on ground and surface water. in the study of owts described in this article, the authors sampled septic tanks and groundwater at two ho ...201525985535
impact of mutating the key residues of a bifunctional 5,10-methylenetetrahydrofolate dehydrogenase-cyclohydrolase from escherichia coli on its activities.methylenetetrahydrofolate dehydrogenase-cyclohydrolase (fold) catalyzes interconversion of 5,10-methylene-tetrahydrofolate and 10-formyl-tetrahydrofolate in the one-carbon metabolic pathway. in some organisms, the essential requirement of 10-formyl-tetrahydrofolate may also be fulfilled by formyltetrahydrofolate synthetase (fhs). recently, we developed an escherichia coli strain in which the fold gene was deleted in the presence of clostridium perfringens fhs (e. coli δfold/p-fhs) and used it to ...201525988590
the myelin and lymphocyte protein mal is required for binding and activity of clostridium perfringens ε-toxin.clostridium perfringens ε-toxin (etx) is a potent pore-forming toxin responsible for a central nervous system (cns) disease in ruminant animals with characteristics of blood-brain barrier (bbb) dysfunction and white matter injury. etx has been proposed as a potential causative agent for multiple sclerosis (ms), a human disease that begins with bbb breakdown and injury to myelin forming cells of the cns. the receptor for etx is unknown. here we show that both binding of etx to mammalian cells and ...201525993478
characterization, validation and application of a dna microarray for the detection of mandatory and other waterborne pathogens.culture methods for the detection of indicator bacteria are currently used for detection of waterborne bacteria. the need for an increased range of analyzed bacteria coupled with the obtainment of rapid and early results justify the development of a dna microarray for the identification of waterborne pathogens. this dna microarray has 16 implanted probes with a median size of 147 bases, targeting 12 different parameters, including all mandatory indicator microorganisms, such as escherichia coli, ...201525998249
location and stoichiometry of the protease cspb and the cortex-lytic enzyme slec in clostridium perfringens spores.the protease cspb and the cortex-lytic enzyme slec are essential for peptoglycan cortex hydrolysis during germination of spores of the clostridium perfringens food poisoning isolate sm101. in this study, western blot analyses were used to demonstrate that cspb and slec are present exclusively in the c. perfringens sm101 spore coat layer fraction and absent in the lysate from decoated spores and from the purified inner spore membrane. these results indicate why decoating treatments greatly reduce ...201525998819
claudin-related intestinal diseases.with up to 200 m(2) the human intestine is the organ with the largest absorptive surface of the body. it is lined by a single layer of epithelial cells that separates the host from the environment. the intestinal epithelium provides both, selective absorption of nutrients, ions, and water but also a highly effective barrier function which includes the first line of defense against environmental antigens. the paracellular part of this barrier function is provided by tight junction (tj) proteins, ...201525999319
automated sampling procedures supported by high persistence of bacterial fecal indicators and bacteroidetes genetic microbial source tracking markers in municipal wastewater during short-term storage at 5°c.because of high diurnal water quality fluctuations in raw municipal wastewater, the use of proportional autosampling over a period of 24 h at municipal wastewater treatment plants (wwtps) to evaluate carbon, nitrogen, and phosphorus removal has become a standard in many countries. microbial removal or load estimation at municipal wwtps, however, is still based on manually recovered grab samples. the goal of this study was to establish basic knowledge regarding the persistence of standard bacteri ...201526002900
perfringolysin o: the underrated clostridium perfringens toxin?the anaerobic bacterium clostridium perfringens expresses multiple toxins that promote disease development in both humans and animals. one such toxin is perfringolysin o (pfo, classically referred to as θ toxin), a pore-forming cholesterol-dependent cytolysin (cdc). pfo is secreted as a water-soluble monomer that recognizes and binds membranes via cholesterol. membrane-bound monomers undergo structural changes that culminate in the formation of an oligomerized prepore complex on the membrane sur ...201526008232
c-terminal clostridium perfringens enterotoxin-mediated antigen delivery for nasal pneumococcal vaccine.efficient vaccine delivery to mucosal tissues including mucosa-associated lymphoid tissues is essential for the development of mucosal vaccine. we previously reported that claudin-4 was highly expressed on the epithelium of nasopharynx-associated lymphoid tissue (nalt) and thus claudin-4-targeting using c-terminal fragment of clostridium perfringens enterotoxin (c-cpe) effectively delivered fused antigen to nalt and consequently induced antigen-specific immune responses. in this study, we applie ...201526018248
critical illness in pregnancy: part ii: common medical conditions complicating pregnancy and puerperium.the first of this two-part series on critical illness in pregnancy dealt with obstetric disorders. in part ii, medical conditions that commonly affect pregnant women or worsen during pregnancy are discussed. ards occurs more frequently in pregnancy. strategies commonly used in nonpregnant patients, including permissive hypercapnia, limits for plateau pressure, and prone positioning, may not be acceptable, especially in late pregnancy. genital tract infections unique to pregnancy include chorioam ...201526020727
acute cholecystitis associated with infection of enterobacteriaceae from gut microbiota.acute cholecystitis (ac) is one of the most common surgical diseases. bacterial infection accounts for 50% to 85% of the disease's onset. since there is a close relationship between the biliary system and the gut, the aims of this study were to characterize and determine the influence of gut microbiota on ac, to detect the pathogenic microorganism in the biliary system, and to explore the relationship between the gut and bile microbiota of patients with ac. a total of 185 713 high-quality sequen ...201526025761
lipoproteins from clostridium perfringens and their protective efficacy in mouse model.clostridium perfringens is an obligately anaerobic rod-shaped bacterium and etiological agent for several diseases in humans and animals. the pathogen has been listed as validated biological agent and warrants development of medical countermeasures. the homologs of some of the lipoproteins identified from various fractions of c. perfringens in our previous studies were observed to be virulence determinants in other pathogenic bacteria. three putative virulence associated lipoproteins; polysaccha ...201526027922
an internal thioester in a pathogen surface protein mediates covalent host binding.to cause disease and persist in a host, pathogenic and commensal microbes must adhere to tissues. colonization and infection depend on specific molecular interactions at the host-microbe interface that involve microbial surface proteins, or adhesins. to date, adhesins are only known to bind to host receptors non-covalently. here we show that the streptococcal surface protein sfbi mediates covalent interaction with the host protein fibrinogen using an unusual internal thioester bond as a 'chemica ...201526032562
claudin-binder c-cpe mutants enhance permeability of insulin across human nasal epithelial cells.intranasal insulin administration has therapeutic potential for alzheimer's disease and in intranasal administration across the nasal mucosa, the paracellular pathway regulated by tight junctions is important. the c-terminal fragment of clostridium perfringens enterotoxin (c-cpe) binds the tight junction protein claudin and disrupts the tight junctional barrier without a cytotoxic effect. the c-cpe mutant called c-cpe 194 binds only to claudin-4, whereas the c-cpe 194 mutant called c-cpe m19 bin ...201626036653
hydrolyzable and condensed tannins resistance in clostridium perfringens.tannins added in the diet are being used to improve nutrition and health in farm animals as an alternative to antibiotic growth promoters and to control enteric clostridial diseases. however, the capacity of clostridium perfringens to develop resistance under the selective pressure of tannins is unknown. the purpose of this study was to determine if c. perfringens possess the ability to develop resistance against tannins in comparison with antimicrobial agents. susceptibility for 7 agps (antimic ...201526037239
impact of a drug-free program on broiler chicken growth performances, gut health, clostridium perfringens and campylobacter jejuni occurrences at the farm level.the use of antimicrobial agents as feed additives in poultry production is a public health concern due to the overall increase in antimicrobial resistance. although some alternative products are commercially available, little is known on their potential impact on flock health and productivity. a prospective study involving 1.55 million birds was conducted on eight commercial broiler farms in québec, canada, to evaluate the impact of replacing antibiotic growth promoters and anticoccidial drugs b ...201526047674
fatal clostridium perfringens septicemia suggested by postmortem computed tomography: a medico-legal autopsy case report.we report a fatal case of suspected clostridium (cl.) perfringens septicemia in a previously healthy woman in her eighties. at first, she presented at the hospital complaining of upper abdominal discomfort and vomiting, and was discharged the next day after ruling out any fatal conditions. however, her condition deteriorated approximately 10h after discharge and she died shortly after. the postmortem computed tomography (pmct) performed 29h postmortem revealed an excessive systemic gas accumulat ...201526048862
identification of the replication region in pbcnf5603, a bacteriocin-encoding plasmid, in the enterotoxigenic clostridium perfringens strain f5603.most recent studies of clostridium perfringens plasmids have focused on toxin-encoding or antibiotic resistance plasmids. to cause intestinal disease, a toxigenic strain must grow in the intestines to levels allowing for sufficient toxin production and this in vivo growth often involves overcoming the normal intestinal microbial population. for this purpose, bacteriocin production might be important.201526055257
erratum for maheux et al., abilities of the mcp agar method and crename alpha toxin-specific real-time pcr assay to detect clostridium perfringens spores in drinking water. 201426058088
detailed characterization of the o-linked glycosylation of the neuropilin-1 c/mam-domain.neuropilins are involved in angiogenesis and neuronal development. the membrane proximal domain of neuropilin-1, called c or mam domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. the c/mam domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. we found that the c/mam domain wa ...201626059692
clostridium perfringens enterotoxin c-terminal domain labeled to fluorescent dyes for in vivo visualization of micrometastatic chemotherapy-resistant ovarian cancer.identification of micrometastatic disease at the time of surgery remains extremely challenging in ovarian cancer patients. we used fluorescence microscopy, an in vivo imaging system and a fluorescence stereo microscope to evaluate fluorescence distribution in claudin-3- and -4-overexpressing ovarian tumors, floating tumor clumps isolated from ascites and healthy organs. to do so, mice harboring chemotherapy-naïve and chemotherapy-resistant human ovarian cancer xenografts or patient-derived xenog ...201526060989
convergent weaponry in a biological arms race.bacterial surface proteins covalently attach to host cells via a mechanism that is also used by immune system proteins that help eliminate invading pathogens.201526062771
oncoleaking: use of the pore-forming clostridium perfringens enterotoxin (cpe) for suicide gene therapy.suicide gene therapy has been shown to be very efficient in tumor eradication. numerous suicide genes were tested in vitro and in vivo demonstrating their therapeutic potential in clinical trials. apart from this, still growing efforts are made to generate more targeted and more effective suicide gene systems for cancer gene therapy. in this regard bacterial toxins are an alternative, which add to the broad spectrum of different suicide strategies. in this context, the claudin-targeted bacterial ...201526072402
a thermophilic phage endolysin fusion to a clostridium perfringens-specific cell wall binding domain creates an anti-clostridium antimicrobial with improved thermostability.clostridium perfringens is the third leading cause of human foodborne bacterial disease and is the presumptive etiologic agent of necrotic enteritis among chickens. treatment of poultry with antibiotics is becoming less acceptable. endolysin enzymes are potential replacements for antibiotics. many enzymes are added to animal feed during production and are subjected to high-heat stress during feed processing. to produce a thermostabile endolysin for treating poultry, an e. coli codon-optimized ge ...201526075507
cholesterol segregates into submicrometric domains at the living erythrocyte membrane: evidence and regulation.although cholesterol is essential for membrane fluidity and deformability, the level of its lateral heterogeneity at the plasma membrane of living cells is poorly understood due to lack of appropriate probe. we here report on the usefulness of the d4 fragment of clostridium perfringens toxin fused to mcherry (theta*), as specific, non-toxic, sensitive and quantitative cholesterol-labeling tool, using erythrocyte flat membrane. by confocal microscopy, theta* labels cholesterol-enriched submicrome ...201526077601
clostridium perfringens epsilon toxin causes selective death of mature oligodendrocytes and central nervous system demyelination.clostridium perfringens epsilon toxin (ε-toxin) is responsible for a devastating multifocal central nervous system (cns) white matter disease in ruminant animals. the mechanism by which ε-toxin causes white matter damage is poorly understood. in this study, we sought to determine the molecular and cellular mechanisms by which ε-toxin causes pathological changes to white matter. in primary cns cultures, ε-toxin binds to and kills oligodendrocytes but not astrocytes, microglia, or neurons. in cere ...201526081637
using broiler litter and swine manure lagoon effluent in sawdust-based swine mortality composts: effects on nutrients, bacteria, and gaseous emissions.disposition of mortalities challenges confined animal feeding operations (cafos), especially sow (farrowing) farms, which experience mortalities daily. regulations and transportation costs may preclude incineration, landfill burial, and rendering; therefore, swine cafos in mississippi in the mid-south u.s. often compost mortalities. in this study, a farm-standard composting mix of sawdust (s) and water (w) was compared with mixes where n was supplied by broiler litter (l) and water was replaced ...201526081729
plant-derived monoclonal antibodies for prevention and treatment of infectious disease.numerous monoclonal antibodies (mabs) that recognize and neutralize infectious pathogens have been isolated and developed over the years. the fact that infectious diseases can involve large populations of infected individuals is an important factor that has motivated the search for both cost-effective and scalable methods of antibody production. the current technologies for production of antibodies in plants allow for very rapid expression and evaluation that can also be readily scaled for multi ...201426082108
gas-forming pyogenic liver abscess with septic shock.the pyogenic liver abscess caused by clostridium perfringens (c. perfringens) is a rare but rapidly fatal infection. the main virulence factor of this pathogen is its α-toxin (lecithinase), which decomposes the phospholipid in cell membranes leading to cell lysis. once the bacteria are in blood stream, massive intravascular hemolysis occurs. this can present as anemia on admission with evidence of hemolysis as indicated by low serum haptoglobin, high serum lactate dehydrogenase (ldh), elevated i ...201526090240
virulence plasmids of spore-forming bacteria.plasmid-encoded virulence factors are important in the pathogenesis of diseases caused by spore-forming bacteria. unlike many other bacteria, the most common virulence factors encoded by plasmids in clostridium and bacillus species are protein toxins. clostridium perfringens causes several histotoxic and enterotoxin diseases in both humans and animals and produces a broad range of toxins, including many pore-forming toxins such as c. perfringens enterotoxin, epsilon-toxin, beta-toxin, and netb. ...201426104459
genome wide transcriptomic analysis identifies pathways affected by the infusion of clostridium perfringens culture supernatant in the duodenum of broilers in situ.clostridium perfringens type a is the main etiological factor for necrotic enteritis, a multifactorial enteric disease that penalizes performance, health, and welfare of poultry. lack of knowledge of host responses and disease pathogenesis is slowing down progress on developing therapies for disease control. a combined genomewide and targeted gene approach was used to investigate pathways and biological functions affected by the infusion of c. perfringens culture supernatant in the duodenum of b ...201526115301
differential decay of wastewater bacteria and change of microbial communities in beach sand and seawater microcosms.laboratory microcosm experiments were conducted to determine the decay kinetics of wastewater bacteria and the change of microbial communities in beach sand and seawater. cultivation-based methods showed that common fecal indicator bacteria (fibs; escherichia coli, enterococci, and clostridium perfringens) exhibited biphasic decay patterns in all microcosms. enterococci and c. perfringens, but not e. coli, showed significantly smaller decay rates in beach sand than in seawater. cultivation-indep ...201526125493
carriage of clostridium perfringens by benthic crabs in a sewage-polluted estuary.the estuary of bahía blanca (ebb), argentina, is an important wetland under intense sewage pollution. we investigated the occurrence of clostridium perfringens (cp) in populations of two benthic crabs (neohelice granulata and cyrtograpsus angulatus) and in sediment from the ebb. cp was found in 49.1% of the crabs and all of the isolates were identified as type a. the alpha (cpa) and enterotoxin (cpe) encoding genes were identified. genetic analyses identified 13 novel sequence types, and found n ...201526130524
in vitro probiotic characteristics of lactobacillus plantarum zdy 2013 and its modulatory effect on gut microbiota of mice.lactobacillus plantarum zdy 2013, a novel strain isolated from chinese traditional fermented acid beans, was systematically evaluated for its survival capacity under stress conditions (ph, bile salt, simulated gastrointestinal tract, and antibiotics), production of exopolysaccharide and antagonism against 8 pathogens. its effect on mice gut microbiota was also investigated by quantitative pcr and pcr-denaturing gradient gel electrophoresis. the results showed that zdy 2013 can grow at ph 3.5 and ...201526142853
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