Sample records for clostridium perfringens epsilon

  1. Effects of Clostridium perfringens alpha and epsilon toxins in the bovine gut.

    PubMed

    Morris, Winston E; Dunleavy, Mariana V; Diodati, Julián; Berra, Guillermo; Fernandez-Miyakawa, Mariano E

    2012-02-01

    Clostridium perfringens alpha and epsilon toxins produce enterotoxaemia in sheep and goats. However, the information regarding the pathophysiology of alpha and epsilon toxins in the bovine intestine is still scanty. In this study, intestinal loops were performed in the ileum and colon of three one-week-old Holstein and two four-week-old crossbreed calves. Laparotomy was performed in all calves under anaesthesia and four loops -three cm long- were performed in the small and large intestines. For both intestines, loops were inoculated with alpha or epsilon toxins. Tissue samples from all loops were obtained and processed for routine histology and for transmission electron microscopy. Congestion was observed in toxin treated loops. Fluid accumulation in the gut lumen was prominent in all treated loops, but in epsilon treated ones the mucous was also haemorrhagic. The histology revealed large amount of exfoliated epithelial cells in the lumen of alpha toxin treated loops and severe haemorrhage was observed in the lamina propria of epsilon toxin treated colonic loops. Despite some necrotic exfoliated enterocytes, no ultraestructural changes were observed in alpha toxin treated loops, though with epsilon toxin the loops exhibited dilation of the intercellular space in the mucosa of both, small and large intestines. These observations indicate that both, alpha and epsilon toxins can alter the intestinal barrier, in calves and are pathogenic for this species. PMID:22178571

  2. CLOSTRIDIUM PERFRINGENS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The incidence of C. perfringens food-poisoning is quite common and costly. Although somewhat fastidious in growth characteristics using synthetic laboratory media, the microorganism is very prolific when found in food products. Despite the pathogen’s ubiquity in the natural environment, foodborne i...

  3. Clostridium perfringens Epsilon Toxin Causes Selective Death of Mature Oligodendrocytes and Central Nervous System Demyelination

    PubMed Central

    Linden, Jennifer R.; Ma, Yinghua; Zhao, Baohua; Harris, Jason Michael; Rumah, Kareem Rashid; Schaeren-Wiemers, Nicole

    2015-01-01

    ABSTRACT Clostridium perfringens epsilon toxin (?-toxin) is responsible for a devastating multifocal central nervous system (CNS) white matter disease in ruminant animals. The mechanism by which ?-toxin causes white matter damage is poorly understood. In this study, we sought to determine the molecular and cellular mechanisms by which ?-toxin causes pathological changes to white matter. In primary CNS cultures, ?-toxin binds to and kills oligodendrocytes but not astrocytes, microglia, or neurons. In cerebellar organotypic culture, ?-toxin induces demyelination, which occurs in a time- and dose-dependent manner, while preserving neurons, astrocytes, and microglia. ?-Toxin specificity for oligodendrocytes was confirmed using enriched glial culture. Sensitivity to ?-toxin is developmentally regulated, as only mature oligodendrocytes are susceptible to ?-toxin; oligodendrocyte progenitor cells are not. ?-Toxin sensitivity is also dependent on oligodendrocyte expression of the proteolipid myelin and lymphocyte protein (MAL), as MAL-deficient oligodendrocytes are insensitive to ?-toxin. In addition, ?-toxin binding to white matter follows the spatial and temporal pattern of MAL expression. A neutralizing antibody against ?-toxin inhibits oligodendrocyte death and demyelination. This study provides several novel insights into the action of ?-toxin in the CNS. (i) ?-Toxin causes selective oligodendrocyte death while preserving all other neural elements. (ii) ?-Toxin-mediated oligodendrocyte death is a cell autonomous effect. (iii) The effects of ?-toxin on the oligodendrocyte lineage are restricted to mature oligodendrocytes. (iv) Expression of the developmentally regulated proteolipid MAL is required for the cytotoxic effects. (v) The cytotoxic effects of ?-toxin can be abrogated by an ?-toxin neutralizing antibody. PMID:26081637

  4. Epsilon toxin from Clostridium perfringens acts on oligodendrocytes without forming pores, and causes demyelination.

    PubMed

    Wioland, Laetitia; Dupont, Jean-Luc; Doussau, Frédéric; Gaillard, Stéphane; Heid, Flavia; Isope, Philippe; Pauillac, Serge; Popoff, Michel R; Bossu, Jean-Louis; Poulain, Bernard

    2015-03-01

    Epsilon toxin (ET) is produced by Clostridium perfringens types B and D and causes severe neurological disorders in animals. ET has been observed binding to white matter, suggesting that it may target oligodendrocytes. In primary cultures containing oligodendrocytes and astrocytes, we found that ET (10(-9) M and 10(-7) M) binds to oligodendrocytes, but not to astrocytes. ET induces an increase in extracellular glutamate, and produces oscillations of intracellular Ca(2+) concentration in oligodendrocytes. These effects occurred without any change in the transmembrane resistance of oligodendrocytes, underlining that ET acts through a pore-independent mechanism. Pharmacological investigations revealed that the Ca(2+) oscillations are caused by the ET-induced rise in extracellular glutamate concentration. Indeed, the blockade of metabotropic glutamate receptors type 1 (mGluR1) prevented ET-induced Ca(2+) signals. Activation of the N-methyl-D-aspartate receptor (NMDA-R) is also involved, but to a lesser extent. Oligodendrocytes are responsible for myelinating neuronal axons. Using organotypic cultures of cerebellar slices, we found that ET induced the demyelination of Purkinje cell axons within 24 h. As this effect was suppressed by antagonizing mGluR1 and NMDA-R, demyelination is therefore caused by the initial ET-induced rise in extracellular glutamate concentration. This study reveals the novel possibility that ET can act on oligodendrocytes, thereby causing demyelination. Moreover, it suggests that for certain cell types such as oligodendrocytes, ET can act without forming pores, namely through the activation of an undefined receptor-mediated pathway. PMID:25287162

  5. Proteolytic Processing and Activation of Clostridium perfringens Epsilon Toxin by Caprine Small Intestinal Contents

    PubMed Central

    Freedman, John C.; Li, Jihong; Uzal, Francisco A.

    2014-01-01

    ABSTRACT Epsilon toxin (ETX), a pore-forming toxin produced by type B and D strains of Clostridium perfringens, mediates severe enterotoxemia in livestock and possibly plays a role in human disease. During enterotoxemia, the nearly inactive ETX prototoxin is produced in the intestines but then must be activated by proteolytic processing. The current study sought to examine ETX prototoxin processing and activation ex vivo using the intestinal contents of a goat, a natural host species for ETX-mediated disease. First, this study showed that the prototoxin has a KEIS N-terminal sequence with a molecular mass of 33,054 Da. When the activation of ETX prototoxin ex vivo by goat small intestinal contents was assessed by SDS-PAGE, the prototoxin was processed in a stepwise fashion into an ~27-kDa band or higher-molecular-mass material that could be toxin oligomers. Purified ETX corresponding to the ~27-kDa band was cytotoxic. When it was biochemically characterized by mass spectrometry, the copresence of three ETX species, each with different C-terminal residues, was identified in the purified ~27-kDa ETX preparation. Cytotoxicity of each of the three ETX species was then demonstrated using recombinant DNA approaches. Serine protease inhibitors blocked the initial proteotoxin processing, while carboxypeptidase inhibitors blocked further processing events. Taken together, this study provides important new insights indicating that, in the intestinal lumen, serine protease (including trypsin and possibly chymotrypsin) initiates the processing of the prototoxin but other proteases, including carboxypeptidases, then process the prototoxin into multiple active and stable species. PMID:25336460

  6. Distribution of Clostridium perfringens epsilon toxin in the brains of acutely intoxicated mice and its effect upon glial cells.

    PubMed

    Soler-Jover, Alex; Dorca, Jonatan; Popoff, Michel R; Gibert, Maryse; Saura, Josep; Tusell, Josep Maria; Serratosa, Joan; Blasi, Juan; Martín-Satué, Mireia

    2007-09-15

    Epsilon toxin (epsilon-toxin), produced by Clostridium perfringens types B and D, causes fatal enterotoxaemia in livestock. The disease is principally manifested as severe and often fatal neurological disturbance. Oedema of several organs, including the brain, is also a clinical sign related to microvascular damage. Recombinant epsilon-toxin-green fluorescence protein (epsilon-toxin-GFP) and epsilon-prototoxin-GFP have already been characterised as useful tools to track their distribution in intravenously injected mice, by means of direct fluorescence microscopy detection. The results shown here, using an acutely intoxicated mouse model, strongly suggest that epsilon-toxin-GFP, but not epsilon-prototoxin-GFP, not only causes oedema but is also able to cross the blood-brain barrier and accumulate in brain tissue. In some brain areas, epsilon-toxin-GFP is found bound to glial cells, both astrocytes and microglia. Moreover, cytotoxicity assays, performed with mixed glial primary cultures, demonstrate the cytotoxic effect of epsilon-toxin upon both astrocytes and microglial cells. PMID:17572464

  7. Clostridium botulinum and Clostridium perfringens

    Microsoft Academic Search

    Jim McLauchlin; Kathie A. Grant

    \\u000a Clostridium is a diverse genus of Gram-positive, endospore-bearing obligate anaerobes that are widespread in the environment. This genus\\u000a includes more than 100 species, and the overall range in the G+C content (22–55 mol%) reflects the enormous phylogenetic variation\\u000a encompassed within this group. The principal foodborne pathogens are Clostridium botulinum and Clostridium perfringens that cause toxin-mediated disease either by preformed toxin

  8. Clostridium perfringens Epsilon Toxin Targets Granule Cells in the Mouse Cerebellum and Stimulates Glutamate Release

    PubMed Central

    Lonchamp, Etienne; Dupont, Jean-Luc; Wioland, Laetitia; Courjaret, Raphaël; Mbebi-Liegeois, Corinne; Jover, Emmanuel; Doussau, Frédéric; Popoff, Michel R.; Bossu, Jean-Louis; de Barry, Jean; Poulain, Bernard

    2010-01-01

    Epsilon toxin (ET) produced by C. perfringens types B and D is a highly potent pore-forming toxin. ET-intoxicated animals express severe neurological disorders that are thought to result from the formation of vasogenic brain edemas and indirect neuronal excitotoxicity. The cerebellum is a predilection site for ET damage. ET has been proposed to bind to glial cells such as astrocytes and oligodendrocytes. However, the possibility that ET binds and attacks the neurons remains an open question. Using specific anti-ET mouse polyclonal antibodies and mouse brain slices preincubated with ET, we found that several brain structures were labeled, the cerebellum being a prominent one. In cerebellar slices, we analyzed the co-staining of ET with specific cell markers, and found that ET binds to the cell body of granule cells, oligodendrocytes, but not astrocytes or nerve endings. Identification of granule cells as neuronal ET targets was confirmed by the observation that ET induced intracellular Ca2+ rises and glutamate release in primary cultures of granule cells. In cultured cerebellar slices, whole cell patch-clamp recordings of synaptic currents in Purkinje cells revealed that ET greatly stimulates both spontaneous excitatory and inhibitory activities. However, pharmacological dissection of these effects indicated that they were only a result of an increased granule cell firing activity and did not involve a direct action of the toxin on glutamatergic nerve terminals or inhibitory interneurons. Patch-clamp recordings of granule cell somata showed that ET causes a decrease in neuronal membrane resistance associated with pore-opening and depolarization of the neuronal membrane, which subsequently lead to the firing of the neuronal network and stimulation of glutamate release. This work demonstrates that a subset of neurons can be directly targeted by ET, suggesting that part of ET-induced neuronal damage observed in neuronal tissue is due to a direct effect of ET on neurons. PMID:20941361

  9. Toxin Plasmids of Clostridium perfringens

    PubMed Central

    Li, Jihong; Adams, Vicki; Bannam, Trudi L.; Miyamoto, Kazuaki; Garcia, Jorge P.; Uzal, Francisco A.; Rood, Julian I.

    2013-01-01

    SUMMARY In both humans and animals, Clostridium perfringens is an important cause of histotoxic infections and diseases originating in the intestines, such as enteritis and enterotoxemia. The virulence of this Gram-positive, anaerobic bacterium is heavily dependent upon its prolific toxin-producing ability. Many of the ?16 toxins produced by C. perfringens are encoded by large plasmids that range in size from ?45 kb to ?140 kb. These plasmid-encoded toxins are often closely associated with mobile elements. A C. perfringens strain can carry up to three different toxin plasmids, with a single plasmid carrying up to three distinct toxin genes. Molecular Koch's postulate analyses have established the importance of several plasmid-encoded toxins when C. perfringens disease strains cause enteritis or enterotoxemias. Many toxin plasmids are closely related, suggesting a common evolutionary origin. In particular, most toxin plasmids and some antibiotic resistance plasmids of C. perfringens share an ?35-kb region containing a Tn916-related conjugation locus named tcp (transfer of clostridial plasmids). This tcp locus can mediate highly efficient conjugative transfer of these toxin or resistance plasmids. For example, conjugative transfer of a toxin plasmid from an infecting strain to C. perfringens normal intestinal flora strains may help to amplify and prolong an infection. Therefore, the presence of toxin genes on conjugative plasmids, particularly in association with insertion sequences that may mobilize these toxin genes, likely provides C. perfringens with considerable virulence plasticity and adaptability when it causes diseases originating in the gastrointestinal tract. PMID:23699255

  10. Amino acid residue Y196E substitution and C-terminal peptide synergistically alleviate the toxicity of Clostridium perfringens epsilon toxin.

    PubMed

    Yao, Wenwu; Kang, Lin; Gao, Shan; Zhuang, Xiangjin; Zhang, Tao; Yang, Hao; Ji, Bin; Xin, Wenwen; Wang, Jinglin

    2015-06-15

    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-terminal peptides and 23 C-terminal peptides), rETX-C (rETX with 23 C-terminal peptides), rETX(Y196E) (rETX with an amino acid residue substitution at Y196) and rETX(Y196E)-C (rETX-C with a Y196E mutation), were constructed in this study. Both the amino acid residue Y196E substitution and the C-terminal peptide reduce ETX toxicity to a similar extent, and the two factors synergistically alleviate ETX toxicity. In addition, we demonstrated that the C-terminal peptides and Y196E amino acid mutation reduce the toxin toxicity in two different pathways: the C-terminal peptides inhibit the binding activity of toxins to target cells, and the Y196E amino acid mutation slightly inhibits the pore-forming or heptamer-forming process. Interaction between the two factors was not observed in pore-forming or binding assays but toxicity assays, which demonstrated that the relationship between domains of the toxin is more complicated than previously appreciated. However, the exact mechanism of synergistic action is not yet clarified. PMID:25912943

  11. Enterotoxin Plasmid from Clostridium perfringens Is Conjugative

    Microsoft Academic Search

    SIGRID BRYNESTAD; MAHFUZUR R. SARKER; BRUCE A. MCCLANE; PER EINAR GRANUM; JULIAN I. ROOD

    2001-01-01

    Clostridium perfringens enterotoxin is the major virulence factor involved in the pathogenesis of C. perfringens type A food poisoning and several non-food-borne human gastrointestinal illnesses. The enterotoxin gene, cpe, is located on the chromosome of food-poisoning isolates but is found on a large plasmid in non-food-borne gastrointestinal disease isolates and in veterinary isolates. To evaluate whether the cpe plasmid encodes

  12. Clostridium perfringens : A Dynamic Foodborne Pathogen

    Microsoft Academic Search

    Santos García; Norma Heredia

    2011-01-01

    Clostridium perfringens is a spore-forming bacterium and natural inhabitant of soil and the intestinal tracts of many warm-blooded animals, including\\u000a humans. The ubiquitous nature of this bacterium and its spores makes it a frequent problem for the food industry and establishments\\u000a where large amounts of food are prepared. C. perfringens causes potentially lethal foodborne diseases in humans, including food poisoning

  13. Comparative Analysis of Clostridium perfringens Bacteriophage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium perfringens are Gram-positive bacteria that are a major bacterial cause of food-borne disease among humans. These anaerobic bacteria are also the presumptive etiologic agent of necrotic enteritis among chickens. Pathogenesis and symptoms of a necrotic enteritis infection among chickens ...

  14. Clostridium perfringens in Meat and Meat Products

    PubMed Central

    Hall, Herbert E.; Angelotti, Robert

    1965-01-01

    A total of 262 specimens of meat and meat dishes were examined for the presence of Clostridium perfringens. Of this total, 161 were raw, unprocessed beef, veal, lamb, pork, or chicken; 101 were processed meats and meat dishes. C. perfringens was isolated from 113 (43.1%) of these specimens. The highest percentage of contamination (82%) was found in veal cuts, and the lowest (4.7%) in sliced sandwich meats and spreads. Only 2 of the 113 isolates were shown to produce heat-resistant spores, which indicates a very low incidence (0.8%) of contamination. These findings indicate that outbreaks of C. perfringens food-borne disease in the Cincinnati area are caused principally by the contamination of the food with vegetative cells or spores of the organism after cooking. Studies of the effects of various holding temperatures on the growth of C. perfringens indicated that, in the range of 5 to 15 C, no multiplication would occur, but that viable cells would still be present at the end of a 5-day holding period. Extremely rapid growth occurred at temperatures around 45 C, and complete inhibition of growth was accomplished between 49 and 52 C. PMID:14325274

  15. Clostridium perfringens infection after transarterial chemoembolization for large hepatocellular carcinoma.

    PubMed

    Li, Jing-Huan; Yao, Rong-Rong; Shen, Hu-Jia; Zhang, Lan; Xie, Xiao-Ying; Chen, Rong-Xin; Wang, Yan-Hong; Ren, Zheng-Gang

    2015-04-14

    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 the liver and renal function turned normal. The 2(nd) TACE procedure was performed 1.5 mo later and no recurrent Clostridium perfringens infection was found. PMID:25892893

  16. Clostridium perfringens infection after transarterial chemoembolization for large hepatocellular carcinoma

    PubMed Central

    Li, Jing-Huan; Yao, Rong-Rong; Shen, Hu-Jia; Zhang, Lan; Xie, Xiao-Ying; Chen, Rong-Xin; Wang, Yan-Hong; Ren, Zheng-Gang

    2015-01-01

    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 2nd 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 the liver and renal function turned normal. The 2nd TACE procedure was performed 1.5 mo later and no recurrent Clostridium perfringens infection was found. PMID:25892893

  17. Perfringolysin O: The Underrated Clostridium perfringens Toxin?

    PubMed Central

    Verherstraeten, Stefanie; Goossens, Evy; Valgaeren, Bonnie; Pardon, Bart; Timbermont, Leen; Haesebrouck, Freddy; Ducatelle, Richard; Deprez, Piet; Wade, Kristin R.; Tweten, Rodney; Van Immerseel, Filip

    2015-01-01

    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 ? 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 surface. The prepore then undergoes conversion into the bilayer-spanning pore measuring approximately 250–300 Å in diameter. PFO is expressed in nearly all identified C. perfringens strains and harbors interesting traits that suggest a potential undefined role for PFO in disease development. Research has demonstrated a role for PFO in gas gangrene progression and bovine necrohemorrhagic enteritis, but there is limited data available to determine if PFO also functions in additional disease presentations caused by C. perfringens. This review summarizes the known structural and functional characteristics of PFO, while highlighting recent insights into the potential contributions of PFO to disease pathogenesis. PMID:26008232

  18. Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates

    Microsoft Academic Search

    Christoph G Baums; Ulrich Schotte; Gunter Amtsberg; Ralph Goethe

    2004-01-01

    In this study we provide a protocol for genotyping Clostridium perfringens with a new multiplex PCR. This PCR enables reliable and specific detection of the toxin genes cpa, cpb, etx, iap, cpe and cpb2 from heat lysed bacterial suspensions. The efficiency of the protocol was demonstrated by typing C. perfringens reference strains and isolates from veterinary bacteriological routine diagnostic specimens.

  19. Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates.

    PubMed

    Baums, Christoph G; Schotte, Ulrich; Amtsberg, Gunter; Goethe, Ralph

    2004-05-20

    In this study we provide a protocol for genotyping Clostridium perfringens with a new multiplex PCR. This PCR enables reliable and specific detection of the toxin genes cpa, cpb, etx, iap, cpe and cpb2 from heat lysed bacterial suspensions. The efficiency of the protocol was demonstrated by typing C. perfringens reference strains and isolates from veterinary bacteriological routine diagnostic specimens. PMID:15135508

  20. [Clostridium perfringens septicemia associated with foodborne toxic infection and abortion].

    PubMed

    Lantelme, P; Mohammedi, I; Duperret, S; Vedrinne, J M; Allaouchiche, B; Motin, J

    1995-01-01

    A 32-year-old pregnant woman with poor life and hygiene conditions presented with premature labour, fever and diarrhoea. After admission she gave birth to a stillborn child. The examination revealed a septicaemia with massive haemolysis and renal failure. Six blood cultures obtained on admission yielded Clostridium perfringens. The outcome was favourable after an adapted antibiomicrobial therapy. This case illustrates the potential severity of Clostridium perfringens foodborne toxi-infection which can lead to abortion and septicaemia with massive haemolysis. PMID:8572393

  1. Animal models to study the pathogenesis of human and animal Clostridium perfringens infections.

    PubMed

    Uzal, Francisco A; McClane, Bruce A; Cheung, Jackie K; Theoret, James; Garcia, Jorge P; Moore, Robert J; Rood, Julian I

    2015-08-31

    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 to the identification of NetB as the primary toxin involved in disease. C. perfringens type A food poisoning is a highly prevalent bacterial illness in the USA and elsewhere. Rabbits and mice are the species most commonly used to study the action of enterotoxin, the causative toxin. Other animal models used to study the effect of this toxin are rats, non-human primates, sheep and cattle. In rabbits and mice, CPE produces severe necrosis of the small intestinal epithelium along with fluid accumulation. C. perfringens type D infection has been studied by inoculating epsilon toxin (ETX) intravenously into mice, rats, sheep, goats and cattle, and by intraduodenal inoculation of whole cultures of this microorganism in mice, sheep, goats and cattle. Molecular Koch's postulates have been fulfilled for enterotoxigenic C. perfringens type A in rabbits and mice, for C. perfringens type A necrotic enteritis and gas gangrene in chickens and mice, respectively, for C. perfringens type C in mice, rabbits and goats, and for C. perfringens type D in mice, sheep and goats. PMID:25770894

  2. Toxin types of Clostridium perfringens isolated from free-ranging, semi-domesticated reindeer in Norway.

    PubMed

    Aschfalk, A; Valentin-Weigand, P; Müller, W; Goethe, R

    2002-08-17

    Samples of faeces were taken from 166 healthy domesticated reindeer (Rangifer tarandus tarandus) from three flocks in different reindeer husbandry districts in northern Norway and examined bacteriologically for the presence of Clostridium perfringens. The organism was isolated from 98 (59 per cent) of the reindeer. The isolates were classified into C perfringens toxin types by PCR analysis specific for the genes encoding the four major toxins (alpha, beta, epsilon and tau) and were subclassified by the detection of the genes encoding C perfringens beta2-toxin and enterotoxin. All the isolates belonged to C perfringens toxin type A. In addition, 15 of the 98 isolates were PCR-positive for the beta2-toxin gene, and two of the isolates had the the gene encoding for enterotoxin. PMID:12211393

  3. Toxigenic Clostridium perfringens from a Parvovirus-Infected Dog

    PubMed Central

    Tilton, R. C.; Van Kruiningen, H. J.; Kwasnik, I.; Ryan, R. W.

    1981-01-01

    A strain of Clostridium perfringens, type A, has been isolated from the intestine of a dog which died from parvovirus infection. This Clostridium strain produces a toxin which can be detected by counterimmunoelectrophoresis, using C. difficile antitoxin, and produces cytotoxicity in WI-38 cell culture. Cytopathology can be blocked by C. difficile antitoxin. Its role in canine parvovirus infection is unknown. PMID:6277989

  4. Toxigenic Clostridium perfringens from a parvovirus-infected dog.

    PubMed

    Tilton, R C; Van Kruiningen, H J; Kwasnik, I; Ryan, R W

    1981-12-01

    A strain of Clostridium perfringens, type A, has been isolated from the intestine of a dog which died from parvovirus infection. This Clostridium strain produces a toxin which can be detected by counterimmunoelectrophoresis, using C. difficile antitoxin, and produces cytotoxicity in WI-38 cell culture. Cytopathology can be blocked by C. difficile antitoxin. Its role in canine parvovirus infection is unknown. PMID:6277989

  5. Characterization of Clostridium perfringens in the feces of adult horses and foals with acute enterocolitis

    PubMed Central

    Gohari, Iman Mehdizadeh; Arroyo, Luis; MacInnes, Janet I.; Timoney, John F.; Parreira, Valeria R.; Prescott, John F.

    2014-01-01

    Up to 60% of cases of equine colitis have no known cause. To improve understanding of the causes of acute colitis in horses, we hypothesized that Clostridium perfringens producing enterotoxin (CPE) and/or beta2 toxin (CPB2) are common and important causes of severe colitis in horses and/or that C. perfringens producing an as-yet-undescribed cytotoxin may also cause colitis in horses. Fecal samples from 55 horses (43 adults, 12 foals) with clinical evidence of colitis were evaluated by culture for the presence of Clostridium difficile, C. perfringens, and Salmonella. Feces were also examined by enzyme-linked immunosorbent assay (ELISA) for C. difficile A/B toxins and C. perfringens alpha toxin (CPA), beta2 toxin (CPB2), and enterotoxin (CPE). Five C. perfringens isolates per sample were genotyped for the following genes: cpa, cpb, cpb2 consensus, cpb2 atypical, cpe (enterotoxin), etx (epsilon toxin), itx (iota toxin), netB (necrotic enteritis toxin B), and tpeL (large C. perfringens cytotoxin). The supernatants of these isolates were also evaluated for toxicity for an equine cell line. All fecal samples were negative for Salmonella. Clostridium perfringens and C. difficile were isolated from 40% and 5.4% of samples, respectively. All fecal samples were negative for CPE. Clostridium perfringens CPA and CPB2 toxins were detected in 14.5% and 7.2% of fecal samples, respectively, all of which were culture-positive for C. perfringens. No isolates were cpe, etx, netB, or tpeL gene-positive. Atypical cpb2 and consensus cpb2 genes were identified in 15 (13.6%) and 4 (3.6%) of 110 isolates, respectively. All equine C. perfringens isolates showed far milder cytotoxicity effects than a CPB-producing positive control, although cpb2-positive isolates were slightly but significantly more cytotoxic than negative isolates. Based on this studied population, we were unable to confirm our hypothesis that CPE and CPB2-producing C. perfringens are common in horses with colitis in Ontario and we failed to identify cytotoxic activity in vitro in the type A isolates recovered. PMID:24396174

  6. MATHEMATICAL MODELING OF GROWTH OF CLOSTRIDIUM PERFRINGENS IN COOKED BEEF

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this work was to study the growth kinetics of Clostridium perfringens spores in thermally processed ground beef and compare the suitability of the Gompertz, logistic, and Baranyi models used to describe the isothermal bacterial growth. Ground beef samples inoculated with the spores ...

  7. Tips to Prevent Illness from Clostridium Perfringens

    MedlinePLUS

    ... estimated to cause nearly 1 million cases of foodborne illness (sometimes called "food poisoning") each year. C. perfringens is found in many environmental sources as well as in the intestines of humans and animals. What are common food sources of C. perfringens ? ...

  8. Antimicrobial susceptibility of Clostridium perfringens strains isolated from broiler chickens

    PubMed Central

    Silva, R. O. S.; Salvarani, F.M.; Assis, R.A.; Martins, N.R.S.; Pires, P.S.; Lobato, F.C.F.

    2009-01-01

    Clostridium perfringens is a normal inhabitant of the intestinal tract of chickens as well as a potential pathogen that causes necrotic enteritis and colangio hepatitis. The minimum inhibitory concentration (MIC) of seven different compounds used for therapy, growth promotion or prevention of coccidiosis was determined by agar dilution method for 55 C. perfringens strains isolated from the intestines of broiler chickens. All strains showed high susceptibility to penicillin, avilamycin, monensin and narasin. Only 7.3% of the strains showed an intermediated sensitivity to lincomycin, and 49 (89.1%) were considered susceptible. For tetracycline and bacitracin, 41.8% and 47.3% of strains, respectively, were considered resistant. PMID:24031355

  9. Genomic analyses of Clostridium perfringens isolates from five toxinotypes.

    PubMed

    Hassan, Karl A; Elbourne, Liam D H; Tetu, Sasha G; Melville, Stephen B; Rood, Julian I; Paulsen, Ian T

    2015-05-01

    Clostridium perfringens can be isolated from a range of environments, including soil, marine and fresh water sediments, and the gastrointestinal tracts of animals and humans. Some C. perfringens strains have attractive industrial applications, e.g., in the degradation of waste products or the production of useful chemicals. However, C. perfringens has been most studied as the causative agent of a range of enteric and soft tissue infections of varying severities in humans and animals. Host preference and disease type in C. perfringens are intimately linked to the production of key extracellular toxins and on this basis toxigenic C. perfringens strains have been classified into five toxinotypes (A-E). To date, twelve genome sequences have been generated for a diverse collection of C. perfringens isolates, including strains associated with human and animal infections, a human commensal strain, and a strain with potential industrial utility. Most of the sequenced strains are classified as toxinotype A. However, genome sequences of representative strains from each of the other four toxinotypes have also been determined. Analysis of this collection of sequences has highlighted a lack of features differentiating toxinotype A strains from the other isolates, indicating that the primary defining characteristic of toxinotype A strains is their lack of key plasmid-encoded extracellular toxin genes associated with toxinotype B to E strains. The representative B-E strains sequenced to date each harbour many unique genes. Additional genome sequences are needed to determine if these genes are characteristic of their respective toxinotypes. PMID:25445567

  10. Suspected neurotoxicity due to Clostridium perfringens type B in a tiger (Panthera tigris).

    PubMed

    Zeira, Offer; Briola, Chiara; Konar, Martin; Dumas, Maria Pia; Wrzosek, Marcin Adam; Papa, Valentina

    2012-09-01

    A 4-yr-old tiger (Panthera tigris) was referred with acute onset of severe abnormal consciousness. Neurological evaluation showed normal palpebral and corneal reflexes, normal pupil diameter with normal direct and consensual papillary light reflex, and absent menace response bilaterally. Diffuse forebrain lesion or focal lesion affecting the ascending reticular activating system was suspected. Complete blood examination and cerebrospinal fluid analysis were normal. Magnetic resonance imaging of the brain showed an empty sella as the only result. Clostridium perfringens 10(4) to 10(7) colony-forming units/g were detected in fecal flora samples. Multiplex polymerase chain reaction assay identified serotype B counts with production of epsilon toxin. This toxin specifically accumulates in the central nervous system, where it causes acute neurological signs in humans, domestic animals, and wildlife. In this communication, the acute onset of neurological signs without evidence of trauma, vascular, metabolic, or inflammatory diseases may be caused by neurotoxicity due to C. perfringens. PMID:23082539

  11. An observation of Clostridium perfringens in Greater Sage-Grouse.

    PubMed

    Hagen, Christian A; Bildfell, Robert J

    2007-07-01

    Mortality due to infectious diseases is seldom reported in the Greater Sage-Grouse (Centrocercus urophasianus). A case of necrotic enteritis associated with Clostridium perfringens type A is described in a free-ranging adult male sage-grouse in eastern Oregon. Clostridial enteritis is known to cause outbreaks of mortality in various domestic and wild birds, and should be considered as a potential cause of mortality in sage-grouse populations. PMID:17699098

  12. Genetic diversity of Clostridium perfringens type A isolates from animals, food poisoning outbreaks and sludge

    Microsoft Academic Search

    Anders Johansson; Anna Aspan; Elisabeth Bagge; Viveca Båverud; Björn E Engström; Karl-Erik Johansson

    2006-01-01

    BACKGROUND: Clostridium perfringens, a serious pathogen, causes enteric diseases in domestic animals and food poisoning in humans. The epidemiological relationship between C. perfringens isolates from the same source has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE). In this study the genetic diversity of C. perfringens isolated from various animals, from food poisoning outbreaks and from sludge was investigated.

  13. Molecular methods for the analysis of Clostridium perfringens relevant to food hygiene

    Microsoft Academic Search

    Barbara Schalch; Brigitte Sperner; Hartmut Eisgruber; Andreas Stolle

    1999-01-01

    Clostridium perfringens continues to be a common cause of food-borne disease [1,2]. It produces an enterotoxin (CPE) which is released upon lysis of the vegetative cell during sporulation in the intestinal tract. Catering premises with insufficient cooling and reheating devices often seem to be the cause of outbreaks of C. perfringens food poisoning. Typing of C. perfringens is of great

  14. Multilocus Sequence Typing Analysis of Clostridium perfringens Isolates from Necrotic Enteritis Outbreaks in Broiler Chicken Populations

    Microsoft Academic Search

    G. Chalmers; H. L. Bruce; D. B. Hunter; V. R. Parreira; R. R. Kulkarni; Y.-F. Jiang; J. F. Prescott; P. Boerlin

    2008-01-01

    Clostridium perfringens is an important pathogen of animals and humans and is the causative agent of necrotic enteritis (NE) in poultry. This study focuses on the typing of intestinal C. perfringens isolates (n 61) from outbreaks of NE collected from several areas of Southern Ontario, using a recently developed multilocus sequence typing (MLST) technique. For comparison, C. perfringens isolates from

  15. Fatal case of Clostridium perfringens enteritis and bacteraemia in South Africa.

    PubMed

    Smith, Anthony M; Thomas, Juno; Mostert, Pieter J H

    2011-05-01

    Clostridium perfringens is an important anaerobic pathogen causing foodborne and non-foodborne gastrointestinal diseases in humans and animals. This pathogen is also the more common Clostridium species associated with bacteraemia. We report on a fatal case of C. perfringens infection in an adult with type 2 diabetes. PMID:21628819

  16. Hydrolyzable and condensed tannins resistance in Clostridium perfringens.

    PubMed

    Redondo, L M; Dominguez, J E; Rabinovitz, B C; Redondo, E A; Fernández Miyakawa, M E

    2015-08-01

    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 (antimicrobial growth promoters), 9 therapeutic antimicrobials and 2 tannin based extracts was determined for 30 C. perfringens strains isolated from poultry and cattle. Two susceptible strains were selected and cultured in presence of sub-inhibitory concentrations of tannins and AGPs for resistant sub-populations selection. Tannin resistance of C. perfringens isolates from both animal species revealed no statistically significant differences in MICs (minimum inhibitory concentration). Poultry isolates showed higher MICs to several AGPs compared with cattle isolates. All isolates were susceptible to the therapeutic antimicrobials tested, but avian isolates showed a significantly lower susceptibility to these antimicrobials which was highly correlated with an increased resistance to bacitracin and others AGPs. In-vitro selection of resistant clones suggests that C. perfringens was unable to develop resistance against tannins at least compared to AGPs like bacitracin and avilamycin. Avian origin strains, which were previously exposed to antibiotics showed higher resistance, compared to cattle origin strains. These results suggest that the evolution of resistance against tannins in C. perfringens would be more difficult and slower than to the determined AGPs. PMID:26037239

  17. Identification and cloning of two immunogenic Clostridium perfringens proteins, elongation factor Tu and pyruvate:ferredoxin oxidoreductase of C. perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium-related poultry diseases such as necrotic enteritis (NE) and gangrenous dermatitis (GD) cause substantial economic losses on a global scale. Two antigenic Clostridium perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO), were identified by react...

  18. Hospital outbreak of clostridium perfringens food-poisoning.

    PubMed

    Thomas, M; Noah, N D

    1977-05-14

    An outbreak of Clostridium perfringens (C.welchii) food-poisoning affected a third of the patients in a large hospital, and one frail patient died. C.perfringens type A, serotype 1, was isolated from 46 (61 per cent) of 76 patients examined and from food, and a new serotype (61) was isolated from 16. The attack-rate among patients who ate a minced-ham dish was 78 per cent. The cooking and storage of this mince was faulty: cuts of meat were much too large, they were kept at room temperature too long before refrigeration, and after cooking they were put into mincers used also for raw meat. C.perfringens type A, serotype 1, was isolated from meat scraps in a mincer. Final reheating was inadequate to destroy even vegetative bacteria, and multiplication may have occurred during slow distribution to the wards. Outbreaks of C.perfringens fool-poisoning are common in hospitals, and some underlying problems are discussed. A plea is made for the Food Hygiene Regulations to apply to National Health Service premises and for simple but effective reforms in institutional catering management. PMID:67498

  19. Reduction of bilirubin ditaurate by the intestinal bacterium Clostridium perfringens.

    PubMed

    Koní?ková, Renata; Jirásková, Alena; Zelenka, Jaroslav; Lešetický, Ladislav; Štícha, Martin; Vítek, Libor

    2012-01-01

    Bilirubin is degraded in the human gut by microflora into urobilinoids. In our study we investigated whether the bilirubin-reducing strain of Clostridium perfringens can reduce bilirubin ditaurate (BDT), a bile pigment of some lower vertebrates, without hydrolysis of the taurine moiety. C. perfringes was incubated under anaerobic conditions with BDT; reduction products were quantified by spectrophotometry and separated by TLC. Based on Rf values of BDT reduction products and synthetic urobilinogen ditaurate, three novel taurine-conjugated urobilinoids were identified. It is likely that bilirubin-reducing enzyme(s) serve for the effective disposal of electrons produced by fermentolytic processes in these anaerobic bacteria. PMID:22540115

  20. Determination of the Clostridium perfringens-binding site on fibronectin.

    PubMed

    Katayama, Seiichi; Tagomori, Mika; Morita, Naomi; Yamasaki, Tsutomu; Nariya, Hirofumi; Okada, Mariko; Watanabe, Mariko; Hitsumoto, Yasuo

    2015-08-01

    The extracellular matrix protein fibronectin (Fn) is known to bind to the surface of Clostridium perfringens cells. Fn is a disulfide-linked homodimer protein, with each Fn polypeptide consisting of three types of repeating modules: 12 type I, 2 type II, and 15-17 type III modules. To determine the epitope on Fn recognized by C. perfringens cells, anti-Fn monoclonal antibodies (mAbs) and various Fn fragments (III2-10, rIII2-4, rIII5-7, rIII8, rIII9, rIII10) were employed. Although two C. perfringens-derived Fn-binding proteins, FbpA and FbpB, have been reported, they appear not to be the bacterium's surface Fn receptor. Moreover, both FbpA and FbpB were found to bind to C. perfringens cells. To avoid confusion, a mutant C. perfringens lacking both the fbpA and fbpB genes (MW5) was prepared using an in-frame deletion system. MW5 cells bound Fn on their surface, suggesting the presence of a putative Fn receptor(s) on C. perfringens cells. Of several anti-Fn mAbs, both HB39 and MO inhibited the binding of Fn to MW5 cells. HB39 reacted strongly with III2-10 and rIII9, and weakly with rIII2-4, rIII10 and rIII5-7 in Western blotting analysis. Binding of HB39 to Fn was inhibited in the presence of either rIII9 or rIII10, but not in the presence of rIII2-4, rIII5-7, or rIII8. Binding of Fn to MW5 cells was strongly inhibited by both III2-10 and rIII9, marginally inhibited by rIII2-4, but not affected by rIII5-7, rIII8, or rIII10. Significant binding of MW5 cells to immobilized rIII9 and rIII10 as well as immobilized III2-10 was observed. The region of Fn recognized by C. perfringens was thus mapped to the region encompassed by III9 and III10. PMID:25433150

  1. Clostridium perfringens Type E Virulence Traits Involved in Gut Colonization

    PubMed Central

    Redondo, Leandro M.; Carrasco, Juan M. Díaz; Redondo, Enzo A.; Delgado, Fernando; Miyakawa, Mariano E. Fernández

    2015-01-01

    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 A cells. In-vitro and in-vivo models suggest that toxinotype E would be particularly adapted to exploit the changes induced by iota toxin in the surface of epithelial cells. In addition, type E strains produce metabolites that affected the growth of potential intra-specific competitors. These results suggest that the alteration of the enterocyte morphology induced by iota toxin concomitantly with the specific increase of type E cell adhesion and the strong intra-specific growth inhibition of other strains could be competitive traits inherent to type E isolates that improve its fitness within the bovine gut environment. PMID:25799452

  2. Cloning and expression of the phospholipase C gene from Clostridium perfringens and Clostridium bifermentans.

    PubMed Central

    Tso, J Y; Siebel, C

    1989-01-01

    The phospholipase C gene from Clostridium perfringens was isolated, and its sequence was determined. It was found that the structural gene codes for a protein of 399 amino acid residues. The NH2-terminal residues have the typical features of a signal peptide and are probably cleaved after secretion. Escherichia coli cells harboring the phospholipase C gene-containing plasmid expressed high levels of this protein in the periplasmic space. Phospholipase C purified from E. coli transformants was enzymatically active, hemolytic to erythrocytes, and toxic to animals when injected intravenously. The phospholipase C gene from a related organism, Clostridium bifermentans, was also isolated. The two phospholipase C genes were found to be 64% homologous in coding sequence. The C. bifermentans protein, however, was 50-fold less active enzymatically than the C. perfringens enzyme. Images PMID:2536356

  3. 9 CFR 113.112 - Clostridium Perfringens Type D Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Inactivated Bacterial Products § 113.112 Clostridium Perfringens Type D Toxoid and...

  4. 9 CFR 113.111 - Clostridium Perfringens Type C Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Inactivated Bacterial Products § 113.111 Clostridium Perfringens Type C Toxoid and...

  5. Crystallization and preliminary X-ray diffraction studies of delta-toxin from Clostridium perfringens.

    PubMed

    Huyet, Jessica; Gilbert, Maryse; Popoff, Michel R; Basak, Ajit

    2011-03-01

    Clostridium perfringens is a Gram-positive anaerobic bacterium that is responsible for a wide range of diseases in humans and both wild and domesticated animals, including birds. C. perfringens is notable for its ability to produce a plethora of toxins, e.g. phospholipases C (alpha-toxin), pore-forming toxins (epsilon-toxin, beta-toxin and enterotoxin) and binary toxins (iota-toxin). Based on alpha-, beta-, epsilon- and iota-toxin production, the bacterium is classified into five different toxinotypes (A-E). Delta-toxin, which is a 32.6?kDa protein with 290 amino acids, is one of three haemolysins released by type C and possibly by type B strains of C. perfringens. This toxin is immunogenic and lytic to erythrocytes from the even-toed ungulates sheep, goats and pigs, and is cytotoxic to other cell types such as rabbit macrophages, human monocytes and blood platelets from goats, rabbits, guinea pigs and humans. The recombinant delta-toxin has been cloned, expressed, purified and crystallized in two different crystal forms by the hanging-drop vapour-diffusion method. Of these two different crystal forms, only the form II crystal diffracted to atomic resolution (dmin=2.4?Å), while the form I crystal diffracted to only 15?Å resolution. The form II crystals belonged to space group P2(1)2(1)2, with one molecule in the crystallographic asymmetric unit and unit-cell parameters a=49.66, b=58.48, c=112.93?Å. PMID:21393845

  6. Association between gizzard lesions and increased caecal Clostridium perfringens counts in broiler chickens

    Microsoft Academic Search

    Margarita Novoa-Garrido; Stig Larsen; Magne Kaldhusdal

    2006-01-01

    The aim of this study was to investigate the relationship between mucosal gizzard lesions and caecal Clostridium perfringens counts. Gross pathological changes in the gizzard and small intestine, and caecal C. perfringens counts from 1217 meat-type chickens were recorded during the course of six experiments and were statistically analysed. C. perfringens counts increased significantly (P?

  7. Potential for growth of Clostridium perfringens from spores in pork scrapple during cooling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We conducted stabilization studies to determine the ability of Clostridium perfringens spores to germinate and grow during exponential cooling of a commercial formulation of pork scrapple. Scrapple was inoculated with a mixture of three strains of C. perfringens spores (NTCC 8238, NCTC 8239, and AT...

  8. Inhibition of Clostridium perfringens sporulation by Bacteroides fragilis and short-chain fatty acids

    Microsoft Academic Search

    Dorothy M. Wrigley

    2004-01-01

    The effect a common fecal organism, Bacteroides fragilis, has on the sporulation of Clostridium perfringens, an organism linked to some cases of antibiotic associated diarrhea, was examined. Established B. fragilis cultures significantly decreased the number of heat resistant spores formed by C. perfringens ATCC 12915 and increased the number of vegetative cells. To determine if short-chain fatty acids (SCFA), fermentation

  9. THE GENOME SEQUENCE OF BACTERIOPHAGE CpV1 LYTIC FOR CLOSTRIDIUM PERFRINGENS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Application of bacteriophages and their lytic enzymes to control Clostri-dium perfringens is one potential approach to reduce the pathogen on poultry farms and in poultry-processing facilities. We have established a collection of 30 bacteriophages lytic for C. perfringens. These were isolated from s...

  10. Clostridium perfringens Spore Germination: Characterization of Germinants and Their Receptors?

    PubMed Central

    Paredes-Sabja, Daniel; Torres, J. Antonio; Setlow, Peter; Sarker, Mahfuzur R.

    2008-01-01

    Clostridium perfringens food poisoning is caused by type A isolates carrying a chromosomal enterotoxin (cpe) gene (C-cpe), while C. perfringens-associated non-food-borne gastrointestinal (GI) diseases are caused by isolates carrying a plasmid-borne cpe gene (P-cpe). C. perfringens spores are thought to be the important infectious cell morphotype, and after inoculation into a suitable host, these spores must germinate and return to active growth to cause GI disease. We have found differences in the germination of spores of C-cpe and P-cpe isolates in that (i) while a mixture of l-asparagine and KCl was a good germinant for spores of C-cpe and P-cpe isolates, KCl and, to a lesser extent, l-asparagine triggered spore germination in C-cpe isolates only; and (ii) l-alanine or l-valine induced significant germination of spores of P-cpe but not C-cpe isolates. Spores of a gerK mutant of a C-cpe isolate in which two of the proteins of a spore nutrient germinant receptor were absent germinated slower than wild-type spores with KCl, did not germinate with l-asparagine, and germinated poorly compared to wild-type spores with the nonnutrient germinants dodecylamine and a 1:1 chelate of Ca2+ and dipicolinic acid. In contrast, spores of a gerAA mutant of a C-cpe isolate that lacked another component of a nutrient germinant receptor germinated at the same rate as that of wild-type spores with high concentrations of KCl, although they germinated slightly slower with a lower KCl concentration, suggesting an auxiliary role for GerAA in C. perfringens spore germination. In sum, this study identified nutrient germinants for spores of both C-cpe and P-cpe isolates of C. perfringens and provided evidence that proteins encoded by the gerK operon are required for both nutrient-induced and non-nutrient-induced spore germination. PMID:18083820

  11. Biological Characteristics of Clostridium perfringens Type A Enterotoxin

    PubMed Central

    Stark, Raymond L.; Duncan, Charles L.

    1971-01-01

    An enterotoxin with the ability to induce fluid accumulation in rabbit ileal loops, erythema in the skin of guinea pigs, and lethality in mice appears in cell extracts (CE) and culture filtrates (CF) of sporulating cells of some Clostridium perfringens type A strains. All activities in CE and CF were eluted simultaneously from a Sephadex G-200 column. Different elution patterns were obtained for these activities present in CE and CF. Rabbit immune serum against CF and the active CE fractions eliminated the three biological activities in CE and CF. These activities present in CF and CE were not eliminated by any of the known antitoxins present in diagnostic serum against C. perfringens types A, B, C, D, and E. Immunodiffusion studies with immune serum against active CE fractions and CF indicated a precipitin line of identity between CF and CE of NCTC 8798 and other enterotoxin-positive strains but not enterotoxin-negative strains. Disc electrophoresis of active G-200 fractions on 7.0% polyacrylamide gels revealed a single area containing erythemal activity and mouse lethality. Immunodiffusion with acrylamide gels, containing crude fractionated enterotoxin, and immune serum against partially purified enterotoxin revealed a single precipitin band in the same area as the biological activities. Immunoelectrophoresis of CE of enterotoxin-positive and enterotoxin-negative strains also showed one precipitin band which occurred only with enterotoxin-positive strains. These findings suggest that one component is responsible for the biological activities attributed to the enterotoxin. Images PMID:4343403

  12. Cold shock lethality and injury in Clostridium perfringens.

    PubMed

    Traci, P A; Duncan, C L

    1974-11-01

    Several observations have been made in regard to cold shock lethality of Clostridium perfringens: (i) loss of viability was not consequence of exposure of the cells to air; (ii) stationary-phase cells were much more resistant to cold shock at 4 C than exponential-phase cells; (iii) at 4 C 96% of an initial population of exponential-phase cells was killed upon cold shock and 95% of the remaining population was killed within 90 min of continued exposure at 4 C; (iv) the minimal temperature differential for detectable cold shock lethality was between 17 and 23 C, and the maximum beyond which lethality was not appreciably increased was between 28 and 33 C. Up to 75% of viable cold-shocked cells were injured, as demonstrated by cold shocking late exponential-phase cells at 10 C and using differential plating procedure for recovery. Repair of injury was temperature dependent, and occurred in a complex medium and 0.1% peptone but not water. Nalidixic acid, chloramphenicol, and rifampin did not inhibit repair of injury. PMID:4374121

  13. Clostridium perfringens type A enteritis in blue and yellow macaw (Ara ararauna).

    PubMed

    Guimarães, Marta Brito; Torres, Luciana Neves; Mesquita, Ramon Gomes; Ampuero, Fernanda; Cunha, Marcos Paulo Vieira; Ferreira, Thais Sebastiana Porfida; Ferreira, Antonio José Piantino; Catão-Dias, José Luiz; Moreno, Andrea Micke; Knöbl, Terezinha

    2014-12-01

    This study describes an outbreak of necrotic enteritis caused by Clostridium perfringens type A in captive macaws (Ara ararauna). Two psittacine birds presented a history of prostration and died 18 hr after manifestation of clinical signs. The necropsy findings and histopathologic lesions were indicative of necrotic enteritis. Microbiologic assays resulted in the growth of large gram-positive bacilli that were identified as C. perfringens. PCR was used to identify clostridium toxinotypes and confirmed the identification of isolated strains as C pefringens type A, positive to gene codifying beta 2 toxin. The infection source and predisposing factors could not be ascertained. PMID:25619013

  14. Comparison of virulence plasmids among Clostridium perfringens type E isolates.

    PubMed

    Li, Jihong; Miyamoto, Kazuaki; McClane, Bruce A

    2007-04-01

    Clostridium perfringens type E isolates produce iota-toxin, which is encoded by iap and ibp genes. Using Southern blot analyses, the current study identified iap/ibp plasmids of approximately 97 or approximately 135 kb among eight type E isolates. For most of these isolates, their iap/ibp plasmid also encoded urease and lambda-toxin. However, the beta2-toxin gene, if present, was on a different plasmid from the iap/ibp plasmid. For all isolates, the iap/ibp plasmid carried a tcp locus, strongly suggesting that these plasmids are conjugative. Overlapping PCR analyses demonstrated some similarity between the iap/ibp plasmids and enterotoxin-encoding plasmids of type A isolates. Additional PCR analyses demonstrated that the iap/ibp locus is located near dcm sequences, an apparent plasmid hot spot for toxin gene insertion, and that two IS1151-related sequences are present in the iap/ibp locus. To begin testing whether those IS1151-like sequences can mobilize iap/ibp genes, a PCR assay was performed that amplifies a product only from circular DNA forms that could represent transposition intermediates. This PCR assay detected circular forms containing iap/ibp genes and silent enterotoxin gene sequences, with or without an IS1151-like sequence. Collectively, these results suggest that a mobile genetic element carrying iap/ibp has inserted onto a tcp-carrying enterotoxin plasmid in a type A isolate, creating a progenitor iap/ibp plasmid. That plasmid then spread via conjugation to other isolates, converting them to type E. Further iap/ibp plasmid diversity occurred when either the iap/ibp genes later remobilized and inserted onto other conjugative plasmids or some iap/ibp plasmids acquired additional DNA sequences. PMID:17261608

  15. Identification and cloning of two immunogenic Clostridium perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO) of C. perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium-related diseases such as gangrenous dermatitis (GD) and necrotic enteritis (NE) are increasingly emerging as major diseases in recent years with high economic loss around the world. In this report, we characterized two immunogenic Clostridium perfringens (CP) proteins (e.g., elongation f...

  16. EVALUATION OF AN EXPERIMENTAL CHLORATE PRODUCT ON CLOSTRIDIUM PERFRINGENS IN A MIXED GUT CULTURE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium perfringens (Cp), the etiologic agent of necrotic enteritis is an economically important pathogen of the poultry industry; causing significant morbidity and mortality in commercial flocks worldwide. As pressure mounts to discontinue the use of antibiotics in the agriculture industry, it...

  17. Molecular Characterization of Podoviridae Bacteriophages Virulent for Clostridium perfringens and Comparison of Their Predicted Lytic Proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium perfringens is a Gram-positive, spore-forming anaerobic bacterium that plays a significant role in human food-borne disease as well as non-food-borne human, animal and poultry diseases. There has been a resurgent interest in the use of bacteriophages or their gene products to control ba...

  18. BACTERIOPHAGES OF THE FAMILY SIPHOVIRIDAE CONTAIN AMIDASE ENZYMES THAT LYSE CLOSTRIDIUM PERFRINGENS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In chickens Clostridium perfringens (Cp) is the etiologic agent of necrotic enteritis and causes gas gangrene along with being the third leading cause of bacterial food-borne gastroenteritis in humans. While the disease in poultry can be controlled by antibiotics, there is increasing pressure to ban...

  19. Bacteriophages of the family siphoviridae contain amidase enzymes that lyse Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    *Agtech-Danisco, current address In chickens Clostridium perfringens (Cp) is the etiologic agent of necrotic enteritis and causes gas gangrene along with being the third leading cause of bacterial food-borne gastroenteritis in humans. While the disease in poultry can be controlled by antibiotics, th...

  20. In-vitro antimicrobial susceptibility of Clostridium perfringens from commercial turkey and broiler chicken origin

    Microsoft Academic Search

    K. L. Watkins; T. R. Shryock; R. N. Dearth; Y. M. Saif

    1997-01-01

    The minimum inhibitory concentrations (MIC) of eight antibiotics and two anticoccidial agents were determined for Clostridium perfringens strains isolated from 26 commercial broiler farms and 22 commercial turkey farms. Isolates were obtained from the intestines of birds on the farm or at the processing plant using standard culture and identification techniques. The microbroth dilution test was used to determine the

  1. Clonal relationships among Clostridium perfringens of porcine origin as determined by multilocus sequence typing

    Microsoft Academic Search

    B. Helen Jost; Hien T. Trinh; J. Glenn Songer

    2006-01-01

    Clostridium perfringens is ubiquitous in the environment and the intestinal tracts of most mammals, but this organism also causes gas gangrene and enteritis in human and animal hosts. While expression of specific toxins correlates with specific disease in certain hosts, the other factors involved in commensalism and host pathogenesis have not been clearly identified. A multilocus sequence typing (MLST) scheme

  2. CLOSTRIDIUM PERFRINGENS: STATUS OF A FOOD-BORNE SPORE-FORMER

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium perfringens is responsible for the third most common cause of food-borne illness in the U.S. today, resulting in an estimated 0.25 million cases annually and an associated economic loss of 12.5 billion dollars. The increased production of minimally-processed, extended shelf-life, refrig...

  3. Detection of Clostridium perfringens type C in pig herds following disease outbreak and subsequent vaccination.

    PubMed

    Schäfer, K; Wyder, M; Gobeli, S; Candi, A; Doherr, M G; Zehnder, B; Zimmermann, W; Posthaus, H

    2012-11-17

    Immunisation of sows using Clostridium perfringens type C toxoid vaccines is recommended to prevent necrotising enteritis (NE) on pig breeding farms. Absence of disease, however, oftentimes leads to the false assumption of pathogens being eradicated. The prevalence of C perfringens type C was determined by PCR in faecal samples of piglets and sows in three Swiss pig breeding farms two to four years after implementation of a vaccination programme following disease outbreaks. C perfringens type C could still be detected several years after an outbreak despite absence of NE. In-herd prevalence of the pathogens varied significantly between the farms and was also lower compared with a farm which experienced a recent outbreak. In conclusion, C perfringens type C can be detected on once-affected farms, even in the absence of NE for several years. PMID:23100304

  4. Clostridium perfringens bacteriophages FCP39O and FCP26F: genomic organization and proteomic analysis of the virions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Initial screening for bacteriophages lytic for Clostridium perfringens was performed utilizing filtered samples obtained from poultry (intestinal material), soil, sewage and poultry processing drainage water. Lytic phage preparations were initially characterized by transmission electron microscopy ...

  5. Genotyping of Enterotoxigenic Clostridium perfringens Fecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

    Microsoft Academic Search

    SHAUNA G. SPARKS; ROBERT J. CARMAN; MAHFUZUR R. SARKER

    2001-01-01

    Clostridium perfringens type A isolates producing enterotoxin (CPE) are an important cause of food poisoning and non-food-borne human gastrointestinal (GI) diseases, including antibiotic-associated diarrhea (AAD). Recent studies suggest that C. perfringens type A food poisoning is caused by C. perfringens isolates carrying a chromosomal cpe gene, while CPE-associated non-food-borne GI diseases, such as AAD, are caused by plasmid cpe isolates.

  6. Roles of DacB and Spm Proteins in Clostridium perfringens Spore Resistance to Moist Heat, Chemicals, and UV Radiation

    Microsoft Academic Search

    Daniel Paredes-Sabja; Nahid Sarker; Barbara Setlow; Peter Setlow; Mahfuzur R. Sarker

    Clostridium perfringens food poisoning is caused mainly by enterotoxigenic type A isolates that typically possess high spore heat resistance. Previous studies have shown that \\/-type small, acid-soluble proteins (SASP) play a major role in the resistance of Bacillus subtilis and C. perfringens spores to moist heat, UV radiation, and some chemicals. Additional major factors in B. subtilis spore resistance are

  7. Predictive model for growth of Clostridium perfringens at temperatures applicable to cooling of cooked uncured beef and chicken

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this investigation was to develop and validate a model for predicting the relative growth of Clostridium perfringens from spore inocula in uncured chicken and beef meat during cooling. Isothermal growth curves of C. perfringens at various temperatures from 10-48.9C were evaluated, ...

  8. Clostridium perfringens: A review of enteric diseases in dogs, cats and wild animals.

    PubMed

    Silva, Rodrigo Otávio Silveira; Lobato, Francisco Carlos Faria

    2015-06-01

    Clostridium perfringens is a gram-positive anaerobic bacillus that is commonly part of the microbiota of humans and animals. It is considered a common enteric pathogen, but the pathogenesis and the predisposing factors of the disease commonly differ between host species. Thus, specific research is necessary to understand the role of this pathogen, how to diagnose it, and which control measures are applicable. The aim of this paper is to review the current knowledge of C. perfringens infections in dogs, cats and wild animals. PMID:25644183

  9. Residues involved in the pore-forming activity of the Clostridium perfringens iota toxin.

    PubMed

    Knapp, Oliver; Maier, Elke; Waltenberger, Eva; Mazuet, Christelle; Benz, Roland; Popoff, Michel R

    2015-02-01

    Clostridium perfringens iota toxin is a binary toxin that is organized into enzyme (Ia) and binding (Ib) components. Ib forms channels in lipid bilayers and mediates the transport of Ia into the target cells. Here we show that Ib residues 334-359 contain a conserved pattern of alternating hydrophobic and hydrophilic residues forming two amphipathic ?-strands involved in membrane insertion and channel formation. This stretch of amino acids shows remarkable structural and functional analogies with the ?-pore-forming domain of C. perfringens epsilon toxin. Several mutations within the two amphipathic ?-strands affected pore formation, single-channel conductance and ion selectivity (S339E-S341E, Q345H N346E) confirming their involvement in channel formation. F454 of Ib corresponds to the ?-clamp F427 of anthrax protective antigen and F428 of C2II binary toxins. The mutation F454A resulted in a loss of cytotoxicity and strong increase in single-channel conductance (500 pS as compared with 85?pS in 1 M KCl) with a slight decrease in cation selectivity, indicating that the ?-clamp is highly conserved and crucial for binary toxin activity. In contrast, the mutants Q367D, N430D, L443E had no or only minor effects on Ib properties, while T360I, T360A and T360W caused a dramatic effect on ion selectivity and single-channel conductance, indicating gross disturbance of the oligomer structure. This suggests that, at least in the iota toxin family, T360 has a structural role in the pore organization. Moreover, introduction of charged residues within the channel (S339E-S341E) or in the vestibule (Q367D, N430D and L443E) had virtually no effect on chloroquine or Ia binding, whereas F454A, T360I, T360A and T360W strongly decreased the chloroquine and Ia affinity to Ib. These results support that distinct residues within the vestibule interact with chloroquine and Ia or are responsible for channel structure, while the channel lining amino acids play a less important role. PMID:25266274

  10. Complete genome sequence of the podoviral bacteriophage ?CP24R, which is virulent for Clostridium perfringens.

    PubMed

    Morales, Cesar A; Oakley, Brian B; Garrish, Johnna K; Siragusa, Gregory R; Ard, Mary B; Seal, Bruce S

    2012-04-01

    Bacteriophage ?CP24R was isolated from raw sewage from a waste treatment plant, and lytic activity was observed against a type A Clostridium perfringens isolate. Electron microscopy revealed a small virion (44-nm-diameter icosahedral capsid) with a short, non-contractile tail, indicative of a member of the family Podoviridae. The phage had a linear, double-stranded DNA genome of 18,919 base pairs (bp) with 41 bp inverted terminal repeats and a type B DNA polymerase, which are characteristics of members of the subfamily Picovirinae. Out of 22 predicted genes in the genome, ten had significant sequence similarity to proteins of known function. Three distinct genes with lytic domains were identified, including a zinc carboxypeptidase domain that has not been previously reported in viruses. The ?CP24R genome described herein is only the second Clostridium perfringens podovirus genome reported to date. PMID:22218967

  11. The induction of toxin neutralizing antibodies to Clostridium perfringens types C and D toxins in horses

    E-print Network

    Brooks, Frances Lynn

    1983-01-01

    THE INDUCTION OF TOXIN NEUTRALIZING ANTIBODIES TO CLOSTRIDIUM PERFRINGENS TYPES C AND D TOXINS IN HORSES A Thes1s by FRANCES LYNN BROOKS Submitted to the Graduate College of Texas A&M University in partial fulfillment of the requirements... and content by: Russe B. Simpson~~ (Chairman of Committee) (21 ) /'' James E. Grimes (Nember) John M. Bowen (Nember) Ian R. Tizard (Head of Department) December 1983 ABSTRACT The Induction of Toxin Neutraliz1ng Ant1bodies to Clostridium Oeeff ti...

  12. The induction of toxin neutralizing antibodies to Clostridium perfringens types C and D toxins in horses 

    E-print Network

    Brooks, Frances Lynn

    1983-01-01

    THE INDUCTION OF TOXIN NEUTRALIZING ANTIBODIES TO CLOSTRIDIUM PERFRINGENS TYPES C AND D TOXINS IN HORSES A Thes1s by FRANCES LYNN BROOKS Submitted to the Graduate College of Texas A&M University in partial fulfillment of the requirements... and content by: Russe B. Simpson~~ (Chairman of Committee) (21 ) /'' James E. Grimes (Nember) John M. Bowen (Nember) Ian R. Tizard (Head of Department) December 1983 ABSTRACT The Induction of Toxin Neutraliz1ng Ant1bodies to Clostridium Oeeff ti...

  13. Spoilage of an acid food product by Clostridium perfringens, C. barati and C. butyricum.

    PubMed

    de Jong, J

    1989-05-01

    Spoilage of canned pasteurized brined mung bean sprouts, acidified with citric acid to pH 4.0-4.5, was found to be caused by acid tolerant Clostridium spp. including the species barati, perfringens and butyricum. The pH limit for growth in the brine used were estimated 3.7, 3.7 and 4.0 respectively. Some of the isolated C. perfringens strains produced enterotoxins in sporulation media. The spores of the isolated anaerobes appeared to originate from mung beans, but C. barati and C. perfringens strains freshly isolated from dry beans, were unable to grow in acidified brine. During germination and sprouting of mung beans, the oxygen concentration decreased, while carbon dioxide concentration increased considerably, due to respiration of the sprouts and actively growing Enterobacteriaceae and lactobacilli. It was assumed that this allowed C. barati and C. perfringens strains to grow and acquire the observed unusual acid tolerance. After increasing aerobicity during sprouting, no growth of Clostridium spp. was observed, substantiating the assumption. PMID:2561952

  14. Aqueous two-phase systems extraction of ?-toxin from Clostridium perfringens type A

    Microsoft Academic Search

    Maria Taciana Holanda Cavalcanti; Tatiana Souza Porto; Benicio de Barros Neto; José Luiz Lima-Filho; Ana Lúcia Figueiredo Porto; Adalberto Pessoa

    2006-01-01

    Two sequential half-fraction designs were applied to studying the ?-toxin partition produced by Clostridium perfringens type A in aqueous two phase systems (ATPS), as a function of four factors: PEG molar mass and concentration, phosphate concentration and pH. The highest purification factor, yield and partition coefficient results were obtained with PEG 8000 (15%, w\\/w), phosphate at 20% (w\\/w) and pH

  15. Non-classical azoreductase secretion in Clostridium perfringens in response to sulfonated azo dye exposure.

    PubMed

    Morrison, Jessica M; John, Gilbert H

    2015-08-01

    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. perfringens involving the effect of azo dye exposure was completed to answer this question. Through exposure studies, azo dyes were found to cause cytoplasmic protein release, including AzoC, from C. perfringens in dividing and non-dividing cells. Sulfonation (negative charge) of azo dyes proved to be the key to facilitating protein release of AzoC and was found to be azo-dye-concentration-dependent. Additionally, AzoC was found to localize to the Gram-positive periplasmic region. Using a ?azoC knockout mutant, the presence of additional azoreductases in C. perfringens was suggested. These results support the notion that the azoreduction of these dyes may occur extracellularly for the commensal C. perfringens in the intestine. PMID:25881497

  16. Membrane vesicles of Clostridium perfringens Type A strains induce innate and adaptive immunity

    PubMed Central

    Jiang, Yanlong; Kong, Qingke; Roland, Kenneth L.; Curtiss, Roy

    2014-01-01

    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 identified by LC-MS/MS. C. perfringens MVs contained DNA components such as 16S ribosomal RNA gene (16S rRNA), alpha-toxin gene (plc) and the perfringolysin O gene (pfoA) demonstrated by PCR. We also identified a total of 431 proteins in vesicles by 1-D gel separation and LC-MS/MS analysis. In vitro studies demonstrated that vesicles could be internalized into murine macrophage RAW264.7 cells without direct cytotoxicity effects, causing release of inflammation cytokines including granulocyte colony stimulating factor (G-CSF), tumor necrosis factor-alpha (TNF-?) and interleukin-1 (IL-1), which could also be detected in mice injected with MVs through intraperitoneal (i.p.) route. Mice immunized with C. perfringens MVs produced high titer IgG, especially IgG1, antibodies against C. perfringens membrane proteins. However, this kind of antibody could not provide protection in mice following challenge, though it could slightly postpone the time of death. Our results indicate that release of MVs from C. perfringens could provide a previously unknown mechanism to induce release of inflammatory cytokines, especially TNF-?, these findings may contribute to a better understanding of the pathogenesis of C. perfringens infection. PMID:24631214

  17. Identification and cloning of two immunogenic C. perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO) of Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium related poultry diseases such as necrotic enteritis (NE) and gangrenous dermatitis (GD) cause substantial economic losses on a global scale. Two antigenic C. perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO), were identified by reaction with...

  18. Biofilm formation of Clostridium perfringens and its exposure to low-dose antimicrobials

    PubMed Central

    Charlebois, Audrey; Jacques, Mario; Archambault, Marie

    2014-01-01

    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 porcine commensal isolates. A subgroup of clinical and commensal C. perfringens isolates was randomly selected for further characterization. Biofilm was found to protect C. perfringens bacterial cells from exposure to high concentrations of tested antimicrobials. Exposure to low doses of some of these antimicrobials tended to lead to a diminution of the biofilm formed. However, a few isolates showed an increase in biofilm formation when exposed to low doses of tylosin, bacitracin, virginiamycin, and monensin. Six isolates were randomly selected for biofilm analysis using scanning laser confocal microscopy. Of those, four produced more biofilm in presence of low doses of bacitracin whereas biofilms formed without bacitracin were thinner and less elevated. An increase in the area occupied by bacteria in the biofilm following exposure to low doses of bacitracin was also observed in the majority of isolates. Morphology examination revealed flat biofilms with the exception of one isolate that demonstrated a mushroom-like biofilm. Matrix composition analysis showed the presence of proteins, beta-1,4 linked polysaccharides and extracellular DNA, but no poly-beta-1,6-N-acetyl-D-glucosamine. This study brings new information on the biofilm produced by C. perfringens and its exposure to low doses of antimicrobials. PMID:24795711

  19. New amino acid germinants for spores of the enterotoxigenic Clostridium perfringens type A isolates.

    PubMed

    Udompijitkul, Pathima; Alnoman, Maryam; Banawas, Saeed; Paredes-Sabja, Daniel; Sarker, Mahfuzur R

    2014-12-01

    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 efficiently induced germination of C. perfringens spores; L-asparagine, L-cysteine, L-serine, and L-threonine triggered germination of spores of most FP and NFB isolates; whereas, L-glutamine was a unique germinant for FP spores. For cysteine- or glutamine-induced germination, gerKC spores (spores of a gerKC mutant derivative of FP strain SM101) germinated to a significantly lower extent and released less DPA than wild type spores; however, a less defective germination phenotype was observed in gerAA or gerKB spores. The germination defects in gerKC spores were partially restored by complementing the gerKC mutant with a recombinant plasmid carrying wild-type gerKA-KC, indicating that GerKC is an essential GR protein. The gerKA, gerKC, and gerKB spores germinated significantly slower with L-serine and L-threonine than their parental strain, suggesting the requirement for these GR proteins for normal germination of C. perfringens spores. In summary, these results indicate that the polar, uncharged amino acids at pH 6.0 are effective germinants for spores of C. perfringens type A and that GerKC is the main GR protein for germination of spores of FP strain SM101 with L-cysteine, L-glutamine, and L-asparagine. PMID:25084641

  20. Inactivation strategy for Clostridium perfringens spores adhered to food contact surfaces.

    PubMed

    Udompijitkul, Pathima; Alnoman, Maryam; Paredes-Sabja, Daniel; Sarker, Mahfuzur R

    2013-06-01

    The contamination of enterotoxigenic Clostridium perfringens spores on food contact surfaces posses a serious concern to food industry due to their high resistance to various preservation methods typically applied to control foodborne pathogens. In this study, we aimed to develop an strategy to inactivate C. perfringens spores on stainless steel (SS) surfaces by inducing spore germination and killing of germinated spores with commonly used disinfectants. The mixture of l-Asparagine and KCl (AK) induced maximum spore germination for all tested C. perfringens food poisoning (FP) and non-foodborne (NFB) isolates. Incubation temperature had a major impact on C. perfringens spore germination, with 40 °C induced higher germination than room temperature (RT) (20 ± 2 °C). In spore suspension, the implementation of AK-induced germination step prior to treatment with disinfectants significantly (p < 0.05) enhanced the inactivation of spores of FP strain SM101. However, under similar conditions, no significant spore inactivation was observed with NFB strain NB16. Interestingly, while the spores of FP isolates were able to germinate with AK upon their adhesion to SS chips, no significant germination was observed with spores of NFB isolates. Consequently, the incorporation of AK-induced germination step prior to decontamination of SS chips with disinfectants significantly (p < 0.05) inactivated the spores of FP isolates. Collectively, our current results showed that triggering spore germination considerably increased sporicidal activity of the commonly used disinfectants against C. perfringens FP spores attached to SS chips. These findings should help in developing an effective strategy to inactivate C. perfringens spores adhered to food contact surfaces. PMID:23541199

  1. An investigation into the association between cpb2-encoding Clostridium perfringens type A and diarrhea in neonatal piglets.

    PubMed

    Farzan, Abdolvahab; Kircanski, Jasmina; DeLay, Josepha; Soltes, Glenn; Songer, J Glenn; Friendship, Robert; Prescott, John F

    2013-01-01

    To investigate the possible role of cpb2-positive type A Clostridium perfringens in neonatal diarrheal illness in pigs, the jejunum and colon of matched normal and diarrheic piglets from 10 farms with a history of neonatal diarrhea were examined grossly and by histopathology, and tested for C. perfringens, for C. perfringens beta2 (CPB2) toxin, as well as for Clostridium difficile toxins, Salmonella, enterotoxigenic Escherichia coli, rotavirus, transmissible gastroenteritis (TGE) virus, and coccidia. Clostridium perfringens isolates were tested using a multiplex real-time polymerase chain reaction (PCR) to determine the presence of cpa, consensus and atypical cpb2, and other virulence-associated genes. The numbers of C. perfringens in the intestinal contents were lower in diarrheic piglets (log?? 5.4 CFU/g) compared with normal piglets (log?? 6.5 CFU/g) (P < 0.05). The consensus cpb2 was present in 93% of isolates in each group, but atypical cpb2 was less common (56% healthy, 32% diarrheic piglets isolates, respectively, P < 0.05). The presence of CPB2 toxin in the intestinal contents of normal and diarrheic piglets did not differ significantly. Clostridium difficile toxins and rotavirus were each detected in 7 of the 21 (33%) diarrheic piglets. Rotavirus, C. difficile toxins, Salmonella, or enterotoxigenic E. coli were concurrently recovered in different combinations in 4 diarrheic piglets. The cause of diarrhea in 8 of the 21 (38%) piglets on 6 farms remained unknown. The etiological diagnosis of diarrhea could not be determined in any of the piglets on 2 of the farms. This study demonstrated that the number of cpb2-positive type A C. perfringens in the intestinal contents was not a useful approach for making a diagnosis of type A C. perfringens enteritis in piglets. Further work is required to confirm whether cpb2-carrying type A C. perfringens have a pathogenic role in enteric infection in neonatal swine. PMID:23814355

  2. An investigation into the association between cpb2-encoding Clostridium perfringens type A and diarrhea in neonatal piglets

    PubMed Central

    Farzan, Abdolvahab; Kircanski, Jasmina; DeLay, Josepha; Soltes, Glenn; Songer, J. Glenn; Friendship, Robert; Prescott, John F.

    2013-01-01

    To investigate the possible role of cpb2-positive type A Clostridium perfringens in neonatal diarrheal illness in pigs, the jejunum and colon of matched normal and diarrheic piglets from 10 farms with a history of neonatal diarrhea were examined grossly and by histopathology, and tested for C. perfringens, for C. perfringens beta2 (CPB2) toxin, as well as for Clostridium difficile toxins, Salmonella, enterotoxigenic Escherichia coli, rotavirus, transmissible gastroenteritis (TGE) virus, and coccidia. Clostridium perfringens isolates were tested using a multiplex real-time polymerase chain reaction (PCR) to determine the presence of cpa, consensus and atypical cpb2, and other virulence-associated genes. The numbers of C. perfringens in the intestinal contents were lower in diarrheic piglets (log10 5.4 CFU/g) compared with normal piglets (log10 6.5 CFU/g) (P < 0.05). The consensus cpb2 was present in 93% of isolates in each group, but atypical cpb2 was less common (56% healthy, 32% diarrheic piglets isolates, respectively, P < 0.05). The presence of CPB2 toxin in the intestinal contents of normal and diarrheic piglets did not differ significantly. Clostridium difficile toxins and rotavirus were each detected in 7 of the 21 (33%) diarrheic piglets. Rotavirus, C. difficile toxins, Salmonella, or enterotoxigenic E. coli were concurrently recovered in different combinations in 4 diarrheic piglets. The cause of diarrhea in 8 of the 21 (38%) piglets on 6 farms remained unknown. The etiological diagnosis of diarrhea could not be determined in any of the piglets on 2 of the farms. This study demonstrated that the number of cpb2-positive type A C. perfringens in the intestinal contents was not a useful approach for making a diagnosis of type A C. perfringens enteritis in piglets. Further work is required to confirm whether cpb2-carrying type A C. perfringens have a pathogenic role in enteric infection in neonatal swine. PMID:23814355

  3. Carbon Catabolite Repression of Type IV Pilus-Dependent Gliding Motility in the Anaerobic Pathogen Clostridium perfringens

    Microsoft Academic Search

    Marcelo Mendez; I-Hsiu Huang; Kaori Ohtani; Roberto Grau; Tohru Shimizu; Mahfuzur R. Sarker

    2008-01-01

    Clostridium perfringens is an anaerobic, gram-positive, spore-forming bacterium responsible for the produc- tion of severe histotoxic and gastrointestinal diseases in humans and animals. In silico analysis of the three available genome-sequenced C. perfringens strains (13, SM101, and ATCC13124) revealed that genes that encode flagellar proteins and genes involved in chemotaxis are absent. However, those strains exhibit type IV pilus (TFP)-dependent

  4. Use of power ultrasound to enhance the thermal inactivation of Clostridium perfringens spores in beef slurry.

    PubMed

    Evelyn; Silva, Filipa V M

    2015-08-01

    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 60min TS process (24kHz, 0.33W/g) resulted in a less than 1.5 log reduction for both C. perfringens NZRM 898 and NZRM 2621 spores. Then, the thermal inactivation first order kinetic parameters of C. perfringens spores in beef slurry were estimated for the two strains. The D105°C- and z-values were 2.5min and 10.6°C for NZRM 898 and 1.8min and 10.9°C for NZRM 2621. After, the effect of a spore heat shock followed by ultrasound on its thermal inactivation in beef slurry was investigated. This heat shock+ultrasound pretreatment was able to double the spore thermal inactivation rate in beef slurry. For example at 95°C D-value of 20.2min decreased to 9.8min, demonstrating that spore exposure to heat shock followed by ultrasonication enhanced its thermal inactivation. PMID:25912313

  5. Identification and modeling of a drug target for Clostridium perfringens SM101

    PubMed Central

    Chhabra, Gagan; Sharma, Pramila; Anant, Avishek; Deshmukh, Sachin; Kaushik, Himani; Gopal, Keshav; Srivastava, Nutan; Sharma, Neeraj; Garg, Lalit C

    2010-01-01

    In the present study, comparative genome analysis between Clostridium perfringens and the human genome was carried out to identify genes that are essential for the pathogen's survival, and non-homologous to the genes of human host, that can be used as potential drug targets. The study resulted in the identification of 426 such genes. The number of these potential drug targets thus identified is significantly lower than the genome's protein coding capacity (2558 protein coding genes). The 426 genes of C. perfringens were further analyzed for overall similarities with the essential genes of 14 different bacterial species present in Database of Essential Genes (DEG). Our results show that there are only 5 essential genes of C. perfringens that exhibit similarity with 12 species of the 14 different bacterial species present in DEG database. Of these, 1 gene was similar in 12 species and 4 genes were similar in 11 species. Thus, the study opens a new avenue for the development of potential drugs against the highly pathogenic bacterium. Further, by selecting these essential genes of C. perfringens, which are common and essential for other pathogenic microbial species, a broad spectrum anti-microbial drug can be developed. As a case study, we have built a homology model of one of the potential drug targets, ABC transporter-ATP binding protein, which can be employed for in silico docking studies by suitable inhibitors. PMID:20978600

  6. Role of GerKB in Germination and Outgrowth of Clostridium perfringens Spores? †

    PubMed Central

    Paredes-Sabja, Daniel; Setlow, Peter; Sarker, Mahfuzur R.

    2009-01-01

    Previous work indicated that Clostridium perfringens gerKA gerKC spores germinate significantly, suggesting that gerKB also has a role in C. perfringens spore germination. We now find that (i) gerKB was expressed only during sporulation, likely in the forespore; (ii) gerKB spores germinated like wild-type spores with nonnutrient germinants and with high concentrations of nutrients but more slowly with low nutrient concentrations; and (iii) gerKB spores had lower colony-forming efficiency and slower outgrowth than wild-type spores. These results suggest that GerKB plays an auxiliary role in spore germination under some conditions and is required for normal spore viability and outgrowth. PMID:19363077

  7. Ulcerative enteritis-like disease associated with Clostridium perfringens type A in bobwhite quail (Colinus virginianus).

    PubMed

    Shivaprasad, H L; Uzal, Francisco; Kokka, Randy; Fisher, Derek J; McClane, Bruce A; Songer, A Glenn

    2008-12-01

    Ulcerative enteritis-like disease due to Clostridium perfringens type A was attributed as the cause of mortality in excess of 50% in a flock of 1000, 10-to-16-wk-old bobwhite quail (Colinus virginianus). Clinical signs in these birds ranged from sudden death to listlessness, depression, watery white droppings, ruffled feathers, loss of weight, and death in a few days. Necropsy of 30 birds revealed multiple deep ulcers of the mucosa throughout the small intestine and ceca, some with perforation and subsequent coelomitis (peritonitis). The livers in some birds contained white foci of necrosis, and many birds had enlarged and congested spleens. Microscopic lesions included multifocal severe fibrinosuppurative ulcerative enteritis associated with large numbers of rod-shaped gram-positive bacteria, and necrotizing hepatitis with or without rod-shaped bacteria. Anaerobic culturing of the intestine and liver yielded pure cultures of C. perfringens. The C perfringens isolates were of genotype A and were polymerase chain reaction-positive for alpha toxin and for cpb2, the structural gene for beta2 toxin. Repeated attempts to isolate C colinum by using a specialized medium containing 8% horse plasma were not fruitful, suggesting that the enteritis and hepatitis in these birds were produced by C pefringens. Retrospective examination of records of quail submissions to the California Animal Health and Food Safety Laboratory System over 16 yr revealed at least nine quail submissions in which isolation of C. perfringens from the liver, intestine, or both was associated with ulcerative enteritis and hepatitis in quail. This is the first description of ulcerative enteritis-like disease in quail associated with C perfringens. Final conclusions await experimental reproduction of the disease. PMID:19166055

  8. Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens

    PubMed Central

    Charlebois, Audrey; Jalbert, Louis-Alexandre; Harel, Josée; Masson, Luke; Archambault, Marie

    2012-01-01

    Phenotypic bacitracin resistance has been reported in Clostridium perfringens. However, the genes responsible for the resistance have not yet been characterized. Ninety-nine C. perfringens isolates recovered from broilers and turkeys were tested for phenotypic bacitracin resistance. Bacitracin MIC90 (>256 µg/ml) was identical for both turkey and chicken isolates; whereas MIC50 was higher in turkey isolates (6 µg/ml) than in chicken isolates (3 µg/ml). Twenty-four of the 99 isolates showed high-level bacitracin resistance (MIC breakpoint >256 µg/ml) and the genes encoding for this resistance were characterized in C. perfringens c1261_A strain using primer walking. Sequence analysis and percentages of amino acid identity revealed putative genes encoding for both an ABC transporter and an overproduced undecaprenol kinase in C. perfringens c1261_A strain. These two mechanisms were shown to be both encoded by the putative bcrABD operon under the control of a regulatory gene, bcrR. Efflux pump inhibitor thioridazine was shown to increase significantly the susceptibility of strain c1261_A to bacitracin. Upstream and downstream from the bcr cluster was an IS1216-like element, which may play a role in the dissemination of this resistance determinant. Pulsed-field gel electrophoresis with prior double digestion with I-CeuI/MluI enzymes followed by hybridization analyses revealed that the bacitracin resistance genes bcrABDR were located on the chromosome. Semi-quantitative RT-PCR demonstrated that this gene cluster is expressed under bacitracin stress. Microarray analysis revealed the presence of these genes in all bacitracin resistant strains. This study reports the discovery of genes encoding for a putative ABC transporter and an overproduced undecaprenol kinase associated with high-level bacitracin resistance in C. perfringens isolates from turkeys and broiler chickens. PMID:22970221

  9. Transcriptional profiling analysis of host response to Clostridium perfringens infection in broilers.

    PubMed

    Zhou, H; Gong, J; Brisbin, J; Yu, H; Sarson, A J; Si, W; Sharif, S; Han, Y

    2009-05-01

    Necrotic enteritis is a disease caused by Clostridium perfringens, which threatens poultry production in the absence of dietary antibiotics. A total number of 600 Ross broilers were reared in 12 pens with each hosting 50 birds. Each 6 pens of birds were fed medicated (bacitracin at 55 mg/kg) or nonmedicated starter diets immediately after the chicks were placed. At d 18, birds were challenged with C. perfringens (10(7) cfu/mL mixed with feed). Spleens were collected from 12 birds of each group (2 birds per pen randomly) at d 18 (before infection), 19, 20, and 22. A low-density chicken immune microarray was used to study gene expression profiling of host response to C. perfringens infection. Six biological replicates (2 birds per biological replicate) for each treatment group were labeled with either Cy5 or Cy3 with dye swap. A total of 24 arrays were used for this study. Gene signal intensity was globally normalized by locally weighted regression and smoothing scatter plots and expressed on a natural log scale. A mixed model including treatment, time, array, subgrid (random effect), dye, and all interactions among treatment and time was used to identify differentially expressed genes between postinfection vs. preinfection, among postinfections, and between medication treatments, at the 5% significance level. The results indicated subtle medication effects on gene expression of these immune-related genes compared with bacterial infection effect. Our findings strongly suggest that both cell-mediated and antibody-mediated immune responses via MHC class I and II systems were actively involved in the host defense against C. perfringens infection in broilers. The unique cytokine signaling pathway and apoptosis cascade found in the study provide a new insight of molecular regulation of host immune response. Collectively, the findings of the present study will shed light on the molecular mechanisms underlying C. perfringens infection in broilers. PMID:19359691

  10. Clostridium perfringens: a flesh-eating bacterium living in your garden.

    PubMed

    Rothwell, Ann

    2010-10-01

    Gas gangrene is a painful, rapidly developing and potentially fatal infection despite antibiotic treatment. During the First World War thousands of soldiers died from this disease. Dr Alexis Carrel pioneered a controversial method of irrigating wounds with Dakin's solution to destroy Clostridium perfringens, a bacterium found in heavily fertilised soils that causes gas gangrene. Although this method is no longer used due to the discovery of antibiotics, many of his other ideas, such as scientifically determining the type and number of bacteria and delaying the closure of a wound until the bacteria had been eradicated, are still used today. PMID:21049805

  11. ADP-ribosylation of actin from the green alga Chara corallina by Clostridium botulinum C2 toxin and Clostridium perfringens iota toxin

    Microsoft Academic Search

    F. Grolig; I. Just; K. Aktories

    1996-01-01

    Summary The ability of two bacterial toxins to modify a plant actin by covalent ADP-ribosylation was tested in the green algaChara corallina. Using [32P]NAD, bothClostridium botulinum C2 toxin andClostridium perfringens iota toxin labelled a protein of Mr 42 kDa which comigrated with actin and was immunoprecipitated by a monoclonal anti-actin antibody. ADP-ribosylation ofChara actin was more efficient with iota toxin

  12. Characterization of Clostridium perfringens Spores That Lack SpoVA Proteins and Dipicolinic Acid?

    PubMed Central

    Paredes-Sabja, Daniel; Setlow, Barbara; Setlow, Peter; Sarker, Mahfuzur R.

    2008-01-01

    Spores of Clostridium perfringens possess high heat resistance, and when these spores germinate and return to active growth, they can cause gastrointestinal disease. Work with Bacillus subtilis has shown that the spore's dipicolinic acid (DPA) level can markedly influence both spore germination and resistance and that the proteins encoded by the spoVA operon are essential for DPA uptake by the developing spore during sporulation. We now find that proteins encoded by the spoVA operon are also essential for the uptake of Ca2+ and DPA into the developing spore during C. perfringens sporulation. Spores of a spoVA mutant had little, if any, Ca2+ and DPA, and their core water content was approximately twofold higher than that of wild-type spores. These DPA-less spores did not germinate spontaneously, as DPA-less B. subtilis spores do. Indeed, wild-type and spoVA C. perfringens spores germinated similarly with a mixture of l-asparagine and KCl (AK), KCl alone, or a 1:1 chelate of Ca2+ and DPA (Ca-DPA). However, the viability of C. perfringens spoVA spores was 20-fold lower than the viability of wild-type spores. Decoated wild-type and spoVA spores exhibited little, if any, germination with AK, KCl, or exogenous Ca-DPA, and their colony-forming efficiency was 103- to 104-fold lower than that of intact spores. However, lysozyme treatment rescued these decoated spores. Although the levels of DNA-protective ?/?-type, small, acid-soluble spore proteins in spoVA spores were similar to those in wild-type spores, spoVA spores exhibited markedly lower resistance to moist heat, formaldehyde, HCl, hydrogen peroxide, nitrous acid, and UV radiation than wild-type spores did. In sum, these results suggest the following. (i) SpoVA proteins are essential for Ca-DPA uptake by developing spores during C. perfringens sporulation. (ii) SpoVA proteins and Ca-DPA release are not required for C. perfringens spore germination. (iii) A low spore core water content is essential for full resistance of C. perfringens spores to moist heat, UV radiation, and chemicals. PMID:18469104

  13. Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test.

    PubMed Central

    Fach, P; Popoff, M R

    1997-01-01

    A duplex PCR procedure was evaluated for the detection of Clostridium perfringens in food and biological samples and for the identification of enterotoxigenic strains. This method uses two sets of primers which amplify in the same reaction two different DNA fragments simultaneously: the 283-bp C. perfringens phospholipase C gene fragment and the 426-bp enterotoxin gene fragment. Internal primers within the two primer sets confirmed the specificity of the method by DNA-DNA hybridization with the PCR products. No cross-reaction was observed with other Clostridium species or with other bacteria routinely found in food. The detection level was approximately 10(5) C. perfringens cells per g of stool or food sample. When overnight enrichment culture was used, 10 C. perfringens cells per g was detected in 57 artificially contaminated food samples. The duplex PCR is a rapid, sensitive, and reliable method for the detection and identification of enterotoxigenic C. perfringens strains in food samples. A slide latex agglutination test was also evaluated as a rapid, simple technique for the detection of C. perfringens enterotoxin in stool samples. PMID:9361409

  14. Antimicrobial susceptibility of Clostridium perfringens isolates of bovine, chicken, porcine, and turkey origin from Ontario.

    PubMed

    Slavi?, Dur?a; Boerlin, Patrick; Fabri, Marta; Klotins, Kim C; Zoethout, Jennifer K; Weir, Pat E; Bateman, Debbie

    2011-04-01

    Antimicrobial susceptibilities and toxin types were determined for 275 Clostridium perfringens isolates collected in Ontario in the spring of 2005. Minimal inhibitory concentrations (MICs) of C. perfringens isolates for 12 antimicrobials used in therapy, prophylaxis, and/or growth promotion of cattle (n = 40), swine (n = 75), turkeys (n = 50), and chickens (n = 100) were determined using the microbroth dilution method. Statistical analyses and MIC distributions showed reduced susceptibility to bacitracin, clindamycin, erythromycin, florfenicol, and tetracycline for some isolates. Reduced susceptibility to bacitracin was identified in chicken (64%) and turkey (60%) isolates. Swine isolates had predominantly reduced susceptibility to clindamycin (28%) and erythromycin (31%), whereas bovine isolates had reduced susceptibility to clindamycin (10%) and florfenicol (10%). Reduced susceptibility to tetracycline was spread across all species. No clear reduced susceptibility, but elevated MIC(50) for virginiamycin was found in chicken isolates in comparison with isolates from other species. Toxin typing revealed that C. perfringens type A is the dominant toxin type isolated in this study across all 4 host species. PMID:21731178

  15. Antibiotic resistance of Clostridium perfringens isolates from broiler chickens in Egypt.

    PubMed

    Osman, K M; Elhariri, M

    2013-12-01

    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 financially crippling poultry diseases, necrotic enteritis caused by Clostridium perfringens. The study was designed to determine the prevalence of antibiotic resistance in C. perfringens isolates from clinical cases of necrotic enteritis in broiler chickens in Egypt. A total of 125 isolates were obtained from broiler flocks in 35 chicken coops on 17 farms and were tested using the disc diffusion method. All 125 isolates were resistant to gentamicin, streptomycin, oxolinic acid, lincomycin, erythromycin and spiramycin. The prevalence of resistance to other antibiotics was also high: rifampicin (34%), chloramphenicol (46%), spectinomycin (50%), tylosin-fosfomycin (52%), ciprofloxacin (58%), norfloxacin (67%), oxytetracycline (71%), flumequine (78%), enrofloxacin (82%), neomycin (93%), colistin (94%), pefloxacin (94%), doxycycline (98%) and trimethoprim-sulfamethoxazole (98%). It is recommended that C. perfringens infections in Egypt should be treated with antibiotics for which resistant isolates are rare at present; namely, amoxicillin, ampicillin, cephradine, fosfomycin and florfenicol. PMID:24761735

  16. Antimicrobial susceptibility of Clostridium perfringens isolates of bovine, chicken, porcine, and turkey origin from Ontario

    PubMed Central

    Slavi?, ?ur?a; Boerlin, Patrick; Fabri, Marta; Klotins, Kim C.; Zoethout, Jennifer K.; Weir, Pat E.; Bateman, Debbie

    2011-01-01

    Antimicrobial susceptibilities and toxin types were determined for 275 Clostridium perfringens isolates collected in Ontario in the spring of 2005. Minimal inhibitory concentrations (MICs) of C. perfringens isolates for 12 antimicrobials used in therapy, prophylaxis, and/or growth promotion of cattle (n = 40), swine (n = 75), turkeys (n = 50), and chickens (n = 100) were determined using the microbroth dilution method. Statistical analyses and MIC distributions showed reduced susceptibility to bacitracin, clindamycin, erythromycin, florfenicol, and tetracycline for some isolates. Reduced susceptibility to bacitracin was identified in chicken (64%) and turkey (60%) isolates. Swine isolates had predominantly reduced susceptibility to clindamycin (28%) and erythromycin (31%), whereas bovine isolates had reduced susceptibility to clindamycin (10%) and florfenicol (10%). Reduced susceptibility to tetracycline was spread across all species. No clear reduced susceptibility, but elevated MIC50 for virginiamycin was found in chicken isolates in comparison with isolates from other species. Toxin typing revealed that C. perfringens type A is the dominant toxin type isolated in this study across all 4 host species. PMID:21731178

  17. Necrotic enteritis challenge models with broiler chickens raised on litter: evaluation of preconditions, Clostridium perfringens strains and outcome variables

    Microsoft Academic Search

    Magne Kaldhusdal; Merete Hofshagen; Atle Løvland; Haakon Langstrand; Keith Redhead

    1999-01-01

    The effect of Clostridium perfringens challenge, number of challenge days, and pre-challenge antibiotic treatment on the induction of necrotic enteritis in broiler chickens raised on litter was studied, and the relationship between bacterial counts and frequency of gut lesions was evaluated. Specific intestinal lesions in randomly selected birds were present despite a lack of disease-specific mortality. Challenge, number of challenge

  18. CONTROL OF CLOSTRIDIUM PERFRINGENS GERMINATION AND OUTGROWTH BY BUFFERED SODIUM CITRATE DURING CHILLING OF ROAST BEEF AND INJECTED PORK

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens germination and outgrowth by buffered sodium citrate and buffered sodium citrate supplemented with sodium diacetate was evaluated during abusive chilling of roast beef and injected pork. Beef top rounds and pork loins were injected with a brine containing NaCl (...

  19. CONTROL OF CLOSTRIDIUM PERFRINGENS GERMINATION AND OUTGROWTH BY BUFFERED SODIUM CITRATE DURING CHILLING OF ROAST BEEF AND INJECTED PORK.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens germination and outgrowth by buffered sodium citrate and buffered sodium citrate supplemented with sodium diacetate was evaluated during abusive chilling of roast beef and injected pork. Beef top rounds and pork loins were injected with a brine containing NaCl (...

  20. Control of Clostridium perfringens Spores by Green Tea Leaf Extracts During Cooling of Cooked Ground Beef, Chicken, and Pork

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We investigated the inhibition of Clostridium perfringens spore germination and outgrowth by two green tea extracts with low (GTL; 141 mg total catechins/g of green tea extract) and high (GTE; 697 mg total catechins/g of extract) catechin levels during abusive chilling of retail cooked ground beef, ...

  1. Effect of phosphate and meat (pork) types on the germination and outgrowth of Clostridium perfringens spores during abusive chilling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of blends of phosphates and the pork meat type (pale, soft and exudative, PSE; normal; and dark, firm and dry, DFD) on the germination and outgrowth of Clostridium perfringens during abusive exponential chilling times was evaluated. Two different phosphates, tetrasodium pyrophosphate (TSP...

  2. CARVACROL, CINNAMALDEHYDE, OREGANO OIL, AND THYMOL INHIBIT CLOSTRIDIUM PERFRINGENS SPORE GERMINATION AND OUTGROWTH IN GROUND TURKEY DURING CHILLING

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens by plant-derived carvacrol, cinnamaldehyde, thymol, and oregano oil was evaluated during abusive chilling of cooked ground turkey (75% lean) obtained from a local grocery store. Test substances were mixed into thawed turkey product at concentrations of 0.1, 0.5...

  3. Impedance Analysis of Ovarian Cancer Cells upon Challenge with C-terminal Clostridium Perfringens Enterotoxin

    NASA Astrophysics Data System (ADS)

    Gordon, Geoffrey; Lo, Chun-Min

    2007-03-01

    Both in vitro and animal studies in breast, prostate, and ovarian cancers have shown that clostridium perfringens enterotoxin (CPE), which binds to CLDN4, may have an important therapeutic benefit, as it is rapidly cytotoxic in tissues overexpressing CLDN4. This study sought to evaluate the ability of C-terminal clostridium perfringens enterotoxin (C-CPE), a CLDN4-targetting molecule, to disrupt tight junction barrier function. Electric cell-substrate impedance sensing (ECIS) was used to measure both junctional resistance and average cell-substrate separation of ovarian cancer cell lines after exposure to C-CPE. A total of 14 ovarian cancer cell lines were used, and included cell lines derived from serous, mucinous, and clear cells. Our results showed that junctional resistance increases as CLDN4 expression increases. In addition, C-CPE is non-cytotoxic in ovarian cancer cells expressing CLDN4. However, exposure to C-CPE results in a significant (p<0.05) dose- and CLDN4-dependent decrease in junctional resistance and an increase in cell-substrate separation. Treatment of ovarian cancer cell lines with C-CPE disrupts tight junction barrier function.

  4. Differential proteomic analysis of Clostridium perfringens ATCC13124; identification of dominant, surface and structure associated proteins

    PubMed Central

    2009-01-01

    Background Clostridium perfringens is a medically important clostridial pathogen causing diseases in man and animals. To invade, multiply and colonize tissues of the host, a pathogen must be able to evade host immune system, and obtain nutrients essential for growth. The factors involved in these complex processes are largely unknown and of crucial importance to understanding microbial pathogenesis. Many of the virulence determinants and putative vaccine candidates for bacterial pathogens are known to be surface localized. Results Using 2-DE mass spectrometry strategy, we identified major surface (22) and cell envelope (10) proteins from Clostridium perfringens ATCC13124 and those differentially expressed (11) in cells grown on cooked meat medium (CMM) in comparison with cells grown in reference state (tryptose-yeast extract-glucose medium). Riboflavin biosynthesis protein, ornithine carbamoyltransferase, cystathionine beta-lyase, and threonine dehydratase were the predominant proteins that exhibited 2.19 to 8.5 fold increase in the expression level in cells growing on CMM. Conclusion Ornithine carbamoyltransferase and cystathionine beta-lyase were over-expressed in cells grown on cooked meat medium and also identified in the surface protein fraction and the former was immunogenic; making them potential vaccine candidates. Based upon bioinformatic analysis; choloylglycine hydrolase family protein, cell wall-associated serine proteinase, and rhomboid family protein were predicted as surface protein markers for specific detection of C. perfringens from the environment and food. Most of the proteins over-expressed in CMM were shown to have putative function in metabolism, of which seven were involved in amino acid transport and metabolism or lipid metabolism. PMID:19664283

  5. Regulation of Sialidase Production in Clostridium perfringens by the Orphan Sensor Histidine Kinase ReeS

    PubMed Central

    Hiscox, Thomas J.; Harrison, Paul F.; Chakravorty, Anjana; Choo, Jocelyn M.; Ohtani, Kaori; Shimizu, Tohru

    2013-01-01

    Clostridium perfringens is ubiquitous in nature and is often found as a commensal of the human and animal gastrointestinal tract. It is the primary etiological agent of clostridial myonecrosis, or gas gangrene, a serious infection that results in extensive tissue necrosis due to the action of one or more potent extracellular toxins. ?-toxin and perfringolysin O are the major extracellular toxins involved in the pathogenesis of gas gangrene, but histotoxic strains of C. perfringens, such as strain 13, also produce many degradative enzymes such as collagenases, hyaluronidases, sialidases and the cysteine protease, ?-clostripain. The production of many of these toxins is regulated either directly or indirectly by the global VirSR two-component signal transduction system. By isolating a chromosomal mutant and carrying out microarray analysis we have identified an orphan sensor histidine kinase, which we have named ReeS (regulator of extracellular enzymes sensor). Expression of the sialidase genes nanI and nanJ was down-regulated in a reeS mutant. Since complementation with the wild-type reeS gene restored nanI and nanJ expression to wild-type levels, as shown by quantitative reverse transcription-PCR and sialidase assays we concluded that ReeS positively regulates the expression of these sialidase genes. However, mutation of the reeS gene had no significant effect on virulence in the mouse myonecrosis model. Sialidase production in C. perfringens has been previously shown to be regulated by both the VirSR system and RevR. In this report, we have analyzed a previously unknown sensor histidine kinase, ReeS, and have shown that it also is involved in controlling the expression of sialidase genes, adding further complexity to the regulatory network that controls sialidase production in C. perfringens. PMID:24023881

  6. Production of a bacteriocin by a poultry derived Campylobacter jejuni isolate with antimicrobial activity against Clostridium perfringens and other Gram positive bacteria.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have purified a bacteriocin peptide (termed CUV-3), produced by a poultry cecal isolate of Campylobacter jejuni (strain CUV-3) with inhibitory activity against Gram positive bacteria including Clostridium perfringens (38 strains), Staphylococcus aureus, Staphylococcus epidermidis and Listeria mon...

  7. Comparative Experiments To Examine the Effects of Heating on Vegetative Cells and Spores of Clostridium perfringens Isolates Carrying Plasmid Enterotoxin Genes versus Chromosomal Enterotoxin Genes

    Microsoft Academic Search

    MAHFUZUR R. SARKER; ROBERT P. SHIVERS; SHAUNA G. SPARKS; VIJAY K. JUNEJA; B. A. McClane

    2000-01-01

    Clostridium perfringens enterotoxin (CPE) is an important virulence factor for both C. perfringens type A food poisoning and several non-food-borne human gastrointestinal diseases. Recent studies have indicated that C. perfringens isolates associated with food poisoning carry a chromosomal cpe gene, while non-food-borne human gastrointestinal disease isolates carry a plasmid cpe gene. However, no explanation has been provided for the strong

  8. Evidence That the Enterotoxin Gene Can Be Episomal in Clostridium perfringens Isolates Associated with Non-Food-Borne Human Gastrointestinal Diseases

    Microsoft Academic Search

    RENEE E. COLLIE; BRUCE A. MCCLANE

    1998-01-01

    Clostridium perfringens enterotoxin (CPE) is responsible for the diarrheal and cramping symptoms of human C. perfringens type A food poisoning. CPE-producing C. perfringens isolates have also recently been associated with several non-food-borne human gastrointestinal (GI) illnesses, including antibiotic-associated diarrhea and sporadic diarrhea. The current study has used restriction fragment length polymorphism (RFLP) and pulsed-field gel electrophoresis (PFGE) analyses to compare

  9. Expression of a Clostridium perfringens genome-encoded putative N-acetylmuramoyl-L-alanine amidase as a potential antimicrobial to control the bacterium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium perfringens is a Gram-positive, spore-forming anaerobic bacterium that plays a substantial role in non-foodborne human, animal and avian diseases as well as human foodborne disease. Previously discovered C. perfringens bacteriophage lytic enzyme amino acid sequences were utilized to iden...

  10. Risk assessment for Clostridium perfringens in ready-to-eat and partially cooked meat and poultry products.

    PubMed

    Golden, Neal J; Crouch, Edmund A; Latimer, Heejeong; Kadry, Abdel-Razak; Kause, Janell

    2009-07-01

    An assessment of the risk of illness associated with Clostridium perfringens in ready-to-eat and partially cooked meat and poultry products was completed to estimate the effect on the annual frequency of illnesses of changing the allowed maximal 1-log growth of C. perfringens during stabilization (cooling after the manufacturing heat step). The exposure assessment modeled stabilization, storage, and consumer preparation such as reheating and hot-holding. The model predicted that assuming a 10- or 100-fold increase from the assumed 1-log (maximal allowable) growth of C. perfringens results in a 1.2- or 1.6-fold increase of C. perfringens-caused illnesses, respectively, at the median of the uncertainty distribution. Improper retail and consumer refrigeration accounted for approximately 90% of the 79,000 C. perfringens illnesses predicted by the model at 1-log growth during stabilization. Improper hot-holding accounted for 8% of predicted illnesses, although model limitations imply that this is an underestimate. Stabilization accounted for less than 1% of illnesses. Efforts to reduce illnesses from C. perfringens in ready-to-eat and partially cooked meat and poultry products should focus on retail and consumer storage and preparation methods. PMID:19681258

  11. The successful experimental induction of necrotic enteritis in chickens by Clostridium perfringens: a critical review

    PubMed Central

    2012-01-01

    Necrotic enteritis (NE) is one of the most important enteric diseases in poultry and is a high cost to the industry worldwide. It is caused by avian-specific, Necrotic Enteritis Beta toxin (NetB)-producing, strains of Clostridium perfringens that also possess in common other virulence-associated genes. In Europe the disease incidence has increased since the ban on in-feed “growth promoting” antibiotics. Because of this, many recent studies of NE have focused on finding different ways to control the disease, and on understanding its pathogenesis. Frustratingly, reproduction of the disease has proven impossible for some researchers. This review describes and discusses factors known to be important in reproducing the disease experimentally, as well as other considerations in reproducing the disease. The critical bacterial factor is the use of virulent, netB-positive, strains; virulence can be enhanced by using tpeL- positive strains and by the use of young rather than old broth cultures to increase toxin expression. Intestinal damaging factors, notably the use of concurrent or preceding coccidial infection, or administration of coccidial vaccines, combined with netB-positive C. perfringens administration, can also be used to induce NE. Nutritional factors, particularly feeding high percentage of cereals containing non-starch polysaccharides (NSP) (wheat, rye, and barley) enhance disease by increasing digesta viscosity, mucus production and bacterial growth. Animal proteins, especially fish meal, enhance C. perfringens proliferation and toxin production. Other factors are discussed that may affect outcome but for which evidence of their importance is lacking. The review compares the different challenge approaches; depending on the aim of particular studies, the different critical factors can be adjusted to affect the severity of the lesions induced. A standardized scoring system is proposed for international adoption based on gross rather than histopathological lesions; if universally adopted this will allow better comparison between studies done by different researchers. Also a scoring system is provided to assist decisions on humane euthanasia of sick birds. PMID:23101966

  12. Assessment of Clostridium perfringens spore response to high hydrostatic pressure and heat with nisin.

    PubMed

    Gao, Yulong; Qiu, Weifen; Wu, Ding; Fu, Qiang

    2011-08-01

    The elimination of spores from low-acid foods presents food-processing and food-safety challenges to high-pressure processing (HPP) developers as bacterial spores are extremely resistant to pressure. Therefore, the effects of pressure (400-800 MPa), temperature (35-95 °C), and nisin (0-496 IU/mL) on the inactivation of Clostridium perfringens AS 64701 spores at various pressure-holding times (7.5-17.5 min) were explored. A second-order polynomal equation for HPP- and nisin-induced inactivation of C. perfringens spores was constructed with response surface methodology. Experiment results showed that the experimental values were shown to be significantly in agreement with the predicted values because the adjusted determination coefficient (R (Adj)²) was 0.9708 and the level of significance was P?perfringens AS 64701 spores were pressure of 654 Mpa, temperature of 74 °C, pressure-holding time of 13.6 min, and nisin concentration of 328 IU/mL. The validation of the model equation for predicting the optimum response values was verified effectively by ten test points that were not used in the establishment of the model. Compared with conventional HPP techniques, the main process advantages of HPP-nisin combination sterilization in the UHT milk are, lower pressure, temperature, natural preservative (nisin), and in a shorter treatment time. The synergistic inactivation of bacteria by HPP-nisin combination is a promising and natural method to increase the efficiency and safety of high-pressure pasteurization. PMID:21340537

  13. Structural and biochemical analyses of a Clostridium perfringens sortase D transpeptidase

    PubMed Central

    Suryadinata, Randy; Seabrook, Shane A.; Adams, Timothy E.; Nuttall, Stewart D.; Peat, Thomas S.

    2015-01-01

    The assembly and anchorage of various pathogenic proteins on the surface of Gram-positive bacteria is mediated by the sortase family of enzymes. These cysteine transpeptidases catalyze a unique sorting signal motif located at the C-terminus of their target substrate and promote the covalent attachment of these proteins onto an amino nucleophile located on another protein or on the bacterial cell wall. Each of the six distinct classes of sortases displays a unique biological role, with sequential activation of multiple sortases often observed in many Gram-positive bacteria to decorate their peptidoglycans. Less is known about the members of the class D family of sortases (SrtD), but they have a suggested role in spore formation in an oxygen-limiting environment. Here, the crystal structure of the SrtD enzyme from Clostridium perfringens was determined at 1.99?Å resolution. Comparative analysis of the C. perfringens SrtD structure reveals the typical eight-stranded ?-barrel fold observed in all other known sortases, along with the conserved catalytic triad consisting of cysteine, histidine and arginine residues. Biochemical approaches further reveal the specifics of the SrtD catalytic activity in vitro, with a significant preference for the LPQTGS sorting motif. Additionally, the catalytic activity of SrtD is most efficient at 316?K and can be further improved in the presence of magnesium cations. Since C. perfringens spores are heat-resistant and lead to foodborne illnesses, characterization of the spore-promoting sortase SrtD may lead to the development of new antimicrobial agents. PMID:26143922

  14. Identification of Novel Pathogenicity Loci in Clostridium perfringens Strains That Cause Avian Necrotic Enteritis

    PubMed Central

    Parreira, Valeria R.; Marri, Pradeep R.; Rosey, Everett L.; Gong, Joshua; Songer, J. Glenn; Vedantam, Gayatri; Prescott, John F.

    2010-01-01

    Type A Clostridium perfringens causes poultry necrotic enteritis (NE), an enteric disease of considerable economic importance, yet can also exist as a member of the normal intestinal microbiota. A recently discovered pore-forming toxin, NetB, is associated with pathogenesis in most, but not all, NE isolates. This finding suggested that NE-causing strains may possess other virulence gene(s) not present in commensal type A isolates. We used high-throughput sequencing (HTS) technologies to generate draft genome sequences of seven unrelated C. perfringens poultry NE isolates and one isolate from a healthy bird, and identified additional novel NE-associated genes by comparison with nine publicly available reference genomes. Thirty-one open reading frames (ORFs) were unique to all NE strains and formed the basis for three highly conserved NE-associated loci that we designated NELoc-1 (42 kb), NELoc-2 (11.2 kb) and NELoc-3 (5.6 kb). The largest locus, NELoc-1, consisted of netB and 36 additional genes, including those predicted to encode two leukocidins, an internalin-like protein and a ricin-domain protein. Pulsed-field gel electrophoresis (PFGE) and Southern blotting revealed that the NE strains each carried 2 to 5 large plasmids, and that NELoc-1 and -3 were localized on distinct plasmids of sizes ?85 and ?70 kb, respectively. Sequencing of the regions flanking these loci revealed similarity to previously characterized conjugative plasmids of C. perfringens. These results provide significant insight into the pathogenetic basis of poultry NE and are the first to demonstrate that netB resides in a large, plasmid-encoded locus. Our findings strongly suggest that poultry NE is caused by several novel virulence factors, whose genes are clustered on discrete pathogenicity loci, some of which are plasmid-borne. PMID:20532244

  15. Identification of novel pathogenicity loci in Clostridium perfringens strains that cause avian necrotic enteritis.

    PubMed

    Lepp, Dion; Roxas, Bryan; Parreira, Valeria R; Marri, Pradeep R; Rosey, Everett L; Gong, Joshua; Songer, J Glenn; Vedantam, Gayatri; Prescott, John F

    2010-01-01

    Type A Clostridium perfringens causes poultry necrotic enteritis (NE), an enteric disease of considerable economic importance, yet can also exist as a member of the normal intestinal microbiota. A recently discovered pore-forming toxin, NetB, is associated with pathogenesis in most, but not all, NE isolates. This finding suggested that NE-causing strains may possess other virulence gene(s) not present in commensal type A isolates. We used high-throughput sequencing (HTS) technologies to generate draft genome sequences of seven unrelated C. perfringens poultry NE isolates and one isolate from a healthy bird, and identified additional novel NE-associated genes by comparison with nine publicly available reference genomes. Thirty-one open reading frames (ORFs) were unique to all NE strains and formed the basis for three highly conserved NE-associated loci that we designated NELoc-1 (42 kb), NELoc-2 (11.2 kb) and NELoc-3 (5.6 kb). The largest locus, NELoc-1, consisted of netB and 36 additional genes, including those predicted to encode two leukocidins, an internalin-like protein and a ricin-domain protein. Pulsed-field gel electrophoresis (PFGE) and Southern blotting revealed that the NE strains each carried 2 to 5 large plasmids, and that NELoc-1 and -3 were localized on distinct plasmids of sizes approximately 85 and approximately 70 kb, respectively. Sequencing of the regions flanking these loci revealed similarity to previously characterized conjugative plasmids of C. perfringens. These results provide significant insight into the pathogenetic basis of poultry NE and are the first to demonstrate that netB resides in a large, plasmid-encoded locus. Our findings strongly suggest that poultry NE is caused by several novel virulence factors, whose genes are clustered on discrete pathogenicity loci, some of which are plasmid-borne. PMID:20532244

  16. Structural and biochemical analyses of a Clostridium perfringens sortase D transpeptidase.

    PubMed

    Suryadinata, Randy; Seabrook, Shane A; Adams, Timothy E; Nuttall, Stewart D; Peat, Thomas S

    2015-07-01

    The assembly and anchorage of various pathogenic proteins on the surface of Gram-positive bacteria is mediated by the sortase family of enzymes. These cysteine transpeptidases catalyze a unique sorting signal motif located at the C-terminus of their target substrate and promote the covalent attachment of these proteins onto an amino nucleophile located on another protein or on the bacterial cell wall. Each of the six distinct classes of sortases displays a unique biological role, with sequential activation of multiple sortases often observed in many Gram-positive bacteria to decorate their peptidoglycans. Less is known about the members of the class D family of sortases (SrtD), but they have a suggested role in spore formation in an oxygen-limiting environment. Here, the crystal structure of the SrtD enzyme from Clostridium perfringens was determined at 1.99?Å resolution. Comparative analysis of the C. perfringens SrtD structure reveals the typical eight-stranded ?-barrel fold observed in all other known sortases, along with the conserved catalytic triad consisting of cysteine, histidine and arginine residues. Biochemical approaches further reveal the specifics of the SrtD catalytic activity in vitro, with a significant preference for the LPQTGS sorting motif. Additionally, the catalytic activity of SrtD is most efficient at 316?K and can be further improved in the presence of magnesium cations. Since C. perfringens spores are heat-resistant and lead to foodborne illnesses, characterization of the spore-promoting sortase SrtD may lead to the development of new antimicrobial agents. PMID:26143922

  17. Identification of bilirubin reduction products formed by Clostridium perfringens isolated from human neonatal fecal flora.

    PubMed

    Vítek, Libor; Majer, Filip; Muchová, Lucie; Zelenka, Jaroslav; Jirásková, Alena; Branný, Pavel; Malina, Jirí; Ubik, Karel

    2006-04-01

    Urobilinoids belong to the heterogenous group of degradation products of bilirubin formed in the gastrointestinal tract by intestinal microflora. Among them urobilinogen and stercobilinogen with their respective oxidation products, urobilin and stercobilin, are the most important compounds. The aim of present study was to analyze the products of bacterial reduction of bilirubin in more detail. The strain of Clostridium perfringens isolated from neonatal stools, capable of reducing bilirubin, was used in the study. Bacteria were incubated under anaerobic conditions with various native as well as synthetic bile pigments, including radiolabeled unconjugated bilirubin (UCB). Their reduction products were extracted from media and separated following thin layer chromatography. Pigments isolated were analyzed by spectrophotometry, spectrofluorometry and mass spectrometry. In a special set of experiments, bilirubin diglucuronide was incubated with either bacterial lysate or partially purified bilirubin reductase and beta-glucuronidase to reveal whether bilirubin glucuronides may be directly reduced onto conjugated urobilinoids. A broad substrate activity was detected in the investigated strain of C. perfringens and a series of bilirubin reduction products was identified. These products were separated in the form of their respective chromogens and further oxidized. Based on their physical-chemical properties, as well as mass spectra, end-catabolic bilirubin products were identified to belong to urobilinogen species. The reduction process, catalyzed enzymatically by the studied bacterial strain, does not proceed to stercobilinogen. Bilirubin diglucuronide is not reduced onto urobilinoid conjugates, glucuronide hydrolysis must precede double bond reduction and thus UCB is reduced much faster. PMID:16504607

  18. The Structure of Clostridium perfringens NanI Sialidase and Its Catalytic Intermediates*

    PubMed Central

    Newstead, Simon L.; Potter, Jane A.; Wilson, Jennifer C.; Xu, Guogang; Chien, Chin-Hsiang; Watts, Andrew G.; Withers, Stephen G.; Taylor, Garry L.

    2008-01-01

    Clostridium perfringens is a Gram-positive bacterium responsible for bacteremia, gas gangrene, and occasionally food poisoning. Its genome encodes three sialidases, nanH, nanI, and nanJ, that are involved in the removal of sialic acids from a variety of glycoconjugates and that play a role in bacterial nutrition and pathogenesis. Recent studies on trypanosomal (trans-) sialidases have suggested that catalysis in all sialidases may proceed via a covalent intermediate similar to that of other retaining glycosidases. Here we provide further evidence to support this suggestion by reporting the 0.97Å resolution atomic structure of the catalytic domain of the C. perfringens NanI sialidase, and complexes with its substrate sialic acid (N-acetylneuramic acid) also to 0.97Å resolution, with a transition-state analogue (2-deoxy-2,3-dehydro-N-acetylneuraminic acid) to 1.5Å resolution, and with a covalent intermediate formed using a fluorinated sialic acid analogue to 1.2Å resolution. Together, these structures provide high resolution snapshots along the catalytic pathway. The crystal structures suggested that NanI is able to hydrate 2-deoxy-2,3-dehydro-N-acetylneuraminic acid to N-acetylneuramic acid. This was confirmed by NMR, and a mechanism for this activity is suggested. PMID:18218621

  19. The structure of Clostridium perfringens NanI sialidase and its catalytic intermediates.

    PubMed

    Newstead, Simon L; Potter, Jane A; Wilson, Jennifer C; Xu, Guogang; Chien, Chin-Hsiang; Watts, Andrew G; Withers, Stephen G; Taylor, Garry L

    2008-04-01

    Clostridium perfringens is a Gram-positive bacterium responsible for bacteremia, gas gangrene, and occasionally food poisoning. Its genome encodes three sialidases, nanH, nanI, and nanJ, that are involved in the removal of sialic acids from a variety of glycoconjugates and that play a role in bacterial nutrition and pathogenesis. Recent studies on trypanosomal (trans-) sialidases have suggested that catalysis in all sialidases may proceed via a covalent intermediate similar to that of other retaining glycosidases. Here we provide further evidence to support this suggestion by reporting the 0.97A resolution atomic structure of the catalytic domain of the C. perfringens NanI sialidase, and complexes with its substrate sialic acid (N-acetylneuramic acid) also to 0.97A resolution, with a transition-state analogue (2-deoxy-2,3-dehydro-N-acetylneuraminic acid) to 1.5A resolution, and with a covalent intermediate formed using a fluorinated sialic acid analogue to 1.2A resolution. Together, these structures provide high resolution snapshots along the catalytic pathway. The crystal structures suggested that NanI is able to hydrate 2-deoxy-2,3-dehydro-N-acetylneuraminic acid to N-acetylneuramic acid. This was confirmed by NMR, and a mechanism for this activity is suggested. PMID:18218621

  20. Purification, characterization, and primary structure of Clostridium perfringens lambda-toxin, a thermolysin-like metalloprotease.

    PubMed Central

    Jin, F; Matsushita, O; Katayama, S; Jin, S; Matsushita, C; Minami, J; Okabe, A

    1996-01-01

    The lambda-toxin of Clostridium perfringens type B NCIB10691 was purified by ammonium sulfate precipitation, followed by size exclusion, anion-exchange, and hydrophobic interaction chromatography. The purified toxin had an apparent molecular mass of 36 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The toxin possessed casein-hydrolyzing activity, which was inhibited specifically by metal chelators, indicating that the toxin is a metalloprotease. The gene encoding the lambda-toxin (lam), which was shown by Southern analysis to be located on a 70-kb plasmid, was cloned into Escherichia coli cells. Nucleotide and N-terminal amino acid sequencing revealed that the lam gene encodes a 553-amino-acid protein, which is processed into a mature form, the molecular mass of which was calculated to be 35,722 Da. The deduced amino acid sequence of the mature enzyme contains an HEXXH motif characteristic of zinc metalloproteases and is homologous to other known enzymes belonging to the thermolysin family. The purified toxin degraded various biologically important substances, such as collagen, fibronectin, fibrinogen, immunoglobulin A, and the complement C3 component. It caused an increase in vascular permeability and hemorrhagic edema on injection into the dorsal skin of mice. These results suggest that the toxin contributes to the pathogenesis of histolytic infection by lambda-toxin-producing C. perfringens. PMID:8557345

  1. A Paracrystalline Inclusion Formed During Sporulation of Enterotoxin-Producing Strains of Clostridium perfringens Type A

    PubMed Central

    Duncan, Charles L.; King, Gretchen J.; Frieben, William R.

    1973-01-01

    A large paracrystalline inclusion is formed by certain strains of Clostridium perfringens type A during spore morphogenesis. In most cell thin sections, the inclusion appeared rod-shaped when sectioned at an angle perpendicular to its longer axis, and circular or oval-shaped when sectioned at an angle parallel to its longer axis. Measurements performed on electron micrographs of inclusions sectioned to reveal the rod shape indicated a fairly consistent thickness (width) of 192 ± 23 nm. The length of the inclusions varied considerably with a maximum of approximately 2,120 nm being observed. Ultrastructurally, the inclusion was composed of closely packed, periodically spaced, parallel layers. Usually a single inclusion was randomly located in the cytoplasm of the cell. Two inclusions per cell were rarely observed. The inclusion was formed only by ent+ strains of C. perfringens. Mutants of the ent+ strain NCTC 8798 that were altered in their sporulating and enterotoxin-producing capacities and revertants of these mutants were tested for inclusion formation. The results indicate that, as with the ent+ trait, a direct relationship exists between inclusion formation and spore formation. The synthesis of enterotoxin, formation of a morphologically distinct inclusion, and the initial deposition of discontinuous coat fragments around the forespore appear to be events closely related in time during spore morphogenesis. Images PMID:4350345

  2. Mechanism of Aminoglycoside Antibiotic Resistance in Anaerobic Bacteria: Clostridium perfringens and Bacteroides fragilis

    PubMed Central

    Bryan, L. E.; Kowand, S. K.; Van Den Elzen, H. M.

    1979-01-01

    Cell-free amino acid incorporation using ribosomes from strains of either Clostridium perfringens or Bacteroides fragilis was shown to be susceptible to inhibition by streptomycin and gentamicin. Ribosomes bound dihydrostreptomycin as effectively as ribosomes from Escherichia coli. No inactivation of streptomycin or gentamicin was detected by cell extracts of either anaerobic bacterial species. B. fragilis, grown without added hemin, menadione, and fumarate, and C. perfringens did not show any time-dependent accumulation of dihydrostreptomycin or gentamicin at concentrations tested. Decreased resistance to aminoglycosides and time-dependent uptake of dihydrostreptomycin at 500 ?g/ml was observed with B. fragilis grown with hemin, menadione, and fumarate. With the last additions, cytochrome b was detected by cytochrome spectra of B. fragilis. These results demonstrate that anaerobic bacteria unable to carry out oxygen- or nitrate-dependent electron transport are resistant to streptomycin and gentamicin because of failure to transport aminoglycosides. The induction of fumarate-dependent electron transport in B. fragilis is associated with some aminoglycoside transport that is of poor efficiency relative to bacteria with electron transport to oxygen or nitrate. PMID:218500

  3. Identification of a two-component VirR/VirS regulon in Clostridium perfringens.

    PubMed

    Ohtani, Kaori; Hirakawa, Hideki; Tashiro, Kousuke; Yoshizawa, Satoko; Kuhara, Satoru; Shimizu, Tohru

    2010-06-01

    Clostridium perfringens, a Gram-positive anaerobic pathogen, is a causative agent of human gas gangrene that leads to severe rapid tissue destruction and can cause death within hours unless treated immediately. Production of several toxins is known to be controlled by the two-component VirR/VirS system involving a regulatory RNA (VR-RNA) in C. perfringens. To elucidate the precise regulatory network governed by VirR/VirS and VR-RNA, a series of microarray screening using VirR/VirS and VR-RNA-deficient mutants was performed. Finally, by qRT-PCR analysis, 147 genes (30 single genes and 21 putative operons) were confirmed to be under the control of the VirR/VirS-VR-RNA regulatory cascade. Several virulence-related genes for alpha-toxin, kappa-toxin, hyaluronidases, sialidase, and capsular polysaccharide synthesis were found. Furthermore, some genes for catalytic enzymes, various genes for transporters, and many genes for energy metabolism were also found to be controlled by the cascade. Our data indicate that the VirR/VirS-VR-RNA system is a global gene regulator that might control multiple cellular functions to survive and multiply in the host, which would turn out to be a lethal flesh-eating infection. PMID:19835966

  4. Clostridium perfringens bacteriophages ?CP39O and ?CP26F: genomic organization and proteomic analysis of the virions

    Microsoft Academic Search

    Bruce S. Seal; Derrick E. Fouts; Mustafa Simmons; Johnna K. Garrish; Robin L. Kuntz; Rebekah Woolsey; Kathleen M. Schegg; Andrew M. Kropinski; Hans-W. Ackermann; Gregory R. Siragusa

    2011-01-01

    Poultry intestinal material, sewage and poultry processing drainage water were screened for virulent Clostridium perfringens bacteriophages. Viruses isolated from broiler chicken offal washes (O) and poultry feces (F), designated ?CP39O and ?CP26F,\\u000a respectively, produced clear plaques on host strains. Both bacteriophages had isometric heads of 57 nm in diameter with 100-nm\\u000a non-contractile tails characteristic of members of the family Siphoviridae in

  5. Identification and Characterization of Sporulation-Dependent Promoters Upstream of the Enterotoxin Gene (cpe )o f Clostridium perfringens

    Microsoft Academic Search

    YULING ZHAO; STEPHEN B. MELVILLE

    1998-01-01

    Three promoter sites (P1, P2, and P3) responsible for the sporulation-associated synthesis of Clostridium perfringens enterotoxin, a common cause of food poisoning in humans and animals, were identified. Nested and internal deletions of the cpe promoter region were made to narrow down the location of promoter elements. To measure the effects of the deletions on the expression of cpe, translational

  6. The Mycotoxin Deoxynivalenol Predisposes for the Development of Clostridium perfringens-Induced Necrotic Enteritis in Broiler Chickens

    PubMed Central

    Antonissen, Gunther; Ducatelle, Richard; Haesebrouck, Freddy; Timbermont, Leen; Verlinden, Marc; Janssens, Geert Paul Jules; Eeckhaut, Venessa; Eeckhout, Mia; De Saeger, Sarah; Hessenberger, Sabine; Martel, An; Croubels, Siska

    2014-01-01

    Both mycotoxin contamination of feed and Clostridium perfringens-induced necrotic enteritis have an increasing global economic impact on poultry production. Especially the Fusarium mycotoxin deoxynivalenol (DON) is a common feed contaminant. This study aimed at examining the predisposing effect of DON on the development of necrotic enteritis in broiler chickens. An experimental Clostridium perfringens infection study revealed that DON, at a contamination level of 3,000 to 4,000 µg/kg feed, increased the percentage of birds with subclinical necrotic enteritis from 20±2.6% to 47±3.0% (P<0.001). DON significantly reduced the transepithelial electrical resistance in duodenal segments (P<0.001) and decreased duodenal villus height (P?=?0.014) indicating intestinal barrier disruption and intestinal epithelial damage, respectively. This may lead to an increased permeability of the intestinal epithelium and decreased absorption of dietary proteins. Protein analysis of duodenal content indeed showed that DON contamination resulted in a significant increase in total protein concentration (P?=?0.023). Furthermore, DON had no effect on in vitro growth, alpha toxin production and netB toxin transcription of Clostridium perfringens. In conclusion, feed contamination with DON at concentrations below the European maximum guidance level of 5,000 µg/kg feed, is a predisposing factor for the development of necrotic enteritis in broilers. These results are associated with a negative effect of DON on the intestinal barrier function and increased intestinal protein availability, which may stimulate growth and toxin production of Clostridium perfringens. PMID:25268498

  7. Characterization of an outbreak of Clostridium perfringens food poisoning by quantitative fecal culture and fecal enterotoxin measurement.

    PubMed Central

    Birkhead, G; Vogt, R L; Heun, E M; Snyder, J T; McClane, B A

    1988-01-01

    Published criteria for implicating Clostridium perfringens as the cause of food-poisoning outbreaks include finding a median fecal C. perfringens spore count of greater than 10(6)/g among specimens from ill persons. We investigated a food-poisoning outbreak with the epidemiologic characteristics of C. perfringens-related disease in a nursing home in which the median fecal spore count for ill patients (2.5 X 10(7)/g) was similar to that for well patients (4.0 X 10(6)/g), making the etiology of the outbreak uncertain. All ill and well patients tested had eaten turkey, the implicated food item. C. perfringens enterotoxin was detected by reverse passive latex agglutination in fecal specimens from six of six ill and none of four well patients who had eaten turkey (P = 0.005), suggesting that this organism had caused the outbreak. This investigation suggests that detection of fecal C. perfringens enterotoxin is a specific way to identify this organism as the causative agent in food-poisoning outbreaks. PMID:2895776

  8. Rapid, Simultaneous Detection of Clostridium sordellii and Clostridium perfringens in Archived Tissues by a Novel PCR-Based Microsphere Assay: Diagnostic Implications for Pregnancy-Associated Toxic Shock Syndrome Cases

    PubMed Central

    Bhatnagar, Julu; DeLeon-Carnes, Marlene; Kellar, Kathryn L.; Bandyopadhyay, Kakali; Antoniadou, Zoi-Anna; Shieh, Wun-Ju; Paddock, Christopher D.; Zaki, Sherif R.

    2012-01-01

    Clostridium sordellii and Clostridium perfringens are infrequent human pathogens; however, the case-fatality rates for the infections are very high, particularly in obstetric C. sordellii infections (>90%). Deaths from Clostridium sordellii and Clostridium perfringens toxic shock (CTS) are sudden, and diagnosis is often challenging. Formalin-fixed, paraffin-embedded (FFPE) tissues usually are the only specimens available for sudden fatal cases, and immunohistochemistry (IHC) for Clostridia is generally performed but it cannot identify species. A clear need exists for a rapid, species-specific diagnostic assay for FFPE tissues. We developed a duplex PCR-based microsphere assay for simultaneous detection of C. sordellii and C. perfringens and evaluated DNA extracted from 42 Clostridium isolates and FFPE tissues of 28 patients with toxic shock/endometritis (20?CTS, 8?non-CTS, as confirmed by PCR and sequencing). The microsphere assay correctly identified C. sordellii and C. perfringens in all known isolates and in all CTS patients (10 C. sordellii, 8 C. perfringens, 2 both) and showed 100% concordance with PCR and sequencing results. The microsphere assay is a rapid, specific, and cost-effective method for the diagnosis of CTS and offers the advantage of simultaneous testing for C. sordellii and C. perfringens in FFPE tissues using a limited amount of DNA. PMID:22536012

  9. A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability.

    PubMed

    Swift, Steven M; Seal, Bruce S; Garrish, Johnna K; Oakley, Brian B; Hiett, Kelli; Yeh, Hung-Yueh; Woolsey, Rebekah; Schegg, Kathleen M; Line, John Eric; Donovan, David M

    2015-01-01

    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 gene was synthesized that fused the N-acetylmuramoyl-L-alanine amidase domain from the endolysin of the thermophilic bacteriophage ?GVE2 to the cell-wall binding domain (CWB) from the endolysin of the C. perfringens-specific bacteriophage ?CP26F. The resulting protein, PlyGVE2CpCWB, lysed C. perfringens in liquid and solid cultures. PlyGVE2CpCWB was most active at pH 8, had peak activity at 10 mM NaCl, 40% activity at 150 mM NaCl and was still 16% active at 600 mM NaCl. The protein was able to withstand temperatures up to 50° C and still lyse C. perfringens. Herein, we report the construction and characterization of a thermostable chimeric endolysin that could potentially be utilized as a feed additive to control the bacterium during poultry production. PMID:26075507

  10. A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability

    PubMed Central

    Swift, Steven M.; Seal, Bruce S.; Garrish, Johnna K.; Oakley, Brian B.; Hiett, Kelli; Yeh, Hung-Yueh; Woolsey, Rebekah; Schegg, Kathleen M.; Line, John Eric; Donovan, David M.

    2015-01-01

    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 gene was synthesized that fused the N-acetylmuramoyl-l-alanine amidase domain from the endolysin of the thermophilic bacteriophage ?GVE2 to the cell-wall binding domain (CWB) from the endolysin of the C. perfringens-specific bacteriophage ?CP26F. The resulting protein, PlyGVE2CpCWB, lysed C. perfringens in liquid and solid cultures. PlyGVE2CpCWB was most active at pH 8, had peak activity at 10 mM NaCl, 40% activity at 150 mM NaCl and was still 16% active at 600 mM NaCl. The protein was able to withstand temperatures up to 50 °C and still lyse C. perfringens. Herein, we report the construction and characterization of a thermostable chimeric endolysin that could potentially be utilized as a feed additive to control the bacterium during poultry production. PMID:26075507

  11. Clostridium perfringens in poultry: an emerging threat for animal and public health

    Microsoft Academic Search

    Filip Van Immerseel; Jeroen De Buck; Frank Pasmans; Gerard Huyghebaert; Freddy Haesebrouck; Richard Ducatelle

    2004-01-01

    The incidence of Clostridiumperfringens-associated necrotic enteritis in poultry has increased in countries that stopped using antibiotic growth promoters. Necrotic enteritis and the subclinical form of C. perfringens infection in poultry are caused by C. perfringens type A, producing the alpha toxin, and to a lesser extent type C, producing both alpha toxin and beta toxin. Some strains of C. perfringens

  12. Clostridium perfringens Is Not Suitable for the Indication of Fecal Pollution from Ruminant Wildlife but Is Associated with Excreta from Nonherbivorous Animals and Human Sewage

    PubMed Central

    Vierheilig, J.; Frick, C.; Mayer, R. E.; Kirschner, A. K. T.; Reischer, G. H.; Derx, J.; Mach, R. L.; Farnleitner, A. H.

    2013-01-01

    During a 3-year study, Clostridium perfringens was investigated in defined fecal sources from a temperate alluvial backwater area of a large river system. The results reveal that using C. perfringens as a conservative water quality indicator for total fecal pollution monitoring is no longer justified but suggest that it can be used as a tracer for excreta from nonherbivorous wildlife and human sewage. PMID:23747707

  13. Multiple effects of Escherichia coli Nissle 1917 on growth, biofilm formation and inflammation cytokines profile of Clostridium perfringens type A strain CP4

    PubMed Central

    Jiang, Yanlong; Kong, Qingke; Roland, Kenneth L.; Wolf, Amanda; Curtiss, Roy

    2014-01-01

    Clostridium perfringens is an important Gram-positive pathogen responsible for food poisoning, necrotic enteritis, gas gangrene, and even death. Escherichia coli Nissle 1917 (EcN) is a well-characterized probiotic strain with demonstrated benefits. In this study we evaluated the effects of EcN on growth, toxin production, biofilm formation and inflammatory cytokine responses of C. perfringens. In vitro co-culture experiments demonstrated that EcN inhibited growth, gas production and toxin production (?-toxin and NetB) of C. perfringens in a dose dependent manner. The growth inhibition effect was not observed when C. perfringens was incubated with EcN cell free supernatants (CFSE), suggesting that growth inhibition was caused by nutrition competition during co-incubation. In vitro studies demonstrated that pre-incubation with EcN did not inhibit C. perfringens attachment to Caco-2 cells, but did reduce C. perfringens total number, toxin production and cytotoxicity after 24 h. The similar growth inhibition results were also observed during the formation of C. perfringens biofilm. Finally, pre-incubation of EcN with RAW264.7 cells significantly decreased the production of inflammatory cytokines caused by introduction of C. perfringens. Our results indicate that EcN can inhibit many of the pathological effects of C. perfringens in vitro conditions. PMID:24532573

  14. C-Terminal Clostridium perfringens Enterotoxin-Mediated Antigen Delivery for Nasal Pneumococcal Vaccine

    PubMed Central

    Suzuki, Hidehiko; Watari, Akihiro; Hashimoto, Eri; Yonemitsu, Miki; Kiyono, Hiroshi; Yagi, Kiyohito; Kondoh, Masuo; Kunisawa, Jun

    2015-01-01

    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 applied the C-CPE-based vaccine delivery system to develop a nasal pneumococcal vaccine. We fused C-CPE with pneumococcal surface protein A (PspA), an important antigen for the induction of protective immunity against Streptococcus pneumoniae infection, (PspA-C-CPE). PspA-C-CPE binds to claudin-4 and thus efficiently attaches to NALT epithelium, including antigen-sampling M cells. Nasal immunization with PspA-C-CPE induced PspA-specific IgG in the serum and bronchoalveolar lavage fluid (BALF) as well as IgA in the nasal wash and BALF. These immune responses were sufficient to protect against pneumococcal infection. These results suggest that C-CPE is an efficient vaccine delivery system for the development of nasal vaccines against pneumococcal infection. PMID:26018248

  15. Intestinal events and nutritional dynamics predispose Clostridium perfringens virulence in broilers.

    PubMed

    Moran, Edwin T

    2014-12-01

    Clostridium perfringensA (CPA) entering the gastrointestinal system depends on favorable conditions to develop and subsequently extend pathogenicity. Reduction in digestive dynamics progressing from the duodenum decreases lumen oxygen, leading to anaerobic conditions in the distal lumen that favor CPA. When nutritional support is concurrently provided, an expanding population threatens the mucosa. Dietary nonstarch polysaccharides that increase viscosity further impair oxygen transfer from the mucosa, improving the ability of CPA to thrive. Incompletion of feed digestion early in the small intestine along with endogenous N provide additional support for population expansion. Glucosidase versatility with mucin elicited by distal CPA concurrently erodes the villus unstirred water layer at the apex, providing access to underlying binding sites for colonization. Proteolytic destruction within the lamina propria supports colonization to create subclinical necrotic enteritis. Eventual vascular entry of CPA and toxins provides a portal path for instituting cholangiohepatitis. Liver condemnations from inspection detect acute flock infection compared with preceding marginal losses in nutrient absorption that decrease feed efficiency. Enterocyte lysis by coccidia enable CPA access to binding sites, thereby extending villus necrosis and further impairing feed conversion. Loss of BW and increased mortality follow as mucosa involvement proceeds. In practice, supplemental feed hemicellulases that reduce digesta viscosity minimize a favorable environment for CPA, while superimposing a combination of amylase, phytase, and protease avoids nutritional support. Physical dynamics of the small intestine together with characteristics of feed that modify digesta viscosity and nutritional availability are central to establishing transient CPA as a pathogen. PMID:25260526

  16. Inactivation and ultrastructure analysis of Bacillus spp. and Clostridium perfringens spores.

    PubMed

    Brantner, Christine A; Hannah, Ryan M; Burans, James P; Pope, Robert K

    2014-02-01

    Bacterial endospores are resistant to many environmental factors from temperature extremes to ultraviolet irradiation and are generally more difficult to inactivate or kill than vegetative bacterial cells. It is often considered necessary to treat spores or samples containing spores with chemical fixative solutions for prolonged periods of time (e.g., 1-21 days) to achieve fixation/inactivation to enable electron microscopy (EM) examination outside of containment laboratories. Prolonged exposure to chemical fixatives, however, can alter the ultrastructure of spores for EM analyses. This study was undertaken to determine the minimum amount of time required to inactivate/sterilize and fix spore preparations from several bacterial species using a universal fixative solution for EM that maintains the ultrastructural integrity of the spores. We show that a solution of 4% paraformaldehyde with 1% glutaraldehyde inactivated spore preparations of Bacillus anthracis, Bacillus cereus, Bacillus megaterium, Bacillus thuringiensis, and Clostridium perfringens in 30 min, and Bacillus subtilis in 240 min. These results suggest that this fixative solution can be used to inactivate and fix spores from several major groups of bacterial spore formers after 240 min, enabling the fixed preparations to be removed from biocontainment and safely analyzed by EM outside of biocontainment. PMID:24503289

  17. Fast kinetics of nucleotide binding to Clostridium perfringens family II pyrophosphatase containing CBS and DRTGG domains.

    PubMed

    Jämsen, J; Baykov, A A; Lahti, R

    2012-02-01

    We earlier described CBS-pyrophosphatase of Moorella thermoacetica (mtCBS-PPase) as a novel phosphohydrolase that acquired a pair of nucleotide-binding CBS domains during evolution, thus endowing the protein with the capacity to be allosterically regulated by adenine nucleotides (Jämsen, J., Tuominen, H., Salminen, A., Belogurov, G. A., Magretova, N. N., Baykov, A. A., and Lahti, R. (2007) Biochem. J., 408, 327-333). We herein describe a more evolved type of CBS-pyrophosphatase from Clostridium perfringens (cpCBS-PPase) that additionally contains a DRTGG domain between the two CBS domains in the regulatory part. cpCBS-PPase retained the ability of mtCBS-PPase to be inhibited by micromolar concentrations of AMP and ADP and activated by ATP and was additionally activated by diadenosine polyphosphates (AP(n)A) with n > 2. Stopped-flow measurements using a fluorescent nucleotide analog, 2'(3')-O-(N-methylanthranoyl)-AMP, revealed that cpCBS-PPase interconverts through two different conformations with transit times on the millisecond scale upon nucleotide binding. The results suggest that the presence of the DRTGG domain affords greater flexibility to the regulatory part, allowing it to more rapidly undergo conformational changes in response to binding. PMID:22348476

  18. Hemorrhagic enterocolitis and death in two felines (Panthera tigris altaica and Panthera leo) associated with Clostridium perfringens type A.

    PubMed

    Zhang, Yanlong; Hou, Zhijun; Ma, Jianzhang

    2012-06-01

    Severe hemorrhagic enterocolitis was observed in a Siberian tiger (Panthera tigris altaica) and a lion (Panthera leo). Both animals developed acute depression, anorexia, and bloody diarrhea several days before death. Small and large intestines were diffusely congested, edematous, necrotic, and filled with hemorrhagic fluid, and mesenteric lymph nodes were enlarged and congested. Pure and abundant growth of gram-positive bacilli was obtained in culture under anaerobic conditions from the livers of both felines. Identification of highly virulent Clostridium perfringens Type A was based on pathologic lesions, hemolytic patterns, morphologic structure, and polymerase chain reaction. Animal inoculation assays indicated that C. perfringens Type A played an important role in the pathogenesis of both felines. PMID:22779248

  19. The epidemiology of Clostridium perfringens type A on Ontario swine farms, with special reference to cpb2-positive isolates

    PubMed Central

    2012-01-01

    Background There is poor understanding of most aspects of Clostridium perfringens type A as a possible cause of neonatal diarrhea in piglets, and the prevalence and types of C. perfringens present on Ontario swine farms is unknown. To study the prevalence of fecal C. perfringens and selected toxin genes, 48 Ontario swine farms were visited between August 2010 and May 2011, and 354 fecal samples were collected from suckling pigs, lactating sows, weanling pigs, grower-finisher pigs, and gestating sows, as well as from manure pits. The fecal samples were cultured quantitatively, and toxin genes were detected by real-time multiplex polymerase chain reaction (PCR). Results In mixed multivariable linear analysis, log10C. perfringens in fecal samples from suckling pigs were higher than that of weanling pigs, grower-finisher pigs, and manure pit samples (P <0.05). In mixed multivariable logistic analysis, the C. perfringens isolates recovered from lactating sows (OR?=?0.069, P <0.001), gestating sows (OR?=?0.020, P <0.001), grower-finishers (OR?=?0.017, P <0.001), and manure pits (OR?=?0.11, P <0.001) were less likely to be positive for the consensus beta2 toxin gene cpb2 compared to the isolates from suckling pigs. The prevalence of cpb2 in the isolates recovered from weanlings did not differ significantly from suckling pigs. C. perfringens isolates that were positive for cpb2 were more likely to carry the atypical cpb2 gene (atyp-cpb2) (OR?=?19, P <0.001) compared to isolates that were negative for cpb2. Multivariable analysis did not identify farm factors affecting the presence of consensus cpb2 and atyp-cpb2 genes. Conclusions This study provides baseline data on the prevalence of C. perfringens and associated toxin genes in healthy pigs at different stages of production on Ontario swine farms. The study suggests that if C. perfringens type A are involved in neonatal enteritis, there may be strains with specific characteristics that cannot be identified by the existing genotyping system. PMID:22947389

  20. Abilities of the mCP Agar Method and CRENAME Alpha Toxin-Specific Real-Time PCR Assay To Detect Clostridium perfringens Spores in Drinking Water

    PubMed Central

    Maheux, Andrée F.; Bérubé, Ève; Boudreau, Dominique K.; Villéger, Romain; Cantin, Philippe; Boissinot, Maurice; Bissonnette, Luc

    2013-01-01

    We first determined the analytical specificity and ubiquity (i.e., the ability to detect all or most strains) of a Clostridium perfringens-specific real-time PCR (rtPCR) assay based on the cpa gene (cpa rtPCR) by using a bacterial strain panel composed of C. perfringens and non-C. perfringens Clostridium strains. All non-C. perfringens Clostridium strains tested negative, whereas all C. perfringens strains tested positive with the cpa rtPCR, for an analytical specificity and ubiquity of 100%. The cpa rtPCR assay was then used to confirm the identity of 116 putative C. perfringens isolates recovered after filtration of water samples and culture on mCP agar. Colonies presenting discordant results between the phenotype on mCP agar and cpa rtPCR were identified by sequencing the 16S rRNA and cpa genes. Four mCP?/rtPCR+ colonies were identified as C. perfringens, whereas 3 mCP+/rtPCR? colonies were identified as non-C. perfringens. The cpa rtPCR was negative with all 51 non-C. perfringens strains and positive with 64 of 65 C. perfringens strains. Finally, we compared mCP agar and a CRENAME (concentration and recovery of microbial particles, extraction of nucleic acids, and molecular enrichment) procedure plus cpa rtPCR (CRENAME + cpa rtPCR) for their abilities to detect C. perfringens spores in drinking water. CRENAME + cpa rtPCR detected as few as one C. perfringens CFU per 100 ml of drinking water sample in less than 5 h, whereas mCP agar took at least 25 h to deliver results. CRENAME + cpa rtPCR also allows the simultaneous and sensitive detection of Escherichia coli and C. perfringens from the same potable water sample. In itself, it could be used to assess the public health risk posed by drinking water potentially contaminated with pathogens more resistant to disinfection. PMID:24077714

  1. Directed structural modification of Clostridium perfringens enterotoxin to enhance binding to claudin-5.

    PubMed

    Protze, Jonas; Eichner, Miriam; Piontek, Anna; Dinter, Stefan; Rossa, Jan; Blecharz, Kinga Gra?yna; Vajkoczy, Peter; Piontek, Joerg; Krause, Gerd

    2015-04-01

    Clostridium perfringens enterotoxin (CPE) binds to distinct claudins (Clds), which regulate paracellular barrier functions in endo- and epithelia. The C-terminal domain (cCPE) has the potential for selective claudin modulation, since it only binds to a subset of claudins, e.g., Cld3 and Cld4 (cCPE receptors). Cld5 (non-CPE receptor) is a main constituent in tight junctions (TJ) of the blood-brain barrier. We aimed to reveal claudin recognition mechanisms of cCPE and to create a basis for a Cld5-binder. By utilizing structure-based interaction models, mutagenesis and assays of cCPE-binding to the TJ-free cell line HEK293, transfected with human Cld1 and murine Cld5, we showed how cCPE-binding to Cld1 and Cld5 is prevented by two residues in extracellular loop 2 of Cld1 (Asn(150) and Thr(153)) and Cld5 (Asp(149) and Thr(151)). Binding to Cld5 is especially attenuated by the lack of a bulky hydrophobic residue like leucine at position 151. By downsizing the binding pocket and compensating for the lack of this leucine residue, we created a novel cCPE-variant; cCPEY306W/S313H binds Cld5 with nanomolar affinity (K d 33 ± 10 nM). Finally, the effective binding to endogenously Cld5-expressing blood-brain barrier model cells (murine microvascular endothelial cEND cell line) suggests cCPEY306W/S313H as basis for Cld5-specific modulation to improve paracellular drug delivery, or to target claudin overexpressing tumors. PMID:25342221

  2. Carbohydrate Recognition by an Architecturally Complex ?-N-Acetylglucosaminidase from Clostridium perfringens

    PubMed Central

    Ficko-Blean, Elizabeth; Stuart, Christopher P.; Suits, Michael D.; Cid, Melissa; Tessier, Matthew; Woods, Robert J.; Boraston, Alisdair B.

    2012-01-01

    CpGH89 is a large multimodular enzyme produced by the human and animal pathogen Clostridium perfringens. The catalytic activity of this exo-?-d-N-acetylglucosaminidase is directed towards a rare carbohydrate motif, N-acetyl-?-d-glucosamine-?-1,4-d-galactose, which is displayed on the class III mucins deep within the gastric mucosa. In addition to the family 89 glycoside hydrolase catalytic module this enzyme has six modules that share sequence similarity to the family 32 carbohydrate-binding modules (CBM32s), suggesting the enzyme has considerable capacity to adhere to carbohydrates. Here we suggest that two of the modules, CBM32-1 and CBM32-6, are not functional as carbohydrate-binding modules (CBMs) and demonstrate that three of the CBMs, CBM32-3, CBM32-4, and CBM32-5, are indeed capable of binding carbohydrates. CBM32-3 and CBM32-4 have a novel binding specificity for N-acetyl-?-d-glucosamine-?-1,4-d-galactose, which thus complements the specificity of the catalytic module. The X-ray crystal structure of CBM32-4 in complex with this disaccharide reveals a mode of recognition that is based primarily on accommodation of the unique bent shape of this sugar. In contrast, as revealed by a series of X-ray crystal structures and quantitative binding studies, CBM32-5 displays the structural and functional features of galactose binding that is commonly associated with CBM family 32. The functional CBM32s that CpGH89 contains suggest the possibility for multivalent binding events and the partitioning of this enzyme to highly specific regions within the gastrointestinal tract. PMID:22479408

  3. Characterization of Clostridium perfringens isolates obtained from 2010 to 2012 from chickens with necrotic enteritis in Korea.

    PubMed

    Park, Ji Young; Kim, Sara; Oh, Jae Young; Kim, Hye Ryoung; Jang, Il; Lee, Hee Soo; Kwon, Yong Kuk

    2015-06-01

    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 with inflammatory exudates and characterized by mucosal necrosis. All the isolated strains were identified as toxin type A using multiplex PCR for toxin typing. We found that the rate of netB-positive strains isolated from dead chickens was significantly higher (8 of 17) than the rate among healthy chickens (2 of 50). We performed antimicrobial susceptibility test with 20 selected antimicrobial agents using the disk diffusion test and found that 30 tested strains were completely resistant to 5 antibiotics and partially resistant to 6 antibiotics whereas all the strains were susceptible to 9 antimicrobial agents. Using pulsed-field gel electrophoresis analysis, the 17 strains were divided into 13 genetic clusters showing high genetic diversity. In conclusion, C. perfringens strains isolated from Korean poultry showed a high resistance to antimicrobial drugs and high genetic diversity, suggesting that continuous monitoring is essential to prevent outbreaks of necrotic enteritis in chickens. PMID:25840962

  4. Portrait of an Enzyme, a Complete Structural Analysis of a Multimodular beta-N-Acetylglucosaminidase from Clostridium perfringens

    SciTech Connect

    Ficko-Blean, E.; Gregg, K; Adams, J; Hehemann, J; Czjzek, M; Smith, S; Boraston, A

    2009-01-01

    Common features of the extracellular carbohydrate-active virulence factors involved in host-pathogen interactions are their large sizes and modular complexities. This has made them recalcitrant to structural analysis, and therefore our understanding of the significance of modularity in these important proteins is lagging. Clostridium perfringens is a prevalent human pathogen that harbors a wide array of large, extracellular carbohydrate-active enzymes and is an excellent and relevant model system to approach this problem. Here we describe the complete structure of C. perfringens GH84C (NagJ), a 1001-amino acid multimodular homolog of the C. perfringens ?-toxin, which was determined using a combination of small angle x-ray scattering and x-ray crystallography. The resulting structure reveals unprecedented insight into how catalysis, carbohydrate-specific adherence, and the formation of molecular complexes with other enzymes via an ultra-tight protein-protein interaction are spatially coordinated in an enzyme involved in a host-pathogen interaction.

  5. Use of Calcium, Potassium, and Sodium Lactates to Control Germination and Outgrowth of Clostridium perfringens Spores during Chilling of Injected Pork

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens spore germination and outgrowth by calcium (CaL), potassium (KL) or sodium (NaL) lactate in injected pork during abusive chilling regimes was investigated. Lactates (Ca, K, or Na) were incorporated into injected pork at various concentrations (1.0, 2.0, 3.0 and...

  6. Inhibition of clostridium perfringens spore germination and outgrowth by buffered vinegar and lemon juice concentrate during chilling.....of ground turkey road containing minimal ingredients

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens spore germination and outgrowth in ground turkey roast containing minimal ingredients (salt and sugar), by buffered vinegar (MoStatin V) and a blend (buffered) of lemon juice concentrate and vinegar (MoStatin LV) was evaluated. Ground turkey roast was formulat...

  7. Inhibition of Clostridium perfringens spore germination and outgrowth by lemon juice and vinegar product in reduced NaCl roast beef

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens spore germination and outgrowth in reduced sodium roast beef by a blend of buffered lemon juice concentrate and vinegar (MoStatin LV) during abusive exponential cooling was evaluated. Roast beef containing salt (NaCl; 1, 1.5, or 2%, wt/wt), blend of sodium pyro-...

  8. Characterization of bacteriophages virulent for Clostridium perfringens and identification of phage lytic enzymes as alternatives to antibiotics for potential control of the bacterium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There has been a resurgent interest in the use of bacteriophages or their gene products to control bacterial pathogens as alternatives to currently utilized antibiotics. Clostridium perfringens is a Gram-positive, spore-forming anaerobic bacterium that plays a significant role in human food-borne di...

  9. Recombinant Attenuated Salmonella enterica Serovar Typhimurium Expressing the Carboxy-Terminal Domain of Alpha Toxin from Clostridium perfringens Induces Protective Responses against Necrotic Enteritis in Chickens

    Microsoft Academic Search

    Bereket Zekarias; Hua Mo; Roy Curtiss

    2008-01-01

    Clostridium perfringens-induced necrotic enteritis (NE) is a widespread disease in chickens that causes high mortality and reduced growth performance. Traditionally, NE was controlled by the routine application of antimicrobials in the feed, a practice that currently is unpopular. Consequently, there has been an increase in the occurrence of NE, and it has become a threat to the current objective of

  10. Characterization of bacteriophages virulent for Clostridium perfringens and identification of phage lytic enzymes as alternatives to antibiotics for potential control of the bacterium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There has been a resurgent interest in the use of bacteriophages or their gene products to control bacterial pathogens as alternatives to currently utilized antibiotics. Clostridium perfringens is a Gram-positive, spore-forming anaerobic bacterium that plays a significant role in human food-borne d...

  11. INHIBITORY EFFECTS OF ORGANIC ACID SALTS ON GROWTH OF CLOSTRIDIUM PERFRINGENS FROM SPORE INOCULA DURING CHILLING OF MARINATED GROUND TURKEY BREAST

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens germination and outgrowth by salts of organic acids such as sodium lactate, sodium acetate, buffered sodium citrate and buffered sodium citrate supplemented with sodium diacetate was evaluated during continuous chilling of ground turkey. Turkey breast meat was ...

  12. CLOSTRIDIUM PERFRINGENS TYPE A ENTEROTOXEMIA IN A CAPTIVE ADULT WHITE-TAILED DEER --On the morning of 6 June 2005, a captive adult

    E-print Network

    2006 All deer removed from the captive facility also were tested for chronic wasting disease197 CLOSTRIDIUM PERFRINGENS TYPE A ENTEROTOXEMIA IN A CAPTIVE ADULT WHITE-TAILED DEER -- On the morning of 6 June 2005, a captive adult white-tailed deer (Odocoileus virginianus) was found dead

  13. Distribution of Clostridium perfringens and Fecal Sterols in a Benthic Coastal Marine Environment Influenced by the Sewage Outfall from McMurdo Station, Antarctica

    Microsoft Academic Search

    DIANE D. EDWARDS; GORDON A. MCFETERS; M. INDIRA VENKATESAN

    1998-01-01

    The spatial distribution, movement, and impact of the untreated wastewater outfall from McMurdo Station, Antarctica, were investigated under early austral summer conditions. The benthic environment was examined to determine the distribution of Clostridium perfringens in sediment cores and the intestinal contents of native invertebrates and fish along a transect of stations. These stations extended ca. 411 m south of the

  14. The effect of artemisia annua on broiler performance, intestinal microbiota and on the course of a clostridium perfringens infection applying a necrotic enteritis disease model

    Microsoft Academic Search

    Ricarda Margarete Engberg; Kai Grevsen; Elise Ivarsen; Xavier Fretté; Lars Porskjær Christensen; Ole Højberg; Bent Borg Jensen; Nuria Canibe

    2012-01-01

    The aerial parts of the plant Artemisia annua (A. annua) contain essential oils having antimicrobial properties against Clostridium perfringens Type A, the causal agent for necrotic enteritis in broilers. In two experiments, the influence of increasing dietary concentrations of dried A. annua leaves (0, 5, 10 and 20 g\\/kg) and n-hexane extract from fresh A. annua leaves (0, 125, 250

  15. Montanide(TM) ISA 71 VG adjuvant increases protection against experimental necrotic enteritis in commercial broiler chickens following vaccination with Clostridium perfringens recombinant proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was performed to compare four Clostridium perfringens recombinant proteins as vaccine candidates using the Montanide™ ISA 71 VG adjuvant in an experimental model of necrotic enteritis. Broiler chickens were immunized with clostridial recombinant proteins with ISA 71 VG, and intestinal le...

  16. A Rare Case of Secondary Bacterial Peritonitis from Clostridium perfringens in an Adult Patient with Noncirrhotic Ascites and a Krukenberg Tumor: Report of a Case.

    PubMed

    Kelley, Scott R; Kerlakian, George M

    2011-01-01

    Secondary bacterial peritonitis, in comparison to spontaneous, presents with a surgically treatable intraabdominal source for infection such as a gastrointestinal perforation or abscess and is nearly always polymicrobial. We present a rare case of secondary bacterial peritonitis from Clostridium perfringens in an adult patient with noncirrhotic ascites and a Krukenberg tumor. PMID:21785606

  17. A poultry-intestinal isolate of Campylobacter jejuni produces a bacteriocin (CUV-3) active against a range of Gram positive bacterial pathogens including Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A newly isolated bacteriocin, CUV-3, produced by a poultry cecal isolate of Campylobacter jejuni strain CUV-3 had inhibitory activity against several Gram positive bacteria including Clostridium perfringens (38 strains), Staphylococcus aureus, Staph.epidermidis and Listeria monocytogenes. The pept...

  18. Predictive model for growth of Clostridium perfringens during cooling of cooked ground pork

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A predictive dynamic model for C. perfringens spore germination and outgrowth in cooked pork products during cooling is presented. Cooked, ground pork was inoculated with C. perfringens spores and vacuum packaged. For the isothermal experiments, all samples were incubated in a water bath stabilize...

  19. Predictive model for growth of Clostridium perfringens during cooling of cooked uncured meat and poultry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Comparison of C. perfringens spore germination and outgrowth in cooked uncured products during cooling for different meat species is presented. Cooked, uncured product was inoculated with C. perfringens spores and vacuum packaged. For the isothermal experiments, all samples were incubated in a wat...

  20. Necrotic enteritis-producing strains of Clostridium perfringens displace non-necrotic enteritis strains from the gut of chicks.

    PubMed

    Barbara, Angelique J; Trinh, Hien T; Glock, Robert D; Glenn Songer, J

    2008-01-25

    We inoculated broiler chicks with mixtures of Clostridium perfringens strains to investigate the single strain dominance observed in natural cases of necrotic enteritis (NE) [Nauerby, B., Pedersen, K., Madsen, M., 2003. Analysis by pulsed-field gel electrophoresis of the genetic diversity among Clostridium perfringens isolates from chickens. Vet. Microbiol. 94, 257-266]. Pre-inoculation bacteriologic culture of chick intestines yielded up to six pulsed-field gel electrophoresis (PFGE) types of C. perfringens. Birds developed typical NE lesions in response to administration (2x per day for 4 days) of a combined inoculum comprising one NE strain (JGS4143, PFGE pattern 8) and four non-NE strains (from piglet necrotizing enteritis, chicken normal flora, human gas gangrene, and bovine neonatal enteritis). After inoculation commenced, only the NE strain was recovered through the first post-inoculation day, in spite of intense efforts to recover pre-challenge flora strains and the other challenge strains. Thereafter, pre-inoculation and previously undetected PFGE types were found, and JGS4143 became undetectable. Birds inoculated simultaneously with five NE strains (from disease in chickens or turkeys, and including JGS4143) also developed lesions, but again only JGS4143 was recovered through the 1st day post-challenge. At that time, birds began to be repopulated with pre-challenge PFGE types. Two NE strains (JGS4143 and JGS4064) produced bacteriocins, which inhibited each other and normal flora strains (n=17), while normal flora strains inhibited neither NE strains nor each other. Thus, it appears that naturally occurring dominance of the gut by NE strains can be reproduced experimentally. Bacteriocins directed against normal flora could possibly provide the necessary advantage, although inhibition of one NE strain by another suggests that other factors may be partially or completely responsible for the dominance. PMID:17850994

  1. Effect of oxygen stress on growth and survival of Clostridium perfringens, Campylobacter jejuni, and Listeria monocytogenes under different storage conditions.

    PubMed

    Al-Qadiri, Hamzah; Sablani, Shyam S; Ovissipour, Mahmoudreza; Al-Alami, Nivin; Govindan, Byju; Rasco, Barbara

    2015-04-01

    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. perfringens could grow in beef at 22 °C, with an increase of approximately 5 log under anoxic conditions and a 1-log increase under microoxic conditions. However, C. perfringens could not survive in beef held at 7 °C under microoxic and oxic storage conditions after 14 days. In an anoxic environment, C. perfringens survived in beef samples held at 7 °C, with a 1-log reduction. A cell decline was observed at 2 log under these conditions, with no surviving cells at the 1-log level. However, the results show that C. jejuni under microoxic conditions survived with declining cell numbers. Significant increases in L. monocytogenes (5 to 7 log) were observed in beef held at 22 °C for 5 days, with the lowest levels recovered under anoxic conditions. L. monocytogenes in refrigerated storage increased by a factor of 2 to 4 log. It showed the greatest growth under oxic conditions, with significant growth under anoxic conditions. These findings can be used to enhance food safety in vacuum-packed and modified atmosphere-packaged food products. PMID:25836393

  2. A Sporulation Factor Is Involved in the Morphological Change of Clostridium perfringens Biofilms in Response to Temperature

    PubMed Central

    Obana, Nozomu; Nakamura, Kouji

    2014-01-01

    Biofilm formation has been associated with bacterial pathogenesis, such as nosocomial and chronic infections, as the resistance of biofilms to environmental stresses has increased. Clostridium perfringens is a Gram-positive spore-forming anaerobic pathogen. This organism survives antibiotic treatment through the formation of biofilms or spores, but the environmental and regulatory factors involved in the biofilm formation remain unclear. Here, we observed that temperature regulates C. perfringens biofilm morphology. At 37°C, C. perfringens adhered to the substrate surface and formed a flat, thin biofilm, herein referred to as adhered biofilm. However, at 25°C, this bacterium did not adhere and produced a threadlike extracellular matrix, forming a viscous, thick biofilm, herein referred to as pellicle biofilm. Pellicle biofilm formation requires the sporulation master regulator, Spo0A, and the toxin regulator, CtrAB, and is enhanced in the absence of the global repressor, AbrB. These transcriptional regulator genes are regulated by each other and temperature. Adhered-biofilm formation requires AbrB and pilA2, which encodes a component of type IV pili (TFP). TFP expression was activated at 37°C and regulated through Spo0A, AbrB, and CtrAB. These results indicate that the morphology of C. perfringens biofilm is dependent on temperature through the differential production of extracellular matrix and the activity of TFP. Moreover, pellicle biofilm formation is involved in sporulation and toxin production. Here, we demonstrated that clostridial biofilm formation is closely associated with sporulation and that the morphological change of the biofilms could play an important role in the pathogenesis of this organism. PMID:24509316

  3. Control of Clostridium perfringens germination and outgrowth by buffered sodium citrate during chilling of roast beef and injected pork.

    PubMed

    Thippareddi, H; Juneja, V K; Phebus, R K; Marsden, J L; Kastner, C L

    2003-03-01

    Inhibition of the germination and outgrowth of Clostridium perfringens by buffered sodium citrate (Ional) and buffered sodium citrate supplemented with sodium diacetate (Ional Plus) during the abusive chilling of roast beef and injected pork was evaluated. Beef top rounds or pork loins were injected with a brine containing NaCl, potato starch, and potassium tetrapyrophosphate to yield final in-product concentrations of 0.85, 0.25, and 0.20%, respectively. Products were ground and mixed with Ional or Ional Plus at 0, 0.5, 1.0, and 2.0%. Each product was mixed with a three-strain C. perfringens spore cocktail to obtain final spore concentrations of ca. 2.5 log10 spores per g. Chilling of roast beef from 54.4 to 7.2 degrees C resulted in C. perfringens population increases of 1.51 and 5.27 log10 CFU/g for 18- and 21-h exponential chill rates, respectively, while chilling of injected pork resulted in increases of 3.70 and 4.41 log10 CFU/g. The incorporation of Ional into the roast beef formulation resulted in C. perfringens population reductions of 0.98, 1.87, and 2.47 log10 CFU/g with 0.5, 1.0, and 2.0% Ional, respectively, over 18 h of chilling, while > or = 1.0% Ional Plus was required to achieve similar reductions (reductions of 0.91 and 2.07 log10 CFU/g were obtained with 1.0 and 2.0% Ional Plus, respectively). An Ional or Ional Plus concentration of > or = 1.0% was required to reduce C. perfringens populations in roast beef or injected pork chilled from 54.4 to 7.2 degrees C in 21 h. Cooling times for roast beef or injected pork products after heat processing can be extended to 21 h through the incorporation of > or = 1.0% Ional or Ional Plus into the formulation to reduce the potential risk of C. perfringens germination and outgrowth. PMID:12636288

  4. Clostridium perfringens challenge and dietary fat type affect broiler chicken performance and fermentation in the gastrointestinal tract.

    PubMed

    Józefiak, D; Kiero?czyk, B; Rawski, M; Hejdysz, M; Rutkowski, A; Engberg, R M; Højberg, O

    2014-06-01

    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 Experiment 1, birds were fed diets containing soybean oil, palm kernel fatty acid distillers, rendered pork fat and lard. In Experiment 2, birds were fed diets containing rapeseed oil, coconut oil, beef tallow and palm oil. In both experiments, the birds were either not challenged or challenged with a mixture of three C. perfringens type A strains. Irrespective of the fat type present in the diet, C. perfringens did not affect broiler chicken body weight gain (BWG) and mortality in either of the two experiments. The BWG was affected by dietary fat type in both experiments, indicating that the fatty acid composition of the fat source affects broiler growth performance. In particular, the inclusion of animal fats tended to improve final BW to a greater extent compared with the inclusion of unsaturated vegetable oils. In Experiment 2, irrespective of the dietary fat type present in the diet, C. perfringens challenge significantly impaired feed conversion ratio in the period from 14 to 28 days (1.63 v. 1.69) and at 42 days (1.65 v. 1.68). In both experiments apparent metabolizable energy values were affected by dietary fat type. Irrespective of the fat type present in the diet, C. perfringens challenge decreased the digesta pH in the crop and ileum, but had no effect in cecal contents. Moreover, in Experiment 1, total organic acid concentration in the ileum was two to three times lower on soybean oil diets as compared with other treatments, indicating that C. perfringens as well as dietary fat type significantly affects microbiota activity in the broiler chicken gastrointestinal tract. PMID:24674938

  5. A Wide Variety of Clostridium perfringens Type A Food-Borne Isolates That Carry a Chromosomal cpe Gene Belong to One Multilocus Sequence Typing Cluster

    PubMed Central

    Xiao, Yinghua; Wagendorp, Arjen; Moezelaar, Roy; Abee, Tjakko

    2012-01-01

    Of 98 suspected food-borne Clostridium perfringens isolates obtained from a nationwide survey by the Food and Consumer Product Safety Authority in The Netherlands, 59 strains were identified as C. perfringens type A. Using PCR-based techniques, the cpe gene encoding enterotoxin was detected in eight isolates, showing a chromosomal location for seven isolates and a plasmid location for one isolate. Further characterization of these strains by using (GTG)5 fingerprint repetitive sequence-based PCR analysis distinguished C. perfringens from other sulfite-reducing clostridia but did not allow for differentiation between various types of C. perfringens strains. To characterize the C. perfringens strains further, multilocus sequence typing (MLST) analysis was performed on eight housekeeping genes of both enterotoxic and non-cpe isolates, and the data were combined with a previous global survey covering strains associated with food poisoning, gas gangrene, and isolates from food or healthy individuals. This revealed that the chromosomal cpe strains (food strains and isolates from food poisoning cases) belong to a distinct cluster that is significantly distant from all the other cpe plasmid-carrying and cpe-negative strains. These results suggest that different groups of C. perfringens have undergone niche specialization and that a distinct group of food isolates has specific core genome sequences. Such findings have epidemiological and evolutionary significance. Better understanding of the origin and reservoir of enterotoxic C. perfringens may allow for improved control of this organism in foods. PMID:22865060

  6. Use of site-directed mutagenesis to probe structure-function relationships of alpha-toxin from Clostridium perfringens.

    PubMed Central

    Guillouard, I; Garnier, T; Cole, S T

    1996-01-01

    The NH2-terminal domain of the alpha-toxin of Clostridium perfringens is highly homologous to the complete phospholipase C from Bacillus cereus (PC-PLC), for which a high-resolution crystal structure is available. This structural information was used as the basis of a site-directed mutagenesis strategy in which critical amino acid residues of alpha-toxin involved in zinc binding, interaction with substrate, or catalysis were replaced. Biochemical studies with the corresponding toxin variants indicate that there is probably a single active site endowed with lecithinase, sphingomyelinase, and hemolytic activities. By using a highly purified variant in which the catalytic aspartate residue at position 56 was replaced by asparagine, it was shown that phospholipase activity was essential for lethality in vivo and for mediating platelet aggregation in vitro. PMID:8698464

  7. Susceptibility of anaerobic bacteria to nine antimicrobial agents and demonstration of decreased susceptibility of Clostridium perfringens to penicillin.

    PubMed

    Marrie, T J; Haldane, E V; Swantee, C A; Kerr, E A

    1981-01-01

    The activity of moxalactam, cefoxitin, cephalothin, cefamandole, chloramphenicol, clindamycin, metronidazole, and ticarcillin was determined against 344 isolates of anaerobic bacteria. The activity of penicillin G was determined as well for 234 isolates not of the Bacteroides fragilis group. Moxalactam was more active than cephalothin and cefamandole and slightly less active than cefoxitin. Metronidazole was the most active antimicrobial agent against the B. fragilis group, whereas chloramphenicol was most active overall. Clostridium species were the most resistant group of organisms tested. Relatively high concentrations of penicillin were required to inhibit the C. perfringens strains: 80% at 0.5 U/ml and 100% at 16 U/ml. Our study demonstrates the need for periodic anaerobe susceptibility testing in order to better guide empiric antibiotic therapy. PMID:6264842

  8. The Sialidases of Clostridium perfringens Type D Strain CN3718 Differ in Their Properties and Sensitivities to Inhibitors

    PubMed Central

    Li, Jihong

    2014-01-01

    Clostridium perfringens causes histotoxic infections and diseases originating in animal or human intestines. A prolific toxin producer, this bacterium also produces numerous enzymes, including sialidases, that may facilitate infection. C. perfringens type D strain CN3718 carries genes encoding three sialidases, including two large secreted sialidases (named NanI and NanJ) and one small sialidase (named NanH) that has an intracellular location in log-phase cultures but is present in supernatants of death phase cultures. Using isogenic mutants of CN3718 that are capable of expressing only NanJ, NanI, or NanH, the current study characterized the properties and activities of each sialidase. The optimal temperature determined for NanJ or NanH enzymatic activity was 37°C or 43°C, respectively, while NanI activity increased until temperature reached 48°C. NanI activity was also the most resistant against higher temperatures. All three sialidases showed optimal activities at pH 5.5. Compared to NanJ or NanH, NanI contributed most to the sialidase activity in CN3718 culture supernatants, regardless of the substrate sialic acid linkage; NanI also released the most sialic acid from Caco-2 cells. Only NanI activity was enhanced by trypsin pretreatment and then only for substrates with an ?-2,3- or ?-2,6-sialic acid linkage. NanJ and NanI activities were more sensitive than NanH activity to two sialidase inhibitors (N-acetyl-2,3-dehydro-2-deoxyneuraminic acid and siastatin B). The activities of the three sialidases were affected differently by several metal ions. These results indicated that each C. perfringens sialidase has distinct properties, which may allow these enzymes to play different roles depending upon environmental conditions. PMID:24375134

  9. Global Phenotypic Characterization of Effects of Fluoroquinolone Resistance Selection on the Metabolic Activities and Drug Susceptibilities of Clostridium perfringens Strains

    PubMed Central

    Park, Miseon

    2014-01-01

    Fluoroquinolone resistance affects toxin production of Clostridium perfringens strains differently. To investigate the effect of fluoroquinolone resistance selection on global changes in metabolic activities and drug susceptibilities, four C. perfringens strains and their norfloxacin-, ciprofloxacin-, and gatifloxacin-resistant mutants were compared in nearly 2000 assays, using phenotype microarray plates. Variations among mutant strains resulting from resistance selection were observed in all aspects of metabolism. Carbon utilization, pH range, osmotic tolerance, and chemical sensitivity of resistant strains were affected differently in the resistant mutants depending on both the bacterial genotype and the fluoroquinolone to which the bacterium was resistant. The susceptibilities to gentamicin and erythromycin of all resistant mutants except one increased, but some resistant strains were less susceptible to amoxicillin, cefoxitin, ceftriaxone, chloramphenicol, and metronidazole than their wild types. Sensitivity to ethidium bromide decreased in some resistant mutants and increased in others. Microarray analysis of two gatifloxacin-resistant mutants showed changes in metabolic activities that were correlated with altered expression of various genes. Both the chemical structures of fluoroquinolones and the genomic makeup of the wild types influenced the changes found in resistant mutants, which may explain some inconsistent reports of the effects of therapeutic use of fluoroquinolones on clinical isolates of bacteria. PMID:25587280

  10. Hypermotility in Clostridium perfringens strain SM101 is due to spontaneous mutations in genes linked to cell division.

    PubMed

    Liu, Hualan; McCord, Kristin D; Howarth, Jonathon; Popham, David L; Jensen, Roderick V; Melville, Stephen B

    2014-07-01

    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 suggests that they form long, thin filaments that move rapidly away from a colony, analogously to swarmer cells in bacteria with flagella. To identify the cause(s) of the hypermotility phenotype, the genome sequences of normal strains and their direct hypermotile derivatives were determined and compared. Strains SM124 and SM127, hypermotile derivatives of strains SM101 and SM102, respectively, contained 10 and 6 single nucleotide polymorphisms (SNPs) relative to their parent strains. While SNPs were located in different genes in the two sets of strains, one feature in common was mutations in cell division genes, an ftsI homolog in strain SM124 (CPR_1831) and a minE homolog in strain SM127 (CPR_2104). Complementation of these mutations with wild-type copies of each gene restored the normal motility phenotype. A model explaining the principles underlying the hypermotility phenotype is presented. PMID:24748614

  11. Antimicrobial activity of Satureja montana L. essential oil against Clostridium perfringens type A inoculated in mortadella -type sausages formulated with different levels of sodium nitrite

    Microsoft Academic Search

    Thales Leandro Coutinho de Oliveira; Rodrigo de Araújo Soares; Eduardo Mendes Ramos; Maria das Graças Cardoso; Eduardo Alves; Roberta Hilsdorf Piccoli

    2011-01-01

    This research evaluated the antimicrobial effect of the winter savory (Satureja montana L.) essential oil (EO) against Clostridium perfringens type A (ATCC 3624) inoculated in mortadella-type sausages formulated with different levels of sodium nitrite (NaNO2: 0ppm, 100ppm and 200ppm) in addition to EO at concentrations of 0.0%, 0.78%, 1.56% and 3.125% stored at 25°C for 30days. The EO extracted by

  12. PREDICTIVE MODEL FOR GROWTH OF CLOSTRIDIUM PERFRINGENS DURING COOLING OF COOKED UNCURED BEEF

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper considers two models that have been used for modeling growth of C. perfringens during cooling. Using a common approach or methodology for constructing models, there was no appreciable difference between the model predictions when the population of cells was within the lag or exponential ...

  13. Growth potential of Clostridium perfringens from spores in acidified beef, pork and poultry products during chilling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ability of C. perfringens to germinate and grow in acidified ground beef as well as in ten commercially prepared acidified beef, pork and poultry products was assessed. The pH of ground beef was adjusted using organic vinegar to achieve various pH values between 5.0 and 5.6; the pH of the commer...

  14. NUMERICAL ANALYSIS OF THE GROWTH OF CLOSTRIDIUM PERFRINGENS IN COOKED BEEF UNDER ISOTHERMAL AND DYNAMIC CONDITIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to develop a mathematical methodology to estimate the growth of C. perfringens in cooked beef under dynamic temperature conditions. Two differential equations governing the lag phase development and cell multiplication were proposed and solved using a 4th-order Runge...

  15. Enhancing Chicken Mucosal IgA Response Against Clostridium Perfringens a-toxin 

    E-print Network

    Chen, Chang-Hsin 1977-

    2012-07-27

    Necrotic enteritis (NE) is an economically important enteric disease of broiler chicken primarily caused by a-toxin (Cpa) secreted by C. perfringens type A. Mice immunized with recombinant C-terminal domain of Cpa (CpaCD) had transient and fewer...

  16. Dynamic determination of kinetic parameters and computer simulation of growth of Clostridium perfringens in cooked beef

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this research was to develop a new one-step methodology that uses a dynamic approach to directly construct a tertiary model for prediction of the growth of C. perfringens in cooked beef. This methodology was based on numerical analysis and optimization of both primary and secondary...

  17. Effect of rooibos (Aspalathus linearis) on growth control of Clostridium perfringens and lipid oxidation of ready-to-eat Jokbal (pig's trotters).

    PubMed

    Park, Hee Jin; Park, Keun-Cheol; Yoon, Ki Sun

    2014-12-01

    This study investigated the antimicrobial effects of rooibos (tea extract), potassium lactate (PL) and sodium diacetate (SDA) mixture alone or in combinations on the growth of Clostridium perfringens vegetative cell and spore in ready-to-eat (RTE) Jokbal (pig's trotters). Addition of a combination of 10% rooibos and 4% PL + SDA inhibit growth of C. perfringens vegetative cell in Jokbal at 24 °C and 36 °C. The significant inhibition on germination and growth of C. perfringens spores was also observed in Jokbal with a combination of 10% rooibos and 4% PL + SDA (PL: 2.24%, SDA: 0.16%) at 24 °C. The Jokbal treated with 10% rooibos and 4% PL + SDA mixture had significantly (P < 0.05) lower TBARS values than the control at 10 and 24 °C. The lipid oxidation inhibition effect was the highest (P < 0.05) in anaerobic packed Jokbal with 10% rooibos. The addition of a combination of 10% rooibos and 4% PL + SDA during the processing of Jokbal prevented the growth of C. perfringens and the germination and growth of C. perfringens spores at room temperature. This study shows rooibos tea as a valuable natural food preservative in meat products. PMID:25394229

  18. Identification and characterization of Clostridium perfringens beta toxin variants with differing trypsin sensitivity and in vitro cytotoxicity activity.

    PubMed

    Theoret, James R; Uzal, Francisco A; McClane, Bruce A

    2015-04-01

    By producing toxins, Clostridium perfringens causes devastating diseases of both humans and animals. C. perfringens beta toxin (CPB) is the major virulence determinant for type C infections and is also implicated in type B infections, but little is known about the CPB structure-function relationship. Amino acid sequence comparisons of the CPBs made by 8 randomly selected isolates identified two natural variant toxins with four conserved amino acid changes, including a switch of E to K at position 168 (E168K) that introduces a potential trypsin cleavage site into the CPB protein of strain JGS1076. To investigate whether this potential trypsin cleavage site affects sensitivity to trypsin, a primary host defense against this toxin, the two CPB variants were assayed for their trypsin sensitivity. The results demonstrated a significant difference in trypsin sensitivity, which was linked to the E168K switch by using site-directed recombinant CPB (rCPB) mutants. The natural CPB variants also displayed significant differences in their cytotoxicity to human endothelial cells. This cytotoxicity difference was mainly attributable to increased host cell binding rather than the ability to oligomerize or form functional pores. Using rCPB site-directed mutants, differences in cytotoxicity and host cell binding were linked to an A300V amino acid substitution in the strain JGS1076 CPB variant that possessed more cytotoxic activity. Mapping of sequence variations on a CPB structure modeled using related toxins suggests that the E168K substitution is surface localized and so can interact with trypsin and that the A300V substitution is located in a putative binding domain of the CPB toxin. PMID:25643999

  19. Novel Clostridium perfringens enterotoxin suicide gene therapy for selective treatment of claudin-3- and -4-overexpressing tumors.

    PubMed

    Walther, W; Petkov, S; Kuvardina, O N; Aumann, J; Kobelt, D; Fichtner, I; Lemm, M; Piontek, J; Blasig, I E; Stein, U; Schlag, P M

    2012-05-01

    Bacterial toxins are known to be effective for cancer therapy. Clostridium perfringens enterotoxin (CPE) is produced by the bacterial Clostridium type A strain. The transmembrane proteins claudin-3 and -4, often overexpressed in numerous human epithelial tumors (for example, colon, breast, pancreas, prostate and ovarian), are the targeted receptors for CPE. CPE binding to them triggers formation of membrane pore complexes leading to rapid cell death. In this study, we aimed at selective tumor cell killing by CPE gene transfer. We generated expression vectors bearing the bacterial wild-type CPE cDNA (wtCPE) or translation-optimized CPE (optCPE) cDNA for in vitro and in vivo gene therapy of claudin-3- and -4-overexpressing tumors. The CPE expression analysis at messenger RNA and protein level revealed more efficient expression of optCPE compared with wtCPE. Expression of optCPE showed rapid cytotoxic activity, hightened by CPE release as bystander effect. Cytotoxicity of up to 100% was observed 72?h after gene transfer and is restricted to claudin-3-and -4-expressing tumor lines. MCF-7 and HCT116 cells with high claudin-4 expression showed dramatic sensitivity toward CPE toxicity. The claudin-negative melanoma line SKMel-5, however, was insensitive toward CPE gene transfer. The non-viral intratumoral in vivo gene transfer of optCPE led to reduced tumor growth in MCF-7 and HCT116 tumor-bearing mice compared with the vector-transfected control groups. This novel approach demonstrates that CPE gene transfer can be employed for a targeted suicide gene therapy of claudin-3- and -4-overexpressing tumors, leading to the rapid and efficient tumor cell killing in vitro and in vivo. PMID:21975465

  20. Location and stoichiometry of the protease CspB and the cortex-lytic enzyme SleC in Clostridium perfringens spores.

    PubMed

    Banawas, Saeed; Korza, George; Paredes-Sabja, Daniel; Li, Yunfeng; Hao, Bing; Setlow, Peter; Sarker, Mahfuzur R

    2015-09-01

    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 both germination and apparent viability of C. perfringens spores in the absence of an exogenous lytic enzyme. In addition, quantitative Western blot analyses showed that there are approximately 2000 and 130,000 molecules of CspB and pro-SleC, respectively, per C. perfringens SM101 spore, consistent with CspB's role in acting catalytically on pro-SleC to convert this zymogen to the active enzyme. PMID:25998819

  1. Clostridium perfringens Phospholipase C Induced ROS Production and Cytotoxicity Require PKC, MEK1 and NF?B Activation

    PubMed Central

    Monturiol-Gross, Laura; Flores-Díaz, Marietta; Pineda-Padilla, Maria Jose; Castro-Castro, Ana Cristina; Alape-Giron, Alberto

    2014-01-01

    Clostridium perfringens phospholipase C (CpPLC), also called ?-toxin, is the most toxic extracellular enzyme produced by this bacteria and is essential for virulence in gas gangrene. At lytic concentrations, CpPLC causes membrane disruption, whereas at sublytic concentrations this toxin causes oxidative stress and activates the MEK/ERK pathway, which contributes to its cytotoxic and myotoxic effects. In the present work, the role of PKC, ERK 1/2 and NF?B signalling pathways in ROS generation induced by CpPLC and their contribution to CpPLC-induced cytotoxicity was evaluated. The results demonstrate that CpPLC induces ROS production through PKC, MEK/ERK and NF?B pathways, the latter being activated by the MEK/ERK signalling cascade. Inhibition of either of these signalling pathways prevents CpPLC's cytotoxic effect. In addition, it was demonstrated that NF?B inhibition leads to a significant reduction in the myotoxicity induced by intramuscular injection of CpPLC in mice. Understanding the role of these signalling pathways could lead towards developing rational therapeutic strategies aimed to reduce cell death during a clostridialmyonecrosis. PMID:24466113

  2. Antagonism exerted by an association of a Bacteroides thetaiotaomicron strain and a Fusobacterium necrogenes strain against Clostridium perfringens in gnotobiotic mice and in fecal suspensions incubated in vitro.

    PubMed Central

    Yurdusev, N; Ladire, M; Ducluzeau, R; Raibaud, P

    1989-01-01

    Antagonism between an association of Bacteroides thetaiotaomicron and Fusobacterium necrogenes strains and two strains of Clostridium perfringens was evidenced both in vivo in gnotobiotic mice and ex vivo in fecal suspensions incubated for 22 h at 37 degrees C. Several features of this antagonism were similar in and ex vivo. (i) An obligate and continuous synergy between B. thetaiotaomicron and F. necrogenes was required; (ii) the two C. perfringens strains did not respond to the same extent to this antagonism; and (iii) expression of the antagonism was host and diet dependent. Neither diffusible nor soluble inhibitory substances were detectable in feces of gnotobiotic mice, nor could depletion of nutrients be identified as causing antagonism in both in and ex vivo experiments. Our findings support the hypothesis that a reversible bacteriostasis induced by the inhibitory strains acting together continuously, and hindering the target strain from utilizing available nutrients, was responsible for this antagonism. PMID:2537255

  3. Inhibitory effects of organic acid salts on growth of Clostridium perfringens from spore inocula during chilling of marinated ground turkey breast.

    PubMed

    Juneja, V K; Thippareddi, H

    2004-06-01

    Inhibition of Clostridium perfringens germination and outgrowth by salts of organic acids such as sodium lactate, sodium acetate, buffered sodium citrate and buffered sodium citrate supplemented with sodium diacetate was evaluated during continuous chilling of ground turkey. Turkey breast meat was injected with a brine-containing NaCl, potato starch and potassium tetra pyrophosphate to yield final in-product concentrations of 0.85%, 0.25% and 0.20%, respectively. The meat was ground, mixed with either sodium lactate (1%, 2%, 3% or 4%), sodium acetate (1% or 2%), buffered sodium citrate (Ional, 1%) or buffered sodium citrate supplemented with sodium diacetate (Ional Plus trade mark, 1%), in addition to a control that did not contain added antimicrobials. Each product was mixed with a three-strain C. perfringens spore cocktail to obtain final spore concentrations of ca. 2.8 log10 spores/g. Inoculated products (10 g) were packaged into cook-in-bags (2 x 3 in.), vacuum sealed, cooked at 60 degrees C for 1 h, and subsequently chilled from 54.4 to 7.2 degrees C in 15, 18 and 21 h following exponential chilling rates. Products were sampled immediately after cooking and then after chilling. Chilling of cooked turkey following 15, 18 and 21 h chill rates resulted in germination and outgrowth of C. perfringens spores to 6.6, 7.58 and 7.95 log10 CFU/g populations, respectively, from initial spore populations of ca. 2.80 log10 CFU/g. Incorporation of sodium lactate (1%), sodium acetate (1%), Ional or Ional Plus (1%) substantially inhibited germination and outgrowth of C. perfringens spores compared to controls. Final C. perfringens total populations of 3.12, 3.10, 2.38 and 2.92 log10 CFU/g, respectively, were observed following a 15-h exponential chill rate. Similar inhibitory effects were observed for 18 and 21 chill rates with the antimicrobials at 1% concentrations. While sodium lactate and sodium acetate concentrations of 1% were sufficient to control C. perfringens germination and outgrowth (<1.0 log10 CFU/g growth) following 15 h chill rates, higher concentrations were required for 18 and 21 h chill rates. Ional at 1% concentration was effective in inhibiting germination and outgrowth to <1.0 log10 CFU/g of C. perfringens for all three chill rates (15, 18 and 21 h) tested. Use of sodium salts of organic acids in formulation of ready-to-eat meat products can reduce the risk of C. perfringens spore germination and outgrowth during chilling. PMID:15135954

  4. Phenotypic Characterization of Enterotoxigenic Clostridium perfringens Isolates from Non-foodborne Human Gastrointestinal Diseases

    Microsoft Academic Search

    Renee E Collie; John F Kokai-Kun; Bruce A McClane

    1998-01-01

    Clostridium perfringensenterotoxin (CPE) has been implicated as an important virulence factor inC. perfringenstype A food poisoning and several non-foodborne human gastrointestinal (GI) illnesses, including antibiotic-associated diarrhea (AAD) and sporadic diarrhea (SPOR). Recent studies have revealed genotypic differences betweencpe-positive isolates originating from different disease sources, with most, or all, food poisoning isolates carrying a chomosomalcpeand most, or all, non-foodborne human GI

  5. 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

    PubMed Central

    Xiao, Yinghua; van Hijum, Sacha A. F. T.; Abee, Tjakko; Wells-Bennik, Marjon H. J.

    2015-01-01

    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, biomass accumulation (OD600), the total viable counts of cells plus spores, the viable count of heat resistant spores alone, the pH of the supernatant, enterotoxin production and dipicolinic acid accumulation. Subsequently, whole-genome expression profiling during key phases of the sporulation process was performed using DNA microarrays, and genes were clustered based on their time-course expression profiles during sporulation. The majority of previously characterized C. perfringens germination genes showed upregulated expression profiles in time during sporulation and belonged to two main clusters of genes. These clusters with up-regulated genes contained a large number of C. perfringens genes which are homologs of Bacillus genes with roles in sporulation and germination; this study therefore suggests that those homologs are functional in C. perfringens. A comprehensive homology search revealed that approximately half of the upregulated genes in the two clusters are conserved within a broad range of sporeforming Firmicutes. Another 30% of upregulated genes in the two clusters were found only in Clostridium species, while the remaining 20% appeared to be specific for C. perfringens. These newly identified genes may add to the repertoire of genes with roles in sporulation and determining spore properties including germination behavior. Their exact roles remain to be elucidated in future studies. PMID:25978838

  6. Use of organic acids for the control of Clostridium perfringens in cooked vacuum-packaged restructured roast beef during an alternative cooling procedure.

    PubMed

    Sabah, J R; Thippareddi, H; Marsden, J L; Fung, D Y C

    2003-08-01

    This study was conducted to determine how well Clostridium perfringens spores germinate and grow in restructured roast beef treated with different commercial organic salts during an alternative chilling procedure. The meat was prepared according to an industrial recipe (10% water, 1.5% sodium chloride, and 0.5% sodium triphosphate). The base meat was treated with sodium citrate at 2 or 4.8% (wt/wt), buffered to a pH of 5.6, 5.0, or 4.4 (six treatments); a 60% (wt/wt) solution of sodium lactate at 2 or 4.8% (wt/wt); sodium acetate at 0.25% (wt/wt); or sodium diacetate at 0.25% (wt/wt). Untreated meat was used as a control. Meat samples were inoculated with a three-strain cocktail of C. perfringens spores (strains ATCC 10388, NCTC 8238, and NCTC 8239). Meat was vacuum packaged in bags and cooked in a stirred water bath to an internal temperature of 75 degrees C for 20 min, and then the bags were cooled from 54.4 to 4.4 degrees C within 18 h. Samples were taken after inoculation, after cooking, and after chilling. Spore and vegetative cell counts were obtained after incubation at 37 degrees C for 8 to 10 h in Fung's Double Tubes containing tryptose sulfite agar without egg yolk enrichment. Cooking was not sufficient to eliminate C. perfringens spores. Over the 18-h cooling period, sodium citrate, sodium lactate, and sodium diacetate reduced the growth of C. perfringens to < 1 log unit, a growth level that meets U.S. Department of Agriculture performance standards. The use of sodium citrate or sodium lactate at a concentration of > or = 2% (wt/wt) inhibited C. perfringens growth over the 18-h cooling period. PMID:12929827

  7. A Fatal Spontaneous Gas Gangrene due to Clostridium perfringens during Neutropenia of Allogeneic Stem Cell Transplantation: Case Report and Literature Review

    PubMed Central

    Lee, Hae-Lim; Cho, Sung-Yeon; Ko, Yumi; Hyun, Ji In; Kim, Bo Kyoung; Seo, Jae Hyun; Lee, Jung Woo; Lee, Seok

    2014-01-01

    Most cases of gas gangrene caused by Clostridium species begin with trauma-related injuries but in rare cases, spontaneous gas gangrene (SGG) can occur when patients have conditions such as advanced malignancy, diabetes, or immunosuppression. Clostridium perfringens, a rare cause of SGG, exists as normal flora of skin and intestines of human. Adequate antibiotics with surgical debridement of infected tissue is the only curative therapeutic management. Mortality rate among adults is reported range of 67-100% and majority of deaths are occurred within 24 hours of onset. We experienced a case of SGG on the trunk, buttock and thigh in a neutropenic patient with acute lymphoblastic leukemia. His clinical course was rapid and fatal during pre-engraftment neutropenic period of allogeneic stem cell transplantation. PMID:25298910

  8. Clostridium perfringens bacteriophages ?CP39O and ?CP26F: genomic organization and proteomic analysis of the virions

    PubMed Central

    Fouts, Derrick E.; Simmons, Mustafa; Garrish, Johnna K.; Kuntz, Robin L.; Woolsey, Rebekah; Schegg, Kathleen M.; Kropinski, Andrew M.; Ackermann, Hans-W.; Siragusa, Gregory R.

    2014-01-01

    Poultry intestinal material, sewage and poultry processing drainage water were screened for virulent Clostridium perfringens bacteriophages. Viruses isolated from broiler chicken offal washes (O) and poultry feces (F), designated ?CP39O and ?CP26F, respectively, produced clear plaques on host strains. Both bacteriophages had isometric heads of 57 nm in diameter with 100-nm non-contractile tails characteristic of members of the family Siphoviridae in the order Caudovirales. The double-strand DNA genome of bacteriophage ?CP39O was 38,753 base pairs (bp), while the ?CP26F genome was 39,188 bp, with an average GC content of 30.3%. Both viral genomes contained 62 potential open reading frames (ORFs) predicted to be encoded on one strand. Among the ORFs, 29 predicted proteins had no known similarity while others encoded putative bacteriophage capsid components such as a pre-neck/appendage, tail, tape measure and portal proteins. Other genes encoded a predicted DNA primase, single-strand DNA-binding protein, terminase, thymidylate synthase and a transcription factor. Potential lytic enzymes such as a fibronectin-binding autolysin, an amidase/hydrolase and a holin were encoded in the viral genomes. Several ORFs encoded proteins that gave BLASTP matches with proteins from Clostridium spp. and other Gram-positive bacterial and bacteriophage genomes as well as unknown putative Collinsella aerofaciens proteins. Proteomics analysis of the purified viruses resulted in the identification of the putative pre-neck/appendage protein and a minor structural protein encoded by large open reading frames. Variants of the portal protein were identified, and several mycobacteriophage gp6-like protein variants were detected in large amounts relative to other virion proteins. The predicted amino acid sequences of the pre-neck/appendage proteins had major differences in the central portion of the protein between the two phage gene products. Based on phylogenetic analysis of the large terminase protein, these phages are predicted to be pac-type, using a head-full DNA packaging strategy. PMID:20963614

  9. C-Terminus of Clostridium perfringens Enterotoxin Downregulates CLDN4 and Sensitizes Ovarian Cancer Cells to Taxol and Carboplatin

    PubMed Central

    Gao, Zhijian; Xu, Xiaoyin; McClane, Bruce; Zeng, Qing; Litkouhi, Babak; Welch, William R.; Berkowitz, Ross S.; Mok, Samuel C.; Garner, Elizabeth I.O.

    2010-01-01

    Purpose We have previously demonstrated that CLDN4 (encoding claudin-4), a cell tight junction (TJ) protein, is highly expressed in human epithelial ovarian cancers (EOC) but undetectable in normal ovaries. CLDN4 has been identified as a specific receptor for c-terminus of Clostridium perfringens enterotoxin (C-CPE), a nontoxic molecule that may disrupt TJ barrier function and enhance cellular absorption. The purpose of this study was to determine the potential clinical applications of C-CPE and its effects on CLDN4 expression in EOC. Experimental design Using a 3D culture model and monolayer culture of EOC cells, we examined the effects of C-CPE on CLDN4 expression by qRT-PCR, immunoflorescence and Western blot. The synergistic effect of C-CPE to clinically relevant chemotherapies (Taxol and Carboplatin) was observed in EOC culture and xenograft mice. Furthermore, we determined through oligonucleotide microarray analysis the transcript profile alterations dysregulated as a consequence of C-CPE treatment. Results C-CPE treatment decreased protein expression and relocated CLDN4 from cell-cell contact regions to the cytoplasm. Particularly, C-CPE sensitized EOC cells to chemotherapeutic administration at low dosages and significantly inhibited tumor growth in a non-toxic manner. Furthermore, we provided genome-wide molecular evidence that C-CPE treatment is involved in the stimulation of the ubiquitin-proteasome pathway and the inhibition of cell metabolism in EOC cells. Conclusions The addition of C-CPE can enhance the effectiveness of Taxol or Carboplatin and significantly inhibited EOC cell growth in a CLDN4-dependent, suggesting that C-CPE may have promising therapeutic potential for EOC. PMID:21123456

  10. Removal and fate of Cryptosporidium parvum, Clostridium perfringens and small-sized centric diatoms (Stephanodiscus hantzschii) in slow sand filters.

    PubMed

    Hijnen, Wim A M; Dullemont, Yolanda J; Schijven, Jack F; Hanzens-Brouwer, Anke J; Rosielle, Martine; Medema, Gertjan

    2007-05-01

    The decimal elimination capacity (DEC) of slow sand filtration (SSF) for Cryptosporidium parvum was assessed to enable quantitative microbial risk analysis of a drinking water production plant. A mature pilot plant filter of 2.56m(2) was loaded with C. parvum oocysts and two other persistent organisms as potential surrogates; spores of Clostridium perfringens (SCP) and the small-sized (4-7microm) centric diatom (SSCD) Stephanodiscus hantzschii. Highly persistent micro-organisms that are retained in slow sand filters are expected to accumulate and eventually break through the filter bed. To investigate this phenomenon, a dosing period of 100 days was applied with an extended filtrate monitoring period of 150 days using large-volume sampling. Based on the breakthrough curves the DEC of the filter bed for oocysts was high and calculated to be 4.7log. During the extended filtrate monitoring period the spatial distribution of the retained organisms in the filter bed was determined. These data showed little risk of accumulation of oocysts in mature filters most likely due to predation by zooplankton. The DEC for the two surrogates, SCP and SSCD, was 3.6 and 1.8log, respectively. On basis of differences in transport behaviour, but mainly because of the high persistence compared to the persistence of oocysts, it was concluded that both spores of sulphite-reducing clostridia (incl. SCP) and SSCD are unsuited for use as surrogates for oocyst removal by slow sand filters. Further research is necessary to elucidate the role of predation in Cryptosporidium removal and the fate of consumed oocysts. PMID:17400275

  11. Clostridium perfringens Alpha-Toxin Induces Gm1a Clustering and Trka Phosphorylation in the Host Cell Membrane

    PubMed Central

    Takagishi, Teruhisa; Oda, Masataka; Kabura, Michiko; Kurosawa, Mie; Tominaga, Kaori; Urano, Shiori; Ueda, Yoshibumi; Kobayashi, Keiko; Kobayashi, Toshihide; Sakurai, Jun; Terao, Yutaka; Nagahama, Masahiro

    2015-01-01

    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) with the yellow fluorescent protein-tagged C1AB domain of protein kinase C-? (EYFP-C1AB). DAG accumulated at the marginal region of the plasma membrane in alpha toxin-treated A549 cells, which also exhibited GM1a clustering and TrkA phosphorylation. Annexin V binding assays showed that alpha-toxin induced the exposure of phosphatidylserine on the outer leaflet of the plasma membrane. However, H148G, a variant toxin which binds cell membrane and has no enzymatic activity, did not induce DAG translocation, GM1a clustering, or TrkA phosphorylation. Alpha-toxin also specifically activated endogenous phospholipase C?-1 (PLC?-1), a TrkA adaptor protein, via phosphorylation. U73122, an endogenous PLC inhibitor, and siRNA for PLC?-1 inhibited the formation of DAG and release of IL-8. GM1a accumulation and TrkA phosphorylation in A549 cells treated with alpha-toxin were also inhibited by U73122. These results suggest that the flip-flop motion of hydrophobic lipids such as DAG leads to the accumulation of GM1a and TrkA. We conclude that the formation of DAG by alpha-toxin itself (first step) and activation of endogenous PLC?-1 (second step) leads to alterations in membrane dynamics, followed by strong phosphorylation of TrkA. PMID:25910247

  12. Use of Genetically Manipulated Strains of Clostridium perfringens Reveals that Both Alpha-Toxin and Theta-Toxin Are Required for Vascular Leukostasis To Occur in Experimental Gas Gangrene

    Microsoft Academic Search

    DARREN M. ELLEMOR; REBECCA N. BAIRD; MILENA M. AWAD; RICHARD L. BOYD; JULIAN I. ROOD; JOHN J. EMMINS

    1999-01-01

    A hallmark of gas gangrene (clostridial myonecrosis) pathology is a paucity of leukocytes infiltrating the necrotic tissue. The cause of this paucity most likely relates to the observation of leukocyte aggregates at the border of the area of tissue necrosis, often within the microvasculature itself. Infecting mice with genetically manipulated strains of Clostridium perfringens type A (deficient in either alpha-toxin

  13. The Clostridium perfringens Germinant Receptor Protein GerKC Is Located in the Spore Inner Membrane and Is Crucial for Spore Germination

    PubMed Central

    Banawas, Saeed; Paredes-Sabja, Daniel; Korza, George; Li, Yunfeng; Hao, Bing; Setlow, Peter

    2013-01-01

    The Gram-positive, anaerobic, spore-forming bacterium Clostridium perfringens causes a variety of diseases in both humans and animals, and spore germination is thought to be the first stage of C. perfringens infection. Previous studies have indicated that the germinant receptor (GR) proteins encoded by the bicistronic gerKA-gerKC operon as well as the proteins encoded by the gerKB and gerAA genes are required for normal germination of C. perfringens spores. We now report the individual role of these GR proteins by analyzing the germination of strains carrying mutations in gerKA, gerKC, or both gerKB and gerAA. Western blot analysis was also used to determine the location and numbers of GerKC proteins in spores. Conclusions from this work include the following: (i) gerKC mutant spores germinate extremely poorly with KCl, l-asparagine, a mixture of asparagine and KCl, or NaPi; (ii) gerKC spores germinate significantly more slowly than wild-type and other GR mutant spores with a 1:1 chelate of Ca2+ and dipicolinic acid and very slightly more slowly with dodecylamine; (iii) the germination defects in gerKC spores are largely restored by expressing the wild-type gerKA-gerKC operon in trans; (iv) GerKC is required for the spores' viability, almost certainly because of the gerKC spores' poor germination; and (v) GerKC is located in the spores' inner membrane, with ?250 molecules/spore. Collectively, these results indicate that GerKC is the main GR protein required for nutrient and nonnutrient germination of spores of C. perfringens food-poisoning isolates. PMID:24013629

  14. Toxin-associated and other genes in Clostridium perfringens type A isolates from bovine clostridial abomasitis (BCA) and jejunal hemorrhage syndrome (JHS).

    PubMed

    Schlegel, Benjamin J; Nowell, Victoria J; Parreira, Valeria R; Soltes, Glenn; Prescott, John F

    2012-10-01

    This study examined known or possible virulence-associated genes in type A Clostridium perfringens from cases of both bovine clostridial abomasitis (BCA) and jejunal hemorrhage syndrome (JHS) and compared these to isolates from calves that were healthy or had undifferentiated diarrheal illness. A real-time polymerase chain reaction (PCR) assay was used to genotype the 218 C. perfringens isolates. Isolates were sourced from healthy and diarrheic young and mature cattle (n = 191), from calves with confirmed or suspected BCA (n = 22), and from mature cattle with JHS (n = 5). Of 216 isolates (96%), 208 were positive for the cpa gene and 13% (29/218) were positive for atypical cpb2. Three of 8 (37.5%) confirmed BCA isolates, 2 of 13 (15.4%) suspected BCA isolates, and no JHS isolates tested positive for atypical cpb2. As all isolates were negative for cpb, cpb2, cpe, etx, netB, and tpeL, the results of the present study do not support a role for these genes in BCA or JHS. A subset of unique genes identified in 1 bovine clostridial abomasitis isolate (F262), for which a genome sequence is available, was searched for in 8 BCA isolates by PCR. None of the 10 genes was consistently present in all or even in a majority of BCA isolates. Many of these genes were also variably and inconsistently present in type A isolates from calves that did not have BCA. Although a virulence signature to aid in the diagnosis of BCA caused by C. perfringens type A was not identified, further work may discover a gene or group of genes that would constitute such a signature. PMID:23543949

  15. Toxin-associated and other genes in Clostridium perfringens type A isolates from bovine clostridial abomasitis (BCA) and jejunal hemorrhage syndrome (JHS)

    PubMed Central

    Schlegel, Benjamin J.; Nowell, Victoria J.; Parreira, Valeria R.; Soltes, Glenn; Prescott, John F.

    2012-01-01

    This study examined known or possible virulence-associated genes in type A Clostridium perfringens from cases of both bovine clostridial abomasitis (BCA) and jejunal hemorrhage syndrome (JHS) and compared these to isolates from calves that were healthy or had undifferentiated diarrheal illness. A real-time polymerase chain reaction (PCR) assay was used to genotype the 218 C. perfringens isolates. Isolates were sourced from healthy and diarrheic young and mature cattle (n = 191), from calves with confirmed or suspected BCA (n = 22), and from mature cattle with JHS (n = 5). Of 216 isolates (96%), 208 were positive for the cpa gene and 13% (29/218) were positive for atypical cpb2. Three of 8 (37.5%) confirmed BCA isolates, 2 of 13 (15.4%) suspected BCA isolates, and no JHS isolates tested positive for atypical cpb2. As all isolates were negative for cpb, cpb2, cpe, etx, netB, and tpeL, the results of the present study do not support a role for these genes in BCA or JHS. A subset of unique genes identified in 1 bovine clostridial abomasitis isolate (F262), for which a genome sequence is available, was searched for in 8 BCA isolates by PCR. None of the 10 genes was consistently present in all or even in a majority of BCA isolates. Many of these genes were also variably and inconsistently present in type A isolates from calves that did not have BCA. Although a virulence signature to aid in the diagnosis of BCA caused by C. perfringens type A was not identified, further work may discover a gene or group of genes that would constitute such a signature. PMID:23543949

  16. [Studies of necrotizing enteritis of suckling piglets (Clostridium perfringens type C enterotoxemia) in industrialized sow breeding units. 5. Control of the disease].

    PubMed

    Köhler, B; Zabke, J; Reiher, K; Rummler, H J

    1979-01-01

    Recent methods used and experience obtained in the control of necrotising enteritis are reported in this paper, with reference being made to both the pathogenesis and epizootiology of the disease. Two inoculations of the sows, using "Enterotoxämievakzine Dessau bivalent" five and three weeks before parturition, have worked well for prophylaxis. Oral treatment was applied to nursed piglets, using 40,000 I.U. of "Aviapen" and "V-Tablopen" penicillin per animal and day over periods between two and four days, helped to minimise piglet loss, particularly in the period between a fresh outbreak and full effectiveness of immunoprophylactic action. Such treatment was conducted metaphylactically and therapeutically. The first metaphylactic treatment was given within 24 hours from parturition. Combination of mother animal vaccination with the above therapeutic use of those two penicillin preparations worked extremely well in enzootically contaminated stocks and proved to be the most effective approach, for the time being, to controlling necrotising enteritis of nursed piglets. Yet, all those control measures failed to bring about full stock sanitation on industrialised units. Sow trading was not permitted until at least four weeks had elapsed from full effectiveness of mother animal vaccination, with the view to reducing the proliferation of Clostridium perfringens Type C via sales of breeding animals. All sows were given two "Enterotoxämievakzine Dessau bivalent" vaccinations, prior to sale. The animals were sold only to smaller farms (less than 500 sows for breeding) with concentional keeping patterns which were kept under constant diagnostic supervision. Neomycin, oxytetracycline, chloramphenicol, and other antibiotics against which Clostridium perfringens was resistant or in a position to assume resistance were used on endangered stocks only in conjunction with penicillin or not at all. This programme of control has proved to be efficient through a period of more than three years. PMID:232841

  17. Impact of Clean-Label Antimicrobials and Nitrite Derived from Natural Sources on the Outgrowth of Clostridium perfringens during Cooling of Deli-Style Turkey Breast.

    PubMed

    King, Amanda M; Glass, Kathleen A; Milkowski, Andrew L; Sindelar, Jeffrey J

    2015-05-01

    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 turkey breast to determine inhibition of Clostridium perfringens outgrowth during 15 h of chilling. Ten treatments of ground turkey breast (76% moisture, 1.2% salt) included a control and four antimicrobials: 1.0% tropical fruit extract, 0.7% dried vinegar, 1.0% cultured sugar-vinegar blend, and 2.0% lemon-vinegar blend. Each treatment was formulated without (uncured) and with nitrite (PCN; 50 ppm of NaNO2 from cultured celery juice powder). Treatments were inoculated with C. perfringens spores (three-strain mixture) to yield 2.5 log CFU/g. Individual 50-g portions were vacuum packaged, cooked to 71.1°C, and chilled from 54.4 to 26.7°C in 5 h and from 26.7 to 7.2°C in an additional 10 h. Triplicate samples were assayed for growth of C. perfringens at predetermined intervals by plating on tryptose-sulfite-cycloserine agar. Uncured control and PCN-only treatments allowed for 4.6- and 4.2-log increases at 15 h, respectively, and although all antimicrobial treatments allowed less outgrowth than uncured and PCN, the degree of inhibition varied. The 1.0% fruit extract and 1.0% cultured sugar-vinegar blend were effective at controlling populations at or below initial levels, whether or not PCN was included. Without PCN, 0.7% dried vinegar and 2.0% lemon-vinegar blend allowed for 2.0- and 2.5-log increases, respectively, and ?1.5-log increases with PCN. Results suggest using clean-label antimicrobials can provide for safe cooling following the study parameters, and greater inhibition of C. perfringens may exist when antimicrobials are used with nitrite. PMID:25951389

  18. Hazards associated with Clostridium perfringens in particular reference to predictive models applicable to cooling of cooked meat and poultry products

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The incidence of C. perfringens food-poisoning is quite common and costly. Although somewhat fastidious in growth characteristics using synthetic laboratory media, the microorganism is very prolific when found in food products. Inadequate cooling of foods in retail food operations is a major safety ...

  19. NanR, a Transcriptional Regulator That Binds to the Promoters of Genes Involved in Sialic Acid Metabolism in the Anaerobic Pathogen Clostridium perfringens

    PubMed Central

    Therit, Blair; Cheung, Jackie K.; Rood, Julian I.; Melville, Stephen B.

    2015-01-01

    Among many other virulence factors, Clostridium perfringens produces three sialidases NanH, NanI and NanJ. NanH lacks a secretion signal peptide and is predicted to be an intracellular enzyme, while NanI and NanJ are secreted. Previously, we had identified part of an operon encoding NanE (epimerase) and NanA (sialic acid lyase) enzymes. Further analysis of the entire operon suggests that it encodes a complete pathway for the transport and metabolism of sialic acid along with a putative transcriptional regulator, NanR. The addition of 30 mM N-acetyl neuraminic acid (Neu5Ac) to a semi-defined medium significantly enhanced the growth yield of strain 13, suggesting that Neu5Ac can be used as a nutrient. C. perfringens strain 13 lacks a nanH gene, but has NanI- and NanJ-encoding genes. Analysis of nanI, nanJ, and nanInanJ mutants constructed by homologous recombination revealed that the expression of the major sialidase, NanI, was induced by the addition of Neu5Ac to the medium, and that in separate experiments, the same was true of a nanI-gusA transcriptional fusion. For the nanI and nanJ genes, primer extension identified three and two putative transcription start sites, respectively. Gel mobility shift assays using purified NanR and DNA from the promoter regions of the nanI and nanE genes showed high affinity, specific binding by NanR. We propose that NanR is a global regulator of sialic acid-associated genes and that it responds, in a positive feedback loop, to the concentration of sialic acid in the cell. PMID:26197388

  20. LRP1 is a receptor for Clostridium perfringens TpeL toxin indicating a two-receptor model of clostridial glycosylating toxins.

    PubMed

    Schorch, Björn; Song, Shuo; van Diemen, Ferdy R; Bock, Hans H; May, Petra; Herz, Joachim; Brummelkamp, Thijn R; Papatheodorou, Panagiotis; Aktories, Klaus

    2014-04-29

    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 CROP domain but still intoxicates cells. Using a haploid genetic screen, we identified LDL receptor-related protein 1 (LRP1) as a host cell receptor for the TpeL toxin. LRP1-deficient cells are not able to take up TpeL and are not intoxicated. Expression of cluster IV of LRP1 is sufficient to rescue toxin uptake in these cells. By plasmon resonance spectroscopy, a KD value of 23 nM was determined for binding of TpeL to LRP1 cluster IV. The C terminus of TpeL (residues 1335-1779) represents the receptor-binding domain (RBD) of the toxin. RBD-like regions are conserved in all other clostridial glycosylating toxins preceding their CROP domain. CROP-deficient C. difficile toxin B is toxic to cells, depending on the RBD-like region (residues 1349-1811) but does not interact with LRP1. Our data indicate the presence of a second, CROP-independent receptor-binding domain in clostridial glycosylating toxins and suggest a two-receptor model for the cellular uptake of clostridial glycosylating toxins. PMID:24737893

  1. LRP1 is a receptor for Clostridium perfringens TpeL toxin indicating a two-receptor model of clostridial glycosylating toxins

    PubMed Central

    Schorch, Björn; Song, Shuo; van Diemen, Ferdy R.; Bock, Hans H.; May, Petra; Herz, Joachim; Brummelkamp, Thijn R.; Papatheodorou, Panagiotis; Aktories, Klaus

    2014-01-01

    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 CROP domain but still intoxicates cells. Using a haploid genetic screen, we identified LDL receptor-related protein 1 (LRP1) as a host cell receptor for the TpeL toxin. LRP1-deficient cells are not able to take up TpeL and are not intoxicated. Expression of cluster IV of LRP1 is sufficient to rescue toxin uptake in these cells. By plasmon resonance spectroscopy, a KD value of 23 nM was determined for binding of TpeL to LRP1 cluster IV. The C terminus of TpeL (residues 1335–1779) represents the receptor-binding domain (RBD) of the toxin. RBD-like regions are conserved in all other clostridial glycosylating toxins preceding their CROP domain. CROP-deficient C. difficile toxin B is toxic to cells, depending on the RBD-like region (residues 1349–1811) but does not interact with LRP1. Our data indicate the presence of a second, CROP-independent receptor-binding domain in clostridial glycosylating toxins and suggest a two-receptor model for the cellular uptake of clostridial glycosylating toxins. PMID:24737893

  2. The VirR Response Regulator from Clostridium perfringens Binds Independently to Two Imperfect Direct Repeats Located Upstream of the pfoA Promoter

    PubMed Central

    Cheung, Jackie K.; Rood, Julian I.

    2000-01-01

    Regulation of toxin production in the gram-positive anaerobe Clostridium perfringens occurs at the level of transcription and involves a two-component signal transduction system. The sensor histidine kinase is encoded by the virS gene, while its cognate response regulator is encoded by the virR gene. We have constructed a VirR expression plasmid in Escherichia coli and purified the resultant His-tagged VirR protein. Gel mobility shift assays demonstrated that VirR binds to the region upstream of the pfoA gene, which encodes perfringolysin O, but not to regions located upstream of the VirR-regulated plc, colA, and pfoR genes, which encode alpha-toxin, collagenase, and a putative pfoA regulator, respectively. The VirR binding site was shown by DNase I footprinting to be a 52-bp core sequence situated immediately upstream of the pfoA promoter. When this region was deleted, VirR was no longer able to bind to the pfoA promoter. The binding site was further localized to two imperfect direct repeats (CCCAGTTNTNCAC) by site-directed mutagenesis. Binding and protection analysis of these mutants indicated that VirR had the ability to bind independently to the two repeated sequences. Based on these observations it is postulated that the VirR positively regulates the synthesis of perfringolysin O by binding directly to a region located immediately upstream of the pfoA promoter and activating transcription. PMID:10613863

  3. Discrimination Efficacy of Fecal Pollution Detection in Different Aquatic Habitats of a High-Altitude Tropical Country, Using Presumptive Coliforms, Escherichia coli, and Clostridium perfringens Spores

    PubMed Central

    Byamukama, Denis; Mach, Robert L.; Kansiime, Frank; Manafi, Mohamad; Farnleitner, Andreas H.

    2005-01-01

    The performance of rapid and practicable techniques that presumptively identify total coliforms (TC), fecal coliforms (FC), Escherichia coli, and Clostridium perfringens spores (CP) by testing them on a pollution gradient in differing aquatic habitats in a high-altitude tropical country was evaluated during a 12-month period. Site selection was based on high and low anthropogenic influence criteria of paired sites including six spring, six stream, and four lakeshore sites spread over central and eastern parts of Uganda. Unlike the chemophysical water quality, which was water source type dependent (i.e., spring, lake, or stream), fecal indicators were associated with the anthropogenic influence status of the respective sites. A total of 79% of the total variability, including all the determined four bacteriological and five chemophysical parameters, could be assigned to either a pollution, a habitat, or a metabolic activity component by principal-component analysis. Bacteriological indicators revealed significant correlations to the pollution component, reflecting that anthropogenic contamination gradients were followed. Discrimination sensitivity analysis revealed high ability of E. coli to differentiate between high and low levels of anthropogenic influence. CP also showed a reasonable level of discrimination, although FC and TC were found to have worse discrimination efficacy. Nonpoint influence by soil erosion could not be detected during the study period by correlation analysis, although a theoretical contamination potential existed, as investigated soils in the immediate surroundings often contained relevant concentrations of fecal indicators. The outcome of this study indicates that rapid techniques for presumptive E. coli and CP determination may be reliable for fecal pollution monitoring in high-altitude tropical developing countries such as those of Eastern Africa. PMID:15640171

  4. Challenging the roles of CD44 and lipolysis stimulated lipoprotein receptor in conveying Clostridium perfringens iota toxin cytotoxicity in breast cancer

    PubMed Central

    2014-01-01

    Background 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 identify the mechanisms of iota toxin cytotoxicity in a tissue-specific, breast cancer model with the ultimate goal of laying the foundation for using iota toxin as a targeted breast cancer therapy. Methods In vitro model systems were used to determine the cytotoxic effect of iota toxin on breast cancer intrinsic subtypes. The use of overexpression and knockdown technologies confirmed the roles of LSR and CD44 in regulating iota toxin endocytosis and induction of cell death. Lastly, cytotoxicity assays were used to demonstrate the effect of iota toxin on a validated set of tamoxifen resistant breast cancer cell lines. Results Treatment of 14 breast cancer cell lines revealed that LSR+/CD44- lines were highly sensitive, LSR+/CD44+ lines were slightly sensitive, and LSR-/CD44+ lines were resistant to iota cytotoxicity. Reduction in LSR expression resulted in a significant decrease in toxin sensitivity; however, overexpression of CD44 conveyed toxin resistance. CD44 overexpression was correlated with decreased toxin-stimulated lysosome formation and decreased cytosolic levels of iota toxin. These findings indicated that expression of CD44 drives iota toxin resistance through inhibition of endocytosis in breast cancer cells, a role not previously defined for CD44. Moreover, tamoxifen-resistant breast cancer cells exhibited robust expression of LSR and were highly sensitive to iota-induced cytotoxicity. Conclusions Collectively, these data are the first to show that iota toxin has the potential to be an effective, targeted therapy for breast cancer. PMID:24990559

  5. Predictive model for growth of Clostridium perfringens during cooling of cooked beef supplemented with NaCl, sodium nitrite and sodium pyrophosphate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper presents a model for predicting relative growth of C. perfringens in ground beef products at different percentages of salt (0%, 1%, 2% and 3%) and nitrite (0 and 200 ppm). Included in the experiments were different levels of sodium pyrophosphate (SPP). The results of the experiments indic...

  6. A Possible Route for Foodborne Transmission of Clostridium difficile?

    PubMed Central

    Peck, Michael W.

    2015-01-01

    Abstract Spores of toxigenic Clostridium difficile and spores of food-poisoning strains of Clostridium perfringens show a similar prevalence in meats. Spores of both species are heat resistant and can survive cooking of foods. C. perfringens is a major cause of foodborne illness; studies are needed to determine whether C. difficile transmission by a similar route is a cause of infection. PMID:25599421

  7. Application of quantitative microbial risk assessments for estimation of risk management metrics: Clostridium perfringens in ready-to-eat and partially cooked meat and poultry products as an example.

    PubMed

    Crouch, Edmund A; Labarre, David; Golden, Neal J; Kause, Janell R; Dearfield, Kerry L

    2009-10-01

    The U.S. Department of Agriculture, Food Safety and Inspection Service is exploring quantitative risk assessment methodologies to incorporate the use of the Codex Alimentarius' newly adopted risk management metrics (e.g., food safety objectives and performance objectives). It is suggested that use of these metrics would more closely tie the results of quantitative microbial risk assessments (QMRAs) to public health outcomes. By estimating the food safety objective (the maximum frequency and/or concentration of a hazard in a food at the time of consumption) and the performance objective (the maximum frequency and/or concentration of a hazard in a food at a specified step in the food chain before the time of consumption), risk managers will have a better understanding of the appropriate level of protection (ALOP) from microbial hazards for public health protection. We here demonstrate a general methodology that allows identification of an ALOP and evaluation of corresponding metrics at appropriate points in the food chain. It requires a two-dimensional probabilistic risk assessment, the example used being the Monte Carlo QMRA for Clostridium perfringens in ready-to eat and partially cooked meat and poultry products, with minor modifications to evaluate and abstract required measures. For demonstration purposes, the QMRA model was applied specifically to hot dogs produced and consumed in the United States. Evaluation of the cumulative uncertainty distribution for illness rate allows a specification of an ALOP that, with defined confidence, corresponds to current industry practices. PMID:19833039

  8. Claudin-4 Overexpression in Epithelial Ovarian Cancer Is Associated with Hypomethylation and Is a Potential Target for Modulation of Tight Junction Barrier Function Using a C-Terminal Fragment of Clostridium perfringens Enterotoxin1

    PubMed Central

    Litkouhi, Babak; Kwong, Joseph; Lo, Chun-Min; Smedley, James G; McClane, Bruce A; Aponte, Margarita; Gao, Zhijian; Sarno, Jennifer L; Hinners, Jennifer; Welch, William R; Berkowitz, Ross S; Mok, Samuel C; Garner, Elizabeth I O

    2007-01-01

    Background Claudin-4, a tight junction (TJ) protein and receptor for the C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE), is overexpressed in epithelial ovarian cancer (EOC). Previous research suggests DNA methylation is a mechanism for claudin-4 overexpression in cancer and that C-CPE acts as an absorption-enhancing agent in claudin-4-expressing cells. We sought to correlate claudin-4 overexpression in EOC with clinical outcomes and TJ barrier function, investigate DNA methylation as a mechanism for overexpression, and evaluate the effect of C-CPE on the TJ. Methods Claudin-4 expression in EOC was quantified and correlated with clinical outcomes. Claudin-4 methylation status was determined, and claudin-4-negative cell lines were treated with a demethylating agent. Electric cell-substrate impedance sensing was used to calculate junctional (paracellular) resistance (Rb) in EOC cells after claudin-4 silencing and after C-CPE treatment. Results Claudin-4 overexpression in EOC does not correlate with survival or other clinical endpoints and is associated with hypomethylation. Claudin-4 overexpression correlates with Rb and C-CPE treatment of EOC cells significantly decreased Rb in a dose- and claudin-4-dependent noncytotoxic manner. Conclusions C-CPE treatment of EOC cells leads to altered TJ function. Further research is needed to determine the potential clinical applications of C-CPE in EOC drug delivery strategies. PMID:17460774

  9. Partial Characterization of an Enzyme Fraction with Protease Activity Which Converts the Spore Peptidoglycan Hydrolase (SleC) Precursor to an Active Enzyme during Germination of Clostridium perfringens S40 Spores and Analysis of a Gene Cluster Involved in the Activity

    PubMed Central

    Shimamoto, Seiko; Moriyama, Ryuichi; Sugimoto, Kazuhiro; Miyata, Shigeru; Makino, Shio

    2001-01-01

    A spore cortex-lytic enzyme of Clostridium perfringens S40 which is encoded by sleC is synthesized at an early stage of sporulation as a precursor consisting of four domains. After cleavage of an N-terminal presequence and a C-terminal prosequence during spore maturation, inactive proenzyme is converted to active enzyme by processing of an N-terminal prosequence with germination-specific protease (GSP) during germination. The present study was undertaken to characterize GSP. In the presence of 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS), a nondenaturing detergent which was needed for the stabilization of GSP, GSP activity was extracted from germinated spores. The enzyme fraction, which was purified to 668-fold by column chromatography, contained three protein components with molecular masses of 60, 57, and 52 kDa. The protease showed optimum activity at pH 5.8 to 8.5 in the presence of 0.1% CHAPS and retained activity after heat treatment at 55°C for 40 min. GSP specifically cleaved the peptide bond between Val-149 and Val-150 of SleC to generate mature enzyme. Inactivation of GSP by phenylmethylsulfonyl fluoride and HgCl2 indicated that the protease is a cysteine-dependent serine protease. Several pieces of evidence demonstrated that three protein components of the enzyme fraction are processed forms of products of cspA, cspB, and cspC, which are positioned in a tandem array just upstream of the 5? end of sleC. The amino acid sequences deduced from the nucleotide sequences of the csp genes showed significant similarity and showed a high degree of homology with those of the catalytic domain and the oxyanion binding region of subtilisin-like serine proteases. Immunochemical studies suggested that active GSP likely is localized with major cortex-lytic enzymes on the exterior of the cortex layer in the dormant spore, a location relevant to the pursuit of a cascade of cortex hydrolytic reactions. PMID:11371539

  10. Prevalence of cpb2, encoding beta2 toxin, in Clostridium perfringensfield isolates: correlation of genotype with phenotype

    Microsoft Academic Search

    Dawn M. Bueschel; B. Helen Jost; Stephen J. Billington; Hien T. Trinh; J. Glenn Songer

    2003-01-01

    Beta2 toxin, encoded by the cpb2 gene, has been implicated in the pathogenesis of porcine, equine and bovine enteritis by type A Clostridium perfringens. By incorporating primers to cpb2 into a multiplex genotyping PCR, we screened 3270 field isolates of C. perfringens. Of these, 37.2% were PCR positive for the cpb2 gene. The majority of isolates from cases of porcine

  11. epsilon. prime /. epsilon. : Review and recent progress

    SciTech Connect

    Franzini, P.J.

    1991-04-19

    The evolution of the theoretical perspective on {epsilon}{prime}/{epsilon} is reviewed. The introduction of the Z{sup O} penguin and the effects of high M{sub t} are discussed, in particular the possibility for {epsilon}{prime}/{epsilon} to be identically zero. Recent calculations of {epsilon}{prime}/{epsilon} based on current estimates and bounds on the input parameters are presented. 41 refs., 13 figs.

  12. Insights in metabolism and toxin production from the complete genome sequence of Clostridium tetani

    Microsoft Academic Search

    Holger Br; Gerhard Gottschalkb

    The decryption of prokaryotic genome sequences progresses rapidly and provides the scientific community with an enormous amount of information. Clostridial genome sequencing projects have been finished only recently, starting with the genome of the solvent-producing Clostridium acetobutylicum in 2001. A lot of attention has been devoted to the genomes of pathogenic clostridia. In 2002, the genome sequence of C. perfringens,

  13. Insights in metabolism and toxin production from the complete genome sequence of Clostridium tetani

    Microsoft Academic Search

    Holger Brüggemann; Gerhard Gottschalk

    2004-01-01

    The decryption of prokaryotic genome sequences progresses rapidly and provides the scientific community with an enormous amount of information. Clostridial genome sequencing projects have been finished only recently, starting with the genome of the solvent-producing Clostridium acetobutylicum in 2001. A lot of attention has been devoted to the genomes of pathogenic clostridia. In 2002, the genome sequence of C. perfringens,

  14. Antibacterial activity against Clostridium genus and antiradical activity of the essential oils from different origin.

    PubMed

    Ka?ániová, Miroslava; Vukovi?, Nenad; Horská, Elena; Salamon, Ivan; Bobková, Alica; Hleba, Lukáš; Fiskelová, Martina; Vat?ák, Alexander; Petrová, Jana; Bobko, Marek

    2014-01-01

    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 Clostridium ramosum. We obtained the original commercial essential oils samples of Lavandula angustifolia, Carum carvi, Pinus montana, Mentha piperita, Foeniculum vulgare Mill., Pinus sylvestris, Satureia montana, Origanum vulgare L. (2 samples), Pimpinella anisum, Rosmarinus officinalis L., Salvia officinalis L., Abies alba Mill., Chamomilla recutita L. Rausch and Thymus vulgaris L. produced in Slovakia (Calendula a.s., Nova Lubovna, Slovakia). The results of the disk diffusion method showed very high essential oils activity against all tested strains of microorganisms. The best antimicrobial activity against C. butyricum was found at Pimpinella anisum, against C. hystoliticum was found at Pinus sylvestris, against C. intestinale was found at Satureia hortensis L., against C. perfringens was found at Origanum vulgare L. and against C. ramosum was found at Pinus sylvestris. The results of broth microdilution assay showed that none of the essential oils was active against C. hystoliticum. The best antimicrobial activity against C. butyricum was found at Abies alba Mill., against C. intestinale was found at Abies alba Mill., against C. perfringens was found at Satureia montana and against C. ramosum was found at Abius alba and Carum carvi. Antioxidant DPPH radical scavenging activity was determined at several solutions of oil samples (50 ?L.mL(-1)-0.39 ?L.mL(-1)) and the best scavenging effect for the highest concentration (50 ?L.mL(-1)) was observed. The antioxidant properties were different in particular plant species. The highest% of inhibition after 30 min. of reaction was observed at Origanum vulgare (93%), Satureia montana (90.66%) and Lavandula augustifolia (90.22%). PMID:24813985

  15. Clostridium and Bacillus Binary Enterotoxins: Bad for the Bowels, and Eukaryotic Being

    PubMed Central

    Stiles, Bradley G.; Pradhan, Kisha; Fleming, Jodie M.; Samy, Ramar Perumal; Barth, Holger; Popoff, Michel R.

    2014-01-01

    Some pathogenic spore-forming bacilli employ a binary protein mechanism for intoxicating the intestinal tracts of insects, animals, and humans. These Gram-positive bacteria and their toxins include Clostridium botulinum (C2 toxin), Clostridium difficile (C. difficile toxin or CDT), Clostridium perfringens (?-toxin and binary enterotoxin, or BEC), Clostridium spiroforme (C. spiroforme toxin or CST), as well as Bacillus cereus (vegetative insecticidal protein or VIP). These gut-acting proteins form an AB complex composed of ADP-ribosyl transferase (A) and cell-binding (B) components that intoxicate cells via receptor-mediated endocytosis and endosomal trafficking. Once inside the cytosol, the A components inhibit normal cell functions by mono-ADP-ribosylation of globular actin, which induces cytoskeletal disarray and death. Important aspects of each bacterium and binary enterotoxin will be highlighted in this review, with particular focus upon the disease process involving the biochemistry and modes of action for each toxin. PMID:25198129

  16. Physiology and Sporulation in Clostridium.

    PubMed

    Dürre, Peter

    2014-08-01

    Clostridia are Gram-positive, anaerobic, endospore-forming bacteria, incapable of dissimilatory sulfate reduction. Comprising approximately 180 species, the genus Clostridium is one of the largest bacterial genera. Physiology is mostly devoted to acid production. Numerous pathways are known, such as the homoacetate fermentation by acetogens, the propionate fermentation by Clostridium propionicum, and the butyrate/butanol fermentation by C. acetobutylicum, a well-known solvent producer. Clostridia degrade sugars, alcohols, amino acids, purines, pyrimidines, and polymers such as starch and cellulose. Energy conservation can be performed by substrate-level phosphorylation as well as by the generation of ion gradients. Endospore formation resembles the mechanism elucidated in Bacillus. Morphology, contents, and properties of spores are very similar to bacilli endospores. Sporulating clostridia usually form swollen mother cells and accumulate the storage substance granulose. However, clostridial sporulation differs by not employing the so-called phosphorelay. Initiation starts by direct phosphorylation of the master regulator Spo0A. The cascade of sporulation-specific sigma factors is again identical to what is known from Bacillus. The onset of sporulation is coupled in some species to either solvent (acetone, butanol) or toxin (e.g., C. perfringens enterotoxin) formation. The germination of spores is often induced by various amino acids, often in combination with phosphate and sodium ions. In medical applications, C. butyricum spores are used as a C. difficile prophylaxis and as treatment against diarrhea. Recombinant spores are currently under investigation and testing as antitumor agents, because they germinate only in hypoxic tissues (i.e., tumor tissue), allowing precise targeting and direct killing of tumor cells. PMID:26104199

  17. epsilon'/epsilon in the Chiral Limit

    E-print Network

    Johan Bijnens; Joaquim Prades

    2000-05-18

    The $K\\to\\pi\\pi$ system is analyzed in the chiral limit within the Standard Model. We discuss how to connect the short-distance running in the $|\\Delta S|=1$ case to the matrix-elements calculated in a low-energy approximation in a scheme-independent fashion. We calculate this correction and the resulting Wilson Coefficients. The matrix elements are calculated to next-to-leading order in the $1/N_c$ expansion and combined with the Wilson coefficients to calculate the two isospin amplitudes and $\\epsilon_K^\\prime$. The $\\Delta I=1/2$ rule is reproduced within expected errors and we obtain a substantially larger value for $\\epsilon_K^\\prime/\\epsilon_K$ than most other analysises. We discuss the reasons for this difference. We also suggest that the $X$-boson method is an option for lattice QCD calculations.

  18. Small RNAs in the Genus Clostridium

    PubMed Central

    Chen, Yili; Indurthi, Dinesh C.; Jones, Shawn W.; Papoutsakis, Eleftherios T.

    2011-01-01

    The genus Clostridium includes major human pathogens and species important to cellulose degradation, the carbon cycle, and biotechnology. Small RNAs (sRNAs) are emerging as crucial regulatory molecules in all organisms, but they have not been investigated in clostridia. Research on sRNAs in clostridia is hindered by the absence of a systematic method to identify sRNA candidates, thus delegating clostridial sRNA research to a hit-and-miss process. Thus, we wanted to develop a method to identify potential sRNAs in the Clostridium genus to open up the field of sRNA research in clostridia. Using comparative genomics analyses combined with predictions of rho-independent terminators and promoters, we predicted sRNAs in 21 clostridial genomes: Clostridium acetobutylicum, C. beijerinckii, C. botulinum (eight strains), C. cellulolyticum, C. difficile, C. kluyveri (two strains), C. novyi, C. perfringens (three strains), C. phytofermentans, C. tetani, and C. thermocellum. Although more than one-third of predicted sRNAs have Shine-Dalgarno (SD) sequences, only one-sixth have a start codon downstream of SD sequences; thus, most of the predicted sRNAs are noncoding RNAs. Quantitative reverse transcription-PCR (Q-RT-PCR) and Northern analysis were employed to test the presence of a randomly chosen set of sRNAs in C. acetobutylicum and several C. botulinum strains, leading to the confirmation of a large fraction of the tested sRNAs. We identified a conserved, novel sRNA which, together with the downstream gene coding for an ATP-binding cassette (ABC) transporter gene, responds to the antibiotic clindamycin. The number of predicted sRNAs correlated with the physiological function of the species (high for pathogens, low for cellulolytic, and intermediate for solventogenic), but not with 16S rRNA-based phylogeny. PMID:21264064

  19. Latest experimental information on {epsilon}{prime}/{epsilon}

    SciTech Connect

    Yee B. Hsiung

    2000-08-17

    The authors review the latest experimental results in search for direct CP-violation by measuring the CP-violating parameters Re({epsilon}{prime}/{epsilon}) in neutral kaon decays. The recent result from Fermilab-KTeV Re({epsilon}{prime}/{epsilon}) = (28.0 {+-} 4.1) x 10{sup {minus}4}, and the new preliminary result from CERN-NA48 Re({epsilon}{prime}/{epsilon}) = (14.0 {+-} 4.3) x 10{sup {minus}4}, are presented. Both experiments, though using very different techniques, have now performed very well by collecting millions of events for all four relevant decay modes of K{sub L,S} to {pi}{sup +}{pi}{sup {minus}} and {pi}{sup 0}{pi}{sup 0} simultaneously. The current world average on this important measurement is Re({epsilon}{prime}/{epsilon}) = 19.3 {+-} 10{sup {minus}4} with a {chi}{sup 2}/ndf = 11.1/5, establishing the existence of direct CP-violation. The experimental status of such crucial measurements and the future prospects are also discussed here.

  20. Clostridium perfringens Bacteremia in an 85-Year-Old Diabetic Man

    PubMed Central

    Mirrakhimov, Aibek E.; Chandra, Gopika; Voore, Prakruthi; Khan, Maliha; Halytskyy, Oleksandr; Elhassan, Ahmed; Ali, Alaa M.

    2014-01-01

    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. PMID:25685130

  1. 9 CFR 113.112 - Clostridium Perfringens Type D Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...Purity test. Final container samples of completed product from each serial and each subserial shall be tested for viable bacteria and fungi as provided in § 113.26. (b) Safety test. Bulk or final container samples of completed product from...

  2. 9 CFR 113.111 - Clostridium Perfringens Type C Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...Purity test. Final container samples of completed product from each serial and each subserial shall be tested for viable bacteria and fungi as provided in § 113.26. (b) Safety test. Bulk or final container samples of completed product from...

  3. Enhancing Chicken Mucosal IgA Response Against Clostridium Perfringens a-toxin

    E-print Network

    Chen, Chang-Hsin 1977-

    2012-07-27

    antibodies in flow cytometry and immunohistochemistry (Gray et al., 2010). Unlike monoclonal antibodies, which are often produced in mammalian cell cultures, scFvs can be 21 in either prokaryotic or eukaryotic expression system (Peterson et al., 2006a...

  4. Electrotransformation of Clostridium thermocellum

    Microsoft Academic Search

    Michael V. Tyurin; Sunil G. Desai; Lee R. Lynd

    2004-01-01

    Electrotransformation of several strains of Clostridium thermocellum was achieved using plasmid pIKm1 with selection based on resistance to erythromycin and lincomycin. A custom-built pulse generator was used to apply a square 10-ms pulse to an electrotransformation cuvette consisting of a modified centrifuge tube. Transformation was verified by recovery of the shuttle plasmid pIKm1 from presumptive transformants of C. thermocellum with

  5. Diarylacylhydrazones: Clostridium-Selective Antibacterials with Activity Against Stationary-Phase Cells

    PubMed Central

    Casadei, Gabriele; Bremner, John B.; Lewis, Kim; Kelso, Michael J.

    2014-01-01

    Current antibiotics for treating Clostridium difficile infections (CDI), i.e. metronidazole, vancomycin and more recently fidaxomicin, are mostly effective but treatment failure and disease relapse remain as significant clinical problems. The shortcomings of these agents are attributed to their low selectivity for C. difficile over normal gut microflora and their ineffectiveness against C. difficile spores. This paper reports that certain diarylacylhydrazones identified during a high-throughput screening/counter-screening campaign show selective activity against two Clostridium species (C. difficile and C. perfringens) over common gut commensals. Representative examples are shown to possess activity similar to vancomycin against clinical C. difficile strains and to kill stationary-phase C. difficile cells, which are responsible for spore production. Structure-activity relationships with additional synthesised analogues suggested a protonophoric mechanism may play a role in the observed activity/selectivity and this was supported by the well-known protonophore carbonyl cyanide m-chlorophenyl hydrazone (CCCP) showing selective anti-Clostridium effects and activity similar to diarylacylhydrazones against stationary-phase C. difficile cells. Two diarylacylhydrazones were shown to be non-toxic towards human FaDu and Hep G2 cells indicating that further studies with the class are warranted towards new drugs for CDI. PMID:24360560

  6. Lactose-inducible system for metabolic engineering of Clostridium ljungdahlii.

    PubMed

    Banerjee, Areen; Leang, Ching; Ueki, Toshiyuki; Nevin, Kelly P; Lovley, Derek R

    2014-04-01

    The development of tools for genetic manipulation of Clostridium ljungdahlii has increased its attractiveness as a chassis for autotrophic production of organic commodities and biofuels from syngas and microbial electrosynthesis and established it as a model organism for the study of the basic physiology of acetogenesis. In an attempt to expand the genetic toolbox for C. ljungdahlii, the possibility of adapting a lactose-inducible system for gene expression, previously reported for Clostridium perfringens, was investigated. The plasmid pAH2, originally developed for C. perfringens with a gusA reporter gene, functioned as an effective lactose-inducible system in C. ljungdahlii. Lactose induction of C. ljungdahlii containing pB1, in which the gene for the aldehyde/alcohol dehydrogenase AdhE1 was downstream of the lactose-inducible promoter, increased expression of adhE1 30-fold over the wild-type level, increasing ethanol production 1.5-fold, with a corresponding decrease in acetate production. Lactose-inducible expression of adhE1 in a strain in which adhE1 and the adhE1 homolog adhE2 had been deleted from the chromosome restored ethanol production to levels comparable to those in the wild-type strain. Inducing expression of adhE2 similarly failed to restore ethanol production, suggesting that adhE1 is the homolog responsible for ethanol production. Lactose-inducible expression of the four heterologous genes necessary to convert acetyl coenzyme A (acetyl-CoA) to acetone diverted ca. 60% of carbon flow to acetone production during growth on fructose, and 25% of carbon flow went to acetone when carbon monoxide was the electron donor. These studies demonstrate that the lactose-inducible system described here will be useful for redirecting carbon and electron flow for the biosynthesis of products more valuable than acetate. Furthermore, this tool should aid in optimizing microbial electrosynthesis and for basic studies on the physiology of acetogenesis. PMID:24509933

  7. Lactose-Inducible System for Metabolic Engineering of Clostridium ljungdahlii

    PubMed Central

    Ueki, Toshiyuki; Nevin, Kelly P.; Lovley, Derek R.

    2014-01-01

    The development of tools for genetic manipulation of Clostridium ljungdahlii has increased its attractiveness as a chassis for autotrophic production of organic commodities and biofuels from syngas and microbial electrosynthesis and established it as a model organism for the study of the basic physiology of acetogenesis. In an attempt to expand the genetic toolbox for C. ljungdahlii, the possibility of adapting a lactose-inducible system for gene expression, previously reported for Clostridium perfringens, was investigated. The plasmid pAH2, originally developed for C. perfringens with a gusA reporter gene, functioned as an effective lactose-inducible system in C. ljungdahlii. Lactose induction of C. ljungdahlii containing pB1, in which the gene for the aldehyde/alcohol dehydrogenase AdhE1 was downstream of the lactose-inducible promoter, increased expression of adhE1 30-fold over the wild-type level, increasing ethanol production 1.5-fold, with a corresponding decrease in acetate production. Lactose-inducible expression of adhE1 in a strain in which adhE1 and the adhE1 homolog adhE2 had been deleted from the chromosome restored ethanol production to levels comparable to those in the wild-type strain. Inducing expression of adhE2 similarly failed to restore ethanol production, suggesting that adhE1 is the homolog responsible for ethanol production. Lactose-inducible expression of the four heterologous genes necessary to convert acetyl coenzyme A (acetyl-CoA) to acetone diverted ca. 60% of carbon flow to acetone production during growth on fructose, and 25% of carbon flow went to acetone when carbon monoxide was the electron donor. These studies demonstrate that the lactose-inducible system described here will be useful for redirecting carbon and electron flow for the biosynthesis of products more valuable than acetate. Furthermore, this tool should aid in optimizing microbial electrosynthesis and for basic studies on the physiology of acetogenesis. PMID:24509933

  8. Improved anatomy of epsilon'/epsilon in the Standard Model

    E-print Network

    Buras, Andrzej J; Jäger, Sebastian; Jamin, Matthias

    2015-01-01

    We present a new analysis of the ratio epsilon'/epsilon within the Standard Model (SM) using a formalism that is manifestly independent of the values of leading (V-A)x(V-A) QCD penguin, and EW penguin hadronic matrix elements of the operators Q_4, Q_9, and Q_10, and applies to the SM as well as extensions with the same operator structure. It is valid under the assumption that the SM exactly describes the data on CP-conserving K -> pi pi amplitudes. As a result of this and the high precision now available for CKM and quark mass parameters, to high accuracy epsilon'/epsilon depends only on two non-perturbative parameters, B_6^(1/2) and B_8^(3/2), and perturbatively calculable Wilson coefficients. Within the SM, we are separately able to determine the hadronic matrix element _0 from CP-conserving data, significantly more precisely than presently possible with lattice QCD. Employing B_6^(1/2) = 0.57+-0.15 and B_8^(3/2) = 0.76+-0.05, extracted from recent results by the RBC-UKQCD collaboration, we obtain epsilon'/...

  9. Phylogenomic analyses of clostridia and identification of novel protein signatures that are specific to the genus Clostridium sensu stricto (cluster I).

    PubMed

    Gupta, Radhey S; Gao, Beile

    2009-02-01

    The species of Clostridium comprise a very heterogeneous assemblage of bacteria that do not form a phylogenetically coherent group. It has been proposed previously that only a subset of the species of Clostridium that form a distinct cluster in the 16S rRNA tree (cluster I) should be regarded as the true representatives of the genus Clostridium (i.e. Clostridium sensu stricto). However, this cluster is presently defined only in phylogenetic terms, and no biochemical, molecular or phenotypic characteristic is known that is unique to species from this cluster. We report here phylogenomic and comparative analyses based on sequenced clostridial genomes in an attempt to bridge this gap and to clarify the evolutionary relationships among species of clostridia. In phylogenetic trees for species of clostridia based on concatenated sequences for 37 highly conserved proteins, the species of Clostridium cluster I formed a strongly supported clade that was separated from all other clostridia by a long branch. Several other Clostridium species that are not part of this cluster grouped reliably with other species of clostridia in a number of well-resolved clades. Our comparative genomic analyses have identified three conserved indels in three highly conserved proteins (a 4 aa insert in DNA gyrase A, a 1 aa deletion in ATP synthase beta subunit and a 1 aa insert in ribosomal protein S2) that are unique to the species of Clostridium cluster I and are not found in any other bacteria. blastp searches on various proteins in the genomes of Clostridium tetani E88 and Clostridium perfringens SM101 have also identified more than 10 proteins that are found uniquely in the cluster I species. These results provide evidence that the species of Clostridium cluster I not only are phylogenetically distinct but also share many unique molecular characteristics. These newly identified molecular markers provide useful tools to define and circumscribe the genus Clostridium sensu stricto in more definitive terms. We have also identified a 7-9 aa conserved insert in the enzyme phosphoglycerate dehydrogenase that is uniquely found in the Clostridium thermocellum, Thermoanaerobacter pseudethanolicus, Thermoanaerobacter tengcogensis and Caldicellulosiruptor saccharolyticus homologues, and is absent from all other bacteria. These species form a well-defined clade in the phylogenetic trees and this indel provides a potential molecular marker for this clostridial cluster. PMID:19196767

  10. Improved anatomy of epsilon'/epsilon in the Standard Model

    E-print Network

    Andrzej J. Buras; Martin Gorbahn; Sebastian Jäger; Matthias Jamin

    2015-07-22

    We present a new analysis of the ratio epsilon'/epsilon within the Standard Model (SM) using a formalism that is manifestly independent of the values of leading (V-A)x(V-A) QCD penguin, and EW penguin hadronic matrix elements of the operators Q_4, Q_9, and Q_10, and applies to the SM as well as extensions with the same operator structure. It is valid under the assumption that the SM exactly describes the data on CP-conserving K -> pi pi amplitudes. As a result of this and the high precision now available for CKM and quark mass parameters, to high accuracy epsilon'/epsilon depends only on two non-perturbative parameters, B_6^(1/2) and B_8^(3/2), and perturbatively calculable Wilson coefficients. Within the SM, we are separately able to determine the hadronic matrix element _0 from CP-conserving data, significantly more precisely than presently possible with lattice QCD. Employing B_6^(1/2) = 0.57+-0.15 and B_8^(3/2) = 0.76+-0.05, extracted from recent results by the RBC-UKQCD collaboration, we obtain epsilon'/epsilon = (2.2+-3.7) 10^-4, substantially more precise than the recent RBC-UKQCD prediction and more than 3 sigma below the experimental value (16.6+-2.3) 10^-4, with the error being fully dominated by that on B_6^(1/2). Even discarding lattice input completely, but employing the recently obtained bound B_6^(1/2) anatomy of the various SM uncertainties, including all sub-leading hadronic matrix elements, briefly commenting on the possibility of underestimated SM contributions as well as on the impact of our results on new physics models.

  11. CDP-diacylglycerol synthase activity in Clostridium perfingens

    SciTech Connect

    Carmen, G.M.; Zaniewski, R.L.; Cousminer, J.J.

    1982-01-01

    CTP: phosphatidate cytidylyltransferase (CDP-diacylglycerol synthase; EC 2.7.7.41) was identified in the cell envelope fraction of the gram-positive anaerobe Clostridium perfringens. The association of this enzyme with the cell envelope fraction of cell extracts was demonstrated by glycerol density gradient centrifugation and by activity sedimenting with the 100,000 x g pellet. The enzyme exhibited a broad pH optimium between pH 6.5 and pH 7.5. Enzyme activity was dependent on magnesium (5 mM) or manganese (1 mM) ions. Activity was also dependent on the addition on the nonionic detergent Triton X-100 (5 mM). The apparent Km values for CTP and phosphatidic acid were 0.18 mM and 0.22 mM respectively. Thioreactive agents inhibited activity, indicating that a sulfhydryl group is essential for activity. Maximal enzyme activity was observed at 50 degrees C. (Refs. 24).

  12. Facteurs Epsilon p-adiques Adriano Marmora

    E-print Network

    Franchi, Jacques

    Facteurs Epsilon p-adiques Adriano Marmora Abstract We develop and study the epsilon factor of a local system of p-adic coecients over the spectrum of a complete discrete valuation eld K with nite residue eld of characteristic p > 0. In the equal characteristic case, we dene the epsilon factor

  13. Genomics of Clostridium tetani.

    PubMed

    Brüggemann, Holger; Brzuszkiewicz, Elzbieta; Chapeton-Montes, Diana; Plourde, Lucile; Speck, Denis; Popoff, Michel R

    2015-05-01

    Genomic information about Clostridium tetani, the causative agent of the tetanus disease, is scarce. The genome of strain E88, a strain used in vaccine production, was sequenced about 10 years ago. One additional genome (strain 12124569) has recently been released. Here we report three new genomes of C. tetani and describe major differences among all five C. tetani genomes. They all harbor tetanus-toxin-encoding plasmids that contain highly conserved genes for TeNT (tetanus toxin), TetR (transcriptional regulator of TeNT) and ColT (collagenase), but substantially differ in other plasmid regions. The chromosomes share a large core genome that contains about 85% of all genes of a given chromosome. The non-core chromosome comprises mainly prophage-like genomic regions and genes encoding environmental interaction and defense functions (e.g. surface proteins, restriction-modification systems, toxin-antitoxin systems, CRISPR/Cas systems) and other fitness functions (e.g. transport systems, metabolic activities). This new genome information will help to assess the level of genome plasticity of the species C. tetani and provide the basis for detailed comparative studies. PMID:25638019

  14. Distribution of Clostridium botulinum.

    PubMed Central

    Huss, H H

    1980-01-01

    The distribution of Clostridium botulinum in the natural environments of Denmark, The Faroe Islands, Iceland, Greenland, and Bangladesh was examined. A total of 684 samples were tested. Type E was found in 90% of samples from the aquatic environment of Denmark, including sediments from young artificial lakes, and in 86% of samples from the marine environment of Greenland. Type E was not found in Danish cultivated soil and woodlands, including cultivated soil from reclaimed sea beds, but type B was frequently demonstrated in these environments. C. botulinum types A, B, or E were found in 2.6% of samples from the environments of the Faroe Islands and Iceland, whereas types C or D were demonstrated in 42% of samples from Bangladesh. The incidence of type E in aquatic sediments was not related to general industrial pollution or a high content of rotting vegetation. Fish or a rich aquatic fauna, on the other hand, appeared to contribute to a high incidence of type E. Based on these findings, it is suggested that type E is a true aquatic organism, because this environment offers the best conditions for survival of the spore in nature. It is further suggested that its presence in aquatic bottom deposits is based on sedimentation after proliferation in the carrion of the aquatic fauna and dissemination by water currents and migrating fish. PMID:6990867

  15. Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp.

    PubMed

    Ahn, Y J; Park, S J; Lee, S G; Shin, S C; Choi, D H

    2000-07-01

    The growth responses of nine human intestinal bacteria to liquid culture of Cordyceps militaris Link. Pt. (Ascomycotina: Clavicipitaceae) collected from a pupa of Bombyx mori L. (Lepidoptera: Bombycidae) were examined using spectrophotometric and impregnated paper disk methods and compared to those of tetracycline and chloramphenicol, as well as those of Coptis japonica root-derived berberine chloride. The biologically active constituent of the cultures was characterized as cordycepin (3'-deoxyadenosine) by spectroscopic analysis. This compound revealed potent growth-inhibiting activity toward Clostridium paraputrificum and Clostridium perfringens at 10 microgram/disk without adverse effects on the growth of Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Lactobacillus acidophilus, and Lactobacillus casei, whereas tetracycline and chloramphenicol inhibited the growth of these lactic acid-producing bacteria, clostridia and Escherichia coli. However, C. militaris-derived materials revealed no growth stimulation on the bifidobacteria and lactobacilli. These results may be an indication of at least one of the pharmacological actions of C. militaris. As a naturally occurring antibacterial agent, cordycepin could be useful as a new preventive agent against various diseases caused by clostridia. PMID:10898616

  16. epsilon'/epsilon and Rare K and B Decays in the MSSM

    E-print Network

    A. J. Buras; P. Gambino; M. Gorbahn; S. Jager; L. Silvestrini

    2000-07-27

    We analyze the CP violating ratio epsilon'/epsilon and rare K and B decays in the MSSM with minimal flavour and CP violation, including NLO QCD corrections and imposing constraints on the supersymmetric parameters coming from epsilon, B_{d,s}^0-\\bar B_{d,s}^0 mixings, B to X_s gamma, Delta rho in the electroweak precision studies and from the lower bound on the neutral Higgs mass. We provide a compendium of phenomenologically relevant formulae in the MSSM. Denoting by T(Q) the MSSM prediction for a given quantity normalized to the Standard Model result we find the ranges: 0.53 epsilon'/epsilon) K^+ to pi^+ nu nubar) K_L to pi^0 nu nubar) K_L to pi^0 e^+ e^-) epsilon'/epsilon in the SM and find in NDR epsilon'/epsilon = (9.2^{+6.8}_{-4.0}), a value 15 % higher than in our 1999 analysis.

  17. Interspecies Conversion of Clostridium botulinum Type C to Clostridium novyi Type A by Bacteriophage

    Microsoft Academic Search

    M. W. Eklund; F. T. Poysky; J. A. Meyers; G. A. Pelroy

    1974-01-01

    When Clostridium botulinum type C is cured of its prophage it simultaneously ceases to produce toxin. This nontoxigenic culture can then be converted to another toxigenic bacterial species, Clostridium novyi type A or to toxigenic Clostridium botulinum types C or D, by specific bacteriophages. The toxigenicity and type of toxin produced by these cultures depends upon the continued presence of

  18. Small Weak Epsilon-Nets Boris Aronov

    E-print Network

    Aurenhammer, Franz

    Small Weak Epsilon-Nets Boris Aronov Franz Aurenhammer Ferran Hurtado Stefan Langerman§ David , the largest epsilon that can be guaranteed for any P when |Q| = i, for small values of i. 1 Introduction Let P Bruxelles, Belgium, stefan.langerman@ulb.ac.be. ¶ School of Computing, Queen's University, Kingston, ON K7L

  19. Genetic Manipulation of Clostridium difficile

    PubMed Central

    Bouillaut, Laurent; McBride, Shonna M.; Sorg, Joseph A.

    2012-01-01

    Clostridium difficile is a Gram-positive, spore forming, anaerobic, intestinal bacterium and is the most common cause of antibiotic-associated colitis. For many years this organism was considered genetically intractable, but in the past 10 years, multiple methods have been developed or adapted for genetic manipulation of C. difficile. This unit describes the molecular techniques used for genetic modification of this organism, including methods for gene disruption, complementation, plasmid introduction and integration, and cross-species conjugations. PMID:21400677

  20. Production by Clostridium spiroforme of an iotalike toxin that possesses mono(ADP-ribosyl)transferase activity: identification of a novel class of ADP-ribosyltransferases.

    PubMed Central

    Simpson, L L; Stiles, B G; Zepeda, H; Wilkins, T D

    1989-01-01

    Clostridium spiroforme iotalike toxin produced time- and concentration-dependent incorporation of ADP-ribose into homo-poly-L-arginine. Polyasparagine, polyglutamic acid, polylysine, and agmatine were poor substrates. Enzyme activity was associated with the light-chain polypeptide of the toxin. The heavy chain did not possess ADP-ribosyltransferase activity, nor did it enhance or inhibit activity of the light chain. In broken-cell assays, the toxin acted mainly on G-actin, rather than F-actin. A single ADP-ribose group was transferred to each substrate molecule (G-actin). The enzyme was heat sensitive, had a pH optimum in the range of 7 to 8, was inhibited by high concentrations of nicotinamide, and was reversibly denatured by urea and guanidine. Physiological levels of nucleotides (AMP, ADP, ATP, and ADP-ribose) and cations (Na+, K+, Ca2+, and Mg2+) were not very active as enzyme inhibitors. The toxin was structurally and functionally similar to Clostridium botulinum type C2 toxin and Clostridium perfringens iota toxin. When combined with previous findings, the data suggest that a new class of mono(ADP-ribosyl)ating toxins has been found and that these agents belong to a related and possibly homologous series of binary toxins. Images PMID:2521214

  1. Introduction to the improved functional epsilon algorithm

    NASA Astrophysics Data System (ADS)

    Thukral, R.

    2002-10-01

    This paper introduces the improved functional epsilon algorithm. We have defined this new method in principle of the modified Aitken [Delta]2 algorithm. Moreover, we have found that the improved functional epsilon algorithm has remarkable precision of the approximation of the exact solution and there exists a relationship with the integral Padé approximant. The use of the improved functional epsilon algorithm for accelerating the convergence of sequence of functions is demonstrated. The relationship of the improved functional epsilon algorithm with the integral Padé approximant is also demonstrated. Moreover, we illustrate the similarity between the integral Padé approximant and the modified Aitken [Delta]2 algorithm; thus we have shown that the integral Padé approximant is a natural generalisation of modified Aitken [Delta]2 algorithm.

  2. Cloning and nucleotide sequence analysis of the colH gene from Clostridium histolyticum encoding a collagenase and a gelatinase.

    PubMed Central

    Yoshihara, K; Matsushita, O; Minami, J; Okabe, A

    1994-01-01

    The colH gene encoding a collagenase was cloned from Clostridium histolyticum JCM 1403. Nucleotide sequencing showed a major open reading frame encoding a 116-kDa protein of 1,021 amino acid residues. The deduced amino acid sequence contains a putative signal sequence and a zinc metalloprotease consensus sequence, HEXXH. A 116-kDa collagenase and a 98-kDa gelatinase were copurified from culture supernatants of C. histolyticum. While the former degraded both native and denatured collagen, the latter degraded only denatured collagen. Peptide mapping with V8 protease showed that all peptide fragments, except a few minor ones, liberated from the two enzymes coincided with each other. Analysis of the N-terminal amino acid sequence of the two enzymes revealed that their first 24 amino acid residues were identical and coincided with those deduced from the nucleotide sequence. These results indicate that the 98-kDa gelatinase is generated from the 116-kDa collagenase by cleaving off the C-terminal region, which could be responsible for binding or increasing the accessibility of the collagenase to native collagen fibers. The role of the C-terminal region in the functional and evolutional aspects of the collagenase was further studied by comparing the amino acid sequence of the C. histolyticum collagenase with those of three homologous enzymes: the collagenases from Clostridium perfringens and Vibrio alginolyticus and Achromobacter lyticus protease I. Images PMID:7961400

  3. epsilon'/epsilon and Rare K Decays in the Standard Model and Supersymmetry

    E-print Network

    L. Silvestrini

    2000-09-25

    After briefly reviewing the status of epsilon'/epsilon in the Standard Model, I discuss SUSY contributions to epsilon'/epsilon, K^+ to pi^+ nu bar nu, K_L to pi^0 nu bar nu and K_L to pi^0 e^+ e^-. While in the simplest case of the MSSM with Minimal Flavour Violation the main effect is a suppression of these transitions with respect to the Standard Model, large enhancements are possible in more general SUSY models, with interesting correlations among the different processes.

  4. Pi Mu Epsilon New York Alpha Chapter Membership Application

    E-print Network

    McConnell, Terry

    Pi Mu Epsilon New York Alpha Chapter Membership Application (Due by March 28, 2014) Contact dghsu opinion, should Pi Mu Epsilon focus on next year as an organization? 2) In what ways can Pi Mu Epsilon for admission to Pi Mu Epsilon in the coming years? 4) What motivates you to participate in the study

  5. Binary Bacterial Toxins: Biochemistry, Biology, and Applications of Common Clostridium and Bacillus Proteins

    PubMed Central

    Barth, Holger; Aktories, Klaus; Popoff, Michel R.; Stiles, Bradley G.

    2004-01-01

    Certain pathogenic species of Bacillus and Clostridium have developed unique methods for intoxicating cells that employ the classic enzymatic “A-B” paradigm for protein toxins. The binary toxins produced by B. anthracis, B. cereus, C. botulinum, C. difficile, C. perfringens, and C. spiroforme consist of components not physically associated in solution that are linked to various diseases in humans, animals, or insects. The “B” components are synthesized as precursors that are subsequently activated by serine-type proteases on the targeted cell surface and/or in solution. Following release of a 20-kDa N-terminal peptide, the activated “B” components form homoheptameric rings that subsequently dock with an “A” component(s) on the cell surface. By following an acidified endosomal route and translocation into the cytosol, “A” molecules disable a cell (and host organism) via disruption of the actin cytoskeleton, increasing intracellular levels of cyclic AMP, or inactivation of signaling pathways linked to mitogen-activated protein kinase kinases. Recently, B. anthracis has gleaned much notoriety as a biowarfare/bioterrorism agent, and of primary interest has been the edema and lethal toxins, their role in anthrax, as well as the development of efficacious vaccines and therapeutics targeting these virulence factors and ultimately B. anthracis. This review comprehensively surveys the literature and discusses the similarities, as well as distinct differences, between each Clostridium and Bacillus binary toxin in terms of their biochemistry, biology, genetics, structure, and applications in science and medicine. The information may foster future studies that aid novel vaccine and drug development, as well as a better understanding of a conserved intoxication process utilized by various gram-positive, spore-forming bacteria. PMID:15353562

  6. Molecular dynamics simulations for pure epsilon-CL-20 and epsilon-CL-20-based PBXs.

    PubMed

    Xu, Xiao-Juan; Xiao, He-Ming; Xiao, Ji-Jun; Zhu, Wei; Huang, Hui; Li, Jin-Shan

    2006-04-13

    Molecular dynamics has been employed to simulate the well-known high energy density compound epsilon-CL-20 (hexanitrohexaazaisowurtzitane) crystal and 12 epsilon-CL-20-based PBXs (polymer bonded explosives) with four kinds of typical fluorine polymers, i.e., polyvinylidenedifluoride, polychlorotrifluoroethylene, fluorine rubber (F(2311)), and fluorine resin (F(2314)) individually. The elastic coefficients, isotropic mechanical properties (tensile moduli, bulk moduli, shear moduli, and poission's ratios), and bonding energy are first reported for epsilon-CL-20 crystal and epsilon-CL-20-based polymer bonded explosives (PBXs). The mechanical properties of epsilon-CL-20 can be effectively improved by blending with a small amount of fluorine polymers, and the whole effect of the adding fluorine polymers to improve mechanical properties of PBXs along the three crystalline surfaces of epsilon-CL-20 is found to be (100) approximately (001) > (010). The interaction between each of the crystalline surfaces and each of the fluorine polymers is different, and the ordering of binding energy for the three surfaces is (001) > (100) > (010); F(2314) always has the strongest binding ability with the three different surfaces. F(2314) can best improve the ductibility and tenacity of PBX when it is positioned on epsilon-CL-20 (001) crystal surface. The calculations on detonation performances for pure epsilon-CL-20 crystal and the four epsilon-CL-20-based PBXs show that adding a small amount of fluorine polymer into pure epsilon-CL-20 will lower detonation performance, but each detonation parameter of the obtained PBXs is still excellent. PMID:16599487

  7. {epsilon}{prime}/{epsilon} and rare KL decays from KTeV Experiment

    SciTech Connect

    Yee Bob Hsiung

    2001-01-16

    The authors update the current status for the measurement of the direct-CP violation parameters {epsilon}{prime}/{epsilon} in the KTeV experiment at Fermilab. Substantial statistics have been accumulated during the 1996-7 run and 1999 run for both {epsilon}{prime}/{epsilon} and rare K{sub L} decay searches. The first KTeV result on {epsilon}{prime}/{epsilon} published last year was Re({epsilon}{prime}/{epsilon}) = (28.0 {+-} 4.1) x 10{sup {minus}4} based on the 23% data from 1996-7 run. Combining with the previous E731, NA31 and the recent preliminary NA48 results, the grand average is Re({epsilon}{prime}/{epsilon}) = (19.3 {+-} 3.6) x 10{sup {minus}4} (with S = 1:49), more than 5{sigma} above zero. More data from both KTeV and NA48 after completing the analysis will further improve the precision of this measurement in the near future and hopefully further improve the agreement. New results on the branching ratio and form factor measurements of K{sub L} {r_arrow} {mu}{sup +} {mu}{sup {minus}} {gamma} using 1997 data are also presented. We find that BR(K{sub L} {r_arrow} {mu}{sup +} {mu}{sup {minus}} {gamma}) = (3.66 {+-} 0.04{sub stat} {+-} 0.07{sub syst}) x 10{sup {minus}7}. The form factor parameter {alpha}{sub K}* is measured to be {alpha}{sub K}* = {minus}0.157{sub {minus}0.027}{sup +0.025}. We make the first measurement of the parameter {alpha} from the D'Ambrosio, Isidori, and Portoles form factor, finding {alpha} = {minus}1.53 {+-} 0.09. This measurement of {alpha} limits the CKM parameter {rho} > {minus}0.2.

  8. Characterization of six murine monoclonal antibodies specific for toxin B of Clostridium difficile.

    PubMed

    Coughlin, R T; Annunziato, M; Roberson, J; Marciani, D J

    1994-04-01

    Six murine hybridoma cell lines producing monoclonal antibodies (MAbs) specific for Toxin B of Clostridium difficile have been generated from toxin-immunized female RBF/DnJ mice. All six antibodies were reactive in Western blots with a > 200-kD protein in the supernatants of the toxigenic strain 10463 and were unreactive with similarly prepared material from the nontoxigenic strain 2037. Polyclonal antisera from rabbits immunized with Toxin B reacted on Western blots primarily with Toxin B, a 40-kD and a 55-kD band with a minor set of triplet bands at approximately 100 kD. None of the MAbs reacted in a direct EIA with purified Toxin A from C. difficile but two MAbs reacted weakly with a trypsin-sensitive band (> 200 kD) in Western blots of C. sordellii. Polyclonal antisera developed against Toxin B reacted strongly with supernatants from C. sordellii, C. bifermentans, and the nontoxigenic strain 2037. Toxin B-specific antisera was unreactive with supernatants from C. perfringens or purified Toxin A from C. difficile in direct EIA. Toxin B-specific MAbs linked to an affinity column were able to deplete bacterial supernatant of cytotoxigenic activity. PMID:8050780

  9. SleC Is Essential for Germination of Clostridium difficile Spores in Nutrient-Rich Medium Supplemented with the Bile Salt Taurocholate ?

    PubMed Central

    Burns, David A.; Heap, John T.; Minton, Nigel P.

    2010-01-01

    Clostridium difficile is the major cause of infectious diarrhea and a major burden to health care services. The ability of this organism to form endospores plays a pivotal role in infection and disease transmission. Spores are highly resistant to many forms of disinfection and thus are able to persist on hospital surfaces and disseminate infection. In order to cause disease, the spores must germinate and the organism must grow vegetatively. Spore germination in Bacillus is well understood, and genes important for this process have recently been identified in Clostridium perfringens; however, little is known about C. difficile. Apparent homologues of the spore cortex lytic enzyme genes cwlJ and sleB (Bacillus subtilis) and sleC (C. perfringens) are present in the C. difficile genome, and we describe inactivation of these homologues in C. difficile 630?erm and a B1/NAP1/027 clinical isolate. Spores of a sleC mutant were unable to form colonies when germination was induced with taurocholate, although decoated sleC spores formed the same number of heat-resistant colonies as the parental control, even in the absence of germinants. This suggests that sleC is absolutely required for conversion of spores to vegetative cells, in contrast to CD3563 (a cwlJ/sleB homologue), inactivation of which had no effect on germination and outgrowth of C. difficile spores under the same conditions. The B1/NAP1/027 strain R20291 was found to sporulate more slowly and produce fewer spores than 630?erm. Furthermore, fewer R20291 spores germinated, indicating that there are differences in both sporulation and germination between these epidemic and nonepidemic C. difficile isolates. PMID:19933358

  10. Clostridium difficile in paediatric populations

    PubMed Central

    Allen, Upton D

    2014-01-01

    An increase in Clostridium difficile infection incidence has been observed among hospitalized children in the United States. The present statement, targeted at clinicians caring for infants and children in community and institutional settings, summarizes the relevant information relating to the role of C difficile in childhood diarrhea and provides recommendations for diagnosis, prevention and treatment. Significant differences between adult and paediatric risk factors and disease are discussed, along with emerging therapies. The relationship between age and disease severity in children with a newly emergent and more fluoroqinolone-resistant strain of C difficile (North American Pulse-field type-1 [NAP1]) remains unknown. The importance of antimicrobial stewardship as a preventive strategy is highlighted. This statement replaces a previous Canadian Paediatric Society position statement on C difficile published in 2000. PMID:24627655

  11. Biofilm formation by Clostridium difficile

    PubMed Central

    Dapa, Tanja; Unnikrishnan, Meera

    2013-01-01

    Clostridium difficile infection (CDI) is a major healthcare-associated disease worldwide. Recurring infections and increasing antibiotic resistance have complicated treatment of CDI. While C. difficile spores are important for transmission and persistence of CDI, other factors such as gut colonization and formation of bacterial communities in the gut may also contribute to pathogenesis and persistence, but have not been well investigated. Recently, we reported that important clinical C. difficile strains are able to form composite biofilms in vitro. C. difficile biofilm formation is a complex process, modulated by several different factors, including cell surface components and regulators. We also reported that bacteria within biofilms are more resistant to high concentrations of vancomycin, the antibiotic of choice for treatment of CDI. Here we summarize our recent findings and discuss the implications of biofilm formation by this anaerobic gut pathogen in disease pathogenesis and treatment. PMID:23892245

  12. The Final Measurement of Epsilon'/Epsilon from KTeV

    E-print Network

    E. T. Worcester

    2009-09-14

    We present precise measurements of CP and CPT symmetry based on the full dataset of K to 2pi decays collected by the KTeV experiment at FNAL. We measure the direct CP violation parameter Re(epsilon'/epsilon) = (19.2 pm 2.1)x10-4. We also report the KL-KS mass difference, the KS lifetime, the phase of epsilon, and the phase difference (phi00)-(phi+-). These results are consistent with other experimental results and with CPT symmetry.

  13. The Final Measurement of Epsilon'/Epsilon from KTeV

    E-print Network

    E. T. Worcester

    2009-10-16

    We present precise measurements of CP and CPT symmetry based on the full dataset of K to 2pi decays collected by the KTeV experiment at FNAL. We measure the direct CP violation parameter Re(epsilon'/epsilon) = (19.2 pm 2.1)x10-4. We also report the KL-KS mass difference, the KS lifetime, the phase of epsilon, and the phase difference (phi00)-(phi+-). These results are consistent with other experimental results and with CPT symmetry.

  14. Clostridium difficile infection in patients with inflammatory bowel disease

    PubMed Central

    Biesiada, Gra?yna; Perucki, William; Mach, Tomasz

    2014-01-01

    Clostridium difficile is a bacterium widely distributed in the human environment. In the last decade the incidence and severity of Clostridium difficile infection has grown, particularly in Europe and North America, making it one of the more common nosocomial infections. A group particularly susceptible to Clostridium difficile infection are patients with inflammatory bowel disease, especially those with involvement of the colon. This paper presents relevant data on Clostridium difficile infections in inflammatory bowel disease patients, including epidemiology, pathogenesis, diagnosis and treatment. PMID:25097707

  15. Epsilon Stereo Pairs Yuanyuan Ding Jingyi Yu

    E-print Network

    Taufer, Michela

    University of Delaware Newark, DE 19716, USA {ding,yu}@cis.udel.edu Abstract Human stereo vision worksEpsilon Stereo Pairs Yuanyuan Ding Jingyi Yu Department of Computer and Information Sciences that very few vari- eties of multiperspective stereo pairs exist. In this paper, we introduce a new stereo

  16. Charter Members History of Pi Mu Epsilon

    E-print Network

    Feingold, Alex

    Carly Galek Brian Vincent Grambo Alexander F. Halikias Sarah Hoch Michelle Hochman Iris Hwang Yan Jiang Benjamin Koller Carey Lam Rebecca Susan Lang Shailer E. Lawton Danielle Leavy Jessica Lebitz Melissa Leifer Westhoff Alyssa Ann Wood Yingmei Wu Vincent Yam Seung Mo Yoon Eva Yung Pi Mu Epsilon Pledge I solemnly

  17. Epsilon-Maschinen und Information Bottlenecks

    E-print Network

    Polani, Daniel

    Epsilon-Maschinen und Information Bottlenecks Malte Harder Sommerakademie Olang ­ 17. September statistical complexity. At the other end of the spectrum, simple periodic processes have low statistical on the distinction between K(x) and C(x), another important generalization is to spatially- extended systems -- those

  18. The EPSILON experimental pseudo-symmetric trap

    NASA Astrophysics Data System (ADS)

    Arsenin, V. V.; Dlougach, E. D.; Kulygin, V. M.; Kuyanov, A. Yu.; Skovoroda, A. A.; Timofeev, A. V.; Zhil'tsov, V. A.; Zvonkov, A. V.

    2001-07-01

    Within the framework of the Adaptive Plasma Experiment (APEX) conceptual project, a trap with closed magnetic field lines, the Experimental Pseudo-Symmetric Closed Trap (EPSILON), is examined. The APEX project is aimed at theoretical and experimental development of the physical foundations for a steady state thermonuclear reactor designed on the basis of an alternative magnetic trap with tokamak-like large ? plasma confinement. A discussion is given of the fundamental principle of pseudo-symmetry, which a magnetic configuration with tokamak-like plasma confinement should satisfy. Examples are given of calculations in the paraxial approximation of pseudo-symmetric curvilinear elements with a poloidal modulus B isoline. The EPSILON trap, consisting of two direct axisymmetric mirrors linked by two curvilinear pseudo-symmetric elements, is considered. To increase the equilibrium ?, the plasma currents are short-circuited within curvilinear equilibrium elements. An untraditional scheme of MHD stabilization for a trap with closed field lines by use of axisymmetric mirrors with a divertor is analysed. The experimental installation EPSILON-One Mirror Element (OME), which is under construction for experimental investigation of stabilization by divertor, is discussed. The opportunity for applying the ECR method of plasma production in EPSILON-OME in conditions of high density and low magnetic field is examined.

  19. A Case of Infant Botulism due to Neurotoxigenic Clostridium butyricum Type E Associated with Clostridium difficile Colitis

    Microsoft Academic Search

    L. Fenicia; L. Da Dalt; F. Anniballi; G. Franciosa; S. Zanconato; P. Aureli

    2002-01-01

    .   Reported here is the sixth case of intestinal toxemia botulism caused by Clostridium butyricum type E in Italy since 1984. In this case, the patient was concomitantly affected with colitis due to Clostridium difficile toxin. A review of previously reported cases revealed that some of these patients may also have had intestinal toxemia botulism\\u000a associated with Clostridium difficile colitis,

  20. Rejection of Clostridium putrificum and Conservation of Clostridium botulinum and Clostn'dium sporogenes Request for an Opinion

    Microsoft Academic Search

    INGAR OLSEN; JOHN L. JOHNSON; L. V. H. MOORE; W. E. C. MOORE

    Clostridium putr@cum (Trevisan 1889) Reddish and Rettger 1922; Clostridium botulinum (van Ermengem 1896) Bergey, Harrison, Breed, Hammer, and Huntoon 1923; and Clostridium sporogenes (Mechnikoff 1908) Bergey, Harrison, Breed, Hammer, and Huntoon 1923 are genetically related at the species level. We propose rejection of the name C. putrijicum (which has priority) and conservation of the name C. botulinum on the basis

  1. Pi Mu Epsilon New York Alpha Chapter Membership Application

    E-print Network

    McConnell, Terry

    Pi Mu Epsilon New York Alpha Chapter Membership Application (Due by November 21, 2014) Contact the following questions thoughtfully and specifically on a separate page. 1) What, in your opinion, should Pi Mu Epsilon focus on next year as an organization? 2) In what ways can Pi Mu Epsilon foster a good environment

  2. Applications of Epsilon Radial Networks in Neuroimage Analyses

    E-print Network

    Chung, Moo K.

    Applications of Epsilon Radial Networks in Neuroimage Analyses Nagesh Adluru1, , Moo K. Chung1 using tractography and epsilon neighborhoods were proposed in the diffusion tensor imaging (DTI of such epsilon radial networks (ERN) in performing various types of neu- roimage analyses. These extensions allow

  3. Clostridium difficile in the ICU

    PubMed Central

    Dubberke, Erik R.; Kollef, Marin

    2011-01-01

    Clostridium difficile infection (CDI) management has become more daunting over the past decade because of alarming increases in CDI incidence and severity both in the hospital and in the community. This increase has concomitantly caused significant escalation of the health-care economic burden caused by CDI, and it will likely be translated to increased ICU admission and attributable mortality. Some possible causes for difficulty in management of CDI are as follows: (1) inability to predict and prevent development of severe/complicated or relapsing CDI in patients who initially present with mild symptoms; (2) lack of a method to determine who would have benefited a priori from initiating vancomycin treatment first instead of treatment with metronidazole; (3) lack of sensitive and specific CDI diagnostics; (4) changing epidemiology of CDI, including the emergence of a hypervirulent, epidemic C difficile strain associated with increased morbidity and mortality; (5) association of certain high-usage nonantimicrobial medications with CDI; and (6) lack of treatment regimens that leave the normal intestinal flora undisturbed while treating the primary infection. The objective of this article is to present current management and prevention guidelines for CDI based on recommendations by the Society for Healthcare Epidemiology of America and Infectious Diseases Society of America and potential new clinical management strategies on the horizon. PMID:22147824

  4. Epidemiology of Clostridium difficile Infection

    PubMed Central

    DePestel, Daryl D.; Aronoff, David M.

    2014-01-01

    There has been dramatic change in the epidemiology of Clostridium difficile infection (CDI) since the turn of the 21st Century noted by a marked increase in incidence and severity, occurring at a disproportionately higher frequency in older patients. Historically considered a nosocomial infection associated with antibiotic exposure, CDI has now also emerged in the community in populations previously considered low risk. Emerging risk factors and disease recurrence represent continued challenges in the management of CDI. The increased incidence and severity associated with CDI has coincided with the emergence and rapid spread of a previously rare strain, ribotype 027. Recent data from the U.S. and Europe suggest the incidence of CDI may have reached a crescendo in recent years and is perhaps beginning to plateau. The acute-care direct costs of CDI were estimated to be $4.8 billion in 2008. However, nearly all the published studies have focused on CDI diagnosed and treated in acute-care hospital setting and fail to measure the burden outside the hospital, including recently discharged patients, outpatients, and those in long-term care facilities. Enhanced surveillance methods are needed to monitor the incidence, identify populations at risk, and characterize the molecular epidemiology of strains causing CDI. PMID:24064435

  5. Interactions of high-affinity cationic blockers with the translocation pores of B. anthracis, C. botulinum, and C. perfringens binary toxins.

    PubMed

    Bezrukov, Sergey M; Liu, Xian; Karginov, Vladimir A; Wein, Alexander N; Leppla, Stephen H; Popoff, Michel R; Barth, Holger; Nestorovich, Ekaterina M

    2012-09-19

    Cationic ?-cyclodextrin derivatives were recently introduced as highly effective, potentially universal blockers of three binary bacterial toxins: anthrax toxin of Bacillus anthracis, C2 toxin of Clostridium botulinum, and iota toxin of Clostridium perfringens. The binary toxins are made of two separate components: the enzymatic A component, which acts on certain intracellular targets, and the binding/translocation B component, which forms oligomeric channels in the target cell membrane. Here we studied the voltage and salt dependence of the rate constants of binding and dissociation reactions of two structurally different ?-cyclodextrins (AmPr?CD and AMBnT?CD) in the PA(63), C2IIa, and Ib channels (B components of anthrax, C2, and iota toxins, respectively). With all three channels, the blocker carrying extra hydrophobic aromatic groups on the thio-alkyl linkers of positively charged amino groups, AMBnT?CD, demonstrated significantly stronger binding compared with AmPr?CD. This effect is seen as an increased residence time of the blocker in the channels, whereas the time between blockages characterizing the binding reaction on-rate stays practically unchanged. Surprisingly, the voltage sensitivity, expressed as a slope of the logarithm of the blocker residence time as a function of voltage, turned out to be practically the same for all six cases studied, suggesting structural similarities among the three channels. Also, the more-effective AMBnT?CD blocker shows weaker salt dependence of the binding and dissociation rate constants compared with AmPr?CD. By estimating the relative contributions of the applied transmembrane field, long-range Coulomb, and salt-concentration-independent, short-range forces, we found that the latter represent the leading interaction, which accounts for the high efficiency of blockage. In a search for the putative groups in the channel lumen that are responsible for the short-range forces, we performed measurements with the F427A mutant of PA(63), which lacks the functionally important phenylalanine clamp. We found that the on-rates of the blockage were virtually conserved, but the residence times and, correspondingly, the binding constants dropped by more than an order of magnitude, which also reduced the difference between the efficiencies of the two blockers. PMID:22995493

  6. Identification of Clostridium botulinum, Clostridium argentinense, and related organisms by cellular fatty acid analysis.

    PubMed Central

    Ghanem, F M; Ridpath, A C; Moore, W E; Moore, L V

    1991-01-01

    On the basis of 686 analyses of 285 strains of Clostridium botulinum, Clostridium argentinense (formerly C. botulinum type G), and phenotypically related organisms, 14 cellular fatty acid (CFA) groups of toxic organisms and 6 CFA groups of nontoxic organisms were delineated. The CFA groups of toxic strains included two of type A, three of proteolytic strains of type B, two of proteolytic strains of type F, one each of nonproteolytic strains of types B, E, and F, and one each of types C alpha, C beta, and D and C. argentinense. The groups of phenotypically similar nontoxic strains included Clostridium sporogenes, Clostridium putrificum, nontoxic strains with phenotypic characteristics similar to those of nonproteolytic strains of C. botulinum types B, E, and F (BEF-like), two groups of nontoxigenic organisms with phenotypic characteristics similar to those of C. botulinum types C and D and Clostridium novyi (CDN-like), and Clostridium subterminale, which has phenotypic characteristics similar to those of C. argentinense. Within the toxin types, 89 to 100% of the strains were correctly identified by CFA analysis, and 74 to 100% of the analyses were correct. Of 36 strains of C. sporogenes, 30 (83%) were correctly identified; 17% of the strains of C. sporogenes were incorrectly identified as C. botulinum type A or B. All analyses of C. putrificum and C. subterminale were correctly identified. There was no significant level of similarity between strains of C. botulinum and phenotypically similar organisms and 85 other species of clostridia or 407 other taxa of gram-positive and gram-negative bacteria. Additionally, the one strain each of Clostridium baratii and Clostridium butyricum previously reported to produce C. botulinum toxin could be differentiated from C.botulinum types as well as from strains of C. baratii and C. butyricum that did not produce neurotoxin. PMID:1864927

  7. Secretion of clostridium cellulase by E. coli

    Microsoft Academic Search

    Ida Kuo

    1998-01-01

    A gene, encoding an endocellulase from a newly isolated mesophilic Clostridium strain IY-2 which can digest bamboo fibers, cellulose, rice straw, and sawdust, was isolated by shotgun cloning in an E. coli expression plasmid pLC2833. E. coli positive clones were selected based on their ability to hydrolyze milled bamboo fibers and cellulose present in agar plates. One clone contained a

  8. Clostridium acidurici Electron-Bifurcating Formate Dehydrogenase

    PubMed Central

    Wang, Shuning; Huang, Haiyan; Kahnt, Jörg

    2013-01-01

    Cell extracts of uric acid-grown Clostridium acidurici catalyzed the coupled reduction of NAD+ and ferredoxin with formate at a specific activity of 1.3 U/mg. The enzyme complex catalyzing the electron-bifurcating reaction was purified 130-fold and found to be composed of four subunits encoded by the gene cluster hylCBA-fdhF2. PMID:23872566

  9. Oncolysis by a new strain of clostridium

    Microsoft Academic Search

    U. Mohr; W. Hondius Boldingh; A. Emminger; H Behagel

    1972-01-01

    SUMMARY The oncolytic properties of a new Clostridium strain 01 originating from pig skin extracts have been studied. The specificity toward various animal tumors of a typical pig skin fraction, spore-containing punctates of lysing tumors, and pure spore suspensions proved to be comparable. Several different kinds of transplanted tumors can be destroyed. The activity against spontaneous or induced tumors is

  10. Coculture Production of Butanol by Clostridium Bacteria

    NASA Technical Reports Server (NTRS)

    Bergstrom, S. L.; Foutch, G. L.

    1985-01-01

    Production of butanol by anaerobic fermentation of sugars enhanced by use of two Clostridium species, one of which feeds on metabolic product of other. Renewed interest in fermentation process for making butanol stimulated by potential use of butanol as surfactant in enhanced oil recovery. Butanol also used as fuel or as chemical feedstock and currently produced synthetically from petroleum.

  11. Latest results on the direct CP violation measurements: {Epsilon}{prime}/{epsilon}

    SciTech Connect

    Hsiung, Yee B.; E731 Collaboration

    1992-03-01

    Preliminary results, based on the full data sample from Fermilab-E731 and the combined data sample from CERN-NA31, on the ``direct`` CP-violation measurements {epsilon}{prime}{epsilon} in neutral kaon decay have been reviewed. The E731`s results if Re({epsilon}{prime}{epsilon}) = (6.0 {plus_minus} 5.8(stat) {plus_minus} 3.2(syst) {plus_minus} 1.8(MC)) {times} 10{sup {minus}4}, which provides no evidence for ``direct`` CP-violation, thus supporting the Superweak model; while the NA31`s combined result (1986 + 1988 + 1989 data) is Re({epsilon}{prime}/{epsilon} = (23{plus_minus}3.4(stat) {plus_minus}6.5(syst)){times}10{sup 4}, three standard deviations from zero, which provides evidence for the ``direct`` CP-violation in the Standard Model. Comparisons of the two experiments are made. The Fermilab-E731 group has also fit for the other parameters of the neutral kaon system in their 2{pi} data sample, such as: the K{sub S} life time {tau}{sub S}; the K{sub L}-K{sub S} mass difference {delta}m; the phase difference between {eta}{sub +-}, {delta}{phi} = ({minus}0.6{plus_minus}1.6){degrees}). Which is a test of CPT invariance; the Superweak phase {phi}{sub SW} = (43.37{plus_minus}0.22){degrees}, and the phase of {eta}{sub {plus_minus}}, {phi}{sub {plus_minus}} = (43.2{plus_minus} 1.6){degrees}, which is predicted by CPT invariance to equal {phi}{sub SW}.

  12. Epsilon Aurigae at the End of Eclipse

    NASA Astrophysics Data System (ADS)

    Hoard, Donald; Stencel, R.; Howell, S.

    2011-05-01

    We request a small investment of 24 minutes of Spitzer time, to obtain four IRAC observations of epsilon Aurigae. A naked eye object located near Capella, epsilon Aurigae is the eclipsing binary star with the longest known orbital period, showing a single long duration (~2 yr) eclipse every 27.1 yr. For much of the last 150 years, the nature of the eclipsing object defied explanation. We recently demonstrated that epsilon Aurigae consists of a high luminosity F0 post-AGB star in orbit with a B5 V star surrounded by a solar system sized (~8 AU diameter) disk of cool, dust-dominated material. The eclipse of epsilon Aurigae is a rare event; moreover, it is a unique astrophysical opportunity, since the backlighting of the disk by the high luminosity eclipsed star reveals details that cannot be detected in similar dusty disks around single stars. The current eclipse started in August 2009 and is expected to reach its photometric conclusion in May 2011 (with the spectroscopic conclusion as late as December 2011). The goals for these observations include: (1) extend our ongoing IRAC monitoring campaign covering the current eclipse to late-phase and post-eclipse visits; (2) provide a consistent, well-calibrated space-based set of IR photometry for comparison with ongoing ground-based work; and (3) use the composite results to constrain the thermal profile of the disk. A key expectation of these particular observations is to reveal the irradiation-heated portion of the disk, which will be visible on its trailing side following eclipse. Observations of this side of the disk will be crucial to test and constrain new models of disk structure. As part of our overall monitoring campaign with Spitzer, Hubble, Herschel, and numerous ground-based facilities, these proposed observations will make an important contribution to the understanding of stellar evolution in binary stars, including mass transfer and evolution studies, along with new insights into astrophysical disks and post-AGB star evolution.

  13. Revealing the Hot Side of Epsilon Aurigae

    NASA Astrophysics Data System (ADS)

    Hoard, Donald; Stencel, Robert; Howell, Steve

    2012-12-01

    We request a small investment of 24 minutes of Spitzer time, to obtain four IRAC observations of epsilon Aurigae. A naked eye object located near Capella, epsilon Aurigae is the eclipsing binary star with the longest known orbital period, showing a single long duration (~2 yr) eclipse every 27.1 yr. For much of the last 200 years, the nature of the eclipsing object defied explanation. We recently demonstrated that epsilon Aurigae consists of a high luminosity F0 post-AGB star in orbit with a B5 V star surrounded by a solar system sized (~8 AU diameter) disk of cool, dust-dominated material. The eclipse of epsilon Aurigae is a rare event; moreover, it is a unique astrophysical opportunity, since the backlighting of the disk by the high luminosity eclipsed star reveals details that cannot be detected in similar dusty disks around single stars. The current eclipse started in August 2009 and ended in July 2011; we are now in the post-eclipse phase, when the irradiation-heated side of the disk will begin rotating into view. The goals for these observations include: (1) extend our ongoing IRAC monitoring campaign covering the current eclipse to post-eclipse visits; (2) provide a consistent, well-calibrated space-based set of IR photometry for comparison with ongoing ground-based work; and (3) use the composite results to constrain the thermal profile of the disk. A key expectation of these particular observations is to reveal the irradiation-heated portion of the disk, which will be visible on its trailing side following eclipse. Observations of this side of the disk will be crucial to test and constrain new models of disk structure. As part of our overall monitoring campaign with Spitzer, Hubble, Herschel, and numerous ground-based facilities, these proposed observations will make an important contribution to the understanding of stellar evolution in binary stars, including mass transfer and evolution studies, along with new insights into astrophysical disks and post-AGB star evolution.

  14. The Final Measurement of Epsilon'/Epsilon from KTeV

    SciTech Connect

    Worcester, E.T.

    2009-09-01

    We present precise measurements of CP and CPT symmetry based on the full dataset of K {yields} {pi}{pi} decays collected by the KTeV experiment at FNAL. We measure the direct CP violation parameter Re({epsilon}{prime}/{epsilon}) = (19.2 {+-} 2.1) x 10{sup -4}. We find the KL-KS mass difference {Delta}m = (5265 {+-} 10) x 10{sup 6} hs{sup -1} and the K{sub S} lifetime {tau}{sub S} = (89.62 {+-} 0.05) x 10{sup -12} s. We test CPT symmetry by finding the phase of the indirect CP violation parameter {epsilon}, {phi}{sub {epsilon}} = (44.09 {+-} 1.00){sup o}, and the difference of the relative phases between the CP violating and CP conserving decay amplitudes for K {yields} {pi}{sup +}{pi}{sup -} ({phi}{sub {+-}}) and for K {yields} {pi}{sup 0}{pi}{sup 0} ({phi}{sub 00}), {Delta}{phi} = (0.29 {+-} 0.31){sup o}. These results are consistent with other experimental results and with CPT symmetry.

  15. Identification of Clostridium difficile using the API ZYM system

    Microsoft Academic Search

    P. N. Levett

    1985-01-01

    The use of the API ZYM system for the identification ofClostridium difficile was investigated. The enzyme profiles generated by this system readily distinguished strains ofClostridium difficile from other clostridia commonly isolated from faeces. Enzyme activity ofClostridium difficile was influenced by the composition of the culture medium but appeared to be independent of the age of the culture. Given careful standardisation

  16. Characterization of crotonate grown Clostridium kluyveri by its assimilatory metabolism

    Microsoft Academic Search

    Rudolf K. Thauer; Kurt Jungermann; Joseph Wenning; Karl Decker

    1968-01-01

    Considerable behavioral differences were observed during growth of Clostridium kluyveri on ethanol-acetate and on crotonate media. The identity of the crotonate grown Clostridium with the ethanol grown Clostridium kluyveri was therefore established by three characteristic biosynthetic routes: 1. ribose is synthesized from CO2 and acetate via pyruvate, triose phosphate and a non-oxidative pentose phosphate pathway, 2. reduced one-carbon units are

  17. Recent UBVJH Photometry of Epsilon Aurigae

    E-print Network

    Jeffrey L. Hopkins; Robert E. Stencel

    2007-06-06

    Since first observed in the early 1980s, the Hopkins Phoenix Observatory continues its UBV band observations of the long period (27.1 years) eclipsing binary star system epsilon Aurigae. The UBV observations routinely produce standard deviations or data spread better than 0.01 magnitudes many times approaching 0.001 magnitudes. A new infrared photometer has allowed the addition of near-infrared observations for the JH bands. Typical near-infrared observations approach a standard deviation of data spread of 0.01 magnitudes. The 2003 - 2005 seasons (Fall through Spring) of epsilon Aurigae observations showed a 66.2 day variation that gradually increases in average and peak magnitude in the UBV bands. The 2006 season (Fall 2006 to Spring 2007) data show what appears to be a fall-back to a quiet period near maximum amplitude of V= 3.00. This paper presents the data and compares the current season to the past several seasons. The next eclipse is predicted to begin in 2009 and an international campaign has been organized to coordinate new observations. These website links are: [http://www. hposoft.com/Campaign09.html ] and [ http://www.du.edu/~rstencel/epsaur.htm ] .

  18. Analysis of the Botulinum Neurotoxin Type F Gene Clusters in Proteolytic and Nonproteolytic Clostridium botulinum and Clostridium barati

    Microsoft Academic Search

    Alison K. East; Manju Bhandari; Sebastian Hielm; Matthew D. Collins

    1998-01-01

    .   Comparison of genes encoding type F botulinum neurotoxin progenitor complex in strains of proteolytic Clostridium botulinum strain Langeland, nonproteolytic Clostridium botulinum strain 202F, and Clostridium barati strain ATCC 43256 reveals an identical organization of genes encoding a protein of molecular mass of approx. 47 kDa (P-47),\\u000a nontoxic-nonhemagglutinin (NTNH) and botulinum toxin (BoNT). Although homology between the protein components of

  19. Identification ofClostridium botulinum, Clostridium argentinense, andRelated Organisms byCellular Fatty AcidAnalysis

    Microsoft Academic Search

    FATMA M. GHANEM; C. RIDPATH; W. E. C. MOORE; L. V. H. MOORE

    1991-01-01

    Onthebasis of686analyses of285strains ofClostridium botulinum, Clostridium argentinense (formerly C. botulinum typeG),andphenotypically related organisms, 14cellular fatty acid(CFA)groupsoftoxic organisms and6CFAgroups ofnontoxic organisms weredelineated. TheCFAgroups oftoxic strains included twooftypeA,threeofproteolytic strains oftypeB,twoofproteolytic strains oftypeF,oneeachof nonproteolytic strains oftypes B,E,andF,andoneeachoftypes Ca,CoI, andD andC.argentinense. The groups ofphenotypically similar nontoxic strains included Clostridium sporogenes, Clostridium putrificum, nontoxic strains withphenotypic characteristics similar tothose ofnonproteolytic strains ofC.botulinum types B,E,andF(BEF-like), twogroups

  20. Pi Mu Epsilon Initiation Ceremony Welcome to the second annual induction ceremony of the

    E-print Network

    Feingold, Alex

    Pi Mu Epsilon Initiation Ceremony Welcome to the second annual induction ceremony of the New York Alpha Omicron Chapter of Pi Mu Epsilon, the National Mathematics Honor Society. Information about Pi Mu Epsilon (Janine Belsky): We are assembled here to initiate new members into Pi Mu Epsilon. Pi Mu Epsilon

  1. Biology and Genomic Analysis of Clostridium botulinum

    Microsoft Academic Search

    Michael W. Peck

    2009-01-01

    The ability to form botulinum neurotoxin is restricted to six phylogenetically and physiologically distinct bacteria (Clostridium botulinum Groups I–IV and some strains of C. baratii and C. butyricum). The botulinum neurotoxin is the most potent toxin known, with as little as 30–100ng potentially fatal, and is responsible for botulism, a severe neuroparalytic disease that affects humans, animals, and birds. In

  2. Current advances related to Clostridium difficile infection

    PubMed Central

    Kim, Yong Gil; Jang, Byung Ik

    2015-01-01

    Clostridium difficile infection (CDI) can trigger various responses, ranging from asymptomatic carriage to fulminant colitis. Hard-to-cure CDI, such as severe CDI, multiple recurrences of CDI, refractory CDI, and hypervirulent strains of C. difficile, require new treatments, although antibiotics such as metronidazole and vancomycin are the treatment of choice for initial and first relapsing CDI. Active immunization with C. difficile toxins and faecal microbiota transplantation deserve special attention. Here we describe these strategies for difficult-to-treat CDI. PMID:25900952

  3. Clostridium septicum infection and hemolytic uremic syndrome.

    PubMed Central

    Barnham, M.; Weightman, N.

    1998-01-01

    Five cases of Clostridium septicum infection secondary to Escherichia coli O157-induced hemolytic uremic syndrome have been reported. We report on three cases (one of which is included in the above five) of dual Cl. septicum and E. coil infection; all three patients were exposed to farm animals. A common zoonotic source for Cl. septicum and E. coli O157 infections should be considered. Patients with hemolytic uremic syndrome should be treated aggressively and monitored closely for Cl. septicum superinfection. PMID:9621207

  4. Colonization versus carriage of Clostridium difficile.

    PubMed

    Donskey, Curtis J; Kundrapu, Sirisha; Deshpande, Abhishek

    2015-03-01

    Asymptomatic carriage of toxigenic strains of Clostridium difficile is common in health care facilities and the community. However, infection control efforts have traditionally focused almost entirely on symptomatic patients. There is now growing concern that asymptomatic carriers may be an underappreciated source of transmission. This article provides an overview of the pathogenesis and epidemiology of C difficile colonization, reviews the evidence that asymptomatic carriers shed spores and contribute to transmission, and examines practical issues related to prevention of transmission from carriers. PMID:25595843

  5. Persistent and Recurrent Clostridium difficile Colitis.

    PubMed

    Cole, Shola A; Stahl, Thomas J

    2015-06-01

    Clostridium difficile infection (CDI) is the most frequent cause of nosocomial diarrhea. It has become a significant dilemma in the treatment of patients, and causes increasing morbidity that, in extreme cases, may result in death. Persistent and recurrent disease hamper attempts at eradication of this infection. Escalating levels of treatment and novel therapeutics are being utilized and developed to treat CDI. Further trials are warranted to definitively determine what protocols can be used to treat persistent and recurrent disease. PMID:26034401

  6. Environmental interventions to control Clostridium difficile.

    PubMed

    Loo, Vivian G

    2015-03-01

    The control of Clostridium difficile infection is paramount. C difficile spores are difficult to eradicate and can survive on surfaces for prolonged periods of time. Hand washing with either plain or antimicrobial soap is effective in removing C difficile spores from hands. Patients should be placed in private rooms and under contact precautions to prevent transmission to other patients. Regular hospital germicides are not sporicidal and hypochlorite solutions are required for surface disinfection. In outbreak situations, a multifaceted approach is required. PMID:25573675

  7. Evolving concepts in Clostridium difficile colitis

    Microsoft Academic Search

    Naomi G. Diggs; Christina M. Surawicz

    2009-01-01

    Clostridium difficile infection (CDI) is the most important cause of nosocomial diarrhea. The emergence of a hypervirulent strain and other factors\\u000a including antibiotic overuse contribute to the increasing incidence and severity of this potentially lethal infection. CDI\\u000a has been reported in persons previously considered as low risk, such as young healthy persons without exposure to health care\\u000a settings or antibiotics,

  8. Simulation of turbulence with the k-epsilon model

    Microsoft Academic Search

    B. Cardot; F. Coron; B. Mohammadi; O. Pironneau

    1991-01-01

    The k-epsilon model is investigated for the simulation of turbulence for compressible and incompressible flows. The model is studied from the point of view of the numerical analyst (positivity of k and epsilon, boundedness, etc.). A robust numerical implementation is proposed and several numerical examples are given.

  9. EPSILON SURGERY THEORY STEVE FERRY AND ERIK KJAER PEDERSEN

    E-print Network

    Pedersen, Erik Kjær

    EPSILON SURGERY THEORY STEVE FERRY AND ERIK) by a normal cobordism to obtain a homotopy equiva* *lence f0 : M0 ! X. It is desirable to have an epsilon or controlled version of surgery theory. T* *hus, X comes equipped with a reference map to a metric space K

  10. A k-epsilon-gamma equation turbulence model

    Microsoft Academic Search

    Ji R. Cho; Myung K. Chung

    1992-01-01

    By considering the entrainment effect on the intermittency in the free boundary of shear layers, a set of turbulence model equations for the turbulent kinetic energy k, the dissipation rate epsilon, and the intermittency factor gamma is proposed. This makes it possible to incorporate explicitly the intermittency effect in the conventional k-epsilon turbulence model equations. The eddy viscosity nu(t) is

  11. T*{sub {epsilon}} integral analysis of fracture specimens

    SciTech Connect

    Omori, Y.; Ma, L.; Kobayashi, A.S. [Univ. of Washington, Seattle, WA (United States)] [and others

    1996-12-31

    T*{sub {epsilon}} integral values associated with stable crack growth in thin 2024-T3 aluminum compact (CT) specimens and A606 HSLA steel single edge notched (SEN) specimens were determined directly from the crack tip displacement field obtained by moire interferometry. Stable crack growth in the SEN specimen was also simulated by an elastic-plastic finite element (FE) model which was driven by the experimentally determined boundary conditions. T*{sub {epsilon}} obtained experimentally and by FE were in reasonable agreements with each other. Unlike the vanishing J integrals with crack extension, T*{sub {epsilon}} reached steady state values with stable crack growth. Thus, for a given integration contour, {Gamma}{sub {epsilon}}, near the crack tip, T*{sub {epsilon}} can be used as a stable crack growth as well as a ductile fracture criteria.

  12. Charged Kaon K^+ --> 3 pi CP Violating Asymmetries vs epsilon'_K / epsilon_K

    E-print Network

    Joaquim Prades; Elvira Gamiz; Ignazio Scimemi

    2005-09-29

    We present the next-to-leading order full results in Chiral Perturbation Theory for the charged Kaon K --> 3 pi slope g CP violating asymmetries. We discuss the constraints that a measurement of these asymmetries would impose on the Standard Model results of epsilon_K' and search for new physics. We also study the kind of information that such measurement can provide on Im G_8, Im (e^2 G_E) and higher order weak couplings.

  13. Interferon alpha induces protein kinase C-epsilon (PKC-epsilon) gene expression and a 4.7-kb PKC-epsilon-related transcript.

    PubMed Central

    Wang, C; Constantinescu, S N; MacEwan, D J; Strulovici, B; Dekker, L V; Parker, P J; Pfeffer, L M

    1993-01-01

    Protein kinases play key roles in the induction by human interferon alpha (IFN-alpha) of specific gene expression and biological activity in various human cell lines. We now report that IFN-alpha increased the 7-kb transcript for the epsilon isotype of protein kinase C (PKC-epsilon) and the cellular content of PKC-epsilon 24 and 48 hr after IFN-alpha addition (a 2-fold and 6-fold increase, respectively). Furthermore, IFN-alpha markedly induced a 4.7-kb transcript that hybridized to a PKC-epsilon-specific, but not to a PKC-eta-specific, cDNA probe. The induction of the 4.7-kb PKC-epsilon-related mRNA by IFN-alpha had the following properties reported for the classical IFN-alpha-stimulated genes: rapid kinetics of induction, high maintained levels in IFN-alpha-sensitive but not in IFN-alpha-resistant cell lines, protein synthesis-independent induction, and high sensitivity to inhibitors of protein tyrosine kinase activity. These results show that the regulation of gene expression by IFN-alpha include not only the classical IFN-alpha-stimulated genes but also the coordinated regulation of two PKC-epsilon-related transcripts that appeared to be highly relevant to the biological actions of IFN-alpha. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:8346200

  14. An Atypical Clostridium Strain Related to the Clostridium botulinum Group III Strain Isolated from a Human Blood Culture

    PubMed Central

    Ruimy, Raymond; Bouchier, Christiane; Faucher, Nathalie; Mazuet, Christelle; Popoff, Michel R.

    2014-01-01

    A nontoxigenic strain isolated from a fatal human case of bacterial sepsis was identified as a Clostridium strain from Clostridium botulinum group III, based on the phenotypic characters and 16S rRNA gene sequence, and was found to be related to the mosaic C. botulinum D/C strain according to a multilocus sequence analysis of 5 housekeeping genes. PMID:24088855

  15. Near-wall k-epsilon turbulence modeling

    NASA Technical Reports Server (NTRS)

    Mansour, N. N.; Kim, J.; Moin, P.

    1987-01-01

    The flow fields from a turbulent channel simulation are used to compute the budgets for the turbulent kinetic energy (k) and its dissipation rate (epsilon). Data from boundary layer simulations are used to analyze the dependence of the eddy-viscosity damping-function on the Reynolds number and the distance from the wall. The computed budgets are used to test existing near-wall turbulence models of the k-epsilon type. It was found that the turbulent transport models should be modified in the vicinity of the wall. It was also found that existing models for the different terms in the epsilon-budget are adequate in the region from the wall, but need modification near the wall. The channel flow is computed using a k-epsilon model with an eddy-viscosity damping function from the data and no damping functions in the epsilon-equation. These computations show that the k-profile can be adequately predicted, but to correctly predict the epsilon-profile, damping functions in the epsilon-equation are needed.

  16. VARIABILITY IN OPTICAL SPECTRA OF {epsilon} ORIONIS

    SciTech Connect

    Thompson, Gregory B. [Department of Physics, Adrian College, Adrian, MI 49221 (United States); Morrison, Nancy D., E-mail: gthompson@adrian.edu, E-mail: nmorris@utnet.utoledo.edu [Ritter Astrophysical Research Center, Department of Physics and Astronomy, University of Toledo, 2801 W. Bancroft, Toledo, OH 43606 (United States)

    2013-04-15

    We present the results of a time series analysis of 130 echelle spectra of {epsilon} Ori (B0 Ia), acquired over seven observing seasons between 1998 and 2006 at Ritter Observatory. The equivalent widths of H{alpha} (net) and He I {lambda}5876 were measured and radial velocities were obtained from the central absorption of He I {lambda}5876. Temporal variance spectra (TVS) revealed significant wind variability in both H{alpha} and He I {lambda}5876. The He I TVS have a double-peaked profile consistent with radial velocity oscillations. A periodicity search was carried out on the equivalent width and radial velocity data, as well as on wavelength-binned spectra. This analysis has revealed several periods in the variability with timescales of two to seven days. Many of these periods exhibit sinusoidal modulation in the associated phase diagrams. Several of these periods were present in both H{alpha} and He I, indicating a possible connection between the wind and the photosphere. Due to the harmonic nature of these periods, stellar pulsations may be the origin of some of the observed variability. Periods on the order of the rotational period were also detected in the He I line in the 1998-1999 season and in both lines during the 2004-2005 season. These periods may indicate rotational modulation due to structure in the wind.

  17. EPSILON AURIGAE: AN IMPROVED SPECTROSCOPIC ORBITAL SOLUTION

    SciTech Connect

    Stefanik, Robert P.; Torres, Guillermo; Lovegrove, Justin; Latham, David W.; Zajac, Joseph [Harvard-Smithsonian Center for Astrophysics (CfA), 60 Garden Street, Cambridge, MA 02138 (United States); Pera, Vivian E. [MIT Lincoln Laboratory, 244 Wood Street, Lexington, MA 02420 (United States); Mazeh, Tsevi [Wise Observatory, Tel Aviv University, Tel Aviv 69978 (Israel)], E-mail: rstefanik@cfa.harvard.edu

    2010-03-15

    A rare eclipse of the mysterious object {epsilon} Aurigae will occur in 2009-2011. We report an updated single-lined spectroscopic solution for the orbit of the primary star based on 20 years of monitoring at the CfA, combined with historical velocity observations dating back to 1897. There are 518 new CfA observations obtained between 1989 and 2009. Two solutions are presented. One uses the velocities outside the eclipse phases together with mid-times of previous eclipses, from photometry dating back to 1842, which provide the strongest constraint on the ephemeris. This yields a period of 9896.0 {+-} 1.6 days (27.0938 {+-} 0.0044 years) with a velocity semi-amplitude of 13.84 {+-} 0.23 km s{sup -1} and an eccentricity of 0.227 {+-} 0.011. The middle of the current ongoing eclipse predicted by this combined fit is JD 2,455,413.8 {+-} 4.8, corresponding to 2010 August 5. If we use only the radial velocities, we find that the predicted middle of the current eclipse is nine months earlier. This would imply that the gravitating companion is not the same as the eclipsing object. Alternatively, the purely spectroscopic solution may be biased by perturbations in the velocities due to the short-period oscillations of the supergiant.

  18. Antibodies for Treatment of Clostridium difficile Infection

    PubMed Central

    Wilcox, Mark H.

    2014-01-01

    Antibodies for the treatment of Clostridium difficile infection (CDI) have been demonstrated to be effective in the research and clinical environments. Early uncertainties about molecular and treatment modalities now appear to have converged upon the systemic dosing of mixtures of human IgG1. Although multiple examples of high-potency monoclonal antibodies (MAbs) exist, significant difficulties were initially encountered in their discovery. This minireview describes historical and contemporary MAbs and highlights differences between the most potent MAbs, which may offer insight into the pathogenesis and treatment of CDI. PMID:24789799

  19. Bacteria associated with processed crawfish and potential toxin production by Clostridium botulinum type E in vacuum-packaged and aerobically packaged crawfish tails.

    PubMed

    Lyon, W J; Reddmann, C S

    2000-12-01

    Refrigerated vacuum-packaged storage has been shown to increase significantly the shelf life of fresh fish and seafood products, but the effect, if any, on the outgrowth and toxin production of Clostridium botulinum type E on cooked crawfish is unknown. Microflora associated with live crawfish reflect the microbial populations of the harvest water and sediments in which they are living. The presence or absence of specific pathogens in either vacuum-packaged or air-permeable bags of cooked crawfish have not been thoroughly evaluated. This study evaluates the potential survival and outgrowth of biological hazards in both vacuum-packaged and air-permeable-packaged cooked crawfish held at 4 and 10 degrees C for 30 days. During shelf-life studies of vacuum-packaged and air-permeable-bagged cooked crawfish, a total of 31 bacterial species were isolated and identified from crawfish samples using both selective and nonselective media. The only pathogens isolated from both vacuum-packed and air-permeable bags of processed crawfish samples during shelf-life studies were strains of Aeromonas hydrophila and Staphylococcus aureus. C. botulinum type E and Clostridium perfringens species were not isolated from any of the uninoculated crawfish samples. Cooked crawfish were inoculated with 10(3) C. botulinum type E spores per g of crawfish tail meat to determine whether cooked crawfish tails would support the growth of C. botulinum type E strains and produce toxin at refrigerated temperatures. Spore-inoculated crawfish tails were vacuum packaged in both a high barrier film and an air-permeable bag and stored at 4 degrees C and 10 degrees C for 30 days. C. botulinum toxin E was not detected in any of the spore-inoculated packages throughout the shelf-life study until day 30. Microbiological data from this study should be useful in the development and implementation of the hazard analysis and critical control point plans for processed crawfish tails. PMID:11131892

  20. Complete Genome Sequence of Clostridium clariflavum DSM 19732

    PubMed Central

    Izquierdo, Javier A.; Goodwin, Lynne; Davenport, Karen W.; Teshima, Hazuki; Bruce, David; Detter, Chris; Tapia, Roxanne; Han, Shunsheng; Land, Miriam; Hauser, Loren; Jeffries, Cynthia D.; Han, James; Pitluck, Sam; Nolan, Matt; Chen, Amy; Huntemann, Marcel; Mavromatis, Konstantinos; Mikhailova, Natalia; Liolios, Konstantinos; Woyke, Tanja; Lynd, Lee R.

    2012-01-01

    Clostridium clariflavum is a Cluster III Clostridium within the family Clostridiaceae isolated from thermophilic anaerobic sludge (Shiratori et al, 2009). This species is of interest because of its similarity to the model cellulolytic organism Clostridium thermocellum and for the ability of environmental isolates to break down cellulose and hemicellulose. Here we describe features of the 4,897,678 bp long genome and its annotation, consisting of 4,131 protein-coding and 98 RNA genes, for the type strain DSM 19732. PMID:22675603

  1. An epsilon wave in arrhythmogenic right ventricular cardiomyopathy\\/dysplasia

    Microsoft Academic Search

    Chun-Chi You; Yuan-Teng Tseng; Ming-Hsiung Hsieh

    2007-01-01

    A 53-year-old male presented to our emergency department with a sudden onset of ventricular tachycardia with left bundle branch block and right axis deviation. After the tachyarrhythmia converting to sinus rhythm, the ECG displayed sinus rhythm with a typical epsilon wave in leads V1 and V2. Arrhythmogenic right ventricular dysplasia\\/cardiomyopathy (ARVD\\/C) was suspected. The epsilon wave is the most specific

  2. An extended k-epsilon finite element model

    Microsoft Academic Search

    Marc Jaeger; Gouri Dhatt

    1992-01-01

    An extended k-epsilon model (to include low-Reynolds-number regions) employing weighting functions is presented. Wall functions for the near-wall zones are developed giving correct boundary values for the shear stress and k-epsilon. A finite element model using a penalty formulation for incompressible turbulent flow is applied to solve a flow between two plates. Results with mesh boundaries situated in the near-wall

  3. Secretion of clostridium cellulase by E. coli

    DOEpatents

    Yu, Ida Kuo (1885 California St., #62, Mountain View, CA 94041)

    1998-01-01

    A gene, encoding an endocellulase from a newly isolated mesophilic Clostridium strain IY-2 which can digest bamboo fibers, cellulose, rice straw, and sawdust, was isolated by shotgun cloning in an E. coli expression plasmid pLC2833. E. coli positive clones were selected based on their ability to hydrolyze milled bamboo fibers and cellulose present in agar plates. One clone contained a 2.8 kb DNA fragment that was responsible for cellulase activity. Western blot analyses indicated that the positive clone produced a secreted cellulase with a mass of about 58,000 daltons that was identical in size to the subunit of one of the three major Clostridium cellulases. The products of cellulose digestion by this cloned cellulase were cellotetraose and soluble higher polymers. The cloned DNA contained signal sequences capable of directing the secretion of heterologous proteins from an E. coli host. The invention describes a bioprocess for the treatment of cellulosic plant materials to produce cellular growth substrates and fermentation end products suitable for production of liquid fuels, solvents, and acids.

  4. Clostridium to treat cancer: dream or reality?

    PubMed Central

    Lambin, Philippe

    2015-01-01

    In their paper “Intratumoral injection of Clostridium novyi-NT spores induces antitumor responses”, Roberts et al. describe the induction of antitumor responses following local spore administration of an attenuated C. novyi strain (C. novyi-NT). Stereotactic intratumoral spore injection led to significant survival advantages in a murine orthotopic brain model and local bacterial treatment produced robust responses in a set of spontaneous canine soft tissue carcinomas. Their preclinical findings in both models, provided the basis for a phase 1 investigational clinical study in patients with solid tumors that were either refractory to standard treatment or without an available standard treatment available (NCT01924689). The results of the first patient enrolled in this trial, a 53-year-old female with a retroperitoneal leiomyosarcoma, are described. Next to the non-armed C. novyi-NT described in this paper, very potent genetically modified Clostridium expressing anti-cancer therapeutic genes are also being developed. Are treatments with these non-pathogenic clostridia a viable alternative cancer treatment?

  5. Development of an antigen-capture enzyme-linked immunosorbent assay for Clostridium perfringens beta2-toxin in porcine feces and the neonatal piglet intestine.

    PubMed

    Kircanski, Jasmina; Hodgins, Douglas; Soltes, Glenn; Pei, Yanlong; Parreira, Valeria R; Songer, J Glenn; Prescott, John F

    2012-09-01

    An enzyme-linked immunosorbent assay (ELISA) was developed for detection and quantitation of beta2-toxin in neonatal piglet intestinal contents. Polystyrene plates were coated with polyclonal capture antibodies prepared against consensus recombinant beta2-toxin. The ELISA was developed using consensus recombinant beta2-toxin, atypical recombinant beta2-toxin, purified consensus native beta2-toxin, and field samples of neonatal porcine intestinal contents. Captured antigen was detected using a horseradish peroxidase-labeled monoclonal antibody against consensus recombinant beta2-toxin. The limit of detection of the ELISA for consensus beta2-toxin was between 2.0 and 3.5 ng/ml. The ELISA detected atypical recombinant beta2-toxin only weakly. Optical density was protein concentration dependent. The test confirmed differences between consensus and atypical recombinant beta2-toxin, but similar results obtained when testing pure consensus recombinant beta2-toxin and native beta2-toxin. Results obtained from intestinal content samples, particularly from the small intestine, were highly inconsistent and suggested variable protease activity. Addition of protease inhibitors partially prevented degradation of the toxin; however, sample processing at low temperature, at a lower pH (citrate buffer with 5% of bovine serum albumin, pH 6.1), and "cold incubation" of applied antigens abolished protease activity. The recombinant toxin was preserved in spiked intestinal samples by freezing at -70°C, suggesting that necropsy samples can be stored frozen for periodic testing. With appropriate sample preparation, antigen-capture ELISA can detect beta2-toxin in the intestinal content and feces of neonatal piglets. PMID:22855377

  6. Humulus lupus Beta-acids Administered Through Water Reduce Clostridium perfringens Challenge Strains in the Chicken Intestinal Tract Midgut and Ceca.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The antimicrobial activity activity of extracts of the hop plant Humulus lupus was studied in chickens fed diets without antibiotic growth promotants. Beta-acid resins of the hop plant were administered by water to 13 day old chickens subsequently challenged per so with necrotic enteritis-associate...

  7. Heat resistance and outgrowth of clostridium perfringens spores as affected by the type of heating medium, and heating and cooling rates in ground pork 

    E-print Network

    Marquez Gonzalez, Mayra

    2009-05-15

    subsequent use. Frozen packages were thawed by submerging a 300 g package in a plastic tub with water at 25?C for 30 min before use. The thawed packaged was dried using paper towels and sanitized with 70% isopropyl alcohol presaturated wipes (SATWipes...

  8. Copolymers of epsilon-caprolactone and quaternized epsilon-caprolactone as gene carriers.

    PubMed

    Vroman, Benoît; Mazza, Michaël; Fernandez, Manuela R; Jérôme, Robert; Préat, Véronique

    2007-03-12

    New copolymers of epsilon-caprolactone (CL) and gamma-bromo-epsilon-caprolactone quaternized by pyridine (Py+CL) were investigated as non-viral vectors for gene delivery. Copolymers with two molar compositions (50 Py+CL/50 CL and 80 Py+CL/20 CL), each with a diblock or a random structure, were used to prepare nanoparticulate complexes with DNA. Average size and surface charge of the complexes and extent of the complexation were measured. The DNA condensation by the copolymers was analysed by a gel retardation assay. Cytotoxicity and transfection efficiency of the copolymers were also evaluated in HeLa cells and compared with polyethylenimine 50 kDa. The size of the polyplexes was approximately 200 nm. The zeta potential first increased with the copolymer/DNA charge ratio and became positive for charge ratios in the 2-4 range depending on the type of copolymer. DNA was completely condensed within the nanoparticles and the degree of interaction was very high. Cytotoxicity and transfection efficiency were found to be comparable to polyethylenimine 50 kDa. The experimental results suggest that the novel copolymers can be used as novel gene delivery vectors. PMID:17258343

  9. Correlations between epsilon'/epsilon and Rare K Decays in the Littlest Higgs Model with T-Parity

    E-print Network

    Monika Blanke; Andrzej J. Buras; Stefan Recksiegel; Cecilia Tarantino; Selma Uhlig

    2010-01-15

    We calculate the CP-violating ratio epsilon'/epsilon in the Littlest Higgs model with T-parity (LHT) and investigate its correlations with the branching ratios for K_L -> pi^0 nu {bar nu}, K_L -> pi^0 l^+ l^- and K^+ -> pi^+ nu {bar nu}. The resulting correlations are rather strong in the case of K_L decays, but less pronounced in the case of K^+ -> pi^+ nu {bar nu}. Unfortunately, they are subject to large hadronic uncertainties present in epsilon'/epsilon, whose theoretical prediction in the Standard Model (SM) is reviewed and updated here. With the matrix elements of Q_6 (gluon penguin) and Q_8 (electroweak penguin) evaluated in the large-N limit and m_s^MS(2 GeV) = 100 MeV from lattice QCD, (epsilon'/epsilon)_SM turns out to be close to the data so that significant departures of Br(K_L -> pi^0 nu {bar nu}) and Br(K_L -> pi^0 l^+ l^-) from the SM expectations are unlikely, while Br(K^+ -> pi^+ nu {bar nu}) can be enhanced even by a factor 5. On the other hand, modest departures of the relevant hadronic matrix elements from their large-N values allow for a consistent description of epsilon'/epsilon within the LHT model accompanied by large enhancements of Br(K_L -> pi^0 nu {bar nu}) and Br(K_L -> pi^0 l^+ l^-), but only modest enhancements of Br(K^+ -> pi^+ nu {bar nu}).

  10. Theoretical estimate(s) of the CP-violating quontity epsilon-prime/epsilon in K -> 2 pi decays

    E-print Network

    J. O. Eeg

    2000-10-05

    I give a short presentation of the theoretical prediction of epsilon-prime/epsilon. Short distance and especially long distance aspects of the computation is discussed. I consentrate on the general framework and the chiral quark model approach, while other approaches are also shortly presented. Because of the intrinsic uncertainties of the long-distance computations, it is unlikely that new physics effects can be disentangled from the standard model prediction.

  11. Crystal structure of the Clostridium limosum C3 exoenzyme

    Microsoft Academic Search

    Martin Vogelsgesang; Benjamin Stieglitz; Christian Herrmann; Alex Pautsch; Klaus Aktories

    2008-01-01

    C3-like toxins ADP-ribosylate and inactivate Rho GTPases. Seven C3-like ADP-ribosyltransferases produced by Clostridium botulinum, Clostridium limosum, Bacillus cereus and Staphylococcus aureus were identified and two representatives – C3bot from C. botulinum and C3stau2 from S. aureus – were crystallized. Here we present the 1.8Å structure of C. limosum C3 transferase C3lim and compare it to the structures of other family

  12. Evidence for quorum sensing in Clostridium botulinum 56A

    Microsoft Academic Search

    L. Zhao; T. J. Montville; D. W. Schaffner

    2006-01-01

    Aims: Experiments were designed to detect quorum-sensing signals produced by Clostridium botulinum. Methods and Results: Clostridium botulinum 56A cell-free supernatants obtained at the end of lag phase, the mid-exponential phase and early station- ary phase of growth were assayed for bioluminescence in the Vibrio harveyi quorum-sensing assay system. Twelve and 16-h culture supernatants induced bioluminescence in the auto-inducer 2 (AI-2)

  13. Arthrite réactionnelle après infection par Clostridium difficile chez deux enfants

    Microsoft Academic Search

    Helga A Löffer; Benedicte Pron; Richard Mouy; Nico M Wulffraat; Anne-Marie Prieur

    2004-01-01

    In adults, reactive arthritis (ReA) following Clostridium difficile-enterocolitis has been documented. In children, only one case of Clostridium difficile (C. difficile) associated ReA in a child has been reported. We now describe 2 other cases of ReA associated with C. difficile in children. The characteristics of ReA due to C. difficile appear to be similar in adults and children. Both

  14. Update on antimicrobial resistance in Clostridium difficile.

    PubMed

    Qiong, Gao; Haihui, Huang

    2015-05-01

    Clostridium difficile is the leading cause of healthcare-associated diarrhea. Since 2002, the morbidity and mortality rates of C. difficile infection have increased dramatically in Europe and North America. The emergence of C. difficile strains that are resistant to multiple antimicrobial agents can complicate prevention programs and potential treatment. Although most clinical isolates are still susceptible to metronidazole and vancomycin, heteroresistance to metronidazole and increasing vancomycin MICs (minimum inhibitory concentrations) have been reported. The prevalence of resistance to other antimicrobial agents, including erythromycin and moxifloxacin, is highly variable in different countries and regions. The exact mechanism of reduced susceptibility to metronidazole or vancomycin is still not clear. The principal mechanism of erythromycin, fluoroquinolones and rifamycins resistance in C. difficile is determined by target alterations. This review will focus primarily on the antimicrobial susceptibility patterns and resistance mechanisms of C. difficile in order to provide an up-to-date review on the topic. PMID:25998434

  15. Clostridium difficile: clinical disease and diagnosis.

    PubMed Central

    Knoop, F C; Owens, M; Crocker, I C

    1993-01-01

    Clostridium difficile is an opportunistic pathogen that causes a spectrum of disease ranging from antibiotic-associated diarrhea to pseudomembranous colitis. Although the disease was first described in 1893, the etiologic agent was not isolated and identified until 1978. Since clinical and pathological features of C. difficile-associated disease are not easily distinguished from those of other gastrointestinal diseases, including ulcerative colitis, chronic inflammatory bowel disease, and Crohn's disease, diagnostic methods have relied on either isolation and identification of the microorganism or direct detection of bacterial antigens or toxins in stool specimens. The current review focuses on the sensitivity, specificity, and practical use of several diagnostic tests, including methods for culture of the etiologic agent, cellular cytotoxicity assays, latex agglutination tests, enzyme immunoassay systems, counterimmunoelectrophoresis, fluorescent-antibody assays, and polymerase chain reactions. PMID:8358706

  16. Clostridium difficile infection in older adults

    PubMed Central

    Jump, Robin LP

    2014-01-01

    Clostridium difficile infection, the most frequent cause of nosocomial diarrhea, disproportionately affects older adults. The two most important risk factors for developing C. difficile infection are antimicrobial exposure and age >65 years old. Risk factors specific to older adults are frequent interactions with healthcare systems and age-related changes in physiology, including immune senescence and changes to the gut microbiome. Metronidazole and oral vancomcyin are the mainstays of conventional treatment for C. difficile infection. Alternative therapies include fidaxomicin, a narrow-spectrum macrocyclic antibiotic, and fectal bacteriotherapy, which offers an excellent therapeutic outcome. Strategies to prevent C. difficile infections include enhanced infection control measures and reducing inappropriate antimicrobial use through stewardship. PMID:24955106

  17. Characterization of endoglucanase A from Clostridium cellulolyticum.

    PubMed

    Fierobe, H P; Gaudin, C; Belaich, A; Loutfi, M; Faure, E; Bagnara, C; Baty, D; Belaich, J P

    1991-12-01

    A construction was carried out to obtain a high level of expression in Escherichia coli of the gene celCCA, coding for the endoglucanase A from Clostridium cellulolyticum (EGCCA). The enzyme was purified in two forms with different molecular weights, 51,000 and 44,000. The smaller protein was probably the result of proteolysis, although great care was taken to prevent this process from occurring. Evidence was found for the loss of the conserved reiterated domains which are characteristic of C. thermocellum and C. cellulolyticum cellulases. The two forms were extensively studied, and it was demonstrated that although they had the same pH and temperature optima, they differed in their catalytic properties. The truncated protein gave the more efficient catalytic parameters on carboxymethyl cellulose and showed improved endoglucanase characteristics, whereas the intact enzyme showed truer cellulase characteristics. The possible role of clostridial reiterated domains in the hydrolytic activity toward crystalline cellulose is discussed. PMID:1744052

  18. Characterization of endoglucanase A from Clostridium cellulolyticum.

    PubMed Central

    Fierobe, H P; Gaudin, C; Belaich, A; Loutfi, M; Faure, E; Bagnara, C; Baty, D; Belaich, J P

    1991-01-01

    A construction was carried out to obtain a high level of expression in Escherichia coli of the gene celCCA, coding for the endoglucanase A from Clostridium cellulolyticum (EGCCA). The enzyme was purified in two forms with different molecular weights, 51,000 and 44,000. The smaller protein was probably the result of proteolysis, although great care was taken to prevent this process from occurring. Evidence was found for the loss of the conserved reiterated domains which are characteristic of C. thermocellum and C. cellulolyticum cellulases. The two forms were extensively studied, and it was demonstrated that although they had the same pH and temperature optima, they differed in their catalytic properties. The truncated protein gave the more efficient catalytic parameters on carboxymethyl cellulose and showed improved endoglucanase characteristics, whereas the intact enzyme showed truer cellulase characteristics. The possible role of clostridial reiterated domains in the hydrolytic activity toward crystalline cellulose is discussed. Images FIG. 2 PMID:1744052

  19. Clostridium difficile Infection: New Insights Into Management

    PubMed Central

    Khanna, Sahil; Pardi, Darrell S.

    2012-01-01

    Clostridium difficile was first described as a cause of diarrhea in 1978 and is now among the leading 3 hospital-acquired infections in the United States, along with methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci. In the past 2 decades, there has been an increase in the incidence, severity, and recurrence rates of C difficile infection, all of which are associated with poor outcomes. In addition, several novel risk factors and newer treatment methods are emerging, including fidaxomicin therapy, treatment using monoclonal antibodies, and fecal microbiota transplantation, that have shown promise for the treatment of C difficile infection. This review focuses on the changing epidemiology, risk factors, and newer methods for treatment of C difficile infection. PMID:23127735

  20. Radial velocity study of the CP star Epsilon Ursae Majoris

    E-print Network

    Sokolov, N A

    2009-01-01

    In this Letter, the radial velocity variability of the chemically peculiar star Epsilon Ursae Majoris ($\\epsilon$ UMa) from the sharp cores of the hydrogen lines is investigated. This study is based on the ELODIE archival data obtained at different phases of the rotational cycle. The star exhibits low-amplitude radial velocity variations with a period of P=5.0887 d. The best Keplerian solution yields an eccentricity e=0.503 and a minimum mass ~14.7$M_{\\rm Jup}$ on the hypothesis that the rotational axis of $\\epsilon$ UMa is perpendicular to the orbital plane. This result indicate that the companion is the brown-dwarf with the projected semi-amplitude variation of the radial velocity $K_{\\rm 2}$=135.9 km/sec and the sine of inclination times semi-major axis $a_{2}$sin(i)=0.055 au.

  1. Little Randall-Sundrum models: epsilon_K strikes again

    E-print Network

    M. Bauer; S. Casagrande; L. Gruender; U. Haisch; M. Neubert

    2008-11-22

    A detailed phenomenological analysis of neutral kaon mixing in "little Randall-Sundrum" models is presented. It is shown that the constraints arising from the CP-violating quantity epsilon_K can, depending on the value of the ultra-violet cutoff, be even stronger than in the original Randall-Sundrum scenario addressing the hierarchy problem up to the Planck scale. The origin of the enhancement is explained, and a bound Lambda_{UV}>several 10^3 TeV is derived, below which vast corrections to epsilon_K are generically unavoidable. Implications for non-standard Z->bb couplings are briefly discussed.

  2. Epsilon-Near-Zero Mode for Active Optoelectronic Devices

    NASA Astrophysics Data System (ADS)

    Vassant, S.; Archambault, A.; Marquier, F.; Pardo, F.; Gennser, U.; Cavanna, A.; Pelouard, J. L.; Greffet, J. J.

    2012-12-01

    The electromagnetic modes of a GaAs quantum well between two AlGaAs barriers are studied. At the longitudinal optical phonon frequency, the system supports a phonon polariton mode confined in the thickness of the quantum well that we call epsilon-near-zero mode. This epsilon-near-zero mode can be resonantly excited through a grating resulting in a very large absorption localized in the single quantum well. We show that the reflectivity can be modulated by applying a voltage. This paves the way to a new class of active optoelectronic devices working in the midinfrared and far infrared at ambient temperature.

  3. Phylogeny of the ammonia-producing ruminal bacteria Peptostreptococcus anaerobius, Clostridium sticklandii, and Clostridium aminophilum sp. nov

    NASA Technical Reports Server (NTRS)

    Paster, B. J.; Russell, J. B.; Yang, C. M.; Chow, J. M.; Woese, C. R.; Tanner, R.

    1993-01-01

    In previous studies, gram-positive bacteria which grew rapidly with peptides or an amino acid as the sole energy source were isolated from bovine rumina. Three isolates, strains C, FT (T = type strain), and SR, were considered to be ecologically important since they produced up to 20-fold more ammonia than other ammonia-producing ruminal bacteria. On the basis of phenotypic criteria, the taxonomic position of these new isolates was uncertain. In this study, the 16S rRNA sequences of these isolates and related bacteria were determined to establish the phylogenetic positions of the organisms. The sequences of strains C, FT, and SR and reference strains of Peptostreptococcus anaerobius, Clostridium sticklandii, Clostridium coccoides, Clostridium aminovalericum, Acetomaculum ruminis, Clostridium leptum, Clostridium lituseburense, Clostridium acidiurici, and Clostridium barkeri were determined by using a modified Sanger dideoxy chain termination method. Strain C, a large coccus purported to belong to the genus Peptostreptococcus, was closely related to P. anaerobius, with a level of sequence similarity of 99.6%. Strain SR, a heat-resistant, short, rod-shaped organism, was closely related to C. sticklandii, with a level of sequence similarity of 99.9%. However, strain FT, a heat-resistant, pleomorphic, rod-shaped organism, was only distantly related to some clostridial species and P. anaerobius. On the basis of the sequence data, it was clear that strain FT warranted designation as a separate species. The closest known relative of strain FT was C. coccoides (level of similarity, only 90.6%). Additional strains that are phenotypically similar to strain FT were isolated in this study.(ABSTRACT TRUNCATED AT 250 WORDS).

  4. Inhibition of insulin-stimulated glucose transport in 3T3-L1 cells by Clostridium difficile toxin B, Clostridium sordellii lethal toxin, and Clostridium botulinum C2 toxin

    Microsoft Academic Search

    Gudrun Schmid; Annette Schürmann; Christine Huppertz; Fred Hofmann; Klaus Aktories; Hans-Georg Joost

    1998-01-01

    The role of the actin cytoskeleton and\\/or GTPases of the Rho\\/Rac-family in glucose transport regulation was investigated in\\u000a 3T3-L1 cells with clostridial toxins which depolymerize actin by inactivation of Rho\\/Rac (Clostridium difficile toxin B and Clostridium sordellii lethal toxin (LT)) or by direct ADP-ribosylation (Clostridium botulinum C2 toxin). Toxin B and C2 reduced insulin-stimulated, but not basal, 2-deoxyglucose (2-DOG) uptake

  5. Analysis of the {Delta}I=1/2 rule and {epsilon}'/{epsilon} with overlap fermions

    SciTech Connect

    Capitani, Stefano; Giusti, Leonardo

    2001-07-01

    We study the renormalization of the {Delta}S=1 effective weak Hamiltonian with overlap fermions. The mixing coefficients among dimension-6 operators are computed at one loop in perturbation theory. As a consequence of the chiral symmetry at finite lattice spacing and of the GIM mechanism, which is quadratic in the masses, the K{yields}{pi}{pi} and K{yields}{pi} matrix elements relevant for the {Delta}I=1/2 rule can be computed without any power subtractions. The analogous amplitudes for {epsilon}'/{epsilon} require one divergent subtraction only, which can be performed nonperturbatively using K{yields}0 matrix elements.

  6. Analysis of the {Delta}I=1/2 rule and {epsilon}{prime}/{epsilon} with overlap fermions

    SciTech Connect

    Capitani, Stefano; Giusti, Leonardo

    2001-07-01

    We study the renormalization of the {Delta}S=1 effective weak Hamiltonian with overlap fermions. The mixing coefficients among dimension-6 operators are computed at one loop in perturbation theory. As a consequence of the chiral symmetry at finite lattice spacing and of the GIM mechanism, which is quadratic in the masses, the K{r_arrow}{pi}{pi} and K{r_arrow}{pi} matrix elements relevant for the {Delta}I=1/2 rule can be computed without any power subtractions. The analogous amplitudes for {epsilon}{prime}/{epsilon} require one divergent subtraction only, which can be performed nonperturbatively using K{r_arrow}0 matrix elements.

  7. Clostridium difficile infection in an Iranian hospital

    PubMed Central

    2012-01-01

    Background Clostridium difficile infection (CDI) is an important cause of morbidity and mortality internationally, yet there are important regional differences in the epidemiology and microbiology of disease. Most reports have come from North America and Europe, with limited information from other regions, including the Middle East. Given the changes in the epidemiology of CDI in developed countries, particularly associated with the dissemination of hypervirulent epidemic clones, an understanding of the epidemiology and microbiology of CDI in diverse regions is warranted. This study involved collection of stool samples from individuals with diarrhea at the Isfahan University of Medical Sciences Teaching Hospital, Isfahan, Iran, between October 2010 and March 2011. Selective enrichment culture for C. difficile was performed and isolates were characterised using ribotyping, PCR for the detection of tcdA, tcdB and cdtB genes, and tcdC sequence analysis. Findings Clostridium difficile was isolated from 19/89 (21%) stool samples of 17/86 (20%) patients. 13/17 (77%) cases of CDI were hospital-associated. Patients with CDI were significantly older (43 ± 28y) than those with non-CDI diarrhea (24, ± 26y)(P = 0.018). All isolates were toxigenic, and possessed genes encoding for toxins A and B. Six (32%) of 19 isolates also possessed cdtB. Twelve ribotypes were identified. Ribotype 078/toxinotype V was most common, accounting for 4 (21%) of isolates. A single isolate of a different toxinotype V ribotype was identified, as was a toxinotype XXIV isolate. The remaining isolates consisted of 9 different toxinotype 0 ribotypes. Conclusions CDI is an important cause of diarrhea in patients in this hospital. The diversity of ribotypes was striking, and the number of different types suggests the presence of a broad range of strains in the community, the hospital or both. The predominance of toxinotype V strains, which have been associated with community-associated disease and food animals, was unexpected and possible sources of this type require further investigation. PMID:22436392

  8. Epsilon-Unfolding Orthogonal Polyhedra Mirela Damian1

    E-print Network

    O'Rourke, Joseph

    unfolding of [1], in the spiral strips used in [6], and the band structure exploited in [8], but introducesEpsilon-Unfolding Orthogonal Polyhedra Mirela Damian1 , Robin Flatland2 , Joseph O'Rourke3 1 Dept@cs.smith.edu Abstract. An unfolding of a polyhedron is produced by cutting the surface and flat- tening to a single

  9. An improved k-epsilon model for near wall turbulence

    NASA Technical Reports Server (NTRS)

    Shih, T. H.; Hsu, Andrew T.

    1991-01-01

    An improved k-epsilon model for low Reynolds number turbulence near a wall is presented. In the first part of this work, the near-wall asymptotic behavior of the eddy viscosity and the pressure transport term in the turbulent kinetic energy equation are analyzed. Based on these analyses, a modified eddy viscosity model with the correct near-wall behavior is suggested, and a model for the pressure transport term in the k-equation is proposed. In addition, a modeled dissipation rate equation is reformulated, and a boundary condition for the dissipation rate is suggested. In the second part of the work, one of the deficiencies of the existing k-epsilon models, namely, the wall distance dependency of the equations and the damping functions, is examined. An improved model that does not depend on any wall distance is introduced. Fully developed turbulent channel flows and turbulent boundary layers over a flat plate are studied as validations for the proposed new models. Numerical results obtained from the present and other previous k-epsilon models are compared with data from direct numerical simulation. The results show that the present k-epsilon model, with added robustness, performs as well as or better than other existing models in predicting the behavior of near-wall turbulence.

  10. EPSILON SURGERY THEORY STEVE FERRY AND ERIK KJR PEDERSEN

    E-print Network

    Pedersen, Erik Kjær

    EPSILON SURGERY THEORY STEVE FERRY AND ERIK KJÆR PEDERSEN Contents 1. Introduction 2 2. Algebraic preliminaries 4 3. Bounded Poincar´e complexes 8 4. Spivak normal fibre space 9 5. Surgery below the middle. The surgery groups 22 10. Ranicki-Rothenberg sequences, and L- 26 11. The surgery exact sequence 29 12

  11. Characterization of Anopheles dirus Glutathione Transferase Epsilon 4

    Microsoft Academic Search

    Gulsiri Charoensilp; Ardcharaporn Vararattanavech; Posri Leelapat; La-aied Prapanthadara; Albert J. Ketterman

    2006-01-01

    The coding sequences of a wild type glutathione transferase (GST) Epsilon 4 and three isoenzymes were obtained by RT-PCR from a Thai malaria mosquito, Anopheles dirus. After confirmation by sequencing, the RT-PCR products were subcloned into an expression vector and proteins were expressed, purified, and biochemically characterized to study the function of these enzymes and for comparison with two orthologs

  12. Exploring the epsilon regime with twisted mass fermions

    E-print Network

    K. Jansen; A. Nube; A. Shindler; C. Urbach; U. Wenger

    2007-11-13

    In this proceeding contribution we report on a first study in order to explore the so called epsilon regime with Wilson twisted mass (Wtm) fermions. To show the potential of this approach we give a preliminary determination of the chiral condensate.

  13. Meson Correlation Functions in the epsilon-Regime

    E-print Network

    T. Chiarappa; W. Bietenholz; K. Jansen; K. -I. Nagai; S. Shcheredin

    2003-10-28

    We present a numerical pilot study of the meson correlation functions in the epsilon-regime of chiral perturbation theory. Based on simulations with overlap fermions we measured the axial and pseudo-scalar correlation functions, and we discuss the implications for the leading low energy constants in the chiral Lagrangian.

  14. Simulating chiral quarks in the epsilon-regime of QCD

    E-print Network

    Bietenholz, W; Jansen, K; Nagai, K I; Shcheredin, S

    2005-01-01

    We present simulation results for lattice QCD with chiral fermions in small volumes, where the epsilon-expansion of chiral perturbation theory applies. Our data for the low lying Dirac eigenvalues, as well as mesonic correlation functions, are in agreement with analytical predictions. This allows us to extract values for the leading Low Energy Constants F_{pi} and Sigma.

  15. Simulating chiral quarks in the epsilon-regime of QCD

    E-print Network

    W. Bietenholz; T. Chiarappa; K. Jansen; K. -I. Nagai; S. Shcheredin

    2005-01-18

    We present simulation results for lattice QCD with chiral fermions in small volumes, where the epsilon-expansion of chiral perturbation theory applies. Our data for the low lying Dirac eigenvalues, as well as mesonic correlation functions, are in agreement with analytical predictions. This allows us to extract values for the leading Low Energy Constants F_{pi} and Sigma.

  16. Observations of Epsilon Lyrae by the Video Drift Method

    NASA Astrophysics Data System (ADS)

    Wasson, Rick; Nelson, Nancy; Nelson, Eric; Buehlman, William; Wilson, Earl; Zapata, Deanna

    2015-01-01

    The major components of the famous "double-double" star Epsilon Lyrae, STF2382AB and STF2383CD, were measured by the Video Team at the Apple Valley Double Star Workshop in 2013, using the Video Drift Method. The results are in reasonable agreement with other recent measures and predictions of the latest orbital solutions.

  17. Distribution of clostridial cry-like genes among Bacillus thuringiensis and Clostridium strains.

    PubMed

    Barloy, F; Lecadet, M M; Delécluse, A

    1998-04-01

    The presence of two cry-like genes first identified in Clostridium bifermentans subsp. malaysia CH18 was investigated in Clostridium species including 12 subspecies of Clostridium bifermentans, 13 strains of other members of Clostridia genus, and 13 different subspecies of Bacillus thuringiensis. Oligonucleotides designed to amplify the two toxin genes, cmb71 and cmb72, were used. We found that these genes are present in 80% of the Clostridium bifermentans strains tested and in 8% of the other Clostridium and Bacillus thuringiensis strains. PMID:9504991

  18. Constraints on the mass and mixing of the fourth generation quark from direct CP violation {epsilon}{prime}/{epsilon} and rare K decays

    SciTech Connect

    Huang, Chao-Shang; Huo, Wu-Jun; Wu, Yue-Liang

    2001-07-01

    We investigate the {epsilon}{prime}/{epsilon} for K{r_arrow}{pi}{pi} in a sequential fourth generation model. By giving the basic formulas for {epsilon}{prime}/{epsilon} in this model, we analyze the numerical results which are dependent on m{sub t{prime}} and the imaginary part of the fourth CKM factor, ImV{sub t{prime}s}{sup *}V{sub t{prime}d} (or V{sub t{prime}s}{sup *}V{sub t{prime}d} and the fourth generation CKM matrix phase {theta}). We find that, unlike the SM, when taking the central values of all parameters for {epsilon}{prime}/{epsilon}, the values of {epsilon}{prime}/{epsilon} can easily fit to the current experimental data for all values of hadronic matrix elements estimated from various approaches. Also, we show that the experimental values of {epsilon}{prime}/{epsilon} and rare K decays can provide a strong constraint on both mass and mixing of the fourth generation quark. When taking the values of hadronic matrix elements from the lattice or 1/N expansion calculations, a large region of the up-type quark mass m{sub t{prime}} is excluded.

  19. Clostridium beijerinckii and Clostridium difficile Detoxify Methylglyoxal by a Novel Mechanism Involving Glycerol Dehydrogenase

    PubMed Central

    Liyanage, Hemachandra; Kashket, Shelby; Young, Michael; Kashket, Eva R.

    2001-01-01

    In contrast to gram-negative bacteria, little is known about the mechanisms by which gram-positive bacteria degrade the toxic metabolic intermediate methylglyoxal (MG). Clostridium beijerinckii BR54, a Tn1545 insertion mutant of the NCIMB 8052 strain, formed cultures that contained significantly more (free) MG than wild-type cultures. Moreover, BR54 was more sensitive to growth inhibition by added MG than the wild type, suggesting that it has a reduced ability to degrade MG. The single copy of Tn1545 in this strain lies just downstream from gldA, encoding glycerol dehydrogenase. As a result of antisense RNA production, cell extracts of BR54 possess significantly less glycerol dehydrogenase activity than wild-type cell extracts (H. Liyanage, M. Young, and E. R. Kashket, J. Mol. Microbiol. Biotechnol. 2:87–93, 2000). Inactivation of gldA in both C. beijerinckii and Clostridium difficile gave rise to pinpoint colonies that could not be subcultured, indicating that glycerol dehydrogenase performs an essential function in both organisms. We propose that this role is detoxification of MG. To our knowledge, this is the first report of targeted gene disruption in the C. difficile chromosome. PMID:11319074

  20. Promoters and proteins from Clostridium thermocellum and uses thereof

    DOEpatents

    Wu, J. H. David; Newcomb, Michael

    2012-11-13

    The present invention relates to an inducible and a high expression nucleic acid promoter isolated from Clostridium thermocellum. These promoters are useful for directing expression of a protein or polypeptide encoded by a nucleic acid molecule operably associated with the nucleic acid promoters. The present invention also relates to nucleic acid constructs including the C. thermocellum promoters, and expression vectors and hosts containing such nucleic acid constructs. The present invention also relates to protein isolated from Clostridium thermocellum, including a repressor protein. The present invention also provides methods of using the isolated promoters and proteins from Clostridium thermocellum, including methods for directing inducible in vitro and in vivo expression of a protein or polypeptide in a host, and methods of producing ethanol from a cellulosic biomass.

  1. Botulinum neurotoxin homologs in non-Clostridium species.

    PubMed

    Mansfield, Michael J; Adams, Jeremy B; Doxey, Andrew C

    2015-01-30

    Clostridial neurotoxins (CNTs) are the deadliest toxins known and the causative agents of botulism and tetanus. Despite their structural and functional complexity, no CNT homologs are currently known outside Clostridium. Here, we report the first homologs of Clostridium CNTs within the genome of the rice fermentation organism Weissella oryzae SG25. One gene in W. oryzae S25 encodes a protein with a four-domain architecture and HExxH protease motif common to botulinum neurotoxins (BoNTs). An adjacent gene with partial similarity to CNTs is also present, and both genes seem to have been laterally transferred into the W. oryzae genome from an unknown source. Identification of mobile, CNT-related genes outside of Clostridium has implications for our understanding of the evolution of this important toxin family. PMID:25541486

  2. Metabolism of adenylylated nucleotides in Clostridium acetobutylicum.

    PubMed Central

    Balodimos, I A; Kashket, E R; Rapaport, E

    1988-01-01

    In response to the stresses imposed by temperature upshift or addition of butanol, Clostridium acetobutylicum cultures accumulated diadenosine-5',5'''-P1,P4-tetraphosphate (Ap4A) and adenosine 5'-P1,P4-tetraphospho-5'-guanosine (Ap4G) to high levels. The two adenylylated nucleotides were also accumulated in batch culture in the absence of imposed stresses when the clostridia switched from the acidogenic phase of growth to the solventogenic phase. Most of the adenylylated nucleotides were extracellular. The intracellular concentrations of these compounds were low throughout batch growth and in cells stressed by added butanol. In contrast to other procaryotes, these clostridia did not possess enzymes to degrade the dinucleotides, as shown with both intact cells and cell-free preparations. Our findings are consistent with the hypothesis that endogenously produced solvents are stressful to the cells, stimulating the synthesis of adenylylated nucleotides. The nucleotides accumulate extracellularly because they cannot be degraded and because the cell membranes are permeabilized by the solvents produced. PMID:3360745

  3. Anaerobic Degradation of Flavonoids by Clostridium orbiscindens

    PubMed Central

    Schoefer, Lilian; Mohan, Ruchika; Schwiertz, Andreas; Braune, Annett; Blaut, Michael

    2003-01-01

    An anaerobic, quercetin-degrading bacterium was isolated from human feces and identified as Clostridium orbiscindens by comparative 16S rRNA gene sequence analysis. The organism was tested for its ability to transform several flavonoids. The isolated C. orbiscindens strain converted quercetin and taxifolin to 3,4-dihydroxyphenylacetic acid; luteolin and eriodictyol to 3-(3,4-dihydroxyphenyl)propionic acid; and apigenin, naringenin, and phloretin to 3-(4-hydroxyphenyl)propionic acid, respectively. Genistein and daidzein were not utilized. The glycosidic bonds of luteolin-3-glucoside, luteolin-5-glucoside, naringenin-7-neohesperidoside (naringin), quercetin-3-glucoside, quercetin-3-rutinoside (rutin), and phloretin-2?-glucoside were not cleaved. Based on the intermediates and products detected, pathways for the degradation of the flavonol quercetin and the flavones apigenin and luteolin are proposed. To investigate the numerical importance of C. orbiscindens in the human intestinal tract, a species-specific oligonucleotide probe was designed and tested for its specificity. Application of the probe to fecal samples from 10 human subjects proved the presence of C. orbiscindens in 8 out of the 10 samples tested. The numbers ranged from 1.87 × 108 to 2.50 × 109 cells g of fecal dry mass?1, corresponding to a mean count of 4.40 × 108 cells g of dry feces?1. PMID:14532034

  4. Clostridium difficile Is an Autotrophic Bacterial Pathogen

    PubMed Central

    Köpke, Michael; Straub, Melanie; Dürre, Peter

    2013-01-01

    During the last decade, Clostridium difficile infection showed a dramatic increase in incidence and virulence in the Northern hemisphere. This incessantly challenging disease is the leading cause of antibiotic-associated and nosocomial infectious diarrhea and became life-threatening especially among elderly people. It is generally assumed that all human bacterial pathogens are heterotrophic organisms, being either saccharolytic or proteolytic. So far, this has not been questioned as colonization of the human gut gives access to an environment, rich in organic nutrients. Here, we present data that C. difficile (both clinical and rumen isolates) is also able to grow on CO2+H2 as sole carbon and energy source, thus representing the first identified autotrophic bacterial pathogen. Comparison of several different strains revealed high conservation of genes for autotrophic growth and showed that the ability to use gas mixtures for growth decreases or is lost upon prolonged culturing under heterotrophic conditions. The metabolic flexibility of C. difficile (heterotrophic growth on various substrates as well as autotrophy) could allow the organism in the gut to avoid competition by niche differentiation and contribute to its survival when stressed or in unfavorable conditions that cause death to other bacteria. This may be an important trait for the pathogenicity of C. difficile. PMID:23626782

  5. Clostridium botulinum in cattle and dairy products.

    PubMed

    Lindström, Miia; Myllykoski, Jan; Sivelä, Seppo; Korkeala, Hannu

    2010-04-01

    The use of plastic-wrapped and nonacidified silage as cattle feed has led to an increasing number of botulism outbreaks due to Clostridium botulinum Groups I-III in dairy cattle. The involvement of Groups I and II organisms in cattle botulism has raised concern of human botulism risk associated with the consumption of dairy products. Multiplication of C. botulinum in silage and in the gastrointestinal tract of cattle with botulism has been reported, thus contamination of the farm environment and raw milk, and further transmission through the dairy chain, are possible. The standard milk pasteurization treatment does not eliminate spores, and the intrinsic factors of many dairy products allow botulinal growth and toxin production. Although rare, several large botulism outbreaks due to both commercial and home-prepared dairy products have been reported. Factors explaining these outbreaks include most importantly temperature abuse, but also unsafe formulation, inadequate fermentation, insufficient thermal processing, post-process contamination, and lack of adequate quality control for adjunct ingredients were involved. The small number of outbreaks is probably explained by a low incidence of spores in milk, the presence of competitive bacteria in pasteurized milk and other dairy products, and growth-inhibitory combinations of intrinsic and extrinsic factors in cultured and processed dairy products. PMID:20301016

  6. Clostridium difficile: its disease and toxins.

    PubMed Central

    Lyerly, D M; Krivan, H C; Wilkins, T D

    1988-01-01

    Clostridium difficile is the etiologic agent of pseudomembranous colitis, a severe, sometimes fatal disease that occurs in adults undergoing antimicrobial therapy. The disease, ironically, has been most effectively treated with antibiotics, although some of the newer methods of treatment such as the replacement of the bowel flora may prove more beneficial for patients who continue to relapse with pseudomembranous colitis. The organism produces two potent exotoxins designated toxin A and toxin B. Toxin A is an enterotoxin believed to be responsible for the diarrhea and mucosal tissue damage which occur during the disease. Toxin B is an extremely potent cytotoxin, but its role in the disease has not been as well studied. There appears to be a cascade of events which result in the expression of the activity of these toxins, and these events, ranging from the recognition of a trisaccharide receptor by toxin A to the synergistic action of the toxins and their possible dissemination in the body, are discussed in this review. The advantages and disadvantages of the various assays, including tissue culture assay, enzyme immunoassay, and latex agglutination, currently used in the clinical diagnosis of the disease also are discussed. PMID:3144429

  7. Action of nitroheterocyclic drugs against Clostridium difficile.

    PubMed

    Kumar, Manish; Adhikari, Sudip; Hurdle, Julian G

    2014-10-01

    The nitroheterocyclic classes of drugs have a long history of use in treating anaerobic infections, as exemplified by metronidazole as a first-line treatment for mild-to-moderate Clostridium difficile infection (CDI). Since direct comparisons of the three major classes of nitroheterocyclic drugs (i.e. nitroimidazole, nitazoxanide and nitrofurans) and nitrosating agents against C. difficile are under-examined, in this study their actions against C. difficile were compared. Results show that whilst transient resistance occurs to metronidazole and nitazoxanide, stable resistance arises to nitrofurans upon serial passage. All compounds killed C. difficile at high concentrations in addition to the host defence nitrosating agent S-nitrosoglutathione (GSNO). This suggests that GSNO killing of C. difficile contributes to its efficacy in murine CDI. Although nitric oxide production could not be detected for the nitroheterocyclic drugs, the cellular response to metronidazole and nitrofurans has some overlap with the response to GSNO, causing significant upregulation of the hybrid-cluster protein Hcp that responds to nitrosative stress. These findings provide new insights into the action of nitroheterocyclic drugs against C. difficile. PMID:25129314

  8. The Changing Epidemiology of Clostridium difficile Infections

    PubMed Central

    Freeman, J.; Bauer, M. P.; Baines, S. D.; Corver, J.; Fawley, W. N.; Goorhuis, B.; Kuijper, E. J.; Wilcox, M. H.

    2010-01-01

    Summary: The epidemiology of Clostridium difficile infection (CDI) has changed dramatically during this millennium. Infection rates have increased markedly in most countries with detailed surveillance data. There have been clear changes in the clinical presentation, response to treatment, and outcome of CDI. These changes have been driven to a major degree by the emergence and epidemic spread of a novel strain, known as PCR ribotype 027 (sometimes referred to as BI/NAP1/027). We review the evidence for the changing epidemiology, clinical virulence and outcome of treatment of CDI, and the similarities and differences between data from various countries and continents. Community-acquired CDI has also emerged, although the evidence for this as a distinct new entity is less clear. There are new data on the etiology of and potential risk factors for CDI; controversial issues include specific antimicrobial agents, gastric acid suppressants, potential animal and food sources of C. difficile, and the effect of the use of alcohol-based hand hygiene agents. PMID:20610822

  9. Comparing the identification of Clostridium spp. by two Matrix-Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) mass spectrometry platforms to 16S rRNA PCR sequencing as a reference standard: a detailed analysis of age of culture and sample preparation.

    PubMed

    Chean, Roy; Kotsanas, Despina; Francis, Michelle J; Palombo, Enzo A; Jadhav, Snehal R; Awad, Milena M; Lyras, Dena; Korman, Tony M; Jenkin, Grant A

    2014-12-01

    We compared the identification of Clostridium species using mass spectrometry by two different Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) platforms (Bruker MS and Vitek MS) against 16S rRNA sequencing as the reference standard. We then examined the impact of different sample preparations and (on one of those platforms) age of bacterial colonial growth on the performance of the MALDI-TOF MS systems. We identified 10 different species amongst the 52 isolates by 16S rRNA sequencing, with Clostridium perfringens the most prevalent (n=30). Spectrometric analysis using Vitek MS correctly speciated 47/52 (90.4%) isolates and was not affected by the sample preparation used. Performance of the Bruker MS was dependent on sample preparation with correct speciation obtained for 36 of 52 (69.2%) isolates tested using the Direct Transfer [DT] protocol, but all 52 (100%) isolates were correctly speciated using either an Extended Direct Transfer [EDT] or a Full Formic Extraction [EX] protocol. We then examined the effect of bacterial colonial growth age on the performance of Bruker MS and found substantial agreement in speciation using DT (Kappa=0.62, 95% CI: 0.46-0.75), almost perfect agreement for EDT (Kappa=0.94, 95% CI: 0.86-1.00) and exact agreement for EX (Kappa=1.00) between different days. PMID:25230331

  10. Improved form of the low Reynolds number k-epsilon turbulence model

    Microsoft Academic Search

    G. H. Hoffman

    1974-01-01

    The behavior of the k-epsilon turbulence model at a turbulent-nonturbulent interface is examined to determine the turbulent diffusion coefficients sigma-k and sigma-epsilon. The analysis leads to a dilemma caused by the modeling of the generation term in the k and epsilon equations. Based on limiting values of sigma-k and sigma-epsilon derived from this analysis, a revised form of the low-Reynolds-number

  11. Thermolabile triose phosphate isomerase in a psychrophilic Clostridium.

    NASA Technical Reports Server (NTRS)

    Shing, Y. W.; Akagi, J. M.; Himes, R. H.

    1972-01-01

    It was found that a psychrophilic Clostridium contains a triose phosphate isomerase which is very labile at moderate temperatures. An investigation showed that the optimal growth temperature of the psychrophile was between 15 and 20 deg C. No growth occurred at 25 deg C. The thermostability of the glycolytic enzymes in the cell-free extracts of Clostridium sp. strain 69 was studied. The data obtained show that the triose phosphate isomerase is quite labile at moderate temperatures. The instability of the enzyme is sufficient to explain the low maximum growth temperature of the psychrophile.

  12. Numerical simulation of turbulent flow using the exponential expression of k-Epsilon equation

    Microsoft Academic Search

    Munehiko Hinatsu; Yoshiaki Kodama

    1987-01-01

    A numerical simulation of a turbulent flow around a wing section by use of the kappa-epsilon turbulence model is presented. In order to assure the positiveness of kappa and epsilon, the exponential expression of kappa-epsilon is used. The IAF method is adopted as a computational scheme. A treatment for the boundary condition on the wing surface is devised. A turbulent

  13. Computation of turbulent flows using an extended k-epsilon turbulence closure model

    Microsoft Academic Search

    Y.-S. Chen; S.-W. Kim

    1987-01-01

    An extended kappa-epsilon turbulence model is proposed and tested with successful results. An improved transport equation for the rate of dissipation of the turbulent kinetic energy, epsilon, is proposed. The proposed model gives more effective response to the energy production rate than does the standard kappa-epsilon turbulence model. An extra time scale of the production range is included in the

  14. Doppler Imaging of the Ap star epsilon Ursae Majoris: Ca, Cr, Fe, Mg, Mn, Ti, Sr

    Microsoft Academic Search

    T. Lueftinger; R. Kuschnig; N. E. Piskunov; W. W. Weiss

    2003-01-01

    The surface abundance structure of seven elements of the brightest chemically peculiar A-type star, epsilon Ursae Majoris (epsilon UMa, HD 112185, HR 4905) has been determined. Cr, Fe and, Mn are enhanced at the magnetic polar region of epsilon UMa while they avoid the magnetic equator. Sr seems to behave like Cr, Fe, and Mn, but is concentrated only at

  15. Determination of Re(. var epsilon. prime /. var epsilon. ) by the simultaneous detection of the four K sub L , S r arrow. pi. pi. decay modes

    SciTech Connect

    Patterson, J.R.; Barker, A.; Briere, R.A.; Gibbons, L.K.; Makoff, G.; Papadimitriou, V.; Somalwar, S.; Wah, Y.W.; Winstein, B.; Winston, R.; Woods, M.; Yamamoto, H. (The Enrico Fermi Institute, The University of Chicago, Chicago, IL (USA) The Department of Physics, The University of Chicago, Chicago, Illinois 60637 (USA)); Swallow, E. (Department of Physics, Elmhurst College, Elmhurst, Illinois 60126 (USA) Department of Physics, The University of Chicago, Chicago, Illinois 60637 (USA)); Bock, G.J.; Coleman, R.; Enagonio, J.; Hsiung, Y.B.; Stanfield, K.; Stefanski, R.; Yamanaka, T. (Fermi National Accelerator Laboratory, Batavia, Illinois 60510 (USA)); Blair, G.; Gollin, G.D.; Karlsson, M.; Okamitsu, J.K.; Tschirhart, R. (Department of Physics, Princeton University, Princeton, New Jersey 08544 (USA)); Brisson, J.C.; Debu, P.; Peyaud, B.; Turlay, R.; Vallage, B. (Department de Physique des Particules Elementaires, Centre d'Etudes Nucleaires de Saclay, F-91191 Gif-sur-Yvette CEDEX,

    1990-03-26

    The E731 experiment at Fermilab has searched for direct {ital CP} violation in {ital K}{sup 0}{r arrow}{pi}{pi}, which is parametrized by {var epsilon}{prime}/{var epsilon}. For the first time all four of the {ital K}{sub {ital L},}{ital S}{r arrow}{pi}{pi} modes were collected simultaneously, which greatly facilitated studies of systematic uncertainty. We find Re({var epsilon}{prime}/{var epsilon})={minus}0.0004{plus minus}0.0014(stat){plus minus}0.0006(syst). The result provides no evidence for direct {ital CP} violation.

  16. Determination of Re(. var epsilon. prime \\/. var epsilon. ) by the simultaneous detection of the four K sub L , S r arrow. pi. pi. decay modes

    Microsoft Academic Search

    J. R. Patterson; A. Barker; R. A. Briere; L. K. Gibbons; G. Makoff; V. Papadimitriou; S. Somalwar; Y. W. Wah; B. Winstein; R. Winston; M. Woods; H. Yamamoto; E. Swallow; G. J. Bock; R. Coleman; J. Enagonio; Y. B. Hsiung; K. Stanfield; R. Stefanski; T. Yamanaka; G. Blair; G. D. Gollin; M. Karlsson; J. K. Okamitsu; R. Tschirhart; J. C. Brisson; P. Debu; B. Peyaud; R. Turlay; B. Vallage

    1990-01-01

    The E731 experiment at Fermilab has searched for direct {ital CP} violation in {ital K}°{r arrow}ÏÏ, which is parametrized by {var epsilon}â²\\/{var epsilon}. For the first time all four of the {ital K}{sub {ital L},}{ital S}{r arrow}ÏÏ modes were collected simultaneously, which greatly facilitated studies of systematic uncertainty. We find Re({var epsilon}â²\\/{var epsilon})=-0.0004{plus minus}0.0014(stat){plus minus}0.0006(syst). The result provides no evidence

  17. Form and Function of Clostridium thermocellum Biofilms

    PubMed Central

    Dumitrache, Alexandru; Allen, Grant; Liss, Steven N.; Lynd, Lee R.

    2013-01-01

    The importance of bacterial adherence has been acknowledged in microbial lignocellulose conversion studies; however, few reports have described the function and structure of biofilms supported by cellulosic substrates. We investigated the organization, dynamic formation, and carbon flow associated with biofilms of the obligately anaerobic cellulolytic bacterium Clostridium thermocellum 27405. Using noninvasive, in situ fluorescence imaging, we showed biofilms capable of near complete substrate conversion with a characteristic monolayered cell structure without an extracellular polymeric matrix typically seen in biofilms. Cell division at the interface and terminal endospores appeared throughout all stages of biofilm growth. Using continuous-flow reactors with a rate of dilution (2 h?1) 12-fold higher than the bacterium's maximum growth rate, we compared biofilm activity under low (44 g/liter) and high (202 g/liter) initial cellulose loading. The average hydrolysis rate was over 3-fold higher in the latter case, while the proportions of oligomeric cellulose hydrolysis products lost from the biofilm were 13.7% and 29.1% of the total substrate carbon hydrolyzed, respectively. Fermentative catabolism was comparable between the two cellulose loadings, with ca. 4% of metabolized sugar carbon being utilized for cell production, while 75.4% and 66.7% of the two cellulose loadings, respectively, were converted to primary carbon metabolites (ethanol, acetic acid, lactic acid, carbon dioxide). However, there was a notable difference in the ethanol-to-acetic acid ratio (g/g), measured to be 0.91 for the low cellulose loading and 0.41 for the high cellulose loading. The results suggest that substrate availability for cell attachment rather than biofilm colonization rates govern the efficiency of cellulose conversion. PMID:23087042

  18. A k-epsilon modeling of near wall turbulence

    NASA Technical Reports Server (NTRS)

    Yang, Z.; Shih, T. H.

    1991-01-01

    A k-epsilon model is proposed for turbulent bounded flows. In this model, the turbulent velocity scale and turbulent time scale are used to define the eddy viscosity. The time scale is shown to be bounded from below by the Kolmogorov time scale. The dissipation equation is reformulated using the time scale, removing the need to introduce the pseudo-dissipation. A damping function is chosen such that the shear stress satisfies the near wall asymptotic behavior. The model constants used are the same as the model constants in the commonly used high turbulent Reynolds number k-epsilon model. Fully developed turbulent channel flows and turbulent boundary layer flows over a flat plate at various Reynolds numbers are used to validate the model. The model predictions were found to be in good agreement with the direct numerical simulation data.

  19. EISCAT observations during MAC/SINE and MAC/Epsilon

    NASA Technical Reports Server (NTRS)

    Roettger, J.; Hoppe, U.-P.; Hall, C.

    1989-01-01

    The EISCAT incoherent scatter radar facility in Tromsoe, Norway was operated during the MAC/SINE campaign for 78 hours in the period 10 June to 17 July 1987, and during the MAC/Epsilon campaign for 90 hours in the period 15 October to 5 November 1987. The VHF (224 MHz) radar operations during MAC/SINE yielded most interesting observations of strong coherent echoes from the mesopause region. Characteristic data of these polar mesospheric summer echoes are presented. The UHF (933 MHz) radar operations during MAC/Epsilon were done with 18 deg off zenith beam and allows the deduction of meridonal and horizontal wind components as well as radial velocity spectra in addition to the usual electron density profiles in the D and lower E regions. Some results from the VHF and UHF radars indicating the presence of gravity waves are examined.

  20. The Epsilon Aurigae secondary - A binary embedded within a disk?

    NASA Technical Reports Server (NTRS)

    Lissauer, J. J.; Backman, D. E.

    1984-01-01

    The eclipse characteristics of Epsilon Aurigae, which is an F supergiant in a spectroscopic binary system, imply a secondary that is elongated, measuring about 10 AU by less than 1 AU, with a mass of about 16 solar masses. Recent IR observations, however, indicate a color temperature of about 500 K and a luminosity that is apparently less than 1 percent that of a 16-solar mass star. It is presently suggested that the secondary consists of a close binary embedded in an optically thick disk, which is viewed edge-on. The observed low luminosity of the secondary would then be due to two 8-solar mass stars' luminosity, which is about 10 percent that of a 16-solar mass star, together with the loss of 90 percent of the emitted energy through escape from the poles of the disk. This hypothesis is discussed in relation to radio and UV observations of Epsilon Aurigae.

  1. An obvious isospin breaking correction to epsilon' of kinematical origin

    E-print Network

    Mahiko Suzuki

    2001-02-08

    Isospin breaking correction to the $\\Delta I = {1/2}$ decay of $K\\to\\pi\\pi$ generates a large enough contribution to $\\epsilon'$ through an induced $\\Delta I = {3/2}$ amplitude in $K_L$ decay. Aside from the $\\pi$-$\\eta$ and $\\pi$-$\\eta'$ mixing contributions, there is a correction of kinematical origin due to the final-state $\\pi^{\\pm}$-$\\pi^0$ mass difference, which is unambiguously calculable from the low-energy off-shell behavior of the $K\\to\\pi\\pi$ amplitude. This correction to $\\epsilon'$ reduces the isospin breaking parameter $\\Omega_{IB}$ by 0.06, which is nearly one half of the $\\pi$-$\\eta$ mixing effect computed with chiral Lagrangians to $O(p^2)$.

  2. Production of 1,3-Propanediol from Glycerol by Clostridium acetobutylicum and Other Clostridium Species

    PubMed Central

    Forsberg, Cecil W.

    1987-01-01

    Glycerol was fermented with the production of 1,3-propanediol as the major fermentation product by four strains of Clostridium acetobutylicum, six of C. butylicum, two of C. beijerinckii, one of C. kainantoi, and three of C. butyricum. 1,3-Propanediol was identified by its retention times in gas chromatography and high-pressure liquid chromatography and by its mass spectrum. During growth of C. butylicum B593 in a chemostat culture at pH 6.5, 61% of the glycerol fermented was converted to 1,3-propanediol. When the pH was decreased to 4.9, growth and 1,3-propanediol production were substantially reduced. PMID:16347311

  3. Magnetoelastic nature of solid oxygen epsilon-phase structure

    Microsoft Academic Search

    Helen V. Gomonay; Vadim M. Loktev

    2007-01-01

    For a long time a crystal structure of high-pressure epsilon-phase of solid\\u000aoxygen was a mistery. Basing on the results of recent experiments that have\\u000asolved this riddle we show that the magnetic and crystal structure of\\u000aepsilon-phase can be explained by strong exchange interactions of\\u000aantiferromagnetic nature. The singlet state implemented on quaters of O2\\u000amolecules has the minimal

  4. An improved k-epsilon model for near wall turbulence

    Microsoft Academic Search

    T. H. Shih; Andrew T. Hsu

    1991-01-01

    An improved k-epsilon model for low Reynolds number turbulence near a wall is presented. In the first part of this work, the near-wall asymptotic behavior of the eddy viscosity and the pressure transport term in the turbulent kinetic energy equation are analyzed. Based on these analyses, a modified eddy viscosity model with the correct near-wall behavior is suggested, and a

  5. Advanced k-epsilon modeling of heat transfer

    Microsoft Academic Search

    Okey Kwon; Forrest E. Ames

    1995-01-01

    This report describes two approaches to low Reynolds-number k-epsilon turbulence modeling which formulate the eddy viscosity on the wall-normal component of turbulence and a length scale. The wall-normal component of turbulence is computed via integration of the energy spectrum based on the local dissipation rate and is bounded by the isotropic condition. The models account for the anisotropy of the

  6. Simulations in the epsilon-Regime of Chiral Perturbation Theory

    E-print Network

    K. -I. Nagai; W. Bietenholz; T. Chiarappa; K. Jansen; S. Shcheredin

    2003-09-12

    We discuss the potential of Ginsparg-Wilson fermion simulations in the epsilon-regime of chiral perturbation theory, regarding the determination of the leading low energy constants of the effective chiral Lagrangian. It turns out to be very hard to measure observables in the topologically trivial sector. There a huge statistics would be required, due to the frequent occurrence of very small eigenvalues. Moreover, contact with chiral perturbation theory is only established if the physical volume of the system is sufficiently large.

  7. A kappa-epsilon calculation of transitional boundary layers

    NASA Technical Reports Server (NTRS)

    Yang, Z.; Shih, T. H.

    1992-01-01

    A recently proposed kappa-epsilon model for low Reynolds number turbulent flows was modified by introducing a new damping function f(sub mu). The modified model is used to calculate the transitional boundary layer over a flat plate with different freestream turbulence levels. It is found that the model could mimic the transitional flow. However, the predicted transition is found to be sensitive to the initial conditions.

  8. Clostridium difficile in retail meat and processing plants in Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The incidence and severity of disease associated with toxigenic Clostridium difficile (Cd) have increased in hospitals in North America from the emergence of newer, more virulent strains of Cd. Toxigenic Cd has been isolated from food animals and retail meat with potential implications of transfer ...

  9. 3-Methylindole production is regulated in Clostridium scatologenes ATCC 25775

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aims: 3-Methylindole (3-MI) is a degradation product of L-tryptophan and is both an animal waste malodorant and threat to ruminant health. Culture conditions which influence 3-MI production in Clostridium scatologenes ATCC 25775 were investigated. Methods and Results: Cells cultured in anaerobic ...

  10. Recommendations for Surveillance of Clostridium difficile –Associated Disease

    Microsoft Academic Search

    L. Clifford McDonald; Bruno Coignard; Erik Dubberke; Xiaoyan Song; Teresa Horan; Preeta K. Kutty

    2007-01-01

    background. The epidemiology of Clostridium difficile-associated disease (CDAD) is changing, with evidence of increased incidence and severity. However, the understanding of the magnitude of and reasons for this change is currently hampered by the lack of standardized surveillance methods. objective and methods. An ad hoc C. difficile surveillance working group was formed to develop interim surveillance definitions and recommendations based

  11. Preventing downstream Clostridium difficile infections with upstream antibiotic management

    Microsoft Academic Search

    Alpin D. Malkan; Stephen J. Scholand

    Clostridium difficile infection (CDI) remains a devastating cause of hospital-acquired diarrhea. Treatment modalities have centered traditionally on two antibiotics, metronidazole and oral vancomycin. Both drugs, however, have been associated with variable relapse rates up to 20%. Fidaxomicin, a new oral agent with targeted C. difficile activity, may reduce the chance of relapse, but has not yet entered mainstream clinical practice.

  12. Varied prevalence of Clostridium difficile in an integrated swine operation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to compare the prevalence of Clostridium difficile among different age and production groups of swine in a vertically integrated swine operation in Texas in 2006 and to compare our isolates to other animal and human isolates. Preliminary results are based on 131 C. d...

  13. Rifampin and Rifaximin Resistance in Clinical Isolates of Clostridium difficile

    Microsoft Academic Search

    Jennifer R. O'Connor; Minerva A. Galang; Susan P. Sambol; David W. Hecht; Gayatri Vedantam; Dale N. Gerding; Stuart Johnson

    2008-01-01

    Rifaximin, a poorly absorbed rifamycin derivative, is a promising alternative for the treatment of Clostridium difficile infections. Resistance to this agent has been reported, but no commercial test for rifaximin resistance exists and the molecular basis of this resistance has not been previously studied in C. difficile. To evaluate whether the rifampin Etest would be a suitable substitute for rifaximin

  14. Cryptic Polyketide Synthase Genes in Non-Pathogenic Clostridium SPP

    Microsoft Academic Search

    Swantje Behnken; Christian Hertweck

    2012-01-01

    Modular type I polyketide synthases (PKS) produce a vast array of bacterial metabolites with highly diverse biological functions. Notably, all known polyketides were isolated from aerobic bacteria, and yet no example has been reported for strict anaerobes. In this study we explored the diversity and distribution of PKS genes in the genus Clostridium. In addition to comparative genomic analyses combined

  15. Possible Seasonality of Clostridium difficile in Retail Meat, Canada

    PubMed Central

    Reid-Smith, Richard J.; Staempfli, Henry R.; Daignault, Danielle; Janecko, Nicol; Avery, Brent P.; Martin, Hayley; Thomspon, Angela D.; McDonald, L. Clifford; Limbago, Brandi; Weese, J. Scott

    2009-01-01

    We previously reported Clostridium difficile in 20% of retail meat in Canada, which raised concerns about potential foodborne transmissibility. Here, we studied the genetic diversity of C. difficile in retail meats, using a broad Canadian sampling infrastructure and 3 culture methods. We found 6.1% prevalence and indications of possible seasonality (highest prevalence in winter). PMID:19402975

  16. ID Learning Unit: Understanding and Interpreting Testing for Clostridium difficile

    PubMed Central

    Solomon, Daniel A.; Milner, Danny A.

    2014-01-01

    Understanding and interpreting the molecular tests for Clostridium difficile is challenging because there are several different types of assays and most laboratories combine multiple tests in order to assess for presence of disease. This learning unit demonstrates the basic principles of each test along with its strengths and weaknesses, and illustrates how the tests are used in clinical practice. PMID:25734081

  17. Clostridium difficile prevalence in an integrated swine operation in Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recently there has been an epidemic of human disease in North America caused by the bacterium Clostridium difficile (Cd). It appears to be a new strain that is more virulent than previous strains, produces more toxins, and causes more severe disease (McDonald et al., 2005). The origin of the new s...

  18. Clostridium difficile in mixed populations of animals and humans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objectives: Since 2003, there has been an emergence of BI/NAP1 strain of Clostridium difficile (Cd) in North American hospitals. The origins of this epidemic strain have yet to be determined. However, PFGE analysis has shown ~80% similarity between this strain and some swine isolates. The objecti...

  19. Magnetic Activity Cycles in the Exoplanet Host Star epsilon Eridani

    NASA Astrophysics Data System (ADS)

    Metcalfe, T. S.; Buccino, A. P.; Brown, B. P.; Mathur, S.; Soderblom, D. R.; Henry, T. J.; Mauas, P. J. D.; Petrucci, R.; Hall, J. C.; Basu, S.

    2013-02-01

    The active K2 dwarf epsilon Eri has been extensively characterized both as a young solar analog and more recently as an exoplanet host star. As one of the nearest and brightest stars in the sky, it provides an unparalleled opportunity to constrain stellar dynamo theory beyond the Sun. We confirm and document the 3-year magnetic activity cycle in epsilon Eri originally reported by Hatzes and coworkers, and we examine the archival data from previous observations spanning 45 years. The data show coexisting 3-year and 13-year periods leading into a broad activity minimum that resembles a Maunder minimum-like state, followed by the resurgence of a coherent 3-year cycle. The nearly continuous activity record suggests the simultaneous operation of two stellar dynamos with cycle periods of 2.95 ± 0.03 years and 12.7 ± 0.3 years, which, by analogy with the solar case, suggests a revised identification of the dynamo mechanisms that are responsible for the so-called "active" and "inactive" sequences as proposed by Böhm-Vitense. Finally, based on the observed properties of epsilon Eri, we argue that the rotational history of the Sun is what makes it an outlier in the context of magnetic cycles observed in other stars (as also suggested by its Li depletion), and that a Jovian-mass companion cannot be the universal explanation for the solar peculiarities.

  20. Advanced k-epsilon modeling of heat transfer

    NASA Technical Reports Server (NTRS)

    Kwon, Okey; Ames, Forrest E.

    1995-01-01

    This report describes two approaches to low Reynolds-number k-epsilon turbulence modeling which formulate the eddy viscosity on the wall-normal component of turbulence and a length scale. The wall-normal component of turbulence is computed via integration of the energy spectrum based on the local dissipation rate and is bounded by the isotropic condition. The models account for the anisotropy of the dissipation and the reduced mixing length due to the high strain rates present in the near-wall region. The turbulent kinetic energy and its dissipation rate were computed from the k and epsilon transport equations of Durbin. The models were tested for a wide range of turbulent flows and proved to be superior to other k-epsilon models, especially for nonequilibrium anisotropic flows. For the prediction of airfoil heat transfer, the models included a set of empirical correlations for predicting laminar-turbulent transition and laminar heat transfer augmentation due to the presence of freestream turbulence. The predictions of surface heat transfer were generally satisfactory.

  1. Molecular Characterization of the Clusters of Genes Encoding the Botulinum Neurotoxin Complex in Clostridium botulinum ( Clostridium argentinense ) Type G and Nonproteolytic Clostridium botulinum Type B

    Microsoft Academic Search

    Manju Bhandari; Kathryn D. Campbell; Matthew D. Collins; Alison K. East

    1997-01-01

    .   The cluster of genes encoding components of the progenitor botulinum neurotoxin complex has been mapped and cloned in Clostridium botulinum type G strain ATCC 27322. Determination of the nucleotide sequence of the region has revealed open reading frames encoding\\u000a nontoxic components of the complex, upstream of the gene encoding BoNT\\/G (botG). The arrangement of these genes differs from that

  2. Development of a triplex real-time PCR assay for the simultaneous detection of Clostridium beijerinckii, Clostridium sporogenes and Clostridium tyrobutyricum in milk.

    PubMed

    Morandi, Stefano; Cremonesi, Paola; Silvetti, Tiziana; Castiglioni, Bianca; Brasca, Milena

    2015-08-01

    Clostridium beijerinckii, Clostridium sporogenes and Clostridium tyrobutyricum are considered the leading bacteria implicated in late blowing defects affecting semi-hard and hard cheese production. The aim of this study was to develop a multiplex Real-Time PCR (qPCR) analysis for a rapid and simultaneous detection of C. beijerinckii, C. sporogenes and C. tyrobutyricum, using specific primers respectively targeting the nifH, gerAA and enr genes. The limits of detection in raw milk were 300 CFU/50 mL in the case of C. beijerinckii, 2 CFU/50 mL for C. sporogenes and 5 CFU/50 mL for C. tyrobutyricum spores. The qPCR method was applied to artificially contaminated raw milk samples, and molecular quantification showed good correlation (R(2) = 0.978) with microbiological counting. Our results demonstrate that this method, combined with a DNA extraction protocol optimized for spore lysis, could be a useful tool for the direct quantification of the considered clostridia species. PMID:25870135

  3. Development and validation of a multiplex real-time PCR for detection of Clostridium chauvoei and Clostridium septicum.

    PubMed

    Lange, Martin; Neubauer, Heinrich; Seyboldt, Christian

    2010-08-01

    Clostridium chauvoei is the causative agent of blackleg in cattle and sheep. The clinical symptoms of this severe disease are very similar to that of malignant edema (Clostridium septicum), infections of other Clostridium species belonging to the gas edema complex, and anthrax (Bacillus anthracis). C. chauvoei and C. septicum are closely related taxa and share many phenotypic properties hampering diagnosis by using traditional microbiological methods. Thus, there is a need for a fast and reliable identification method for specific detection of both species in clinical samples. The multiplex real-time PCR assay presented here is based on the detection of the spo0A gene and enables the simultaneous identification of C. chauvoei and C. septicum. The assay design includes an amplification control DNA template for the recognition of PCR-inhibitors. Assay validation was performed using a collection of 29 C. chauvoei, 38 C. septicum strains and 26 strains of other Clostridium species. Furthermore, the real-time PCR assay was successfully tested on tissue samples from 19 clinical blackleg cases. The assay allowed the reliable detection of one picogram DNA which represents approximate 239 genome equivalents. PMID:20362050

  4. Switchgrass (Panicum virgatum) fermentation by Clostridium thermocellum and Clostridium beijerinckii sequential culture: effect of feedstock particle size on gas production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fermentation of cellulosic biomass can be done in a single step with cellulolytic, solventogenic bacteria, such as Clostridium thermocellum. However, the suite of products is limited in consolidated bioprocessing. Fortunately, the thermophilic nature of C. thermocellum can be exploited in sequenti...

  5. Aerial Dissemination of Clostridium difficile spores

    PubMed Central

    Roberts, Katherine; Smith, Caroline F; Snelling, Anna M; Kerr, Kevin G; Banfield, Kathleen R; Sleigh, P Andrew; Beggs, Clive B

    2008-01-01

    Background Clostridium difficile-associated diarrhoea (CDAD) is a frequently occurring healthcare-associated infection, which is responsible for significant morbidity and mortality amongst elderly patients in healthcare facilities. Environmental contamination is known to play an important contributory role in the spread of CDAD and it is suspected that contamination might be occurring as a result of aerial dissemination of C. difficile spores. However previous studies have failed to isolate C. difficile from air in hospitals. In an attempt to clarify this issue we undertook a short controlled pilot study in an elderly care ward with the aim of culturing C. difficile from the air. Methods In a survey undertaken during February (two days) 2006 and March (two days) 2007, air samples were collected using a portable cyclone sampler and surface samples collected using contact plates in a UK hospital. Sampling took place in a six bedded elderly care bay (Study) during February 2006 and in March 2007 both the study bay and a four bedded orthopaedic bay (Control). Particulate material from the air was collected in Ringer's solution, alcohol shocked and plated out in triplicate onto Brazier's CCEY agar without egg yolk, but supplemented with 5 mg/L of lysozyme. After incubation, the identity of isolates was confirmed by standard techniques. Ribotyping and REP-PCR fingerprinting were used to further characterise isolates. Results On both days in February 2006, C. difficile was cultured from the air with 23 samples yielding the bacterium (mean counts 53 – 426 cfu/m3 of air). One representative isolate from each of these was characterized further. Of the 23 isolates, 22 were ribotype 001 and were indistinguishable on REP-PCR typing. C. difficile was not cultured from the air or surfaces of either hospital bay during the two days in March 2007. Conclusion This pilot study produced clear evidence of sporadic aerial dissemination of spores of a clone of C. difficile, a finding which may help to explain why CDAD is so persistent within hospitals and difficult to eradicate. Although preliminary, the findings reinforce concerns that current C. difficile control measures may be inadequate and suggest that improved ward ventilation may help to reduce the spread of CDAD in healthcare facilities. PMID:18218089

  6. Clostridium difficile associated infection, diarrhea and colitis

    PubMed Central

    Hookman, Perry; Barkin, Jamie S

    2009-01-01

    A new, hypervirulent strain of Clostridium difficile, called NAP1/BI/027, has been implicated in C. difficile outbreaks associated with increased morbidity and mortality since the early 2000s. The epidemic strain is resistant to fluoroquinolones in vitro, which was infrequent prior to 2001. The name of this strain reflects its characteristics, demonstrated by different typing methods: pulsed-field gel electrophoresis (NAP1), restriction endonuclease analysis (BI) and polymerase chain reaction (027). In 2004 and 2005, the US Centers for Disease Control and Prevention (CDC) emphasized that the risk of C. difficile-associated diarrhea (CDAD) is increased, not only by the usual factors, including antibiotic exposure, but also gastrointestinal surgery/manipulation, prolonged length of stay in a healthcare setting, serious underlying illness, immune-compromising conditions, and aging. Patients on proton pump inhibitors (PPIs) have an elevated risk, as do peripartum women and heart transplant recipients. Before 2002, toxic megacolon in C. difficile-associated colitis (CDAC), was rare, but its incidence has increased dramatically. Up to two-thirds of hospitalized patients may be infected with C. difficile. Asymptomatic carriers admitted to healthcare facilities can transmit the organism to other susceptible patients, thereby becoming vectors. Fulminant colitis is reported more frequently during outbreaks of C. difficile infection in patients with inflammatory bowel disease (IBD). C. difficile infection with IBD carries a higher mortality than without underlying IBD. This article reviews the latest information on C. difficile infection, including presentation, vulnerable hosts and choice of antibiotics, alternative therapies, and probiotics and immunotherapy. We review contact precautions for patients with known or suspected C. difficile-associated disease. Healthcare institutions require accurate and rapid diagnosis for early detection of possible outbreaks, to initiate specific therapy and implement effective control measures. A comprehensive C. difficile infection control management rapid response team (RRT) is recommended for each health care facility. A communication network between RRTs is recommended, in coordination with each country’s department of health. Our aim is to convey a comprehensive source of information and to guide healthcare professionals in the difficult decisions that they face when caring for these oftentimes very ill patients. PMID:19340897

  7. A complicated quasicrystal approximant epsilon16 predicted by the strong-reflections approach.

    PubMed

    Li, Mingrun; Sun, Junliang; Oleynikov, Peter; Hovmöller, Sven; Zou, Xiaodong; Grushko, Benjamin

    2010-02-01

    The structure of a complicated quasicrystal approximant epsilon(16) was predicted from a known and related quasicrystal approximant epsilon(6) by the strong-reflections approach. Electron-diffraction studies show that in reciprocal space, the positions of the strongest reflections and their intensity distributions are similar for both approximants. By applying the strong-reflections approach, the structure factors of epsilon(16) were deduced from those of the known epsilon(6) structure. Owing to the different space groups of the two structures, a shift of the phase origin had to be applied in order to obtain the phases of epsilon(16). An electron-density map of epsilon(16) was calculated by inverse Fourier transformation of the structure factors of the 256 strongest reflections. Similar to that of epsilon(6), the predicted structure of epsilon(16) contains eight layers in each unit cell, stacked along the b axis. Along the b axis, epsilon(16) is built by banana-shaped tiles and pentagonal tiles; this structure is confirmed by high-resolution transmission electron microscopy (HRTEM). The simulated precession electron-diffraction (PED) patterns from the structure model are in good agreement with the experimental ones. Epsilon(16) with 153 unique atoms in the unit cell is the most complicated approximant structure ever solved or predicted. PMID:20101079

  8. Discrimination of clostridium species using a magnetic bead based hybridization assay

    NASA Astrophysics Data System (ADS)

    Pahlow, Susanne; Seise, Barbara; Pollok, Sibyll; Seyboldt, Christian; Weber, Karina; Popp, Jürgen

    2014-05-01

    Clostridium chauvoei is the causative agent of blackleg, which is an endogenous bacterial infection. Mainly cattle and other ruminants are affected. The symptoms of blackleg are very similar to those of malignant edema, an infection caused by Clostridium septicum. [1, 2] Therefore a reliable differentiation of Clostridium chauvoei from other Clostridium species is required. Traditional microbiological detection methods are time consuming and laborious. Additionally, the unique identification is hindered by the overgrowing tendency of swarming Clostridium septicum colonies when both species are present. [1, 3, 4] Thus, there is a crucial need to improve and simplify the specific detection of Clostridium chauvoei and Clostridium septicum. Here we present an easy and fast Clostridium species discrimination method combining magnetic beads and fluorescence spectroscopy. Functionalized magnetic particles exhibit plentiful advantages, like their simple manipulation in combination with a large binding capacity of biomolecules. A specific region of the pathogenic DNA is amplified and labelled with biotin by polymerase chain reaction (PCR). These PCR products were then immobilized on magnetic beads exploiting the strong biotin-streptavidin interaction. The specific detection of different Clostridium species is achieved by using fluorescence dye labeled probe DNA for the hybridization with the immobilized PCR products. Finally, the samples were investigated by fluorescence spectroscopy. [5

  9. Electric field measurements during the MAC/EPSILON campaign

    NASA Technical Reports Server (NTRS)

    Croskey, C. L.; Hale, L. C.; Mitchell, J. D.; Schmidlin, F. J.; Hoppe, U.-P.

    1990-01-01

    The MAC/EPSILON observational campaign in northern Norway involved the taking of three-axis electric field measurements of the middle atmosphere by means of five rocketborne payloads during October and November, 1987. Simultaneous horizontal electric field measurements made by two of the rocket flights were in general agreement in their limited overlap region. The more extensive horizontal E-field measurements exhibited a decreasing mapping function with decreasing altitude, thereby indicating the presence of fields from a local auroral patch. Small-scale variations in the horizontal fields of the lights were similar to observed wavelike variations in the neutral wind field.

  10. Light focusing using epsilon-near-zero metamaterials

    SciTech Connect

    Zhu, Weiren, E-mail: weiren.zhu@monash.edu; Premaratne, Malin [Advanced Computing and Simulation Laboratory (A chi L), Department of Electrical and Computer Systems Engineering, Monash University, Clayton, Victoria 3800 (Australia)] [Advanced Computing and Simulation Laboratory (A chi L), Department of Electrical and Computer Systems Engineering, Monash University, Clayton, Victoria 3800 (Australia); Si, Li-Ming, E-mail: lms@bit.edu.cn [Beijing Key Laboratory of Millimeter Wave and Terahertz Technology, Department of Electronic Engineering, School of Information and Electronics, Beijing Institute of Technology, Beijing 100081 (China)] [Beijing Key Laboratory of Millimeter Wave and Terahertz Technology, Department of Electronic Engineering, School of Information and Electronics, Beijing Institute of Technology, Beijing 100081 (China)

    2013-11-15

    We present a strategy of focusing light using epsilon-near-zero metamaterials with embedded dielectric cylinder. The focusing mechanism is analytically investigated, and its accuracy is substantiated by rigorous full-wave simulations. It is found that the focusing intensity is highly depend on the embedded medium and its size, and the magnetic field amplitude of the focused beam itself can reach as high as 98.2 times the incident field. Owing to its versatility, the proposed light focusing system is sure to find applications in fields such as bio-sensing and in nonlinear optics.

  11. Epsilon Canis Majoris and the ionization of the local cloud

    NASA Technical Reports Server (NTRS)

    Vallerga, J. V.; Welsh, B. Y.

    1995-01-01

    The Lyman continuum radiation from the brightest extreme ultraviolet (EUV) source, the B2 II star epsilon Canis Majoris (Adara), is so intense that it dominates the local stellar EUV radiation field at wavelengths longer than 450 A and therefore sets a lower limit to the ionization of hydrogen in the Local Cloud. Using the EUV (70-730 A) spectrum of epsilon CMa taken with the Extreme Ultraviolet Explorer Satellite (EUVE) and simple models that extrapolate this spectrum to the Lyman edge at 912 A, we have determined the local interstellar hydrogen photionizatin parameter Gamma solely from epsilon CMa to be 1.1 x 10(exp -15)/s. This fiugre is a factor of 7 greater than previous estimates of Gamma calculated for all nearby stars combined (Bruhweiler & Cheng 1988). Using measured values of the density and temperature of neutral interstellar hydrogen gas in the Local Cloud, we derive a particle density of ionized hydrogen n(H(+)) and electrons n(sub e) of 0.015-0.019/cu cm assuming ionization equilibrium and a helium ionization fraction of less than 20%. These values correspond to a hydrogen ionizatin fraction, chi(sub H) from 19% to 15%, respectively. The range of these derived quantities is due to the uncertainties in the local values of the neutral hydrogen and helium interstellar densities derived from both (1) solar backscatter measurements of Ly alpha lines of hydrogen and helium (1216 and 584 A), and (2) the average neutral densities along the line of sight to nearby stars. The local proton density produced by epsilon CMa is enough to allow the ionization mechanism of Ripken & Fahr (1983) to work at the heliopause and explain the discrepancy between the neutral hydrogen density derived from solar backscatter measurements and line-of-sight averages to nearby stars. A large value of electron density in the Local Cloud of n(sub e) is approximately 0.3-0.7/cu cm (T = 7000 K) has recently been reported by Lallement et al. (1994) using observations of Mg II and Mg I toward Sirius A. We show that if such a high value exists, it cannot result from the EUV stellar radiation field and, therefore, must be due to a strong diffuse source of EUV radiation.

  12. The 1982-1984 Eclipse of Epsilon Aurigae

    NASA Technical Reports Server (NTRS)

    Stencel, R. E. (editor)

    1985-01-01

    A workshop proceedings concerned with the new data collected during the 1982-1984 eclipse period of the 27-year system Epsilon Aurigae is presented. This binary star has been a classic problem in astrophysics because the opaque eclipsing object is nonstellar, and probably disk shaped. Invited papers concerning the history of the system, optical, infrared and ultraviolet photometry, optical polarimetry and ultraviolet spectroscopy are included. An invited paper concerning comprehensive theoretical interpretation in the context of stellar evolution also is included. The information collected herein is unparalleled in scope and will remain a standard reference until the next eclipse cycle in the year 2009 A.D., in all probability.

  13. $\\epsilon$-Expansions Near Three Dimensions from Conformal Field Theory

    E-print Network

    Basu, Pallab

    2015-01-01

    We formally extend the CFT techniques introduced in arXiv:1505.00963, to $\\phi^{\\frac{2d_0}{d_0-2}}$ theory in $d=d_0-\\epsilon$ dimensions and use it to compute anomalous dimensions near $d_0=3, 4$ in a unified manner. We also do a similar analysis of the $O(N)$ model in three dimensions by developing a recursive combinatorial approach for OPE contractions. Our results match precisely with low loop perturbative computations. Finally, using 3-point correlators in the CFT, we comment on why the $\\phi^3$ theory in $d_0=6$ is qualitatively different.

  14. Identification of epsilon martensite in a Fe-based shape memory alloy by means of EBSD.

    PubMed

    Verbeken, K; Van Caenegem, N; Raabe, D

    2009-01-01

    Ferrous shape memory alloys (SMAs) are often thought to become a new, important group of SMAs. The shape memory effect in these alloys is based on the reversible, stress-induced martensitic transformation of austenite to epsilon martensite. The identification and quantification of epsilon martensite is crucial when evaluating the shape memory behaviour of this material. Previous work displayed that promising results were obtained when studying the evolution of the amount of epsilon martensite after different processing steps with Electron BackScatter Diffraction (EBSD). The present work will discuss in detail, on the one hand, the challenges and opportunities arising during the identification of epsilon martensite by means of EBSD and, on the other hand, the possible interpretations that might be given to these findings. It will be illustrated that although the specific nature of the austenite to epsilon martensite transformation can still cause some points of discussion, EBSD has a high potential for identifying epsilon martensite. PMID:18280172

  15. Mechanism of Tryptic Activation of Clostridium botulinum Type E Toxin

    PubMed Central

    Gerwing, Julia; Dolman, Claude E.; Ko, Arthur

    1965-01-01

    Gerwing, Julia (University of British Columbia, Vancouver, B.C., Canada), Claude E. Dolman, and Arthur Ko. Mechanism of tryptic activation of Clostridium botulinum type E toxin. J. Bacteriol. 89:1176–1179. 1965.—The toxic peptide of trypsin activated Clostridium botulinum type E toxin was purified by chromatography through columns packed with Sephadex G-75 and G-50. The molecular weight of the active peptide was estimated to lie between 10,000 and 12,000. Amino acid analyses indicated that the active peptide had lost at least 18 of the amino acid residues present in the original protein. The active peptide and the original protein were found to have different N-terminal amino acid residues. The mechanism of tryptic activation apparently involves chiefly the removal of amino acids from the N-terminus of the toxin molecule. PMID:14292982

  16. Nosocomial outbreak of Clostridium difficile diarrhea in a pediatric service.

    PubMed

    Ferroni, A; Merckx, J; Ancelle, T; Pron, B; Abachin, E; Barbut, F; Larzul, J; Rigault, P; Berche, P; Gaillard, J L

    1997-12-01

    An outbreak of nosocomial diarrhea that occurred in a pediatric orthopedic service between 1 December 1993 and 15 April 1994 is reported. A total of 37 patients (mean age, 9.6 years; range, 2 months-19.3 years) were involved in the outbreak, including six patients with bacteriologically documented Clostridium difficile infection. A multivariate analysis identified lincomycin treatment for at least three days as the only significant risk factor. Stool samples from four asymptomatic patients were also positive for Clostridium difficile and its cytotoxins. Isolates from all patients belonged to serogroup C, were highly resistant to lincomycin, and exhibited the same restriction pattern by pulsed-field gel electrophoresis. The outbreak ended after treatment with lincomycin was discontinued and hygiene control measures were implemented. PMID:9495676

  17. Models for the study of Clostridium difficile infection

    PubMed Central

    Best, Emma L.; Freeman, Jane; Wilcox, Mark H.

    2012-01-01

    Models of Clostridium difficile infection (C. difficile) have been used extensively for Clostridium difficile (C. difficile) research. The hamster model of C. difficile infection has been most extensively employed for the study of C. difficile and this has been used in many different areas of research, including the induction of C. difficile, the testing of new treatments, population dynamics and characterization of virulence. Investigations using in vitro models for C. difficile introduced the concept of colonization resistance, evaluated the role of antibiotics in C. difficile development, explored population dynamics and have been useful in the evaluation of C. difficile treatments. Experiments using models have major advantages over clinical studies and have been indispensible in furthering C. difficile research. It is important for future study programs to carefully consider the approach to use and therefore be better placed to inform the design and interpretation of clinical studies. PMID:22555466

  18. Comparison of toxins of Clostridium butyricum and Clostridium botulinum type E.

    PubMed Central

    Giménez, J A; Sugiyama, H

    1988-01-01

    The toxin of Clostridium butyricum strains isolated from two infants with botulism is neutralized by antitoxin for type E botulinum toxin. This toxin and that of a C. botulinum type E strain were purified by the same protocol. Both toxins were Mr 145,000 proteins which, when activated with trypsin, were composed of an H subunit of Mr 105,000 and an L subunit of Mr 50,000. The activated specific toxicity of purified butyricum toxin based on an intravenous assay was 2 X 10(8) mouse 50% lethal doses (LD50s)/mg of protein, but that based on an intraperitoneal assay was 7 X 10(7) LD50s/mg, compared with 6 X 10(7) LD50s/mg for type E toxin as determined by both methods. Immunodiffusion tests with antitoxin raised with type E toxin indicated that the two toxins were serologically very similar except for a spur formed by type E toxin. The close similarities of the two toxins suggest that toxigenic C. butyricum could arise when a wild-type strain, which is normally nontoxigenic, acquires the toxin gene of a C. botulinum type E strain. Images PMID:3126148

  19. Local Fourier transform and epsilon factors joint work with Ahmed Abbes

    E-print Network

    Bannai, Kenichi

    and the product formula: L(F, t) = (F)t-(C¯k,F) L(F , (qt)-1 ). 1 #12;(F) = xC x(Fx). The local epsilon factors xLocal Fourier transform and epsilon factors joint work with Ahmed Abbes July 3, 2008, 17:00-18:00, at the Univ. of Tokyo Abstract The local epsilon factors appear in the constant term of the functional equa

  20. Medical diagnoses and procedures associated with Clostridium difficile colitis

    Microsoft Academic Search

    Anna M Buchner; Amnon Sonnenberg

    2001-01-01

    Objectives:The aim of this study was to examine the associations of Clostridium difficile colitis with other comorbid conditions and procedural interventions among hospitalized patients.Methods:The Patient Treatment File of the Department of Veterans Affairs contains the computerized records of all inpatients treated in 172 Veterans Affairs hospitals distributed throughout the United States. The computerized medical records of 15,091 cases with C.

  1. Medical diagnoses and procedures associated with Clostridium difficile colitis

    Microsoft Academic Search

    Anna M. Buchner; Amnon Sonnenberg

    2001-01-01

    OBJECTIVES:The aim of this study was to examine the associations of Clostridium difficile colitis with other comorbid conditions and procedural interventions among hospitalized patients.METHODS:The Patient Treatment File of the Department of Veterans Affairs contains the computerized records of all inpatients treated in 172 Veterans Affairs hospitals distributed throughout the United States. The computerized medical records of 15,091 cases with C.

  2. Antisense RNA Strategies for Metabolic Engineering of Clostridium acetobutylicum

    Microsoft Academic Search

    RUCHIR P. DESAI; ELEFTHERIOS T. PAPOUTSAKIS

    1999-01-01

    We examined the effectiveness of antisense RNA (as RNA) strategies for metabolic engineering of Clostridium acetobutylicum. Strain ATCC 824(pRD4) was developed to produce a 102-nucleotide asRNA with 87% comple- mentarity to the butyrate kinase (BK) gene. Strain ATCC 824(pRD4) exhibited 85 to 90% lower BK and acetate kinase specific activities than the control strain. Strain ATCC 824(pRD4) also exhibited 45

  3. Conjugative Botulinum Neurotoxin-Encoding Plasmids in Clostridium botulinum

    Microsoft Academic Search

    Kristin M. Marshall; Marite Bradshaw; Eric A. Johnson; Holger Bruggemann

    2010-01-01

    BackgroundClostridium botulinum produces seven distinct serotypes of botulinum neurotoxins (BoNTs). The genes encoding different subtype neurotoxins of serotypes A, B, F and several dual neurotoxin-producing strains have been shown to reside on plasmids, suggesting that intra- and interspecies transfer of BoNT-encoding plasmids may occur. The objective of the present study was to determine whether these C. botulinum BoNT-encoding plasmids are

  4. Characterization of Clostridium sp. RKD producing botulinum-like neurotoxin

    Microsoft Academic Search

    Aparna Dixit; Ram Kumar Dhaked; Syed Imteyaz Alam; Lokendra Singh

    2005-01-01

    A Gram positive, motile, rod-shaped, strictly anaerobic bacterium isolated from intestine of decaying fish was identified as Clostridium sp. RKD and produced a botulinum type B-like neurotoxin as suggested by mouse bioassay and protection with anti botulinum antibodies. The neurotoxicity was functionally characterized by the phrenic nerve hemi-diaphragm assay. Phylogenetic analysis based on 16S rDNA sequence, placed it at a

  5. Clostridium Infections Associated with Musculoskeletal-Tissue Allografts

    Microsoft Academic Search

    Marion A. Kainer; Jeanne V. Linden; David N. Whaley; Harvey T. Holmes; William R. Jarvis; Daniel B. Jernigan; Lennox K. Archibald

    2004-01-01

    background Allografts are commonly used in orthopedic reconstructive surgery. In 2001, approxi- mately 875,000 musculoskeletal allografts were distributed by U.S. tissue banks. After the death from Clostridium sordellii sepsis of a 23-year-old man who had received a con- taminated allograft from a tissue bank (Tissue Bank A), the Centers for Disease Con- trol and Prevention initiated an investigation, including enhanced

  6. Treating Clostridium difficile Infection with Fecal Microbiota Transplantation

    PubMed Central

    Bakken, Johan S.; Borody, Thomas; Brandt, Lawrence J.; Brill, Joel V.; Demarco, Daniel C.; Franzos, Marc Alaric; Kelly, Colleen; Khoruts, Alexander; Louie, Thomas; Martinelli, Lawrence P.; Moore, Thomas A.; Russell, George; Surawicz, Christina

    2011-01-01

    Clostridium difficile infection is increasing in incidence, severity, and mortality. Treatment options are limited and appear to be losing efficacy. Recurrent disease is especially challenging; extended treatment with oral vancomycin is becoming increasingly common but is expensive. Fecal microbiota transplantation (FMT) is safe, inexpensive, and effective; according to case and small series reports, about 90% of patients are cured. We discuss the rationale, methods, and use of FMT. PMID:21871249

  7. Clostridium difficile in Children: To Treat or Not to Treat?

    PubMed Central

    2014-01-01

    Clostridium difficile infection has been increasing since 2000 in children and in adults. Frequent antibiotics use, comorbidity, and the development of hypervirulent strains have increased the risk of infection. Despite the high carriage rates of C. difficile, infants rarely develop clinical infection. Discontinuing antibiotics and supportive management usually leads to resolution of disease. Antibiotics use should be stratified depending on the patient's age and severity of the disease. PMID:25061582

  8. Nosocomial outbreak of Clostridium difficile diarrhea in a pediatric service

    Microsoft Academic Search

    A. Ferroni; J. Merckx; T. Ancelle; B. Pron; E. Abachin; F. Barbut; J. Larzul; P. Rigault; P. Berche; J.-L. Gaillard

    1997-01-01

    An outbreak of nosocomial diarrhea that occurred in a pediatric orthopedic service between 1 December 1993 and 15 April 1994 is reported. A total of 37 patients (mean age, 9.6 years; range, 2 months-19.3 years) were involved in the outbreak, including six patients with bacteriologically documentedClostridium difficile infection. A multivariate analysis identified lincomycin treatment for at least three days as

  9. Clostridium difficile-associated reactive arthritis in two children

    Microsoft Academic Search

    Helga A. Löffler; Benedicte Pron; Richard Mouy; Nico M. Wulffraat; Anne-Marie Prieur

    2004-01-01

    In adults, reactive arthritis (ReA) following Clostridium difficile-enterocolitis has been documented. In children, only one case of C. difficile-associated ReA has been reported. We now describe two other cases of ReA associated with C. difficile in children. The characteristics of ReA due to C. difficile appear to be similar in adults and children. Both children show polyarthritis after an episode of

  10. AChR deficiency due to epsilon-subunit mutations: two common mutations in the Netherlands.

    PubMed

    Faber, Catharina G; Molenaar, Peter C; Vles, Johannes S H; Bonifati, Domenic M; Verschuuren, Jan J G M; van Doorn, Pieter A; Kuks, Jan B M; Wokke, John H J; Beeson, David; De Baets, Marc

    2009-10-01

    Congenital myasthenic syndromes are a clinically and genetically heterogeneous group of hereditary disorders affecting neuromuscular transmission. We have identified mutations within the acetylcholine receptor (AChR) epsilon-subunit gene underlying congenital myasthenic syndromes in nine patients (seven kinships) of Dutch origin. Previously reported mutations epsilon1369delG and epsilonR311Q were found to be common; epsilon1369delG was present on at least one allele in seven of the nine patients, and epsilonR311Q in six. Phenotypes ranged from relatively mild ptosis and external ophthalmoplegia to generalized myasthenia. The common occurrence of epsilonR311Q and epsilon1369delG suggests a possible founder for each of these mutations originating in North Western Europe, possibly in Holland. Knowledge of the ethnic or geographic origin within Europe of AChR deficiency patients can help in targeting genetic screening and it may be possible to provide a rapid genetic diagnosis for patients of Dutch origin by screening first for epsilonR311Q and epsilon1369delG. PMID:19544078

  11. Magnetic field measurements of $\\epsilon$ Eridani from Zeeman broadening

    E-print Network

    Lehmann, L T; Carroll, T A; Strassmeier, K G

    2015-01-01

    We present new magnetic field measurements of the K2 main-sequence star $\\epsilon$ Eridani based on principal components analysis (PCA) line-profile reconstructions. The aim of this paper is to quantify the surface-averaged magnetic field and search for possible variations. A total of 338 optical echelle spectra from our robotic telescope facility STELLA with a spectral resolution of 55,000 were available for analysis. This time-series was used to search for the small line-profile variations due to a surface magnetic field with the help of a PCA. Evidence for a spatial and temporal inhomogeneous magnetic field distribution is presented. The mean, surface averaged, magnetic field strength was found to be $\\langle B \\rangle = 186 \\pm 47\\,\\mathrm{G}$ in good agreement with previous Zeeman-broadening measurements. Clear short-term variations of the surface averaged magnetic field of up to few tens Gauss were detected together with evidence for a three-year cycle in the surface-averaged magnetic field of $\\epsilon...

  12. Clostridium difficile Colitis Associated with Infant Botulism: Near?Fatal Case Analogous to Hirschsprung's Enterocolitis

    Microsoft Academic Search

    Robert Schechter; Bradley Peterson; James McGee; Olajire Idowu; John Bradley

    1999-01-01

    We present the first five reported cases of Clostridium difficile-associated diarrhea (CDAD) in children with infant botulism caused by Clostridium botulinum. We compare two fulminant cases of colitis in children with colonic stasis, the first caused by infant botulism and the second caused by Hirschsprung's disease. In both children, colitis was accompanied by hypovolemia, hypotension, profuse ascites, pulmonary effusion, restrictive

  13. Ribotyping as an identification tool for Clostridium botulinum strains causing human botulism

    Microsoft Academic Search

    Sebastian Hielm; Johanna Björkroth; Eija Hyytiä; Hannu Korkeala

    1999-01-01

    Ribotyping was used for characterisation of 68 Clostridium botulinum strains and five related Clostridium species to determine the applicability of this method for identification of species causing human botulism. Thirteen restriction enzymes were initially tested for suitability for ribotyping of C. botulinum, of which EcoRI and HindIII were selected. Both enzymes clearly differentiated between proteolytic (group I) and a nonproteolytic

  14. Phylogenetic analysis of Clostridium botulinum type A by multi-locus sequence typing

    Microsoft Academic Search

    Mark J. Jacobson; Guangyun Lin; Thomas S. Whittam; Eric A. Johnson

    2008-01-01

    The genus Clostridium comprises a heterogeneous group of organisms for which the phylogeny and evolutionary relationships are poorly understood. The elucidation of these evolutionary relationships necessitates the use of experimental methods that can distinguish Clostridium lineages that are time and cost effective, and can be accurately and reproducibly employed in different laboratories. Multi-locus sequence typing (MLST) has been successfully used

  15. Genome Sequence of Clostridium tunisiense TJ, Isolated from Drain Sediment from a Pesticide Factory

    PubMed Central

    Sun, Lili; Wang, Yu; Yu, Chunyan; Zhao, Yongqin

    2012-01-01

    Clostridium tunisiense is a Gram-positive, obligate anaerobe that was first isolated in an anaerobic evironment under eutrophication. Here we report the first genome sequence of the Clostridium tunisiense TJ isolated from drain sediment of a pesticide factory in Tianjin, China. The genome is of great importance for both basic and application research. PMID:23209212

  16. Draft Genome Sequence of Antimicrobial-Producing Clostridium sp. JC272, Isolated from Marine Sediment

    PubMed Central

    Tushar, L.; Sasi Jyothsna, T. S.; Ramana, C. V.

    2015-01-01

    We announce the draft genome sequence of Clostridium sp. JC272, isolated from a sediment sample collected from marine habitats of Gujarat, India. Clostridium sp. JC272 is an obligate anaerobe and has the ability to produce antimicrobial compounds. The genome sequence indicates the strain’s capability of producing small peptides (microcins), which are potential novel antibiotics. PMID:26067979

  17. The Strong Isospin-Breaking Correction for the Gluonic Penguin Contribution to epsilon'/epsilon at Next-to-Leading Order in the Chiral Expansion

    E-print Network

    C. E. Wolfe; K. Maltman

    2000-07-27

    The strong isospin-breaking correction, Omega_{st}, which appears in estimates of the Standard Model value for the direct CP-violating ratio epsilon'/epsilon, is evaluated to next-to-leading order (NLO) in the chiral expansion using Chiral Perturbation Theory. The relevant linear combinations of the unknown NLO CP-odd weak low-energy constants (LEC's) which, in combination with 1-loop and strong LEC contributions, are required for a complete determination at this order, are estimated using two different models. It is found that, to NLO, Omega_{st}=0.08 +/- 0.05, significantly reduced from the ``standard'' value, 0.25 +/- 0.08, employed in recent analyses. The potentially significant numerical impact of this decrease on Standard Model predictions for epsilon'/epsilon, associated with the decreased cancellation between gluonic penguin and electroweak penguin contributions, is also discussed.

  18. Results on K --> 2pi decays at O(p^6) and epsilon'/epsilon from an effective chiral lagrangian approach

    E-print Network

    A. A. Bel'kov; G. Bohm; A. V. Lanyov; A. A. Moshkin

    1997-08-20

    We have combined a new systematic calculation of mesonic matrix elements at O(p^6) from an effective chiral lagrangian approach using Wilson coefficients taken from [G.Buchalla, A.J.Buras, M.E.Lautenbacher, Rev.Mod.Phys. 68 (1996) 1125], derived in the framework of perturbative QCD, and restricted partly by experimental data. We derive complete expressions for K --> 2pi amplitudes and give new estimates for epsilon'/epsilon.

  19. Middle atmosphere electrical structure during MAC/EPSILON

    NASA Technical Reports Server (NTRS)

    Mitchell, J. D.; Croskey, C. L.; Blood, S. P.; Li, C.; Hale, L. C.; Goldberg, R. A.

    1990-01-01

    Extensive use of rocket-launched probes during the the MAC/EPSILON campaign at the Andoya Rocket Range, Norway, has enabled the characterization of the region's electrical environment for all four flight series. The first rocket salvo was conducted during daylight (October 15, 1987) and the subsequent three occurred at night (October 21 and 28 and November 12, 1987), all of them during geomagnetically disturbed conditions. Measurements of polar electrical conductivity, ion mobility, and number density are presented, and their associated structure is investigated for local auroral ionization effects. This is believed to be the first time that Gerdien condenser mobility measurements have indicated a heavy-ion presence (positively charged aerosols) in the auroral mesopause region.

  20. Shock initiation of an {epsilon}-CL-20-estane formulation

    SciTech Connect

    Tarver, C.M.; Simpson, R.L.; Urtiew, P.A. [Lawrence Livermore National Laboratory, P.O. Box 808, L-282, Livermore, California 94551 (United States)

    1996-05-01

    The shock sensitivity of a pressed solid explosive formulation, LX-19, containing 95.2{percent} by weight epsilon phase 2,4,6,8,10,12-hexanitrohexaazaisowurtzitane (HNIW) and 4.8{percent} Estane binder, was determined using the wedge test and embedded manganin pressure gauge techniques. This formulation was shown to be slightly more sensitive than LX-14, which contains 95.5{percent} HMX and 4.5{percent} Estane binder. The measured pressure histories for LX-19 were very similar to those obtained using several HMX-inert binder formulations. An Ignition and Growth reactive flow model for LX-19 was developed which differed from those for HMX-inert binder formulations only by a 25{percent} higher hot spot growth rate. {copyright} {ital 1996 American Institute of Physics.}

  1. Goos-Hänchen effect in epsilon-near-zero metamaterials

    PubMed Central

    Xu, Yadong; Chan, C. T.; Chen, Huanyang

    2015-01-01

    Light reflection and refraction at an interface between two homogeneous media is analytically described by Snell's law. For a beam with a finite waist, it turns out that the reflected wave experiences a lateral displacement from its position predicted by geometric optics. Such Goos-Hänchen (G-H) effect has been extensively investigated among all kinds of optical media, such as dielectrics, metals, photonic crystals and metamaterials. As a fundamental physics phenomenon, the G-H effect has been extended to acoustics and quantum mechanics. Here we report the unusual G-H effect in zero index metamaterials. We show that when linearly polarized light is obliquely incident from air to epsilon-near-zero metamaterials, no G-H effect could be observed for p polarized light. While for s polarization, the G-H shift is a constant value for any incident angle. PMID:25731726

  2. Goos-Hänchen effect in epsilon-near-zero metamaterials

    NASA Astrophysics Data System (ADS)

    Xu, Yadong; Chan, C. T.; Chen, Huanyang

    2015-03-01

    Light reflection and refraction at an interface between two homogeneous media is analytically described by Snell's law. For a beam with a finite waist, it turns out that the reflected wave experiences a lateral displacement from its position predicted by geometric optics. Such Goos-Hänchen (G-H) effect has been extensively investigated among all kinds of optical media, such as dielectrics, metals, photonic crystals and metamaterials. As a fundamental physics phenomenon, the G-H effect has been extended to acoustics and quantum mechanics. Here we report the unusual G-H effect in zero index metamaterials. We show that when linearly polarized light is obliquely incident from air to epsilon-near-zero metamaterials, no G-H effect could be observed for p polarized light. While for s polarization, the G-H shift is a constant value for any incident angle.

  3. Goos-Hänchen effect in epsilon-near-zero metamaterials.

    PubMed

    Xu, Yadong; Chan, C T; Chen, Huanyang

    2015-01-01

    Light reflection and refraction at an interface between two homogeneous media is analytically described by Snell's law. For a beam with a finite waist, it turns out that the reflected wave experiences a lateral displacement from its position predicted by geometric optics. Such Goos-Hänchen (G-H) effect has been extensively investigated among all kinds of optical media, such as dielectrics, metals, photonic crystals and metamaterials. As a fundamental physics phenomenon, the G-H effect has been extended to acoustics and quantum mechanics. Here we report the unusual G-H effect in zero index metamaterials. We show that when linearly polarized light is obliquely incident from air to epsilon-near-zero metamaterials, no G-H effect could be observed for p polarized light. While for s polarization, the G-H shift is a constant value for any incident angle. PMID:25731726

  4. High-Pressure Structural Study of Epsilon HNIW (CL-20)

    NASA Astrophysics Data System (ADS)

    Gump, Jared C.; Wong, Chak P.; Zerilli, Frank J.; Peiris, Suhithi M.

    2004-07-01

    The structure of epsilon CL-20 at room temperature was investigated using synchrotron angle-dispersive x-ray diffraction experiments and Raman spectroscopy. For x-ray diffraction, the samples were compressed up to 6.3 GPa using a Merrill-Bassett diamond anvil cell (DAC) under both hydrostatic and non-hydrostatic conditions. Pressure — volume data were then fit to the Birch-Murnaghan equation of state to obtain an isothermal equation of state. No phase transition was observed within this pressure range. Raman spectroscopy was performed in the range of 50-1650 cm-1. The samples were compressed non-hydrostatically to 7.1 GPa. Changes in peak positions with increasing pressure were observed. Vibrational spectra were calculated using Hartree-Fock and density functional theory and a comparison was made with the experimental spectrum.

  5. Electric Levitation Using Epsilon-Near-Zero Metamaterials

    E-print Network

    Fortuño, Francisco J Rodríguez; Engheta, Nader

    2013-01-01

    Levitation of objects with action at a distance has always been intriguing to humans. Several ways to achieve this, such as aerodynamic, acoustic, or electromagnetic methods, including radiation pressure, stable potential wells, and quantum Casimir-Lifshitz forces, exist. A fascinating approach for levitation is that of magnets over superconductors based on the Meissner effect -the expulsion of the magnetic field by a superconductor. With the advent of metamaterials -designed structures with electromagnetic properties that may not be found in nature- we ask whether a material may be conceived exhibiting similar field expulsion, but involving the electric field. We show how a special subcategory of metamaterials, called epsilon-near-zero materials, exhibits such electric classic analog to the Meissner effect, exerting a repulsion on nearby sources. Repulsive forces using anisotropic and chiral metamaterials have been investigated, but our proposal uses a different mechanism based on field expulsion, and is ver...

  6. Compressive Epsilon Photography for Post-Capture Control in Digital Imaging Atsushi Ito1

    E-print Network

    Mellor-Crummey, John

    Categories: I.2.10 [Artificial Intelligence]: Vision and Scene Understanding--3D/stereo scene analysis I.4Compressive Epsilon Photography for Post-Capture Control in Digital Imaging Atsushi Ito1 , Salil Rice University 3 Carnegie Mellon University Figure 1: Compressive epsilon photography enables post

  7. An efficient, adaptive parameter variation scheme for metaheuristics based on the epsilon-constraint method

    E-print Network

    Zitzler, Eckart

    An efficient, adaptive parameter variation scheme for metaheuristics based on the epsilon value a priori is shown to be a serious drawback of the original epsilon-constraint method. We therefore ), where k is the number of Pareto-optimal solutions to be found and m the number of objectives. Simulation

  8. LOCAL EPSILON ISOMORPHISMS DAVID LOEFFLER, OTMAR VENJAKOB, AND SARAH LIVIA ZERBES

    E-print Network

    Venjakob, Otmar

    LOCAL EPSILON ISOMORPHISMS DAVID LOEFFLER, OTMAR VENJAKOB, AND SARAH LIVIA ZERBES Abstract. Comparison with Kato's rank one epsilon-isomorphisms 50 Acknowledgements 51 References 51 1. Introduction 1 of those): via the usual functorial properties of determinants, our results im- ply the conjectures CIW (K

  9. A comparison of the linear and nonlinear k-epsilon turbulence models in combustors

    Microsoft Academic Search

    C. C. Hwang; Genxing Zhu; M. Massoudi; J. M. Ekmann

    1993-01-01

    The nonlinear k-epsilon closure is presently used to model the turbulence in combustor geometries involving (1) the flow in a round pipe entering an expansion into another coaxial round pipe, and (2) the flow in two confined coaxial swirling jets. No significant differences are discernible in the performance of the two models; the inlet conditions for k and epsilon are

  10. Modification of k-epsilon turbulence models for coaxial sudden-expansion swirling flows

    Microsoft Academic Search

    Hau-Chung Soong; Keh-Chin Chang

    1989-01-01

    A successful prediction of turbulent swirling flow, which is important for simulation of gas turbine combustors and ramjet combustors, requires a reliable turbulence model. The standard k - epsilon model, which is commonly used for engineering practice, is reported in less satisfactory agreement with the measured results for flows incorporated with high streamline curvature effects. Several modified k - epsilon

  11. A modified form of the k-epsilon model for predicting wall turbulence

    Microsoft Academic Search

    C. K. G. Lam; K. Bremhorst

    1981-01-01

    The high Reynolds number form of the k-epsilon model is extended and tested by application to a fully developed pipe flow. It is established that the model is valid throughout the fully turbulent, semilaminar and laminar regions of the flow. Unlike many previously proposed forms of the k-epsilon model, the present form does not have to be used in conjunction

  12. On the realisation of maximal simple types and epsilon factors of pairs

    E-print Network

    Stevens, Shaun

    On the realisation of maximal simple types and epsilon factors of pairs Vytautas Paskunas and Shaun the epsilon factor, using the definition of Jacquet, Piatetskii-Shapiro, Shalika. We also investigate . . . . . . . . . . 32 4.4 A characterisation of Ind K(A) J . . . . . . . . . . . . . . . . . . 33 5 Realisation

  13. The Hypergeometric Representation of Feynman Diagrams and Construction of the Epsilon Expansion

    E-print Network

    Yost, Scott

    The Hypergeometric Representation of Feynman Diagrams and Construction of the Epsilon Expansion S the epsilon expansion S.A. Yost Hypergeometric Representation of Feynman Diagrams A.M.S. Meeting, New Orleans lines (labeled k) is unconstrained and momentum conservation implies q1 = p1 - k, q2 = p1 + p2 - k, q3

  14. Testing a low-Reynolds number k-epsilon turbulence model based on direct simulation data

    Microsoft Academic Search

    V. Michelassi; W. Rodi; J. Zhu

    1993-01-01

    A low Re k-epsilon model based on direct numerical simulation (DNS) data proposed by Rodi and Mansour (1993) (RM) is discussed. The model was complemented by a damping function multiplying the destruction term in the model epsilon equation. The modified version of the RM model yielded generally good predictions of all major quantities in developed channel and boundary layer flows

  15. Sequential crowdsourced labeling as an epsilon-greedy exploration in a Markov Decision Process

    E-print Network

    Kaski, Samuel

    Sequential crowdsourced labeling as an epsilon-greedy exploration in a Markov Decision Process one at a time. We model it as an epsilon-greedy exploration in a Markov Decision Process. In order to get multiple labels the task requester specifies that each HIT has to be completed by k workers

  16. Differential regulation of alternative 3{prime} splicing of {epsilon} messenger RNA variants

    SciTech Connect

    Diaz-Sanchez, D.; Zhang, K.; Saxon, A. [Univ. of California School of Medicine, Los Angeles, CA (United States)] [and others

    1995-08-15

    Alternative 3{prime} splicing of the one active human {epsilon} heavy chain gene results in variants of {epsilon} mRNA encoding distinct IgE proteins. The same relative amounts of these {epsilon} mRNA variants were produced by non-atopic donor B cells when driven in a variety of T-dependent or T-independent systems. The most abundant variants were those for classic secreted {epsilon} and a novel secreted form (CH4-M2{double_prime}). In contrast, cells from subjects with high levels of serum IgE secondary to parasitic infection or atopy spontaneously produced higher relative levels of the CH4-M2{prime} {epsilon} mRNA variant, lower relative amounts of both the membrane and CH4-M2{double_prime} secreted variants, and very low levels of the CH4{prime}-CH5 variant. The existence of and corresponding changes in levels of the CH4-M2{prime}-enclosed secreted protein were demonstrated. IL-10 induced this same differential expression of {epsilon} splice variants in vitro when used to costimulate IL-4 plus CD40-driven B cells and could differentially enhance the production of CH4-M2{prime} protein by established IgE-secreting cell lines. Inhibition of IgE by cross-linking the low affinity IgE receptor (CD23) decreased the levels of {epsilon} mRNA and resulted in a distinct pattern of {epsilon} mRNA characterized by a dramatic decrease in CH4-M2{prime} splice variant. IL-6, IL-2, or IFN-{gamma} did not change the {epsilon} mRNA pattern. Overall, the absolute and relative amounts of the different {epsilon} mRNA splice variants produced appear to be controlled in a differentiation-related fashion.

  17. Delta I=1/2 Rule, epsilon'/epsilon and K -> pi nu nubar in Z'(Z) and G' Models with FCNC Quark Couplings

    E-print Network

    Andrzej J. Buras; Fulvia De Fazio; Jennifer Girrbach

    2014-07-09

    The experimental value for the isospin amplitude Re(A_2) in K->pi pi decays has been successfully explained within the Standard Model, both within large N approach to QCD and by QCD lattice calculations. On the other hand in both approaches the theoretical values of Re(A_0) are by at least 30% below the data so that the Delta I=1/2 rule in K->pi pi decays is not fully explained. While this deficit could be the result of present theoretical uncertainties in both approaches, it cannot be excluded that the missing piece in Re(A_0) comes from New Physics. We demonstrate that this deficit can be significantly softened by tree-level FCNC transitions mediated by a heavy colourless Z' gauge boson with particularly chosen flavour violating couplings assuring negligible NP contributions to Re(A_2) and significantly enhancing the contribution of the leading QCD penguin operator Q_6 to Re(A_0). A large fraction of the missing piece in the Delta I=1/2 rule can be explained in this manner for M_{Z'} in the reach of the LHC, while satisfying constraints from epsilon_K, epsilon'/epsilon, Delta M_K, LEP-II and the LHC. We identify quartic correlation between Z' contributions to Re(A_0), epsilon'/epsilon, epsilon_K and Delta M_K. We present correlations between epsilon'/epsilon and rare decays K->pi nu bar{nu} with and without the Delta I=1/2 rule constraint and generalize the whole analysis to Z' with colour (G') and Z with FCNC couplings. In the latter case no improvement on Re(A_0) can be achieved without destroying the agreement of the SM with the data on Re(A_2). Moreover, this scenario is very tightly constrained by epsilon'/epsilon. On the other hand in the context of the Delta I=1/2 rule G' is even more effective than Z': it provides the missing piece in Re(A_0) for M_{G'}=(3.5-4.0) TeV.

  18. New advances in the treatment of Clostridium difficile infection (CDI)

    PubMed Central

    Hedge, Dennis D; Strain, Joe D; Heins, Jodi R; Farver, Debra K

    2008-01-01

    Clostridium difficile infections (CDI) have increased in frequency throughout the world. In addition to an increase in frequency, recent CDI epidemics have been linked to a hypervirulent C. difficile strain resulting in greater severity of disease. Although most mild to moderate cases of CDI continue to respond to metronidazole or vancomycin, refractory and recurrent cases of CDI may require alternative therapies. This review provides a brief overview of CDI and summarizes studies involving alternative antibiotics, toxin binders, probiotics, and immunological therapies that can be considered for treatment of acute and recurrent CDI in severe and refractory situations. PMID:19209277

  19. Transcriptional Analysis of Butanol Stress and Tolerance in Clostridium acetobutylicum

    Microsoft Academic Search

    Christopher A. Tomas; Jeffrey Beamish; Eleftherios T. Papoutsakis

    2006-01-01

    The effects of challenges with low (0.25%, vol\\/vol) and high (0.75%) concentrations of butanol on the growth, glucose metabolism, product formation, and transcriptional program of the solvent-tolerant Clostridium ace- tobutylicum strain 824(pGROE1) and the plasmid control strain 824(pSOS95del) were used to study solvent tolerance and stress response. Strain 824(pGROE1) was generated by groESL overexpression. The growth of 824(pGROE1) was less

  20. Pseudomembranous Colitis: Not Always Caused by Clostridium difficile

    PubMed Central

    Tang, Derek M.; Urrunaga, Nathalie H.; De Groot, Hannah; Ghazi, Leyla J.

    2014-01-01

    Although classically pseudomembranous colitis is caused by Clostridium difficile, it can result from several etiologies. Certain medications, chemical injury, collagenous colitis, inflammatory bowel disease, ischemia, and other infectious pathogens can reportedly cause mucosal injury and subsequent pseudomembrane formation. We present the case of a middle-aged woman with vascular disease who was incorrectly diagnosed with refractory C. difficile infection due to the presence of pseudomembranes. Further imaging, endoscopy, and careful histopathology review revealed chronic ischemia as the cause of her pseudomembranous colitis and diarrhea. This case highlights the need for gastroenterologists to consider non-C. difficile etiologies when diagnosing pseudomembranous colitis. PMID:25214850

  1. Structure, Function and Regulation of the Clostridium cellulovorans Cellulosome

    SciTech Connect

    Doi, Roy H

    2008-06-01

    Our major goal for this project (2004-2008) was to obtain an understanding ofthe structure, function, and regulation of the Clostridium cellulovorans cellulosomes. Our specific goals were to select genes for cellulosomal and non-cellulosomal enzymes and characterize their products, to study the synergistic action between cellulosomal and non-cellulosomal enzymes, to study the composition of cellulosomes when cells were grown with different carbon sources, continue our studies on the scaffolding protein and examine heterologous expression of cellulosomal genes in Bacillus subtilis. We fulfilled the specific goals of our proposal.

  2. Antibacterial functionalization of wool via mTGase-catalyzed grafting of epsilon-poly-L-lysine.

    PubMed

    Wang, Qiang; Jin, Guibiao; Fan, Xuerong; Zhao, Xianfei; Cui, Li; Wang, Ping

    2010-04-01

    epsilon-Poly-L-lysine (epsilon-PL), a natural biomacromolecule having a broad spectrum of antibacterial activity, was grafted on the wool fiber via the acyl transfer reaction catalyzed by microbial transglutaminase (mTGase) to develop a new strategy for antibacterial functionalization of proteinous materials. The effects of the concentrations of epsilon-PLs and mTGases on the graft yields were investigated. A coating of epsilon-PL that almost completely covered the scale profile on the wool surface was visualized by scanning electron microscopy (SEM) and further demonstrated in terms of Allwörden's reaction characteristic of wool. Identifiable differences in lysine content and color depth among the stained wool samples reveal the changes in the surface composition and polarity caused by the incorporation of epsilon-PL onto the wool substrate, respectively. The ratio of bacteriostasis to Escherichia coli of the wool fabric grafting epsilon-PL reached 96.6 %, indicating an excellent antibacterial effect. The application of epsilon-PL and corresponding mTGase-catalyzed grafting reaction would provide a worthwhile reference for antibacterial functionalization of proteinous materials in various forms. PMID:19649747

  3. Detection of cold-tolerant clostridia other than Clostridium estertheticum in raw vacuum-packed chill-stored meat.

    PubMed

    Cavill, Laura; Renteria-Monterrubio, Ana L; Helps, Christopher R; Corry, Janet E L

    2011-08-01

    Samples from raw chill-stored vacuum-packed beef, lamb and venison or the meat processing environment, associated with a spoilage problem, but negative for Clostridium estertheticum using a specific real-time PCR test, were examined for other Clostridium spp. using direct 16S rDNA PCR-RFLP and sequencing. Of 291 samples tested by PCR, presence of clostridia was indicated in 123 and there was sufficient PCR product in 35 to be further investigated. Presence of Clostridium spp. was confirmed by RFLP and sequencing in 25/35 samples (11 of 14 incidents). Species detected in spoiled meat were (incidents): Clostridium tagluense-like (4), Clostridium putrefaciens (2), Clostridium algidicarnis (3), Clostridium frigoris/estertheticum-like (3) and Clostridium. gasigenes (2). More than one species was detected in some incidents. All of the above species have previously been associated with spoiled meat apart from the Cl. tagluense-like species. Clostridia were also confirmed in 4/7 samples from the environment, with two Cl. frigoris/estertheticum-like and two mesophilic species of Clostridium. Our study showed that, cold-tolerant Clostridium species other than Cl. estertheticum are occasionally associated with spoiled vacuum-packed meat, particularly lamb. Further studies are required to confirm the exact identity of the Cl. tagluense-like species and its role in meat spoilage. PMID:21569939

  4. CRYSTAL STRUCTURE OF CLOSTRIDIUM BOTULINUM NEUROTOXIN SEROTYPE B.

    SciTech Connect

    SWAMINATHAN,S.; ESWARAMOORTHY,S.

    2001-11-19

    The toxigenic strains of Clostridium botulinum produce seven serologically distinct types of neurotoxins labeled A - G (EC 3.4.24.69), while Clostridium tetani produces tetanus neurotoxin (EC 3.4.24.68). Botulinum and tetanus neurotoxins (BoNTs and TeNT) are produced as single inactive chains of molecular mass of approximately 150 kDa. Most of these neurotoxins are released after being cleaved into two chains, a heavy chain (HI) of 100 kDa and a light chain (L) of 50 kDa held together by an interchain disulfide bond, by tissue proteinases. BoNT/E is released as a single chain but cleaved by host proteinases [1]. Clostvidium botulinum neurotoxins are extremely poisonous proteins with their LD{sub 50} for humans in the range of 0.1 - 1 ng kg{sup -1} [2]. Botulinum neurotoxins are responsible for neuroparalytic syndromes of botulism characterized by serious neurological disorders and flaccid paralysis. BoNTs block the release of acetylcholine at the neuromuscular junction causing flaccid paralysis while TeNT blocks the release of neurotransmitters like glycine and {gamma}-aminobutyric acid (GABA) in the inhibitory interneurons of the spinal cord resulting in spastic paralysis. In spite of different clinical symptoms, their aetiological agents intoxicate neuronal cells in the same way and these toxins have similar structural organization [3].

  5. Characterization of the cellulolytic complex (cellulosome) of Clostridium acetobutylicum.

    PubMed

    Sabathé, Fabrice; Bélaïch, Anne; Soucaille, Philippe

    2002-11-19

    A large cellulosomal gene cluster was identified in the recently sequenced genome of Clostridium acetobutylicum ATCC 824. Sequence analysis revealed that this cluster contains the genes for the scaffolding protein CipA, the processive endocellulase Cel48A, several endoglucanases of families 5 and 9, the mannanase Man5G, and a hydrophobic protein, OrfXp. Surprisingly, genetic organization of this large cluster is very similar to that of Clostridium cellulolyticum, the model of mesophilic clostridial cellulosomes. As C. acetobutylicum is unable to grow on cellulosic substrates, the existence of a cellulosomal gene cluster in the genome raises questions about its expression, function and evolution. Biochemical evidence for the expression of a cellulosomal protein complex was investigated. The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, N-terminal sequencing and Western blotting with antibodies against specific components of the C. cellulolyticum cellulosome suggest that at least four major cellulosomal proteins are present. In addition, despite the fact that no cellulolytic activities were detected, we report here the evidence for the production of a high molecular mass cellulosomal complex in C. acetobutylicum. PMID:12445640

  6. The polar lipids of Clostridium psychrophilum, an anaerobic psychrophile

    PubMed Central

    Guan, Ziqiang; Tian, Bing; Perfumo, Amedea; Goldfine, Howard

    2013-01-01

    We have examined the polar lipids of Clostridium psychrophilum, a recently characterized psychrophilic Clostridium isolated from an Antarctic microbial mat. Lipids were extracted from cells grown near the optimal growth temperature (+5 °C) and at ?5 °C, and analyzed by two-dimensional thin layer chromatography and liquid chromatography coupled with mass spectrometry. The major phospholipids of this species are: cardiolipin, phosphatidylethanolamine, and phosphatidylglycerol. Phosphatidylserine and lyso-phosphatidylethanolamine were found as minor components. The most abundant glycolipids are a monoglycosyldiradylglycerol (MGDRG) and a diglycosyldiradylglycerol (DGDRG). The latter was only seen in cells grown at ?5 °C. An ethanolamine-phosphate derivative of N-acetylglucosaminyldiradylglycerol was seen in cells grown at ?5 °C and an ethanolamine-phosphate derivative of MGDRG was found in cells grown at +5 °C. All lipids were present in both the all acyl and plasmalogen (alk-1?-enyl acyl) forms with the exception of PS and MGDRG, which were predominantly in the diacyl form. The significance of lipid changes at the two growth temperatures is discussed. PMID:23454375

  7. Conserved Oligopeptide Permeases Modulate Sporulation Initiation in Clostridium difficile

    PubMed Central

    Edwards, Adrianne N.; Nawrocki, Kathryn L.

    2014-01-01

    The anaerobic gastrointestinal pathogen Clostridium difficile must form a metabolically dormant spore to survive in oxygenic environments and be transmitted from host to host. The regulatory factors by which C. difficile initiates and controls the early stages of sporulation in C. difficile are not highly conserved in other Clostridium or Bacillus species. Here, we investigated the role of two conserved oligopeptide permeases, Opp and App, in the regulation of sporulation in C. difficile. These permeases are known to positively affect sporulation in Bacillus species through the import of sporulation-specific quorum-sensing peptides. In contrast to other spore-forming bacteria, we discovered that inactivating these permeases in C. difficile resulted in the earlier expression of early sporulation genes and increased sporulation in vitro. Furthermore, disruption of opp and app resulted in greater virulence and increased the amounts of spores recovered from feces in the hamster model of C. difficile infection. Our data suggest that Opp and App indirectly inhibit sporulation, likely through the activities of the transcriptional regulator SinR and its inhibitor, SinI. Taken together, these results indicate that the Opp and App transporters serve a different function in controlling sporulation and virulence in C. difficile than in Bacillus subtilis and suggest that nutrient availability plays a significant role in pathogenesis and sporulation in vivo. This study suggests a link between the nutritional status of the environment and sporulation initiation in C. difficile. PMID:25069979

  8. Biodegradation of trinitrotoluene (TNT) by a strain of Clostridium bifermentans

    SciTech Connect

    Shin, C.Y.; Crawford, D.L. [Univ. of Idaho, Moscow, ID (United States)

    1995-12-31

    A Clostridium capable of degrading 2,4,6-trinitrotoluene (TNT) cometabolically was isolated from a mixed culture obtained from a bioreactor fed TNT. This bacterium, identified as a strain of Clostridium bifermentans, and designated strain CYS-1, was able to degrade TNT via 4-amino-2,6-dinitrotoluene (4-ADNT) and 2,4-diamino-6-nitrotoluene (2,4-DANT) to aliphatic polar products which are now being identified and are assumed to be organic acids. CYS 1 cells are tolerant of TNT and capable of degrading it at starting concentrations of up to {ge}100 mg/L TNT. The number of cells inoculated and the availability of cosubstrate nutrients are significant factors influencing TNT degradation, as are TNT tolerance and survival of the cells at high TNT concentrations. In liquid media, at high TNT concentrations, TNT toxicity could be overcome by increasing the amount of inoculum and supplementing the culture with appropriate rich organic cosubstrates. Under these conditions, the reduction of 4-ADNT to 2,4-DANT occurred very fast, whereas the further degradation of 2,4-DANT proceeded more slowly.

  9. Clostridium difficile colitis: review of the therapeutic approach.

    PubMed

    Joseph, Josmi; Singhal, Shashideep; Patel, Gia M; Anand, Sury

    2014-01-01

    Clostridium difficile infection (CDI) is the leading cause of antibiotic-associated and nosocomial infectious diarrhea. Presenting as clostridium difficile colitis, it is a significant cause of morbidity and mortality. Metronidazole is regarded as the agent of choice for CDl therapy and also for the first recurrence in most patients with mild to moderate CDI. Vancomycin is recommended as an initial therapy for patients with severe CDI. With recent Food and Drug Administration-approval fidaxomicin is available for clinical use and is as effective as vancomycin with lower relapse rates. Rifaximin and fecal bacteriotherapy are alternative approaches in patients with severe or refractory CDI, before surgical intervention. Antibiotic research is ongoing to add potential new drugs such as teicoplanin, ramoplanin, fusidic acid, nitazoxanide, rifampin, bacitracin to our armamentarium. Role of toxin-binding agents is still questionable. Monoclonal antibody and intravenous immunoglobulin are still investigational therapies that could be promising options. The ongoing challenges in the treatment of CDI include management of recurrence and presence of resistance strains such as NAP1/BI/027, but early recognition of surgical candidates can potentially decrease mortality in CDI. PMID:22990077

  10. Simulations of free shear layers using a compressible k-epsilon model

    NASA Technical Reports Server (NTRS)

    Yu, S. T.; Chang, C. T.; Marek, C. J.

    1991-01-01

    A two-dimensional, compressible Navier-Stokes equations with a k-epsilon turbulence model are solved numerically to simulate the flows of compressible free shear layers. The appropriate form of k and epsilon equations for compressible flows are discussed. Sarkar's modeling is adopted to simulate the compressibility effects in the k and epsilon equations. The numerical results show that the spreading rate of the shear layers decreases with increasing convective Mach number. In addition, favorable comparison was found between the calculated results and Goebel and Dutton's experimental data.

  11. Heavy Meson Masses in the \\epsilon-Regime of HM\\chi PT

    E-print Network

    Briceño, Raúl A

    2011-01-01

    The pseudoscalar and vector heavy meson masses are calculated in the \\epsilon-regime of Heavy Meson Chiral Perturbation Theory to order \\epsilon^4. The results of this calculation will allow the determination of low-energy coefficients (LECs) directly from Lattice QCD calculations of the heavy mesons masses for lattices that satisfy the \\epsilon-regime criteria. In particular, the LECs that parametrize the NLO volume dependance of the heavy meson masses are necessary for evaluating the light pseudoscalar meson (\\pi, K, \\eta) and heavy meson ({D^0, D^+, D^+_s}, {B^-,\\bar{B}^0,\\bar{B}^0_s}) scattering phase shifts.

  12. Quantum plasmon effects in epsilon-near-zero metamaterials

    E-print Network

    Moaied, M; Ostrikov, K

    2014-01-01

    Dispersion properties of metals and propagation of quantum bulk plasmon in the high photon energy regime are studied. The nonlocal dielectric permittivity of a metal is determined by the quantum plasma effects and is calculated by applying the Wigner equation in the kinetic theory and taking into account the electron lattice collisions. The properties of epsilon near zero material are investigated in a thin gold film. The spectrum and the damping rate of the quantum bulk plasmon are obtained for a wide range of energies, and the electron wave function is analytically calculated in both classical and quantum limits. It is shown that the quantum bulk plasmons exist with a propagation length of 1 to 10nm, which strongly depends on the electron energy. The propagation length is found to be much larger than the propagation length in the classical regime which is comparable to the atomic radius and the average inter particle distance. It is found that the spatial localization of the electron wave function is extend...

  13. Discovery of the February Epsilon Virginids (FEV, IAU#506)

    NASA Astrophysics Data System (ADS)

    Steakley, Kathryn; Jenniskens, P. M.

    2013-01-01

    Halley type comets are relatively few, but at Earth they are sampled over a large part of the inner solar system because dust accumulates in comparatively stable orbits. We have detected a new meteor shower with a Halley-type orbit, the February epsilon Virginids (FEV), from video observations with the Cameras for All-Sky Meteor Surveillance (CAMS) and by examining orbits listed in the SonataCo Japanese database. Twenty-two meteors were detected during the period from February 1st through February 9th of 2008 to 2012 that are part of this shower. The FEVs originate from the geocentric radiant of R.A. = 201.66° and Dec = +10.39° with a mean geocentric velocity of 63.01 km/s. The mean orbital elements of these meteoroids are q = (0.488 ± 0.021) AU, 1/a = ( 0.085 ± 0.095) 1/AU, e = 0.958 ± 0.046, i = 138.05° ± 1.28°, ? = 271.15° ± 3.70°, ? = 315.26 ± 0.86°, and ? = 228.12°. We investigated whether this meteoroid stream could have originated from comets C/1978 T3 (Bradfield), C/1808 F1 (Pons), or C/1939 H1 (Jurlof-Achmarof-Hassel). If the parent body can be identified, we can determine when the comet was first captured into a low perihelion distance orbit. Future examination of the shower will allow us to examine the physical properties of the parent comet.

  14. Best Management Practices for Equine Disease Prevention 

    E-print Network

    Scott, Brett D.

    2008-10-06

    ;however,theseproductshave beenusedbysomeownersbecauseofthehighmortality rateinfoalswithclostridialenterocolitis.Adversereac- tionstothe Clostridium perfringens typeCandDtoxoid havebeenreportedinbroodmares. ContagiousEquineMetritis?Anacute,highlyconta- giousvenerealdiseaseofhorses...

  15. Simultaneous and enhanced production of thermostable amylases and ethanol from starch by cocultures of Clostridium thermosulfurogenes and Clostridium thermohydrosulfuricum

    SciTech Connect

    Hyun, H.H.; Zeikus, J.G.

    1985-05-01

    Clostridium thermohydrosulfuricum and Clostridium thermosulfurogenes produced ethanol and amylases with different components as primary metabolites of starch fermentation. Starch fermentation parameters were compared in mono- and cocultures of these two thermoanaerobes to show that the fermentation was dramatically improved as a consequence of coordinate action of amylolytic enzymes and synergistic metabolic interactions between the two species. Under given monoculture fermentation conditions, neither species completely degraded starch during the time course of the study, whereas in coculture, starch was completely degraded. In monoculture starch fermentation, C. thermohydrosulfuricum produced lower levels of pullulanase and glucoamylase, whereas C. thermosulfurogenes produced lower levels of ..beta..-amylase and glucoamylase. In coculture fermentation, improvement of starch metabolism by each species was noted in terms of increased amounts and rates of increased starch consumption, amylase production, and ethanol formation. The single-step coculture fermentation completely degraded 2.5% starch in 30 h at 60/sup 0/C and produced 9 U of ..beta..-amylase per ml, 1.3 U of pullulanase per ml, 0.3 U of glucoamylase per ml, and > 120 mM ethanol with a yield of 1.7 mol/mol of glucose in starch. The potential industrial applications of the coculture fermentation and the physiological basis for the interspecies metabolic interactions are discussed.

  16. Bacterial cellulose hydrolysis in anaerobic environmental subsystems--Clostridium thermocellum and Clostridium stercorarium, thermophilic plant-fiber degraders.

    PubMed

    Zverlov, Vladimir V; Schwarz, Wolfgang H

    2008-03-01

    Cellulose degradation is a rare trait in bacteria. However, the truly cellulolytic bacteria are extremely efficient hydrolyzers of plant cell wall polysaccharides, especially those in thermophilic anaerobic ecosystems. Clostridium stercorarium, a thermophilic ubiquitous soil dweller, has a simple cellulose hydrolyzing enzyme system of only two cellulases. However, it seems to be better suited for the hydrolysis of a wide range of hemicelluloses. Clostridium thermocellum, an ubiquitous thermophilic gram-type positive bacterium, is one of the most successful cellulose degraders known. Its extracellular enzyme complex, the cellulosome, was prepared from C. thermocellum cultures grown on cellulose, cellobiose, barley beta-1,3-1,4-glucan, or a mixture of xylan and cellulose. The single proteins were identified by peptide chromatography and MALDI-TOF-TOF. Eight cellulosomal proteins could be found in all eight preparations, 32 proteins occur in at least one preparation. A number of enzymatic components had not been identified previously. The proportion of components changes if C. thermocellum is grown on different substrates. Mutants of C. thermocellum, devoid of scaffoldin CipA, that now allow new types of experiments with in vitro cellulosome reassembly and a role in cellulose hydrolysis are described. The characteristics of these mutants provide strong evidence of the positive effect of complex (cellulosome) formation on hydrolysis of crystalline cellulose. PMID:18378600

  17. Purification and characterization of Clostridium sordellii hemorrhagic toxin and cross-reactivity with Clostridium difficile toxin A (enterotoxin).

    PubMed Central

    Martinez, R D; Wilkins, T D

    1988-01-01

    Hemorrhagic toxin (toxin HT) was purified from Clostridium sordellii culture filtrate. The purification steps included ultrafiltration through an XM-100 membrane filter and immunoaffinity chromatography, using a monoclonal antibody to toxin A of Clostridium difficile as the ligand. Toxin HT migrated as a major band with a molecular weight of 525,000 and a minor band at 450,000 on nondenaturing gradient polyacrylamide gel electrophoresis. The molecular weight was estimated at 300,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Isoelectric focusing indicated an apparent pI of 6.1. Toxin HT was cytotoxic for cultured cells and lethal for mice by intraperitoneal injection, and it elicited an accumulation of hemorrhagic fluid in rabbit ileal loops. Immunodiffusion analysis revealed a reaction of partial identity between toxins A and HT. Immunological cross-reactivity between these toxins was further demonstrated by immunoblotting and by neutralization of toxin HT biological activity with antibodies to toxin A. A sensitive indirect enzyme-linked immunosorbent assay was used to examine the affinity involved in homologous and heterologous antigen-antibody interactions. Our findings show that toxin HT has biological activities and immunological properties similar to those of toxin A; however, the toxins are not identical. Images PMID:3128481

  18. Clostridium paraputrificum Bacteremia Associated with Colonic Necrosis in a Patient with AIDS

    PubMed Central

    Shinha, Takashi; Hadi, Christiane

    2015-01-01

    Clostridium species are anaerobic Gram-positive rods that can cause a broad range of invasive infections in humans, including myonecrosis and bacteremia. Importantly, clostridial bacteremia is frequently associated with underlying medical conditions, such as colonic malignancy. Characterizing Clostridium spp. and understanding their associated clinical disease spectrum are paramount to provide optimal treatment, thereby decreasing morbidity and mortality especially in those with underlying debilitating comorbidities. Clostridium paraputrificum is an infrequently isolated Clostridium species and its clinical significance has not been well described. We herein report a case of bacteremia due to C. paraputrificum in a 65-year-old man with AIDS who developed acute colonic necrosis complicated by septic shock. We then review other cases of bacteremia associated with C. paraputrificum in the literature in addition to discussing the clinical significance of anaerobic bacteremia in general. To our knowledge, our report is the second case of C. paraputrificum bacteremia in a patient with AIDS. PMID:25692054

  19. Calculations of Diffuser Flows with an Anisotropic K-Epsilon Model

    NASA Technical Reports Server (NTRS)

    Zhu, J.; Shih, T.-H.

    1995-01-01

    A newly developed anisotropic K-epsilon model is applied to calculate three axisymmetric diffuser flows with or without separation. The new model uses a quadratic stress-strain relation and satisfies the realizability conditions, i.e., it ensures both the positivity of the turbulent normal stresses and the Schwarz' inequality between any fluctuating velocities. Calculations are carried out with a finite-element method. A second-order accurate, bounded convection scheme and sufficiently fine grids are used to ensure numerical credibility of the solutions. The standard K-epsilon model is also used in order to highlight the performance of the new model. Comparison with the experimental data shows that the anisotropic K-epsilon model performs consistently better than does the standard K-epsilon model in all of the three test cases.

  20. Low Reynolds number k-epsilon modelling with the aid of direct simulation data

    NASA Technical Reports Server (NTRS)

    Rodi, W.; Mansour, N. N.

    1993-01-01

    The constant C sub mu and the near-wall damping function f sub mu in the eddy-viscosity relation of the k-epsilon model are evaluated from direct numerical simulation (DNS) data for developed channel and boundary layer flow at two Reynolds numbers each. Various existing f sub mu model functions are compared with the DNS data, and a new function is fitted to the high-Reynolds-number channel flow data. The epsilon-budget is computed for the fully developed channel flow. The relative magnitude of the terms in the epsilon-equation is analyzed with the aid of scaling arguments, and the parameter governing this magnitude is established. Models for the sum of all source and sink terms in the epsilon-equation are tested against the DNS data, and an improved model is proposed.

  1. Degeneration of solventogenic Clostridium strains monitored by Fourier transform infrared spectroscopy of bacterial cells

    Microsoft Academic Search

    KC Schuster; R Goodacre; JR Gapes; M Young

    2001-01-01

      Strain degeneration in solventogenic clostridia is a known problem in the technical acetone–butanol fermentation bioprocess,\\u000a especially in the continuous process mode. Clostridial strain degeneration was studied by Fourier transform infrared (FT-IR)\\u000a spectroscopy of the bacterial cells. Degenerative variant formation in two strains, Clostridium beijerinckii NCIMB 8052 and Clostridium species AA332, was detected spectroscopically. Colonies on solid media were sampled, or

  2. Attenuation of epsilon(sub eff) of coplanar waveguide transmission lines on silicon substrates

    NASA Technical Reports Server (NTRS)

    Taub, Susan R.; Young, Paul G.

    1993-01-01

    Attenuation and epsilon(sub eff) of Coplanar Waveguide (CPW) transmission lines were measured on Silicon substrates with resistivities ranging from 400 to greater than 30,000 ohm-cm, that have a 1000 angstrom coating of SiO2. Both attenuation and epsilon(sub eff) are given over the frequency range 5 to 40 GHz for various strip and slot widths. These measured values are also compared to the theoretical values.

  3. Comparison of differential Reynolds stress and k-epsilon turbulence models for the driven cavity problem

    Microsoft Academic Search

    R. K. Cooper; M. Wolfshtein; M. Behnia; G. de Vahl Davis; J. Reizes

    1988-01-01

    The differential Reynolds stress (RS) transport model for turbulent flow is used to study recirculating flow (driven by a moving wall) in a rectangular cavity. Empirical cavity jet boundary conditions based on wall-jet and cavity-jet data are obtained. Good mean velocity field predictions are obtained with both the k-epsilon and RS-epsilon models. The stabilizing effect of flow curvature is well

  4. Scrutinizing the k-epsilon turbulence model under adverse pressure gradient conditions

    Microsoft Academic Search

    W. Rodi; G. Scheuerer

    1986-01-01

    The k-epsilon model and a one-equation model have been used to predict adverse pressure gradient boundary layers. While the one-equation model gives generally good results, the k-epsilon model reveals systematic discrepancies, e.g., excessively high skin friction coefficients, for these relatively simple flows. These shortcomings are examined and it is shown by an analytical analysis for the log-law region that the

  5. On the vortex stretching modification of the k-epsilon turbulence model - Radial jets

    Microsoft Academic Search

    A. Rubel

    1985-01-01

    The invariant vortex stretching modification to the kappa-epsilon model is applied to the self-preserving radial jet and shown to be inadequate. Results are obtained which support the contention that the Pope (1978) modification to the kappa-epsilon model affects a radial jet more substantially than a round one. It is also noted that radial far-field behavior can impose an additional constraint

  6. Scrutinizing the k-epsilon-model under adverse pressure gradient conditions

    Microsoft Academic Search

    W. Rodi; G. Scheuerer

    1984-01-01

    The k-epsilon model and a one-equation model have been used to predict adverse pressure gradient boundary layers. While the one-equation model gives generally good results, the k-epsilon model reveals systematic discrepancies, e.g., too high skin friction coefficients for these relatively simple flows. These shortcomings are examined and it is shown by an analytical analysis for the log-law region that the

  7. Flow over a cylinder at a plane boundary - A model based upon (k - epsilon) turbulence

    Microsoft Academic Search

    T. Solberg; K. J. Eidsvik

    1989-01-01

    A model for two-dimensional flows over a cylinder at a plane boundary is developed. The model, based upon a (k-epsilon) turbulence closure, is formulated in a curvilinear coordinate system based upon frictionless flow. A length scale modification in areas of adverse pressure gradient and recirculating flow appears to be more realistic than the standard (k-epsilon) model. The main features of

  8. A multi-zone k-epsilon turbulence model for complex configurations

    Microsoft Academic Search

    Beth W. Lowrie

    1990-01-01

    The Jones-Launder (1973) low-Reynolds-number form of the k-epsilon equations has been extended to provide the capability for computing flows for complex geometries. This multizone k-epsilon model allows computation of geometries which otherwise could not be computed due to grid-generation constraints or memory limitations. This extension required generalization of the low-Reynolds-number terms and development of a multizone characteristic boundary condition consistent

  9. A new low-Reynolds-number k-epsilon-fmu model for predictions involving multiple surfaces

    Microsoft Academic Search

    Tae Seon Park; Hyung Jin Sung

    1997-01-01

    A new k-epsilon-fmu turbulence model is proposed, in which the near-wall effect without reference to distance and the non-equilibrium effect are incorporated. In this model, the non-local near-wall effect in a general coordinate system is taken into account by the fmu equation, and the local anisotropy in strongly strained turbulent flows in introduced in the epsilon equation. The near-wall characteristics

  10. Comparative study of high and low Reynolds number versions of k-epsilon models

    Microsoft Academic Search

    Ram K. Avva; Clifford E. Smith; Ashok K. Singhal

    1990-01-01

    Two commonly used turbulence models for predicting wall-bounded flows are: (1) the high Reynolds number k-epsilon model with wall functions and (2) the low Reynolds number k-epsilon model with near-wall resolution, often claimed to be more accurate than wall functions. The present study is aimed at the systematic assessment of the two approaches by considering a number of two-dimensional flows.

  11. Wall functions for the kappa-epsilon turbulence model in generalized nonorthogonal curvilinear coordinates

    Microsoft Academic Search

    D. L. Sondak; R. H. Pletcher; W. R. Vandalsem

    1992-01-01

    A k-epsilon turbulence model suitable for compressible flow, including the new wall function formulation, has been incorporated into an existing compressible Reynolds-averaged Navier-Stokes code, F3D. The low Reynolds number k-epsilon model of Chien (1982) was added for comparison with the present method. A number of features were added to the F3D code including improved far-field boundary conditions and viscous terms

  12. K-epsilon model of anomalous transport in resistive interchange turbulence

    Microsoft Academic Search

    H. Sugama; M. Okamoto; M. Wakatani

    1992-01-01

    A K(sub epsilon) anomalous transport model for resistive interchange turbulence is presented and applied to the transport analysis of ECH plasmas in Heliotron E. In this model, the turbulent kinetic energy K approximately equals 1\\/2(upsilon tilde(sup 2)) and its viscous dissipation rate epsilon characterize the local turbulence and the anomalous transport coefficient is given by D approximately equal to K(sup

  13. Wall functions for the kappa-epsilon turbulence model in generalized nonorthogonal curvilinear coordinates. Final Report

    Microsoft Academic Search

    D. L. Sondak; R. H. Pletcher; W. R. Vandalsem

    1992-01-01

    A k-epsilon turbulence model suitable for compressible flow, including the new wall function formulation, has been incorporated into an existing compressible Reynolds-averaged Navier-Stokes code, F3D. The low Reynolds number k-epsilon model of Chien (1982) was added for comparison with the present method. A number of features were added to the F3D code including improved far-field boundary conditions and viscous terms

  14. A new epsilon globin HincII variant fragment length in a South African Negroid family

    Microsoft Academic Search

    M Ramsay; J A Thomson; T Jenkins

    1986-01-01

    A new HincII epsilon globin variant is reported in a South African Negroid family. The usual HincII epsilon globin fragment lengths are 8.0 and 3.7 kb and the variant described here is 14.0 kb in length. The 14.0 kb fragment was generated by a site alteration removing the 3' HincII site on a chromosome that already lacked the 5' HincII

  15. Phase stability of {epsilon} and {gamma} HNIW (CL-20) at high-pressure and temperature

    SciTech Connect

    Gump, Jared C.; Stoltz, Chad A.; Peiris, Suhithi M. [Indian Head Division-Naval Surface Warfare Center, Indian Head, MD 20640 (United States)

    2007-12-12

    Hexanitrohexaazaisowurtzitane (CL-20) is one of the few ingredients developed since World War II to be considered for transition to military use. Five polymorphs have been identified for CL-20 by FTIR measurements ({alpha}, {beta}, {gamma}, {epsilon}, {zeta}). As CL-20 is transitioned into munitions it will become necessary to predict its response under conditions of detonation, for performance evaluation. Such predictive modeling requires a phase diagram and basic thermodynamic properties of the various phases at high pressure and temperature. Therefore, the epsilon and gamma phases of CL-20 at static high-pressure and temperature were investigated using synchrotron angle-dispersive x-ray diffraction experiments. The samples were compressed and heated using diamond anvil cells (DAC). Pressures and temperatures achieved were around 5 GPa and 240 deg. C, respectively. The epsilon phase was stable to 6.3 GPa at ambient temperature. When heated at ambient pressure the epsilon phase was sustained to a temperature of 120 deg. C then underwent a transition to the gamma phase above 125 deg. C and then thermal decomposition occurred above 150 deg. C. Upon compression, the gamma phase underwent a phase transition at both ambient temperature and 140 deg. C. Pressure--volume data for the epsilon and gamma phase at ambient temperature and the epsilon phase at 75 deg. C were fit to the Birch-Murnaghan formalism to obtain isothermal equations of state.

  16. Deep L' and M-band Imaging for Planets Around Vega and epsilon Eridani

    E-print Network

    A. N. Heinze; Philip M. Hinz; Matthew Kenworthy; Douglas Miller; Suresh Sivanandam

    2008-07-24

    We have obtained deep Adaptive Optics (AO) images of Vega and epsilon Eri to search for planetary-mass companions. We observed at the MMT in the L' (3.8 micron) and M (4.8 micron) bands using Clio, a recently commissioned imager optimized for these wavelengths. Observing at these long wavelengths represents a departure from the H band (1.65 microns) more commonly used for AO imaging searches for extrasolar planets. The long wavelengths offer better predicted planet/star flux ratios and cleaner (higher Strehl) AO images, at the cost of lower diffraction limited resolution and higher sky background. We have not detected any planets or planet candidates around Vega or epsilon Eri. We report the sensitivities obtained around both stars, which correspond to upper limits on any planetary companions which may exist. The sensitivities of our L' and M band observations are comparable to those of the best H-regime observations of these stars. For epsilon Eri our M band observations deliver considerably better sensitivity to close-in planets than any previously published results, and we show that the M band is by far the best wavelength choice for attempts at ground-based AO imaging of the known planet epsilon Eri b. The Clio camera itself with MMTAO may be capable of detecting epsilon Eri b at its 2010 apastron, given a multi-night observing campaign. Clio appears to be the only currently existing AO imager that has a realistic possibility of detecting epsilon Eri b.

  17. Clostridium difficile in a children's hospital: assessment of environmental contamination.

    PubMed

    Warrack, Simone; Duster, Megan; Van Hoof, Sarah; Schmitz, Michelle; Safdar, Nasia

    2014-07-01

    Clostridium difficile infection (CDI) is the most frequent infectious cause of health care-associated diarrhea. Three cases of CDI, in children age 2, 3, and 14 years, occurred in the hematology/oncology ward of our children's hospital over 48 hours. We aimed to assess environmental contamination with C difficile in the shared areas of this unit, and to determine whether person-to-person transmission occurred. C difficile was recovered from 5 of 18 samples (28%). We compared C difficile isolated from each patient and the environment using pulsed-field gel electrophoresis, and found that none of the patient strains matched any of the others, and that none matched any strains recovered from the environment, suggesting that person-to-person transmission had not occurred. We found that C difficile was prevalent in the environment throughout shared areas of the children's hospital unit. Molecular typing to identify mechanisms of transmission is useful for devising appropriate interventions. PMID:24751141

  18. Organization and distribution of the cellulosome in Clostridium thermocellum.

    PubMed Central

    Bayer, E A; Setter, E; Lamed, R

    1985-01-01

    The properties of the cellulosome (the cellulose-binding, multicellulase-containing protein complex) in Clostridium thermocellum were examined by comparing the cellulase systems derived from the wild type and an adherence-defective mutant. The growth conditions--specifically, growth either on cellulose (Avicel) or on cellobiose as insoluble or soluble carbon sources, respectively--were found to be critical to the distribution of the cellulosome in the mutant system: the cellobiose-grown mutant (in contrast to the wild type) lacked the cellulosome on its surface and produced only minor quantities of the extracellular cellulosome accompanied by other relatively low-molecular-weight cellulases. The polypeptide composition of the respective purified cellulosome was dependent on the nature of the carbon source and was similar for both wild-type and mutant cells. Ultrastructural analysis revealed the presence of novel polycellulosomal protuberances on the cell surface of the cellobiose-grown wild type which were absent in the mutant. Images PMID:4019409

  19. Integration of metabolism and virulence in Clostridium difficile.

    PubMed

    Bouillaut, Laurent; Dubois, Thomas; Sonenshein, Abraham L; Dupuy, Bruno

    2015-05-01

    Synthesis of the major toxin proteins of the diarrheal pathogen, Clostridium difficile, is dependent on the activity of TcdR, an initiation (sigma) factor of RNA polymerase. The synthesis of TcdR and the activation of toxin gene expression are responsive to multiple components in the bacterium's nutritional environment, such as the presence of certain sugars, amino acids, and fatty acids. This review summarizes current knowledge about the mechanisms responsible for repression of toxin synthesis when glucose or branched-chain amino acids or proline are in excess and the pathways that lead to synthesis of butyrate, an activator of toxin synthesis. The regulatory proteins implicated in these mechanisms also play key roles in modulating bacterial metabolic pathways, suggesting that C. difficile pathogenesis is intimately connected to the bacterium's metabolic state. PMID:25445566

  20. Current Treatment Options for Severe Clostridium difficile–associated Disease

    PubMed Central

    Surawicz, Christina M.

    2008-01-01

    A notable trend toward severe Clostridium difficile colitis and poor outcomes has emerged since recognition of the hyper-virulent C. difficile NAP1/027 strain. This trend has increased the emphasis on appropriate treatment regimens in refractory cases of C. difficile infection. In mild-to-moderate cases, oral metronidazole remains adequate first-line therapy, but in the absence of a good clinical response, switching to vancomycin may be necessary. Oral vancomycin should be used as initial therapy in severely ill patients or patients who cannot tolerate metronidazole. Rectal administration of vancomycin may be used as adjunctive therapy for severely ill patients. Patients with an ileus who cannot tolerate oral medications may improve with adjunct intravenous metronidazole and/or rectal vancomycin. Early surgical consultation should be requested, as some patients will require emergent colectomy. The shifting landscape of C. difficile infection has undermined our complacency regarding this long-recognized disease. PMID:21904490

  1. Enzyme-linked immunosorbent assay for Clostridium difficile toxin A.

    PubMed Central

    Lyerly, D M; Sullivan, N M; Wilkins, T D

    1983-01-01

    Antibodies against Clostridium difficile toxin A were purified by affinity chromatography from antiserum prepared against crude C. difficile toxin preparations. The affinity-purified antibody preparation was free of detectable amounts of antibodies to other C. difficile antigens, as demonstrated by crossed immunoelectrophoresis, and specifically neutralized the cytotoxicity of toxin A. An indirect enzyme-linked immunosorbent assay (ELISA) was subsequently developed using the antibody preparation for the specific detection of toxin A. The ELISA, which could detect 1 ng (5 ng/ml) of toxin A, was used to quantitate the toxin in the culture supernatant fluids of strains of C. difficile. The ELISA values for toxin A closely correlated with the toxin A and B cytotoxic titers of the supernatant fluids. In addition, toxin A was detected by ELISA in human fecal specimens from persons with antibiotic-associated colitis, demonstrating that this toxin is produced during C. difficile colitis. Images PMID:6338036

  2. Detection of Clostridium difficile toxins by enzyme immunoassay.

    PubMed Central

    Krishnan, C.

    1986-01-01

    An enzyme-linked immunosorbent assay (ELISA) for the rapid diagnosis of antibiotic-associated colitis (AAC) is presented. Commercially available antisera to Clostridium difficile toxins contain antibodies to other antigens found in non-toxigenic C. difficile and other bacteria. Removal of these unwanted antibodies by absorption increased the specificity of ELISA for detection of C. difficile toxins. Specimens tested included 40 faecal extracts positive for cytotoxicity from cases of AAC, 30 diarrhoeic and 30 well-formed stools negative for cytotoxicity and 50 culture filtrates of toxigenic and non-toxigenic C. difficile and other clostridial species. Use of absorbed sera reduced false-positive reactions observed with faecal specimens from 23 to 8%. About 90% of specimens that were positive by the tissue culture cytotoxicity test were positive by ELISA using the absorbed sera. The relative merits of ELISA and other methods for the rapid diagnosis of AAC are discussed. PMID:3950397

  3. Clostridium difficile: improving the prevention paradigm in healthcare settings.

    PubMed

    Vassallo, Angela; Tran, Mai-Chi N; Goldstein, Ellie J C

    2014-09-01

    Clostridium difficile infection (CDI) is a major public health problem worldwide with significant morbidity and mortality that is spread by spores and fecal oral transmission. A variety of risk factors have been identified. Some risk factors such as age, are not amenable to change, while others such as antimicrobial utilization have resulted in broadly implemented antimicrobial stewardship programs. New risk factors are emerging such as proton pump inhibitor (PPI) use, irritable bowel disease (IBD) and obesity, with others yet to be determined. Prevention of spread of CDI is imperative, since therapy remains imperfect. We review established and emerging risks for CDI and offer potential preventative strategies with the use of a multidisciplinary CDI prevention bundle checklist. PMID:25109301

  4. Clostridium difficile infection in the twenty-first century

    PubMed Central

    Ghose, Chandrabali

    2013-01-01

    Clostridium difficile is a spore-forming gram-positive bacillus, and the leading cause of antibiotic-associated nosocomial diarrhea and colitis in the industrialized world. With the emergence of a hypervirulent strain of C. difficile (BI/NAP1/027), the epidemiology of C. difficile infection has rapidly changed in the last decade. C. difficile infection, once thought to be an easy to treat bacterial infection, has evolved into an epidemic that is associated with a high rate of mortality, causing disease in patients thought to be low-risk. In this review, we discuss the changing face of C .difficile infection and the novel treatment and prevention strategies needed to halt this ever growing epidemic. PMID:26038491

  5. Clostridium difficile infection after ileostomy closure mimicking anastomotic leak.

    PubMed

    Almerie, Muhammad Qutayba; Culverwell, Adam; Mahon, Christopher

    2015-01-01

    Clostridium difficile infection is linked to antibiotic exposure, with elderly and immunocompromised hospitalised patients being particularly at risk. The symptoms range from mild diarrhoea to life-threatening fulminant colitis. We describe an unusual presentation of C. difficile infection after closure of ileostomy in a healthy 60-year-old man with a history of low anterior resection and defunctioning ileostomy for rectal tumour. On the third day postoperatively, the patient developed left lower abdominal pain and profuse diarrhoea. With worsening symptoms and steadily increasing inflammatory markers over the following few days, concerns were raised about an anastomotic leak with pelvic abscess. CT of the abdomen/pelvis on day 7 surprisingly showed colitis in the neorectum/sigmoid colon. A stool test confirmed C. difficile infection. PMID:26150639

  6. Recurrent Clostridium difficile infection: From colonization to cure.

    PubMed

    Shields, Kelsey; Araujo-Castillo, Roger V; Theethira, Thimmaiah G; Alonso, Carolyn D; Kelly, Ciaran P

    2015-08-01

    Clostridium difficile infection (CDI) is increasingly prevalent, dangerous and challenging to prevent and manage. Despite intense national and international attention the incidence of primary and of recurrent CDI (PCDI and RCDI, respectively) have risen rapidly throughout the past decade. Of major concern is the increase in cases of RCDI resulting in substantial morbidity, morality and economic burden. RCDI management remains challenging as there is no uniformly effective therapy, no firm consensus on optimal treatment, and reliable data regarding RCDI-specific treatment options is scant. Novel therapeutic strategies are critically needed to rapidly, accurately, and effectively identify and treat patients with, or at-risk for, RCDI. In this review we consider the factors implicated in the epidemiology, pathogenesis and clinical presentation of RCDI, evaluate current management options for RCDI and explore novel and emerging therapies. PMID:25930686

  7. Immunization Using GroEL Decreases Clostridium difficile Intestinal Colonization

    PubMed Central

    Péchiné, Séverine; Hennequin, Claire; Boursier, Céline; Hoys, Sandra; Collignon, Anne

    2013-01-01

    Clostridium difficile is a pathogen which is responsible for diarrhea and colitis, particularly after treatment with antibiotics. Clinical signs are mainly due to two toxins, TcdA and TcdB. However, the first step of pathogenesis is the colonization process. We evaluated C. difficile surface proteins as vaccine antigens in the hamster model to prevent intestinal colonization. This vaccination induced a partial protection of hamsters against death after a C. difficile challenge. A proteomic analysis of animal sera allowed us to identify proteins which could be responsible for the protection observed. Among these proteins, we identified the GroEL heat shock protein. To confirm the role of the specific GroEL antibodies in the delayed C. difficile colonization of hamsters, we performed an immunization assay in a mouse model. After intranasal immunization with the recombinant protein GroEL, we observed a lower C. difficile intestinal colonization in the immunized group as compared to the control group. PMID:24303034

  8. Probiotics in Antibiotic-Associated Diarrhea and Clostridium difficile Infection

    Microsoft Academic Search

    Hania Szajewska

    \\u000a \\u000a \\u000a \\u000a \\u000a \\u000a • \\u000a \\u000a \\u000a Antibiotic-associated diarrhea (AAD) is a common pediatric disorder.\\u000a \\u000a \\u000a \\u000a • \\u000a \\u000a \\u000a This chapter summarizes the available evidence on the efficacy of probiotics in children in the prevention and treatment of\\u000a any diarrhea associated with the use of antibiotics and that was caused by Clostridium difficile .\\u000a \\u000a \\u000a \\u000a \\u000a • \\u000a \\u000a \\u000a Results emerging from pediatric trials provide evidence of a moderate beneficial effect of selected

  9. Continuous xylose fermentation by Clostridium acetobutylicum - Assessment of solventogenic kinetics.

    PubMed

    Procentese, Alessandra; Raganati, Francesca; Olivieri, Giuseppe; Russo, Maria Elena; Salatino, Piero; Marzocchella, Antonio

    2015-09-01

    This work deals with the specific butanol production rate of Clostridium acetobutylicum using xylose - a relevant fraction of lignocellulosic feedstock for biofuel production - as carbon source. The tests were carried out in a CSTR equipped with a microfiltration unit. The dilution rate (D) ranged between 0.02 and 0.22h(-1) and the ratio R between the permeate stream rate and the stream fed to the reactor ranged between 14% and 88%. The biomass present in the broth was identified as a heterogeneous cell population consisting of: acidogenic cells, solventogenic cells and spores. The results were processed to assess the concentration of acidogenic cells, solventogenic cells and spores. The specific butanol production rate was also assessed. The max butanol productivity was 1.3gL(-1)h(-1) at D=0.17h(-1) and R=30%. A comparison between the results reported in a previous work carried out with lactose was made. PMID:26025352

  10. Thermal sensitivity of Clostridium botulinum type C toxin.

    PubMed Central

    Hubálek, Z.; Halouzka, J.

    1988-01-01

    A sterile suspension containing 950 mouse LD50 per ml of type C botulinum toxin was exposed for various periods to different temperatures. The time required for the 99% (hundred-fold) reduction of toxicity was more than 5 years at -70 degrees C or -20 degrees C, 6 months at +5 degrees C, 3 weeks at +20 degrees C, 2 weeks at +28 degrees C, 2 days at +37 degrees C, 9 h at +42 degrees C, less than 30 min at +56 degrees C, less than 20 min at +60 degrees C, and below 5 min at +80 degrees C. The results suggest that Clostridium botulinum type C toxin, if produced in an ecosystem of the mild climatic zone, might persist there over the winter season and cause the intoxication of vertebrates next early spring in the absence of further microbial toxigenesis. PMID:2972554

  11. Ipilimumab-associated colitis or refractory Clostridium difficile infection?

    PubMed

    Gupta, Arjun; Khanna, Sahil

    2015-01-01

    We present a case of a patient with a diagnostic dilemma who was referred for possible faecal microbiota transplantation (FMT) for refractory diarrhoea secondary to Clostridium difficile infection (CDI). On detailed history, the patient was exposed to ipilimumab concomitantly while being treated for CDI, and was instead diagnosed with diarrhoea secondary to superimposed ipilimumab-associated colitis. Ipilimumab is an anti-CTLA4 monoclonal antibody approved for use in metastatic melanoma and under trial for other indications. Ipilimumab is associated with several immune-related adverse effects, of which diarrhoea and colitis are the most common. While FMT has shown tremendous efficacy in managing recurrent and refractory CDI, it was not offered in this case due to negative C. difficile testing showing a high degree of suspicion for ipilimumab-associated colitis due to recent drug use. Our patient was successfully managed with fluid resuscitation and steroids, and remains symptom free at last follow-up at 9?months. PMID:26153295

  12. Hydrogen production by immobilized whole cells of Clostridium butyricum

    NASA Astrophysics Data System (ADS)

    Suzuki, S.; Karube, I.

    Immobilized microbial cells were used in a batch system in an attempt to achieve continuous hydrogen production from glucose and waste waters. Clostridium butyricum IFO 3847 was immobilized in polyacrylamide gel and continuously produced hydrogen from glucose. The hydrogen producing bacteria were then immobilized in 2% agar gel and showed continuous hydrogen production from an alcohol factory's waste waters. The hydrogen production rate became constant above BOD 1500 ppm when performed with a batch system. The immobilized whole cells continuously produced hydrogen over a 20 day period, producing about 6 ml/min/kg wet gels. Hydrogen production by bacteria immobilized in acetylcellulose filters was six times higher than that by cells entrapped in agar gels.

  13. Bile Salts and Glycine as Cogerminants for Clostridium difficile Spores?

    PubMed Central

    Sorg, Joseph A.; Sonenshein, Abraham L.

    2008-01-01

    Spore formation by Clostridium difficile is a significant obstacle to overcoming hospital-acquired C. difficile-associated disease. Spores are resistant to heat, radiation, chemicals, and antibiotics, making a contaminated environment difficult to clean. To cause disease, however, spores must germinate and grow out as vegetative cells. The germination of C. difficile spores has not been examined in detail. In an effort to understand the germination of C. difficile spores, we characterized the response of C. difficile spores to bile. We found that cholate derivatives and the amino acid glycine act as cogerminants. Deoxycholate, a metabolite of cholate produced by the normal intestinal flora, also induced germination of C. difficile spores but prevented the growth of vegetative C. difficile. A model of resistance to C. difficile colonization mediated by the normal bacterial flora is proposed. PMID:18245298

  14. Clostridium difficile-associated reactive arthritis in two children.

    PubMed

    Löffler, Helga A; Pron, Benedicte; Mouy, Richard; Wulffraat, Nico M; Prieur, Anne-Marie

    2004-01-01

    In adults, reactive arthritis (ReA) following Clostridium difficile-enterocolitis has been documented. In children, only one case of C. difficile-associated ReA has been reported. We now describe two other cases of ReA associated with C. difficile in children. The characteristics of ReA due to C. difficile appear to be similar in adults and children. Both children show polyarthritis after an episode of diarrhoea with positive stool cultures for C. difficile. Arthritis is asymmetrical with a self-limiting course. Nonsteroidal antiinflammatory drug (NSAID) therapy is sufficient. One case is remarkable because of its prolonged course of ReA despite NSAID therapy, and its association with the presence of HLA-B27 antigen. PMID:14769523

  15. 21 CFR 184.1983 - Bakers yeast extract.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...than 10,000 organisms/gram by aerobic plate count. (2) Less than 10 yeasts and molds/gram. (3) Negative for Salmonella, E. coli, coagulase positive Staphylococci, Clostridium perfringens, Clostridium botulinum, or any other...

  16. Rle du lactose dans la formation d'entrocolite ulcroncrosante (ecun) exprimentale chez la caille gnotoxnique monoassocie des souches de Clostridium butyricum

    E-print Network

    Boyer, Edmond

    'hydrogène. Howard F. M. et al., 1977. Outbreak of necrotizing enterocolitis caused by Clostridium butyricum. Lancei-associated with Clostridium butyricum strains isolated from patients with necrotizing enterocolitis. lnfect. lmmun., 47, 697

  17. Deuterium nuclear magnetic resonance studies on the plasmalogens and the glycerol acetals of plasmalogens of Clostridium butyricum and Clostridium beijerinckii

    SciTech Connect

    Malthaner, M.; Seelig, J.; Johnston, N.C.; Goldfine, H.

    1987-09-08

    Deuterium nuclear magnetic resonance was used to investigate the structure of different lipid fractions isolated from the anaerobic bacteria Clostridium butyricum and Clostridium beijerinckii. The fractions isolated from C. butyricum were (1) phosphatidylethanolamine/plasmenylethanolamine and (2) the glycerol acetal of plasmenylethanolamine, and from C. beijerinckii similar fractions containing principally (1) phosphatidyl-N-monomethylethanolamine, along with its plasmalogen, and (2) the glycerol acetal of this plasmalogen were isolated. The third fraction from both species consisted largely of the acidic lipids phosphatidylglycerol and cardiolipin along with plasmalogen forms of these lipids. Palmitic acid with deuterium labels at C-2, C-3, or C-4 or oleic acid with deuterium labels at C-2 and C-9,10 was added to the growth medium and incorporated to various extents in the lipid fractions. Biochemical analysis showed that palmitic acid and oleic acid were preferentially bound to the sn-2 and sn-1 positions, respectively, of the glycerol backbone when both fatty acids were added to the medium. From the /sup 2/H NMR spectra, the hydrocarbon chain ordering near the lipid-water interface could be determined and appeared to be similar for all three lipid fractions. The deuterium quadrupole splitting and order parameter were low at the C-2 segment and increased by almost a factor of 2 at positions C-3 and C-4 for cells fed with deuteriated palmitic acid along with unlabeled oleic acid. These results agree with previous findings on pure diacyl lipids in which the sn-2 chain was found to adopt a bent conformation at the carbon segment C-2. However, two unusual quadrupole splittings could be detected for the plasmalogens. By comparison with other model systems it could be concluded that the double bond is aligned essentially parallel with the long axis of the hydrocarbon chains.

  18. Implications of Genome-Based Discrimination between Clostridium botulinum Group I and Clostridium sporogenes Strains for Bacterial Taxonomy.

    PubMed

    Weigand, Michael R; Pena-Gonzalez, Angela; Shirey, Timothy B; Broeker, Robin G; Ishaq, Maliha K; Konstantinidis, Konstantinos T; Raphael, Brian H

    2015-08-15

    Taxonomic classification of Clostridium botulinum is based on the production of botulinum neurotoxin (BoNT), while closely related, nontoxic organisms are classified as Clostridium sporogenes. However, this taxonomic organization does not accurately mirror phylogenetic relationships between these species. A phylogenetic reconstruction using 2,016 orthologous genes shared among strains of C. botulinum group I and C. sporogenes clearly separated these two species into discrete clades which showed ?93% average nucleotide identity (ANI) between them. Clustering of strains based on the presence of variable orthologs revealed 143 C. sporogenes clade-specific genetic signatures, a subset of which were further evaluated for their ability to correctly classify a panel of presumptive C. sporogenes strains by PCR. Genome sequencing of several C. sporogenes strains lacking these signatures confirmed that they clustered with C. botulinum strains in a core genome phylogenetic tree. Our analysis also identified C. botulinum strains that contained C. sporogenes clade-specific signatures and phylogenetically clustered with C. sporogenes strains. The genome sequences of two bont/B2-containing strains belonging to the C. sporogenes clade contained regions with similarity to a bont-bearing plasmid (pCLD), while two different strains belonging to the C. botulinum clade carried bont/B2 on the chromosome. These results indicate that bont/B2 was likely acquired by C. sporogenes strains through horizontal gene transfer. The genome-based classification of these species used to identify candidate genes for the development of rapid assays for molecular identification may be applicable to additional bacterial species that are challenging with respect to their classification. PMID:26048939

  19. Impact of Feed Supplementation with Antimicrobial Agents on Growth Performance of Broiler Chickens, Clostridium perfringens and Enterococcus Counts, and Antibiotic Resistance Phenotypes and Distribution of Antimicrobial Resistance Determinants in Escherichia coli Isolates?

    PubMed Central

    Diarra, Moussa S.; Silversides, Fred G.; Diarrassouba, Fatoumata; Pritchard, Jane; Masson, Luke; Brousseau, Roland; Bonnet, Claudie; Delaquis, Pascal; Bach, Susan; Skura, Brent J.; Topp, Edward

    2007-01-01

    The effects of feed supplementation with the approved antimicrobial agents bambermycin, penicillin, salinomycin, and bacitracin or a combination of salinomycin plus bacitracin were evaluated for the incidence and distribution of antibiotic resistance in 197 commensal Escherichia coli isolates from broiler chickens over 35 days. All isolates showed some degree of multiple antibiotic resistance. Resistance to tetracycline (68.5%), amoxicillin (61.4%), ceftiofur (51.3%), spectinomycin (47.2%), and sulfonamides (42%) was most frequent. The levels of resistance to streptomycin, chloramphenicol, and gentamicin were 33.5, 35.5, and 25.3%, respectively. The overall resistance levels decreased from day 7 to day 35 (P < 0.001). Comparing treatments, the levels of resistance to ceftiofur, spectinomycin, and gentamicin (except for resistance to bacitracin treatment) were significantly higher in isolates from chickens receiving feed supplemented with salinomycin than from the other feeds (P < 0.001). Using a DNA microarray analysis capable of detecting commonly found antimicrobial resistance genes, we characterized 104 tetracycline-resistant E. coli isolates from 7- to 28-day-old chickens fed different growth promoters. Results showed a decrease in the incidence of isolates harboring tet(B), blaTEM, sulI, and aadA and class 1 integron from days 7 to 35 (P < 0.01). Of the 84 tetracycline-ceftiofur-resistant E. coli isolates, 76 (90.5%) were positive for blaCMY-2. The proportions of isolates positive for sulI, aadA, and integron class 1 were significantly higher in salinomycin-treated chickens than in the control or other treatment groups (P < 0.05). These data demonstrate that multiantibiotic-resistant E. coli isolates can be found in broiler chickens regardless of the antimicrobial growth promoters used. However, the phenotype and the distribution of resistance determinants in E. coli can be modulated by feed supplementation with some of the antimicrobial agents used in broiler chicken production. PMID:17827305

  20. Analysis of the unexplored features of rrs (16S rDNA) of the Genus Clostridium

    PubMed Central

    2011-01-01

    Background Bacterial taxonomy and phylogeny based on rrs (16S rDNA) sequencing is being vigorously pursued. In fact, it has been stated that novel biological findings are driven by comparison and integration of massive data sets. In spite of a large reservoir of rrs sequencing data of 1,237,963 entries, this analysis invariably needs supplementation with other genes. The need is to divide the genetic variability within a taxa or genus at their rrs phylogenetic boundaries and to discover those fundamental features, which will enable the bacteria to naturally fall within them. Within the large bacterial community, Clostridium represents a large genus of around 110 species of significant biotechnological and medical importance. Certain Clostridium strains produce some of the deadliest toxins, which cause heavy economic losses. We have targeted this genus because of its high genetic diversity, which does not allow accurate typing with the available molecular methods. Results Seven hundred sixty five rrs sequences (> 1200 nucleotides, nts) belonging to 110 Clostridium species were analyzed. On the basis of 404 rrs sequences belonging to 15 Clostridium species, we have developed species specific: (i) phylogenetic framework, (ii) signatures (30 nts) and (iii) in silico restriction enzyme (14 Type II REs) digestion patterns. These tools allowed: (i) species level identification of 95 Clostridium sp. which are presently classified up to genus level, (ii) identification of 84 novel Clostridium spp. and (iii) potential reduction in the number of Clostridium species represented by small populations. Conclusions This integrated approach is quite sensitive and can be easily extended as a molecular tool for diagnostic and taxonomic identification of any microbe of importance to food industries and health services. Since rapid and correct identification allows quicker diagnosis and consequently treatment as well, it is likely to lead to reduction in economic losses and mortality rates. PMID:21223548