Sample records for clostridium perfringens epsilon

  1. Clostridium perfringens Epsilon Toxin Increases the Small Intestinal Permeability in Mice and Rats

    PubMed Central

    Goldstein, Jorge; Morris, Winston E.; Loidl, César Fabián; Tironi-Farinatti, Carla; McClane, Bruce A.; Uzal, Francisco A.; Fernandez Miyakawa, Mariano E.

    2009-01-01

    Epsilon toxin is a potent neurotoxin produced by Clostridium perfringens types B and D, an anaerobic bacterium that causes enterotoxaemia in ruminants. In the affected animal, it causes oedema of the lungs and brain by damaging the endothelial cells, inducing physiological and morphological changes. Although it is believed to compromise the intestinal barrier, thus entering the gut vasculature, little is known about the mechanism underlying this process. This study characterizes the effects of epsilon toxin on fluid transport and bioelectrical parameters in the small intestine of mice and rats. The enteropooling and the intestinal loop tests, together with the single-pass perfusion assay and in vitro and ex vivo analysis in Ussing's chamber, were all used in combination with histological and ultrastructural analysis of mice and rat small intestine, challenged with or without C. perfringens epsilon toxin. Luminal epsilon toxin induced a time and concentration dependent intestinal fluid accumulation and fall of the transepithelial resistance. Although no evident histological changes were observed, opening of the mucosa tight junction in combination with apoptotic changes in the lamina propria were seen with transmission electron microscopy. These results indicate that C. perfringens epsilon toxin alters the intestinal permeability, predominantly by opening the mucosa tight junction, increasing its permeability to macromolecules, and inducing further degenerative changes in the lamina propria of the bowel. PMID:19763257

  2. Brain lesions associated with clostridium perfringens type D epsilon toxin in a Holstein heifer calf.

    PubMed

    Mete, A; Garcia, J; Ortega, J; Lane, M; Scholes, S; Uzal, F A

    2013-09-01

    A 6-month-old dairy heifer calf with no premonitory signs was acutely down after the morning feeding and could not rise. On presentation, the heifer was in right lateral recumbency and moribund with opisthotonus and left hind limb paddling. Following euthanasia, gross examination of the brain revealed multifocal loss of gray-white matter distinction and extensive petechiae throughout the brainstem. On histopathological examination, there was striking white matter edema and marked perivascular proteinaceous edema surrounding many arterioles and venules (microangiopathy), mainly in the white matter of the internal capsule, thalamus, midbrain, cerebellum, and cerebellar peduncles. The perivascular neuropil was strongly positive for Alzheimer precursor protein A4. Clostridium perfringens epsilon toxin was detected in the intestinal contents. This is the first report of microangiopathy in postneonatal cattle associated with the detection of epsilon toxin in the intestinal contents. PMID:23381925

  3. Retinal microvascular damage and vasogenic edema produced by Clostridium perfringens type D epsilon toxin in rats.

    PubMed

    Finnie, John W; Manavis, Jim; Casson, Robert J; Chidlow, Glyn

    2014-04-16

    When the brain is exposed to large circulating levels of Clostridium perfringens type D epsilon toxin (EXT), microvascular damage with resulting severe, generalized, vasogenic edema seems to be principally responsible for the ensuing acute, and frequently fatal, neurologic disorder. However, although the blood-retinal barrier resembles in many respects the blood-brain barrier, retinal changes in livestock with acute epsilon intoxication have not, to the authors' knowledge, been previously reported. In rats given an acute dose of ETX, retinal microvascular endothelial injury led to widespread vasogenic edema as assessed immunohistochemically by marked plasma albumin extravasation. As laboratory rodents are a good model of the domestic livestock disease produced by ETX, it is probable that the latter sustain some visual deficit when exposed to large doses of this potent neurotoxin. PMID:24741023

  4. Clostridium perfringens epsilon toxin: the third most potent bacterial toxin known.

    PubMed

    Alves, Guilherme Guerra; Machado de Ávila, Ricardo Andrez; Chávez-Olórtegui, Carlos Delfin; Lobato, Francisco Carlos Faria

    2014-12-01

    Epsilon toxin (ETX) is produced by Clostridium perfringens type B and D strains and causes enterotoxemia, a highly lethal disease with major impacts on the farming of domestic ruminants, particularly sheep. ETX belongs to the aerolysin-like pore-forming toxin family. Although ETX has striking similarities to other toxins in this family, ETX is often more potent, with an LD50 of 100 ng/kg in mice. Due to this high potency, ETX is considered as a potential bioterrorism agent and has been classified as a category B biological agent by the Centers for Disease Control and Prevention (CDC) of the United States. The protoxin is converted to an active toxin through proteolytic cleavage performed by specific proteases. ETX is absorbed and acts locally in the intestines then subsequently binds to and causes lesions in other organs, including the kidneys, lungs and brain. The importance of this toxin for veterinary medicine and its possible use as a biological weapon have drawn the attention of researchers and have led to a large number of studies investigating ETX. The aim of the present work is to review the existing knowledge on ETX from C. perfringens type B and D. PMID:25234332

  5. Clostridium perfringens epsilon-toxin increases permeability of single perfused microvessels of rat mesentery.

    PubMed

    Adamson, R H; Ly, J C; Fernandez-Miyakawa, M; Ochi, S; Sakurai, J; Uzal, F; Curry, F E

    2005-08-01

    Epsilon-toxin, the primary virulence factor of Clostridium perfringens type D, causes mortality in livestock, particularly sheep and goats, in which it induces an often-fatal enterotoxemia. It is believed to compromise the intestinal barrier and then enter the gut vasculature, from which it is carried systemically, causing widespread vascular endothelial damage and edema. Here we used single perfused venular microvessels in rat mesentery, which enabled direct observation of permeability properties of the in situ vascular wall during exposure to toxin. We determined the hydraulic conductivity (L(p)) of microvessels as a measure of the response to epsilon-toxin. We found that microvessels were highly sensitive to toxin. At 10 microg ml(-1) the L(p) increased irreversibly to more than 15 times the control value by 10 min. At 0.3 microg ml(-1) no increase in L(p) was observed for up to 90 min. The toxin-induced increase in L(p) was consistent with changes in ultrastructure of microvessels exposed to the toxin. Those microvessels exhibited gaps either between or through endothelial cells where perfusate had direct access to the basement membrane. Many endothelial cells appeared necrotic, highly attenuated, and with dense cytoplasm. We showed that epsilon-toxin, in a time- and dose-dependent manner, rapidly and irreversibly compromised the barrier function of venular microvessel endothelium. The results conformed to the hypothesis that epsilon-toxin interacts with vascular endothelial cells and increases the vessel wall permeability by direct damage of the endothelium. PMID:16041001

  6. 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

  7. 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

  8. Clostridium perfringens epsilon toxin H149A mutant as a platform for receptor binding studies

    PubMed Central

    Bokori-Brown, Monika; Kokkinidou, Maria C; Savva, Christos G; Fernandes da Costa, Sérgio; Naylor, Claire E; Cole, Ambrose R; Moss, David S; Basak, Ajit K; Titball, Richard W

    2013-01-01

    Clostridium perfringens epsilon toxin (Etx) is a pore-forming toxin responsible for a severe and rapidly fatal enterotoxemia of ruminants. The toxin is classified as a category B bioterrorism agent by the U.S. Government Centres for Disease Control and Prevention (CDC), making work with recombinant toxin difficult. To reduce the hazard posed by work with recombinant Etx, we have used a variant of Etx that contains a H149A mutation (Etx-H149A), previously reported to have reduced, but not abolished, toxicity. The three-dimensional structure of H149A prototoxin shows that the H149A mutation in domain III does not affect organisation of the putative receptor binding loops in domain I of the toxin. Surface exposed tyrosine residues in domain I of Etx-H149A (Y16, Y20, Y29, Y30, Y36 and Y196) were mutated to alanine and mutants Y30A and Y196A showed significantly reduced binding to MDCK.2 cells relative to Etx-H149A that correlated with their reduced cytotoxic activity. Thus, our study confirms the role of surface exposed tyrosine residues in domain I of Etx in binding to MDCK cells and the suitability of Etx-H149A for further receptor binding studies. In contrast, binding of all of the tyrosine mutants to ACHN cells was similar to that of Etx-H149A, suggesting that Etx can recognise different cell surface receptors. In support of this, the crystal structure of Etx-H149A identified a glycan (?-octyl-glucoside) binding site in domain III of Etx-H149A, which may be a second receptor binding site. These findings have important implications for developing strategies designed to neutralise toxin activity. PMID:23504825

  9. A tripartite cocktail of chimeric monoclonal antibodies passively protects mice against ricin, staphylococcal enterotoxin B and Clostridium perfringens epsilon toxin.

    PubMed

    Sully, Erin K; Whaley, Kevin; Bohorova, Natasha; Bohorov, Ognian; Goodman, Charles; Kim, Do; Pauly, Michael; Velasco, Jesus; Holtsberg, Frederick W; Stavale, Eric; Aman, M Javad; Tangudu, Chandra; Uzal, Francisco A; Mantis, Nicholas J; Zeitlin, Larry

    2014-12-15

    Due to the fast-acting nature of ricin, staphylococcal enterotoxin B (SEB), and Clostridium perfringens epsilon toxin (ETX), it is necessary that therapeutic interventions following a bioterrorism incident by one of these toxins occur as soon as possible after intoxication. Moreover, because the clinical manifestations of intoxication by these agents are likely to be indistinguishable from each other, especially following aerosol exposure, we have developed a cocktail of chimeric monoclonal antibodies that is capable of neutralizing all three toxins. The efficacy of this cocktail was demonstrated in mouse models of lethal dose toxin challenge. PMID:25260254

  10. Proteolytic processing and activation of Clostridium perfringens epsilon toxin by caprine small intestinal contents.

    PubMed

    Freedman, John C; Li, Jihong; Uzal, Francisco A; McClane, Bruce A

    2014-01-01

    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. Importance: Processing and activation by intestinal proteases is a prerequisite for ETX-induced toxicity. Previous studies had characterized the activation of ETX using only arbitrarily chosen amounts of purified trypsin and/or chymotrypsin. Therefore, the current study examined ETX activation ex vivo by natural host intestinal contents. These analyses demonstrated that (i) ETX processing in host intestinal contents occurs in an ordered, stepwise fashion, (ii) processing of prototoxin by host intestinal contents results in higher-molecular-mass material and 3 distinct ~27-kDa ETX species, and (iii) serine proteases, such as trypsin, chymotrypsin, and other proteases, including carboxypeptidases, play a role in the activation of ETX by intestinal contents. These studies provide new insights into the activation and processing of ETX and demonstrate that this process is more complicated than previously appreciated. PMID:25336460

  11. Epsilon toxin is essential for the virulence of Clostridium perfringens type D infection in sheep, goats, and mice.

    PubMed

    Garcia, J P; Adams, V; Beingesser, J; Hughes, M L; Poon, R; Lyras, D; Hill, A; McClane, B A; Rood, J I; Uzal, F A

    2013-07-01

    Clostridium perfringens type D causes disease in sheep, goats, and other ruminants. Type D isolates produce, at minimum, alpha and epsilon (ETX) toxins, but some express up to five different toxins, raising questions about which toxins are necessary for the virulence of these bacteria. We evaluated the contribution of ETX to C. perfringens type D pathogenicity in an intraduodenal challenge model in sheep, goats, and mice using a virulent C. perfringens type D wild-type strain (WT), an isogenic ETX null mutant (etx mutant), and a strain where the etx mutation has been reversed (etx complemented). All sheep and goats, and most mice, challenged with the WT isolate developed acute clinical disease followed by death in most cases. Sheep developed various gross and/or histological changes that included edema of brain, lungs, and heart as well as hydropericardium. Goats developed various effects, including necrotizing colitis, pulmonary edema, and hydropericardium. No significant gross or histological abnormalities were observed in any mice infected with the WT strain. All sheep, goats, and mice challenged with the isogenic etx mutant remained clinically healthy for ?24 h, and no gross or histological abnormalities were observed in those animals. Complementation of etx knockout restored virulence; most goats, sheep, and mice receiving this complemented mutant developed clinical and pathological changes similar to those observed in WT-infected animals. These results indicate that ETX is necessary for type D isolates to induce disease, supporting a key role for this toxin in type D disease pathogenesis. PMID:23630957

  12. 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

  13. Clostridium perfringens epsilon toxin mutant Y30A-Y196A as a recombinant vaccine candidate against enterotoxemia.

    PubMed

    Bokori-Brown, Monika; Hall, Charlotte A; Vance, Charlotte; Fernandes da Costa, Sérgio P; Savva, Christos G; Naylor, Claire E; Cole, Ambrose R; Basak, Ajit K; Moss, David S; Titball, Richard W

    2014-05-13

    Epsilon toxin (Etx) is a ?-pore-forming toxin produced by Clostridium perfringens toxinotypes B and D and plays a key role in the pathogenesis of enterotoxemia, a severe, often fatal disease of ruminants that causes significant economic losses to the farming industry worldwide. This study aimed to determine the potential of a site-directed mutant of Etx (Y30A-Y196A) to be exploited as a recombinant vaccine against enterotoxemia. Replacement of Y30 and Y196 with alanine generated a stable variant of Etx with significantly reduced cell binding and cytotoxic activities in MDCK.2 cells relative to wild type toxin (>430-fold increase in CT50) and Y30A-Y196A was inactive in mice after intraperitoneal administration of trypsin activated toxin at 1000× the expected LD50 dose of trypsin activated wild type toxin. Moreover, polyclonal antibody raised in rabbits against Y30A-Y196A provided protection against wild type toxin in an in vitro neutralisation assay. These data suggest that Y30A-Y196A mutant could form the basis of an improved recombinant vaccine against enterotoxemia. PMID:24709588

  14. Interaction of Clostridium perfringens epsilon-toxin with biological and model membranes: A putative protein receptor in cells.

    PubMed

    Manni, Marco M; Sot, Jesús; Goñi, Félix M

    2015-03-01

    Epsilon-toxin (ETX) is a powerful toxin produced by some strains of Clostridium perfringens (classified as types B and D) that is responsible for enterotoxemia in animals. ETX forms pores through the plasma membrane of eukaryotic cells, consisting of a ?-barrel of 14 amphipathic ?-strands. ETX shows a high specificity for certain cell lines, of which Madin-Darby canine kidney (MDCK) is the first sensitive cell line identified and the most studied one. The aim of this study was to establish the role of lipids in the toxicity caused by ETX and the correlation of its activity in model and biological membranes. In MDCK cells, using cell counting and confocal microscopy, we have observed that the toxin causes cell death mediated by toxin binding to plasma membrane. Moreover, ETX binds and permeabilizes the membranes of giant plasma membrane vesicles (GPMV). However, little effect is observed on protein-free vesicles. The data suggest the essential role of a protein receptor for the toxin in cell membranes. PMID:25485476

  15. Loss of endothelial barrier antigen immunoreactivity as a marker of Clostridium perfringens type D epsilon toxin-induced microvascular damage in rat brain.

    PubMed

    Finnie, J W; Manavis, J; Chidlow, G

    2014-01-01

    The epsilon toxin elaborated by Clostridium perfringens type D in the intestine of domestic livestock is principally responsible for the neurological disease produced after its absorption in excessive quantities into the systemic circulation. The fundamental basis of the cerebral damage induced by epsilon toxin appears to be microvascular injury with ensuing severe, diffuse vasogenic oedema. Endothelial barrier antigen (EBA), which is normally expressed by virtually all capillaries and venules in the rat brain, was used in this study as a marker of blood-brain barrier (BBB) integrity. After exposure to high levels of circulating epsilon toxin, there was substantial loss of EBA in many brain microvessels, attended by widespread plasma albumin extravasation. These results support microvascular injury and subsequent BBB breakdown as a key factor in the pathogenesis of epsilon toxin-induced neurological disease. PMID:24973988

  16. Comparison of four techniques for the detection of Clostridium perfringens type D epsilon toxin in intestinal contents and other body fluids of sheep and goats.

    PubMed

    Uzal, F A; Kelly, W R; Thomas, R; Hornitzky, M; Galea, F

    2003-03-01

    Polyclonal capture enzyme-linked immunosorbent assay (PC-ELISA), monoclonal capture ELISA (MC-ELISA), mouse neutralization test (MNT), and counterimmunoelectrophoresis (CIEP), were compared for their ability to detect epsilon toxin in intestinal contents and body fluids of sheep and goats. When used to evaluate intestinal contents of sheep artificially spiked with epsilon prototoxin, PC-ELISA detected 0.075 mouse lethal dose (MLD)50/ml, whereas the MNT, MC-ELISA, and CIEP detected 6, 25, and 50 MLD50/ml, respectively. Amounts of epsilon toxin detected by PC-ELISA, MC-ELISA, MNT, and CIEP in sheep pericardial fluid artificially spiked with epsilon prototoxin were 0.075, 0.75, 6, and 200 MLD50/ml, respectively. For assaying epsilon toxin in aqueous humor, PC-ELISA and MC-ELISA detected 0.075 MLD50/ml, whereas CIEP detected 200 MLD50/ml (MNT was not evaluated). When 51 samples of intestinal contents of sheep and goats (32 positive and 19 negative to MNT) were analyzed by the other 3 techniques, the relative sensitivity of PC-ELISA, MC-ELISA, and CIEP was 93.75, 84.37, and 37.50%, respectively. The specificity of PC-ELISA, MC-ELISA, and CIEP was 31.57, 57.89, and 84.21%, respectively. The absolute sensitivity of PC-ELISA, MC-ELISA, CIEP, and MNT was 90.90, 69.69, 15.15, and 54.54%. The absolute specificity of the 4 techniques was 100%. These results show that there is a marked inconsistency among techniques routinely used to detect Clostridium perfringens epsilon toxin. Until more consistent results are achieved, the diagnosis of enterotoxemia should not only be based solely on epsilon toxin detection, but also on clinical and pathological data. PMID:12661718

  17. 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

  18. Prevention and treatment of Clostridium perfringens epsilon toxin intoxication in mice with a neutralizing monoclonal antibody (c4D7) produced in Nicotiana benthamiana.

    PubMed

    Garcia, J P; Beingesser, J; Bohorov, O; Bohorova, N; Goodman, C; Kim, D; Pauly, M; Velasco, J; Whaley, K; Zeitlin, L; Roy, C J; Uzal, F A

    2014-09-01

    Epsilon toxin (ETX), produced by Clostridium perfringens types B and D, is among the most lethal toxins known. ETX is a potential bioterrorism threat that was listed as a Category B agent by the U.S. Centers for Disease Control until 2012 and it still remains a toxin of interest for several government agencies. We produced a monoclonal antibody (MAb) against ETX (ETX MAb c4D7) in Nicotiana benthamiana and characterized its preventive and therapeutic efficacy in mice. The ETX preparation used was highly lethal for mice (LD50 = 1.6 ?g/kg) and resulted in a mean time from inoculation to death of 18 and 180 min when administered intravenously or intraperitoneally, respectively. High lethal challenge resulted in dramatic increases of a variety of pro-inflammatory cytokines in serum, while lower, but still lethal doses, did not elicit such responses. ETX MAb c4D7 was highly effective prophylactically (ED50 = 0.3 mg/kg; ED100 = 0.8 mg/kg) and also provided protection when delivered 15-30 min post-ETX intoxication. These data suggest that ETX MAb c4D7 may have use as a pre- and post-exposure treatment for ETX intoxication. PMID:24950050

  19. 9 CFR 113.454 - Clostridium Perfringens Type C Antitoxin.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...ORGANISMS AND VECTORS STANDARD REQUIREMENTS Antibody Products § 113.454 Clostridium Perfringens...Clostridium Perfringens Type C Antitoxin is a specific antibody product containing antibodies directed against the toxin of Clostridium...

  20. 9 CFR 113.455 - Clostridium Perfringens Type D Antitoxin.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...ORGANISMS AND VECTORS STANDARD REQUIREMENTS Antibody Products § 113.455 Clostridium Perfringens...Clostridium Perfringens Type D Antitoxin is a specific antibody product containing antibodies directed against the toxin of Clostridium...

  1. Lytic Clostridium perfringens Bacteriophage 39-O Genomic

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Screening for bacteriophages lytic for Clostridium perfringens was completed utilizing filtered samples obtained from poultry (intestinal material), soil, sewage and poultry processing drainage water. Following limit dilution cloning and three rounds of plaque purification lytic phage preparations ...

  2. 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

  3. Comparative Analysis of Clostridium perfringens Bacteriophage

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  4. 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 ...

  5. Enterotoxigenic Clostridium perfringens: Detection and Identification

    PubMed Central

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

    2012-01-01

    Recent advances in understanding the genetics of enterotoxigenic Clostridium perfringens, including whole genome sequencing of a chromosomal cpe strain and sequencing of several cpe-carrying large plasmids, have led to the development of molecular approaches to more precisely investigate isolates involved in human gastrointestinal diseases and isolates present in the environment. Sequence-based PCR genotyping of the cpe locus (cpe genotyping PCR assays) has provided new information about cpe-positive type A C. perfringens including: 1) Foodborne C. perfringens outbreaks can be caused not only by chromosomal cpe type A strains with extremely heat-resistant spores, but also less commonly by less heat-resistant spore-forming plasmid cpe type A strains; 2) Both chromosomal cpe and plasmid cpe C. perfringens type A strains can be found in retail foods, healthy human feces and the environment, such as in sewage; 3) Most environmental cpe-positive C. perfringens type A strains carry their cpe gene on plasmids. Moreover, recent studies indicated that the cpe loci of type C, D, and E strains differ from the cpe loci of type A strains and from the cpe loci of each other, indicating that the cpe loci of C. perfringens have remarkable diversity. Multi-locus sequence typing (MLST) indicated that the chromosomal cpe strains responsible for most food poisoning cases have distinct genetic characteristics that provide unique biological properties, such as the formation of highly heat-resistant spores. These and future advances should help elucidate the epidemiology of enterotoxigenic C. perfringens and also contribute to the prevention of C. perfringens food poisoning outbreaks and other CPE-associated human diseases. PMID:22504431

  6. Clostridium difficile and Clostridium perfringens from wild carnivore species in Brazil.

    PubMed

    Silva, Rodrigo Otávio Silveira; D'Elia, Mirella Lauria; Tostes Teixeira, Erika Procópio; Pereira, Pedro Lúcio Lithg; de Magalhães Soares, Danielle Ferreira; Cavalcanti, Álvaro Roberto; Kocuvan, Aleksander; Rupnik, Maja; Santos, André Luiz Quagliatto; Junior, Carlos Augusto Oliveira; Lobato, Francisco Carlos Faria

    2014-08-01

    Despite some case reports, the importance of Clostridium perfringens and Clostridium difficile for wild carnivores remains unclear. Thus, the objective of this study was to identify C. perfringens and C. difficile strains in stool samples from wild carnivore species in Brazil. A total of 34 stool samples were collected and subjected to C. perfringens and C. difficile isolation. Suggestive colonies of C. perfringens were then analyzed for genes encoding the major C. perfringens toxins (alpha, beta, epsilon and iota) and the beta-2 toxin (cpb2), enterotoxin (cpe) and NetB (netb) genes. C. difficile strains were analyzed by multiplex-PCR for toxins A (tcdA) and B (tcdB) and a binary toxin gene (cdtB) and also submitted to a PCR ribotyping. Unthawed aliquots of samples positive for C. difficile isolation were subjected to the detection of A/B toxins by a cytotoxicity assay (CTA). C. perfringens was isolated from 26 samples (76.5%), all of which were genotyped as type A. The netb gene was not detected, whereas the cpb2 and cpe genes were found in nine and three C. perfringens strains, respectively. C. difficile was isolated from two (5.9%) samples. A non-toxigenic strain was recovered from a non-diarrheic maned wolf (Chrysocyon brachyurus). Conversely, a toxigenic strain was found in the sample of a diarrheic ocelot (Leopardus pardallis); an unthawed stool sample was also positive for A/B toxins by CTA, indicating a diagnosis of C. difficile-associated diarrhea in this animal. The present work suggests that wild carnivore species could carry C. difficile strains and that they could be susceptible to C. difficile infection. PMID:24979683

  7. Clostridium Perfringens Toxins Involved in Mammalian Veterinary Diseases

    PubMed Central

    Uzal, F. A.; Vidal, J. E.; McClane, B. A.; Gurjar, A. A.

    2013-01-01

    Clostridium perfringens is a gram-positive anaerobic rod that is classified into 5 toxinotypes (A, B, C, D, and E) according to the production of 4 major toxins, namely alpha (CPA), beta (CPB), epsilon (ETX) and iota (ITX). However, this microorganism can produce up to 16 toxins in various combinations, including lethal toxins such as perfringolysin O (PFO), enterotoxin (CPE), and beta2 toxin (CPB2). Most diseases caused by this microorganism are mediated by one or more of these toxins. The role of CPA in intestinal disease of mammals is controversial and poorly documented, but there is no doubt that this toxin is essential in the production of gas gangrene of humans and several animal species. CPB produced by C. perfringens types B and C is responsible for necrotizing enteritis and enterotoxemia mainly in neonatal individuals of several animal species. ETX produced by C. perfringens type D is responsible for clinical signs and lesions of enterotoxemia, a predominantly neurological disease of sheep and goats. The role of ITX in disease of animals is poorly understood, although it is usually assumed that the pathogenesis of intestinal diseases produced by C. perfringens type E is mediated by this toxin. CPB2, a necrotizing and lethal toxin that can be produced by all types of C. perfringens, has been blamed for disease in many animal species, but little information is currently available to sustain or rule out this claim. CPE is an important virulence factor for C. perfringens type A gastrointestinal disease in humans and dogs; however, the data implicating CPE in other animal diseases remains ambiguous. PFO does not seem to play a direct role as the main virulence factor for animal diseases, but it may have a synergistic role with CPA-mediated gangrene and ETX-mediated enterotoxemia. The recent improvement of animal models for C. perfringens infection and the use of toxin gene knock-out mutants have demonstrated the specific pathogenic role of several toxins of C. perfringens in animal disease. These research tools are helping us to establish the role of each C. perfringens toxin in animal disease, to investigate the in vivo mechanism of action of these toxins, and to develop more effective vaccines against diseases produced by these microorganisms. PMID:24511335

  8. 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.

  9. Towards an understanding of the role of Clostridium perfringens toxins in human and animal disease

    PubMed Central

    Uzal, Francisco A; Freedman, John C; Shrestha, Archana; Theoret, James R; Garcia, Jorge; Awad, Milena M; Adams, Vicki; Moore, Robert J; Rood, Julian I; McClane, Bruce A

    2014-01-01

    Clostridium perfringens uses its arsenal of >16 toxins to cause histotoxic and intestinal infections in humans and animals. It has been unclear why this bacterium produces so many different toxins, especially since many target the plasma membrane of host cells. However, it is now established that C. perfringens uses chromosomally encoded alpha toxin (a phospholipase C) and perfringolysin O (a pore-forming toxin) during histotoxic infections. In contrast, this bacterium causes intestinal disease by employing toxins encoded by mobile genetic elements, including C. perfringens enterotoxin, necrotic enteritis toxin B-like, epsilon toxin and beta toxin. Like perfringolysin O, the toxins with established roles in intestinal disease form membrane pores. However, the intestinal disease-associated toxins vary in their target specificity, when they are produced (sporulation vs vegetative growth), and in their sensitivity to intestinal proteases. Producing many toxins with diverse characteristics likely imparts virulence flexibility to C. perfringens so it can cause an array of diseases. PMID:24762309

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

    PubMed

    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

  11. Distribution of Clostridium perfringens isolates from piglets in South Korea.

    PubMed

    Lee, Ki-Eun; Lim, Seong-In; Shin, Seong-Ho; Kwon, Yong-Kuk; Kim, Ha-Young; Song, Jae-Young; An, Dong-Jun

    2014-05-01

    Clostridium perfringens causes various digestive system disease symptoms in pigs. In the present study, 11 C. perfringens isolates were obtained from diarrheic piglets and 18 from healthy piglets. All of the C. perfringens isolates were shown to be type A using a multiplex PCR assay. The ?2 toxin gene was detected in 27/29 C. perfringens isolates, i.e., 81% (9/11) of diarrheic piglets and 100% (18/18) of healthy piglets, and all of the genes had the same sequence. In conclusion, the ?2 toxin gene of C. perfringens was distributed widely in Korean piglets regardless of the incidence of diarrhea, and there was no clear relationship with enteric disease. A pulsed-field gel electrophoresis analysis of DNA digested using SmaI demonstrated the non-clonal spread of C. perfringens isolates from piglets. PMID:24430655

  12. 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 ...

  13. Translational selection is operative for synonymous codon usage in Clostridium perfringens and Clostridium acetobutylicum

    Microsoft Academic Search

    Hector Musto; Hector Romero; Alejandro Zavala

    2003-01-01

    Here, the codon usage patterns of two Clostridium species (Clostridium perfringens and Clostridium acetobutylicum) are reported. These prokaryotes are characterized by a strong mutational bias towards A+T, a striking excess of coding sequences and purine-rich leading strands of replication, strong GC-skews and a high frequency of genomic rearrangements. As expected, it was found that the mutational bias dominates codon usage

  14. Tips to Prevent Illness from Clostridium Perfringens

    MedlinePLUS

    ... is one of the most common causes of food poisoning in the United States. Learn more on ways ... 1 million cases of foodborne illness (sometimes called "food poisoning") each year. C. perfringens is found in many ...

  15. Clostridium perfringens type A-E toxin plasmids.

    PubMed

    Freedman, John C; Theoret, James R; Wisniewski, Jessica A; Uzal, Francisco A; Rood, Julian I; McClane, Bruce A

    2014-10-01

    Clostridium perfringens relies upon plasmid-encoded toxin genes to cause intestinal infections. These toxin genes are associated with insertion sequences that may facilitate their mobilization and transfer, giving rise to new toxin plasmids with common backbones. Most toxin plasmids carry a transfer of clostridial plasmids locus mediating conjugation, which likely explains the presence of similar toxin plasmids in otherwise unrelated C. perfringens strains. The association of many toxin genes with insertion sequences and conjugative plasmids provides virulence flexibility when causing intestinal infections. However, incompatibility issues apparently limit the number of toxin plasmids maintained by a single cell. PMID:25283728

  16. 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

    2014-10-16

    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

  17. Cross-complementation of Clostridium perfringens PLC and Clostridium septicum ?-toxin mutants reveals PLC is sufficient to mediate gas gangrene

    Microsoft Academic Search

    Catherine L. Kennedy; Dena Lyras; Jackie K. Cheung; Thomas J. Hiscox; John J. Emmins; Julian I. Rood

    2009-01-01

    Clostridium perfringens and Clostridium septicum are the most common causes of clostridial myonecrosis or gas gangrene. Although they mediate a similar disease pathology, they elaborate functionally very different ?-toxins. We used a reciprocal complementation approach to assess the contribution of the primary toxin of each species to disease and found that C. perfringens ?-toxin (PLC) was able to mediate the

  18. Clostridium perfringens Sepsis and Fetal Demise after Genetic Amniocentesis

    PubMed Central

    Hendrix, Nancy W.; Mackeen, A. Dhanya; Weiner, Stuart

    2011-01-01

    Clostridium perfringens is a rare cause of intrauterine infection. There have been five case reports concerning infection associated with invasive procedures. We report a woman who underwent a genetic amniocentesis due to her history of chronic granulomatous disease. She presented to the hospital ?38 hours after the amniocentesis complaining of fever and chills. Due to acute decompensation, she underwent an emergent dilatation and evacuation. During her stay, blood cultures came back positive for C. perfringens. Gradual improvement with intensive monitoring led to hospital discharge 4 days after the procedure. Uterine infection due to C. perfringens leading to maternal sepsis is associated with a high morbidity and mortality rate. Our patient was able to survive without a hysterectomy due to the rapid administration of antibiotics and surgical intervention while being evaluated. PMID:23705080

  19. Hazard analysis of Clostridium perfringens in the Skylab Food System

    NASA Technical Reports Server (NTRS)

    Bourland, C. T.; Huber, C. S.; Kiser, P. R.; Heidelbaugh, N. D.; Rowley, D. B.

    1974-01-01

    The Skylab Food System presented unique microbiological problems because food was warmed in null-gravity and because the heat source was limited to 69.4 C (to prevent boiling in null-gravity). For these reasons, the foods were manufactured using critical control point techniques of quality control coupled with appropriate hazard analyses. One of these hazard analyses evaluated the threat from Clostridium perfringens. Samples of food were inoculated with C. perfringens and incubated for 2 h at temperatures ranging from 25 to 55 C. Generation times were determined for the foods at various temperatures. Results of these tests were evaluated taking into consideration: food-borne disease epidemiology, the Skylab food manufacturing procedures, and the performance requirements of the Skylab Food System. Based on this hazard analysis, a limit for C. perfringens of 100/g was established for Skylab foods.

  20. Molecular Subtyping of Poultry-Associated Type A Clostridium perfringens Isolates by Repetitive-Element PCR

    PubMed Central

    Siragusa, G. R.; Danyluk, M. D.; Hiett, K. L.; Wise, M. G.; Craven, S. E.

    2006-01-01

    Clostridium perfringens strains (type A) isolated from an integrated poultry operation were subtyped using repetitive-element PCR with Dt primers. Isolates were obtained from fecal, egg shell, fluff, and carcass rinse samples as part of a previously reported temporally linked epidemiological survey. A total of 48 isolates of C. perfringens were obtained from different stages of the broiler chicken production chain from two separate breeder farms that supplied a single hatchery that in turn provided chicks to a single grow-out farm whose flocks were processed at a single plant. All 48 isolates were typeable (100% typeability) by repetitive-element PCR with Dt primers. This subtyping method was highly reproducible and discriminatory. By repetitive-element PCR with Dt primers, isolates were classified into four major branches with 12 subgroups or clades. The Simpson's index of discrimination was calculated to be 0.96 for groupings of >95% correlation. Toxin gene profiles of the isolates indicated that all of the isolates were C. perfringens alpha-toxin gene positive and 46 of 48 isolates were beta2-toxin gene positive. All strains were negative for beta- and epsilon-toxin genes. Repetitive sequence-based PCR was found to be a technically practical and reproducible means of subtyping C. perfringens libraries from specific epidemiological or production environment settings. PMID:16517895

  1. 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

  2. Role of Clostridium perfringens and Clostridium septicum in causing turkey cellulitis.

    PubMed

    Thachil, Anil J; McComb, Brian; Andersen, Michelle M; Shaw, Daniel P; Halvorson, David A; Nagaraja, Kakambi V

    2010-06-01

    The role of Clostridium perfringens and Clostridium septicum in the development of cellulitis and mortality in turkey poults was examined. Studies were done in turkeys of two age groups: 3-wk-old and 7-wk-old turkey poults. The effect of varying doses of C. perfringens and C. septicum in reproducing cellulitis lesions and mortality in turkeys was investigated. Both in vitro and in vivo assays were conducted to study their toxic and biologic activities. Clostridium septicum spore culture was found to be more potent than that of C. perfringens in both in vitro assays, such as the hemolysis test, and in vivo assays in mice and turkeys. Both C. perfringens and C. septicum spore cultures were found to be capable of inducing cellulitis lesions and mortality in turkey poults when inoculated by subcutaneous route. Histopathology examination of affected tissues revealed a "moth-eaten appearance, with abundant growth of C. perfringens and C. septicum in the sarcomeres of muscle tissues and in the subcutaneous tissues. However, C. septicum was found to be more potent than C. perfringens in causing cellulitis lesions and mortality in turkeys. Three-week-old poults were found to be less susceptible than 7-wk-old poults in the development of cellulitis lesions and mortality after inoculation with either spore cultures of C. perfringens or C. septicum. The results of the current study suggest that although C. septicum is more potent in causing cellulitis lesions and mortality, infection with either C. septicum or C. perfringens can cause cellulitis lesions and mortality in turkeys. PMID:20608521

  3. 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

  4. Detection and molecular typing of Clostridium perfringens isolates from beef, chicken and turkey meats.

    PubMed

    Aras, Zeki; Hadimli, Hasan Hüseyin

    2015-04-01

    Here we describe a study investigating the presence of Clostridium perfringens strains in meat samples and the toxin genes in the isolates by PCR. This study, for the first time, demonstrated the presence of C. perfringens type E in turkey meats, while C. perfringens type C strains isolated from chicken meats. PMID:25460196

  5. Screening Various Plant and Fungal Extracts Against Clostridium perfringens for Antimicrobial Activity

    Microsoft Academic Search

    Thomas R. Larson

    2007-01-01

    Clostridium is an anaerobic, endospore forming Gram-positive bacillus genus containing many important pathogenic species, including Clostridium difficile and Clostridium perfringens. Both of these pathogenic species have been implicated as the causative agents of serious gastrointestinal tract infections. Recently, there has been an emergence of antimicrobial drug resistant Clostridium species. Because of these concerns, a new drug is warranted. There is

  6. Development of an Elisa Assay for Clostridium Perfringens Phospholipase C (Alpha Toxin)

    Microsoft Academic Search

    R. J. Holdsworth; D. Parratt

    1994-01-01

    A new method for the assay of Clostridium perfringens alpha toxin (phospholipase C) is described using a sandwich ELISA. This assay has been shown to be quantitative, to have a high specificity for the toxin and is capable of detecting purified Clostridium perfringens phospholipase C at concentrations of as little as 0.005 units\\/ml in cooked meat culture medium.

  7. 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...

  8. Occurrence of Clostridium perfringens from different cultivated soils.

    PubMed

    Voidarou, C; Bezirtzoglou, E; Alexopoulos, A; Plessas, S; Stefanis, C; Papadopoulos, I; Vavias, S; Stavropoulou, E; Fotou, K; Tzora, A; Skoufos, I

    2011-12-01

    The occurrence of Clostridium perfringens was estimated in 750 samples originated from a variety of soils bearing various bulb crops: Brawnica oderacea (vegetable), Olea europaea, Daucus carota (carote), Solanum tuberosum (potato), Phaseolus vulgaris (green haricot), Beta vulgaris var. rapaceum (beetroot), Cucurbita pepo (squash), Allium cepa (onion), Cucumis sativus (cucumber) and Capsicum annum (pepper). All isolated strains were tested for their antimicrobial activities to amoxicillin, penicillin G, kanamycin, tetracycline, streptomycin, erythromycin, chloramphenicol and metronidazole. When considering the type of the bulb production, it was observed increased number of C. perfringens spore densities in the most undersurface bulb soils. Moreover, C. perfringens spore are likely to occur in particularly large numbers in soil contaminated by fecal matter. Additionally, there is a close relationship between the spore amount and nature of organic content. Presence of C. perfringens was associated with acidic soil. Most of our strains showed resistance to the studied antibiotics applied usually for human and veterinary care. A systematic monitoring of the cultivated soil ecosystems must include bacteriological parameters together with chemical indices of organic pollution in order to obtain information adequate for assessing their overall quality. PMID:21621626

  9. Recent Insights into Clostridium perfringens Beta-Toxin

    PubMed Central

    Nagahama, Masahiro; Ochi, Sadayuki; Oda, Masataka; Miyamoto, Kazuaki; Takehara, Masaya; Kobayashi, Keiko

    2015-01-01

    Clostridium perfringens beta-toxin is a key mediator of necrotizing enterocolitis and enterotoxemia. It is a pore-forming toxin (PFT) that exerts cytotoxic effect. Experimental investigation using piglet and rabbit intestinal loop models and a mouse infection model apparently showed that beta-toxin is the important pathogenic factor of the organisms. The toxin caused the swelling and disruption of HL-60 cells and formed a functional pore in the lipid raft microdomains of sensitive cells. These findings represent significant progress in the characterization of the toxin with knowledge on its biological features, mechanism of action and structure-function having been accumulated. Our aims here are to review the current progresses in our comprehension of the virulence of C. perfringens type C and the character, biological feature and structure-function of beta-toxin. PMID:25654787

  10. Complication of Invasive Molar Pregnancy with Clostridium perfringens Sepsis.

    PubMed

    Singh, Sanmeet; Angra, Kunal; Davis, Bonnie; Shokrani, Babak

    2014-01-01

    Clostridium perfringens (CP) is an anaerobic, Gram-positive bacillus associated with malignant diseases and near-term pregnancies. The necrotic tissue that results from these disease processes fuels the proliferation of CP, leading to gas gangrene and subsequently sepsis. Herein, we report a case of a 41-year-old female patient with a history of invasive molar pregnancy that was further complicated with a CP infection. Although past research has shown a link between Clostridium infection and choriocarcinoma (Chern-Horng and Hsieh, 1999), no previous cases of CP infection have been associated with invasive molar pregnancy. We also report complete resolution of the CP sepsis and its associated symptoms following the hysterectomy. PMID:24716030

  11. Complication of Invasive Molar Pregnancy with Clostridium perfringens Sepsis

    PubMed Central

    Singh, Sanmeet; Angra, Kunal; Davis, Bonnie; Shokrani, Babak

    2014-01-01

    Clostridium perfringens (CP) is an anaerobic, Gram-positive bacillus associated with malignant diseases and near-term pregnancies. The necrotic tissue that results from these disease processes fuels the proliferation of CP, leading to gas gangrene and subsequently sepsis. Herein, we report a case of a 41-year-old female patient with a history of invasive molar pregnancy that was further complicated with a CP infection. Although past research has shown a link between Clostridium infection and choriocarcinoma (Chern-Horng and Hsieh, 1999), no previous cases of CP infection have been associated with invasive molar pregnancy. We also report complete resolution of the CP sepsis and its associated symptoms following the hysterectomy. PMID:24716030

  12. Regulation of toxin gene expression in Clostridium perfringens.

    PubMed

    Ohtani, Kaori; Shimizu, Tohru

    2014-10-01

    The Gram-positive, anaerobic, spore-forming, rod-shaped Clostridium perfringens is widely distributed in nature, especially in soil and the gastrointestinal tract of humans and animals. C. perfringens causes clostridial myonecrosis (or gas gangrene), enteritis and enterotoxemia in humans and livestock by producing numerous extracellular toxins and enzymes. The toxin gene expression is regulated by a two-component regulatory system and regulatory RNA VirR/VirS-VR-RNA cascade. The VirR/VirS system was originally found in a type A strain, but a recent report showed that it is also important for the toxin gene regulation in other types of strains. Two types of cell-cell signaling, i.e., agr-system and AI-2 signaling, are also important for the regulation of toxin genes. Several regulatory systems independent from the VirR/VirS system, including virX, the orphan histidine kinase ReeS and orphan response regulator RevR, are also involved in the regulation of toxin genes. In addition, the expression of toxin genes is upregulated after contact with Caco-2 cells. C. perfringens has a complex regulatory network for toxin gene expression and thus the coordination of toxin gene expression is important for the process of infection. PMID:25303832

  13. Clostridium perfringens enterotoxicosis in two Amur leopards (Panthera pardus orientalis).

    PubMed

    Neiffer, D L

    2001-03-01

    Two 6-yr-old male sibling Amur leopards (Panthera pardus orientalis) housed together at the Pittsburgh Zoo presented for acute onset of diarrhea with no changes in appetite or behavior. Heat-fixed modified Wright-stained and Gram-stained fecal smears revealed a mixed bacterial population with a large number of gram-positive Clostridium perfringens-like spores (>20 per high-power oil immersion field). In addition, C. perfringens enterotoxin was isolated from one leopard at 1:256, confirming the presence of C. perfringens enterotoxicosis. Treatment with oral metronidazole, tylosin tartrate, and psyllium fiber was prescribed, with return of more normal stool by the third day of treatment. Fecal consistency steadily improved and was considered normal by the time all prescribed treatments were complete. Diarrhea has not recurred. Partially thawed meat in the leopards' diet may have precipitated the production of an endogenous clostridial enterotoxicosis by disrupting digestive tract flora with resultant clostridial overgrowth and sporulation. PMID:12790411

  14. 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

  15. Effect of cooling on Clostridium perfringens in pea soup.

    PubMed

    de Jong, A E I; Rombouts, F M; Beumer, R R

    2004-02-01

    Foods associated with Clostridium perfringens outbreaks are usually abused after cooking. Because of their short generation times, C. perfringens spores and cells can grow out to high levels during improper cooling. Therefore, the potential of C. perfringens to multiply in Dutch pea soup during different cooling times was investigated. Tubes of preheated pea soup (50 degrees C) were inoculated with cocktails of cells or heat-activated spores of this pathogen. The tubes were linearly cooled to 15 degrees C in time spans of 3, 5, 7.5, and 10 h and were subsequently stored in a refrigerator at 3 or 7 degrees C for up to 84 h. Cell numbers increased by 1-log cycle during the 3-h cooling period and reached their maximum after 10 h of cooling. Subsequent refrigeration hardly reduced cell numbers. Cooling of 3.75 liters of pea soup in an open pan showed that this amount of pea soup cooled from 50 to 15 degrees C in 5 h, which will allow a more than 10-fold increase in cell numbers. These findings emphasize the need of good hygienic practices and quick cooling of heated foods after preparation. PMID:14968969

  16. On the interaction of Clostridium perfringens enterotoxin with claudins.

    PubMed

    Veshnyakova, Anna; Protze, Jonas; Rossa, Jan; Blasig, Ingolf E; Krause, Gerd; Piontek, Joerg

    2010-06-01

    Clostridium perfringens causes one of the most common foodborne illnesses, which is largely mediated by the Clostridium perfringens enterotoxin (CPE). The toxin consists of two functional domains. The N-terminal region mediates the cytotoxic effect through pore formation in the plasma membrane of the mammalian host cell. The C-terminal region (cCPE) binds to the second extracellular loop of a subset of claudins. Claudin-3 and claudin-4 have been shown to be receptors for CPE with very high affinity. The toxin binds with weak affinity to claudin-1 and -2 but contribution of these weak binding claudins to CPE-mediated disease is questionable. cCPE is not cytotoxic, however, it is a potent modulator of tight junctions. This review describes recent progress in the molecular characterization of the cCPE-claudin interaction using mutagenesis, in vitro binding assays and permeation studies. The results promote the development of recombinant cCPE-proteins and CPE-based peptidomimetics to modulate tight junctions for improved drug delivery or to treat tumors overexpressing claudins. PMID:22069641

  17. On the Interaction of Clostridium perfringens Enterotoxin with Claudins

    PubMed Central

    Veshnyakova, Anna; Protze, Jonas; Rossa, Jan; Blasig, Ingolf E.; Krause, Gerd; Piontek, Joerg

    2010-01-01

    Clostridium perfringens causes one of the most common foodborne illnesses, which is largely mediated by the Clostridium perfringens enterotoxin (CPE). The toxin consists of two functional domains. The N-terminal region mediates the cytotoxic effect through pore formation in the plasma membrane of the mammalian host cell. The C-terminal region (cCPE) binds to the second extracellular loop of a subset of claudins. Claudin-3 and claudin-4 have been shown to be receptors for CPE with very high affinity. The toxin binds with weak affinity to claudin-1 and -2 but contribution of these weak binding claudins to CPE-mediated disease is questionable. cCPE is not cytotoxic, however, it is a potent modulator of tight junctions. This review describes recent progress in the molecular characterization of the cCPE-claudin interaction using mutagenesis, in vitro binding assays and permeation studies. The results promote the development of recombinant cCPE-proteins and CPE-based peptidomimetics to modulate tight junctions for improved drug delivery or to treat tumors overexpressing claudins. PMID:22069641

  18. Prevalence and diversity of toxigenic Clostridium perfringens and Clostridium difficile among swine herds in the midwest.

    PubMed

    Baker, Ashley A; Davis, Ellen; Rehberger, Thomas; Rosener, Daniel

    2010-05-01

    Clostridium perfringens and Clostridium difficile are associated with scours in the neonatal piglet and are an economic concern in swine production. The objective of this study was to characterize the prevalence and diversity of C. perfringens and C. difficile isolates obtained from scouring neonatal piglets in a large integrated production system, as well as in smaller independently owned regional farms. Rectal swabs were collected from 333 pigs at 11 sites in an integrated swine production system and from an additional 180 pigs at 16 regional farms located throughout the Midwest. C. perfringens was isolated from 89.8% of the pigs swabbed at the integrated sites, and C. difficile was isolated from 57.7% of these pigs. Of the pigs from the regional farms sampled, 95.6% were positive for isolation of C. perfringens and 27.2% were positive for C. difficile. Toxigenic isolates were typed using random amplified polymorphic DNA (RAPD) PCR, and were placed in four dendrograms for C. perfringens and C. difficile populations isolated from the integrated sites and regional farms. Diversity indices showed that there was greater diversity in C. difficile populations and in populations isolated from the regional farms. A subset of isolates from the C. difficile dendrograms were further toxinotyped by amplification of the pathogenicity locus and subsequent digestion by HincII, AccI, and EcoRI. Of the 45 isolates typed, 44 were determined to be toxinotype V. The results of this study illustrate the diversity of C. perfringens and C. difficile isolates and the prevalence of these pathogens in swine production sites. PMID:20208029

  19. 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...

  20. Phenotypic and genotypic characterization of tetracycline and minocycline resistance in Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aim of this study was to determine the incidence of tetracycline resistance and the prevalence of tetracycline-resistance genes in strains of Clostridium perfringens isolated from different sources between 1994 and 2005. Susceptibility to tetracycline and minocycline in C. perfringens isolates ...

  1. EFFECT OF OZONE STRESS ON CLOSTRIDIUM PERFRINGENS VIABILITY FOLLOWING THE AQUEOUS TREATMENT OF BEEF SURFACES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The antimicrobial efficacy of ozone on the food-borne pathogen, Clostridium perfringens, was evaluated on London Broil top round cut beef surfaces using an aqueous wash system. Current food processing methods do not assure elimination of spores of C. perfringens, thus there is a high likelihood of ...

  2. A role for the Clostridium perfringens ?2 toxin in bovine enterotoxaemia?

    Microsoft Academic Search

    C Manteca; G Daube; T Jauniaux; A Linden; V Pirson; J Detilleux; A Ginter; P Coppe; A Kaeckenbeeck; J. G Mainil

    2002-01-01

    Non-enterotoxigenic type A Clostridium perfringens are associated with bovine enterotoxaemia, but the ? toxin is not regarded as responsible for the production of typical lesions of necrotic and haemorrhagic enteritis. The purpose of this study was to investigate the putative role of the more recently described ?2 toxin. Seven hundred and fourteen non-enterotoxigenic type A C. perfringens isolated from 133

  3. Clostridium perfringens in Long Island Sound sediments: An urban sedimentary record

    USGS Publications Warehouse

    Buchholtz ten Brink, M. R.; Mecray, E.L.; Galvin, E.L.

    2000-01-01

    Clostridium perfringens is a conservative tracer and an indicator of sewage-derived pollution in the marine environment. The distribution of Clostridium perfringens spores was measured in sediments from Long Island Sound, USA, as part of a regional study designed to: (1) map the distribution of contaminated sediments; (2) determine transport and dispersal paths; (3) identify the locations of sediment and contaminant focusing; and (4) constrain predictive models. In 1996, sediment cores were collected at 58 stations, and surface sediments were collected at 219 locations throughout the Sound. Elevated concentrations of Clostridium perfringens in the sediments indicate that sewage pollution is present throughout Long Island Sound and has persisted for more than a century. Concentrations range from undetectable amounts to 15,000 spores/g dry sediment and are above background levels in the upper 30 cm at nearly all core locations. Sediment focusing strongly impacts the accumulation of Clostridium perfringens spores. Inventories in the cores range from 28 to 70,000 spores/cm2, and elevated concentrations can extend to depths of 50 cm. The steep gradients in Clostridium perfringens profiles in muddier cores contrast with concentrations that are generally constant with depth in sandier cores. Clostridium perfringens concentrations rarely decrease in the uppermost sediment, unlike those reported for metal contaminants. Concentrations in surface sediments are highest in the western end of the Sound, very low in the eastern region, and intermediate in the central part. This pattern reflects winnowing and focusing of Clostridium perfringens spores and fine-grained sediment by the hydrodynamic regime; however, the proximity of sewage sources to the westernmost Sound locally enhances the Clostridium perfringens signals.

  4. Molecular typing and epidemiological survey of prevalence of Clostridium perfringens types by multiplex PCR.

    PubMed Central

    Yoo, H S; Lee, S U; Park, K Y; Park, Y H

    1997-01-01

    Clostridium perfringens has been classified into five toxigenic types (A through E) on the basis of its capability to produce major lethal toxins (alpha, beta, epsilon, and iota toxins). Seroneutralization with mice or guinea pigs has been used to type each toxin, but this conventional method has some disadvantages. Therefore, we used a molecular biological technique to type the bacterium in the present study. A multiplex PCR was developed for this purpose. This method has several advantages in comparison with seroneutralization with mice or guinea pigs. By this method, we also investigated the most prevalent type(s) of the organism in Korean calves, piglets, and chickens showing clinical symptoms such as diarrhea, enterotoxemia, and necrotic enteritis. Only type A was isolated from calves and chickens, while type C (2 of 14 isolates), in addition to type A, was isolated from piglets. These results suggested that seroneutralization could be replaced by our new method and that type A of C. perfringens is the most prevalent type in livestock in Korea. PMID:8968913

  5. Cytology of Spore Formation in Clostridium perfringens1

    PubMed Central

    Hoeniger, Judith F. M.; Stuart, Philip F.; Holt, Stanley C.

    1968-01-01

    The sequential morphological events in spore formation by Clostridium perfringens type D were observed in Ellner's medium where 80 to 100% of the cells formed spores. Gross structural changes were studied with the light microscope under phase-contrast, and in fixed cells by the use of both nigrosin and Giemsa preparations. Fine structure was examined with the electron microscope in both thin sections and frozen-etched preparations. During the first 3 hr of incubation, the original rod-shaped cells became ellipsoid to ovoid in shape; by 5 to 6 hr, subterminal spores had developed within these enlarged cells. The fine structural sequence was in most respects identical to that in other Bacillaceae, although some stages were illustrated with particular clarity. A unique feature was the development of a convoluted, membranous exosporium which adhered to the outer surface of the two coats and had an unusual fine structure resembling a rectangular array of subunits. Images PMID:4302300

  6. Massive intravascular hemolysis from Clostridium perfringens septicemia: a review.

    PubMed

    Simon, Tracey G; Bradley, Joanna; Jones, Adisa; Carino, Gerardo

    2014-01-01

    We describe the case of a patient with hemolysis-associated Clostridium perfringens septicemia and review all similar cases published in the literature since 1990, with specific focus on the relationship between treatment strategy and survival. We searched PubMed for all published cases of C. perfringens-associated hemolysis, using the medical subject terms "clostridia," "clostridial sepsis," and/or "hemolysis." All case reports, case series, review articles, and other relevant references published in the English literature since 1990 were included in this study. There were no exclusion criteria. Each case was examined with respect to presenting features of illness, antibiotic regimen, time-to-antibiotic therapy, additional interventions, complications, and patient survival. These variables were entered into a data set and then systematically analyzed with the aid of a statistician, using serial t tests and chi-square analyses. Since 1990, 50 patients of C. perfringens septicemia with hemolysis have been reported. Median age was 61 years (range 31-84), and 58% were male. Mortality was 74%, with a median time to death of 9.7 hours (range 0-96 hours). Of the patients, 35 (70%) were treated medically, while 15 (30%) received antibiotics and surgery. Surgical intervention was associated with significantly improved survival (risk ratio [RR] 0.23, 95% confidence interval [CI] 0.10, 0.53) as was the use of a combination of penicillin and clindamycin (RR of death 0.46, 95% CI 0.25, 0.83). Four patients utilizing hyperbaric oxygen therapy (HBOT) have been reported, and all patients survived. In cases of clostridial sepsis with hemolysis, strong predictors of survival include early initiation of appropriate antibiotics as well as surgical removal of infected foci. The HBOT may also be associated with survival. The disease often progresses rapidly to death, so rapid recognition is critical for the patient survival. PMID:24019300

  7. Comparative Analysis of Prevalence, Risk Factors, and Molecular Epidemiology of Antibiotic-Associated Diarrhea Due to Clostridium difficile, Clostridium perfringens, and Staphylococcus aureus

    Microsoft Academic Search

    N. J. Asha; D. Tompkins; M. H. Wilcox

    2006-01-01

    We prospectively studied the comparative epidemiology and risk factors for Clostridium difficile, Clostridium perfringens, and Staphylococcus aureus antibiotic-associated diarrhea (AAD). Four thousand six hundred fifty- nine inpatient fecal specimens (11 months) were tested for C. difficile cytotoxin, C. perfringens enterotoxin, and S. aureus by Vero cell assay, enzyme-linked immunosorbent assay, and growth on fresh blood agar, respectively. Two distinct age-,

  8. 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

    .......................................................................................... 3 Clostridium perfringens as a foodborne pathogen ........................................ 3 C. perfringens food poisoning.............................................................. 3 Distribution of C. perfringens..., 50, 76). Most of the strains produce ?-toxin (lecithinase, phospholipase C) (50). Only C. perfringens type A and C have been associated with human gastroenteritis (11, 32, 36, 50). C. perfringens food poisoning Gastroenteritis caused by C...

  9. Four Foodborne Disease Outbreaks Caused by a New Type of Enterotoxin-Producing Clostridium perfringens.

    PubMed

    Monma, Chie; Hatakeyama, Kaoru; Obata, Hiromi; Yokoyama, Keiko; Konishi, Noriko; Itoh, Takeshi; Kai, Akemi

    2015-03-01

    The epidemiological and bacteriological investigations on four foodborne outbreaks caused by a new type of enterotoxin-producing Clostridium perfringens are described. C. perfringens isolated from patients of these outbreaks did not produce any known enterotoxin and did not carry the C. perfringens enterotoxin gene. However, the culture filtrates of these isolates induced the accumulation of fluid in rabbit ileal loop tests. The molecular weight of the new enterotoxin may be between 50,000 and 100,000, although the known C. perfringens enterotoxin is ca. 35,000. This new enterotoxin was heat labile, and its biological activities were inactivated by heating for 5 min at 60°C. The new enterotoxin was sensitive to pH values higher than 11.0 and protease treatment but was resistant to trypsin treatment. These results suggest that the new enterotoxin may be a protein. Although C. perfringens enterotoxin induced morphological changes in Vero cells, the changes induced by the new enterotoxin differed from those by the known C. perfringens enterotoxin. The new enterotoxin also induced morphological changes in L929 cells, whereas the known C. perfringens enterotoxin did not, because L929 cells lacked an appropriate enterotoxin receptor. Although C. perfringens enterotoxin is recognized as the only diarrheagenic toxin responsible for C. perfringens foodborne outbreaks, the results of the present study indicate that C. perfringens isolated from these four outbreaks produced a new type of enterotoxin. PMID:25568432

  10. Clostridium perfringens type A enterotoxin damages the rabbit colon.

    PubMed

    Garcia, Jorge P; Li, Jihong; Shrestha, Archana; Freedman, John C; Beingesser, Juliann; McClane, Bruce A; Uzal, Francisco A

    2014-06-01

    Clostridium perfringens enterotoxin causes the gastrointestinal (GI) symptoms of C. perfringens type A food poisoning and CPE-associated non-food-borne human GI diseases. It is well established that CPE induces fluid accumulation and severe tissue damage in ligated small intestinal loops of rabbits and other animals. However, a previous study had also reported that CPE binds to rabbit colonic cells yet does not significantly affect rabbit colonic loops. To the contrary, the current study determined that treatment with 50 or 100 ?g/ml of CPE causes significant histologic lesions and luminal fluid accumulation in rabbit colonic loops. Interestingly, a CPE-neutralizing monoclonal antibody blocked the development of CPE-induced histologic damage but not luminal fluid accumulation in these loops. Similar luminal fluid accumulation, without significant histologic damage, also occurred after treatment of colonic loops with heat-inactivated CPE, antibody alone, or bovine serum albumin (BSA), indicating that increased osmolarity was causing or contributing to fluid accumulation in CPE-treated colonic loops. Comparative studies revealed the similar development of histologic damage and luminal fluid accumulation in both small intestinal loops and colonic loops after as little as a 1-h treatment with 50 ?g/ml of CPE. Consistent with the CPE sensitivity of the small intestine and colon, Western blotting detected CPE binding and large-complex formation in both organs. In addition, Western blotting demonstrated the presence of the high-affinity CPE receptors claudin-3 and -4 in both organs of rabbits, consistent with the observed toxin binding. Collectively, these results offer support for the possible involvement of the colon in CPE-mediated GI disease. PMID:24643537

  11. Structural Insights into Clostridium perfringens Delta Toxin Pore Formation

    PubMed Central

    Huyet, Jessica; Naylor, Claire E.; Savva, Christos G.; Gibert, Maryse; Popoff, Michel R.; Basak, Ajit K.

    2013-01-01

    Clostridium perfringens Delta toxin is one of the three hemolysin-like proteins produced by C. perfringens type C and possibly type B strains. One of the others, NetB, has been shown to be the major cause of Avian Nectrotic Enteritis, which following the reduction in use of antibiotics as growth promoters, has become an emerging disease of industrial poultry. Delta toxin itself is cytotoxic to the wide range of human and animal macrophages and platelets that present GM2 ganglioside on their membranes. It has sequence similarity with Staphylococcus aureus ?-pore forming toxins and is expected to heptamerize and form pores in the lipid bilayer of host cell membranes. Nevertheless, its exact mode of action remains undetermined. Here we report the 2.4 Å crystal structure of monomeric Delta toxin. The superposition of this structure with the structure of the phospholipid-bound F component of S. aureus leucocidin (LukF) revealed that the glycerol molecules bound to Delta toxin and the phospholipids in LukF are accommodated in the same hydrophobic clefts, corresponding to where the toxin is expected to latch onto the membrane, though the binding sites show significant differences. From structure-based sequence alignment with the known structure of staphylococcal ?-hemolysin, a model of the Delta toxin pore form has been built. Using electron microscopy, we have validated our model and characterized the Delta toxin pore on liposomes. These results highlight both similarities and differences in the mechanism of Delta toxin (and by extension NetB) cytotoxicity from that of the staphylococcal pore-forming toxins. PMID:23805259

  12. Clostridium perfringens Type A Enterotoxin Damages the Rabbit Colon

    PubMed Central

    Garcia, Jorge P.; Li, Jihong; Shrestha, Archana; Freedman, John C.; Beingesser, Juliann; McClane, Bruce A.

    2014-01-01

    Clostridium perfringens enterotoxin causes the gastrointestinal (GI) symptoms of C. perfringens type A food poisoning and CPE-associated non-food-borne human GI diseases. It is well established that CPE induces fluid accumulation and severe tissue damage in ligated small intestinal loops of rabbits and other animals. However, a previous study had also reported that CPE binds to rabbit colonic cells yet does not significantly affect rabbit colonic loops. To the contrary, the current study determined that treatment with 50 or 100 ?g/ml of CPE causes significant histologic lesions and luminal fluid accumulation in rabbit colonic loops. Interestingly, a CPE-neutralizing monoclonal antibody blocked the development of CPE-induced histologic damage but not luminal fluid accumulation in these loops. Similar luminal fluid accumulation, without significant histologic damage, also occurred after treatment of colonic loops with heat-inactivated CPE, antibody alone, or bovine serum albumin (BSA), indicating that increased osmolarity was causing or contributing to fluid accumulation in CPE-treated colonic loops. Comparative studies revealed the similar development of histologic damage and luminal fluid accumulation in both small intestinal loops and colonic loops after as little as a 1-h treatment with 50 ?g/ml of CPE. Consistent with the CPE sensitivity of the small intestine and colon, Western blotting detected CPE binding and large-complex formation in both organs. In addition, Western blotting demonstrated the presence of the high-affinity CPE receptors claudin-3 and -4 in both organs of rabbits, consistent with the observed toxin binding. Collectively, these results offer support for the possible involvement of the colon in CPE-mediated GI disease. PMID:24643537

  13. Quantitative Detection of Clostridium perfringens in the Broiler Fowl Gastrointestinal Tract by Real-Time PCR

    Microsoft Academic Search

    Mark G. Wise; Gregory R. Siragusa

    2005-01-01

    Strains of Clostridium perfringens are a frequent cause of food-borne disease and gas gangrene and are also associated with necrotic enteritis in chickens. To detect and quantify the levels of C. perfringens in the chicken gastrointestinal tract, a quantitative real-time PCR assay utilizing a fluorogenic, hydrolysis-type probe was developed and utilized to assay material retrieved from the broiler chicken cecum

  14. Partial purification and properties of phosphatidylserine synthase from Clostridium perfringens.

    PubMed Central

    Cousminer, J J; Fischl, A S; Carman, G M

    1982-01-01

    The membrane-associated phospholipid biosynthetic enzyme cytidine 5'-diphospho-1,2-diacyl-sn-glycerol:L-serine O-phosphatidyltransferase (phosphatidylserine synthase; EC 2.7.8.8) was partially purified 337-fold from a cell-free extract of the gram-positive pathogenic anaerobe Clostridium perfringens (ATCC 3624). The purification procedure included extraction from the cell envelope with the nonionic detergent Triton X-100, followed by affinity chromatography on cytidine 5'-diphosphate-diacylglycerol-Sepharose. When the partially purified enzyme was subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, two major bands were evident with apparent minimum molecular weights of 39,000 and 31,000. Activity of phosphatidylserine synthase was dependent on the addition of manganese ions (3 mM) and Triton X-100 (2.7 mM) for maximum activity. The rate of catalysis was maximal at 40 degrees C (with rapid thermal inactivation above this temperature), and the pH optimum was 8.5. The apparent Km values for cytidine 5'-diphosphate-diacylglycerol and L-serine were 0.24 and 0.26 mM, respectively. The synthetic (forward) reaction was favored, as indicated by an equilibrium constant of 82, and the energy of activation was found to be 18 kcal/mol (75,362 J/mol). Images PMID:6286597

  15. CDP-diacylglycerol synthase activity in Clostridium perfringens.

    PubMed Central

    Carman, 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 optimum 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 of 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. PMID:6275792

  16. Mechanistic investigations of unsaturated glucuronyl hydrolase from Clostridium perfringens.

    PubMed

    Jongkees, Seino A K; Yoo, Hayoung; Withers, Stephen G

    2014-04-18

    Experiments were carried out to probe the details of the hydration-initiated hydrolysis catalyzed by the Clostridium perfringens unsaturated glucuronyl hydrolase of glycoside hydrolase family 88 in the CAZy classification system. Direct (1)H NMR monitoring of the enzymatic reaction detected no accumulated reaction intermediates in solution, suggesting that rearrangement of the initial hydration product occurs on-enzyme. An attempt at mechanism-based trapping of on-enzyme intermediates using a 1,1-difluoro-substrate was unsuccessful because the probe was too deactivated to be turned over by the enzyme. Kinetic isotope effects arising from deuterium-for-hydrogen substitution at carbons 1 and 4 provide evidence for separate first-irreversible and overall rate-determining steps in the hydration reaction, with two potential mechanisms proposed to explain these results. Based on the positioning of catalytic residues in the enzyme active site, the lack of efficient turnover of a 2-deoxy-2-fluoro-substrate, and several unsuccessful attempts at confirmation of a simpler mechanism involving a covalent glycosyl-enzyme intermediate, the most plausible mechanism is one involving an intermediate bearing an epoxide on carbons 1 and 2. PMID:24573682

  17. Antimicrobial susceptibility of Clostridium perfringens isolated from piglets with or without diarrhea in Brazil

    PubMed Central

    Salvarani, Felipe Masiero; Silveira Silva, Rodrigo Otávio; Pires, Prhiscylla Sadanã; da Costa Cruz Júnior, Eduardo Coulaud; Albefaro, Isabella Silva; de Carvalho Guedes, Roberto Maurício; Faria Lobato, Francisco Carlos

    2012-01-01

    The minimum inhibitory concentration (MIC) was determined for 13 antibiotics against Clostridium perfringens isolated from Brazilian piglets. The collection of isolates was performed in June to October 2010. All isolates were susceptible to amoxicillin and ceftiofur, whereas most were resistant to tetracycline and lincomycin. Avilamycin and narasin were more effective against isolates from non-diarrheic than from diarrheic piglets. The other antimicrobials were less active in need of high concentrations to inhibit the growth of the C. perfringens type A. These results suggest the need for further studies evaluating molecular factors related to the antimicrobial resistance of C. perfringens. PMID:24031924

  18. First isolation of Clostridium perfringens type E from a goat with diarrhea.

    PubMed

    Kim, Ha-Young; Byun, Jae-Won; Roh, In-Soon; Bae, You-Chan; Lee, Myoung-Heon; Kim, Bumseok; Songer, J Glenn; Jung, Byeong Yeal

    2013-08-01

    A 2-day-old goat died suddenly after the onset of severe diarrhea. No specific gross lesions were observed except for a remarkably thin intestinal wall and watery intestinal contents. Histopathological analysis revealed large numbers of Gram-positive bacilli layered upon the intestinal epithelia of the small intestine. Heavy growth of only Clostridium perfringens type E, and no detection of the other enteric pathogens in the small intestine, suggests that C. perfringens type E contributed to the death of this kid. To our knowledge, this is the first isolation of C. perfringens type E from a goat with diarrhea. PMID:23810805

  19. Nitric oxide and lysozyme production as an impact to Clostridium perfringens mastitis.

    PubMed

    Osman, Kamelia M; El-Enbaawy, Mona I; Ezzeldin, Nashwa A; Hussein, Hussein M G

    2010-12-01

    The anaerobic mastitis incidence was used to study the bovine udder response in anaerobic bacterial mastitis caused by the Gram-positive bacterial strain of Clostridium perfringens. Milk samples positive for C. perfringens were assayed for NO and lysozyme. The model produced a strong NO and lysozyme response which correlated positively with the severity and outcome of the disease (subclinical and clinical stages). This study is, to our knowledge, the first to suggest a possible link between NO and lysozyme and bovine mastitis caused by C. perfringens. The results raise the possibility that interfering with NO production during mastitis may help to prevent tissue damage. PMID:19783303

  20. 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

  1. Adhesive properties of Clostridium perfringens to extracellular matrix proteins collagens and fibronectin.

    PubMed

    Hitsumoto, Yasuo; Morita, Naomi; Yamazoe, Ryosuke; Tagomori, Mika; Yamasaki, Tsutomu; Katayama, Seiichi

    2014-02-01

    The adhesive properties of Clostridium perfringens to collagens, gelatin, fibronectin (Fn), Fn-prebound collagens, and Fn-prebound gelatin were investigated. C. perfringens could bind to Fn-prebound collagen type II, type III, and gelatin, but not to gelatin or collagens except for collagen type I directly. Recombinant Fn-binding proteins of C. perfringens, rFbpA and rFbpB, were used to examine Fn-mediated bacterial adherence to collagen type I. In the presence of rFbps, C. perfringens adherence to Fn-prebound collagen type I was inhibited in a dose-dependent manner. Fn was not released from the coated collagen type I by the presence of rFbps, and rFbps did not bind to collagen type I. Thus, the inhibition of C. perfringens binding to Fn-prebound collagen type I by rFbps could not be explained by the removal of Fn from collagen or by the competitive binding of rFbps to collagen. Instead, both rFbps were found to bind to C. perfringens. These results suggest the possibility that rFbps may bind to the putative Fn receptor expressed on C. perfringens and competitively inhibit Fn binding to C. perfringens. PMID:24239649

  2. 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...

  3. Application of Lactobacillus johnsonii expressing phage endolysin for control of Clostridium perfringens.

    PubMed

    Gervasi, T; Lo Curto, R; Minniti, E; Narbad, A; Mayer, M J

    2014-10-01

    Clostridium perfringens is frequently found in food and the environment and produces potent toxins that have a negative impact on both human and animal health and particularly on the poultry industry. Lactobacillus johnsonii FI9785, isolated from the chicken gastrointestinal tract, has been demonstrated to exclude Cl. perfringens in poultry. We have investigated the interaction of wild-type Lact. johnsonii FI9785 or an engineered strain expressing a cell wall-hydrolysing endolysin with Cl. perfringens in vitro, using a batch culture designed to simulate human gastrointestinal tract conditions. Co-culture experiments indicated that acid production by Lact. johnsonii is important in pathogen control. The co-culture of the endolysin-secreting Lact. johnsonii with Cl. perfringens showed that the engineered strain had the potential to control the pathogen, but the ability to reduce Cl. perfringens numbers was not consistent. Results obtained indicate that survival of high numbers of Lact. johnsonii will be essential for effective pathogen control. Significance and impact of the study: The bacterium Lactobacillus johnsonii FI9785 reduces numbers of the pathogen Clostridium perfringens in vitro. Biocontrol was improved by engineering the strain to produce and export a cell wall-hydrolysing endolysin, but good survival of the producer strain is essential. The production of bacteriophage endolysins by commensal bacteria has the potential to improve competitive exclusion of pathogens in the gastrointestinal tract. PMID:24961379

  4. ESTIMATATION OF GROWTH OF CLOSTRIDIUM PERFRINGENS IN COOKED BEEF UNDER FLUCTUATING TEMPERATURE CONDITIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new concept for estimating the bacterial growth under temperature fluctuations was hypothesized and validated using Clostridium perfringens as a test organism. This new methodology was based on the Gompertz models to calculate the equivalent growth times under different temperatures, and estimate...

  5. Control of Clostridium perfringens spores by plant-derived antimicrobials during cooling of cooked ground beef

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens spore germination and outgrowth by carvacrol, cinnamaldehyde, thymol, oregano oil and two green tea extracts with low (green tea leaf powder (GTL); 141 mg of total catechins/g of green tea extract) and high (green tea leaf extract (GTE); 697 mg of total catechin...

  6. 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...

  7. Complete genome sequence of the podoviral bacteriophage CP24R virulent for Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacteriophage 'CP24R was isolated from raw sewage of a waste treatment plant and lytic activity was observed against a type C Clostridium perfringens isolate. Electron microscopy revealed a small virion (44nm diameter icosahedral capsid) with a short, non-contractile tail, indicative of the family ...

  8. 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...

  9. 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

  10. BEC, a Novel Enterotoxin of Clostridium perfringens Found in Human Clinical Isolates from Acute Gastroenteritis Outbreaks

    PubMed Central

    Yonogi, Shinya; Matsuda, Shigeaki; Kawai, Takao; Yoda, Tomoko; Harada, Tetsuya; Kumeda, Yuko; Gotoh, Kazuyoshi; Hiyoshi, Hirotaka; Nakamura, Shota; Kodama, Toshio

    2014-01-01

    Clostridium perfringens is a causative agent of food-borne gastroenteritis for which C. perfringens enterotoxin (CPE) has been considered an essential factor. Recently, we experienced two outbreaks of food-borne gastroenteritis in which non-CPE producers of C. perfringens were strongly suspected to be the cause. Here, we report a novel enterotoxin produced by C. perfringens isolates, BEC (binary enterotoxin of C. perfringens). Culture supernatants of the C. perfringens strains showed fluid-accumulating activity in rabbit ileal loop and suckling mouse assays. Purification of the enterotoxic substance in the supernatants and high-throughput sequencing of genomic DNA of the strains revealed BEC, composed of BECa and BECb. BECa and BECb displayed limited amino acid sequence similarity to other binary toxin family members, such as the C. perfringens iota toxin. The becAB genes were located on 54.5-kb pCP13-like plasmids. Recombinant BECb (rBECb) alone had fluid-accumulating activity in the suckling mouse assay. Although rBECa alone did not show enterotoxic activity, rBECa enhanced the enterotoxicity of rBECb when simultaneously administered in suckling mice. The entertoxicity of the mutant in which the becB gene was disrupted was dramatically decreased compared to that of the parental strain. rBECa showed an ADP-ribosylating activity on purified actin. Although we have not directly evaluated whether BECb delivers BECa into cells, rounding of Vero cells occurred only when cells were treated with both rBECa and rBECb. These results suggest that BEC is a novel enterotoxin of C. perfringens distinct from CPE, and that BEC-producing C. perfringens strains can be causative agents of acute gastroenteritis in humans. Additionally, the presence of becAB on nearly identical plasmids in distinct lineages of C. perfringens isolates suggests the involvement of horizontal gene transfer in the acquisition of the toxin genes. PMID:24664508

  11. Freshwater Suspended Sediments and Sewage Are Reservoirs for Enterotoxin-Positive Clostridium perfringens?

    PubMed Central

    Mueller-Spitz, Sabrina R.; Stewart, Lisa B.; Klump, J. Val; McLellan, Sandra L.

    2010-01-01

    The release of fecal pollution into surface waters may create environmental reservoirs of feces-derived microorganisms, including pathogens. Clostridium perfringens is a commonly used fecal indicator that represents a human pathogen. The pathogenicity of this bacterium is associated with its expression of multiple toxins; however, the prevalence of C. perfringens with various toxin genes in aquatic environments is not well characterized. In this study, C. perfringens spores were used to measure the distribution of fecal pollution associated with suspended sediments in the nearshore waters of Lake Michigan. Particle-associated C. perfringens levels were greatest adjacent to the Milwaukee harbor and diminished in the nearshore waters. Species-specific PCR and toxin gene profiles identified 174 isolates collected from the suspended sediments, surface water, and sewage influent as C. perfringens type A. Regardless of the isolation source, the beta2 and enterotoxin genes were common among isolates. The suspended sediments yielded the highest frequency of cpe-carrying C. perfringens (61%) compared to sewage (38%). Gene arrangement of enterotoxin was investigated using PCR to target known insertion sequences associated with this gene. Amplification products were detected in only 9 of 90 strains, which suggests there is greater variability in cpe gene arrangement than previously described. This work presents evidence that freshwater suspended sediments and sewage influent are reservoirs for potentially pathogenic cpe-carrying C. perfringens spores. PMID:20581181

  12. 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 ...

  13. Comparative genomics of four closely related Clostridium perfringens bacteriophages reveals variable rates of evolution within a core genome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Biotechnological uses of bacteriophage gene products as alternatives to conventional antibiotics will require a thorough understanding of their genomic context. We sequenced and analyzed the genomes of four closely related phages isolated from Clostridium perfringens, an important agricu...

  14. 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.

  15. SleC Is Essential for Cortex Peptidoglycan Hydrolysis during Germination of Spores of the Pathogenic Bacterium Clostridium perfringens

    Microsoft Academic Search

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

    2009-01-01

    Clostridial spore germination requires degradation of the spore's peptidoglycan (PG) cortex by cortex-lytic enzymes (CLEs), and two Clostridium perfringens CLEs, SleC and SleM, degrade cortex PG in vitro. We now find that only SleC is essential for cortex hydrolysis and viability of C. perfringens spores. C. perfringens sleC spores did not germinate completely with nutrients, KCl, or a 1:1 chelate

  16. BACTERIOCIN E1073 PRODUCED BY ENTEROCOCCUS FAECIUM LWP1073 IS EFFECTIVE FOR TREATING COMMENSAL CLOSTRIDIUM PERFRINGENS INFECTION IN BROILERS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enterotoxin-producing Clostridium perfringens type A bacteria occupy a significant place in the etiological structure of food-borne infections in humans. One potential approach to minimize infections associated with food-borne pathogens is to control the carriage of C. perfringens in broilers. For ...

  17. 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, ...

  18. The molecular-genetic analysis of Clostridium perfringens strains isolated from broilers on farms in Central Russia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of the research was to perform phenotypic and molecular-genetic typing of Clostridium perfringens strains commonly spread on poultry farms in Central Russia. Samples of homogenized iliac and cecal contents from 760 broilers were assayed and 325 C. perfringens strains (42.8 %) were isol...

  19. GROWTH OF CLOSTRIDIUM PERFRINGENS FROM SPORE INOCULA AT TEMPERATURES APPLICABLE TO COOLING OF COOKED MEAT AND POULTRY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of the germination and outgrowth of Clostridium perfringens by chitosan during the abusive chilling of beef and turkey was evaluated. Chilling of cooked beef and turkey from 54.4 to 7.2 degrees C resulted in C. perfringens population increases of > 5 log10 cfu/g during 18 h exponential...

  20. ANALYSIS OF CORE HOUSEKEEPING AND VIRULENCE GENES REVEALS CRYPTIC LINEAGES OF CLOSTRIDIUM PERFRINGENS THAT ARE ASSOCIATED WITH DISTINCT DISEASE PRESENTATIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium perfringens is an important human and animal pathogen that causes a number of different diseases that vary in their etiology and severity. Currently, the identification of C. perfringens strains is based solely upon the presence and absence of particular toxin genes. Unfortunately, thi...

  1. Toxinotyping of Clostridium perfringens fecal isolates of reintroduced Père David's deer (Elaphurus davidianus) in China.

    PubMed

    Qiu, Huiling; Chen, Fu; Leng, Xinyan; Fei, Rongmei; Wang, Libo

    2014-10-01

    Clostridium perfringens is an important pathogen causing sudden death syndrome, necrotic enteritis, and gas gangrene in ruminants, especially some deer species. Père David's deer (Elaphurus davidianus) is one of the world's rare species and is an endangered and protected species in China. Some Père David's deer in the Chinese Shishou Père David's Deer Preserve died due to C. perfringens infection. We investigated the toxin types and C. perfringens enterotoxin-positive (cpe(+)) strains of isolated C. perfringens in Père David's deer in China. We collected 155 fecal samples from the Beijing Nanhaizi Père David's Deer Park and the Jiangsu Dafeng Père David's Deer National Nature Reserve between July 2010 and July 2011. Bacteria isolated using blood agar and mannitol agar plates were identified by Gram staining and nested PCR for 16S rRNA. We isolated C. perfringens from 41 fecal samples and used PCR amplification of five toxin genes to identify the toxinotypes and the cpe(+) strains of C. perfringens. Twenty-one isolates were type A, 15 were type E, and five were type D. Fifteen isolates were cpe(+) strains, including eight that were type A and seven that were type E. PMID:25050802

  2. Use of ?-irradiation to reduce Clostridium perfringens on ready-to-eat bovine tripe.

    PubMed

    Parry-Hanson, Angela; Hall, Alan; Minnaar, Amanda; Buys, Elna M

    2008-03-01

    The aim of this study was to determine the effect of gamma irradiation at a target dose of 9kGy and storage at 5 and 15°C on the safety of ready-to-eat (RTE) tripe with respect to Clostridium perfringens count (CC) and aerobic plate count (APC), and to determine the effect of boiling (1h) and irradiation (9kGy) on Cl. perfringens ATCC 13124 spore structure. Irradiation significantly reduced APC stored at 5 and 15°C for 7 days. However, 0kGy control samples increased in their APC to >7log(10) cfu/g throughout 7 days of storage. Irradiation eliminated the inoculated Cl. perfringens ATCC 13124 spores on RTE tripe throughout storage at 5 and 15°C. Transmission electron microscopy of Cl. perfringens ATCC 13124 spores showed that boiling caused a reduction in spore material, irradiation caused elongation of the Cl. perfringens ATCC 13124 spores, and boiling in combination with gamma irradiation caused loss of spore material. Therefore, irradiation at 9kGy, together with storage at 5°C, can assure the microbiological safety of RTE bovine tripe, with respect to Cl. perfringens spores for at least 7 days at 5 and 15°C. PMID:22062270

  3. The inhibitory effects of sorbate and benzoate against Clostridium perfringens type A isolates.

    PubMed

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

    2015-06-01

    This study evaluated the inhibitory effects of sorbate and benzoate against Clostridium perfringens type A food poisoning (FP) and non-food-borne (NFB) disease isolates. No significant inhibition of germination of spores of both FP and NFB isolates was observed in rich medium (pH 7.0) supplemented with permissive level of sodium sorbate (0.3% ? 0.13 mM undissociated sorbic acid) or sodium benzoate (0.1% ? 0.01 mM undissociated benzoic acid) used in foods. However, these levels of sorbate and benzoate effectively arrested outgrowth of germinated C. perfringens spores in rich medium. Lowering the pH of the medium increases the inhibitory effects of sorbate and benzoate against germination of spores of NFB isolates, and outgrowth of spores of both FP and NFB isolates. Furthermore, sorbate and benzoate inhibited vegetative growth of C. perfringens isolates. However, the permissible levels of these organic salts could not control the growth of C. perfringens spores in chicken meat stored under extremely abusive conditions. In summary, although sorbate and benzoate showed inhibitory activities against C. perfringens in the rich medium, no such effect was observed in cooked chicken meat. Therefore, caution should be taken when applying these organic salts into meat products to reduce or eliminate C. perfringens spores. PMID:25790996

  4. 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

  5. Optimized necrotic enteritis model producing clinical and subclinical infection of Clostridium perfringens in broiler chickens.

    PubMed

    Wu, Shu-Biao; Rodgers, Nicholas; Choct, Mingan

    2010-09-01

    In this study we assessed the roles of Eimeria infection and dietary manipulation (feeding a diet with a high level of fishmeal) in an Australian necrotic enteritis (NE) challenge model in broiler chickens. An experiment was designed to test the hypothesis that Eimeria infection and dietary manipulation, i.e., inclusion of fishmeal in the diet, are necessary to induce NE experimentally. The results showed that the combination of Eimeria administration and fishmeal feeding had a significant effect on induction of clinical and subclinical Clostridium perfringens infection. The majority of the mortality that occurred during the second week of the trial was due to an NE outbreak following the C. perfringens challenge. The mortality rate of the birds was 12.00% for the high-fishmeal (HFM; 500 g/kg) group and 9.33% for the low-fishmeal (LFM; 250 g/kg) group when the birds were subjected to C. perfringens and Eimeria. Fishmeal alone did not induce significant mortality in birds challenged only with C. perfringens but showed a significantly higher C. perfringens count than the non-fishmeal (NFM) control group. Eimeria administration had a significant effect on NE-related mortality but did not have an effect on the C. perfringens count. In accordance with the time course of bird mortality, it can be determined that of the 3 successive days of oral gavage with C. perfringens, the first inoculation was essential for inducing NE, but the third had no additional effect on NE-related mortality. Also, reducing the fishmeal level from 500 to 250 g/kg had no negative impact on the reproducibility of the model. It may be concluded that NE can be consistently induced under experimental conditions by feeding broilers a diet containing 250 g/kg fishmeal, using a single inoculation with low numbers of Eimeria, administering one or two oral C. perfringens inoculations, and maintaining appropriate ambient temperatures and diets. PMID:20945788

  6. 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

  7. Necrotizing enterocolitis of the neonate with Clostridium perfringens: diagnosis, clinical course, and role of alpha toxin

    Microsoft Academic Search

    Elke Dittmar; Peter Beyer; Doris Fischer; Volker Schäfer; Heike Schoepe; Karl Bauer; Rolf Schlösser

    2008-01-01

    The severity of the clinical course in necrotizing enterocolitis (NEC) associated with Clostridium perfringens (Cp) may support the hypothesis of a specific disease. We conducted a case control study of infants diagnosed with NEC, who\\u000a underwent surgical treatment over a 7-year period. Patient histories examined characteristics of the infants, bacterial infection\\u000a as well as NEC’s severity, antibiotic treatment, and clinical

  8. Role of DNase in recovery of plasmid DNA from Clostridium perfringens.

    PubMed Central

    Blaschek, H P; Klacik, M A

    1984-01-01

    Recovery of plasmid DNA from Clostridium perfringens 10543A and 3626B cleared lysates was significantly improved by the addition of 0.2% (vol/vol) diethylpyrocarbonate (DEP) before protoplast disruption in the cleared lysate protocol. Three previously undetected, large-molecular-mass plasmids (45.2, 51.9, and 68.2 megadaltons) were isolated from modified DEP-treated cleared lysates of C. perfringens 3626B. Two plasmids (9.4 and 30 megadaltons) were recovered from C. perfringens 10543A modified DEP-treated cleared lysates which previously required dye-buoyant density gradient centrifugation for visualization on agarose gels. Unsuccessful attempts to isolate plasmid DNA from Brij 58 cleared lysates of extracellular DNase-negative mutants of C. perfringens suggested the deleterious DNase activity was not extracellular. Cellular localization studies indicated that the cell wall-compartmentalized cell fraction contained 72.2% of the total DNase activity, whereas the extracellular and intracellular fractions demonstrated much less (26.8 and 1.0%, respectively). Cleared lysates prepared with DEP demonstrated much less DNase activity than cleared lysates prepared without DEP. The variable and irreproducible recovery of plasmid DNA from C. perfringens cleared lysates was attributed to cell wall-compartmentalized DNase. Images PMID:6089664

  9. Membrane vesicles of Clostridium perfringens type A strains induce innate and adaptive immunity.

    PubMed

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

    2014-05-01

    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

  10. A low-toxic site-directed mutant of Clostridium perfringens ?-toxin as a potential candidate vaccine against enterotoxemia.

    PubMed

    Li, Qing; Xin, Wenwen; Gao, Shan; Kang, Lin; Wang, Jinglin

    2013-11-01

    Clostridium perfringens epsilon toxin (ETX), one of the most potent toxins known, is a potential biological weapon; therefore, the development of an effective vaccine is important for preventing intoxication or disease by ETX. In this study, genetically detoxified epsilon toxin mutants were developed as candidate vaccines. We used site-directed mutagenesis to mutate the essential amino acid residues (His106, Ser111 and Phe199). Six site-directed mutants of ETX (mETX (H106P) , mETX (S111H) , mETX (S111Y) , mETX (F199H) , mETX (F199E) , mETX (S111YF199E) ) were generated and then expressed in Escherichia coli. Both mETX (F199E) and mETX (H106P) with low or non-cytotoxicity that retained their immunogenicity were selected to immunize mice 3 times, and the mouse survival data were recorded after challenging with recombinant wild-type ETX. mETX (F199E) induces the same protection as mETX (H106P) , which was reported previously as a promising toxin mutant for vaccine, and both of them could protect immunized mice against a 100× LD?? dose of active wild-type recombinant ETX. This work showed that mETX (F199E) is another promising candidate vaccine against enterotoxemia and other diseases caused by ETX. PMID:23835363

  11. A low-toxic site-directed mutant of Clostridium perfringens ?-toxin as a potential candidate vaccine against enterotoxemia

    PubMed Central

    Li, Qing; Xin, Wenwen; Gao, Shan; Kang, Lin; Wang, Jinglin

    2013-01-01

    Clostridium perfringens epsilon toxin (ETX), one of the most potent toxins known, is a potential biological weapon; therefore, the development of an effective vaccine is important for preventing intoxication or disease by ETX. In this study, genetically detoxified epsilon toxin mutants were developed as candidate vaccines. We used site-directed mutagenesis to mutate the essential amino acid residues (His106, Ser111 and Phe199). Six site-directed mutants of ETX (mETXH106P, mETXS111H, mETXS111Y, mETXF199H, mETXF199E, mETXS111YF199E) were generated and then expressed in Escherichia coli. Both mETXF199E and mETXH106P with low or non-cytotoxicity that retained their immunogenicity were selected to immunize mice 3 times, and the mouse survival data were recorded after challenging with recombinant wild-type ETX. mETXF199E induces the same protection as mETXH106P, which was reported previously as a promising toxin mutant for vaccine, and both of them could protect immunized mice against a 100× LD50 dose of active wild-type recombinant ETX. This work showed that mETXF199E is another promising candidate vaccine against enterotoxemia and other diseases caused by ETX. PMID:23835363

  12. 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

  13. 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

  14. 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

  15. 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

  16. [Studies of necrotizing enteritis of suckling piglets (Clostridium perfringens type C enterotoxemia) in industrialized sow breeding units. 4. Epizootiology].

    PubMed

    Köhler, B; Zabke, J; Sondermann, R; Pulst, H; Rummler, H J

    1979-01-01

    Necrotising enteritis had been the cause of death of 4.9 per cent in 5,177 nursed piglets, which was established by pathological examination. The number of piglets, in that context, which had come from industrialised sow breeding units was equivalent to 92 per cent. The nursed piglet held the third position, next to smaller ruminants (19.4 per cent) and fowl (6.0 per cent), with regard to the occurrence of Clostridium perfringens enterotoxemia or necrotising enteritis in 112,218 animals which were pathologically examined after death. Necrotising enteritis so far has been rare in the GDR. No regional accumulation has been observed. Several outbreaks on industrialised sow breeding units actually remained stationary. The occurrence of the disease may be favoured by a number of factors which are conducive to accumulation of Clostridium perfringens Type C in a given stock. Group keeping of pregnant sows, simultaneous farrowing of larger groups of sows, group treatment of nursed piglets, using neomycin, chloramphenicol, oxytetracycline, and other antibiotics to which Clostridium perfringens is primarily resistant or has acquired resistance in the course of time are some of those contributive factors. Transmission of Clostridium perfringens Type C through feedstuff is possible, though it would lead to a real outbreak only by high intensity of the contamination, and it played a minor role in proliferation of the disease. 3479 Clostridium perfringens strains were isolated from 9,481 animals, both clinically intact and after death, with 30 species being included. Type classification revealed 2454 strains of Type A (70 per cent), 204 of Type D (5.88 per cent), 164 of Type C (four per cent), and 48 of Type B (1.34 per cent). There were 688 atoxic strains (17 per cent). Swine is the major carrier of Clostridium perfringens Type C, with 87 per cent of all Clostridium perfringens Type C strains having been isolated from swine. Swine was followed by fowl (four per cent), sheep (four per cent), cattle, rabbit, and dog (1.27 per cent each). Clostridium perfringens Type C was obtained from the faeces of clinically intact sows in seven instances, including two cases with sows (0.46 per cent) from farms with no previous record of necrotising enteritis. PMID:232840

  17. Comparative Proteomic Analysis of Extracellular Proteins of Clostridium perfringens Type A and Type C Strains? †

    PubMed Central

    Sengupta, Nabonita; Alam, Syed Imteyaz; Kumar, Bhoj; Kumar, Ravi Bhushan; Gautam, Vandana; Kumar, Subodh; Singh, Lokendra

    2010-01-01

    Clostridium perfringens is a medically important clostridial pathogen and an etiological agent causing several diseases in humans and animals. C. perfringens and its toxins have been listed as potential biological and toxin warfare (BTW) agents; thus, efforts to develop strategies for detection and protection are warranted. Forty-eight extracellular proteins of C. perfringens type A and type C strains have been identified here using a 2-dimensional gel electrophoresis-mass spectrometry (2-DE-MS) technique. The SagA protein, the DnaK-type molecular chaperone hsp70, endo-beta-N-acetylglucosaminidase, and hypothetical protein CPF_0656 were among the most abundant proteins secreted by C. perfringens ATCC 13124. The antigenic component of the exoproteome of this strain has also been identified. Most of the extracellular proteins were predicted to be involved in carbohydrate transport and metabolism (16%) or cell envelope biogenesis or to be outer surface protein constituents (13%). More than 50% of the proteins were predictably secreted by either classical or nonclassical pathways. LipoP and TMHMM indicated that nine proteins were extracytoplasmic but cell associated. Immunization with recombinant ornithine carbamoyltransferase (cOTC) clearly resulted in protection against a direct challenge with C. perfringens organisms. A significant rise in IgG titers in response to recombinant cOTC was observed in mice, and IgG2a titers predominated over IgG1 titers (IgG2a/IgG1 ratio, 2). The proliferation of spleen lymphocytes in cOTC-immunized animals suggested a cellular immune response. There were significant increases in the levels of gamma interferon (IFN-?) and interleukin 2 (IL-2), suggesting a Th1 type immune response. PMID:20605988

  18. 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

  19. Correlations Between Virulence and Other Characters of Clostridium perfringens Type A

    PubMed Central

    Forget, A.; Paquette, G.; Roy, A.; Fredette, V.

    1969-01-01

    The correlation analysis which has already proved its value in ecology has not yet been applied to the determination of virulence indicators. Its application to a group of Clostridium perfringens type A strains has brought out some characters that may be considered as virulence indicators. This study suggests that the toxicity of the culture supernatant fluids for mice is significantly correlated with the virulence for mice and guinea pigs. A significant correlation was found between the virulence of the fluid cultures for mice or guinea pigs and the coagulation of milk, production of gas (in deep agar), hydrogen sulfide production, and fermentation of glucose, sucrose, maltose, and levulose. PMID:4312926

  20. Benthic distribution of sewage sludge indicated by Clostridium perfringens at a deep-ocean dump site

    SciTech Connect

    Hill, R.T.; Anikis, M.S.; Colwell, R.R. (Univ. of Maryland, Baltimore (United States)); Knight, I.T. (James Madison Univ., Harrisonburg, VA (United States))

    1993-01-01

    Since 1986, sewage sludge from New York and northern New Jersey has been dumped 196 km off the coast of New Jersey at the Deep Water Municipal Sewage Sludge Disposal Site. This study determines the distribution of sludge contamination of the benthic environment in the area, by using Clostridium perfringens as an indicator. The counts of C. perfringens confirm a previous report that sewage sludge is reaching the ocean floor at the disposal site as a result of the sludge dumping. C. perfringes counts within the dump site and to the south and west of the dump site are considerably elevated compared to counts east of the site. The distribution pattern of C. perfringes is broadly consistent with the estimates of the sea floor area impacted in the most recent computer model. However, the area of maximum desposition of sludge may be slightly further north than predicted. Use of C. perfringens has proven to be an efficient and reliable method for tracing sewage contamination of deep ocean sediments. 18 refs., 4 figs., 3 tabs.

  1. Identification of a two-component signal transduction system that regulates maltose genes in Clostridium perfringens.

    PubMed

    Hiscox, Thomas J; Ohtani, Kaori; Shimizu, Tohru; Cheung, Jackie K; Rood, Julian I

    2014-12-01

    Clostridium perfringens is a Gram-positive rod that is widely distributed in nature and is the etiological agent of several human and animal diseases. The complete genome sequence of C. perfringens strain 13 has been determined and multiple two-component signal transduction systems identified. One of these systems, designated here as the MalNO system, was analyzed in this study. Microarray analysis was used to carry out functional analysis of a malO mutant. The results, which were confirmed by quantitative reverse-transcriptase PCR, indicated that genes putatively involved in the uptake and metabolism of maltose were up-regulated in the malO mutant. These effects were reversed by complementation with the wild-type malO gene. Growth of these isogenic strains in medium with and without maltose showed that the malO mutant recovered more quickly from maltose deprivation when compared to the wild-type and complemented strains, leading to the conclusion that the MalNO system regulates maltose utilization in C. perfringens. It is postulated that this regulatory network may allow this soil bacterium and opportunistic pathogen to respond to environmental conditions where there are higher concentrations of maltose or maltodextrins, such as in the presence of decaying plant material in rich soil. PMID:25152227

  2. Growth conditions of clostridium perfringens type B for production of toxins used to obtain veterinary vaccines.

    PubMed

    Viana Brandi, Igor; Domenici Mozzer, Otto; Jorge, Edson Vander; Vieira Passos, Frederico Jose; Lopes Passos, Flavia Maria; Cangussu, Alex Sander Rodrigues; Macedo Sobrinho, Eliane

    2014-09-01

    The diseases caused for Clostridium perfringens are generically called enterotoxemias because toxins produced in the intestine may be absorbed into the general circulation. C. perfringens type B, grown in batch fermentation, produced toxins used to obtain veterinary vaccines. Glucose in concentrations of 1.4-111.1 mM was used to define the culture medium. The minimum concentration for a satisfactory production of vaccines against clostridial diseases was 55.6 mM. Best results were brought forth by meat and casein peptones, both in the concentration 5.0 g l(-1) in combination with glucose and a culture pH maintained at 6.5 throughout the fermentation process. The production of lactic, acetic and propionic organic acids was observed. Ethanol was the metabolite produced in the highest concentration when cultures maintained steady pH of 6.5 with exception of cultures with initial glucose concentration of 1.4 mM, where the highest production was of propionic acid. Maximal cell concentration and the highest toxin title concomitantly low yield coefficient to organic acids and ethanol were obtained using basal medium containing 111.1 mM glucose under a controlled pH culture (pH) 6.5 in batch fermentations of C. perfringens type B. These data contribute to improve process for industrial toxin production allowing better condition to produce a toxoid vaccine. PMID:24573216

  3. INFLUENCE OF NAC1 CONTENT AND COOLING RATE ON OUTGROWTH OF CLOSTRIDIUM PERFRINGENS SPORES IN COOKED HAM AND BEEF

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of NaCl concentration and cooling rate on the ability of Clostridium perfringens to grow from spore inocula was studied using a process that simulates the industrial cooking and cooling of smoked boneless ham and beef roast. Sufficient NaCl was added to 2 different ground commercially-ob...

  4. INHIBITION OF GERMINATION AND OUTGROWTH OF CLOSTRIDIUM PERFRINGENS SPORES BY LACTIC ACID SALTS DURING COOLING OF COOKED GROUND TURKEY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inhibition of Clostridium perfringens spore germination and outgrowth by lactic acid salts during exponential cooling of cooked ground turkey products was evaluated. Injected turkey containing either calcium lactate, potassium lactate, or sodium lactate (1.0, 2.0, 3.0 or 4.8% w/w) along with a cont...

  5. 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...

  6. Draft Genome Sequence of Clostridium perfringens Strain JJC, a Highly Efficient Hydrogen Producer Isolated from Landfill Leachate Sludge

    PubMed Central

    Wong, Y. M.; Gan, H. M.; Austin, C. M.

    2014-01-01

    Clostridium perfringens strain JJC is an effective biohydrogen and biochemical producer that was isolated from landfill leachate sludge. Here, we present the assembly and annotation of its genome, which may provide further insights into the gene interactions involved in efficient biohydrogen production. PMID:24604637

  7. 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, ...

  8. Changing a Single Amino Acid in Clostridium perfringens ?-Toxin Affects the Efficiency of Heterologous Secretion by Bacillus subtilis

    Microsoft Academic Search

    Reindert Nijland; René Heerlien; Leendert W. Hamoen; Oscar P. Kuipers

    2007-01-01

    Achieving efficient heterologous protein production and secretion by Bacillus subtilis is an attractive prospect, although often disappointingly low yields are reached. The expression of detoxified Clostridium perfringens ?-toxin (?-toxoid) is exemplary for this. Although ?-toxin can be efficiently expressed and secreted by Bacillus subtilis, the genetically detoxified, and industrially interesting, ?-toxoid variant is difficult to obtain in high amounts. To

  9. 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...

  10. 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

  11. Identification of Clostridium Species and DNA Fingerprinting of Clostridium perfringens by Amplified Fragment Length Polymorphism Analysis

    Microsoft Academic Search

    Riikka Keto-Timonen; Annamari Heikinheimo; Erkki Eerola; Hannu Korkeala

    2006-01-01

    An amplified fragment length polymorphism (AFLP) method was applied to 129 strains representing 24 different Clostridium species, with special emphasis on pathogenic clostridia of medical or veterinary interest, to assess the potential of AFLP for identification of clostridia. In addition, the ability of the same AFLP protocol to type clostridia at the strain level was assessed by focusing on Clostridium

  12. Epidemiological and pathobiological profiles of Clostridium perfringens infections: review of consecutive series of 33 cases over a 13-year period

    PubMed Central

    Shindo, Yuji; Dobashi, Yoh; Sakai, Toshiyasu; Monma, Chie; Miyatani, Hiroyuki; Yoshida, Yukio

    2015-01-01

    Background: Although Clostridium perfringens (C. perfringens) is well known as the causative agent of several forms of enteric disease, precise epidemiological and pathobiological aspects are still unknown. Methods: We retrospectively reviewed the culture results of samples collected in our hospital from 2001 through 2013. In addition, for the detection and toxinogenic typing of C. perfringens, polymerase-chain-reaction amplification (PCR)-based rapid analysis was performed in 6 cases using DNA extracted from paraffin-embedded tissues. Results: A total of 35 samples from 33 cases were positive for C. perfringens, representing an incidence of 0.017% (35/205, 114). Among 33 patients, 21 patients manifested sepsis and 7 patients had bacteremia. One of the septic cases was complicated by fatal intravascular hemolysis and thus, the prevalence was estimated at 3.0% among C. perfringens infections (1/33). The direct causative disease or state for C. perfringens infection was identified in 18 patients: surgery or intervention for cancers, 8 patients; chemotherapy for cancer, 2 patients; surgery or intervention for non-neoplastic disease, 6 patients; liver cirrhosis, 3 patients, etc. PCR-based toxinogenic typing of C. perfringens detected the alpha-toxin gene only in tissue from a patient who died of massive hemolysis; none of the toxin genes could be amplified in the other 5 cases examined. Conclusions: The prevalence of overt C. perfringens infection is low, but upon detection, infected patients should be carefully monitored for fatal acute hemolysis caused by type A C. perfringens. Furthermore, PCR-based rapid detection of C. perfringens and toxinogenic typing by archival pathological material is applicable as a diagnostic tool. PMID:25755747

  13. Synergistic effects of Clostridium perfringens enterotoxin and beta toxin in rabbit small intestinal loops.

    PubMed

    Ma, Menglin; Gurjar, Abhijit; Theoret, James R; Garcia, Jorge P; Beingesser, Juliann; Freedman, John C; Fisher, Derek J; McClane, Bruce A; Uzal, Francisco A

    2014-07-01

    The ability of Clostridium perfringens type C to cause human enteritis necroticans (EN) is attributed to beta toxin (CPB). However, many EN strains also express C. perfringens enterotoxin (CPE), suggesting that CPE could be another contributor to EN. Supporting this possibility, lysate supernatants from modified Duncan-Strong sporulation (MDS) medium cultures of three CPE-positive type C EN strains caused enteropathogenic effects in rabbit small intestinal loops, which is significant since CPE is produced only during sporulation and since C. perfringens can sporulate in the intestines. Consequently, CPE and CPB contributions to the enteropathogenic effects of MDS lysate supernatants of CPE-positive type C EN strain CN3758 were evaluated using isogenic cpb and cpe null mutants. While supernatants of wild-type CN3758 MDS lysates induced significant hemorrhagic lesions and luminal fluid accumulation, MDS lysate supernatants of the cpb and cpe mutants caused neither significant damage nor fluid accumulation. This attenuation was attributable to inactivating these toxin genes since complementing the cpe mutant or reversing the cpb mutation restored the enteropathogenic effects of MDS lysate supernatants. Confirming that both CPB and CPE are needed for the enteropathogenic effects of CN3758 MDS lysate supernatants, purified CPB and CPE at the same concentrations found in CN3758 MDS lysates also acted together synergistically in rabbit small intestinal loops; however, only higher doses of either purified toxin independently caused enteropathogenic effects. These findings provide the first evidence for potential synergistic toxin interactions during C. perfringens intestinal infections and support a possible role for CPE, as well as CPB, in some EN cases. PMID:24778117

  14. Association between avian necrotic enteritis and Clostridium perfringens strains expressing NetB toxin

    PubMed Central

    Keyburn, Anthony L.; Yan, Xu-Xia; Bannam, Trudi L.; Van Immerseel, Filip; Rood, Julian I.; Moore, Robert J.

    2009-01-01

    A novel toxin, NetB, has recently been identified in virulent avian Clostridium perfringens isolates and shown to be an essential virulence factor in a clinical necrotic enteritis isolate. To assess whether NetB is more generally associated with avian necrotic enteritis isolates we have screened a range of C. perfringens strains from geographically diverse locations for both the presence and expression of the netB gene. Forty-four isolates were derived from necrotic enteritis disease cases from Australia, Belgium, Denmark and Canada and 55 isolates from healthy chickens from Australia and Belgium. The majority of strains isolated from necrotic enteritis-affected birds were netB positive (70%) and there was an absolute correlation between the presence of netB and in vitro expression of the NetB protein. Only two of the C. perfringens isolates from healthy chickens carried netB. Sequencing of the netB gene from 23 positive isolates showed that NetB is highly conserved, with only one predicted amino acid (A168T) difference, in six isolates, compared to the published sequence. This change did not alter the in vitro activity of the NetB toxin. The gene encoding the recently discovered TpeL toxin was also screened using PCR and only found in a small proportion of NetB-positive isolates from diseased birds. A selection of NetB-negative isolates, originating from diseased birds, was unable to cause disease in a necrotic enteritis induction model. This study provides further evidence that NetB is important in pathogenesis and advances our current understanding of C. perfringens virulence factors in avian necrotic enteritis. PMID:19931005

  15. 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

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

    PubMed

    Juneja, Vijay K; Baker, David A; Thippareddi, H; Snyder, O Peter; Mohr, Tim B

    2013-01-01

    The ability of Clostridium perfringens to germinate and grow in acidified ground beef as well as in 10 commercially prepared acidified beef, pork, and poultry products was assessed. The pH of ground beef was adjusted with organic vinegar to achieve various pH values between 5.0 and 5.6; the pH of the commercial products ranged from 4.74 to 6.35. Products were inoculated with a three-strain cocktail of C. perfringens spores to achieve ca. 2-log (low) or 4-log (high) inoculum levels, vacuum packaged, and cooled exponentially from 54.4 to 7.2°C for 6, 9, 12, 15, 18, or 21 h to simulate abusive cooling; the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) recommends a cooling time of 6.5 h. Total germinated C. perfringens populations were determined after plating on tryptose-sulfite-cycloserine agar and incubating the plates anaerobically at 37°C for 48 h. In addition, C. perfringens growth from spores was assessed at an isothermal temperature of 44°C. Growth from spores was inhibited in ground beef with a pH of 5.5 or below, even during extended cooling from 54.4 to 7.2°C in 21 h. In ground beef with a pH of 5.6, the growth was >1 log after 18 h of cooling from 54.4 to 7.2°C. However, 15 h of cooling controlled the growth to <1 log, regardless of the inoculum level. In addition, no growth was observed in any product with a pH ranging from 4.74 to 5.17, both during exponential abusive cooling periods of up to 21 h and during storage for 21 h at 44°C. While <1-log growth of C. perfringens from spores was observed in the pH 5.63 product cooled exponentially from 54.4 to 7.2°C in 15 h or less, the pH 6.35 product supported growth, even after 6 h of cooling from 54.4 to 7.2°C. These challenge tests demonstrate that adjustment of ground beef to pH of 5.5 or less and of barbeque products to pH of 5.63 or less inhibits C. perfringens spore germination and outgrowth during extended cooling periods from 54.4 to 7.2°C up to 15 h. Therefore, safe cooling periods for products with homogeneous, lower pHs can be substantially longer. PMID:23317858

  17. 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...

  18. 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

  19. 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

  20. The occurrence of cpb2-toxigenic Clostridium perfringens and the possible role of the ?2-toxin in enteric disease of domestic animals, wild animals and humans

    Microsoft Academic Search

    Georgios N. Nikolaou; Andrea Gröne

    2010-01-01

    The virulence of Clostridium perfringens, a bacterium causing enteritis and enterotoxaemia in domestic and wild animals and humans, results largely from its ability to produce toxins. In 1997, an unknown toxin of C. perfringens, the ?2-toxin, and its encoding gene cpb2 were described. Since that time numerous studies have been published dealing with a possible association of cpb2-harbouring strains of

  1. 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...

  2. Mucin gene mRNA levels in broilers challenged with eimeria and/or Clostridium perfringens.

    PubMed

    Kitessa, Soressa M; Nattrass, Gregory S; Forder, Rebecca E A; McGrice, Hayley A; Wu, Shu-Biao; Hughes, Robert J

    2014-09-01

    The effects of Eimeria (EM) and Clostridium perfringens (CP) challenges on the mRNA levels of genes involved in mucin (Muc) synthesis (Muc2, Muc5ac, Muc13, and trefoil family factor-2 [TFF2]), inflammation (tumor necrosis factor alpha [TNF-alpha] and interleukin-18 [IL-18]), and metabolic processes (cluster of differentiation [CD]36) in the jejunum of broilers were investigated. Two parallel experiments involving 1) EM challenge and 2) EM and CP challenges were conducted. The first experiment was a 2 X 2 study with 12 birds per treatment (N = 48) involving fishmeal substitution (25%) in the diet (FM) and EM challenge. The treatments were: Control (FM-, EM-), Fishmeal (FM+, EM-), EM challenge (FM-, EM+), and fishmeal substitution and EM challenge (FM+, EM+). The second experiment was a 2 X 2 X 2 experiment with six birds per treatment (N = 48) involving fishmeal (FM-, FM+), Eimeria (EM-, EM+), and C perfringens (CP-, CP+). In both arms of the study, male broilers were given a starter diet for the whole period of 16 days, except those assigned to FM+, where 25% of the starter ration was replaced with fishmeal from days 8 to 14. EM inoculation was performed on day 9 and CP inoculation on days 14 and 15. The EM challenge birds were euthanatized for sampling on day 13; postmortem examination and sampling for the Eimeria plus C perfringens challenge arm of the study were on day 16. In the Eimeria challenge arm of the study, fishmeal supplementation significantly suppressed the mRNA levels of TNF-alpha, TFF2, and IL-18 pre-CP inoculation but simultaneously increased the levels of Muc13 and CD36 mRNAs. Birds challenged with Eimeria exhibited increased mRNA levels of Muc13, Muc5ac, TNF-alpha, and IL-18. In the Eimeria and C. perfringens challenge arm, birds exposed to EM challenge exhibited significantly lower mRNA levels of Muc2 and CD36. The mRNA levels of CD36 were also significantly suppressed by CP challenge. Our results showed that the transcription of mucin synthesis genes in the jejunum of broilers is modulated by fishmeal inclusion in the diet. Furthermore, we show for the first time suppression of CD36 mRNA levels in the intestine of broilers challenged with Eimeria or C. perfringens. PMID:25518436

  3. Isolation of a plasmid responsible for caseinase activity in Clostridium perfringens ATCC 3626B.

    PubMed Central

    Blaschek, H P; Solberg, M

    1981-01-01

    Clostridium perfringens strain ATCC 3626B was cured of caseinase activity at a high frequency after treatment with acriflavine dye (2.5%) or elevated temperature growth (9.1%). Caseinase-negative isolates retained the larger (9.4 megadaltons) pHB102 cryptic plasmid, but were missing the smaller (2.1 megadaltons) pHB101 plasmid present in the caseinase-positive wild-type strain. Dye-buoyant density-gradient centrifugation at 4 or 15 degrees C revealed that the pHB101 and pHB102 plasmids are temperature labile and easily converted into the nicked non-supercoiled or linear state. Images PMID:6263868

  4. Tight junctions. Structural insight into tight junction disassembly by Clostridium perfringens enterotoxin.

    PubMed

    Saitoh, Yasunori; Suzuki, Hiroshi; Tani, Kazutoshi; Nishikawa, Kouki; Irie, Katsumasa; Ogura, Yuki; Tamura, Atsushi; Tsukita, Sachiko; Fujiyoshi, Yoshinori

    2015-02-13

    The C-terminal region of Clostridium perfringens enterotoxin (C-CPE) can bind to specific claudins, resulting in the disintegration of tight junctions (TJs) and an increase in the paracellular permeability across epithelial cell sheets. Here we present the structure of mammalian claudin-19 in complex with C-CPE at 3.7 Å resolution. The structure shows that C-CPE forms extensive hydrophobic and hydrophilic interactions with the two extracellular segments of claudin-19. The claudin-19/C-CPE complex shows no density of a short extracellular helix that is critical for claudins to assemble into TJ strands. The helix displacement may thus underlie C-CPE-mediated disassembly of TJs. PMID:25678664

  5. Purification and characterization of an extracellular alpha-amylase from Clostridium perfringens type A.

    PubMed Central

    Shih, N J; Labbé, R G

    1995-01-01

    An alpha-amylase (EC 3.2.1.1) secreted by Clostridium perfringens NCTC 8679 type A was purified to homogeneity and characterized. It was isolated from concentrated cell-free culture medium by ion-exchange and gel permeation chromatography. The enzyme exhibited maximal activity at pH 6.5 and 30 degrees C without the presence of calcium. The pI of the enzyme was 4.75. The estimated molecular weight of the purified enzyme was 76 kDa. The purified enzyme was inactivated between 35 and 40 degrees C, which increased to between 45 and 50 degrees C in the presence of calcium (5 mM). The purified enzyme produced a mixture of oligosaccharides as major end products of starch hydrolysis, indicating alpha-amylase activity. PMID:7646015

  6. 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

  7. Vaccination with recombinant Clostridium perfringens toxoids ? and ? promotes elevated antepartum and passive humoral immunity in swine.

    PubMed

    Salvarani, Felipe M; Conceição, Fabricio R; Cunha, Carlos E P; Moreira, Gustavo M S G; Pires, Prhiscylla S; Silva, Rodrigo O S; Alves, Guilherme G; Lobato, Francisco C F

    2013-08-28

    Due to the increasingly restricted use of antimicrobials in animal production systems, the prevention and control of Clostridium perfringens type A- and C-induced diarrhea in piglets should be based on passive immunization via the prepartum vaccination of sows. Given the current obstacles in the production of conventional clostridial vaccines, the use of recombinant proteins has been considered to represent a promising alternative. In the present study, the neutralizing antibody response of immunized sows and their litters to a bivalent vaccine containing the C. perfringens recombinant toxoids alpha (rTA) and beta (rTB) produced in Escherichia coli was assessed. Rabbits (n=8) and pregnant sows (n=7) were immunized with 200?g of each recombinant antigen using Al(OH)3 as adjuvant. The alpha and beta antitoxin titer detected in the rabbits' serum pool was 9.6 and 20.4IU/mL, respectively. The mean alpha and beta antitoxin titers in the sows' sera were 6.0±0.9IU/mL and 14.5±2.2IU/mL, and the corresponding individual coefficients of variation (CV) were 16.04% and 14.91%, respectively. The mean alpha and beta antitoxin titers in the litters' serum pools were 4.2±0.4IU/mL and 10.9±1.7IU/mL, and the CV between litters was 9.23% and 9.85%, respectively. The results showed that the rTA and rTB proteins produced and tested in the present study induced an immune response and can be regarded as candidates for the development of a commercial vaccine against C. perfringens type A- and C-induced diarrhea in pigs. PMID:23845812

  8. How do different concentrations of Clostridium perfringens affect the quality of extended boar spermatozoa?

    PubMed

    Sepúlveda, Lilian; Bussalleu, Eva; Yeste, Marc; Torner, Eva; Bonet, Sergi

    2013-07-01

    Bacteriospermia in boar fresh and extended semen is a frequent finding that produces alterations on sperm quality and, consequently, causes economic losses in artificial insemination (AI) centres. The present study sought to evaluate the effect of different infective concentrations of Clostridium perfringens on boar sperm quality, assessed as sperm motility (CASA), morphology and viability, through 11 days of storage at 15°C (experiment 1), and after 96h of incubation at 37°C (experiment 2). With this purpose, different seminal doses were artificially inoculated with different infective concentrations of C. perfringens, ranging from 10(2) to 10(8)cfumL(-1). The negative controls were non-inoculated doses. Sperm quality was checked after 0, 1, 2, 3, 4, 7, 8, 9, 10 and 11 days of storage at 15°C in experiment 1, and after 0, 24, 48, 72 and 96h at 37°C in the second experiment. Moreover, the presence/absence of bacteria was detected by PCR analyses during both experiments at different time points. In both experiments, sperm morphology of inoculated samples did not differ from the negative control. Conversely, detrimental effects on sperm viability and motility were observed after 24h of incubation/storage at the highest infective concentrations in both experiments. The deleterious effects observed because of the presence of C. perfringens in semen emphasise the relevance of detecting bacteria in extended doses destined to AI. So, this study suggests that the evaluation of bacterial contamination in semen is a procedure that should be routinely applied while assessing sperm quality in AI centres to avoid the use of doses with low sperm quality and the possible spread of bacterial contaminants. PMID:23755936

  9. 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

  10. 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

  11. Effect of fluoroquinolone resistance selection on the fitness of three strains of Clostridium perfringens.

    PubMed

    Park, Miseon; Sutherland, John B; Kim, Jong Nam; Rafii, Fatemeh

    2013-12-01

    Selection of bacterial strains for resistance to antimicrobial agents may affect fitness; and fluoroquinolone resistance has been shown to affect fitness of aerobic and facultative bacteria. The impact on bacterial fitness of resistance selection to three fluoroquinolones was examined in three wild-type strains of Clostridium perfringens: ATCC 13124, ATCC 3626, and NCTR. Selection for resistance to norfloxacin, ciprofloxacin, and gatifloxacin affected the fitness of nine mutant strains differently. In a series of pure cultures grown in the absence of drugs, the growth of each of the mutants was comparable to that of the corresponding wild type. In competition experiments between mutants and isogenic wild types, however, some of the mutants were less fit. The fitness of ciprofloxacin-resistant mutants was comparable for all three strains, but two gatifloxacin-resistant mutants and one norfloxacin-resistant mutant were significantly less fit than the corresponding wild types. We conclude that both the genetic background of the strain and the fluoroquinolone which induced resistance affected the fitness of the resistant mutants. This is the first time that the effect on fitness of resistance to antimicrobial agents has been measured for C. perfringens. PMID:23789809

  12. Biochemistry and Physiology of the ? Class Carbonic Anhydrase (Cpb) from Clostridium perfringens Strain 13

    PubMed Central

    Kumar, R. Siva Sai; Hendrick, William; Correll, Jared B.; Patterson, Andrew D.; Melville, Stephen B.

    2013-01-01

    The carbonic anhydrase (Cpb) from Clostridium perfringens strain 13, the only carbonic anhydrase encoded in the genome, was characterized both biochemically and physiologically. Heterologously produced and purified Cpb was shown to belong to the type I subclass of the ? class, the first ? class enzyme investigated from a strictly anaerobic species of the domain Bacteria. Kinetic analyses revealed a two-step, ping-pong, zinc-hydroxide mechanism of catalysis with Km and kcat/Km values of 3.1 mM CO2 and 4.8 × 106 s?1 M?1, respectively. Analyses of a cpb deletion mutant of C. perfringens strain HN13 showed that Cpb is strictly required for growth when cultured in semidefined medium and an atmosphere without CO2. The growth of the mutant was the same as that of the parent wild-type strain when cultured in nutrient-rich media with or without CO2 in the atmosphere, although elimination of glucose resulted in decreased production of acetate, propionate, and butyrate. The results suggest a role for Cpb in anaplerotic CO2 fixation reactions by supplying bicarbonate to carboxylases. Potential roles in competitive fitness are discussed. PMID:23475974

  13. Comparative Neuropathology of Ovine Enterotoxemia Produced by Clostridium perfringens Type D Wild-Type Strain CN1020 and Its Genetically Modified Derivatives.

    PubMed

    Garcia, J P; Giannitti, F; Finnie, J W; Manavis, J; Beingesser, J; Adams, V; Rood, J I; Uzal, F A

    2014-06-25

    Clostridium perfringens type D causes enterotoxemia in sheep and goats. The disease is mediated by epsilon toxin (ETX), which affects the cerebrovascular endothelium, increasing vascular permeability and leading to cerebral edema. In the present study, we compared the distribution and severity of the cerebrovascular changes induced in lambs by C. perfringens type D strain CN1020, its isogenic etx null mutant, and the ETX-producing complemented mutant. We also applied histochemical and immunohistochemical markers to further characterize the brain lesions induced by ETX. Both ETX-producing strains induced extensive cerebrovascular damage that did not differ significantly between each other in nature, neuroanatomic distribution, or severity. By contrast, lambs inoculated with the etx mutant or sterile, nontoxic culture medium did not develop detectable brain lesions, confirming that the neuropathologic effects observed in these infections are dependent on ETX production. Lambs treated with the wild-type and complemented strains showed perivascular and mural vascular edema, as well as serum albumin extravasation, particularly severe in the cerebral white matter, midbrain, medulla oblongata, and cerebellum. Brains of animals inoculated with the ETX-producing strains showed decreased expression of glial fibrillary acidic protein and increased expression of aquaporin-4 in the end-feet processes of the astrocytes around blood vessels. Early axonal injury was demonstrated with anti-amyloid precursor protein immunohistochemistry. Perivascular accumulation of macrophages/microglia with intracytoplasmic albumin globules was also observed in these animals. This study demonstrates that ETX is responsible for the major cerebrovascular changes in C. perfringens type D-induced disease. PMID:24964921

  14. 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

  15. Clostridium perfringens enterotoxin carboxy-terminal fragment is a novel tumor-homing peptide for human ovarian cancer

    Microsoft Academic Search

    Emiliano Cocco; Francesca Casagrande; Stefania Bellone; Christine E Richter; Marta Bellone; Paola Todeschini; Jennie C Holmberg; Han Hsuan Fu; Michele K Montagna; Gil Mor; Peter E Schwartz; Dan Arin-Silasi; Masoud Azoudi; Thomas J Rutherford; Maysa Abu-Khalaf; Sergio Pecorelli; Alessandro D Santin

    2010-01-01

    BACKGROUND: Development of innovative, effective therapies against recurrent\\/chemotherapy-resistant ovarian cancer remains a high priority. Using high-throughput technologies to analyze genetic fingerprints of ovarian cancer, we have discovered extremely high expression of the genes encoding the proteins claudin-3 and claudin-4. METHODS: Because claudin-3 and -4 are the epithelial receptors for Clostridium perfringens enterotoxin (CPE), and are sufficient to mediate CPE binding,

  16. 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

  17. Proteome and Transcriptome Analysis of the Virulence Genes Regulated by the VirR\\/VirS System in Clostridium perfringens

    Microsoft Academic Search

    Takeshi Shimizu; Kensuke Shima; Ken-ichi Yoshino; Kazuyoshi Yonezawa; Tohru Shimizu; Hideo Hayashi

    2002-01-01

    The proteins under the control of the two-component system VirR\\/VirS in Clostridium perfringens were analyzed by using two-dimensional gel electrophoresis of the culture supernatant from the wild type and the virR mutant. Based on matrix-assisted laser desorption ionization-time of flight\\/mass spectrometry, seven positively regulated proteins and eight negatively regulated proteins were identified. Transcriptome analysis confirmed that 7 of the 15

  18. Effects of Clostridium perfringens enterotoxin via claudin-4 on normal human pancreatic duct epithelial cells and cancer cells

    Microsoft Academic Search

    Hiroshi Yamaguchi; Takashi Kojima; Tatsuya Ito; Daisuke Kyuno; Yasutoshi Kimura; Masafumi Imamura; Koichi Hirata; Norimasa Sawada

    The tight junction protein claudin-4 is frequently overexpressed in pancreatic cancer, and is also a receptor for Clostridium perfringens enterotoxin (CPE). The cytotoxic effects of CPE are thought to be useful as a novel therapeutic tool for pancreatic cancer.\\u000a However, the responses to CPE via claudin-4 remain unknown in normal human pancreatic duct epithelial (HPDE) cells. We introduced\\u000a the human

  19. Determination of the effect of single abomasal or jejunal inoculation of Clostridium perfringens Type A in dairy cows

    PubMed Central

    2005-01-01

    Abstract A randomized study was conducted to determine if inoculation of the abomasum or jejunum with Clostridium perfringens Type A would induce jejunal hemorrhage syndrome in healthy cows. Twelve adult nonlactating dairy cows were inoculated with 10 mL of pure culture broth of C. perfringens type A (beta2 toxin positive) into the abomasum (n = 6) or jejunum (n = 6). On day 6, the cows were euthanized and samples for culture were taken from the abomasum, jejunum, and feces. No cows developed clinical signs of jejunal hemorrhage syndrome during the course of the study. Five of 6 abomasal samples and 1 of 6 jejunal samples were positive for C. perfringens Type A (beta2 negative) prior to inoculation. Eight of 12 abomasal samples, 11 of 12 fecal samples, and 10 of 12 jejunal samples were positive for C. perfringens Type A (beta2 negative) after inoculation. Intraluminal inoculation of C. perfringens Type A alone at this dose and under these conditions did not induce clinical signs of jejunal hemorrhage syndrome in adult dairy cows. The multifactorial nature of the disease likely contributed to our inability to reproduce the disease in this study. PMID:16231652

  20. Differential Responses of Cecal Microbiota to Fishmeal, Eimeria and Clostridium perfringens in a Necrotic Enteritis Challenge Model in Chickens

    PubMed Central

    Rodgers, Nicholas; Swick, Robert A.; Moore, Robert J.

    2014-01-01

    Clostridium perfringens causes enteric diseases in animals and humans. In poultry, avian-specific C. perfringens strains cause necrotic enteritis, an economically significant poultry disease that costs the global industry over $2 billion annually in losses and control measures. With removal of antibiotic growth promoters in some countries this disease appears to be on the rise. In experimental conditions used to study disease pathogenesis and potential control measures, reproduction of the disease relies on the use of predisposing factors such as Eimeria infection and the use of high protein diets, indicating complex mechanisms involved in the onset of necrotic enteritis. The mechanisms by which the predisposing factors contribute to disease progression are not well understood but it has been suggested that they may cause perturbations in the microbiota within the gastrointestinal tract. We inspected changes in cecal microbiota and short chain fatty acids (SCFA) induced by Eimeria and fishmeal, in birds challenged or not challenged with C. perfringens. C. perfringens challenge in the absence of predisposing factors did not cause significant changes in either the alpha or beta diversity of the microbiota nor in concentrations of SCFA. Moreover, there was no C. perfringens detected in the cecal microbiota 2 days post-challenge without the presence of predisposing factors. In contrast, both fishmeal and Eimeria caused significant changes in microbiota, seen in both alpha and beta diversity and also enabled C. perfringens to establish itself post challenge. Eimeria had its strongest influence on intestinal microbiota and SCFA when combined with fishmeal. Out of 6 SCFAs measured, including butyric acid, none were significantly influenced by C. perfringens, but their levels were strongly modified following the use of both predisposing factors. There was little overlap in the changes caused following Eimeria and fishmeal treatments, possibly indicating multiple routes for progressing towards clinical symptoms of necrotic enteritis. PMID:25167074

  1. Perfrin, a novel bacteriocin associated with netB positive Clostridium perfringens strains from broilers with necrotic enteritis.

    PubMed

    Timbermont, Leen; De Smet, Lina; Van Nieuwerburgh, Filip; Parreira, Valeria R; Van Driessche, Gonzalez; Haesebrouck, Freddy; Ducatelle, Richard; Prescott, John; Deforce, Dieter; Devreese, Bart; Van Immerseel, Filip

    2014-01-01

    Necrotic enteritis in broiler chickens is associated with netB positive Clostridium perfringens type A strains. It is known that C. perfringens strains isolated from outbreaks of necrotic enteritis are more capable of secreting factors inhibiting growth of other C. perfringens strains than strains isolated from the gut of healthy chickens. This characteristic could lead to extensive and selective presence of a strain that contains the genetic make-up enabling to secrete toxins that cause gut lesions. This report describes the discovery, purification, characterization and recombinant expression of a novel bacteriocin, referred to as perfrin, produced by a necrotic enteritis-associated netB-positive C. perfringens strain. Perfrin is a 11.5 kDa C-terminal fragment of a 22.9 kDa protein and showed no sequence homology to any currently known bacteriocin. The 11.5 kDa fragment can be cloned into Escherichia coli, and expression yielded an active peptide. PCR detection of the gene showed its presence in 10 netB-positive C. perfringens strains of broiler origin, and not in other C. perfringens strains tested (isolated from broilers, cattle, sheep, pigs, and humans). Perfrin and NetB are not located on the same genetic element since NetB is plasmid-encoded and perfrin is not. The bacteriocin has bactericidal activity over a wide pH-range but is thermolabile and sensitive to proteolytic digestion (trypsin, proteinase K). C. perfringens bacteriocins, such as perfrin, can be considered as an additional factor involved in the pathogenesis of necrotic enteritis in broilers. PMID:24708344

  2. Comparative transcription analysis and toxin production of two fluoroquinolone-resistant mutants of Clostridium perfringens

    PubMed Central

    2013-01-01

    Background Fluoroquinolone use has been listed as a risk factor for the emergence of virulent clinical strains of some bacteria. The aim of our study was to evaluate the effect of fluoroquinolone (gatifloxacin) resistance selection on differential gene expression, including the toxin genes involved in virulence, in two fluoroquinolone-resistant strains of Clostridium perfringens by comparison with their wild-type isogenic strains. Results DNA microarray analyses were used to compare the gene transcription of two wild types, NCTR and ATCC 13124, with their gatifloxacin-resistant mutants, NCTRR and 13124R. Transcription of a variety of genes involved in bacterial metabolism was either higher or lower in the mutants than in the wild types. Some genes, including genes for toxins and regulatory genes, were upregulated in NCTRR and downregulated in 13124R. Transcription analysis by quantitative real-time PCR (qRT-PCR) confirmed the altered expression of many of the genes that were affected differently in the fluoroquinolone-resistant mutants and wild types. The levels of gene expression and enzyme production for the toxins phospholipase C, perfringolysin O, collagenase and clostripain had decreased in 13124R and increased in NCTRR in comparison with the wild types. After centrifugation, the cytotoxicity of the supernatants of NCTRR and 13224R cultures for mouse peritoneal macrophages confirmed the increased cytotoxicity of NCTRR and the decreased cytotoxicity of 13124R in comparison with the respective wild types. Fluoroquinolone resistance selection also affected cell shape and colony morphology in both strains. Conclusion Our results indicate that gatifloxacin resistance selection was associated with altered gene expression in two C. perfringens strains and that the effect was strain-specific. This study clearly demonstrates that bacterial exposure to fluoroquinolones may affect virulence (toxin production) in addition to drug resistance. PMID:23452396

  3. Structural and Functional Analysis of the Pore-Forming Toxin NetB from Clostridium perfringens

    PubMed Central

    Yan, Xu-Xia; Porter, Corrine J.; Hardy, Simon P.; Steer, David; Smith, A. Ian; Quinsey, Noelene S.; Hughes, Victoria; Cheung, Jackie K.; Keyburn, Anthony L.; Kaldhusdal, Magne; Moore, Robert J.; Bannam, Trudi L.; Whisstock, James C.; Rood, Julian I.

    2013-01-01

    ABSTRACT Clostridium perfringens is an anaerobic bacterium that causes numerous important human and animal diseases, primarily as a result of its ability to produce many different protein toxins. In chickens, C. perfringens causes necrotic enteritis, a disease of economic importance to the worldwide poultry industry. The secreted pore-forming toxin NetB is a key virulence factor in the pathogenesis of avian necrotic enteritis and is similar to alpha-hemolysin, a ?-barrel pore-forming toxin from Staphylococcus aureus. To address the molecular mechanisms underlying NetB-mediated tissue damage, we determined the crystal structure of the monomeric form of NetB to 1.8 Å. Structural comparisons with other members of the alpha-hemolysin family revealed significant differences in the conformation of the membrane binding domain. These data suggested that NetB may recognize different membrane receptors or use a different mechanism for membrane-protein interactions. Consistent with this idea, electrophysiological experiments with planar lipid bilayers revealed that NetB formed pores with much larger single-channel conductance than alpha-hemolysin. Channel conductance varied with phospholipid net charge. Furthermore, NetB differed in its ion selectivity, preferring cations over anions. Using hemolysis as a screen, we carried out a random-mutagenesis study that identified several residues that are critical for NetB-induced cell lysis. Mapping of these residues onto the crystal structure revealed that they were clustered in regions predicted to be required for oligomerization or membrane binding. Together these data provide an insight into the mechanism of NetB-mediated pore formation and will contribute to our understanding of the mode of action of this important toxin. PMID:23386432

  4. Genotyping and Phenotyping of Beta2-Toxigenic Clostridium perfringens Fecal Isolates Associated with Gastrointestinal Diseases in Piglets

    PubMed Central

    Waters, Michael; Savoie, Amanda; Garmory, Helen S.; Bueschel, Dawn; Popoff, Michel R.; Glenn Songer, J.; Titball, Richard W.; McClane, Bruce A.; Sarker, Mahfuzur R.

    2003-01-01

    Although Clostridium perfringens is recognized as an important cause of clostridial enteric diseases, only limited knowledge exists concerning the association of particular C. perfringens toxinotypes (type A to E) with gastrointestinal (GI) diseases in domestic animals. Some C. perfringens isolates also produce the newly discovered beta2-toxin (CPB2). Recent epidemiological studies suggested that C. perfringens isolates carrying the gene encoding CPB2 (cpb2) are strongly associated with clostridial GI diseases in domestic animals, including necrotic enteritis in piglets and typhlocolitis in horses. These putative relationships, obtained by PCR genotyping, were tested in the present study by further genotyping and phenotyping of 29 cpb2-positive C. perfringens isolates from pigs with GI disease (pig GI disease isolates). PCR and restriction fragment length polymorphism analysis reconfirmed the presence of cpb2 gene sequences in all the disease isolates included in the study. Furthermore, genotyping by pulsed-field gel electrophoresis analyses showed that the pig GI disease isolates included in this study all carry a plasmid cpb2 gene, yet no clonal relationships were detected between the cpb2-positive pig GI disease isolates surveyed. Finally, CPB2-specific Western blotting demonstrated CPB2 expression by all of the cpb2-positive isolates surveyed. The CPB2 proteins made by five of these pig GI disease isolates were shown to have the same deduced amino acid sequences as the biologically active CPB2 protein made by the original type C isolate, CWC245. Collectively, our present results support a significant association between CPB2-positive C. perfringens isolates and diarrhea in piglets. PMID:12904359

  5. Inflammatory responses to a Clostridium perfringens type A strain and ?-toxin in primary intestinal epithelial cells of chicken embryos.

    PubMed

    Guo, Shuangshuang; Li, Changwu; Liu, Dan; Guo, Yuming

    2015-04-01

    The causative pathogen of necrotic enteritis is the Gram-positive bacterium Clostridium perfringens. Its main cell wall component, peptidoglycan (PGN), can be recognized by Toll-like receptor 2 and nucleotide-binding oligomerization domain (NOD). Consequently, the immune response is initiated via activation of nuclear factor kappa B (NF-?B) signalling pathway. An in vitro study was conducted to investigate chicken intestinal inflammatory responses to C. perfringens type A and one of its virulence factors, ?-toxin. In primary intestinal epithelial cells, C. perfringens as well as commercially available PGN and ?-toxin challenge upregulated mRNA expression of interleukin (IL)-6, IL-8 and inducible nitric oxide synthase (iNOS) with a dosage-dependent manner at 3 h post infection (p.i.; P ? 0.001). Time-course effects of three stimulators at high concentration were further examined. C. perfringens infection elevated IL-6, IL-8 and iNOS levels from 1 h to 9 h p.i., while PGN treatment increased IL-6 and IL-8 expression at 1 h and 3 h p.i. (P < 0.05). Bacterial and PGN treatments induced NOD1 expression at 6 h p.i. and only bacterial infection boosted NF-?B p65 expression at 6 h and 9 h p.i. (P < 0.05). ?-Toxin treatment upregulated IL-6 and IL-8 expression throughout infection, as well as iNOS, TNF-? and NF-?B p65 expression at later hours p.i. (P < 0.05). In conclusion, both C. perfringens and ?-toxin challenge induced intense cytokine expression associated with NF-?B activation in chicken intestinal epithelial cells. The receptors for the recognition of PGN component of C. perfringens need further investigation. PMID:25584964

  6. Clostridium perfringens Iota Toxin: Binding Studies and Characterization of Cell Surface Receptor by Fluorescence-Activated Cytometry

    PubMed Central

    Stiles, Bradley G.; Hale, Martha L.; Marvaud, Jean-Christophe; Popoff, Michel R.

    2000-01-01

    The binding characteristics of iota toxin, a binary enterotoxin produced by Clostridium perfringens type E, were studied by fluorescence-activated cytometry. The proteolytically activated binding component of iota toxin, iota b (Ib), bound to various cell types when incubated at 4, 25, or 37°C for 10 min. The binding of Ib was inhibited by antisera against C. perfringens type E or Clostridium spiroforme culture supernatants, but not C. perfringens types C or D. Pretreatment of Vero cells with glycosidases or lectins did not affect Ib interactions, while pronase effectively prevented Ib binding to the cell surface. The Ib protomer (Ibp) bound to the cell surface, but trypsinization of Ibp was necessary for docking of the ADP-ribosylating component, iota a (Ia). Ia attached to cell-bound Ib within 10 min at 37°C, but surface levels of Ia decreased 90% after 30 min and were undetectable by 60 min. Detectable surface levels of Ib also diminished over time, and Western blot analysis suggested internalization or embedment of Ib into the membrane. PMID:10816501

  7. Molecular variation between the  -toxins from the type strain (NCTC 8237) and clinical isolates of Clostridium perfringens associated with disease in man and animals

    Microsoft Academic Search

    A. Ginter; E. D. Williamson; F. Dessy; P. Coppe; H. Bullifent; A. Howells; R. W. Titball

    1996-01-01

    The a-toxin produced by the type strain of Clostridium perfringens (NCTC 8237) was shown to differ from the a-toxins produced by most strains of C. perfringens isolated from man and from calves with respect to reactivity with a neutralizing monoclonal antibody (DY2F5D11). The difference in antibody binding correlated with three differences in the deduced amino acid sequence (Ala174 to Asp,,;

  8. Multi-Organ Failure Secondary to a Clostridium Perfringens Gaseous Liver Abscess following a Self-Limited Episode of Acute Gastroenteritis.

    PubMed

    Eltawansy, Sherif Ali; Merchant, Chandni; Atluri, Paavani; Dwivedi, Sukrut

    2015-01-01

    Background Clostridium perfringens is an unusual pathogen responsible for the development of a gas-forming pyogenic liver abscess. Progression to septicemia with this infection has amplified case fatality rates. Case Report We report a case of an 81-year-old lady with pyogenic liver abscess with gas formation that was preceded by an acute gastroenteritis. The most common precipitating factors are invasive procedures and immunosuppression. Clostridium perfringens was unexpectedly isolated in the drained abscess, as well as blood. It is a normal inhabitant of the human bowel and a common cause of food poisoning, notoriously leading to tissue necrosis and gas gangrene. Conclusions We report a case of gas-forming pyogenic liver abscess and bacteremia progressing to fatal septic shock, caused by an uncommon Clostridium perfringens isolate. PMID:25807198

  9. Multi-Organ Failure Secondary to a Clostridium Perfringens Gaseous Liver Abscess following a Self-Limited Episode of Acute Gastroenteritis

    PubMed Central

    Eltawansy, Sherif Ali; Merchant, Chandni; Atluri, Paavani; Dwivedi, Sukrut

    2015-01-01

    Patient: Female, 81 Final Diagnosis: Liver absces Symptoms: Diarrhea • jaundice • vomiting • weakness Medication: — Clinical Procedure: CT scan guided drainage Specialty: Gastroenterology and Hepatology Objective: Rare disease Background: Clostridium perfringens is an unusual pathogen responsible for the development of a gas-forming pyogenic liver abscess. Progression to septicemia with this infection has amplified case fatality rates. Case Report: We report a case of an 81-year-old lady with pyogenic liver abscess with gas formation that was preceded by an acute gastroenteritis. The most common precipitating factors are invasive procedures and immunosuppression. Clostridium perfringens was unexpectedly isolated in the drained abscess, as well as blood. It is a normal inhabitant of the human bowel and a common cause of food poisoning, notoriously leading to tissue necrosis and gas gangrene. Conclusions: We report a case of gas-forming pyogenic liver abscess and bacteremia progressing to fatal septic shock, caused by an uncommon Clostridium perfringens isolate. PMID:25807198

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

    PubMed

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

    2014-04-01

    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 the introduction of C. perfringens. Our results indicate that EcN can inhibit many of the pathological effects of C. perfringens in vitro conditions. PMID:24532573

  11. 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

  12. Internalization of Clostridium perfringens ?-toxin leads to ERK activation and is involved on its cytotoxic effect.

    PubMed

    Monturiol-Gross, Laura; Flores-Díaz, Marietta; Campos-Rodríguez, Diana; Mora, Rodrigo; Rodríguez-Vega, Mariela; Marks, David L; Alape-Girón, Alberto

    2014-04-01

    Clostridium perfringens phospholipase C (CpPLC), also called ?-toxin, plays a key role in the pathogenesis of gas gangrene. CpPLC may lead to cell lysis at concentrations that cause extensive degradation of plasma membrane phospholipids. However, at sublytic concentrations it induces cytotoxicity without inducing evident membrane damage. The results of this work demonstrate that CpPLC becomes internalized in cells by a dynamin-dependent mechanism and in a time progressive process: first, CpPLC colocalizes with caveolin both at the plasma membrane and in vesicles, and later it colocalizes with early and late endosomes and lysosomes. Lysosomal damage in the target cells is evident 9?h after CpPLC exposure. Our previous work demonstrated that CpPLCinduces ERK1/2 activation, which is involved in its cytotoxic effect. In this work we found that cholesterol sequestration, dynamin inhibition, as well as inhibition of actin polymerization, prevent CpPLC internalization and ERK1/2 activation, involving endocytosis in the signalling events required for CpPLC cytotoxic effect at sublytic concentrations. These results provide new insights about the mode of action of this bacterial phospholipase C, previously considered to act only locally on cell membrane. PMID:24245664

  13. Molecular cloning and nucleotide sequence of the alpha-toxin (phospholipase C) of Clostridium perfringens.

    PubMed Central

    Titball, R W; Hunter, S E; Martin, K L; Morris, B C; Shuttleworth, A D; Rubidge, T; Anderson, D W; Kelly, D C

    1989-01-01

    A fragment of DNA containing the gene coding for the phospholipase C (alpha-toxin) of Clostridium perfringens was cloned into Escherichia coli. The cloned DNA appeared to code only for the alpha-toxin and contained both the coding region and its associated gene promoter. The nucleotide sequence of the cloned DNA was determined, and an open reading frame was identified which encoded a protein with a molecular weight of 42,528. By comparison of the gene sequence with the N-terminal amino acid sequence of the protein, a 28-amino-acid signal sequence was identified. The gene promoter showed considerable homology with the E. coli sigma 55 consensus promoter sequences, and this may explain why the gene was expressed by E. coli. The cloned gene product appeared to be virtually identical to the native protein. A 77-amino-acid stretch that was close to the N terminus of the alpha-toxin showed considerable homology with similarly located regions of the Bacillus cereus phosphatidylcholine, preferring phospholipase C and weaker homology with the phospholipase C from Pseudomonas aeruginosa. Images PMID:2536355

  14. Death Pathways Activated in CaCo-2 Cells by Clostridium perfringens Enterotoxin

    PubMed Central

    Chakrabarti, Ganes; Zhou, Xin; McClane, Bruce A.

    2003-01-01

    Clostridium perfringens enterotoxin (CPE), a 35-kDa polypeptide, induces cytotoxic effects in the enterocyte-like CaCo-2 cell culture model. To identify the mammalian cell death pathway(s) mediating CPE-induced cell death, CaCo-2 cultures were treated with either 1 or 10 ?g of CPE per ml. Both CPE doses were found to induce morphological damage and DNA cleavage in CaCo-2 cells. The oncosis inhibitor glycine, but not a broad-spectrum caspase inhibitor, was able to transiently block both of those pathological effects in CaCo-2 cells treated with the higher, but not the lower, CPE dose. Conversely, a caspase 3/7 inhibitor (but not glycine or a caspase 1 inhibitor) blocked morphological damage and DNA cleavage in CaCo-2 cells treated with the lower, but not the higher, CPE dose. Collectively, these results indicate that lower CPE doses cause caspase 3/7-dependent apoptosis, while higher CPE doses induce oncosis. Apoptosis caused by the lower CPE dose was shown to proceed via a classical pathway involving mitochondrial membrane depolarization and cytochrome c release. As the CPE concentrations used in this study for demonstrating apoptosis and oncosis have pathophysiologic relevance, these results suggest that both oncosis and apoptosis may occur in the intestines during CPE-associated gastrointestinal disease. PMID:12874301

  15. The role of penicillin G potassium in managing Clostridium perfringens in broiler chickens.

    PubMed

    Gadbois, P; Brennan, J J; Bruce, L; Wilson, J B; Aramini, J J

    2008-09-01

    The efficacy of penicillin G potassium (Pot-Pen) administered via drinking water to manage necrotic enteritis (NE) was investigated in a Clostridium perfringens (CP) challenge study using 1600 broiler chickens assigned to one of four treatment groups: nonchallenged, nonmedicated; challenged, nonmedicated; challenged, Pot-Pen 0.2 g/L; challenged, Pot-Pen 0.4 g/L. Overall mortality due to NE was significantly reduced among Pot-Pen-treated pens; mortality due to other causes did not differ among the treatment groups. Among all birds, growth performance parameters were significantly improved among Pot-Pen-treated pens. When considering birds randomly sacrificed 4 days post-Pot-Pen initiation, mean NE lesion scores were greatest among the challenged, nonmedicated pens; only one of 80 randomly sacrificed birds treated with Pot-Pen had NE lesions. Among the nonmedicated control pens, body weight (BW) was significantly greater among birds that did not have NE-associated lesions. When sacrificed birds were stratified by NE lesion score, there were no significant differences in BW among the treatment groups. Results of this study suggest that CP-associated subclinical disease can significantly reduce broiler performance. Furthermore, the positive effects of treatment with Pot-Pen appeared to be associated with the prevention and/or treatment of NE-specific lesions. PMID:18939627

  16. Recent progress in understanding the pathogenesis of Clostridium perfringens type C infections

    PubMed Central

    McClane, B. A.

    2011-01-01

    Clostridium perfringens type C causes necrotizing enteritis in humans and several other animal species. Type C isolates must produce at least beta toxin (CPB) and alpha toxin (CPA) and most strains produce several other toxins including perfringolysin O (PFO) and TpeL. However, current evidence indicates that CPB is the main virulence factor for type C infections. Most of this evidence is based upon the loss of virulence shown by isogenic type C CPB knock out mutants on cells, and also in rabbit intestinal loops and in mouse models. This virulence is regained when these mutants are complemented with the wild-type cpb gene. Many type C isolates respond to close contact with enterocyte-like Caco-2 cells by producing all toxins, except TpeL, much more rapidly than occurs during in vitro growth. This in vivo effect involves rapid transcriptional upregulation of the cpb, cpb2, pfoA and plc toxin genes. Rapid Caco-2 cell-induced upregulation of CPB and PFO production involves the VirS/VirR two-component system, since upregulated in vivo transcription of the pfoA and cpb genes was blocked by inactivating the virR gene and was reversible by complementation to restore VirR expression. PMID:21420802

  17. The p38 MAPK and JNK Pathways Protect Host Cells against Clostridium perfringens Beta-Toxin

    PubMed Central

    Shibutani, Masahiro; Seike, Soshi; Yonezaki, Mami; Takagishi, Teruhisa; Oda, Masataka; Kobayashi, Keiko; Sakurai, Jun

    2013-01-01

    Clostridium perfringens beta-toxin is an important agent of necrotic enteritis and enterotoxemia. Beta-toxin is a pore-forming toxin (PFT) that causes cytotoxicity. Two mitogen-activated protein kinase (MAPK) pathways (p38 and c-Jun N-terminal kinase [JNK]-like) provide cellular defense against various stresses. To investigate the role of the MAPK pathways in the toxic effect of beta-toxin, we examined cytotoxicity in five cell lines. Beta-toxin induced cytotoxicity in cells in the following order: THP-1 = U937 > HL-60 > BALL-1 = MOLT-4. In THP-1 cells, beta-toxin formed oligomers on lipid rafts in membranes and induced the efflux of K+ from THP-1 cells in a dose- and time-dependent manner. The phosphorylation of p38 MAPK and JNK occurred in response to an attack by beta-toxin. p38 MAPK (SB203580) and JNK (SP600125) inhibitors enhanced toxin-induced cell death. Incubation in K+-free medium intensified p38 MAPK activation and cell death induced by the toxin, while incubation in K+-high medium prevented those effects. While streptolysin O (SLO) reportedly activates p38 MAPK via reactive oxygen species (ROS), we showed that this pathway did not play a major role in p38 phosphorylation in beta-toxin-treated cells. Therefore, we propose that beta-toxin induces activation of the MAPK pathway to promote host cell survival. PMID:23876806

  18. Quantification of cell proliferation and alpha-toxin gene expression of Clostridium perfringens in the development of necrotic enteritis in broiler chickens.

    PubMed

    Si, Weiduo; Gong, Joshua; Han, Yanming; Yu, Hai; Brennan, John; Zhou, Huaijun; Chen, Shu

    2007-11-01

    Cell proliferation and alpha-toxin gene expression of Clostridium perfringens in relation to the development of necrotic enteritis (NE) were investigated. Unlike bacitracin-treated chickens, non-bacitracin-treated birds exhibited typical NE symptoms and reduced growth performance. They also demonstrated increased C. perfringens proliferation and alpha-toxin gene expression that were positively correlated and progressed according to the regression model y = b(0) + b(1)X - b(2)X(2). The average C. perfringens count of 5 log(10) CFU/g in the ileal digesta appears to be a threshold for developing NE with a lesion score of 2. PMID:17827329

  19. Sialidase Production and Genetic Diversity in Clostridium perfringens Type A Isolated from Chicken with Necrotic Enteritis in Brazil.

    PubMed

    Llanco, Luis A; Nakano, Viviane; Avila-Campos, Mario J

    2015-03-01

    The sialidase activity and genetic diversity of 22 Clostridium perfringens strains isolated from chickens with necrotic enteritis were determined. Sialidase activity was detected in 86.4 % of the strains. All C. perfringens showed a high value of similarity (>96 %), and they were grouped into seven clusters clearly separated from the other reference bacterial strains. From these clusters four patterns were defined in accordance with their phenotypic (sialidase production and antibiotic resistance profile) and genotypic (presence of nanI and nanJ genes) characteristics. Our results showed heterogeneity among strains, but they were genotypically similar, and it is suggested further studies are needed to better understand the pathogenesis of necrotic enteritis. PMID:25373329

  20. 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

  1. Abilities of the mCP Agar method and CRENAME alpha toxin-specific real-time PCR assay to detect Clostridium perfringens spores in drinking water.

    PubMed

    Maheux, Andrée F; Bérubé, Eve; Boudreau, Dominique K; Villéger, Romain; Cantin, Philippe; Boissinot, Maurice; Bissonnette, Luc; Bergeron, Michel G

    2013-12-01

    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

  2. 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

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

    PubMed

    Juneja, Vijay K; Marks, Harry; Huang, Lihan; Thippareddi, H

    2011-06-01

    Comparison of Clostridium 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 water bath stabilized at selected temperatures between 10 and 51°C and sampled periodically. For dynamic experiments, the samples were cooled from 54.4 to 27°C and subsequently from 27 to 4°C for different time periods, designated as x and y hours, respectively. The growth models used were based on a model developed by Baranyi and Roberts (1994. A dynamic approach to predicting bacterial growth in food. Int. J. Food Micro. 23, 277-294), which incorporates a constant, referred to as the physiological state constant, q(0). The value of this constant captures the cells' history before the cooling begins. To estimate specific growth rates, data from isothermal experiments were used, from which a secondary model was developed, based on a form of Ratkowsky's 4-parameter equation. The estimated growth kinetics associated with pork and chicken were similar, but growth appeared to be slightly greater in beef; for beef, the maximum specific growth rates estimated from the Ratkowsky curve was about 2.7 log(10) cfu/h, while for the other two species, chicken and pork, the estimate was about 2.2 log(10) cfu/h. Physiological state constants were estimated by minimizing the mean square error of predictions of the log(10) of the relative increase versus the corresponding observed quantities for the dynamic experiments: for beef the estimate was 0.007, while those for pork and chicken the estimates were about 0.014 and 0.011, respectively. For a hypothetical 1.5h cooling from 54°C to 27° and 5h to 4°C, corresponding to USDA-FSIS cooling compliance guidelines, the predicted growth (log(10) of the relative increase) for each species was: 1.29 for beef; 1.07 for chicken and 0.95 log(10) for pork. However, it was noticed that for pork in particular, the model using the derived q(0) had a tendency to over-predict relative growth when the observed amount of relative growth was small, and under-predict the relative growth when the observed amount of relative growth was large. To provide more fail-safe estimate, rather than using the derived value of q(0), a value of 0.04 is recommended for pork. PMID:21511140

  4. Heterologous protection against alpha toxins of Clostridium perfringens and Staphylococcus aureus induced by binding domain recombinant chimeric protein.

    PubMed

    Uppalapati, Siva R; Kingston, Joseph J; Murali, Harishchandra S; Batra, Harsh V

    2014-05-23

    Clostridium perfringens and Staphylococcus aureus are the two important bacteria frequently associated with majority of the soft tissue infections. The severity and progression of the diseases caused by these pathogens are attributed primarily to the alpha toxins they produce. Previously, we synthesized a non-toxic chimeric molecule r-?CS encompassing the binding domains of C. perfringens and S. aureus alpha toxins and demonstrated that the r-?CS hyperimmune polysera reacts with both the native wild type toxins. In the present report, we evaluated efficacy of r-?CS in conferring protection against C. perfringens and S. aureus alpha toxin infections in murine model. Immunization of BALB/c with r-?CS was effective in inducing both high titers of serum anti-r-?CS antibodies after three administrations. Sub-typing the antibody pool revealed high proportions of IgG1 indicating a Th2-polarized immune response. The r-?CS stimulated the proliferation of splenocytes from the immunized mice upon re-induction by the antigen, in vitro. The levels of interleukin-10 increased while TNF-? was found to be downregulated in the r-?CS induced splenocytes. Mice immunized with r-?CS were protected against intramuscular challenge with 5×LD100 doses of C. perfringens and S. aureus alpha toxins with >80% survival, which killed control animals within 48-72h. Passive immunization of mice with anti-r-?CS serum resulted in 50-80% survival. Our results indicate that r-?CS is a remarkable antigen with protective efficacy against alpha toxin mediated C. perfringens and S. aureus soft tissue co-infections. PMID:24699467

  5. Relationship between hemagglutinin and sialidase from Clostridium perfringens CN3870: gel filtration of mutant and reverant activities.

    PubMed Central

    Rood, J I; Wilkinson, R G

    1976-01-01

    Gel filtration of supernatant fluids, from the wild-type Clostridium perfringens, strain CN3870, and several of the mutants and reverants derived from this strain, showed that these mutants failed to product detectable amounts of still produced sialidase III activity. The reverants tested had regained the ability to produce approximately wild-type levels of the I and II forms of both activities. These results showthat there is a direct relationship between the production of the I form and hemagglutinin and sialidase activities and the production of the II form of these biologically active proteins. Models that explain the genetic basis for these results are discussed. PMID:4435

  6. Comparative genomics of four closely related Clostridium perfringens bacteriophages reveals variable evolution among core genes with therapeutic potential

    Microsoft Academic Search

    Brian B Oakley; Eldin Talundzic; Cesar A Morales; Kelli L Hiett; Gregory R Siragusa; Nikolay V Volozhantsev; Bruce S Seal

    2011-01-01

    Background  Because biotechnological uses of bacteriophage gene products as alternatives to conventional antibiotics will require a thorough\\u000a understanding of their genomic context, we sequenced and analyzed the genomes of four closely related phages isolated from\\u000a Clostridium perfringens, an important agricultural and human pathogen.\\u000a \\u000a \\u000a \\u000a \\u000a Results  Phage whole-genome tetra-nucleotide signatures and proteomic tree topologies correlated closely with host phylogeny. Comparisons\\u000a of our phage genomes

  7. Predictive model for growth of Clostridium perfringens during cooling of cooked uncured beef.

    PubMed

    Juneja, Vijay K; Marks, Harry; Thippareddi, Harshavardhan

    2008-02-01

    This paper considers growth models including one based on Baranyi's equations for growth and the other based on the logistic function. Using a common approach for constructing dynamic models for predicting Clostridium perfringens growth in ready-to-eat uncured beef during cooling, there was no appreciable difference between the models' predictions when the population of cells was within the lag or exponential phases of growth. The developed models can be used for designing safe cooling processes; however, the discrepancies between predicted and observed growths obtained in this study, together with discrepancies reported in other papers using the same, or similar methodology as used in this paper, point to a possible inadequacy of the derived models. In particular, the appropriateness of the methodology depends on the appropriateness of using estimated growth kinetics obtained from experiments conducted in isothermal environments for determining coefficients of differential equations that are used for predicting growth in constantly changing (dynamic) environments. The coefficients are interpreted as instantaneous specific rates of change that are independent of prior history. However, there is no known scientific reason that would imply the truth of this assumption. Incorporating a different, less restrictive assumption, allowing for a dependency on the prior history of cells for these kinetic parameters, might lead to models that provide more accurate estimates of growth. For example, a cooling scenario of 54.4-27 degrees C in 1.5h, the average predicted and observed log(10) relative growths were 1.1log(10) and 0.66log(10), respectively, a difference of 0.44log(10,) whereas, when assuming a particular dependency of history, the predicted value was 0.8log(10). More research is needed to characterize the behavior of growth kinetic parameters relative to prior history in dynamic environments. PMID:17993376

  8. 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

    2014-10-24

    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

  9. 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

  10. Clostridium perfringens enterotoxin and Clostridium difficile toxin A/B do not play a role in acute haemorrhagic diarrhoea syndrome in dogs.

    PubMed

    Busch, K; Suchodolski, J S; Kühner, K A; Minamoto, Y; Steiner, J M; Mueller, R S; Hartmann, K; Unterer, S

    2015-03-01

    Although an association between clostridial pathogens and canine idiopathic acute haemorrhagic diarrhoea syndrome (AHDS) has been described, the relevance of those bacteria and their toxins remains unclear. The aim of this study was to evaluate the association between severity of clinical signs and presence of Clostridium perfringens enterotoxin (CPE) and Clostridium difficile toxin A/B (CDT A/B) in faeces of dogs with AHDS. Faecal samples of 54 dogs with idiopathic AHDS were tested by qualitative CPE and CDT A/B ELISA, and PCR was performed to detect enterotoxin genes of C. perfringens (cpe) and toxin B genes of C. difficile (cdt b). Prevalence of cdt b and CDT A/B in dogs with AHDS was 10/54 and 2/54 versus 3/23 and 0/23 in control dogs. Prevalence of cpe was 35/54 in affected versus 9/23 in control dogs. Prevalence of CPE in dogs with AHDS (13/54) was higher compared with control dogs (0/23). No significant difference was detected between CPE-positive and -negative and between cpe-positive and -negative dogs in severity of clinical signs, duration of hospitalisation, mortality rate and selected laboratory parameters. This study suggests that CPE and CDT A/B do not play a role in idiopathic AHDS, are not associated with clinical parameters in affected dogs and cannot be used to predict disease outcome. PMID:25467148

  11. Clostridium perfringens septicemia in a long-beaked common dolphin Delphinus capensis: an etiology of gas bubble accumulation in cetaceans.

    PubMed

    Danil, Kerri; St Leger, Judy A; Dennison, Sophie; Bernaldo de Quirós, Yara; Scadeng, Miriam; Nilson, Erika; Beaulieu, Nicole

    2014-10-16

    An adult female long-beaked common dolphin Delphinus capensis live-stranded in La Jolla, California, USA, on July 30, 2012 and subsequently died on the beach. Computed tomography and magnetic resonance imaging revealed gas bubble accumulation in the vasculature, organ parenchyma, mandibular fat pads, and subdermal sheath as well as a gas-filled cavity within the liver, mild caudal abdominal effusion, and fluid in the uterus. Gross examination confirmed these findings and also identified mild ulcerations on the palate, ventral skin, and flukes, uterine necrosis, and multifocal parenchymal cavitations in the brain. Histological review demonstrated necrosis and round clear spaces interpreted as gas bubbles with associated bacterial rods within the brain, liver, spleen, and lymph nodes. Anaerobic cultures of the lung, spleen, liver, bone marrow, and abdominal fluid yielded Clostridium perfringens, which was further identified as type A via a multiplex PCR assay. The gas composition of sampled bubbles was typical of putrefaction gases, which is consistent with the by-products of C. perfringens, a gas-producing bacterium. Gas bubble formation in marine mammals due to barotrauma, and peri- or postmortem off-gassing of supersaturated tissues and blood has been previously described. This case study concluded that a systemic infection of C. perfringens likely resulted in production of gas and toxins, causing tissue necrosis. PMID:25320031

  12. 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.

  13. The synergistic necrohemorrhagic action of Clostridium perfringens perfringolysin and alpha toxin in the bovine intestine and against bovine endothelial cells

    PubMed Central

    2013-01-01

    Bovine necrohemorrhagic enteritis is a major cause of mortality in veal calves. Clostridium perfringens is considered as the causative agent, but there has been controversy on the toxins responsible for the disease. Recently, it has been demonstrated that a variety of C. perfringens type A strains can induce necrohemorrhagic lesions in a calf intestinal loop assay. These results put forward alpha toxin and perfringolysin as potential causative toxins, since both are produced by all C. perfringens type A strains. The importance of perfringolysin in the pathogenesis of bovine necrohemorrhagic enteritis has not been studied before. Therefore, the objective of the current study was to evaluate the role of perfringolysin in the development of necrohemorrhagic enteritis lesions in calves and its synergism with alpha toxin. A perfringolysin-deficient mutant, an alpha toxin-deficient mutant and a perfringolysin alpha toxin double mutant were less able to induce necrosis in a calf intestinal loop assay as compared to the wild-type strain. Only complementation with both toxins could restore the activity to that of the wild-type. In addition, perfringolysin and alpha toxin had a synergistic cytotoxic effect on bovine endothelial cells. This endothelial cell damage potentially explains why capillary hemorrhages are an initial step in the development of bovine necrohemorrhagic enteritis. Taken together, our results show that perfringolysin acts synergistically with alpha toxin in the development of necrohemorrhagic enteritis in a calf intestinal loop model and we hypothesize that both toxins act by targeting the endothelial cells. PMID:23782465

  14. VIABILITY OF CLOSTRIDIUM PERFRINGENS, ESCHERICHIA COLI, AND LISTERIA MONOCYTOGNES SURVIVING MILD HEAT OR AQUEOUS OZONE TREATMENT ON BEEF FOLLOWED BY HEAT, ALKALI, OR SALT STRESS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The threat of pathogen survival following ozone treatment of meat necessitates careful evaluation of the surviving microorganisms for tolerance to subsequent heat, pH, and NaCl stress. Log reductions in CFU/g of 3-strain cocktails of Clostridium perfringens, Escherichia coli O157:H7, and Listeria m...

  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. Effect of meat ingredients (sodium nitrite and erythorbate) and processing (vacuum storage and packaging atmosphere) on germination and outgrowth of Clostridium perfringens spores in ham during abusive cooling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of nitrite and erythorbate on Clostridium perfringens spore germination and outgrowth in ham during abusive cooling (15 h) was evaluated. Ham was formulated with ground pork, NaNO2 (0, 50, 100, 150 or 200 ppm) and sodium erythorbate (0 or 547 ppm). Ten grams of meat (stored at 5C for 3 or...

  17. 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...

  18. 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)

    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 b...

  19. 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...

  20. 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...

  1. 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...

  2. 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 ...

  3. PREDICTIVE MODEL FOR GROWTH OF CLOSTRIDIUM PERFRINGENS IN ROAST BEEF DURING COOLING AND INHIBITION OF SPORE GERMINATION AND OUTGROWTH BY SALTS OF ORGANIC ACIDS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Germination and outgrowth of Clostridium perfringens spores in roast beef during chilling was studied following simulated cooling schedules normally used in the processed meat industry. Beef top rounds were formulated to contain a marinade (finished product concentrations of salt, 1%; potassium tet...

  4. 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-...

  5. Further Comparison of Temperature Effects on Growth and Survival of Clostridium perfringens Type A Isolates Carrying a Chromosomal or Plasmid-Borne Enterotoxin Gene

    Microsoft Academic Search

    Jihong Li; Bruce A. McClane

    2006-01-01

    Clostridium perfringens type A isolates can carry the enterotoxin gene (cpe) on either their chromosome or a plasmid, but food poisoning isolates usually have a chromosomal cpe gene. This linkage between chromosomal cpe isolates and food poisoning has previously been attributed, at least in part, to better high-temperature survival of chromosomal cpe isolates than of plasmid cpe isolates. In the

  6. Isolation and identification of Clostridium perfringens in the venom and fangs of Loxosceles intermedia (brown spider): enhancement of the dermonecrotic lesion in loxoscelism

    Microsoft Academic Search

    Cristina Leise B. Monteiro; Rosália Rubel; Laura Lúcia Cogo; Oldemir C. Mangili; Waldemiro Gremski; Silvio S. Veiga

    2002-01-01

    Loxoscelism or the envenoming by the brown spiders (Loxosceles genus spiders), may produce extensive dermonecrosis and hemorrhage at the bite site and, eventually, systemic reactions that may be lethal. Isolation and identification of many different bacteria, among them Clostridium perfringens, of great medical importance due to its involvement in dermonecrotizing and systemic conditions, was carried out from the venomous apparatus

  7. Immunopathology and Cytokine Responses in Broiler Chickens Coinfected with Eimeria maxima and Clostridium perfringens Using an Animal Model of Necrotic Enteritis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The incidence of necrotic enteritis (NE) due to Clostridium perfringens (CP) infection in commercial poultry has been increasing at an alarming rate. While pre-exposure of chickens to coccidia infections is believed to be one of the major risk factors leading to NE, the underlying mechanisms of CP ...

  8. X-ray structure of a novel endolysin encoded by episomal phage phiSM101 of Clostridium perfringens.

    PubMed

    Tamai, Eiji; Yoshida, Hiromi; Sekiya, Hiroshi; Nariya, Hirofumi; Miyata, Shigeru; Okabe, Akinobu; Kuwahara, Tomomi; Maki, Jun; Kamitori, Shigehiro

    2014-04-01

    Gram-positive bacteria possess a thick cell wall composed of a mesh polymer of peptidoglycans, which provides physical protection. Endolysins encoded by phages infecting bacteria can hydrolyse peptidoglycans in the bacterial cell wall, killing the host bacteria immediately. The endolysin (Psm) encoded by episomal phage phiSM101 of enterotoxigenic Clostridium perfringens type A strain SM101 exhibits potent lytic activity towards most strains of Clostridium perfringens. Psm has an N-terminal catalytic domain highly homologous to N-acetylmuramidases belonging to the glycoside hydrolase 25 family, and C-terminal tandem repeated bacterial Src homology 3 (SH3_3) domains as the cell wall-binding domain. The X-ray structure of full-length Psm and a catalytic domain of Psm in complex with N-acetylglucosamine were determined to elucidate the catalytic reaction and cell wall recognition mechanisms of Psm. The results showed that Psm may have adopted a neighbouring-group mechanism for the catalytic hydrolysing reaction in which the N-acetyl carbonyl group of the substrate was involved in the formation of an oxazolinium ion intermediate. Based on structural comparisons with other endolysins and a modelling study, we proposed that tandem repeated SH3_3 domains of Psm recognized the peptide side-chains of peptidoglycans to assist the catalytic domain hydrolysing the glycan backbone. PMID:24674022

  9. Comparison of Tn5397 from Clostridium difficile ,T n916 from Enterococcus faecalis and the CW459tet(M) element from Clostridium perfringens shows that they have similar conjugation regions but different insertion and excision modules

    Microsoft Academic Search

    Adam P. Roberts; Priscilla A. Johanesen; Dena Lyras; Peter Mullany; Julian I. Rood

    2001-01-01

    Comparative analysis of the conjugative transposons Tn5397 from Clostridium difficile and Tn916 from Enterococcus faecalis, and the CW459tet(M) element from Clostridium perfringens, has revealed that these tetracycline-resistance elements are closely related. All three elements contain the tet(M) resistance gene and have sequence similarity throughout their central region. However, they have very different integration\\/excision modules. Instead of the int and xis

  10. 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...

  11. 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...

  12. A sporulation factor is involved in the morphological change of Clostridium perfringens biofilms in response to temperature.

    PubMed

    Obana, Nozomu; Nakamura, Kouji; Nomura, Nobuhiko

    2014-04-01

    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

  13. 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

  14. Three-dimensional structure of a putative non-cellulosomal cohesin module from a Clostridium perfringens family 84 glycoside hydrolase.

    PubMed

    Chitayat, Seth; Gregg, Katie; Adams, Jarrett J; Ficko-Blean, Elizabeth; Bayer, Edward A; Boraston, Alisdair B; Smith, Steven P

    2008-01-01

    The genomes of myonecrotic strains of Clostridium perfringens encode a large number of secreted glycoside hydrolases. The activities of these enzymes are consistent with degradation of the mucosal layer of the human gastrointestinal tract, glycosaminoglycans and other cellular glycans found throughout the body. In many cases this is thought to aid in the propagation of the major toxins produced by C. perfringens. One such example is the family 84 glycoside hydrolases, which contains five C. perfringens members (CpGH84A-E), each displaying a unique modular architecture. The smallest and most extensively studied member, CpGH84C, comprises an N-terminal catalytic domain with beta-N-acetylglucosaminidase activity, a family 32 carbohydrate-binding module, a family 82 X-module (X82) of unknown function, and a fibronectin type-III-like module. Here we present the structure of the X82 module from CpGH84C, determined by both NMR spectroscopy and X-ray crystallography. CpGH84C X82 adopts a jell-roll fold comprising two beta-sheets formed by nine beta-strands. CpGH84C X82 displays distant amino acid sequence identity yet close structural similarity to the cohesin modules of cellulolytic anaerobic bacteria. Cohesin modules are responsible for the assembly of numerous hydrolytic enzymes in a cellulose-degrading multi-enzyme complex, termed the cellulosome, through a high-affinity interaction with the calcium-binding dockerin module. A planar surface is located on the face of the CpGH84 X82 structure that corresponds to the dockerin-binding region of cellulolytic cohesin modules and has the approximate dimensions to accommodate a dockerin module. The presence of cohesin-like X82 modules in glycoside hydrolases of C. perfringens is an indication that the formation of novel X82-dockerin mediated multi-enzyme complexes, with potential roles in pathogenesis, is possible. PMID:17999932

  15. 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

  16. Combined effects of hydrostatic pressure, temperature, and pH on the inactivation of spores of Clostridium perfringens type A and Clostridium sporogenes in buffer solutions.

    PubMed

    Paredes-Sabja, D; Gonzalez, M; Sarker, M R; Torres, J A

    2007-08-01

    To develop a spore inactivation strategy, the effect of 15-min hydrostatic pressure treatments (550 and 650 MPa) at 55 and 75 degrees C in citric acid buffer (4.75 and 6.5 pH) on spores of 5 isolates of Clostridium perfringens type A carrying the gene that encodes the C. perfringens enterotoxin (cpe) on the chromosome (C-cpe), 4 isolates carrying the cpe gene on a plasmid (P-cpe), and 2 strains of C. sporogenes were investigated. Treatments at 650 MPa, 75 degrees C and pH 6.5 were moderately effective against spores of P-cpe (approximately 3.7 decimal reduction, DR) and C. sporogenes (approximately 2.1 DR) but not for C-cpe (approximately 1.0 DR) spores. Treatments at pH 4.75 were moderately effective against spores of P-cpe (approximately 3.2 DR) and C. sporogenes (approximately 2.5 DR) but not of C-cpe (approximately 1.2 DR) when combined with 550 MPa at 75 degrees C. However, when pressure was raised to 650 MPa under the same conditions, high inactivation of P-cpe (approximately 5.1 DR) and C. sporogenes (approximately 5.8 DR) spores and moderate inactivation of C-cpe (approximately 2.8 DR) spores were observed. Further advances in high-pressure treatment strategies to inactivate spores of cpe-positive C. perfringens type A and C. sporogenes more efficiently are needed. PMID:17995687

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

    PubMed Central

    Collie, Renee E.; McClane, Bruce A.

    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 the genotypes of 43 cpe-positive C. perfringens isolates obtained from diverse sources. All North American and European food-poisoning isolates examined in this study were found to carry a chromosomal cpe, while all non-food-borne human GI disease isolates characterized in this study were determined to carry their cpe on an episome. Collectively, these results provide the first evidence that distinct subpopulations of cpe-positive C. perfringens isolates may be responsible for C. perfringens type A food poisoning versus CPE-associated non-food-borne human GI diseases. If these putative associations are confirmed in additional surveys, cpe RFLP and PFGE genotyping assays may facilitate the differential diagnosis of food-borne versus non-food-borne CPE-associated human GI illnesses and may also be useful epidemiologic tools for identifying reservoirs or transmission mechanisms for the subpopulations of cpe-positive isolates specifically responsible for CPE-associated food-borne versus non-food-borne human GI diseases. PMID:9431915

  18. Bilateral abscessed orchiepididymitis associated with sepsis caused by Veillonella parvula and Clostridium perfringens: case report and review of the literature.

    PubMed

    Arrosagaray, P M; Salas, C; Morales, M; Correas, M; Barros, J M; Cordon, M L

    1987-08-01

    Veillonella species is a gram-negative coccus which is part of the anaerobic normal flora in the oral cavity, small intestine, upper respiratory tract, vagina, and urinary tract. The role that this organism plays in infection is not well known, and it is generally associated with other bacteria. We present a case of bilateral abscessed orchiepididymitis associated with septicemia due to Veillonella parvula and, later, to Clostridium perfringens, with the development of severe renal insufficiency and septic shock, which resolved favorably with antibiotic therapy, treatment of shock, and hyperbaric oxygen therapy. In reviewing the literature, we have not found any other case of sepsis due to Veillonella sp. associated with urological disorders. PMID:2887584

  19. Bilateral abscessed orchiepididymitis associated with sepsis caused by Veillonella parvula and Clostridium perfringens: case report and review of the literature.

    PubMed Central

    Arrosagaray, P M; Salas, C; Morales, M; Correas, M; Barros, J M; Cordon, M L

    1987-01-01

    Veillonella species is a gram-negative coccus which is part of the anaerobic normal flora in the oral cavity, small intestine, upper respiratory tract, vagina, and urinary tract. The role that this organism plays in infection is not well known, and it is generally associated with other bacteria. We present a case of bilateral abscessed orchiepididymitis associated with septicemia due to Veillonella parvula and, later, to Clostridium perfringens, with the development of severe renal insufficiency and septic shock, which resolved favorably with antibiotic therapy, treatment of shock, and hyperbaric oxygen therapy. In reviewing the literature, we have not found any other case of sepsis due to Veillonella sp. associated with urological disorders. PMID:2887584

  20. Crystallization and preliminary X-ray diffraction studies of ?-toxin (perfringolysin O), a pore-forming cytolysin of Clostridium perfringens

    NASA Astrophysics Data System (ADS)

    Sugahara, Mitsuaki; Sekino-Suzuki, Naoko; Ohno-Iwashita, Yoshiko; Miki, Kunio

    1996-10-01

    ?-Toxin (perfringolysin O), a cholesterol-binding, pore-forming cytolysin of Clostridium perfringens type A was crystallized by the vapor diffusion procedure using polyethyleneglycol 4000 and sodium chloride as precipitants in 2-(cyclohexylamino)ethanesulfonic acid (CHES) buffer at pH 9.5. The diffraction patterns of precession photographs indicated that the crystals belong to the orthorhombic system and the space group C222 1 with unit-cell dimensions of a = 47.7 Å, b = 182.0 Å and c = 175.8 Å. Assuming that the asymmetric unit contains one or two molecules (Mw 52 700), the Vm value is calculated as 3.6 or 1.8 Å 3/dalton, respectively. The crystals diffract X-rays to at least 3 Å resolution and are suitable for high resolution X-ray crystal structure determination.

  1. Identification of a key residue for oligomerisation and pore-formation of Clostridium perfringens NetB.

    PubMed

    Fernandes da Costa, Sérgio P; Savva, Christos G; Bokori-Brown, Monika; Naylor, Claire E; Moss, David S; Basak, Ajit K; Titball, Richard W

    2014-03-01

    Necrotic enteritis toxin B (NetB) is a ?-pore-forming toxin produced by Clostridium perfringens and has been identified as a key virulence factor in the pathogenesis of avian necrotic enteritis, a disease causing significant economic damage to the poultry industry worldwide. In this study, site-directed mutagenesis was used to identify amino acids that play a role in NetB oligomerisation and pore-formation. NetB K41H showed significantly reduced toxicity towards LMH cells and human red blood cells relative to wild type toxin. NetB K41H was unable to oligomerise and form pores in liposomes. These findings suggest that NetB K41H could be developed as a genetic toxoid vaccine to protect against necrotic enteritis. PMID:24625763

  2. Identification of a Key Residue for Oligomerisation and Pore-Formation of Clostridium perfringens NetB

    PubMed Central

    Fernandes da Costa, Sérgio P.; Savva, Christos G.; Bokori-Brown, Monika; Naylor, Claire E.; Moss, David S.; Basak, Ajit K.; Titball, Richard W.

    2014-01-01

    Necrotic enteritis toxin B (NetB) is a ?-pore-forming toxin produced by Clostridium perfringens and has been identified as a key virulence factor in the pathogenesis of avian necrotic enteritis, a disease causing significant economic damage to the poultry industry worldwide. In this study, site-directed mutagenesis was used to identify amino acids that play a role in NetB oligomerisation and pore-formation. NetB K41H showed significantly reduced toxicity towards LMH cells and human red blood cells relative to wild type toxin. NetB K41H was unable to oligomerise and form pores in liposomes. These findings suggest that NetB K41H could be developed as a genetic toxoid vaccine to protect against necrotic enteritis. PMID:24625763

  3. The Sialidases of Clostridium perfringens type D strain CN3718 differ in their properties and sensitivities to inhibitors.

    PubMed

    Li, Jihong; McClane, Bruce A

    2014-03-01

    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

  4. Molecular Characterization of Podoviral Bacteriophages Virulent for Clostridium perfringens and Their Comparison with Members of the Picovirinae

    PubMed Central

    Volozhantsev, Nikolay V.; Oakley, Brian B.; Morales, Cesar A.; Verevkin, Vladimir V.; Bannov, Vasily A.; Krasilnikova, Valentina M.; Popova, Anastasia V.; Zhilenkov, Eugeni L.; Garrish, Johnna K.; Schegg, Kathleen M.; Woolsey, Rebekah; Quilici, David R.; Line, J. Eric; Hiett, Kelli L.; Siragusa, Gregory R.; Svetoch, Edward A.; Seal, Bruce S.

    2012-01-01

    Clostridium perfringens is a Gram-positive, spore-forming anaerobic bacterium responsible for human food-borne disease as well as non-food-borne human, animal and poultry diseases. Because bacteriophages or their gene products could be applied to control bacterial diseases in a species-specific manner, they are potential important alternatives to antibiotics. Consequently, poultry intestinal material, soil, sewage and poultry processing drainage water were screened for virulent bacteriophages that lysed C. perfringens. Two bacteriophages, designated ?CPV4 and ?ZP2, were isolated in the Moscow Region of the Russian Federation while another closely related virus, named ?CP7R, was isolated in the southeastern USA. The viruses were identified as members of the order Caudovirales in the family Podoviridae with short, non-contractile tails of the C1 morphotype. The genomes of the three bacteriophages were 17.972, 18.078 and 18.397 kbp respectively; encoding twenty-six to twenty-eight ORF's with inverted terminal repeats and an average GC content of 34.6%. Structural proteins identified by mass spectrometry in the purified ?CP7R virion included a pre-neck/appendage with putative lyase activity, major head, tail, connector/upper collar, lower collar and a structural protein with putative lysozyme-peptidase activity. All three podoviral bacteriophage genomes encoded a predicted N-acetylmuramoyl-L-alanine amidase and a putative stage V sporulation protein. Each putative amidase contained a predicted bacterial SH3 domain at the C-terminal end of the protein, presumably involved with binding the C. perfringens cell wall. The predicted DNA polymerase type B protein sequences were closely related to other members of the Podoviridae including Bacillus phage ?29. Whole-genome comparisons supported this relationship, but also indicated that the Russian and USA viruses may be unique members of the sub-family Picovirinae. PMID:22666499

  5. In vitro activities of daptomycin, vancomycin, and penicillin against Clostridium difficile, C. perfringens, Finegoldia magna, and Propionibacterium acnes.

    PubMed

    Tyrrell, Kerin L; Citron, Diane M; Warren, Yumi A; Fernandez, Helen T; Merriam, C Vreni; Goldstein, Ellie J C

    2006-08-01

    Daptomycin has in vitro activity against gram-positive anaerobic bacteria, although limited numbers of species have been tested. We studied the in vitro activities of daptomycin, vancomycin, and penicillin against more than 100 strains each of Clostridium difficile, C. perfringens, Finegoldia magna, and Propionibacterium acnes. Daptomycin Etest MICs and results from time-kill studies were determined for selected strains. For 392 of 421 strains (93%), daptomycin was inhibitory at < or =1 microg/ml, including 15 of 16 strains of C. difficile with elevated linezolid MICs of 8 and 16 microg/ml, all 32 strains with moxifloxacin MICs of > or =4 microg/ml, and all 16 strains resistant to clindamycin. Daptomycin MICs were also < or =1 microg/ml for all 16 F. magna strains resistant to clindamycin and all 32 strains resistant to tetracycline. Only one strain, a C. perfringens strain, had a MIC of >2 microg/ml to daptomycin. Eighty-five and 92.5% of the Etest MICs were within 1 dilution of the agar dilution method for all drugs at 24 and 48 h, respectively. In time-kill studies, a C. difficile strain was inhibited by both daptomycin and vancomycin at 1, 2, 4, 8, and 24 h; colony counts were decreased by 2.3 to 2.9 log at 24 h. Vancomycin was not bactericidal for C. perfringens; however, daptomycin showed bactericidal activity as early as 1 h at four and eight times the MIC and at 2 and 4 h at two and four times the MIC. PMID:16870765

  6. 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

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

    PubMed

    Park, Miseon; Rafii, Fatemeh

    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

  8. 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

  9. Mode of binding of RNA polymerase ? subunit to the phased A-tracts upstream of the phospholipase C gene promoter of Clostridium perfringens.

    PubMed

    Katayama, Seiichi; Ishibashi, Kotaro; Gotoh, Kazuyoshi; Nakamura, Daisuke

    2013-10-01

    Three phased A5-6-tracts lie upstream of the promoter of plc encoding the ?-toxin (phospholipase C) of Clostridium perfringens. The ? subunits of C. perfringens RNA polymerase bind directly to the phased A-tracts via the C-terminal domain of the ? subunit (?CTD). To identify the amino acid residues involved in the binding of C. perfringens ? subunits to the phased A-tracts, 27 amino acid residues in C. perfringens ?CTD were substituted with alanine. The affinities of the mutated ? subunits for the phased A-tracts were examined by gel shift assays and surface plasmon resonance (SPR). The SPR analyses revealed that the phased A-tracts themselves facilitated a complex formation between the phased A-tracts and C. perfringens ? subunits [Kd was 6.1 (±0.3) × 10(-8) M], and that Arg261, Asn264, Gly292 and Lys294 in C. perfringens ?CTD were critical for the binding to the phased A-tracts. The topology of these amino acid residues on the predicted structure of C. perfringens ?CTD indicated a contact path with the phased A-tracts that was similar to that of Escherichia coli ?CTD with the upstream (UP) element. On the other hand, SPR analyses at different temperatures (15, 25 and 37 °C) indicated that the affinity of the C. perfringens ? subunits for the phased A-tracts increased in a low-temperature-dependent manner, whereas that of the E. coli ? subunit for the UP element did not. This suggests that the phased A-tracts may not simply be a subset of the UP element, and that they show specific binding activity with the RNA polymerase ? subunit. PMID:23810806

  10. Evidence of chitinase activity within necrotic enteritis-associated subtypes of Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    C. perfringens (Cp) is associated with the necrotic gastrointestinal condition known as necrotic enteritis (NE) in the chicken. rep-PCR subtyping identified subtypes of Cp from the gastrointestinal tracts of broiler chickens afflicted with NE that were distinguished from strains from environmental,...

  11. 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 ...

  12. 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...

  13. 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...

  14. A novel watery diarrhoea caused by the co-infection of neonatal piglets with Clostridium perfringens type A and Escherichia coli (K88, 987P).

    PubMed

    Wang, Xiaocen; Ren, Wenzhi; Nie, Ying; Cheng, Liqing; Tan, Wei; Wang, Chong; Wei, Libin; Zhang, Rui; Yan, Guangmou

    2013-09-01

    In 2011, a novel watery diarrhoea in 1-7 day-old piglets occurred in Changchun, China, characterized by high pathogenicity and mortality. Investigation of clinical signs, examination for viruses, and isolation and identification of bacteria showed that co-infection by Clostridium perfringens type A and Escherichia coli (K88, 987P) was the most likely cause of the disease. Newborn piglets challenged with a mixture of Clostridium perfringens type A and Escherichia coli (K88, 987P) died within 3 days with clinical signs and gross lesions similar to those in the piglets that died in the outbreak. A subsequent study showed that the use in sows of an inactivated vaccine against the two causal bacteria was effective at reducing the incidence of the watery diarrhoea in piglets. Piglets from sows given the inactivated vaccine had a incidence of watery diarrhoea of 8% (14/175), much lower than the 95% (192/201) seen in piglets from control sows. This is the first report of diarrhoea in piglets resulting from co-infection of Clostridium perfringens type A and Escherichia coli (K88, 987P). Further studies are required to better understand the pathogenesis of this disease. PMID:23846029

  15. Dynamic determination of kinetic parameters, computer simulation, and probabilistic analysis of growth of Clostridium perfringens in cooked beef during cooling.

    PubMed

    Huang, Lihan

    2015-02-16

    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 Clostridium perfringens in cooked beef. This methodology was based on simultaneous numerical analysis and optimization of both primary and secondary models using multiple dynamic growth curves obtained under different conditions. Once the models were constructed, the bootstrap method was used to calculate the 95% confidence intervals of kinetic parameters, and a Monte Carlo simulation method was developed to validate the models using the growth curves not previously used in model development. The results showed that the kinetic parameters obtained from this study accurately matched the common characteristics of C. perfringens, with the optimum temperature being 45.3°C. The results also showed that the predicted growth curves matched accurately with experimental observations used in validation. The mean of residuals of the predictions is -0.02logCFU/g, with a standard deviation of only 0.23logCFU/g. For relative growths <1logCFU/g, the residuals of predictions are <0.4logCFU/g. Overall, 74% of the residuals of predictions are <0.2logCFU/g, 7.7% are >0.4logCFU/g, while only 1.5% are >0.8logCFU/g. In addition, the dynamic model also accurately predicted four isothermal growth curves arbitrarily chosen from the literature. Finally, the Monte Carlo simulation was used to provide the probability of >1 and 2logCFU/g relative growths at the end of cooling. The results of this study will provide a new and accurate tool to the food industry and regulatory agencies to assess the safety of cooked beef in the event of cooling deviation. PMID:25500276

  16. Salmonella spp., Vibrio spp., Clostridium perfringens, and Plesiomonas shigelloides in marine and freshwater invertebrates from coastal California ecosystems.

    PubMed

    Miller, W A; Miller, M A; Gardner, I A; Atwill, E R; Byrne, B A; Jang, S; Harris, M; Ames, J; Jessup, D; Paradies, D; Worcester, K; Melli, A; Conrad, P A

    2006-08-01

    The coastal ecosystems of California are highly utilized by humans and animals, but the ecology of fecal bacteria at the land-sea interface is not well understood. This study evaluated the distribution of potentially pathogenic bacteria in invertebrates from linked marine, estuarine, and freshwater ecosystems in central California. A variety of filter-feeding clams, mussels, worms, and crab tissues were selectively cultured for Salmonella spp., Campylobacter spp., Escherichia coli-O157, Clostridium perfringens, Plesiomonas shigelloides, and Vibrio spp. A longitudinal study assessed environmental risk factors for detecting these bacterial species in sentinel mussel batches. Putative risk factors included mussel collection near higher risk areas for livestock or human sewage exposure, adjacent human population density, season, recent precipitation, water temperature, water type, bivalve type, and freshwater outflow exposure. Bacteria detected in invertebrates included Salmonella spp., C. perfringens, P. shigelloides, Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio alginolyticus. Overall, 80% of mussel batches were culture positive for at least one of the bacterial species, although the pathogens Campylobacter, E. coli-O157, and Salmonella were not detected. Many of the same bacterial species were also cultured from upstream estuarine and riverine invertebrates. Exposure to human sewage sources, recent precipitation, and water temperature were significant risk factors for bacterial detection in sentinel mussel batches. These findings are consistent with the hypothesis that filter-feeding invertebrates along the coast concentrate fecal bacteria flowing from land to sea and show that the relationships between anthropogenic effects on coastal ecosystems and the environmental niches of fecal bacteria are complex and dynamic. PMID:16897302

  17. 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

  18. Clostridium perfringens Beta-Toxin Induces Necrostatin-Inhibitable, Calpain-Dependent Necrosis in Primary Porcine Endothelial Cells

    PubMed Central

    Popoff, Michel; D’Herde, Katharina; Christen, Stephan; Posthaus, Horst

    2013-01-01

    Clostridium perfringens ?-toxin (CPB) is a ?-barrel pore-forming toxin and an essential virulence factor of C. perfringens type C strains, which cause fatal hemorrhagic enteritis in animals and humans. We have previously shown that CPB is bound to endothelial cells within the intestine of affected pigs and humans, and that CPB is highly toxic to primary porcine endothelial cells (pEC) in vitro. The objective of the present study was to investigate the type of cell death induced by CPB in these cells, and to study potential host cell mechanisms involved in this process. CPB rapidly induced lactate dehydrogenase (LDH) release, propidium iodide uptake, ATP depletion, potassium efflux, a marked rise in intracellular calcium [Ca2+]i, release of high-mobility group protein B1 (HMGB1), and caused ultrastructural changes characteristic of necrotic cell death. Despite a certain level of caspase-3 activation, no appreciable DNA fragmentation was detected. CPB-induced LDH release and propidium iodide uptake were inhibited by necrostatin-1 and the two dissimilar calpain inhibitors PD150606 and calpeptin. Likewise, inhibition of potassium efflux, chelation of intracellular calcium and treatment of pEC with cyclosporin A also significantly inhibited CPB-induced LDH release. Our results demonstrate that rCPB primarily induces necrotic cell death in pEC, and that necrotic cell death is not merely a passive event caused by toxin-induced membrane disruption, but is propagated by host cell-dependent biochemical pathways activated by the rise in intracellular calcium and inhibitable by necrostatin-1, consistent with the emerging concept of programmed necrosis (“necroptosis”). PMID:23734212

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

    Microsoft Academic Search

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

    2008-01-01

    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

  20. Haemorhagic enterotoxemia by Clostridium perfringens type C and type A in silver foxes.

    PubMed

    Jarosz, ? S; Gradzki, Z; Smiech, A; Kalinowski, M

    2014-01-01

    Type C and type A of C. perfringens were detected in the seat of natural infections in silver foxes characterized by symptoms of haemorrhagic enterotoxemia. In all of the dead foxes characteristic changes were noted in the small intestine and parenchymatous organs. The production of alpha and beta toxins by isolated bacteria was confirmed by the bioassay using white mice and by PCR. The results of the drug sensitivity testing showed that isolated strains were highly susceptible to amoxicillin with clavulanic acid, metronidazole, doxycycline and penicillin with streptomycin. PMID:24724490

  1. Two novel membrane proteins, TcpD and TcpE, are essential for conjugative transfer of pCW3 in Clostridium perfringens.

    PubMed

    Wisniewski, Jessica A; Teng, Wee L; Bannam, Trudi L; Rood, Julian I

    2015-02-15

    The anaerobic pathogen Clostridium perfringens encodes either toxin genes or antibiotic resistance determinants on a unique family of conjugative plasmids that have a novel conjugation region, the tcp locus. Studies of the paradigm conjugative plasmid from C. perfringens, the 47-kb tetracycline resistance plasmid pCW3, have identified several tcp-encoded proteins that are involved in conjugative transfer and form part of the transfer apparatus. In this study, the role of the conserved hypothetical proteins TcpD, TcpE, and TcpJ was examined. Mutation and complementation analyses showed that TcpD and TcpE were essential for the conjugative transfer of pCW3, whereas TcpJ was not required. To analyze the TcpD and TcpE proteins in C. perfringens, functional hemagglutinin (HA)-tagged derivatives were constructed. Western blots showed that TcpD and TcpE localized to the cell envelope fraction independently of the presence of other pCW3-encoded proteins. Finally, examination of the subcellular localization of TcpD and TcpE by immunofluorescence showed that these proteins were concentrated at both poles of C. perfringens donor cells, where they are postulated to form essential components of the multiprotein complex that comprises the transfer apparatus. PMID:25488300

  2. Crystal structure of the phosphate-binding protein (PBP-1) of an ABC-type phosphate transporter from Clostridium perfringens.

    PubMed

    Gonzalez, Daniel; Richez, Magali; Bergonzi, Celine; Chabriere, Eric; Elias, Mikael

    2014-01-01

    Phosphate limitation is an important environmental stress that affects the metabolism of various organisms and, in particular, can trigger the virulence of numerous bacterial pathogens. Clostridium perfringens, a human pathogen, is one of the most common causes of enteritis necroticans, gas gangrene and food poisoning. Here, we focused on the high affinity phosphate-binding protein (PBP-1) of an ABC-type transporter, responsible for cellular phosphate uptake. We report the crystal structure (1.65 Å resolution) of the protein in complex with phosphate. Interestingly, PBP-1 does not form the short, low-barrier hydrogen bond with phosphate that is typical of previously characterized phosphate-binding proteins, but rather a canonical hydrogen bond. In its unique binding configuration, PBP-1 forms an unusually high number of hydrogen bonds (14) with the phosphate anion. Discrimination experiments reveal that PBP-1 is the least selective PBP characterised so far and is able to discriminate phosphate from its close competing anion, arsenate, by ~150-fold. PMID:25338617

  3. Clostridium perfringens phospholipase C induced ROS production and cytotoxicity require PKC, MEK1 and NF?B activation.

    PubMed

    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

  4. A claudin 3 and claudin 4-targeted Clostridium perfringens protoxin is selectively cytotoxic to PSA-producing prostate cancer cells.

    PubMed

    Romanov, Victor; Whyard, Terry C; Waltzer, Wayne C; Gabig, Theodore G

    2014-09-01

    Prostate cancer is the second leading cause of non-cutaneous cancer-related death in males, and effective strategies for treatment of metastatic disease are currently limited. The tight junction proteins, claudin 3 and claudin 4, serve as cell-surface receptors for the pore-forming Clostridium perfringens enterotoxin [CPE]. Most prostate cancer cells overexpress claudin 3 and claudin 4, and claudins are aberrantly distributed over the plasma membrane, making these cells particularly sensitive to cytolysis by CPE. Prostate cancer cells secrete PSA locally that is proteolytically active; however, circulating PSA is inactivated via binding to protease inhibitors. To overcome systemic toxicity of CPE, a modified protoxin was constructed with a tethered ligand attached to the C-terminus connected by a flexible linker containing a PSA-specific protease cleavage site. This engineered protoxin selectively and efficiently lyses PSA-producing prostate cancer cells whereas CLDN3 and CLDN4 positive cells that do not express PSA are resistant to cytolysis. PMID:24952257

  5. Influence of starch source on sporulation and enterotoxin production by Clostridium perfringens type A.

    PubMed

    Labbe, R; Somers, E; Duncan, C

    1976-03-01

    Of 16 different starch preparations tested, Clostridium perfringes NCTC 8798 yielded maximum sporulation and enterotoxin formation when ICN-soluble starch was included in Duncan and Strong sporulation medium. In general soluble starches were better than potato, corn, or arrowroot starch with regard to these two parameters. PMID:180885

  6. Clostridium perfringens as the Cause of Death of a Captive Atlantic Bottlenosed Dolphin (Tursiops truncatus)

    Microsoft Academic Search

    John D. Buck; L. Louise Shepard

    A previously healthy captive fe- male bottlenosed dolphin (Tursiops truncatus) died suddenly. At necropsy, Clostridium per- f ringens was isolated from dorsal muscle, blood, left heart ventricle, thoracic fluid, and abdom- inal fluid. An identical strain was recovered from pool water. A male dolphin in the same pool had inflicted several \\

  7. The Interaction of a Carbohydrate-Binding Module from a Clostridium perfringens N-Acetyl-beta-hexosaminidase with its Carbohydrate Receptor

    SciTech Connect

    Ficko-Blean,E.; Boraston, A.

    2006-01-01

    Clostridium perfringens is a notable colonizer of the human gastrointestinal tract. This bacterium is quite remarkable for a human pathogen by the number of glycoside hydrolases found in its genome. The modularity of these enzymes is striking as is the frequent occurrence of modules having amino acid sequence identity with family 32 carbohydrate-binding modules (CBMs), often referred to as F5/8 domains. Here we report the properties of family 32 CBMs from a C. perfringens N-acetyl-{beta}-hexosaminidase. Macroarray, UV difference, and isothermal titration calorimetry binding studies indicate a preference for the disaccharide LacNAc ({beta}-d-galactosyl-1,4-{beta}-d-N-acetylglucosamine). The molecular details of the interaction of this CBM with galactose, LacNAc, and the type II blood group H-trisaccharide are revealed by x-ray crystallographic studies at resolutions of 1.49, 2.4, and 2.3 Angstroms, respectively.

  8. 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

  9. 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.

    PubMed

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

    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 (NaNO?: 0 ppm, 100 ppm and 200 ppm) in addition to EO at concentrations of 0.0%, 0.78%, 1.56% and 3.125% stored at 25°C for 30 days. The EO extracted by hydrodistillation and analyzed by gas chromatography-mass spectrometry (CG-MS) was tested in vitro using an agar well diffusion method for determination of minimum inhibitory concentration (MIC) on C. perfringens. According to compositional analysis of the winter savory EO, 26 chemical compounds were identified, and the major constituents were thymol (28.99%), p-cymene (12.00%), linalool (11.00%) and carvacrol (10.71%). The results obtained showed that EO applied at a concentration of 1.56%, which was defined as the MIC, exhibited antimicrobial activity against C. perfringens in the in vitro assays, and the transmission electron microscopy (TEM) revealed structural damage and cell lysis of C. perfringens caused by EO treatment. A synergistic effect between NaNO? and EO was observed. In mortadella-type sausages formulated with 100 ppm of NaNO? and EO at all concentrations tested, the population of target microorganisms was reduced (p?0.05) compared to control samples during all storage period. This data suggests the potential combined use of savory EO and minimal amounts of the synthetic additive, NaNO? to control C. perfringens in mortadella, which goes according to current market trends, where consumers are requesting natural products. PMID:21131083

  10. 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

  11. lesion development in a new intestinal loop model indicates the involvement of a shared Clostridium perfringens virulence factor in haemorrhagic enteritis in calves.

    PubMed

    Valgaeren, B; Pardon, B; Goossens, E; Verherstraeten, S; Schauvliege, S; Timbermont, L; Ducatelle, R; Deprez, P; Van Immerseel, F

    2013-07-01

    Clostridium perfringens-associated enterotoxaemia is a fatal disease in fast growing suckler and veal calves. An intestinal loop model was developed to study the pathogenesis of the disease. Loops were injected with stationary and logarithmic C. perfringens cultures with or without, a milk protein-based commercial milk replacer for calves. Isolates tested were from cases of bovine enterotoxaemia and from calves without signs of enterotoxaemia, in addition to netB-positive and -negative isolates from poultry, a type C isolate from piglets and the human isolate JIR325. All isolates induced necrohaemorrhagic lesions in combination with milk replacer, while all control loops (i.e. medium plus milk replacer) remained histologically normal. In addition, time-course experiments were conducted using an isolate from an outbreak of bovine enterotoxaemia. Histological examination showed that the earliest lesion was congestion of the capillaries, starting within 30 min of inoculation. Haemorrhage and mucosal necrosis began at the tips of the villi 3-4 h after bacterial inoculation. These lesions are similar to those observed in natural cases of bovine enterotoxaemia. Therefore, in this model, necrohaemorrhagic lesions can be induced by C. perfringens isolates from diverse origins, suggesting that the lesions may be caused by one or more virulence factors that are shared by these isolates. PMID:23351504

  12. Recombinant expression of two bacteriophage proteins that lyse clostridium perfringens and share identical sequences in the C-terminal cell wall binding domain of the molecules but are dissimilar in their N-terminal domain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium perfringens is a Gram-positive anaerobic spore-forming bacterium capable of producing four major toxins that are responsible for disease symptoms and pathogenesis in a variety of animals, humans and poultry. The organism is the third leading cause of human food-borne bacterial disease a...

  13. Analysis of genetic similarities between Clostridium perfringens isolates isolated from patients with gas gangrene and from hospital environment conducted with the use of the PFGE method.

    PubMed

    Brzychczy-W?och, Monika; Bulanda, Ma?gorzata

    2014-03-01

    The objective of the study was to perform a comparative analysis of genetic similarity, with the use of pulsed field gel electrophoresis (PFGE), of Clostridium perfringens isolates originating from patients with gas gangrene and from the hospital environment. The study encompassed two patients with a clinical and microbiological diagnosis of gas gangrene, who were hospitalized in one of the hospitals of the Ma?opolska province in the time period between 31st March 2012 and 18th May 2012. Clostridium perfringens isolates genotyping indicated that the isolates originating from the two studied patients did not display genetic similarity and represented two different PFGE types, which corresponded to two different clones (clone A and B). Whereas the strains isolated from the hospital environment were genetically identical with the strain coming from the second patient and represented one PFGE type, which corresponded to one clone (clone A). As a result of the study, it is possible to conclude that both patients developed endogenous infection. Even so, the examination of the hospital environment indicates the possibility of the appearance of exogenous infections. It prompts recommending and following the exact regulations of sanitary regime in the ward and the operating theater if a patient is diagnosed with gas gangrene. PMID:24791817

  14. B-cell epitope of beta toxin of Clostridium perfringens genetically conjugated to a carrier protein: expression, purification and characterization of the chimeric protein.

    PubMed

    Bhatia, Bharti; Solanki, Amit Kumar; Kaushik, Himani; Dixit, Aparna; Garg, Lalit C

    2014-10-01

    Beta toxin (btx) is the prime virulence factor for the pathogenesis of Clostridium perfringens type C strain, known to cause necrotic enteritis and enterotoxaemia in mammalian species. The existing vaccines targeting btx are formaldehyde inactivated culture filtrates of Clostridium. These filtrates raise antigenic load in the host leading to nonspecific and poor responses. The present study aimed to overcome these drawbacks and generate a chimeric protein carrying in silico identified B-cell epitope of btx fused with a carrier protein as a vaccine candidate. Using bioinformatic tools, three stretches of amino acids were predicted as putative B-cell epitopes. One of the epitopes spanning 140-156 amino acid residues was genetically conjugated with B-subunit of heat labile enterotoxin (LTB) of Escherichia coli and expressed as a translational fusion in Vibrio cholerae secretory expression system. High level expression of the recombinant fusion protein rLTB-Btx140-156 was obtained and the protein was successfully purified. The recombinant protein retained the native LTB property to pentamerize and bind to GM1 ganglioside receptor of LTB. The antigenicity of both the epitope and the carrier protein was maintained in fusion protein as indicated by immunoblotting against anti-LTB and anti-btx antibody. The rLTB-Btx140-156 fusion protein therefore can be evaluated as a potential vaccine candidate against C. perfringens. PMID:24996028

  15. The Clostridium perfringens germinant receptor protein GerKC is located in the spore inner membrane and is crucial for spore germination.

    PubMed

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

    2013-11-01

    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 Ca(2+) 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

  16. Differential outgrowth potential of Clostridium perfringens food-borne isolates with various cpe-genotypes in vacuum-packed ground beef during storage at 12°C.

    PubMed

    Xiao, Yinghua; Wagendorp, Arjen; Abee, Tjakko; Wells-Bennik, Marjon H J

    2015-02-01

    In the current study, the outgrowth of spores of 15 different food isolates of Clostridium perfringens was evaluated in vacuum-packed ground beef during storage at 12°C and 25°C. This included enterotoxic strains carrying the gene encoding the CPE enterotoxin on the chromosome (C-cpe), on a plasmid (P-cpe) and cpe-negative strains. The 15 strains were selected from a larger group of strains that were first evaluated for their ability to sporulate in modified Duncan-Strong sporulating medium. Sporulation ability varied greatly between strains but was not associated with a particular cpe genotype. In line with previous studies, the tested C-cpe strains produced spores with significantly higher heat resistance than the cpe-negative and P-cpe strains (both IS1151 and IS1470-like) with the exception of strain VWA009. Following inoculation of vacuum-packed cooked ground beef with spores, the heat-resistant C-cpe strains showed lower outgrowth potential in this model food stored at 12°C than the P-cpe and cpe-negative strains, while no significant differences were observed at 25°C. These results suggest that the latter strains may have a competitive advantage over C-cpe strains at reduced temperatures during storage of foods that support the growth of C. perfringens. While spores of P-cpe strains are readily inactivated by heat processing, post-processing contamination by food handlers who may carry P-cpe strains that have a better growth potential at lower temperatures must be avoided. The varying responses of C. perfringens spores to heat and the differences in outgrowth capacity at different temperatures are factors to be considered in strain selection for challenge tests, and for predictive modelling of C. perfringens. PMID:25461607

  17. Effects of Clostridium perfringens enterotoxin via claudin-4 on normal human pancreatic duct epithelial cells and cancer cells.

    PubMed

    Yamaguchi, Hiroshi; Kojima, Takashi; Ito, Tatsuya; Kyuno, Daisuke; Kimura, Yasutoshi; Imamura, Masafumi; Hirata, Koichi; Sawada, Norimasa

    2011-09-01

    The tight junction protein claudin-4 is frequently overexpressed in pancreatic cancer, and is also a receptor for Clostridium perfringens enterotoxin (CPE). The cytotoxic effects of CPE are thought to be useful as a novel therapeutic tool for pancreatic cancer. However, the responses to CPE via claudin-4 remain unknown in normal human pancreatic duct epithelial (HPDE) cells. We introduced the human telomerase reverse transcriptase (hTERT) gene into HPDE cells in primary culture as a model of normal HPDE cells in vitro. hTERT-HPDE cells treated with or without 10% FBS and pancreatic cancer cell lines PANC-1, BXPC3, HPAF-II and HPAC were treated with CPE. In Western blotting, the expression of claudin-4 protein in hTERT-HPDE cells treated with 10% FBS was as high as it was in all of the pancreatic cancer cell lines. In hTERT-HPDE cells with or without 10% FBS, cytotoxicity was not observed at any concentration of CPE, whereas in all pancreatic cancer cell lines, CPE had a dose-dependent cytotoxic effect. In hTERT-HPDE cells with 10% FBS, claudin-4 was localized in the apical-most regions, where there are tight junction areas, in which in all pancreatic cancer cell lines claudin-4 was found not only in the apical-most regions but also at basolateral membranes. In hTERT-HPDE cells with 10% FBS after treatment with CPE, downregulation of barrier function and claudin-4 expression at the membranes was observed. In HPAC cells, the sensitivity to CPE was significantly decreased by knockdown of claudin-4 expression using siRNA compared to the control. These findings suggest that, in normal HPDE cells, the lack of toxicity of CPE was probably due to the localization of claudin-4, which is different from that of pancreatic cancer cells. hTERT-HPDE cells in this culture system may be a useful model of normal HPDE cells not only for physiological regulation of claudin-4 expression but also for developing safer and more effective therapeutic methods targeting claudin-4 in pancreatic cancer. PMID:21573709

  18. Effects of Dietary Additives and Early Feeding on Performance, Gut Development and Immune Status of Broiler Chickens Challenged with Clostridium perfringens

    PubMed Central

    Ao, Z.; Kocher, A.; Choct, M.

    2012-01-01

    The effects of dietary additives and holding time on resistance and resilience of broiler chickens to Clostridium perfringens challenge were investigated by offering four dietary treatments. These were a negative control (basal), a positive control (Zn-bacitracin) and two dietary additives, mannanoligosaccharides (MOS), and acidifier. Two holding times included (a) immediate access to feed and water post hatch (FED) and (b) access to both feed and water 48 h post hatch (HELD). Chicks fed Zn-bacitracin had no intestinal lesions attributed to necrotic enteritis (NE), whereas chicks fed both MOS or acidifier showed signs of NE related lesions. All dietary treatments were effective in reducing the numbers of C. perfringens in the ileum post challenge. The FED chicks had heavier body weight and numerically lower mortality. The FED chicks also showed stronger immune responses to NE challenge, showing enhanced (p<0.05) proliferation of T-cells. Early feeding of the MOS supplemented diet increased (p<0.05) IL-6 production. The relative bursa weight of the FED chicks was heavier at d 21 (p<0.05). All the additives increased the relative spleen weight of the HELD chicks at d 14 (p<0.05). The FED chicks had increased villus height and reduced crypt depth, and hence an increased villus/crypt ratio, especially in the jejunum at d 14 (p<0.05). The same was true for the HELD chicks given dietary additives (p<0.05). It may be concluded that the chicks with early access to dietary additives showed enhanced immune response and gut development, under C. perfringens challenge. The findings of this study shed light on managerial and nutritional strategies that could be used to prevent NE in the broiler industry without the use of in-feed antibiotics. PMID:25049595

  19. Effects of Dietary Additives and Early Feeding on Performance, Gut Development and Immune Status of Broiler Chickens Challenged with Clostridium perfringens.

    PubMed

    Ao, Z; Kocher, A; Choct, M

    2012-04-01

    The effects of dietary additives and holding time on resistance and resilience of broiler chickens to Clostridium perfringens challenge were investigated by offering four dietary treatments. These were a negative control (basal), a positive control (Zn-bacitracin) and two dietary additives, mannanoligosaccharides (MOS), and acidifier. Two holding times included (a) immediate access to feed and water post hatch (FED) and (b) access to both feed and water 48 h post hatch (HELD). Chicks fed Zn-bacitracin had no intestinal lesions attributed to necrotic enteritis (NE), whereas chicks fed both MOS or acidifier showed signs of NE related lesions. All dietary treatments were effective in reducing the numbers of C. perfringens in the ileum post challenge. The FED chicks had heavier body weight and numerically lower mortality. The FED chicks also showed stronger immune responses to NE challenge, showing enhanced (p<0.05) proliferation of T-cells. Early feeding of the MOS supplemented diet increased (p<0.05) IL-6 production. The relative bursa weight of the FED chicks was heavier at d 21 (p<0.05). All the additives increased the relative spleen weight of the HELD chicks at d 14 (p<0.05). The FED chicks had increased villus height and reduced crypt depth, and hence an increased villus/crypt ratio, especially in the jejunum at d 14 (p<0.05). The same was true for the HELD chicks given dietary additives (p<0.05). It may be concluded that the chicks with early access to dietary additives showed enhanced immune response and gut development, under C. perfringens challenge. The findings of this study shed light on managerial and nutritional strategies that could be used to prevent NE in the broiler industry without the use of in-feed antibiotics. PMID:25049595

  20. Further comparison of temperature effects on growth and survival of Clostridium perfringens type A isolates carrying a chromosomal or plasmid-borne enterotoxin gene.

    PubMed

    Li, Jihong; McClane, Bruce A

    2006-07-01

    Clostridium perfringens type A isolates can carry the enterotoxin gene (cpe) on either their chromosome or a plasmid, but food poisoning isolates usually have a chromosomal cpe gene. This linkage between chromosomal cpe isolates and food poisoning has previously been attributed, at least in part, to better high-temperature survival of chromosomal cpe isolates than of plasmid cpe isolates. In the current study we assessed whether vegetative cells and spores of chromosomal cpe isolates also survive better than vegetative cells and spores of plasmid cpe isolates survive when the vegetative cells and spores are subjected to low temperatures. Vegetative cells of chromosomal cpe isolates exhibited about eightfold-higher decimal reduction values (D values) at 4 degrees C and threefold-higher D values at -20 degrees C than vegetative cells of plasmid cpe isolates exhibited. After 6 months of incubation at 4 degrees C and -20 degrees C, the average log reductions in viability for spores of plasmid cpe isolates were about fourfold and about threefold greater, respectively, than the average log reductions in viability for spores from chromosomal cpe isolates. C. perfringens type A isolates carrying a chromosomal cpe gene also grew significantly faster than plasmid cpe isolates grew at 25 degrees C, 37 degrees C, or 43 degrees C. In addition, chromosomal cpe isolates grew at higher maximum and lower minimum temperatures than plasmid cpe isolates grew. Collectively, these results suggest that chromosomal cpe isolates are commonly involved in food poisoning because of their greater resistance to low (as well as high) temperatures for both survival and growth. They also indicate the importance of proper low-temperature storage conditions, as well as heating, for prevention of C. perfringens type A food poisoning. PMID:16820444

  1. Relative disease susceptibility and clostridial toxin antibody responses in three commercial broiler lines coinfected with Clostridium perfringens and Eimeria maxima using an experimental model of necrotic enteritis.

    PubMed

    Jang, Seung I; Lillehoj, Hyun S; Lee, Sung-Hyen; Lee, Kyung Woo; Lillehoj, Erik P; Hong, Yeong Ho; An, Dong-Jun; Jeoung, D Hye-Young; Chun, Ji-Eun

    2013-09-01

    Necrotic enteritis is an enteric disease of poultry resulting from infection by Clostridium perfringens with coinfection by Eimeria spp. constituting a major risk factor for disease pathogenesis. This study compared three commercial broiler chicken lines using an experimental model of necrotic enteritis. Day-old male Cobb, Ross, and Hubbard broilers were orally infected with viable C. perfringens and E. maxima and fed a high-protein diet to promote the development of experimental disease. Body weight loss, intestinal lesions, and serum antibody levels against alpha-toxin and necrotic enteritis B-like (NetB) toxin were measured as parameters of disease susceptibility and host immune response. Cobb chickens exhibited increased body weight loss compared with Ross and Hubbard breeds and greater gut lesion severity compared with Ross chickens. NetB antibody levels were greater in Cobb chickens compared with the Ross or Hubbard groups. These results suggest that Cobb chickens may be more susceptible to necrotic enteritis in the field compared with the Ross and Hubbard lines. PMID:24283139

  2. Immunization with recombinant bivalent chimera r-Cpae confers protection against alpha toxin and enterotoxin of Clostridium perfringens type A in murine model.

    PubMed

    Shreya, Das; Uppalapati, Siva R; Kingston, Joseph J; Sripathy, Murali H; Batra, Harsh V

    2015-05-01

    Clostridium perfringens type A, an anaerobic pathogen is the most potent cause of soft tissue infections like gas gangrene and enteric diseases like food poisoning and enteritis. The disease manifestations are mediated via two important exotoxins, viz. myonecrotic alpha toxin (?C) and enterotoxin (CPE). In the present study, we synthesized a bivalent chimeric protein r-Cpae comprising C-terminal binding regions of ?C and CPE using structural vaccinology rationale and assessed its protective efficacy against both alpha toxin (?C) and enterotoxin (CPE) respectively, in murine model. Active immunization of mice with r-Cpae generated high circulating serum IgG (systemic), significantly increased intestinal mucosal s-IgA antibody titres and resulted in substantial protection to the immunized animals (100% and 75% survival) with reduced tissue morbidity when administered with 5×LD100 doses of ?C (intramuscular) and CPE (intra-gastric gavage) respectively. Mouse RBCs and Caco-2 cells incubated with a mixture of anti-r-Cpae antibodies and ?C and CPE respectively, illustrated significantly higher protection against the respective toxins. Passive immunization of mice with a similar mixture resulted in 91-100% survival at the end of the 15 days observation period while mice immunized with a concoction of sham sera and respective toxins died within 2-3 days. This work demonstrates the efficacy of the rationally designed r-Cpae chimeric protein as a potential sub unit vaccine candidate against ?C and CPE of C. perfringens type A toxemia. PMID:25645504

  3. [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

  4. 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?

    PubMed Central

    Zekarias, Bereket; Mo, Hua; Curtiss, Roy

    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 antimicrobial-free farming. The pathogenesis of NE is associated with the proliferation of C. perfringens in the small intestine and the secretion of large amounts of alpha toxin, the major virulence factor. Since there is no vaccine for NE, we have developed a candidate live oral recombinant attenuated Salmonella enterica serovar Typhimurium vaccine (RASV) that delivers a nontoxic fragment of alpha toxin. The 3? end of the plc gene, encoding the C-terminal domain of alpha toxin (PlcC), was cloned into plasmids that enable the expression and secretion of PlcC fused to a signal peptide. Plasmids were inserted into Salmonella enterica serovar Typhimurium host strain ?8914, which has attenuating pabA and pabB deletion mutations. Three-day-old broiler chicks were orally immunized with 109 CFU of the vaccine strain and developed alpha toxin-neutralizing serum antibodies. When serum from these chickens was added into C. perfringens broth cultures, bacterial growth was suppressed. In addition, immunofluorescent microscopy showed that serum antibodies bind to the bacterial surface. The immunoglobulin G (IgG) and IgA titers in RASV-immunized chickens were low; however, when the chickens were given a parenteral boost injection with a purified recombinant PlcC protein (rPlcC), the RASV-immunized chickens mounted rapid high serum IgG and bile IgA titers exceeding those primed by rPlcC injection. RASV-immunized chickens had reduced intestinal mucosal pathology after challenge with virulent C. perfringens. These results indicate that oral RASV expressing an alpha toxin C-terminal peptide induces protective immunity against NE. PMID:18337376

  5. 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 ...

  6. Contributions of NanI sialidase to Caco-2 cell adherence by Clostridium perfringens type A and C strains causing human intestinal disease.

    PubMed

    Li, Jihong; McClane, Bruce A

    2014-11-01

    Previous studies showed that Clostridium perfringens type D animal disease strain CN3718 uses NanI sialidase for adhering to enterocyte-like Caco-2 cells. The current study analyzed whether NanI is similarly important when type A and C human intestinal disease strains attach to Caco-2 cells. A PCR survey determined that the nanI gene was absent from typical type A food poisoning (FP) strains carrying a chromosomal enterotoxin (CPE) gene or the genetically related type C Darmbrand (Db) strains. However, the nanI gene was present in type A strains from healthy humans, type A strains causing CPE-associated antibiotic-associated diarrhea (AAD) or sporadic diarrhea (SD), and type C Pig-Bel strains. Consistent with NanI sialidase being the major C. perfringens sialidase when produced, FP and Db strains had little supernatant sialidase activity compared to other type A or C human intestinal strains. All type A and C human intestinal strains bound to Caco-2 cells, but NanI-producing strains had higher attachment levels. When produced, NanI can contribute to host cell attachment of human intestinal disease strains, since a nanI null mutant constructed in type A SD strain F4969 had lower Caco-2 cell adhesion than wild-type F4969 or a complemented strain. Further supporting a role for NanI in host cell attachment, sialidase inhibitors reduced F4969 adhesion to Caco-2 cells. Collectively, these results suggest that NanI may contribute to the intestinal attachment and colonization needed for the chronic diarrhea of CPE-associated AAD and SD, but this sialidase appears to be dispensable for the acute pathogenesis of type A FP or type C enteritis necroticans. PMID:25135687

  7. A Novel Pore-Forming Toxin in Type A Clostridium perfringens Is Associated with Both Fatal Canine Hemorrhagic Gastroenteritis and Fatal Foal Necrotizing Enterocolitis

    PubMed Central

    Nowell, Victoria J.; Nicholson, Vivian M.; Oliphant, Kaitlyn; Prescott, John F.

    2015-01-01

    A role for type A Clostridium perfringens in acute hemorrhagic and necrotizing gastroenteritis in dogs and in necrotizing enterocolitis of neonatal foals has long been suspected but incompletely characterized. The supernatants of an isolate made from a dog and from a foal that died from these diseases were both found to be highly cytotoxic for an equine ovarian (EO) cell line. Partial genome sequencing of the canine isolate revealed three novel putative toxin genes encoding proteins related to the pore-forming Leukocidin/Hemolysin Superfamily; these were designated netE, netF, and netG. netE and netF were located on one large conjugative plasmid, and netG was located with a cpe enterotoxin gene on a second large conjugative plasmid. Mutation and complementation showed that only netF was associated with the cytotoxicity. Although netE and netG were not associated with cytotoxicity, immunoblotting with specific antisera showed these proteins to be expressed in vitro. There was a highly significant association between the presence of netF with type A strains isolated from cases of canine acute hemorrhagic gastroenteritis and foal necrotizing enterocolitis. netE and netF were found in all cytotoxic isolates, as was cpe, but netG was less consistently present. Pulsed-field gel electrophoresis showed that netF-positive isolates belonged to a clonal population; some canine and equine netF-positive isolates were genetically indistinguishable. Equine antisera to recombinant Net proteins showed that only antiserum to rNetF had high supernatant cytotoxin neutralizing activity. The identifica-tion of this novel necrotizing toxin is an important advance in understanding the virulence of type A C. perfringens in specific enteric disease of animals. PMID:25853427

  8. Immunological responses to Clostridium perfringens alpha-toxin in two genetically divergent lines of chickens as influenced by major histocompatibility complex genotype.

    PubMed

    Sumners, L H; Cox, C M; Kim, S; Salevsky, J E; Siegel, P B; Dalloul, R A

    2012-03-01

    Chickens genetically selected for low (LA) or high (HA) antibody response to SRBC displayed a correlated change in MHC, so that LA chickens were 96% B13 and HA chickens were 96% B21. The LA line appears to be less susceptible to invasion by extracellular pathogens, whereas HA chickens are more resistant to infection by intracellular organisms. Resistance to Clostridium perfringens is one instance in which the lines do not follow their established trend of pathogen susceptibility, where during a clinical outbreak of necrotic enteritis, B21B21 genotypes experienced significantly less mortality than B13B13 genotypes. A study was carried out to assess immunological differences between LA and HA lines during exposure to C. perfringens ?-toxin. Peripheral blood mononuclear cells were isolated from each genetic line, cultured with or without lipopolysaccharide (4 h), and exposed to varying concentrations of ?-toxin (1; 10; 100; and 1,000 U/L) for 2 and 4 h. Evaluation of cellular proliferation, percentage of cytotoxicity, and immunological gene expression was carried out in a series of experiments. Cells isolated from HA chickens had significantly increased proliferation than those from LA chickens at low toxin levels (1 and 10 U/L) and significantly decreased proliferation at high toxin levels (100 and 1,000 U/L). Following exposure to lipopolysaccharide, the percentage of cytotoxicity was higher for LA than HA cells. In both assays, HA cells displayed superior performance following lipopolysaccharide-stimulation. Gene expression analysis of immune transcripts by quantitative real-time PCR revealed significantly upregulated expression of interferon (IFN)-?, interleukin (IL)-8, IL-13 (2 h), IL-15, and CXCLi1 (4 h) in HA than LA chickens. Cells isolated from the LA line displayed significantly elevated expression of IL-2, IL-10, IL-13 (4 h), IL-16, IL-18, inducible nitric oxide synthase (iNOS), CXCLi1 (2 h), and lipopolysaccharide-induced tumor necrosis factor-? factor (LITAF) compared with the HA line. Clearly, these 2 genetic lines display highly divergent immune responses in regards to C. perfringens toxin exposure. PMID:22334734

  9. 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

  10. Release of glycoprotein (GP1) from the tegumental surface of Taenia solium by phospholipase C from Clostridium perfringens suggests a novel protein-anchor to membranes.

    PubMed

    Landa, Abraham; Willms, Kaethe; Laclette, Juan Pedro

    2010-01-01

    In order to explore how molecules are linked to the membrane surface in larval Taenia solium, whole cysticerci were incubated in the presence of phospholipase C from Clostridium perfringens (PLC). Released material was collected and analyzed in polyacrylamide gels with sodium dodecyl sulfate. Two major bands with apparent molecular weights of 180 and 43 kDa were observed. Western blot of released material and localization assays in cysticerci tissue sections using antibodies against five known surface glycoproteins of T. solium cysticerci indicated that only one, previously called GP1, was released. Similar localization studies using the lectins wheat-germ-agglutinin and Concanavalin A showed that N-acetyl-D-glucosamine, N-acetylneuraminic, sialic acid, alphamethyl-D-mannoside, D-manose/glucose, and N-acetyl-D-glucosamine residues are abundantly present on the surface. On the other hand, we find that treatment with PLC releases molecules from the surface; they do not reveal Cross Reacting Determinant (CRD), suggesting a novel anchor to the membrane for the glycoprotein GP1. PMID:20130782

  11. Release of Glycoprotein (GP1) from the Tegumental Surface of Taenia solium by Phospholipase C from Clostridium perfringens Suggests a Novel Protein-Anchor to Membranes

    PubMed Central

    Landa, Abraham; Willms, Kaethe; Laclette, Juan Pedro

    2010-01-01

    In order to explore how molecules are linked to the membrane surface in larval Taenia solium, whole cysticerci were incubated in the presence of phospholipase C from Clostridium perfringens (PLC). Released material was collected and analyzed in polyacrylamide gels with sodium dodecyl sulfate. Two major bands with apparent molecular weights of 180 and 43?kDa were observed. Western blot of released material and localization assays in cysticerci tissue sections using antibodies against five known surface glycoproteins of T. solium cysticerci indicated that only one, previously called GP1, was released. Similar localization studies using the lectins wheat-germ-agglutinin and Concanavalin A showed that N-acetyl-D-glucosamine, N-acetylneuraminic, sialic acid, ?methyl-D-mannoside, D-manose/glucose, and N-acetyl-D-glucosamine residues are abundantly present on the surface. On the other hand, we find that treatment with PLC releases molecules from the surface; they do not reveal Cross Reacting Determinant (CRD), suggesting a novel anchor to the membrane for the glycoprotein GP1. PMID:20130782

  12. Recombinant Expression of Two Bacteriophage Proteins That Lyse Clostridium perfringens and Share Identical Sequences in the C-Terminal Cell Wall Binding Domain of the Molecules but Are Dissimilar in Their N-Terminal Active Domains

    PubMed Central

    Simmons, Mustafa; Donovan, David M.; Siragusa, Gregory R.; Seal, Bruce S.

    2014-01-01

    Clostridium perfringens is a Gram-positive anaerobic spore-forming bacterium capable of producing four major toxins that are responsible for disease symptoms and pathogenesis in a variety of animals, humans, and poultry. The organism is the third leading cause of human foodborne bacterial disease, and C. perfringens is the presumptive etiologic agent of necrotic enteritis among chickens, which in the acute form can cause increased mortality among broiler flocks. Countries that have complied with the ban on antimicrobial growth promoters (AGP) in feeds have had increased incidences of C. perfringens-associated necrotic enteritis in poultry. To address this issue, new antimicrobial agents, putative lysins from the genomes of bacteriophages, are identified. Two putative phage lysin genes (ply) from the clostridial phages phiCP39O and phiCP26F were cloned and expressed in Escherichia coli, and the resultant proteins were purified to near homogeneity. Gene and protein sequencing revealed that the predicted and chemically determined amino acid sequences of the two recombinant proteins were homologous to N-acetylmuramoyl-L-alanine amidases. The proteins were identical in the C-terminal putative cell-wall binding domain, but only 55% identical to each other in the presumptive N-terminal catalytic domain. Both recombinant lysins were capable of lysing both parental phage host strains of C. perfringens as well as other strains of the bacterium in spot and turbidity reduction assays. The observed reduction in turbidity was correlated with up to a 3 log cfu/mL reduction in viable C. perfringens on brain–heart infusion agar plates. However, other member species of the clostridia were resistant to the lytic activity by both assays. PMID:20825156

  13. 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

  14. Gene expression profiling within the spleen of Clostridium perfringens-challenged Broilers fed antibiotic-medicated and non-medicated diets

    PubMed Central

    Sarson, Aimie J; Wang, Ying; Kang, Zhumei; Dowd, Scot E; Lu, Yang; Yu, Hai; Han, Yanming; Zhou, Huaijun; Gong, Joshua

    2009-01-01

    Background Clostridium perfringens (Cp) is a Gram-positive anaerobic bacterium that causes necrotic enteritis (NE) in poultry when it overgrows in the small intestine. NE disease has previously been controlled through the use of growth-promoting antibiotics. This practice was recently banned in European countries, leading to significantly increased incidence of NE threatening the poultry industry. Control strategies and technology as substitutes to dietary antibiotics are therefore urgently required. To develop the substitutes, it is important to understand host immune responses to Cp infection. However, the knowledge is still lacking. We therefore investigated gene expression profiles within immunologically-relevant tissue, the spleen, in order to identify factors that are involved in immunity to NE and have potential as therapeutic targets. Results Use of a 44 K Agilent chicken genome microarray revealed significant up-regulation of many immune-associated genes in Cp-challenged chickens, including galectin 3, IFNAR1, IgY-receptor, TCR?, granzyme A, and mannose-6-P-R, which were subsequently validated by quantitative PCR assays. Functional annotation of differentially expressed genes was conducted using the High Throughput Gene Ontology Functional Annotation database. Medicated and Non-medicated chickens had similar annotation profiles with cell activities and regulation being the most dominant biological processes following Cp infection. Conclusion Broiler chickens demonstrated an intricate and holistic magnitude of host response to Cp challenge and the development of NE. Although the influence of dietary antibiotics appeared to be less significant than the disease process, both had a considerable impact on the host response. Markers previously identified in intestinal inflammatory diseases of other species, including humans, and indicators of enhanced antibody responses, appeared to be involved in the chicken response to Cp challenge. The significance in host immune responses of immune mediators identified from the present study warrants further studies to verify their functions during NE development and to determine their potential application to control NE disease. PMID:19500416

  15. Cysteine-Scanning Mutagenesis Supports the Importance of Clostridium perfringens Enterotoxin Amino Acids 80 to 106 for Membrane Insertion and Pore Formation

    PubMed Central

    Chen, Jianwu; Theoret, James R.; Shrestha, Archana; Smedley, James G.

    2012-01-01

    Clostridium perfringens enterotoxin (CPE) causes the gastrointestinal symptoms of the second most common bacterial food-borne illness. Previous studies suggested that a region named TM1, which has amphipathic characteristics and spans from amino acids 81 to 106 of the native CPE protein, forms a ?-hairpin involved in ?-barrel pore formation. To further explore the potential role of TM1 in pore formation, the single Cys naturally present in CPE at residue 186 was first altered to alanine by mutagenesis; the resultant rCPE variant, named C186A, was shown to retain cytotoxic properties. Cys-scanning mutagenesis was then performed in which individual Cys mutations were introduced into each TM1 residue of the C186A variant. When those Cys variants were characterized, three variants were identified that exhibit reduced cytotoxicity despite possessing binding and oligomerization abilities similar to those of the C186A variant from which they were derived. Pronase challenge experiments suggested that the reduced cytotoxicity of those two Cys variants, i.e., the F91C and F95C variants, which model to the tip of the ?-hairpin, was attributable to a lessened ability of these variants to insert into membranes after oligomerization. In contrast, another Cys variant, i.e., the G103C variant, with impaired cytotoxicity apparently inserted into membranes after oligomerization but could not form a pore with a fully functional channel. Collectively, these results support the TM1 region forming a ?-hairpin as an important step in CPE insertion and pore formation. Furthermore, this work identifies the first amino acid residues specifically involved in those two steps in CPE action. PMID:22966051

  16. Mechanism of Clostridium perfringens enterotoxin interaction with claudin-3/-4 protein suggests structural modifications of the toxin to target specific claudins.

    PubMed

    Veshnyakova, Anna; Piontek, Jörg; Protze, Jonas; Waziri, Negar; Heise, Ivonne; Krause, Gerd

    2012-01-13

    Claudins (Cld) are essential constituents of tight junctions. Domain I of Clostridium perfringens enterotoxin (cCPE) binds to the second extracellular loop (ECL2) of a subset of claudins, e.g. Cld3/4 and influences tight junction formation. We aimed to identify interacting interfaces and to alter claudin specificity of cCPE. Mutagenesis, binding assays, and molecular modeling were performed. Mutation-guided ECL2 docking of Cld3/4 onto the crystal structure of cCPE revealed a common orientation of the proposed ECL2 helix-turn-helix motif in the binding cavity of cCPE: residues Leu(150)/Leu(151) of Cld3/4 bind similarly to a hydrophobic pit formed by Tyr(306), Tyr(310), and Tyr(312) of cCPE, and Pro(152)/Ala(153) of Cld3/4 is proposed to bind to a second pit close to Leu(223), Leu(254), and Leu(315). However, sequence variation in ECL2 of these claudins is likely responsible for slightly different conformation in the turn region, which is in line with different cCPE interaction modes of Cld3 and Cld4. Substitutions of other so far not characterized cCPE residues lining the pocket revealed two spatially separated groups of residues (Leu(223), Asp(225), and Arg(227) and Leu(254), lle(258), and Asp(284)), which are involved in binding to Cld3 and Cld4, albeit differently. Involvement of Asn(148) of Cld3 in cCPE binding was confirmed, whereas no evidence for involvement of Lys(156) or Arg(157) was found. We show structure-based alteration of cCPE generating claudin binders, which interact subtype-specific preferentially either with Cld3 or with Cld4. The obtained mutants and mechanistic insights will advance the design of cCPE-based modulators to target specific claudin subtypes related either to paracellular barriers that impede drug delivery or to tumors. PMID:22128179

  17. Mechanism of Clostridium perfringens Enterotoxin Interaction with Claudin-3/-4 Protein Suggests Structural Modifications of the Toxin to Target Specific Claudins*

    PubMed Central

    Veshnyakova, Anna; Piontek, Jörg; Protze, Jonas; Waziri, Negar; Heise, Ivonne; Krause, Gerd

    2012-01-01

    Claudins (Cld) are essential constituents of tight junctions. Domain I of Clostridium perfringens enterotoxin (cCPE) binds to the second extracellular loop (ECL2) of a subset of claudins, e.g. Cld3/4 and influences tight junction formation. We aimed to identify interacting interfaces and to alter claudin specificity of cCPE. Mutagenesis, binding assays, and molecular modeling were performed. Mutation-guided ECL2 docking of Cld3/4 onto the crystal structure of cCPE revealed a common orientation of the proposed ECL2 helix-turn-helix motif in the binding cavity of cCPE: residues Leu150/Leu151 of Cld3/4 bind similarly to a hydrophobic pit formed by Tyr306, Tyr310, and Tyr312 of cCPE, and Pro152/Ala153 of Cld3/4 is proposed to bind to a second pit close to Leu223, Leu254, and Leu315. However, sequence variation in ECL2 of these claudins is likely responsible for slightly different conformation in the turn region, which is in line with different cCPE interaction modes of Cld3 and Cld4. Substitutions of other so far not characterized cCPE residues lining the pocket revealed two spatially separated groups of residues (Leu223, Asp225, and Arg227 and Leu254, lle258, and Asp284), which are involved in binding to Cld3 and Cld4, albeit differently. Involvement of Asn148 of Cld3 in cCPE binding was confirmed, whereas no evidence for involvement of Lys156 or Arg157 was found. We show structure-based alteration of cCPE generating claudin binders, which interact subtype-specific preferentially either with Cld3 or with Cld4. The obtained mutants and mechanistic insights will advance the design of cCPE-based modulators to target specific claudin subtypes related either to paracellular barriers that impede drug delivery or to tumors. PMID:22128179

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

    PubMed

    Lund, Barbara M; Peck, Michael W

    2015-03-01

    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

  19. 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

  20. 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...

  1. 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

  2. Epsilon toxin: a fascinating pore-forming toxin.

    PubMed

    Popoff, Michel R

    2011-12-01

    Epsilon toxin (ETX) is produced by strains of Clostridium perfringens classified as type B or type D. ETX belongs to the heptameric ?-pore-forming toxins including aerolysin and Clostridium septicum alpha toxin, which are characterized by the formation of a pore through the plasma membrane of eukaryotic cells consisting in a ?-barrel of 14 amphipatic ? strands. By contrast to aerolysin and C. septicum alpha toxin, ETX is a much more potent toxin and is responsible for enterotoxemia in animals, mainly sheep. ETX induces perivascular edema in various tissues and accumulates in particular in the kidneys and brain, where it causes edema and necrotic lesions. ETX is able to pass through the blood-brain barrier and stimulate the release of glutamate, which accounts for the symptoms of nervous excitation observed in animal enterotoxemia. At the cellular level, ETX causes rapid swelling followed by cell death involving necrosis. The precise mode of action of ETX remains to be determined. ETX is a powerful toxin, however, it also represents a unique tool with which to vehicle drugs into the central nervous system or target glutamatergic neurons. PMID:21535407

  3. A functional recT gene for recombineering of Clostridium.

    PubMed

    Dong, Hongjun; Tao, Wenwen; Gong, Fuyu; Li, Yin; Zhang, Yanping

    2014-03-10

    Recombineering is an efficient genetic manipulation method employing the mechanism of phagenic RecT-mediated homologous recombination. To develop a recombineering method for Clostridium, a putative recT gene (CPF0939) from Clostridium perfringens genome was functionally verified in a clostridial host Clostridium acetobutylicum. We show that a short synthetic oligonucleotide can be introduced into the target site for specific point mutation. This functional recT gene would therefore contribute to development of recombineering tools for Clostridium. PMID:24384234

  4. Isolation of Nitrofurantoin-Resistant Mutants of Nitroreductase Producing Clostridium sp. Strains from the Human Intestinal Tract

    Microsoft Academic Search

    FATEMEH RAFII

    1998-01-01

    Five spontaneous nitrofurantoin-resistant mutants (one each of Clostridium leptum, Clostridium paraputrifi- cum, two other Clostridium spp. strains from the human intestinal microflora, and Clostridium perfringens ATCC 3626) were selected by growth on a nitrofurantoin-containing medium. All of the Clostridium wild-type and mutant strains produced nitroreductase, as was shown by the conversion of 4-nitrobenzoic acid to 4-aminobenzoic acid. High-performance liquid chromatography

  5. Clostridium bacteraemia characterised by 16S ribosomal RNA gene sequencing

    PubMed Central

    Woo, P C Y; Lau, S K P; Chan, K-m; Fung, A M Y; Tang, B S F; Yuen, K-y

    2005-01-01

    Background: Owing to problems in accurate species identification of the diverse genus clostridium, the epidemiology and pathogenicity of many species are not fully understood. Moreover, previous studies on clostridium bacteraemia have been limited and relied only on phenotypic species identification. Aims: To characterise the epidemiology, disease spectrum, and outcome of clostridium bacteraemia using 16S ribosomal RNA (rRNA) gene sequencing. Method: During a four year period (1998–2001), all cases of clostridium bacteraemia were prospectively studied and all “non-perfringensclostridium isolates identified to the species level by 16S rRNA gene sequencing. Results: Fifty one blood culture isolates were identified as Clostridium perfringens and 17 belonged to 11 other clostridium species. The first case of C disporicum infection and two cases of clostridium bacteraemia in children with intussusception were also described. Of the 68 clostridium isolates from 68 different patients, 38 were associated with clinically relevant bacteraemia. The gastrointestinal and hepatobiliary tracts were common sites of both underlying disease and portal of entry in these patients. Clostridium perfringens accounted for 79% of all clinically relevant bacteraemia, with the remainder caused by a diversity of species. The attributable mortality rate of clinically relevant clostridium bacteraemia was 29%. Younger age and underlying gastrointestinal/hepatobiliary tract disease were associated with mortality (p < 0.05). Conclusions: Patients with clinically relevant clostridium bacteraemia should be investigated for the presence of underlying disease processes in the gastrointestinal or hepatobiliary tracts. 16S rRNA gene analysis will continue to be useful in further understanding the pathogenicity of various clostridium species. PMID:15735165

  6. In vitro inhibition of growth of Escherichia coli, Salmonella Typhimurium, and Clostridia perfringens using Probiotics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella Typhimurium, Escherichia coli and Clostridium perfringens are pathogenic organisms found in horses [1] and they cause disease in animals or humans [2]. Due to concern over pathogens such as these, there is increasing interest in antimicrobial alternatives to prevent or reduce the prevalen...

  7. Monitoring of anti-C. perfringens bacteriophage CpV1 persistence in gastrointestinal tracts of broilers.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A factor limiting promotion of poultry products to the world market is any contamination of birds with pathogens, including Clostridium perfringens. The latter is often accountable for significant economical losses in production of commercial birds because of a possibility of the development of necr...

  8. Epsilon Aurigae

    NASA Astrophysics Data System (ADS)

    Turner, Rebecca; Price, A.; Henden, A.

    2009-05-01

    The IYA 2009 working group on Research Experiences for Students, Teachers, and Citizen-Scientists is planning a multi-year project involving the bright star Eps Aur. The project will go beyond simple observing and also include a major data analysis component. The goal is to introduce the participant to the full scientific process from background research to paper writing for a peer-reviewed journal. It begins with a 10 Star Training Program of several types of binary and transient variable stars that are easy to observe from suburban locations with the naked eye. Participants will be trained both in observing and also in basic data analysis of photometric datasets (light curve and period analysis). Eventually it will lead to a capstone project: monitoring the rare and mysterious 2009-2011 eclipse of Epsilon Aurigae. In the summer of IYA 2009, third-magnitude Eps Aur will experience its next eclipse, which occurs every 27.1 years and lasts 714 days, nearly two years. The star is bright enough to be seen with the naked eye from most urban areas. If fully funded, the project will also involve two public workshops on observing and data analysis in the summers of 2009 and 2010, respectively.

  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

    Shimamoto, S; Moriyama, R; Sugimoto, K; Miyata, S; Makino, S

    2001-06-01

    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 degrees 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 HgCl(2) 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. 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,

  11. 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,

  12. Classification of endospores of Bacillus and Clostridium species by FT-IR reflectance microspectroscopy and autoclaving

    Microsoft Academic Search

    D. L. Perkins; C. R. Lovell; B. V. Bronk; B. Setlow; P. Setlow; M. L. Myrick

    2005-01-01

    Fourier transform infrared reflectance microspectroscopy was applied to the classification of bacterial endospores, both untreated and those pretreated with autoclaving. Mid-infrared vibrational spectra were collected following minimal sample preparation from endospores of five species of bacteria: Bacillus cereus, Bacillus globigii, Bacillus megaterium, Bacillus subtilis, and Clostridium perfringens. The amide I and amide II regions were found to provide the best

  13. How to measure epsilon'/epsilon with lattice QCD

    SciTech Connect

    Sharpe, S.R.

    1987-04-01

    A pedagogical discussion is given of a lattice calculation of epsilon'. The method is outlined, and preliminary results are presented. They suggest that epsilon'/epsilon may be reduced from previous estimates by 60-70%.

  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. The Fractionation of Clostridium welchii e-Antigen on Cellulose Ion Exchangers

    Microsoft Academic Search

    R. O. Thomson

    1963-01-01

    SUMMARY A low-protein medium is described for the production of reasonable yields of Clostridium welchii (perfringens) e-prototoxin. The e-antigen in young cultures on this medium was virtually non-toxic, 99.Sy0 being in the form of prototoxin. Its purity, estimated in terms of the protein content of crystalline e-prototoxin, was 50 yo. Fractionation of concen- trates of the antigen on columns of

  17. Actin-specific ADP-ribosyltransferase produced by a Clostridium difficile strain.

    PubMed Central

    Popoff, M R; Rubin, E J; Gill, D M; Boquet, P

    1988-01-01

    By screening possible ADP-ribosyltransferase activities in culture supernatants from various Clostridium species, we have found one Clostridium difficile strain (CD196) (isolated in our laboratory) that is able to produce, in addition to toxins A and B, a new ADP-ribosyltransferase that was shown to covalently modify cell actin as Clostridium botulinum C2 or Clostridium perfringens E iota toxins do. The molecular weight of the CD196 ADP-ribosyltransferase (CDT) was determined to be 43 kilodaltons, and its isoelectric point was 7.8. No cytotoxic activity on Vero cells or lethal activity upon injection in mice was associated with this enzyme. CDT was neither related to C. difficile A or B toxins nor to C. botulinum C2 toxin component I. However, Vero cells cultivated in the presence of C. difficile B toxin had a lower amount of actin able to be ADP-ribosylated by CDT or C2 toxin in vitro. Antibodies raised against CDT reacted by immunoblot analysis with a 43-kilodalton protein of C. perfringens type E culture supernatant producing the iota toxin. Images PMID:3137166

  18. [Studies of necrotizing enteritis of suckling piglets (Cl. perfringens typc C enterotoxemia) in industrialized sow breeding units. 3. Experimental reproduction of the disease].

    PubMed

    Köhler, B; Rösch, B; Haase, H; Baumann, G

    1979-01-01

    Experimental reproduction of necrotising enteritis of sucking pigs was successfully achieved by using both Clostridium perfringens Type C strains, which had been isolated from sucking pigs with necrotising enteritis, and Type C strain 3628 of A.T.C.C. (sub-type C1). The lethal dose for sucking pigs was between 20 X 10(6) and 12 X 10(7) pathogens per animal. The disease could not even be induced by repeated application of no-bacterial toxin of Cl. perfringens Type C nor by administration of Cl. perfringens Type A strains which had been cultured from broilers with necrotising enteritis. Necrotising enteritis was found to develop in two phases in sucking pigs. First, the pathogen will deposit to the villous epithelium and then penetrate the superficial strata of the mocous membrane. In the second phase, the villous structure will be destroyed by the lethal, haemolysing, and necrotising toxins of Cl. perfringens. The role played by individual toxin fractions is discussed together with the importance of humoral and localised infection defence. Sucking pigs may be sufficiently protected against infection based on single or ten-fold lethal infectious dosage by two vaccinations of the mother animal, five and three weeks prior to parturition, using "Enterotoxaemia Vaccine Dessau bivalent". Infection then would not occur unless a hundredfold lethal dose was applied. Characteristics include diarrhoea, apathy, exhaustion, and death. PMID:228629

  19. Myonecrosis by Clostridium septicum in a dog, diagnosed by a new multiplex-PCR.

    PubMed

    Ribeiro, Márcio Garcia; Silva, Rodrigo Otávio Silveira; Pires, Prhiscylla Sadanã; Martinho, Anna Paula Vitirito; Lucas, Thays Mizuki; Teixeira, Ana Izabel Passarela; Paes, Antonio Carlos; Barros, Claudenice Batista; Lobato, Francisco Carlos Faria

    2012-10-01

    Clostridial myositis is an acute, generally fatal toxemia that is considered to be rare in pet animals. The present report describes an unusual canine clostridial myositis that was diagnosed by a new multiplex-PCR (mPCR) designed for simultaneous identification of Clostridium sordellii, Clostridium septicum, Clostridium perfringens type A, Clostridium chauvoei, and Clostridium novyi type A. A ten-month-old male Rottweiler dog, that had displayed lameness and swelling of the left limb for 12 h, was admitted to a veterinary hospital. The animal was weak, dyspneic and hyperthermic, and a clinical examination indicated the presence of gas and edema in the limb. Despite emergency treatment, the animal died in only a few minutes. Samples of muscular tissue from the necrotic area were aseptically collected and plated onto defibrinated sheep blood agar (5%) in anaerobic conditions. Colonies suggestive of Clostridium spp. were submitted to testing by multiplex-PCR. Impression smears of the tissues, visualized with Gram and also with panoptic stains, revealed long rod-shaped organisms, and specimens also tested positive using the fluorescent antibody technique (FAT). The FAT and mPCR tests enabled a diagnosis of C. septicum myonecrosis in the dog. PMID:22975141

  20. Gas discharge plasmas are effective in inactivating Bacillus and Clostridium spores.

    PubMed

    Tseng, Shawn; Abramzon, Nina; Jackson, James O; Lin, Wei-Jen

    2012-03-01

    Bacterial spores are the most resistant form of life and have been a major threat to public health and food safety. Nonthermal atmospheric gas discharge plasma is a novel sterilization method that leaves no chemical residue. In our study, a helium radio-frequency cold plasma jet was used to examine its sporicidal effect on selected strains of Bacillus and Clostridium. The species tested included Bacillus subtilis, Bacillus stearothermophilus, Clostridium sporogenes, Clostridium perfringens, Clostridium difficile, and Clostridium botulinum type A and type E. The plasmas were effective in inactivating selected Bacillus and Clostridia spores with D values (decimal reduction time) ranging from 2 to 8 min. Among all spores tested, C. botulinum type A and C. sporogenes were significantly more resistant to plasma inactivation than other species. Observations by phase contrast microscopy showed that B. subtilis spores were severely damaged by plasmas and the majority of the treated spores were unable to initiate the germination process. There was no detectable fragmentation of the DNA when the spores were treated for up to 20 min. The release of dipicolinic acid was observed almost immediately after the plasma treatment, indicating the spore envelope damage could occur quickly resulting in dipicolinic acid release and the reduction of spore resistance. PMID:22075631

  1. Clostridium botulinum type E occurs and grows in the alga Cladophora glomerata

    USGS Publications Warehouse

    Byappanahalli, M.N.; Whitman, R.L.

    2009-01-01

    In recent years, massive avian die-offs from Clostridium botulinum type E infection have occurred in the Sleeping Bear Dunes National Lakeshore (SLBE) area of Lake Michigan. These outbreaks have been coincidental with massive blooms of the green algae Cladophora, mostly Cladophora glomerata. We tested the hypothesis that Clostridium botulinum type E can grow under suitable conditions in these algal mats. In a lab mesocosm study, Cladophora from four outbreak-impacted beaches from SLBE were compared with four unimpacted beaches in the Milwaukee–Racine area for bontE gene of Clostridium botulinum. Frequency of the bontE gene was higher after incubation (25 °C for up to 6 weeks) of Cladophora from impacted vs. the unimpacted area. Since no type E gene was detected initially in Cladophora from any of the eight locations, we infer that the increased occurrence of type E gene arose from spore germination or vegetative Clostridium growth within the existing algal mats of SLBE. Moreover, we found that the congener Clostridium perfringens readily grows in mesocosms containing Cladophora.

  2. Quantitative real-time PCR assay for Clostridium septicum in poultry gangrenous dermatitis associated samples.

    PubMed

    Neumann, A P; Dunham, S M; Rehberger, T G; Siragusa, G R

    2010-08-01

    Clostridium septicum is a spore-forming anaerobe frequently implicated in cases of gangrenous dermatitis (GD) and other spontaneously occurring myonecrotic infections of poultry. Although C. septicum is readily cultured from diseased tissues it can be difficult to enumerate due to its tendency to swarm over the surface of agar plates. In this study a quantitative real-time PCR assay was developed in order to more accurately measure the levels of C. septicum in healthy as well as GD associated poultry samples. The assay was specifically designed to target the C. septicum alpha toxin gene, csa, which is, to our knowledge, carried by all strains of C. septicum and has been shown to be essential for virulence. Genomic DNAs from a diverse collection of bacterial species, including closely related Clostridium chauvoei, Clostridium carnis, Clostridium tertium as well as several strains of Clostridium perfringens, all failed to produce a positive reaction. An approximate reproducible limit of detection in spiked extracts of at least 10(3) cfu/g of C. septicum was observed for a variety of different sample types. C. septicum levels in broiler chicken field samples estimated from the results of qPCR were statistically correlated to culture based enumerations obtained from those same tissues. PMID:20399850

  3. Clostridium Difficile Infections

    MedlinePLUS

    Clostridium difficile (C. difficile) is a bacterium that causes diarrhea and more serious intestinal conditions such as colitis. Symptoms include Watery ... Nausea Abdominal pain or tenderness You might get C. difficile disease if you have an illness that ...

  4. Collagenase Clostridium Histolyticum Injection

    MedlinePLUS

    ... Clostridium histolyticum injection is also used to treat Peyronie's disease (a thickening of tissue [plaque] inside the ... the finger(s) to be straightened. In people with Peyronie's disease, it works by helping to break down ...

  5. Nosocomial Diarrhea: Evaluation and Treatment of Causes Other Than Clostridium difficile

    PubMed Central

    Polage, Christopher R.; Solnick, Jay V.; Cohen, Stuart H.

    2012-01-01

    Diarrhea is common among hospitalized patients but the causes are distinct from those of diarrhea in the community. We review existing data about the epidemiology of nosocomial diarrhea and summarize recent progress in understanding the mechanisms of diarrhea. Clinicians should recognize that most cases of nosocomial diarrhea have a noninfectious etiology, including medications, underlying illness, and enteral feeding. Apart from Clostridium difficile, the frequency of infectious causes such as norovirus and toxigenic strains of Clostridium perfringens, Klebsiella oxytoca, Staphylococcus aureus, and Bacteroides fragilis remains largely undefined and test availability is limited. Here we provide a practical approach to the evaluation and management of nosocomial diarrhea when tests for C. difficile are negative. PMID:22700831

  6. Structure of a C. perfringens enterotoxin mutant in complex with a modified Claudin-2 extracellular loop 2.

    PubMed

    Yelland, Tamas S; Naylor, Claire E; Bagoban, Tannya; Savva, Christos G; Moss, David S; McClane, Bruce A; Blasig, Ingolf E; Popoff, M; Basak, Ajit K

    2014-09-01

    CPE (Clostridium perfringens enterotoxin) is the major virulence determinant for C. perfringens type-A food poisoning, the second most common bacterial food-borne illness in the UK and USA. After binding to its receptors, which include particular human claudins, the toxin forms pores in the cell membrane. The mature pore apparently contains a hexamer of CPE, claudin and, possibly, occludin. The combination of high binding specificity with cytotoxicity has resulted in CPE being investigated, with some success, as a targeted cytotoxic agent for oncotherapy. In this paper, we present the X-ray crystallographic structure of CPE in complex with a peptide derived from extracellular loop 2 of a modified, CPE-binding Claudin-2, together with high-resolution native and pore-formation mutant structures. Our structure provides the first atomic-resolution data on any part of a claudin molecule and reveals that claudin's CPE-binding fingerprint (NPLVP) is in a tight turn conformation and binds, as expected, in CPE's C-terminal claudin-binding groove. The leucine and valine residues insert into the binding groove while the first residue, asparagine, tethers the peptide via an interaction with CPE's aspartate 225 and the two prolines are required to maintain the tight turn conformation. Understanding the structural basis of the contribution these residues make to binding will aid in engineering CPE to target tumor cells. PMID:25020226

  7. Diarylacylhydrazones: Clostridium-selective antibacterials with activity against stationary-phase cells.

    PubMed

    Chen, Chao; Dolla, Naveen K; Casadei, Gabriele; Bremner, John B; Lewis, Kim; Kelso, Michael J

    2014-01-15

    Current antibiotics for treating Clostridium difficile infections (CDI), that is, 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 Letter reports that certain diarylacylhydrazones identified during a high-throughput screening/counter-screening campaign show selective activity against two Clostridium species (C. difficile and Clostridium 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

  8. Congener specific polychlorinated biphenyl metabolism by human intestinal microbe Clostridium species: Comparison with human liver cell line-HepG2

    PubMed Central

    De, Supriyo; Ghosh, Somiranjan; Dutta, Sisir K.

    2015-01-01

    Polychlorobiphenyls (PCBs), which adversely affect human fetal and infant development, are endocrine disrupter and cause neurological disorders. They may also be carcinogenic. It is not known whether these effects are due to whole PCBs or to its metabolites, produced by the human gastrointestinal system primarily the liver and/or by intestinal microbes such as Clostridium sp. The available data show that Clostridium perfringens, the most prominent species of Clostridium occurs in the human gut. C. beijerinckii is a special type of Clostridium present in the gut of autistic children with late onset autism. Since mixed cultures are better PCB metabolizers than single cultures, mixed cultures of Clostridium were used in this work. The first step in PCB degradation is the removal of the chlorine atoms and then the breaking open of the phenyl ring leading to the final degradation product: CO2. In this study, GC-MS analyses were done to examine the effect of Clostridium sp. on PCB-153 and PCB-77 and the metabolites obtained with Clostridium sp. therein. In this paper, we report that the unlike human liver cells which cannot produce any PCB metabolites. Mixed Clostridium spp. can degrade these PCBs. Clostridium spp. and were able to dechlorinate PCB 153 (hexachlorobiphenyl) to pentachlorobiphenyl and PCB 77 (tetrachlorobiphenyl) to trichlorobiphenyl. Despite considerable absorption of PCB 153 (40%) and PCB 77 (50%) in 30 minutes and 1.5 hours respectively by human liver (HepG2) cells, they can not dechlorinate PCBs. It has been observed that slight differences in chemical structures of PCBs such as coplanar (PCB-77) vs. non-coplanar (PCB-153) has significant metabolic effects.

  9. Epsilon Aurigae. [eclipsing binary system

    NASA Technical Reports Server (NTRS)

    Chapman, R. D.

    1985-01-01

    In April 1984, fourth contact ended the two year long eclipse of Epsilon Aurigae. An astrometric study of the study of the system was carried out by Van de kamp (1978) leading to the conclusion that the orbit is seen very close to edge on. The eclipse was monitored by a number of groups from the ground and from spacecraft such as the IUE. Ultraviolet observations of the system from IUE have thrown new light on the nature of the system that led to the conclusion that the secondary object is probably a cold, dusty accretion disk surrounding a star that is completely hidden inside the disk.

  10. Clostridium difficile infection

    PubMed Central

    Viswanathan, VK; Mallozzi, MJ

    2010-01-01

    Clostridium difficile infection (CDI) is the primary cause of antibiotic-associated diarrhea and is a significant nosocomial disease. In the past ten years, variant toxin-producing strains of C. difficile have emerged, that have been associated with severe disease as well as outbreaks worldwide. This review summarizes current information on C. difficile pathogenesis and disease, and highlights interventions used to combat single and recurrent episodes of CDI. PMID:21327030

  11. 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

  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

    and D Clostr1dium ~fi t 1 . The eases t f fC. ~fi est neutralizing ant1bod1es to foals via the colostrum of vaccinated mares was also evaluated. The 18 mares used in this study were in a herd used for teaching purposes by the College of Veterinary... from mares within 24 hours post partum when possible. All mare and foal sera and colostrum samples were assayed for toxin neutral1zing antibodies by mouse inoculation. The horses involved in this study produced ant1body responses to vaccination ith...

  13. Enteritis associated with Clostridium perfringens type A in 9-month-old calves

    PubMed Central

    Savic, Bozidar; Prodanovic, Radisa; Ivetic, Vojin; Radanovic, Oliver; Bojkovski, Jovan

    2012-01-01

    Four 9-month-old Simmental male calves were presented with a history of sudden death. The necropsy and microscopic findings allowed a diagnosis of enteritis and severe intraluminal hemorrhage with blood clots in the jejunum, suggestive of jejunal hemorrhage syndrome. PMID:22851779

  14. Predictive model for growth of Clostridium perfringens during cooling of cooked ground chicken

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional methodologies for development of microbial growth models under dynamic temperature conditions do not take into account the organism’s prior history. Such models were shown to be inadequate in predicting growth of the organisms under dynamic conditions commonly encountered in the food ind...

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

    E-print Network

    Chen, Chang-Hsin 1977-

    2012-07-27

    immunogen to APCs through the CD40 pathway will improve protection against NE in chickens. One agonistic monoclonal anti-chCD40 antibody (designated 2C5) was produced and characterized. 2C5 not only detected expression of chCD40 on chicken APCs, but also...

  16. 9 CFR 113.111 - Clostridium Perfringens Type C Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 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 each serial...

  17. 9 CFR 113.111 - Clostridium Perfringens Type C Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 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 each serial...

  18. 9 CFR 113.112 - Clostridium Perfringens Type D Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 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 each serial...

  19. 9 CFR 113.111 - Clostridium Perfringens Type C Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 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 each serial...

  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. PREDICTIVE MODEL FOR GROWTH OF CLOSTRIDIUM PERFRINGENS DURING COOLING OF COOKED GROUND CHICKEN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional methodologies for development of microbial growth models under dynamic temperature conditions do not take adequate account for the organism’s history. Such models were shown to be inadequate in predicting growth of the organisms under dynamic conditions commonly encountered in the food i...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Scrapple is an ethnic food produced/consumed almost exclusively in the Middle Atlantic states of the U.S. It is typically made from ground pork trimmings, seasonings, cornmeal, and flour. This mixture is cooked and then shaped into loaves that are cooled and subsequently stored refrigerated until sl...

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

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...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...

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

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...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...

  5. 9 CFR 113.112 - Clostridium Perfringens Type D Toxoid and Bacterin-Toxoid.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...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...

  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. Comparative in vitro activities of LFF571 against Clostridium difficile and 630 other intestinal strains of aerobic and anaerobic bacteria.

    PubMed

    Citron, Diane M; Tyrrell, Kerin L; Merriam, C Vreni; Goldstein, Ellie J C

    2012-05-01

    The in vitro activities of LFF571, a novel analog of GE2270A that inhibits bacterial growth by binding with high affinity for protein synthesis elongation factor Tu, fidaxomicin, and 10 other antimicrobial agents were determined against 50 strains of Clostridium difficile and 630 other anaerobic and aerobic organisms of intestinal origin. LFF571 possesses potent activity against C. difficile and most other Gram-positive anaerobes (MIC(90), ? 0.25 ?g/ml), with the exception of bifidobacteria and lactobacilli. The MIC(90)s for aerobes, including enterococci, Staphylococcus aureus (as well as methicillin-resistant S. aureus [MRSA] isolates), Streptococcus pyogenes, and other streptococci were 0.06, 0.125, 2, and 8 ?g/ml, respectively. Comparatively, fidaxomicin showed variable activity against Gram-positive organisms: MIC(90)s against C. difficile, Clostridium perfringens, and Bifidobacterium spp. were 0.5, ? 0.015, and 0.125 ?g/ml, respectively, but >32 ?g/ml against Clostridium ramosum and Clostridium innocuum. MIC(90) for S. pyogenes and other streptococci was 16 and >32 ?g/ml, respectively. LFF571 and fidaxomicin were generally less active against Gram-negative anaerobes. PMID:22290948

  9. Bifidobacterium, Bacteroides, and Clostridium spp. in fecal samples from breast-fed and bottle-fed infants with and without iron supplement.

    PubMed Central

    Mevissen-Verhage, E A; Marcelis, J H; de Vos, M N; Harmsen-van Amerongen, W C; Verhoef, J

    1987-01-01

    Bifidobacterium, Bacteroides, and Clostridium spp. isolated from the feces of 23 neonates during the first 3 months of life were identified. Of the 23 neonates, 10 were breast fed, 6 received an infant formula with iron supplement (5 mg/liter), and 7 received the formula without iron supplement (iron concentration, less than 0.5 mg/liter). The Bifidobacterium spp. most frequently isolated from the three groups of infants were B. longum, B. breve, B. adolescentis, and B. bifidum. The bacteroides spp. most frequently isolated were B. fragilis and B. vulgatus. The most common Clostridium sp. in the three groups of infants was C. perfringens. The type of milk did not select for species of Bifidobacterium, Bacteroides, or Clostridium, except for Clostridium butyricum, which was isolated significantly more often from bottle-fed infants with iron supplement than from the other groups, and Clostridium tertium, which was more often isolated from breast-fed infants. The species of the three anaerobic genera did not persist for a long period of time in the three groups of infants. PMID:3818925

  10. Epsilon Aurigae Eclipse 2009 - Ingress

    NASA Astrophysics Data System (ADS)

    Hopkins, Jeffrey L.; Stencel, Robert E.; Leadbeater, Robin; Beckmann, Paul J.; Buil, Christian; Collins, Donald; Colombo, Tiziano; Garrel, Thierry; Gorodenski, Stanley; Gudmundsson, Snaevarr; Karlsson, Mukund Kurtadikar; Lindberg, Hans-Goran; Loughney, Des; Mauclaire, Benji; McCandless, Brian E.; Melillo, Frank J.; Miles, Richard; Pearson, Robert T.; Samolyk, Gerard; Schanne, Lothar; Strikis, Iakovos Marios; Teyssier, François; Thizy, Olivier

    The mysterious star system epsilon Aurigae undergoes an eclipse every 27.1 years that lasts nearly two years. The most recent eclipse started during the late summer of 2009. An international campaign for observing this eclipse was created in 2006, with a web site for information and, to-date, 17 periodic newsletters for details, as well as a Yahoo forum List for immediate announcements and comments. Photometric data in the UBVRIJH bands have been submitted. Ingress occurred with first contact in the V band estimated at the second week of 2009 August and second contact estimated at 2010 mid-January. Spectroscopic data were also obtained during ingress. Spectroscopic data have been provided in the potassium I region, hydrogen alpha and beta regions and sodium D line region of the star system's spectrum. In this paper we describe details of observations and preliminary analysis during ingress and second contact. We introduce the observers and discuss plans for observing throughout totality and the end of the eclipse in 2011.

  11. The search for companions to Epsilon Eridani.

    PubMed

    Lawton, A T; Wright, P

    1990-12-01

    The authors review efforts to examine the star Epsilon Eridani and determine the possibility for the existence of an Earth-like planet. Early data indicated that there must be a habitable ecosphere about 82.5 million Km from the primary. Research into the existence of another planetary system determined that Epsilon Eridani was a binary star with an Oort cloud system, indicating the possibility of planet formation. A review of the evidence suggests that the presence of the small red Dwarf companion star precludes the existence of a planetary system surrounding Epsilon Eridani. It is suggested that observations continue to provide further data about the formation of binary systems. PMID:11540498

  12. Antibiotic-Associated Diarrhea: Candidate Organisms other than Clostridium Difficile

    PubMed Central

    Song, Hyun Joo; Jung, Sung-Ae; Choi, Hee Jung; Lee, Mi Ae; Ryu, Kum Hei; Kim, Seong-Eun; Yoo, Kwon

    2008-01-01

    Backgraound/Aims The direct toxic effects of antibiotics on the intestine can alter digestive functions and cause pathogenic bacterial overgrowth leading to antibiotic-associated diarrhea (AAD). Clostridium Difficile (C. Difficile) is widely known to be responsible for 10~20% of AAD cases. However, Klebsiella oxytoca, Clostridium perfringens, Staphylococcus aureus, and Candida species might also contribute to AAD. Methods We prospectively analyzed the organisms in stool and colon tissue cultures with a C. Difficile toxin A assay in patients with AAD between May and December 2005. In addition, we performed the C. Difficile toxin A assays using an enzyme-linked fluorescent assay technique. Patients were enrolled who had diarrhea with more than three stools per day for at least 2 days after the initiation of antibiotic treatment for up to 6~8 weeks after antibiotic discontinuation. Results Among 38 patients (mean age 59±18 years, M:F=18:20), the organism isolation rates were 28.9% (11/38) for stool culture, 18.4% (7/38) for colon tissue cultures and 13.2% (5/38) for the C. Difficile toxin A assay. The overall rate of identification of organisms was 50.0% (19/38). Of the five patients that had a positive result by the C. Difficile toxin A assay, two had no organism isolated by the stool or colon tissue culture. The organisms isolated from the stool cultures were C. Difficile (4), Klebsiella pneumoniae (K. pneumoniae) (3), Candida species (3), and Staphylococcus aureus (1). C. Difficile (4) and K. pneumoniae (3) were isolated from the colon tissue culture. Conclusions For C. Difficile negative AAD patients, K. pneumoniae, Candida species, and Staphylococcus aureus were found to be potential causative organisms. PMID:18363274

  13. Vaccination of turkeys with Clostridium septicum bacterin-toxoid: evaluation of protection against clostridial dermatitis.

    PubMed

    Thachil, Anil J; McComb, Brian; Kromm, Michelle; Nagaraja, Kakambi V

    2013-06-01

    Clostridial dermatitis is an acute disease causing high mortality in turkeys. Both Clostridium septicum and Clostridium pefringens have been isolated from these cases; however, reports from several diagnostic laboratories indicate an increased isolation rate of C septicum compared with C. perfringens from cases of clostridial dermatitis in recent years. Previous studies suggested C. septicum was more potent than C. perfringens in causing clostridial dermatitis in turkeys. The objective of this study was to develop and evaluate the use of a C. septicum bacterin-toxoid to control clostridial dermatitis in turkeys. A C. septicum bacterin-toxoid was prepared and was initially tested in 6-wk-old commercial turkeys under laboratory conditions for its safety and efficacy. Subsequently, the bacterin-toxoid was evaluated for use in commercial turkey farms with a consistent history of clostridial dermatitis. Birds in the field were vaccinated subcutaneously once at 6 wk of age with C. septicum bacterin-toxoid, and then mortality in both vaccinated and unvaccinated groups was recorded and compared. Blood samples from birds in both groups were examined using ELISA to detect antibody response to the C. septicum toxoid. The C. septicum bacterin-toxoid was found to be safe and to elicit antibodies against the toxoid. In vaccinated commercial turkeys, control of clostridial dermatitis was achieved via antibiotic use and clostridial dermatitis mortality was significantly reduced compared with that of birds in the unvaccinated group. The C. septicum bacterin-toxoid seems to be a valuable tool for the turkey industry to reduce losses due to clostridial dermatitis. PMID:24689176

  14. JAMA Patient Page: Clostridium Difficile Colitis

    MedlinePLUS

    ... of the American Medical Association JAMA PATIENT PAGE Clostridium difficile Colitis C olitis (inflammation of the colon) can ... as a result of infection with the bacterium Clostridium difficile (also known as C difficile and C diff ). ...

  15. Faltings plus epsilon, Wiles plus epsilon, and the Generalized Fermat Equation

    E-print Network

    Darmon, Henri

    Faltings plus epsilon, Wiles plus epsilon, and the Generalized Fermat Equation H. Darmon September 9, 2007 Wiles' proof of Fermat's Last Theorem puts to rest one of the most famous unsolved problems celebrating Wiles' achieve- ment, one also feels a twinge of regret at Fermat's demise. Is the Holy Grail

  16. Antigenic Formulæ for the Genus Clostridium

    Microsoft Academic Search

    Raouf S. Moussa

    1958-01-01

    ANTIGENIC formulæ for Clostridium sporogenes, Clostridium histolyticum and Clostridium parabotulinum were devised by Mandia and Bruner1 and by Mandia2-4. These were restricted to the heat-labile antigens and excluded the O- and the spore- (S-) antigens.

  17. 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

  18. Isolation of Clostridium thermocellum auxotrophs

    SciTech Connect

    Mendez, B.S.; Gomez, R.F.

    1982-02-01

    The conversion of biomass of fuels and chemical feedstocks by microbial fermentation offers the potential of solving two of today's important problems: waste accumulation and exhaustion of fossil fuels. Microorganisms with the capabilities of converting biomass components such as cellulos and hemicellulose to chemicals and fuels in a single step are of particular interest. One such microorganism is Clostridium thermocellum, a thermophilic anaerobe which degrades cellulose to ethanol and organic acids. For efficient industrial use, the cellulolytic capacity of this strain must be improved by genetic means. Spontaneous and UV irradiation-induced auxotrophic mutants of Clostridium thermocellum, an anaerobic cellulolytic thermophile, were isolated after penicillin enrichment in a chemically defined medium.

  19. Epsilon substitution method for elementary analysis

    Microsoft Academic Search

    Grigori Mints; Sergei Tupailo; Wilfried Buchholz

    1996-01-01

    We formulate epsilon substitution method for elementary analysisEA (second order arithmetic with comprehension for arithmetical formulas with predicate parameters). Two proofs of its termination are presented. One uses embedding into ramified system of level one and cutelimination for this system. The second proof uses non-effective continuity argument.

  20. Epsilon-insensitive fuzzy c-regression models: introduction to epsilon-insensitive fuzzy modeling.

    PubMed

    Leski, Jacek M

    2004-02-01

    This paper introduces a new epsilon-insensitive fuzzy c-regression models (epsilonFCRM), that can be used in fuzzy modeling. To fit these regression models to real data, a weighted epsilon-insensitive loss function is used. The proposed method make it possible to exclude an intrinsic inconsistency of fuzzy modeling, where crisp loss function (usually quadratic) is used to match real data and the fuzzy model. The epsilon-insensitive fuzzy modeling is based on human thinking and learning. This method allows easy control of generalization ability and outliers robustness. This approach leads to c simultaneous quadratic programming problems with bound constraints and one linear equality constraint. To solve this problem, computationally efficient numerical method, called incremental learning, is proposed. Finally, examples are given to demonstrate the validity of introduced approach to fuzzy modeling. PMID:15369046

  1. MLST analysis reveals a highly conserved core genome among poultry isolates of Clostridium septicum.

    PubMed

    Neumann, Anthony P; Rehberger, Thomas G

    2009-06-01

    Clostridium septicum is a highly virulent, anaerobic bacterium capable of establishing necrotizing tissue infections and forming heat resistant endospores. Disease is primarily facilitated by secretion of numerous toxic products including a lethal pore-forming cytolysin. Spontaneously occurring clostridial myonecrosis involving C. septicum has recently reemerged as a concern for many poultry producers. However, despite its increasing prevalence, the epidemiology of infection and population structure of C. septicum remains largely unknown. In this study a multilocus sequence typing (MLST) approach was utilized to examine evolutionary relationships within a diverse collection of C. septicum isolates recovered from poultry flocks experiencing episodes of gangrenous dermatitis. The 109 isolates examined represented 42 turkey flocks and 24 different flocks of broiler chickens as well as C. septicum type strain, ATCC 12464. Isolates were recovered predominantly from gangrenous lesions although isolates from livers, gastrointestinal tracts, spleens and blood were included. The loci analyzed were csa, the major lethal toxin produced by C. septicum, and the housekeeping genes gyrA, groEL, dnaK, recA, tpi, ddl, colA and glpK. These loci were included in part because of their previous use in MLST analysis of Clostridium perfringens and Clostridium difficile. Results indicated a high level of conservation present within these housekeeping gene fragments when compared to what has been previously reported for the aforementioned clostridia. Of the 5352 bp of sequence data examined for each isolate, 99.7% (5335/5352) was absolutely conserved among the 109 isolates. Only one of the ten unique sequence types, or allelic profiles, identified among the isolates was recovered from both turkeys and broiler chickens suggesting some host species preference. Phylogenetic analyses identified two unique clusters, or clonal complexes, among these poultry isolates which may have important epidemiological implications for poultry producers in the United States. This work indicates a predominantly clonal population structure for C. septicum although some evidence of recombination was also observed. PMID:19402197

  2. Survey of neuraminidase production by Clostridium butyricum, Clostridium beijerinckii, and Clostridium difficile strains from clinical and nonclinical sources.

    PubMed Central

    Popoff, M R; Dodin, A

    1985-01-01

    Neuraminidase production was investigated in 57 Clostridium butyricum strains, 16 Clostridium beijerinckii strains, and 25 Clostridium difficile strains. Neuraminidase activity was found only in C. butyricum strains originating from one human newborn with neonatal necrotizing enterocolitis, two newborns with hemorrhagic colitis, one infected placenta, and one adult with peritonitis, It was concluded that neuraminidase was not a major virulence factor in C. butyricum strains. PMID:4056013

  3. Establishment of hybridomas secreting monoclonal antibodies against C epsilon 2 and C epsilon 4 domains of human IgE.

    PubMed

    Ichimori, Y; Kurokawa, T; Ikeyama, S; Sasada, R; Tsukamoto, K

    1985-01-01

    Three kinds of hybridomas secreting monoclonal antibodies (MAbs) against the epsilon chain of human IgE were constructed by somatic cell hybridization between mouse myeloma P3U1 cells and spleen cells from BALB/c mice immunized with human IgE purified from the culture supernatant of U-266 cells. These MAbs were used effectively for the purification and determination of human IgE. The recognition site in the IgE molecule of each antibody was examined by using various epsilon chain fragment peptides produced in Escherichia coli. From these experiments, it was suggested that one recognized C epsilon 2 and the second C epsilon 4. The third did not recognize the C epsilon 1-C epsilon 4 domains of the recombinant epsilon chain from E. coli, although it bound to the epsilon chain of natural human IgE. PMID:2578427

  4. A DIM CANDIDATE COMPANION TO {epsilon} CEPHEI

    SciTech Connect

    Mawet, D. [European Southern Observatory, Alonso de Cordova 3107, Vitacura, Santiago (Chile); Mennesson, B.; Serabyn, E.; Stapelfeldt, K. [Jet Propulsion Laboratory, California Institute of Technology, 4800 Oak Grove Drive, Pasadena, CA 91109 (United States); Absil, O. [Institut d'Astrophysique et de Geophysique de Liege, University of Liege, 17 Allee du 6 Aout, 4000 Sart Tilman (Belgium)

    2011-09-01

    Using a vector vortex coronagraph behind the 1.5 m well-corrected subaperture (WCS) at Palomar, we detected a second object very close to {epsilon} Cephei, a {delta} Scuti F0 IV star. The candidate companion, {approx}50 times fainter than {epsilon} Cephei, if physically associated, is a late-type K or early M star, and lies at an angular separation of 330 mas, or 1.1 {lambda}/D for the WCS, making it the smallest angle detection ever realized with a coronagraph in terms of {lambda}/D units. The projected separation of the putative companion is {approx}8.6 AU, most likely on a highly eccentric orbit. The recently detected near-infrared excess is thus likely not due to hot dust. Moreover, we also show that the previously reported IRAS 60 {mu}m excess was due to source confusion on the galactic plane.

  5. Recent Spectroscopy of Epsilon Aurigae in Eclipse

    NASA Astrophysics Data System (ADS)

    Ketzeback, William F.

    2011-01-01

    Modern digital spectroscopy of bright stars not only opens opportunities in asteroseismic and planet detection areas, but an eclipsing binary like epsilon Aurigae provides a CATscan-like dissection of the dark disk now transiting the super-large F star. I will report on optical and near-infrared spectroscopic monitoring being conducted at the ARC 4 meter at Apache Point Observatory and report on major changes in the spectra due to eclipse, assigning changes to the disk insofar as possible.

  6. Clostridium difficile-associated colitis.

    PubMed Central

    Hull, Mark W.; Beck, Paul L.

    2004-01-01

    OBJECTIVE: To review the basic microbiology, pathogenesis of disease, and diagnosis of the nosocomial pathogen Clostridium difficile and to examine therapies recommended by the Canadian Task Force on Preventive Health Care. QUALITY OF EVIDENCE MEDLINE: was searched using MeSH headings. Controlled trials for therapy were sought, but case-control studies and observational reviews were included. MAIN MESSAGE: Clostridium difficile causes approximately 20% of cases of diarrhea associated with antibiotics, including clindamycin and the second- and third-generation cephalosporins. Diarrhea is usually mild, but can be severe; extreme cases develop toxic megacolon. Diagnosis is dependent on demonstrating presence of clostridial toxin in stool specimens or of pseudomembranes through sigmoidoscopy. First-line therapy for C. difficile diarrhea is restricted to metronidazole. Second-line therapy for treatment failure is vancomycin. For relapse, a second course of metronidazole is recommended; tapering courses of vancomycin and probiotics are used for multiple recurrences. CONCLUSION: Clostridium difficile is an important nosocomial pathogen requiring prudent use of antibiotics and strict infection-control policies to prevent large health care costs. PMID:15597970

  7. Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration

    PubMed Central

    Al-Hinai, Mohab A.; Fast, Alan G.

    2012-01-01

    Isolation of Clostridium mutants based on gene replacement via allelic exchange remains a major limitation for this important genus. Use of a heterologous counterselection marker can facilitate the identification of the generally rare allelic exchange events. We report on the development of an inducible counterselection marker and describe its utility and broad potential in quickly and efficiently generating markerless DNA deletions and integrations at any genomic locus without the need for auxotrophic mutants or the use of the mobile group II introns. This system is based on a codon-optimized mazF toxin gene from Escherichia coli under the control of a lactose-inducible promoter from Clostridium perfringens. This system is potentially applicable to almost all members of the genus Clostridium due to their similarly low genomic GC content and comparable codon usage. We isolated all allelic-exchange-based gene deletions (ca_p0167, sigF, and sigK) or disruptions (ca_p0157 and sigF) we attempted and integrated a 3.6-kb heterologous DNA sequence (made up of a Clostridium ljungdahlii 2.1-kb formate dehydrogenase [fdh] gene plus a FLP recombination target [FRT]-flanked thiamphenicol resistance marker) into the Clostridium acetobutylicum chromosome. Furthermore, we report on the development of a plasmid system with inducible segregational instability, thus enabling efficient deployment of the FLP-FRT system to generate markerless deletion or integration mutants. This enabled expeditious deletion of the thiamphenicol resistance marker from the fdh integrant strain as well as the sigK deletion strain. More generally, our system can potentially be applied to other organisms with underdeveloped genetic tools. PMID:22983967

  8. Clostridium difficile causing acute renal failure: Case presentation and review

    Microsoft Academic Search

    Jasmin Arrich; Gottfried H. Sodeck; Gürkan Sengölge; Christoforos Konnaris; Marcus Müllner; Anton N. Laggner; Hans Domanovits

    Abstract Abstract Abstract Abstract AIM: Clostridium difficile infection is primarily a nosocomial infection but asymptomatic carriers of Clostridium difficile can be found in up to 5% of the general population. Ampicillin, cephalosporins and clindamycin are the antibiotics that are most frequently associated with Clostridium difficile-associated diarrhea or colitis. Little is known about acute renal failure as a consequence of Clostridium

  9. Successful management of spontaneous Clostridium septicum myonecrosis.

    PubMed

    Chipp, E; Phillips, C; Rubin, P

    2009-10-01

    Spontaneous Clostridium septicum myonecrosis, or gas gangrene, is an extremely rare soft tissue infection associated with malignancy and immunosuppression. Even with appropriate treatment the mortality rate approaches 100%. We present the case of a patient who presented with fulminant Clostridium septicum sepsis and myonecrosis who was successfully treated and made a full recovery within two weeks of the initial episode. PMID:18555761

  10. Fatal neutropenic enterocolitis due to clostridium septicum.

    PubMed

    Shah, B K; KC, R

    2011-10-01

    We describe a case of Clostridium septicum enterocolitis in a patient with pre-B acute lymphoblastic leukaemia undergoing autologous stem cell transplant. In the setting of neutropenia, Clostridium septicum should be suspected in patients who develop signs and symptoms of acute abdomen. PMID:22519241

  11. Epsilon Aur monitoring during predicted pulsation phase

    NASA Astrophysics Data System (ADS)

    Waagen, Elizabeth O.; Templeton, Matthew R.

    2014-09-01

    Dr. Robert Stencel (University of Denver Astronomy Program) has requested that AAVSO observers monitor epsilon Aurigae from now through the end of the observing season. "Studies of the long-term, out-of-eclipse photometry of this enigmatic binary suggest that intervals of coherent pulsation occur at roughly 1/3 of the 27.1-year orbital period. Kloppenborg, et al. noted that stable variation patterns develop at 3,200-day intervals' implying that 'the next span of dates when such events might happen are circa JD ~2457000 (2014 December)'. "These out-of-eclipse light variations often have amplitudes of ~0.1 magnitude in U, and ~0.05 in V, with characteristic timescales of 60-100 days. The AAVSO light curve data to the present may indicate that this coherent phenomenon has begun, but we encourage renewed efforts by observers...to help deduce whether these events are internal to the F star, or externally-driven by tidal interaction with the companion star." Nightly observations or one observation every few days (CCD/PEP/DSLR, VUBR (amplitude too small for visual)) are requested. Finder charts with sequence may be created using the AAVSO Variable Star Plotter (http://www.aavso.org/vsp). Observations should be submitted to the AAVSO International Database. Epsilon Aur was the subject of major international campaigns and the AAVSO's Citizen Sky project as it went through its 27.1-year eclipse in 2009-2011. Over 700 observers worldwide submitted over 20,000 multicolor observations to the AAVSO International Database for this project. Much information on eps Aur is available from the AAVSO, including material on the Citizen Sky website (http://www.aavso.org/epsilon-aurigae and http://www.citizensky.org/content/star-our-project). The Journal of the AAVSO, Volume 40, No. 2 (2012) was devoted to discussion of and research results from this event. See full Alert Notice for more details and observations.

  12. 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.

  13. 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.

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

    SciTech Connect

    Worcester, E.T.

    2009-10-01

    The authors 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 Fermi National Accelerator Laboratory during 1996, 1997, and 1999. This dataset contains about 15 million K {yields} {pi}{sup 0}{pi}{sup 0} and 70 million K {yields} {pi}{sup +}{pi}{sup -} decays. They measure the direct CP violation parameter Re({epsilon}'/{epsilon}) = (19.2 {+-} 2.1) x 10{sup -4}. they find the K{sub L}-K{sub S} mass difference {Delta}m = (5265 {+-} 10) x 10{sup 6} {bar h}s{sup -1} and the K{sub S} lifetime {tau}{sub S} = (89.62 {+-} 0.05) x 10{sup -12} s. They 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. Classical closure theory and Lam's interpretation of epsilon-RNG

    NASA Technical Reports Server (NTRS)

    Zhou, YE

    1995-01-01

    Lam's phenomenological epsilon-renormalization group (RNG) model is quite different from the other members of that group. It does not make use of the correspondence principle and the epsilon-expansion procedure. We demonstrate that Lam's epsilon-RNG model is essentially the physical space version of the classical closure theory in spectral space and consider the corresponding treatment of the eddy viscosity and energy backscatter.

  16. Epsilon Metal Summary Report FY 2011

    SciTech Connect

    Strachan, Denis M.; Crum, Jarrod V.; Zumhoff, Mac R.; Bovaird, Chase C.; Windisch, Charles F.; Riley, Brian J.

    2011-09-30

    The Epsilon-metal ({var_epsilon}-metal) phase was selected in FY 2009 as a potential waste form to for immobilizing the noble metals found in the undissolved solids + aqueous stream, and the soluble Tc from ion-exchange process, each resulting from proposed aqueous reprocessing. {var_epsilon}-metal phase is observed in used nuclear fuel and the natural reactors of Oklobono in Gabon, where the long-term corrosion behavior was demonstrated. This makes {var_epsilon}-metal a very attractive waste form. Last fiscal year, {var_epsilon}-metal was successfully fabricated by combining the five-metals, Mo, Ru, Rh, Pd and Re (surrogate for Tc), into pellets followed by consolidation with an arc melter. The arc melter produced fully dense samples with the epsilon structure. However, some chemistry differences were observed in the microstructure that resulted in regions rich in Re and Mo, and others rich in Pd, while Ru and Rh remained fairly constant throughout. This year, thermal stability (air), and corrosion testing of the samples fabricated by arc melting were the main focus for experimental work. Thermal stability was measured with a differential scanning calorimeter - thermogravimetric analyzer, by both ramp heating as well as step heating. There is clear evidence during the ramp heating experiment of an exothermic event + a weight loss peak both beginning at {approx}700 C. Step heating showed an oxidation event at {approx}690 C with minimal weight gain that occurs just before the weight loss event at 700 C. The conclusion being that the e-metal begins to oxidize and then become volatile. These findings are useful for considering the effects of voloxidation process. Three different pellets were subjected to electrochemical testing to study the corrosion behavior of the epsilon-metal phase in various conditions, namely acidic, basic, saline, and inert. Test was done according to an interim procedure developed for the alloy metal waste form. First an open circuit potential was measured, followed by linear polarization sweeps. The linear polarization sweep range was the Tafel equation was fit to the linear polarization sweep data to determine the corrosion rate of each pellet in each test solution. The average calculated corrosion rates of the three pellets according to solution conditions were: -1.91 x 10{sup -4} mm/yr (0.001 M NaOH), -1.48 x 10{sup -3} mm/yr (0.01 M NaCl), -8.77 x 10{sup -4} mm/yr (0.001 M H{sub 2}SO{sub 4}), -2.09 x 10{sup -3} mm/yr (0.001 M NaOH + 0.01 M NaCl), and -1.54 x 10{sup -3} mm/yr (0.001 M H{sub 2}SO{sub 4} + 0.01 M NaCl). Three single-pass flow through (SPFT) test were conducted at a flow rate of 10 ml/day, at 90 C, and pH of 2.5, 7.0, and 9.0 for up to 322 days. Results of the tests indicate that dissolution rates were 5 x 10{sup -4} g m{sup 2} d{sup -1} at pH 9.0, 1.2 x 10{sup -4} g m{sup -2} d{sup -1} at pH 7.0, and 2 x 10{sup -4} g m{sup -2} d{sup -1} at pH 2.5. The sample used for the pH 7.0 SPFT test contains extra Re compared to samples used for the other two SPFT test, which came from a single pellet. The corrosion data measured this year indicate that the {var_epsilon}-metal phase is chemically durable. The two chemically different phases, but structurally the same, behave differently during dissolution according to the microstructure changes observed in both the electrochemical and in SPFT test. Characterization of the test specimens after testing suggests that the dissolution is complex and involves oxidative dissolution followed by precipitation of both oxide and metallic phases. These data suggest that the dissolution in the electrochemical and SPFT tests is different; a process that needs further investigation.

  17. 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 common methodology for developing growth models for dynamic environments is to first determine growth kinetic parameters from a series of growth experiments conducted within well-defined environments under isothermal conditions. Subsequently, secondary models are developed to evaluate the effec...

  18. Biosynthesis of daunorubicin glycosides: role of epsilon-rhodomycinone.

    PubMed Central

    McGuire, J C; Thomas, M C; Stroshane, R M; Hamilton, B K; White, R J

    1980-01-01

    Daunorubicin (daunomycin; NSC 82151) is a fermentation-derived anthracycline antibiotic that is clinically useful in the treatment of human leukemias. Daunorubicin itself is found rarely in microbial fermentations, but is present normally in the form of glycoside derivatives that yield the free drug on simple acid hydrolysis. A major by-product of daunorubicin fermentations is usually the structurally related anthracyclinone epsilon-rhodomycinone. We have used mutants of a daunorubicin-producing Streptomyces species to study the biosynthetic relationship between epsilon-rhodomycinone and daunorubicin. We found that exogenously added epsilon-rhodomycinone can be converted to daunorubicin glycosides by a nonproducing mutant and by a mutant that produces daunorubicin glycosides but not epsilon-rhoeomycinone. Molar conversion efficiences were in the 15 to 30% range. The latter mutant was also shown to convert exogenous 14C-labeled epsilon-rhodomycinone to 14C-labeled daunorubicin glycosides, again at conversion efficiencies of about 25%. The same biotransformation was observed with daunorubicin production strain C5, which normally accumulates both epsilon-rhodomycinone and daunorubicin glycosides. A significant percentage (16 to 37%) of exogenously added epsilon-[14C]rhodomycinone was metabolized by strain C5, and 22 to 32% of the metabolized radioactivity could be recovered as daunorubicin glycosides. A mathematical model of epsilon-rhodomycinone metabolism was constructed based on plausible assumptions concerning the kinetics of epsilon-rhodomycinone accumulation and catabolsim. When analyzed according to this model, our data indicate that most (63 to 73%), but not all, of the daunorubicin glycosides accumulated in the experiments with production strain C5 derived from epsilon-rhodomycinone. A pathway network for the biosynthesis of daunorubicin glycosides is proposed that is in agreement with these data. In this proposed pathway network, epsilon-rhodomycinone is an intermediate in one of at least two pathways which yield daunorubicin glycosides. Images PMID:7425613

  19. 2006 Nature Publishing Group Structure of epsilon15 bacteriophage reveals

    E-print Network

    Jiang, Wen

    #12;© 2006 Nature Publishing Group Structure of epsilon15 bacteriophage reveals genome organization bacteriophage epsilon15 (ref. 1) determined from single-particle cryo-electron microscopy, without icosahedralDNA) bacteriophages are vectors for gene transfer among enteric bacteria, including important human pathogens7

  20. Acute hyperkalemia associated with intravenous epsilon-aminocaproic acid therapy

    Microsoft Academic Search

    Mark A. Perazella; Purba Biswas

    1999-01-01

    Epsilon-aminocaproic acid (Amicar) is used to treat severe hemorrhage refractory to usual medical management. This antifibrinolytic drug has been associated with a number of renal complications. However, there are no descriptions of this medication causing hyperkalemia. This report describes the development of hyperkalemia in a patient with underlying chronic renal insufficiency treated with intravenous epsilon-aminocaproic acid. The patient, who underwent

  1. Perturbative matching of the staggered four-fermion operators for {epsilon}'/{epsilon}

    SciTech Connect

    Lee, Weonjong

    2001-09-01

    Using staggered fermions, we calculate the perturbative corrections to the bilinear and four-fermion operators that are used in the numerical study of weak matrix elements for {epsilon}'/{epsilon}. We present results for one-loop matching coefficients between continuum operators, calculated in the naive dimensional regularization (NDR) scheme, and gauge invariant staggered fermion operators. In particular, we concentrate on Feynman diagrams of the current-current insertion type. We also present results for the tadpole improved operators. These results, combined with existing results for penguin diagrams, provide a complete one-loop renormalization of the staggered four-fermion operators. Therefore, using our results, it is possible to match a lattice calculation of K{sup 0}-{bar K}{sup 0} mixing and K{yields}{pi}{pi} decays to the continuum NDR results with all corrections of O(g{sup 2}) included.

  2. Comparative proteomic analysis of Clostridium difficile 

    E-print Network

    Chilton, Caroline Hazel

    2011-11-25

    The recent increase in availability of next generation sequencing methodologies has led to extensive analysis of the genome of Clostridium difficile. In contrast, protein expression analysis, crucial to the elucidation ...

  3. Recent advances in germination of Clostridium spores.

    PubMed

    Olguín-Araneda, Valeria; Banawas, Saeed; Sarker, Mahfuzur R; Paredes-Sabja, Daniel

    2014-08-15

    Members of Clostridium genus are a diverse group of anaerobic spore-formers that includes several pathogenic species. Their anaerobic requirement enhances the importance of the dormant spore morphotype during infection, persistence and transmission. Bacterial spores are metabolically inactive and may survive for long times in the environment and germinate in presence of nutrients termed germinants. Recent progress with spores of several Clostridium species has identified the germinant receptors (GRs) involved in nutrient germinant recognition and initiation of spore germination. Signal transduction from GRs to the downstream effectors remains poorly understood but involves the release of dipicolinic acid. Two mechanistically different cortex hydrolytic machineries are present in Clostridium spores. Recent studies have also shed light into novel biological events that occur during spore formation (accumulation of transcriptional units) and transcription during early spore outgrowth. In summary, this review will cover all of the recent advances in Clostridium spore germination. PMID:25132133

  4. Plasmids in Clostridium botulinum and related Clostridium species.

    PubMed Central

    Strom, M S; Eklund, M W; Poysky, F T

    1984-01-01

    Toxigenic Clostridium botulinum and nontoxigenic C. sporogenes, C. subterminale, and C. botulinum-like organisms from a variety of sources were screened for plasmids. Of the 68 toxigenic C. botulinum isolates, 56% carried one or more plasmids, ranging in mass from 2.1 to 81 megadaltons. Within individual groups (based on the type of neurotoxin produced), many strains showed identical plasmid banding patterns on agarose gels. Of the 15 nontoxigenic strains tested, 40% also carried one or more plasmids ranging from 1.7 to 25.0 megadaltons, with both unique and common banding patterns represented. A total of 67 plasmids from both toxigenic and nontoxigenic strains were detected. At this time, no phenotypic functions have been assessed for these plasmids, and they must therefore be considered cryptic. A variety of lysing and extraction techniques were necessary to detect plasmids in the different C. botulinum groups. Images PMID:6391384

  5. Vancomycin-resistant Clostridium innocuum bacteremia following oral vancomycin for Clostridium difficile infection.

    PubMed

    Hung, Yuan-Pin; Lin, Hsiao-Ju; Wu, Chi-Jung; Chen, Po-Lin; Lee, Jen-Chieh; Liu, Hsiao-Chieh; Wu, Yi-Hui; Yeh, Fang Hao; Tsai, Pei-Jane; Ko, Wen-Chien

    2014-12-01

    An 85 year-old male initially admitted for septic shock due to urinary tract infection experienced Clostridium difficile-associated diarrhea during hospitalization and was treated by oral vancomycin. His clinical course was complicated by cytomegalovirus colitis and then vancomycin-resistant Clostridium innocuum bacteremia, which was cured by uneventfully parenteral piperacillin-tazobactam therapy. PMID:25102472

  6. 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

  7. 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

  8. Clostridium difficile in paediatric populations.

    PubMed

    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

  9. Clostridium difficile the hospital plague.

    PubMed

    Czepiel, J; Kozicki, M; Panasiuk, P; Birczy?ska, M; Garlicki, A; Wese?ucha-Birczy?ska, A

    2015-03-16

    Clostridium difficile infection (CDI) has become one of the major public health threats in the last two decades. An increase has been observed not only in the rate of CDI, but also in its severity and mortality. Symptoms caused by this pathogen are accompanied by intense local and systemic inflammation. We confirmed that Raman microspectroscopy can help us in understanding CDI pathogenesis. A single erythrocyte of patients with CDI shows a difference, approximately 10 times, in the intensity of the Raman spectra at the beginning of hospitalization and after one week of treatment. The intensity level is an indicator of the spread of the inflammation within the cell, confirmed by standard laboratory tests. Many of the observed bands with enormously enhanced intensity, e.g. 1587, 1344, 1253, 1118 and 664 cm(-1), come from the symmetric vibration of the pyrrole ring. Heme variation of recovered cells in the acute CDI state between the first and the seventh day of treatment seems to show increased levels of oxygenated hemoglobin. Intense inflammation alters the conformation of the protein which is reflected in the significant changes in the amide I, II and III bands. There is an observed shift and a significant intensity increase of 1253 and 970 cm(-1) amide III and skeletal protein backbone CC stretching vibration bands, respectively. Principal Component Analysis (PCA) was used to find the variance in the data collected on the first and seventh day. PC2 loading in the 1645-1500 cm(-1) range shows an increase of heme, Tyr, Trp, or Phe vibrations because of changes in the protein microenvironment due to their exposure. Positive maxima at 1621, 1563 and 1550 in the PC2 loading originated from the ring vibrations. These observations indicate that Clostridium difficile toxins induce cytopathogenicity by altering cellular proteins. PMID:25627751

  10. Clostridium Difficile Infections - Multiple Languages: MedlinePlus

    MedlinePLUS

    ... sharing features on this page, please enable JavaScript. Clostridium Difficile Infections - Multiple Languages Arabic (???????) Spanish (español) Arabic ( ... Multimedia Patient Education Institute Spanish (español) Infecciones por Clostridium difficile Date last updated: 17 July 2012 Characters not ...

  11. The September epsilon Perseids in 2013

    NASA Astrophysics Data System (ADS)

    Gajdoš, Štefan; Tóth, Juraj; Kornoš, Leonard; Koukal, Jakub; Piffl, Roman

    2014-04-01

    An unexpected high activity (outburst) of the meteor shower September epsilon Perseids (SPE) was observed on 2013 September 9/10. The similar event occurred in 2008. We analysed SPE meteors observed in a frame of the European stations network (EDMONd) and collected in the video meteor orbits database EDMOND. Also, we compared two AMOS all-sky video observations of SPE meteors, performed at the Astronomical and Geophysical Observatory in Modra (AGO) and Arboretum in Tesarske Mlynany (ARBO) stations of the Slovak Video Meteor Network (SVMN). We obtained activity profiles of the 2013 SPE outburst during four hours around its maximum. Along with SPE activity profiles binned at 10 minutes for single-station meteors, we gained orbital characteristics of SPE meteors observed during the outburst, as well as a mean orbits of the SPE meteor stream in interval 2001-2012. The SPE outburst was confirmed by radio forward-scatter observations as well. The obtained observational results might be the starting point for modeling and explanation of SPE outbursts.

  12. 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.

  13. 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.

  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. Switchgrass (Panicum virgatum) fermentation by sequential culture of Clostridium thermocellum and Clostridium beijerinckii: effect of particle size on gas production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fuel alcohols can be produced by fermenting cellulosic biomass. Clostridium beijerinckii produces both ethanol and butanol, but it is non-cellulolytic. Cellulose requires saccharification prior to fermentation by C. beijerinckii. In contrast, the thermophile, Clostridium thermocellum, is highly ce...

  16. Clostridium hastiforme is a later synonym of Tissierella praeacuta

    E-print Network

    Bae, Jin-Woo

    priority. An emended description of the genus Tissierella is also given. Clostridium hastiforme MacClostridium hastiforme is a later synonym of Tissierella praeacuta Jin-Woo Bae,1 Ja Ryeong Park,1 species Clostridium hastiforme and Tissierella praeacuta appear to be similar from their published

  17. Cloning and characterization of the cytotoxin L-encoding gene of clostridium sordellii: homology with clostridium difficile cytotoxin B

    Microsoft Academic Search

    Gaynor A. Green; Véronique Schué; Henri Monteil

    1995-01-01

    Hybridization of an oligodeoxyribonucleotide (oligo) probe, designed from a repeated sequence ('oligo rep') at the C terminus of the Clostridium difficile (Cd) cytotoxin (Cyt), revealed that homologies exist between the Cd cyt gene and the genomes of several other clostridia, including Clostridium sordellii (Cs), suggesting a common ancestral cyt amongst the Clostridium genus. This Cd ‘oligo rep’ probe was used

  18. Quenched penguins, the Delta I=1/2 rule, and epsilon'/epsilon

    E-print Network

    Maarten Golterman; Elisabetta Pallante

    2006-09-30

    The transformation properties of strong penguin operators under the action of the flavor group change when they are considered as operators in (partially) quenched QCD instead of the unquenched theory. An ambiguity arises, which is parameterized by new low-energy constants in the effective theory describing non-leptonic kaon decays in the (partially) quenched setting. Here we summarize results of the analysis for the complete set of three-flavor strong penguin operators, consisting of products of two left-handed flavor currents, or a left- and a right-handed current. Our results imply that (partially) quenched lattice computations of the Delta I=1/2 rule and epsilon'/epsilon are both affected by ambiguities intrinsic to the use of the quenched approximation at leading order in the chiral expansion. The only exception is the partially quenched case with three light sea quarks, consistent with general expectations. We also address the issue of quenched ambiguities in the case of an active charm, correcting and extending that in Phys. Rev. D 74, 014509 (2006).

  19. 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

  20. Alzheimer pathology of patients carrying apolipoprotein E epsilon 4 allele.

    PubMed

    Heinonen, O; Lehtovirta, M; Soininen, H; Helisalmi, S; Mannermaa, A; Sorvari, H; Kosunen, O; Paljärvi, L; Ryynänen, M; Riekkinen, P J

    1995-01-01

    A recent report suggested that brains of Alzheimer patients homozygous for APOE epsilon 4 show increased amyloid pathology compared to APOE epsilon 3 homozygotes. We studied APOE allele frequencies in 73 AD patients and 38 controls. We also investigated relation of APOE genotypes to beta/A4 immunopositive plaques, cerebrovascular beta/A4 deposition, neurons expressing paired helical filaments (PHFs), and synaptophysin-like immunopositivity in 22 neuropathologically verified AD patients. We also correlated APOE genotypes of definite AD patients to beta/A4 immunoreactivity in dermal vessel walls detected in lifetime skin biopsy samples. APOE allele epsilon 4 frequency was increased in AD compared to nondemented controls (0.37 vs. 0.11; p = 0.006). The number of beta/A4 immunoreactive plaques, PHFs-containing neurons, the degree of cerebrovascular beta/A4 deposition or synaptophysin-like immunoreactivity did not differ significantly in AD patients with or without epsilon 4. beta/A4 deposition in dermal vessel walls was more frequent in definite AD patients with epsilon 4 (43%) than in patients without epsilon 4 (22%). However, the difference did not reach the statistical significance. PMID:8544899

  1. Thermostable chaperonin from Clostridium thermocellum.

    PubMed Central

    Cross, S J; Ciruela, A; Poomputsa, K; Romaniec, M P; Freedman, R B

    1996-01-01

    Homologues of the chaperonins Cpn60 and Cpn10 have been purified from the Gram-positive cellulolytic thermophile Clostridium thermocellum. The Cpn60 protein was purified by ATP-affinity chromatography and the Cpn10 protein was purified by gel-filtration, ion-exchange and hydrophobic interaction chromatographies. The identities of the proteins were confirmed by N-terminal sequence analysis and antigenic cross-reactivity. The Cpn60 homologue is a weak, thermostable ATPase (t1/2 at 70 decrees C more than 90 min) with optimum activity (Kcat 0.07 S-1) between 60 degrees C and 70 degrees C. The ATPase activity of the authentic Cpn60 was inhibited by Escherichia coli GroES. The catalytic properties of a recombinant C. thermocellum Cpn60 purified from a GST-Cpn60 fusion protein expressed in E. coli [Ciruela (1995) Ph.D. Thesis, University of Kent] were identical with those of the authentic C. thermocellum Cpn60. Gel-filtration studies show that at room temperature the Cpn60 migrates mainly as a heptamer. Electron microscopy confirms the presence of complexes showing 7-fold rotational symmetry and also reveals a small number of particles that seem to be tetradecamers with a similar structure to E. coli GroEL complexes. PMID:8687408

  2. Epidemiology of Clostridium difficile infection.

    PubMed

    Depestel, Daryl D; Aronoff, David M

    2013-10-01

    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 United States and Europe suggest that the incidence of CDI may have reached a crescendo in the recent years and is perhaps beginning to plateau. The acute care direct costs of CDI were estimated to be US$4.8 billion in 2008. However, nearly all the published studies have focused on CDI diagnosed and treated in the 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, to identify populations at risk, and to characterize the molecular epidemiology of strains causing CDI. PMID:24064435

  3. 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

  4. Hydrophobicity of Bacillus and Clostridium spores.

    PubMed Central

    Wiencek, K M; Klapes, N A; Foegeding, P M

    1990-01-01

    The hydrophobicities of spores and vegetative cells of several species of the genera Bacillus and Clostridium were measured by using the bacterial adherence to hexadecane assay and hydrophobic interaction chromatography. Although spore hydrophobicity varied among species and strains, the spores of each organism were more hydrophobic than the vegetative cells. The relative hydrophobicities determined by the two methods generally agreed. Sporulation media and conditions appeared to have little effect on spore hydrophobicity. However, exposure of spore suspensions to heat treatment caused a considerable increase in spore hydrophobicity. The hydrophobic nature of Bacillus and Clostridium spores suggests that hydrophobic interactions may play a role in the adhesion of these spores to surfaces. PMID:2275528

  5. Xylose fermentation with Clostridium thermohydrosulfuricum

    SciTech Connect

    Mancuso, A.; Wilke, C.R.; Blanch, H.W.

    1982-12-01

    In this study, the fermentation of xylose to ethanol with a thermophilic, strictly anaerobic bacterium, Clostridium thermohydrosulfuricum, was examined. The focus of this investigation was on the physiological parameters which most strongly affect the economic feasibility of using this bacterium for industrial ethanol production. In rich medium (containing economically impractical concentrations of yeast extract) yields as high as 0.43 gm ethanol/gm xylose and growth rates of 0.4 to 0.5 hr/sup -1/ were observed. The predominant by-products of the fermentation were acetate and lactate. Nutritional studies indicated that the cost of the growth medium could be dramatically reduced by replacing most of the yeast extract used with nicotinic acid and vitamin B/sup 12/. Ethanol was found to be very inhibitory to growth and ethanol formation. To overcome the problem of inhibition, cells were gradually adapted to high concentrations (up to 4.2%) of ethanol. However, the ethanol yield of adapted cells was typically 30 to 40% less than the yield of non-adapted cells. Environmental parameters such as pH and by-product concentrations had only a slight effect on the ethanol yield produced by tolerant cells. A mutant, selected from an adapted strain, was found to produce 60% less lactate than its parent. This low-lactate producing mutant had a slightly improved ethanol yield. The results obtained with the tolerant, low-lactate producing mutant were used in the design of an industrial-scale fermentation process. An economic evaluation of the process indicates that ethanol production with this bacterium is currently uneconomical.

  6. Atypical Glycolysis in Clostridium thermocellum

    PubMed Central

    Zhou, Jilai; Olson, Daniel G.; Argyros, D. Aaron; Deng, Yu; van Gulik, Walter M.; van Dijken, Johannes P.

    2013-01-01

    Cofactor specificities of glycolytic enzymes in Clostridium thermocellum were studied with cellobiose-grown cells from batch cultures. Intracellular glucose was phosphorylated by glucokinase using GTP rather than ATP. Although phosphofructokinase typically uses ATP as a phosphoryl donor, we found only pyrophosphate (PPi)-linked activity. Phosphoglycerate kinase used both GDP and ADP as phosphoryl acceptors. In agreement with the absence of a pyruvate kinase sequence in the C. thermocellum genome, no activity of this enzyme could be detected. Also, the annotated pyruvate phosphate dikinase (ppdk) is not crucial for the generation of pyruvate from phosphoenolpyruvate (PEP), as deletion of the ppdk gene did not substantially change cellobiose fermentation. Instead pyruvate formation is likely to proceed via a malate shunt with GDP-linked PEP carboxykinase, NADH-linked malate dehydrogenase, and NADP-linked malic enzyme. High activities of these enzymes were detected in extracts of cellobiose-grown cells. Our results thus show that GTP is consumed while both GTP and ATP are produced in glycolysis of C. thermocellum. The requirement for PPi in this pathway can be satisfied only to a small extent by biosynthetic reactions, in contrast to what is generally assumed for a PPi-dependent glycolysis in anaerobic heterotrophs. Metabolic network analysis showed that most of the required PPi must be generated via ATP or GTP hydrolysis exclusive of that which happens during biosynthesis. Experimental proof for the necessity of an alternative mechanism of PPi generation was obtained by studying the glycolysis in washed-cell suspensions in which biosynthesis was absent. Under these conditions, cells still fermented cellobiose to ethanol. PMID:23435896

  7. Clostridium difficile phages: still difficult?

    PubMed Central

    Hargreaves, Katherine R.; Clokie, Martha R. J.

    2014-01-01

    Phages that infect Clostridium difficile were first isolated for typing purposes in the 1980s, but their use was short lived. However, the rise of C. difficile epidemics over the last decade has triggered a resurgence of interest in using phages to combat this pathogen. Phage therapy is an attractive treatment option for C. difficile infection, however, developing suitable phages is challenging. In this review we summarize the difficulties faced by researchers in this field, and we discuss the solutions and strategies used for the development of C. difficile phages for use as novel therapeutics. Epidemiological data has highlighted the diversity and distribution of C. difficile, and shown that novel strains continue to emerge in clinical settings. In parallel with epidemiological studies, advances in molecular biology have bolstered our understanding of C. difficile biology, and our knowledge of phage–host interactions in other bacterial species. These three fields of biology have therefore paved the way for future work on C. difficile phages to progress and develop. Benefits of using C. difficile phages as therapeutic agents include the fact that they have highly specific interactions with their bacterial hosts. Studies also show that they can reduce bacterial numbers in both in vitro and in vivo systems. Genetic analysis has revealed the genomic diversity among these phages and provided an insight into their taxonomy and evolution. No strictly virulent C. difficile phages have been reported and this contributes to the difficulties with their therapeutic exploitation. Although treatment approaches using the phage-encoded endolysin protein have been explored, the benefits of using “whole-phages” are such that they remain a major research focus. Whilst we don’t envisage working with C. difficile phages will be problem-free, sufficient study should inform future strategies to facilitate their development to combat this problematic pathogen. PMID:24808893

  8. Atypical glycolysis in Clostridium thermocellum.

    PubMed

    Zhou, Jilai; Olson, Daniel G; Argyros, D Aaron; Deng, Yu; van Gulik, Walter M; van Dijken, Johannes P; Lynd, Lee R

    2013-05-01

    Cofactor specificities of glycolytic enzymes in Clostridium thermocellum were studied with cellobiose-grown cells from batch cultures. Intracellular glucose was phosphorylated by glucokinase using GTP rather than ATP. Although phosphofructokinase typically uses ATP as a phosphoryl donor, we found only pyrophosphate (PPi)-linked activity. Phosphoglycerate kinase used both GDP and ADP as phosphoryl acceptors. In agreement with the absence of a pyruvate kinase sequence in the C. thermocellum genome, no activity of this enzyme could be detected. Also, the annotated pyruvate phosphate dikinase (ppdk) is not crucial for the generation of pyruvate from phosphoenolpyruvate (PEP), as deletion of the ppdk gene did not substantially change cellobiose fermentation. Instead pyruvate formation is likely to proceed via a malate shunt with GDP-linked PEP carboxykinase, NADH-linked malate dehydrogenase, and NADP-linked malic enzyme. High activities of these enzymes were detected in extracts of cellobiose-grown cells. Our results thus show that GTP is consumed while both GTP and ATP are produced in glycolysis of C. thermocellum. The requirement for PPi in this pathway can be satisfied only to a small extent by biosynthetic reactions, in contrast to what is generally assumed for a PPi-dependent glycolysis in anaerobic heterotrophs. Metabolic network analysis showed that most of the required PPi must be generated via ATP or GTP hydrolysis exclusive of that which happens during biosynthesis. Experimental proof for the necessity of an alternative mechanism of PPi generation was obtained by studying the glycolysis in washed-cell suspensions in which biosynthesis was absent. Under these conditions, cells still fermented cellobiose to ethanol. PMID:23435896

  9. 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.

  10. Clostridium difficile-Associated Diarrhea and Colitis

    Microsoft Academic Search

    Dale N. Gerding; Stuart Johnson; Lance R. Peterson; Maury E. Mulligan; Joseph Silva Jr.

    1995-01-01

    Objectives: To review and summarize the status of diagnosis, epidemiology, infection control, and treatment of Clostridium difficile-associated disease (CDAD). Diagnosis: A case definition of CDAD should include the presence of symptoms (usually diarrhea) and at least one of the following positive tests: endoscopy revealing pseudomembranes, stool cytotoxicity test for toxin B, stool enzyme immunoassay for toxin A or B, or

  11. 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

  12. Clostridium difficile in poultry and poultry meat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The incidence and severity of disease associated with toxigenic Clostridium difficile have increased in hospitals in North America from the emergence of newer, more virulent strains. Toxigenic C. difficile has been isolated from food animals and retail meat with potential implications of transfer t...

  13. Clostridium difficile in seafood and fish.

    PubMed

    Metcalf, Devon; Avery, Brent P; Janecko, Nicol; Matic, Nancy; Reid-Smith, Richard; Weese, J Scott

    2011-04-01

    The objective of this study was to determine the prevalence of Clostridium difficile contamination in retail seafood and fish from Canadian grocery stores. C. difficile was found in 4.8% (5/119) of the samples. This study, combined with studies of other food sources, suggests that widespread contamination of food is common. PMID:21376822

  14. Clostridium septicum Empyema in an Immunocompetent Woman.

    PubMed

    Granok, Alexander B; Mahon, Patrick A; Biesek, Genesio W

    2010-01-01

    We report a case of a Clostridium septicum empyema in an immunocompetent woman following operation for an incarcerated internal hernia. The patient was successfully treated with pleural decortication and an extended course of postoperative antibiotics. This is the first report of such an infection in the medical literature. PMID:20490275

  15. Clostridium botulinum in Cattle and Dairy Products

    Microsoft Academic Search

    Miia Lindström; Jan Myllykoski; Seppo Sivelä; Hannu Korkeala

    2010-01-01

    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

  16. A modular system for Clostridium shuttle plasmids

    Microsoft Academic Search

    John T. Heap; Oliver J. Pennington; Stephen T. Cartman; Nigel P. Minton

    2009-01-01

    Despite their medical and industrial importance, our basic understanding of the biology of the genus Clostridium is rudimentary in comparison to their aerobic counterparts in the genus Bacillus. A major contributing factor has been the comparative lack of sophistication in the gene tools available to the clostridial molecular biologist, which are immature, and in clear need of development. The transfer

  17. Sigma factor and sporulation genes in Clostridium

    Microsoft Academic Search

    Uwe Sauer; Joseph D. Santangelo; Anke Treuner; Malte Buchholz; Peter Dürre

    1995-01-01

    The genus Clostridium, represented by Gram-positive, anaerobic, spore-forming bacteria, is well known for its clinical importance and considerable biotechnological potential. Recently, evidence for a functional role of the transcription factors ?A, ?E, ?G, and ?K in this genus was provided by cloning and sequencing these genes from C. acetobutylicum. In C. kluyveri, a partially sequenced open reading frame was found

  18. Cellulolytic and physiological properties of Clostridium thermocellum

    Microsoft Academic Search

    T. K. Ng; P. J. Weimer; J. G. Zeikus

    1977-01-01

    Three strains of Clostridium thermocellum obtained from various sources were found to have nearly identical deoxyribonucleic acid guanosine plus cytosine contents that ranged from 38.1–39.5 mole-%. All strain examined fermented only cellulose and cellulose derivatives, but not glucose, or xylose or other sugars. The principal cellulose fermentation products were ethanol, lactate, acetate, hydrogen and carbon dioxide. Growth of C. thermocellum

  19. Effects of probiotic, Clostridium butyricum, on growth performance, immune function, and cecal microflora in broiler chickens.

    PubMed

    Yang, C M; Cao, G T; Ferket, P R; Liu, T T; Zhou, L; Zhang, L; Xiao, Y P; Chen, A G

    2012-09-01

    Four hundred and fifty 1-d-old male Lingnan Yellow broiler chickens were used to investigate the effects of Clostridium butyricum on growth performance, immune function, and cecal microflora. The birds were randomly assigned to 5 treatments and offered the same antibiotic-free basal diets for 42 d. The treatments were as follows: no addition (control), 1 × 10(7) cfu C. butyricum/kg of diet (CB1), 2 × 10(7) cfu C. butyricum/kg of diet (CB2), 3 × 10(7) cfu C. butyricum/kg of diet (CB3), and 10 mg of colistine sulfate/kg of diet (antibiotic). Birds fed either CB2 or antibiotic had greater overall BW than those in the control group. During d 1 to 7, d 21 to 42, and d 1 to 42, birds fed either CB2 or CB3 or the antibiotic diet had greater ADG compared with those in the control group. No significant differences were observed in BW or ADG among the CB2, CB3, and antibiotic groups. Birds fed the CB2 or CB3 diet had greater concentrations of IgA and IgG in the serum from d 14 to 42 and greater IgM in the serum from d 21 to 42 than those in the control group. Birds fed the CB3 diet had a greater concentration of complement component 3 in the serum than those in the control group from d 7 to 42. Dietary C. butyricum decreased (P < 0.05) Escherichia coli in cecal contents on d 14 and 42, and both CB2 and CB3 decreased (P < 0.05) cecal Salmonella and Clostridium perfringen from d 14 to 42 compared with the control. Broilers fed either CB2 or CB3 had greater cecal Lactobacillus and Bifidobacterium counts from d 21 to 42, and birds fed C. butyricum had greater cecal C. butyricum counts during the whole period compared with those in the control group. The results indicate that C. butyricum promotes growth performance and immune function and benefits the balance of the intestinal microflora in broiler chickens. PMID:22912445

  20. Bacteriostatic effect of orally administered bovine lactoferrin on proliferation of Clostridium species in the gut of mice fed bovine milk.

    PubMed Central

    Teraguchi, S; Shin, K; Ozawa, K; Nakamura, S; Fukuwatari, Y; Tsuyuki, S; Namihira, H; Shimamura, S

    1995-01-01

    When milk-fed mice were orally inoculated with Clostridium ramosum C1, this strain proliferated in the gut and became the dominant component of the fecal microflora. In this experimental model, bovine lactoferrin (bLF) administered with milk suppressed the proliferation of this strain in vivo and decreased the numbers of C. ramosum and other bacteria in the feces. This bacteriostatic effect of bLF was dependent on the concentration of bLF, the duration of feeding, and the administered dose of C. ramosum C1. Compared with bovine serum albumin, ovalbumin, bovine whey protein isolate, or bovine casein, only bLF showed this specific activity. A similar effect of bLF was observed after oral inoculation with C. ramosum JCM 1298, C. paraputrificum VPI 6372, or C. perfringens ATCC 13124. A hydrolysate prepared by digestion of bLF with porcine pepsin showed the same inhibitory effect on proliferation of C. ramosum in vivo as occurred with undigested bLF. These results indicate that ingested bLF can exert a bacteriostatic effect against clostridia in the gut even after it has been digested to some extent. PMID:7574587

  1. Epsilon-Unfolding Orthogonal Polyhedra Mirela Damian1

    E-print Network

    O'Rourke, Joseph

    Epsilon-Unfolding Orthogonal Polyhedra Mirela Damian1 , Robin Flatland2 , Joseph O'Rourke3 1 Dept to be as thin as = 1/2(n) . Key words. general unfolding, grid unfolding, orthogonal polyhedra, genus- zero 1 places no restriction on the cuts. General unfoldings are known for convex polyhedra

  2. Eutectic epsilon-near-zero metamaterial terahertz waveguides

    E-print Network

    Eutectic epsilon-near-zero metamaterial terahertz waveguides M. Massaouti,1 A. A. Basharin,1,2 M phenomena of enhanced THz transmission through a subwavelength LiF dielectric rod lattice embedded that subwavelength waveguiding of terahertz radiation is achieved within an alkali­halide eutectic metamaterial

  3. 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.

  4. 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.

  5. Providing a Virtual Initiation for Epsilon Pi Tau

    ERIC Educational Resources Information Center

    Sanders, Craig S.; Griffin, Kathryn M.

    2005-01-01

    One of the requirements of the Epsilon Pi Tau (EPT) initiation is the apprentice has to physically be at the initiation (EPT, 2004). Since the majority of nontraditional students and working professionals are physically removed from an initiation site, they have missed the opportunity to join EPT. On 8 April, 2005, the Beta Mu Chapter of The…

  6. Infinite Bottles of Beer Mathematical Concepts with Epsilon Pain

    E-print Network

    Indiana University

    1 Infinite Bottles of Beer Mathematical Concepts with Epsilon Pain Or: A cantorial approach this: A hundred bottles of beer on the wall, a hundred bottles of beer; If one of those bottles should happen to fall, 99 bottles of beer on the wall. 99 bottles of beer on the wall, 99 bottles of beer

  7. 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

  8. Validated near-atomic resolution structure of bacteriophage epsilon15 derived from cryo-EM

    E-print Network

    Jiang, Wen

    Validated near-atomic resolution structure of bacteriophage epsilon15 derived from cryo important contribu- tions to modern structural biology. Bacteriophages, the most diverse and abundant previous electron cryomicroscopy structure of Salmonella bacteriophage epsilon15, achieving a resolution

  9. Complementarity between epsilon and phi sequences in pregenomic RNA influences hepatitis B virus replication efficiency.

    PubMed

    Oropeza, Claudia E; McLachlan, Alan

    2007-03-15

    Hepatitis B virus (HBV) replication requires the viral polymerase to reverse transcribe the 3.5-kb pregenomic viral RNA within the nucleocapsid. It has been proposed that a sequence element designated phi (phi), which is located 32 nucleotides upstream of the 3' DR1 pregenomic RNA sequence and is complementary to epsilon, is required for efficient minus-strand synthesis because it may mediate the translocation of the viral polymerase plus the three nucleotide primer from epsilon to DR1. A mutation in phi has been identified which can be compensated for with a complementary mutation in epsilon. This observation supports the suggestion that epsilon and phi base pair during the process of polymerase translocation from epsilon to DR1. However, additional mutations in phi were not complemented by the corresponding mutations in epsilon indicating that the functional recognition of epsilon and epsilon/phi stem-loop structures by polymerase probably requires both sequence- and structure-specific information. PMID:17056086

  10. Composition and primary structure of the F1F0 ATP synthase from the obligately anaerobic bacterium Clostridium thermoaceticum.

    PubMed Central

    Das, A; Ljungdahl, L G

    1997-01-01

    The subunit composition and primary structure of the proton-translocating F1F0 ATP synthase have been determined in Clostridium thermoaceticum. The isolated enzyme has a subunit composition identical to that of the F1F0 ATP synthase purified from Clostridium thermoautotrophicum (A. Das, D. M. Ivey, and L. G. Ljungdahl, J. Bacteriol. 179:1714-1720, 1997), both having six different polypeptides. The molecular masses of the six subunits were 60, 50, 32, 17, 19, and 8 kDa, and they were identified as alpha, beta, gamma, delta, epsilon, and c, respectively, based on their reactivity with antibodies against the F1 ATPase purified from C. thermoautotrophicum and by comparing their N-terminal amino acid sequences with that deduced from the cloned genes of the C. thermoaceticum atp operon. The subunits a and b found in many bacterial ATP synthases could not be detected either in the purified ATP synthase or crude membranes of C. thermoaceticum. The C. thermoaceticum atp operon contained nine genes arranged in the order atpI (i), atpB (a), atpE (c), atpF (b), atpH (delta), atpA (alpha), atpG (gamma), atpD (beta), and atpC (epsilon). The deduced protein sequences of the C. thermoaceticum ATP synthase subunits were comparable with those of the corresponding subunits from Escherichia coli, thermophilic Bacillus strain PS3, Rhodospirillum rubrum, spinach chloroplasts, and the cyanobacterium Synechococcus strain PCC 6716. The analysis of total RNA by Northern hybridization experiments reveals the presence of transcripts (mRNA) of the genes i, a, and b subunits not found in the isolated enzyme. Analysis of the nucleotide sequence of the atp genes reveals overlap of the structural genes for the i and a subunits and the presence of secondary structures (in the b gene) which could influence the posttranscriptional regulation of the corresponding genes. PMID:9171425

  11. 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

  12. Screening for plasmids in the genus Clostridium

    Microsoft Academic Search

    C.-K. Lee; P. Dürre; H. Hippe; G. Gottschalk

    1987-01-01

    A plasmid screening was performed on 150 strains out of 75 clostridial species using a modification of the alkaline-lysis procedure. In 26 strains representing 21 species one or more plasmid bands were detected ranging in size from 3 to more than 100 kilobase pairs. Clostridium aceticum proved to contain a single small plasmid (pCA1) of 5.4 kbp as revealed by

  13. Association of apolipoprotein E allele {epsilon}4 with late-onset sporadic Alzheimer`s disease

    SciTech Connect

    Lucotte, G.; David, F.; Berriche, S. [Regional Center of Neurogenetics, Reims (France)] [and others

    1994-09-15

    Apolipoprotein E, type {epsilon}4 allele (ApoE {epsilon}4), is associated with late-onset sporadic Alzheimer`s disease (AD) in French patients. The association is highly significant (0.45 AD versus 0.12 controls for {epsilon}4 allele frequencies). These data support the involvement of ApoE {epsilon}4 allele as a very important risk factor for the clinical expression of AD. 22 refs., 1 fig., 3 tabs.

  14. Draft Genome Sequence of Clostridium butyricum Strain NOR 33234, Isolated from an Elderly Patient with Diarrhea

    PubMed Central

    Kwok, Jamie S. L.; Ip, Margaret; Chan, Ting-Fung; Lam, Wai-Yip

    2014-01-01

    Clostridium butyricum is one of the species frequently present in patients’ stool samples. However, the identification of this species is sometimes difficult. Here, we present the draft genome of Clostridium butyricum NOR 33234, which was isolated from a patient with suspected Clostridium difficile infection-associated diarrhea and resembles Clostridium clostridioforme in biochemical tests. PMID:25540356

  15. The effect of probiotics on clostridium difficile diarrhea

    Microsoft Academic Search

    Mark Pochapin

    2000-01-01

    Clostridium difficile is the leading cause of nosocomially acquired intestinal infection in the United States, affecting virtually all cases of pseudomembranous colitis and up to 20% of cases of antibiotic-associated diarrhea. Even after receiving antibiotic treatment with either metronidazole or vancomycin, 20% of patients will have recurrent Clostridium difficile diarrhea. An innovative approach to the problem involves the introduction of

  16. 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

  17. BOWEL PREPARATION FOR COLECTOMY AND RISK OF Clostridium difficile INFECTION

    PubMed Central

    Krapohl, Greta L.; Phillips, Laurel; Campbell, Darrell A.; Hendren, Samantha; Banerjee, Mousumi; Metzger, Bonnie; Morris, Arden M.

    2011-01-01

    Background Mechanical bowel preparation prior to colectomy is controversial for several reasons, including a theoretically increased risk of Clostridium difficile infection. Objective To compare the incidence of Clostridium difficile infection among patients who underwent mechanical bowel preparation and those who did not. A secondary objective was to assess the association between Clostridium difficile infection and the use of oral antibiotics. Design Observational cohort study. Setting The Michigan Surgical Quality Collaborative Colectomy Project (n=24 hospitals) participates in the American College of Surgeons- National Surgical Quality Improvement Program with additional targeted data specific to colectomy patients. Patients Adult patients (21 years and older) admitted to participating hospitals for elective colectomy between August 2007 and June 2009. Main Outcome Measure Laboratory detection of a positive Clostridium difficile toxin assay or stool culture. Results 2263 patients underwent colectomy and fulfilled inclusion criteria. 54 developed a Clostridium difficile infection, for a hospital median rate of 2.8% (range 0 to 14.7%). Use of mechanical bowel preparation was not associated with increased incidence of Clostridium difficile infection (p=0.95). Among 1685 patients that received mechanical bowel preparation, 684 (41%) received oral antibiotics. The proportion of patients who were diagnosed with Clostridium difficile infection after using preoperative oral antibiotics was smaller than the proportion of patients with Clostridium difficile infection who did not receive oral antibiotics (1.6% vs. 2.9%, p=0.09). Limitations Potential underestimation of Clostridium difficile infection due to the study's strict data collection criteria and risk of undetected infection after postoperative day 30. Conclusions In contrast to previous single-center data, this multi-center study showed preoperative use of mechanical bowel preparation was not associated with increased risk of Clostridium difficile infection after colectomy. Moreover, the addition of oral antibiotics with mechanical bowel preparation did not confer any additional risk of infection. PMID:21654247

  18. Variation of the radial velocity of Epsilon Cygni A

    NASA Technical Reports Server (NTRS)

    Mcmillan, R. S.; Smith, P. H.; Moore, T. L.; Perry, M. L.

    1992-01-01

    The paper reports a series of 217 measurements of the radial velocity of Epsilon Cygni A made between May 16, 1987 and May 17, 1991 with an uncertainty per observation of +/- 12 m/s. The results indicate a sustained drift of -60 m/s yr. Lower limits on the companion's mass are presented as functions of period and primary mass. The companion is probably more massive than a planet.

  19. Theory of epsilon-near-zero modes in ultrathin films

    NASA Astrophysics Data System (ADS)

    Campione, Salvatore; Brener, Igal; Marquier, Francois

    2015-03-01

    The physics of the epsilon-near-zero (ENZ) mode, which is supported by a nanolayer at the frequency where the dielectric permittivity vanishes, has recently been a subject of debate. In this Rapid Communication, we thoroughly investigate and clarify the physics of this mode, providing its main characteristics and its domain of existence. This understanding will benefit all the applications that rely on ENZ modes in semiconductor nanolayers, including directional perfect absorption, voltage-tunable devices, and ultrafast thermal emission.

  20. Phenotypic Differentiation between Clostridium hast for me and Clostridium subterminale by Headspace Gas Chromatography

    Microsoft Academic Search

    PHILIPPE NIEL; ALAIN RIMBAULT; GENEVIEVE CAMPION; GEORGES LELUAN

    Bacteria of the genus Clostridium form a very heteroge- neous group (3), and their identification requires both con- ventional phenotypic tests and analysis of volatile fatty acids by gas chromatography (4). As in some instances these procedures are not sufficient to assign a strain to a no- menspecies and to differentiate some taxa, additional criteria are needed (2). The simultaneous

  1. Clostridium aldenense sp. nov. and Clostridium citroniae sp. nov. Isolated from Human Clinical Infections

    Microsoft Academic Search

    Yumi A. Warren; Kerin L. Tyrrell; Diane M. Citron; Ellie J. C. Goldstein

    2006-01-01

    One hundred eight isolates were previously identified in our laboratory as Clostridium clostridioforme by colonial and cellular morphology, as well as biochemical tests. Recent studies have indicated that there are actually three different species in this C. clostridioforme group: C. hathewayi, C. bolteae, and C. clostridioforme. Our isolates were reexamined using biochemical and enzymatic tests and molecular methods. Forty-six isolates

  2. MAGNETIC ACTIVITY CYCLES IN THE EXOPLANET HOST STAR {epsilon} ERIDANI

    SciTech Connect

    Metcalfe, T. S.; Mathur, S. [Space Science Institute, 4750 Walnut Street, Suite 205, Boulder, CO 80301 (United States); Buccino, A. P.; Mauas, P. J. D.; Petrucci, R. [Instituto de Astronomia y Fisica del Espacio (CONICET), C.C. 67 Sucursal 28, C1428EHA-Buenos Aires (Argentina); Brown, B. P. [Department of Astronomy and Center for Magnetic Self-Organization, University of Wisconsin, Madison, WI 53706-1582 (United States); Soderblom, D. R. [Space Telescope Science Institute, 3700 San Martin Dr., Baltimore, MD 21218 (United States); Henry, T. J. [Department of Physics and Astronomy, Georgia State University, Atlanta, GA 30302 (United States); Hall, J. C. [Lowell Observatory, 1400 West Mars Hill Road, Flagstaff, AZ 86001 (United States); Basu, S. [Department of Astronomy, Yale University, P.O. Box 208101, New Haven, CT 06520 (United States)

    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 Boehm-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.

  3. Plasmidome Interchange between Clostridium botulinum, Clostridium novyi and Clostridium haemolyticum Converts Strains of Independent Lineages into Distinctly Different Pathogens

    PubMed Central

    Skarin, Hanna; Segerman, Bo

    2014-01-01

    Clostridium botulinum (group III), Clostridium novyi and Clostridium haemolyticum are well-known pathogens causing animal botulism, gas gangrene/black disease, and bacillary hemoglobinuria, respectively. A close genetic relationship exists between the species, which has resulted in the collective term C. novyi sensu lato. The pathogenic traits in these species, e.g., the botulinum neurotoxin and the novyi alpha toxin, are mainly linked to a large plasmidome consisting of plasmids and circular prophages. The plasmidome of C. novyi sensu lato has so far been poorly characterized. In this study we explored the genomic relationship of a wide range of strains of C. novyi sensu lato with a special focus on the dynamics of the plasmidome. Twenty-four genomes were sequenced from strains selected to represent as much as possible the genetic diversity in C. novyi sensu lato. Sixty-one plasmids were identified in these genomes and 28 of them were completed. The genomic comparisons revealed four separate lineages, which did not strictly correlate with the species designations. The plasmids were categorized into 13 different plasmid groups on the basis of their similarity and conservation of plasmid replication or partitioning genes. The plasmid groups, lineages and species were to a large extent entwined because plasmids and toxin genes had moved across the lineage boundaries. This dynamic process appears to be primarily driven by phages. We here present a comprehensive characterization of the complex species group C. novyi sensu lato, explaining the intermixed genetic properties. This study also provides examples how the reorganization of the botulinum toxin and the novyi alpha toxin genes within the plasmidome has affected the pathogenesis of the strains. PMID:25254374

  4. Annotation of the Clostridium Acetobutylicum Genome

    SciTech Connect

    Daly, M. J.

    2004-06-09

    The genome sequence of the solvent producing bacterium Clostridium acetobutylicum ATCC824, has been determined by the shotgun approach. The genome consists of a 3.94 Mb chromosome and a 192 kb megaplasmid that contains the majority of genes responsible for solvent production. Comparison of C. acetobutylicum to Bacillus subtilis reveals significant local conservation of gene order, which has not been seen in comparisons of other genomes with similar, or, in some cases, closer, phylogenetic proximity. This conservation allows the prediction of many previously undetected operons in both bacteria.

  5. 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

  6. An Attempt to Derive the epsilon Equation from a Two-Point Closure

    NASA Technical Reports Server (NTRS)

    Canuto, V. M.; Cheng, Y.; Howard, A. M.

    2010-01-01

    The goal of this paper is to derive the equation for the turbulence dissipation rate epsilon for a shear-driven flow. In 1961, Davydov used a one-point closure model to derive the epsilon equation from first principles but the final result contained undetermined terms and thus lacked predictive power. Both in 1987 and in 2001, attempts were made to derive the epsilon equation from first principles using a two-point closure, but their methods relied on a phenomenological assumption. The standard practice has thus been to employ a heuristic form of the equation that contains three empirical ingredients: two constants, c(sub 1 epsilon), and c(sub 2 epsilon), and a diffusion term D(sub epsilon) In this work, a two-point closure is employed, yielding the following results: 1) the empirical constants get replaced by c(sub 1), c(sub 2), which are now functions of Kappa and epsilon; 2) c(sub 1) and c(sub 2) are not independent because a general relation between the two that are valid for any Kappa and epsilon are derived; 3) c(sub 1), c(sub 2) become constant with values close to the empirical values c(sub 1 epsilon), c(sub epsilon 2), (i.e., homogenous flows); and 4) the empirical form of the diffusion term D(sub epsilon) is no longer needed because it gets substituted by the Kappa-epsilon dependence of c(sub 1), c(sub 2), which plays the role of the diffusion, together with the diffusion of the turbulent kinetic energy D(sub Kappa), which now enters the new equation (i.e., inhomogeneous flows). Thus, the three empirical ingredients c(sub 1 epsilon), c(sub epsilon 2), D (sub epsilon)are replaced by a single function c(sub 1)(Kappa, epsilon ) or c(sub 2)(Kappa, epsilon ), plus a D(sub Kappa)term. Three tests of the new equation for epsilon are presented: one concerning channel flow and two concerning the shear-driven planetary boundary layer (PBL).

  7. Clostridium septicum alpha-toxin forms pores and induces rapid cell necrosis.

    PubMed

    Knapp, Oliver; Maier, Elke; Mkaddem, Sanae Ben; Benz, Roland; Bens, Marcelle; Chenal, Alexandre; Geny, Blandine; Vandewalle, Alain; Popoff, Michel R

    2010-01-01

    Alpha-toxin is the unique lethal virulent factor produced by Clostridium septicum, which causes traumatic or non-traumatic gas gangrene and necrotizing enterocolitis in humans. Here, we analyzed channel formation of the recombinant septicum alpha-toxin and characterized its activity on living cells. Recombinant septicum alpha-toxin induces the formation of ion-permeable channels with a single-channel conductance of about 175pS in 0.1M KCl in lipid bilayer membranes, which is typical for a large diffusion pore. Septicum alpha-toxin channels remained mostly in the open configuration, displayed no lipid specificity, and exhibited slight anion selectivity. Septicum alpha-toxin caused a rapid decrease in the transepithelial electrical resistance of MDCK cell monolayers grown on filters, and induced a rapid cell necrosis in a variety of cell lines, characterized by cell permeabilization to propidium iodide without DNA fragmentation and activation of caspase-3. Septicum alpha-toxin also induced a rapid K(+) efflux and ATP depletion. Incubation of the cells in K(+)-enriched medium delayed cell death caused by septicum alpha-toxin or epsilon-toxin, another potent pore-forming toxin, suggesting that the rapid loss of intracellular K(+) represents an early signal of pore-forming toxins-mediated cell necrosis. PMID:19632260

  8. Key research issues in Clostridium difficile

    PubMed Central

    Zhanel, George; Hammond, Greg

    2005-01-01

    Clostridium difficile is an emerging pathogen that causes C difficile-associated diarrhea, an important nosocomial infection. Control of this infection remains a challenge, and much needs to be determined about the antimicrobial resistance of the organism, antibiotic stewardship, contamination of the patient environment, and various host factors that determine susceptibility or resistance to infection. A national symposium focusing on C difficile infections, the Clostridium difficile Symposium on Emerging Issues and Research, was hosted on November 23, 2004, by the Department of Medical Microbiology and Infectious Diseases at the University of Manitoba, Winnipeg, Manitoba, in partnership with the Canadian Institutes of Health Research. This symposium, which aimed to summarize key research issues regarding C difficile infections in Canada, had the following objectives: to provide a forum for learning and discussion about C difficile and its impact on the health of Canadians; to identify the key research issues that should be addressed; and to explore potential research funding opportunities and collaboration. The present report summarizes key research issues identified for C difficile infections in Canada by addressing four major themes: diagnosis and surveillance, infection prevention and control, antibiotic stewardship, and clinical management. PMID:18159559

  9. 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.

  10. Detecting a rotation in the epsilon Eridani debris disc

    E-print Network

    C. J. Poulton; J. S. Greaves; A. C. Cameron

    2006-06-23

    The evidence for a rotation of the epsilon Eridani debris disc is examined. Data at 850 micron wavelength were previously obtained using the Submillimetre Common User Bolometer Array (SCUBA) over periods in 1997-1998 and 2000-2002. By chi-square fitting after shift and rotation operations, images from these two epochs were compared to recover proper motion and orbital motion of the disc. The same procedures were then performed on simulated images to estimate the accuracy of the results. Minima in the chi-square plots indicate a motion of the disc of approximately 0.6'' per year in the direction of the star's proper motion. This underestimates the true value of 1'' per year, implying that some of the structure in the disc region is not associated with epsilon Eridani, originating instead from background galaxies. From the chi-square fitting for orbital motion, a counterclockwise rotation rate of ~2.75 degrees per year is deduced. Comparisons with simulated data in which the disc is not rotating show that noise and background galaxies result in approximately Gaussian fluctuations with a standard deviation +/-1.5 degrees per year. Thus counterclockwise rotation of disc features is supported at approximately a 2-sigma level, after a 4-year time difference. This rate is faster than the Keplerian rate of 0.65 degrees per year for features at ~65 AU from the star, suggesting their motion is tracking a planet inside the dust ring. Future observations with SCUBA-2 can rule out no rotation of the epsilon Eridani dust clumps with ~4-sigma confidence. Assuming a rate of about 2.75 degrees per year, the rotation of the features after a 10-year period could be shown to be >1 degree per year at the 3-sigma level.

  11. Gearing up for Epsilon Aurigae's First Eclipse of the Millennium

    NASA Astrophysics Data System (ADS)

    Hopkins, Jeffery L.; Schanne, Lothar; Stencel, Robert E.

    2008-05-01

    The mysterious 3rd magnitude long period eclipsing binary star system, epsilon Aurigae, is predicted to start its two-year eclipse in the July 2009. This may be when the real excitement starts but much is to be learned before first contact. This paper will discuss current observational results that have accumulated data using photometry, spectroscopy and other data sources. While the system is ideal for single channel photometry, due to the system brightness and distant comparison star, CCD photometry presents some interesting challenges. A fairly simple way for amateur astronomers to do BVRI CCD photometry of the system is using a 50 mm camera lens and DSI Pro camera is discussed.

  12. Epsilon-near-zero metamaterials for tailoring ultrashort pulses

    NASA Astrophysics Data System (ADS)

    Zhai, Tianrui; Zhang, Xinping

    2013-11-01

    Interaction between epsilon-near-zero (ENZ) metamaterials and optical pulses is investigated theoretically. A transform between total transmission and total reflection is revealed for the ultrashort pulses incident onto an ENZ slab, which can be controlled by changing the thickness of the slab. Furthermore, the resonance of the ENZ materials leads to a cutoff frequency in spectroscopic response, where longer wavelengths have no chance to propagate through the ENZ materials and are totally reflected. These characteristics enable the ENZ materials to be applied in ultrashort pulse shapers, cutoff filters, and Fabry-Pérot resonators.

  13. 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.

  14. 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.

  15. 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.

  16. Clostridium difficile: A Cause of Diarrhea in Children

    MedlinePLUS

    ADVICE FOR PATIENTS Clostridium difficile : A Cause of Diarrhea in Children C lostridium difficile is a bacterium ... the most common cause of health care–associated diarrhea in the United States. While most health care– ...

  17. The ClosTron: Mutagenesis in Clostridium refined and streamlined

    Microsoft Academic Search

    John T. Heap; Sarah A. Kuehne; Muhammad Ehsaan; Stephen T. Cartman; Clare M. Cooksley; Jamie C. Scott; Nigel P. Minton

    2010-01-01

    The recent development of the ClosTron Group II intron directed mutagenesis tool for Clostridium has advanced genetics in this genus, and here we present several significant improvements. We have shown how marker re-cycling can be used to construct strains with multiple mutations, demonstrated using FLP\\/FRT in Clostridium acetobutylicum; tested the capacity of the system for the delivery of transgenes to

  18. 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

    by immunohistochemical staining for the presence of viral infections including bovine viral diarrhea (BVD), however hrs later) with blood secreted from the rectum and bloody diarrhea was found in the area. Externally

  19. 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...

  20. Novel real-time PCR assay for simultaneous detection and differentiation of Clostridium chauvoei and Clostridium septicum in clostridial myonecrosis.

    PubMed

    Halm, Anna; Wagner, Martin; Köfer, Josef; Hein, Ingeborg

    2010-04-01

    A real-time PCR assay based on the 16S rRNA gene sequence was designed for differentiation of blackleg-causing Clostridium chauvoei and Clostridium septicum, a phylogenetically closely related bacterium responsible for malignant edema. In order to exclude false-negative results, an internal amplification control was included in the assay. A set of three probes, one specific for C. chauvoei, one specific for C. septicum, and one specific for both species, permitted unequivocal detection of C. chauvoei in tests of 32 Clostridium sp. strains and 10 non-Clostridium strains. The assay proved to be sensitive, detecting one genome of C. chauvoei or C. septicum per PCR and 1.79 x 10(3) C. chauvoei cells/g artificially contaminated muscle tissue. In tests of 11 clinical specimens, the real-time PCR assay yielded the same results as an established conventional PCR method. PMID:20129968

  1. 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

  2. Biotechnological potential of Clostridium butyricum bacteria

    PubMed Central

    Szymanowska-Powa?owska, Daria; Orczyk, Dorota; Leja, Katarzyna

    2014-01-01

    In response to demand from industry for microorganisms with auspicious biotechnological potential, a worldwide interest has developed in bacteria and fungi isolation. Microorganisms of interesting metabolic properties include non-pathogenic bacteria of the genus Clostridium, particularly C. acetobutylicum, C. butyricum and C. pasteurianum. A well-known property of C. butyricum is their ability to produce butyric acid, as well as effectively convert glycerol to 1,3-propanediol (38.2 g/L). A conversion rate of 0.66 mol 1,3-propanediol/mol of glycerol has been obtained. Results of the studies described in the present paper broaden our knowledge of characteristic features of C. butyricum specific isolates in terms of their phylogenetic affiliation, fermentation capacity and antibacterial properties. PMID:25477923

  3. Prevalence of Clostridium difficile in horses.

    PubMed

    Medina-Torres, Carlos E; Weese, J Scott; Staempfli, Henry R

    2011-08-26

    Fecal samples were collected to establish the apparent prevalence of Clostridium difficile shedding in Standardbred and Thoroughbred racehorses housed at 4 racetracks and 2 breeding facilities, and in horses admitted to a referral large animal clinic. Forty-one (7.59%) of 540 racetrack horses, seven (5.83%) of 120 breeding farm horses, and four (4.88%) out of 82 horses admitted to the referral clinic were culture-positive for C. difficile. An overall fecal culture prevalence of 7.01% for C. difficile was identified in 742 fecal samples. PCR-ribotyping and toxin gene identification was performed and seventeen 17 PCR-ribotypes were identified among the 52 C. difficile isolates. PMID:21570780

  4. Clostridium difficile associated disease in Thailand.

    PubMed

    Wongwanich, S; Ramsiri, S; Vanasin, B; Khowsaphit, P; Tantipatayangkul, P; Phan-urai, R

    1990-09-01

    Over a twenty-six month period 383 fecal specimens from 269 diarrheal patients and 114 control patients were examined for Clostridium difficile and its cytotoxin. C. difficile was isolated from 13 (4.8%) of overall age group of diarrheal patients and from 3 (2.6%) of controls. Fecal cytotoxin was detected in 106 (52.5%) of 203 diarrheal patients and in 17 (22.4%) of 76 controls. Sixty-one percent of antibiotic-associated diarrheal patients (less than 3 years) and 51% of non-antibiotic associated diarrhea patients had fecal cytotoxin. Enteric pathogens other than C. difficile were detected in 0.7-7.4% of the patients studied. These data suggest that C. difficile associated disease may be frequently encountered in such a developing region studied. Routine diagnosis for C. difficile in diarrheal patients appears to be warranted. PMID:2075479

  5. Transcriptional organization of the Clostridium acetobutylicum genome

    PubMed Central

    Paredes, Carlos J.; Rigoutsos, Isidore; Papoutsakis, E. Terry

    2004-01-01

    Prokaryotic genes are frequently organized in multicistronic operons (or transcriptional units, TUs), and usually the regulatory motifs for the whole TU are located upstream of the first TU gene. Although the number of sequenced genomes has increased dramatically, experimental information on TU organization is extremely limited. Even for organisms as extensively studied as Escherichia coli and Bacillus subtilis, TU annotation is far from complete. It therefore becomes imperative to rely on computational approaches to complement experimental information. Here we present a TU map for the obligate anaerobe Clostridium acetobutylicum ATCC 824. This map is largely based on the distance between pairs of consecutive genes but enhanced and refined by predictions of several types of promoters (?A, ?E and ?F/G) and rho-independent terminator structures. Based on the set of known C.acetobutylicum TUs, the presented TU map offers an 88% prediction accuracy. PMID:15060177

  6. Clostridium septicum Aortitis and Cecal Adenocarcinoma.

    PubMed

    Moseley, Brian; Mwirigi, Nicola W; Bowen, Juan

    2010-01-01

    Clostridium septicum aortitis is a rare infection that is highly associated with underlying malignancy. We present a case of an 82-year-old male diagnosed with both C. septicum aortitis and a high-grade cecal tubulovillous adenoma. The patient was offered aortic resection; however, he opted for only suppressive antibiotic therapy and a right hemicolectomy with ileocolonic anastomosis. He ultimately passed away 75 days following admission. The authors report on the connection between C. septicum aortitis and malignancy. The authors also discuss the need for prompt treatment with antibiotics once the infection is identified and the consideration of aortic resection given the risk of aneurysmal change with aortic dissection or rupture. PMID:20224641

  7. Biotechnological potential of Clostridium butyricum bacteria.

    PubMed

    Szymanowska-Powa?owska, Daria; Orczyk, Dorota; Leja, Katarzyna

    2014-01-01

    In response to demand from industry for microorganisms with auspicious biotechnological potential, a worldwide interest has developed in bacteria and fungi isolation. Microorganisms of interesting metabolic properties include non-pathogenic bacteria of the genus Clostridium, particularly C. acetobutylicum, C. butyricum and C. pasteurianum. A well-known property of C. butyricum is their ability to produce butyric acid, as well as effectively convert glycerol to 1,3-propanediol (38.2 g/L). A conversion rate of 0.66 mol 1,3-propanediol/mol of glycerol has been obtained. Results of the studies described in the present paper broaden our knowledge of characteristic features of C. butyricum specific isolates in terms of their phylogenetic affiliation, fermentation capacity and antibacterial properties. PMID:25477923

  8. First isolation of Clostridium amygdalinum from a patient with chronic osteitis.

    PubMed

    Carlier, Jean-Philippe; Manich, Maria; Loïez, Caroline; Migaud, Henri; Courcol, René J

    2006-10-01

    We describe a case of osteitis caused by a new and unusual Clostridium species, Clostridium amygdalinum, an environmental, moderately thermophilic bacterium. This is the first documented report of human infection caused by this organism. PMID:17021125

  9. Cloning, developmental regulation and neural localization of rat epsilon-sarcoglycan.

    PubMed

    Xiao, Jianfeng; LeDoux, Mark S

    2003-11-26

    Mutations in the gene for epsilon sarcoglycan (epsilon-SG) are associated with a disorder of the central nervous system, the myoclonus-dystonia syndrome (MDS; DYT11). In contrast, mutations of other sarcoglycan family members lead to limb-girdle muscular dystrophies. To establish the framework for functional studies of epsilon-SG, we cloned rat epsilon-SG cDNA, quantified epsilon-SG mRNA levels in neural and non-neural tissues at different developmental time points with relative quantitative multiplex real-time reverse transcriptase PCR (RT-PCR), and characterized the distribution of epsilon-SG mRNA in brain with in situ hybridization. Rat epsilon-SG cDNA contains an open reading frame (ORF) of 1311 bp that encodes a 437-amino acid (aa) protein with 95.9% and 98.2% identity to human and mouse epsilon-SG amino acid sequences, respectively. Using real-time RT-PCR, epsilon-SG was detected in both neural (cerebellar cortex, striatum, cerebral cortex, thalamus, hippocampus) and non-neural (muscle, liver, kidney, heart) tissues at each developmental time point tested [Embryonic Day 20 (E20), Postnatal Day 1 (P1), P7, P14, P36, 6 months, 1.5 years). Levels of epsilon-SG mRNA were highest at E20 in all tissues. The developmental regulation of epsilon-SG mRNA expression was most striking in muscle with E20 and early postnatal epsilon-SG mRNA levels over 10 times higher than those seen in adult rats. In adult rats, epsilon-SG mRNA levels were several-fold higher in brain, particularly cerebellar cortex, than in muscle. Radioactive in situ hybridization showed that epsilon-SG mRNA was widely distributed in rat brain. Robust hybridization signal was obtained from regions with dense neuronal packing such as the hippocampus, cerebellar molecular layer, and cerebral cortex. Our results suggest that epsilon-SG participates in the development of both neural and non-neural tissues and contributes to neuronal structure in the adult central nervous system. PMID:14625080

  10. Crucial role of phospholipase C epsilon in skin inflammation induced by tumor-promoting phorbol ester.

    PubMed

    Ikuta, Shuzo; Edamatsu, Hironori; Li, Mingzhen; Hu, Lizhi; Kataoka, Tohru

    2008-01-01

    In two-stage skin chemical carcinogenesis, phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) acts as a promoter essential for clonal expansion of the initiated cells carrying the activated ras oncogenes. Although protein kinase C (PKC) isozymes are the main targets of TPA, their role in tumor promotion remains controversial. We previously reported that mice lacking a Ras/Rap effector phospholipase C epsilon (PLC epsilon(-/-) mice) exhibited marked resistance to tumor formation in the two-stage skin carcinogenesis. PLC epsilon(-/-) mice also failed to exhibit basal layer cell proliferation and epidermal hyperplasia induced by TPA, suggesting a role of PLC epsilon in tumor promotion. Here, we show that PLC epsilon(-/-) mice exhibit resistance to TPA-induced skin inflammation as assessed by reduction in edema, granulocyte infiltration, and expression of a proinflammatory cytokine, interleukin-1 alpha (IL-1 alpha). On the other hand, the proliferative potentials of keratinocytes or dermal fibroblasts in culture remain unaffected by the PLC epsilon background, suggesting that the PLC epsilon's role in tumor promotion may be ascribed to augmentation of inflammatory responses. In dermal fibroblast primary culture, TPA can induce activation of the PLC epsilon lipase activity, which leads to the induction of IL-1 alpha expression. Experiments using small interfering RNA-mediated knockdown indicate that this activation is mediated by Rap1, which is activated by a TPA-responsive guanine nucleotide exchange factor RasGRP3. Moreover, TPA-induced activation of Rap1 and PLC epsilon is inhibited by a PKC inhibitor GF109203X, indicating a crucial role of PKC in signaling from TPA to PLC epsilon. These results imply that two TPA targets, RasGRP3 and PKC, are involved in TPA-induced inflammation through PLC epsilon activation, leading to tumor promotion. PMID:18172297

  11. 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.

  12. Effect of magnetic field on TTT diagram of successive ??epsilon'??' martensitic transformation in SUS304L stainless steel

    NASA Astrophysics Data System (ADS)

    Lee, J. H.; Fukuda, T.; Kakeshita, T.

    2009-03-01

    We have investigated the effect of magnetic field on TTT diagram of successive ??epsilon'??' martensitic transformation in a solution-treated SUS304L stainless steel in order to clarify kinetics of the successive martensitic transformation. The TTT diagram of successive ??epsilon'??' martensitic transformation shows a C-curve with a nose temperature located at 103 K. Incubation time of the successive ??epsilon'??' martensitic transformation shortens with increasing the strength of magnetic field. However, the nose temperature does not change significantly under the magnetic field. An in-situ optical micrograph suggests that the successive ??epsilon'??' transformation probably proceeds by isothermal ??epsilon' martensitic transformation followed by athermal epsilon'??' martensitic transformation.

  13. Unitary Fermi gas, epsilon expansion, and nonrelativistic conformal field theories

    E-print Network

    Yusuke Nishida; Dam Thanh Son

    2010-04-20

    We review theoretical aspects of unitary Fermi gas (UFG), which has been realized in ultracold atom experiments. We first introduce the epsilon expansion technique based on a systematic expansion in terms of the dimensionality of space. We apply this technique to compute the thermodynamic quantities, the quasiparticle spectrum, and the critical temperature of UFG. We then discuss consequences of the scale and conformal invariance of UFG. We prove a correspondence between primary operators in nonrelativistic conformal field theories and energy eigenstates in a harmonic potential. We use this correspondence to compute energies of fermions at unitarity in a harmonic potential. The scale and conformal invariance together with the general coordinate invariance constrains the properties of UFG. We show the vanishing bulk viscosities of UFG and derive the low-energy effective Lagrangian for the superfluid UFG. Finally we propose other systems exhibiting the nonrelativistic scaling and conformal symmetries that can be in principle realized in ultracold atom experiments.

  14. Gearing Up for Epsilon Aurigae's First Eclipse of the Millennium

    E-print Network

    J. L. Hopkins; L. Schanne; R. E. Stencel

    2008-07-17

    The mysterious 3rd magnitude long period eclipsing binary star system epsilon Aurigae is predicted to be starting its 2 year eclipse in the late summer of 2009. While this is when the real excitement starts, much is to be learned before first contact. This paper discusses current observational results that have accumulated thus far, using photometric monitoring, H-alpha spectroscopy and with other data sources. Key among the findings are that (1) the low amplitude light variation quasi-period has decreased significantly over the past 20 years, and (2) that the duration of egress, eclipse-to-eclipse has been decreasing, while the duration of total eclipse has been increasing. The website for the observing campaign is: www.du.edu/~rstencel/epsaur.htm .

  15. 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

  16. 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

  17. 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...

  18. 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.

  19. Ferromagnetic resonance in $\\epsilon$-Co magnetic composites

    E-print Network

    Chalapat, Khattiya; Huuppola, Maija; Koponen, Lari; Johans, Christoffer; Ras, Robin H A; Ikkala, Olli; Oksanen, Markku A; Seppälä, Eira; Paraoanu, G S

    2014-01-01

    We investigate the electromagnetic properties of assemblies of nanoscale $\\epsilon$-cobalt crystals with size range between 5 nm to 35 nm, embedded in a polystyrene (PS) matrix, at microwave (1-12 GHz) frequencies. We investigate the samples by transmission electron microscopy (TEM) imaging, demonstrating that the particles aggregate and form chains and clusters. By using a broadband coaxial-line method, we extract the magnetic permeability in the frequency range from 1 to 12 GHz, and we study the shift of the ferromagnetic resonance with respect to an externally applied magnetic field. We find that the zero-magnetic field ferromagnetic resonant peak shifts towards higher frequencies at finite magnetic fields, and the magnitude of complex permeability is reduced. At fields larger than 2.5 kOe the resonant frequency changes linearly with the applied magnetic field, demonstrating the transition to a state in which the nanoparticles become dynamically decoupled. In this regime, the particles inside clusters can ...

  20. 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.

  1. Shock initiation of an {epsilon}-CL-20-estane formulation

    SciTech Connect

    Tarver, C.M.; Simpson, R.L.; Urtiew, P.A.

    1995-07-19

    The shock sensitivity of a pressed solid explosive formulation, LX-19, containing 95.2% by weight epsilon phase 2,4,6,8,10,12-hexanitrohexaazaisowurtzitane (HNIW) and 4.8% 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% HMX and 4.5% 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 model for LX-19 was developed which differed from those for HMX-inert binder formulations only by a 25% higher hot spot growth rate.

  2. Campaign Photometry During The 2010 Eclipse Of Epsilon Aurigae

    NASA Astrophysics Data System (ADS)

    Hopkins, Jeff; Stencel, R. E.

    2011-01-01

    Epsilon Aurigae is a long period (27.1 years) eclipsing binary star system with an eclipse that lasts nearly 2 years, but with severe ambiguities about component masses and shape. The current eclipse began on schedule in August of 2009. During the previous, 1982-1984 eclipse, an International Campaign was formed to coordinate a detailed study of the system. While that Campaign was deemed successful, the evolutionary status of the star system remained unclear. Epsilon Aurigae has been observed nearly continuously since the 1982 eclipse. The current Campaign was officially started in 2006. In addition to a Yahoo forum we have a dedicated web site and more than 18 online newsletters reporting photometry, spectroscopy, interferometry and polarimetry data. High quality UBVRIJH band photometric data since before the start of the current eclipse has been submitted. We explore the color differences among the light curves in terms of eclipse phases and archival data. At least one new model of the star system has been proposed since the current Campaign began: a low mass but very high luminosity F star plus a B star surrounded by a debris disk. The current eclipse and in particular the interferometry and spectroscopic data have caused new thoughts on defining eclipsing variable star contact points and phases of an eclipse. Second contact may not be the same point as start of totality and third contact may not be the same point as the start of egress and end of totality. In addition, the much awaited mid-eclipse brightening may or may not have appeared. This paper identifies the current Campaign contributors and the photometric data. This work was supported in part by the bequest of William Herschel Womble in support of astronomy at the University of Denver, by NSF grant 1016678 to the University of Denver.

  3. Constraints from Asymmetric Heating: Investigating the Epsilon Aurigae Disk

    NASA Astrophysics Data System (ADS)

    Pearson, Richard L., III; Stencel, Robert E.

    2015-01-01

    Epsilon Aurigae is a long-period eclipsing binary that likely contains an F0Ia star and a circumstellar disk enshrouding a hidden companion, assumed to be a main-sequence B star. High uncertainty in its parallax has kept the evolutionary status of the system in question and, hence, the true nature of each component. This unknown, as well as the absence of solid state spectral features in the infrared, requires an investigation of a wide parameter space by means of both analytic and Monte Carlo radiative transfer (MCRT) methods. The first MCRT models of epsilon Aurigae that include all three system components are presented here. We seek additional system parameter constraints by melding analytic approximations with MCRT outputs (e.g., dust temperatures) on a first-order level. The MCRT models investigate the effects of various parameters on the disk-edge temperatures; these include two distances, three particle size distributions, three compositions, and two disk masses, resulting in 36 independent models. Specifically, the MCRT temperatures permit analytic calculations of effective heating and cooling curves along the disk edge. These are used to calculate representative observed fluxes and corresponding temperatures. This novel application of thermal properties provides the basis for utilization of other binary systems containing disks. We find degeneracies in the model fits for the various parameter sets. However, the results show a preference for a carbon disk with particle size distributions >=10 ?m. Additionally, a linear correlation between the MCRT noon and basal temperatures serves as a tool for effectively eliminating portions of the parameter space.

  4. Clostridium chromiireducens sp. nov., isolated from Cr(VI)-contaminated soil

    E-print Network

    Florida, University of

    with group I of the genus Clostridium. Of strains within this cluster, strain GCAF-1T shared the highest 16S, strain GCAF-1T (5DSM 23318T 5KCTC 5935T ) represents a novel species of the genus Clostridium, for which widely studied. The genus Clostridium forms one of the largest genera of Gram-positive bacteria and its

  5. Distribution of Clostridial cry Like Genes Among Bacillus thuringiensis and Clostridium Strains

    Microsoft Academic Search

    Frédérique Barloy; Marguerite-M. Lecadet; Armelle Delécluse

    1998-01-01

    .   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

  6. Occurrence of Clostridium tyrobutyricum in cattle slurry and fresh forage grasses

    Microsoft Academic Search

    Zsuzsanna Langó; Helvi Heinonen-Tanski

    1995-01-01

    A comparative, numerical, taxonomic analysis was carried out on 24 selected strains of Clostridium spp. isolated from cattle slurries and fresh forage grasses fertilized with slurries, as well as on 10 culture-collection Clostridium tyrobutyricum strains for reference. The results showed an unexpectedly low occurrence of the Clostridium tyrobutyricum, with only one of the 24 strains isolated from slurry belonging to

  7. Ultrafast plasmonics using transparent conductive oxide hybrids in the epsilon-near-zero regime

    E-print Network

    Zheludev, Nikolay

    Ultrafast plasmonics using transparent conductive oxide hybrids in the epsilon-near-zero regime plasmonics using transparent conductive oxide hybrids in the epsilon-near-zero regime Daniel Traviss,1 Roman March 2013) The dielectric response of transparent conductive oxides near the bulk plasmon frequency

  8. Low-energy density correlation function of a degenerate unitary Fermi gas from the {epsilon} expansion

    SciTech Connect

    Kryjevski, Andrei [Department of Physics, North Dakota State University, Fargo, North Dakota 58108 (United States)

    2010-04-15

    Using the {epsilon} expansion proposed in Y. Nishida and D. T. Son [Phys. Rev. Lett. 97, 050403 (2006)], we calculate the density correlation function and the dynamic structure function of the unitary Fermi gas at zero temperature to next-to-leading order in {epsilon} for excitation energies below the quasiparticle threshold.

  9. epsilon-N-trimethyllysine availability regulates the rate of carnitine biosynthesis in the growing rat

    SciTech Connect

    Rebouche, C.J.; Lehman, L.J.; Olson, L.

    1986-05-01

    Rates of carnitine biosynthesis in mammals depend on the availability of substrates and the activity of enzymes subserving the pathway. This study was undertaken to test the hypothesis that the availability of epsilon-N-trimethyllysine is rate-limiting for synthesis of carnitine in the growing rat and to evaluate diet as a source of this precursor for carnitine biosynthesis. Rats apparently absorbed greater than 90% of a tracer dose of (methyl-/sup 3/H)epsilon-N-trimethyllysine, and approximately 30% of that was incorporated into tissues as (/sup 3/H)carnitine. Rats given oral supplements of epsilon-N-trimethyllysine (0.5-20 mg/d), but no dietary carnitine, excreted more carnitine than control animals receiving no dietary epsilon-N-trimethyllysine or carnitine. Rates of carnitine excretion increased in a dose-dependent manner. Tissue and serum levels of carnitine also increased with dietary epsilon-N-trimethyllysine supplementation. There was no evidence that the capacity for carnitine biosynthesis was saturated even at the highest level of oral epsilon-N-trimethyllysine supplementation. Common dietary proteins (casein, soy protein and wheat gluten) were found to be poor sources of epsilon-N-trimethyllysine for carnitine biosynthesis. The results of this study indicate that the availability of epsilon-N-trimethyllysine limits the rate of carnitine biosynthesis in the growing rat.

  10. The extent of mitochondrial F1-ATPase and adenine nucleotide carrier activity with epsilon-ATP.

    PubMed

    Kaplan, R S; Coleman, P S

    1978-02-01

    1. The use of 1,N6-ethenoadenosine 5'-triphosphate (epsilon-ATP), a synthetic, fluorescent analog of ATP, by whole rat liver mitochondria and by submitochondrial particles produced via sonication has been studied. 2. Direct [3H]adenine nucleotide uptake studies with isolated mitochondria, indicate the epsilon-[3H]ATP is not transported through the inner membrane by the adenine nucleotide carrier and is therefore not utilized by the 2,4-dinitrophenol-sensitive F1-ATPase (EC 3.6.1.3) that functions in oxidative phosphorylation. However, epsilon-ATP is hydrolyzed by a Mg2+-dependent, 2,4-dinitrophenol-insensitive ATPase that is characteristic of damaged mitochondria. 3. epsilon-ATP can be utilized quite well by the exposed F1-ATPase of sonic submitochondrial particles. This epsilon-ATP hydrolysis activity is inhibited by oligomycin and stimulated by 2,4-dinitrophenol. The particle F1-ATPase displays similar Km values for both ATP and epsilon-ATP; however, the V with ATP is approximately six times greater than with epsilon-ATP. 4. Since epsilon-ATP is a capable substrate for the submitochondrial particle F1-ATPase, it is proposed that the fluorescent properties of this ATP analog might be employed to study the submitochondrial particle F1-ATPase complex, and its response to various modifiers of oxidative phosphorylation. PMID:145875

  11. 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).

  12. Clostridium bolteae sp. nov., isolated from human sources.

    PubMed

    Song, Yuli; Liu, Chengxu; Molitoris, Denise R; Tomzynski, Thomas J; Lawson, Paul A; Collins, Matthew D; Finegold, Sydney M

    2003-03-01

    Seven obligately anaerobic, gram-positive, rod-shaped, spore-forming organisms isolated from human sources were characterized using phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing showed that the strains were genetically highly related to each other (displaying >99% sequence similarity) and represent a previously unknown sub-line within the Clostridium coccoides rRNA group of organisms. Strains of the unidentified bacterium used carbohydrate as fermentable substrates, producing acetic acid and lactic acid as the major products of glucose metabolism. The closest described species to the novel bacterium corresponded to Clostridium clostridioforme, although a 16S rRNA sequence divergence of 3% demonstrated they represent different species. Genomic DNA-DNA pairing studies confirmed the separateness of the unknown species and Clostridium clostridioforme. Based on phenotypic and phylogenetic evidence, it is therefore proposed that the unknown bacterium, be classified as Clostridium bolteae sp. nov. The type strain of Clostridium bolteae is WAL 16351T (= ATCC(T) = BAA-613T, CCUG(T) = 46953T). PMID:12747414

  13. Polymerase Epsilon Mutations Accelerate Mutation Rates in Colorectal and Endometrial Cancer - David Wheeler, TCGA Scientific Symposium 2012

    Cancer.gov

    Home News and Events Multimedia Library Videos Polymerase Epsilon Mutations in Colorectal and Endometrial Cancer - David Wheeler Polymerase Epsilon Mutations Accelerate Mutation Rates in Colorectal and Endometrial Cancer - David Wheeler, TCGA Scientific

  14. 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

  15. Progesterone Analogs Influence Germination of Clostridium sordellii and Clostridium difficile Spores In Vitro?

    PubMed Central

    Liggins, Marc; Ramirez, Norma; Magnuson, Natiera; Abel-Santos, Ernesto

    2011-01-01

    Clostridium sordellii and Clostridium difficile are closely related anaerobic Gram-positive, spore-forming human pathogens. C. sordellii and C. difficile form spores that are believed to be the infectious form of these bacteria. These spores return to toxin-producing vegetative cells upon binding to small molecule germinants. The endogenous compounds that regulate clostridial spore germination are not fully understood. While C. sordellii spores require three structurally distinct amino acids to germinate, the occurrence of postpregnancy C. sordellii infections suggests that steroidal sex hormones might regulate its capacity to germinate. On the other hand, C. difficile spores require taurocholate (a bile salt) and glycine (an amino acid) to germinate. Bile salts and steroid hormones are biosynthesized from cholesterol, suggesting that the common sterane structure can affect the germination of both C. sordellii and C. difficile spores. Therefore, we tested the effect of sterane compounds on C. sordellii and C. difficile spore germination. Our results show that both steroid hormones and bile salts are able to increase C. sordellii spore germination rates. In contrast, a subset of steroid hormones acted as competitive inhibitors of C. difficile spore germination. Thus, even though C. sordellii and C. difficile are phylogenetically related, the two species' spores respond differently to steroidal compounds. PMID:21478359

  16. 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.

  17. A prediction model for Clostridium difficile recurrence

    PubMed Central

    LaBarbera, Francis D.; Nikiforov, Ivan; Parvathenani, Arvin; Pramil, Varsha; Gorrepati, Subhash

    2015-01-01

    Background Clostridium difficile infection (CDI) is a growing problem in the community and hospital setting. Its incidence has been on the rise over the past two decades, and it is quickly becoming a major concern for the health care system. High rate of recurrence is one of the major hurdles in the successful treatment of C. difficile infection. There have been few studies that have looked at patterns of recurrence. The studies currently available have shown a number of risk factors associated with C. difficile recurrence (CDR); however, there is little consensus on the impact of most of the identified risk factors. Methods Our study was a retrospective chart review of 198 patients diagnosed with CDI via Polymerase Chain Reaction (PCR) from January 2009 to Jun 2013. In our study, we decided to use a machine learning algorithm called the Random Forest (RF) to analyze all of the factors proposed to be associated with CDR. This model is capable of making predictions based on a large number of variables, and has outperformed numerous other models and statistical methods. Results We came up with a model that was able to accurately predict the CDR with a sensitivity of 83.3%, specificity of 63.1%, and area under curve of 82.6%. Like other similar studies that have used the RF model, we also had very impressive results. Conclusions We hope that in the future, machine learning algorithms, such as the RF, will see a wider application. PMID:25656667

  18. Parameters affecting solvent production by Clostridium pasteurianum

    SciTech Connect

    Dabrock, B.; Bahl, H.; Gottschalk, G. (Georg-August-Univ. Goettingen (Germany))

    1992-04-01

    The effect of pH, growth rate, phosphate and iron limitation, carbon monoxide, and carbon source on product formation by Clostridium pasteurianum was determined. Under phosphate limitation, glucose was fermented almost exclusively to acetate and butyrate independently of the pH and growth rate. Iron limitation caused lactate production (38 mol/100 mol) from glucose in batch and continuous culture. At 15% (vol/vol) carbon monoxide in the atmosphere, glucose was fermented to ethanol (24 mol/100 mol), lactate (32 mol/100 mol), and butanol (36 mol/100 mol) in addition to the usual products, acetate (38 mol/100 mol) and butyrate (17 mol/100 mol). During glycerol fermentation, a completely different product pattern was found. In continuous culture under phosphate limitation, acetate and butyrate were produced only in trace amounts, whereas ethanol (30 mol/10 mol), butanol (18 mol/100 mol), and 1,3-propanediol (18 mol/100 mol) were the major products. Under iron limitation, the ratio of these products could be changed in favor of 1,3-propanediol (34 mol/100 mol). In addition, lactate was produced in significant amounts (25 mol/100 mol). The tolerance of C. pasteurianum to glycerol was remarkably high; growth was not inhibited by glycerol concentrations up to 17% (wt/vol). Increasing glycerol concentrations favored the production of 1,3-propanediol.

  19. Effects of butanol on Clostridium acetobutylicum

    SciTech Connect

    Bowles, L.K.; Ellefson, W.L.

    1985-11-01

    The internal pH of Clostridium acetobutylicum was determined at various stages during the growth of the organism. Even in the presence of significant quantities of acetic, butyric, and lactic acids, an internal pH of 6.2 was maintained. Experiments using N,N'-dicyclohexylcarbodiimide indicated that a functioning H/sup +/-ATPase is necessary for internal pH control. Butanol, one of the end products of the fermentation, had numerous harmful effects on C. acetobutylicum. At a concentration high enough to inhibit growth, butanol destroyed the ability of the cell to maintain internal pH, lowered the intracellular level of ATP, and inhibited glucose uptake. Experiments done at two different external pH values suggested that the butanol-mediated decrease in ATP concentration was independent of the drop in internal pH. Glucose uptake was not affected by arsenate, suggesting that uptake was not ATP dependent. The effects of butanol on C. acetobutylicum are complex, inhibiting several interrelated membrane processes.

  20. Clostridium difficile infection in adult hamsters.

    PubMed

    Chang, J; Rohwer, R G

    1991-12-01

    Diarrhea was encountered in a group of adult female golden Syrian hamsters (Mesocricetus auratus) used for titrating the scrapie agent. Ninety percent of the cases occurred in animals over 210 days old even though animals of all age groups lived in the colony concurrently. The cause of diarrhea was investigated in both uninoculated animals and those receiving greater than a limiting dilution of scrapie infectivity, i.e., animals that were not expected to contract the experimental scrapie disease. Three forms of diarrhea were observed. The most commonly encountered was profuse and watery. A chronic form presented with semiformed, thin fecal material smearing the retroperitoneal region. Hemorrhagic diarrhea was observed rarely. Mortality was high among animals with acute watery or hemorrhagic diarrhea. Animals with semiformed soft stools were dehydrated, had a roughened hair-coat, and hunched back. Cardinal lesions were necrosis, inflammation, and mucosal hyperplasia of the cecum and colon and cholangiohepatitis with amyloid deposition. Diffuse renal amyloidosis was present in chronic cases. Toxigenic, cytotoxin B-positive Clostridium difficile was isolated from a majority of affected animals. Cytotoxin B was also present in cecal homogenates of diarrheic animals with C. difficile. The pathological and microbiologic findings indicated a typhlitis and colitis in adult hamsters that was associated with C. difficile infection. PMID:1667195

  1. Diagnosis and management of Clostridium difficile infection.

    PubMed

    Korman, Tony M

    2015-02-01

    There have been dramatic changes in the epidemiology of Clostridium difficile infection (CDI), with increases in incidence and severity of disease, attributed to the emergence of a fluoroquinolone-resistant "hypervirulent" strain, ribotype 027. C. difficile is now the most common pathogen causing hospital-acquired infection in U.S. hospitals, and community-acquired infections are increasing. The diagnosis of CDI is based on a combination of signs and symptoms, confirmed by laboratory tests. Clinical manifestations of CDI can range from asymptomatic colonization to severe pseudomembranous colitis and death. Many aspects of laboratory diagnosis of CDI remain contentious. Toxin enzyme immunoassays are too insensitive to be used alone, while nucleic acid amplification tests have emerged as an option, either as a stand-alone test or as part of a multitest algorithm. Oral vancomycin and metronidazole have been the recommended antimicrobial therapy options, and fidaxomicin is an effective new alternative. There is ongoing concern regarding the potential inferiority of metronidazole, in particular for severe CDI. Management of severe CDI and recurrent CDI continue to represent major treatment challenges. Biological therapies for the restoration of the intestinal microbiota (e.g., fecal microbiota transplantation) and monoclonal antibody therapy are promising approaches for CDI management, in particular troublesome recurrent CDI. This review will concentrate on the diagnosis and management of CDI in adults. PMID:25643269

  2. Clostridium difficile infection and inflammatory bowel disease.

    PubMed

    Cojocariu, Camelia; Stanciu, Carol; Stoica, Oana; Singeap, Ana Maria; Sfarti, Catalin; Girleanu, Irina; Trifan, Anca

    2014-12-01

    Over the past 15 years, Clostridium difficile infection (CDI) in patients with inflammatory bowel disease (IBD) has increased both in incidence and severity. Traditional risk factors for CDI are similar in IBD and non-IBD populations, but there is a significant proportion of IBD patients which have distinctive characteristics. Patients with ulcerative colitis (UC) are more susceptible to CDI and have more severe outcomes than those with Crohn's disease (CD). CDI may be difficult to distinguish from an IBD flare due to similar clinical presentation, and therefore screening for CDI is recommended at every flare in such patients. Several studies showed worse clinical outcomes in IBD patients with CDI, including longer hospital stay, higher colectomy and mortality rates than in those without CDI. Vancomycin and metronidazole appear to have similar efficacy in patients with moderate disease, but vancomycin is preferred in severe disease. Measures must be taken to prevent the spread of infection. Clinicians should have a high index of suspicion for CDI when evaluating a patient with IBD flare, as rapid detection and prompt treatment of infection improve outcomes. This review summarizes the available literature on epidemiology, risk factors, clinical aspects, diagnostic methods, treatment, outcome, and prevention of CDI in IBD patients. PMID:25599768

  3. [Optimization of culture conditions for Clostridium cellulolyticum].

    PubMed

    Wang, Lang; Liu, Zhidan; Wang, Tianmin; Wu, Xiao; Zhang, Chong; Wang, Qunhui; Xing, Xinhui

    2013-03-01

    Clostridium cellulolyticum, as one of obligate anaerobic bacteria capable of secreting cellulosome, has not been efficiently cultured due to its strict requirement of growing conditions. In this study, culture conditions of C. cellulolyticum were optimized using response surface methodology. Plackett-Burman design was first used to screen the dominant impact factors for the growth of C. cellulolyticum, which were determined as yeast extract concentration, cellobiose concentration and culture temperature. The steepest ascent path design was then applied to gain the suitable range close to the optimal culture conditions for obtaining high cell density. The central composite design and the response surface analysis were finally used to determine the optimal levels of the influential factors, which were 3 g/L for yeast extract concentration, 7 g/L cellobiose concentration and 34 degrees C for culture temperature. The optimized medium was used for flask culture, and OD600 of C. cellulolyticum was increased from 0.303 to 0.586. With a pH-controlled fermentor at batch mode, OD600 reached 3.432, which was 2.8 times higher than elsewhere reported. These results support further study on the high-density culture of C. cellulolyticum and its application. PMID:23789280

  4. Clostridium septicum sepsis and its implications.

    PubMed

    Khalid, Muhammad; Lazarus, Rajeka; Bowler, Ian C J W; Darby, Chris

    2012-01-01

    An elderly gentleman, who had 12 years earlier been successfully treated for colon cancer, presented with fever, rigours, right upper quadrant abdominal pain and tenderness. A CT of the abdomen revealed a colonic mass distal to the hepatic flexure with multiple gas locules and a walled off perforation. He underwent a right hemicolectomy. Histology confirmed multifocal colonic adenocarcinoma. His admission blood cultures grew Clostridium septicum. A week postoperatively he developed intermittent fevers and abdominal pain. Repeat CT revealed an abdominal collection adjacent to the new anastomosis, but more importantly, a sharply shouldered aneurysmal dilation of the infra-renal abdominal aorta. These findings prompted immediate surgical drainage of the collection, repair of the anastomostic leak, resection of the infected aortic aneurysm and replacement with a tube graft. This case highlights the clinical significance of C septicum bacteraemia: its association with occult colonic malignancy and with mycotic aneurysm formation. Clostridia isolated from blood cultures should not be dismissed as contaminants but fully identified to ensure appropriate patient management. PMID:22962388

  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. Review article Clostridial diseases of small ruminants

    E-print Network

    Paris-Sud XI, Université de

    ) Abstract - Members of the genus Clo.stridium are extraordinarily diverse in their natural habi- tats, and Clostridium sont extraordinairement divers dans leur habitat naturel, et, lorsqu'ils ont pénétré chez leur INFECTIONS 2.1. Clostridium perfringens Clostridium perf'ringen.s is the most important cause of clostridial

  7. The small subunits of human and mouse DNA polymerase epsilon are homologous to the second largest subunit of the yeast Saccharomyces cerevisiae DNA polymerase epsilon.

    PubMed Central

    Jokela, M; Mäkiniemi, M; Lehtonen, S; Szpirer, C; Hellman, U; Syväoja, J E

    1998-01-01

    Human DNA polymerase epsilon is composed of a 261 kDa catalytic polypeptide and a 55 kDa small subunit of unknown function. cDNAs encoding the small subunit of human and mouse DNA polymerase epsilon were cloned. The predicted polypeptides have molecular masses of 59.469 and 59.319 kDa respectively and they are 90% identical. The human and mouse polypeptides show 22% identity with the 80 kDa subunit of the five subunit DNA polymerase epsilon from the yeast Saccharomyces cerevisiae. The high degree of conservation suggests that the 55 kDa subunit shares an essential function with the yeast 80 kDa subunit, which was earlier suggested to be involved in S phase cell cycle control in a pathway that is able to sense and signal incomplete replication. The small subunits of human and mouse DNA polymerase epsilon also show homology to the C-terminal domain of the second largest subunit of DNA polymerase alpha. The gene for the small subunit of human DNA polymerase epsilon (POLE2) was localized to chromosome 14q21-q22 by fluorescence in situ hybridization. PMID:9443964

  8. 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

  9. 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.

  10. Polarimetry of Epsilon Aurigae from Mid Eclipse to Third Contact

    NASA Astrophysics Data System (ADS)

    Cole, Gary M.; Stencel, Robert E.

    2011-05-01

    In a previous paper, the author discussed the construction of an automated dual beam imaging polarimeter and of observations made in the November 2009 to February 2010 period. Here, we discuss observations and instrumental improvements that span the period from late August 2010 through third and into fourth contacts in Spring 2011. Approximately 930 linear polarization measurements of the target star in BVR bands were obtained during 99 nights of observation. Additional measurements were made of both known polarization standards and zero polarization stars to verify instrument calibration. The polarization of Epsilon Aurigae was observed to vary by nearly 0.4% peak to valley during this period. These variations occurred in several major cycles of varying duration. Measurement error is estimated to be on the order of +/-0.05%. The observed variations resemble excess polarization seen during the 1984 eclipse egress, but may show some differences in detail. During this project, a new optical rotator was developed in conjunction with Optec, Inc., and used for the last two months of observations. This project was initiated at the suggestion of Dr. Robert Stencel at the May 2009 SAS meeting to extend measurements done during the 1984 eclipse by Dr. Jack Kemp and followed up by his student, Dr. Gary Henson thereafter.

  11. The Actual Mass of the Object Orbiting Epsilon Eridani

    NASA Astrophysics Data System (ADS)

    Gatewood, G.

    2000-10-01

    We have tested our 112 Multichannel Astrometric Photometer (MAP) (Gatewood 1987, AJ 94, 213) observations (beginning in 1988) of Epsilon Eridani against the orbital elements provided to us by W. Cochran (private communication). The reduction algorithm is detailed most recently by Gatewood, Han, and Black (2000 ApJ Letters, in press). The seven year period is clearly shown in a variance vs trial periods plot. Although it is near the limit of the current instrument, the astrometric orbital motion is apparent in the residuals to a standard derivation of the star's proper motion and parallax. The astrometric orbital parameters derived by forcing the spectroscopic elements are: semimajor axis = 1.51 +/- 0.44 mas, node of the orbit on the sky = 120 +/- 28 deg, inclination out of the plane of the sky = 46 +/- 17 deg, actual mass = 1.2 +/- 0.33 times that of Jupiter. Our study confirms this object (this is not a minimum mass) as the nearest extrasolar Jupiter mass companion to our solar system. In view of its large orbital eccentricity, however, its exact nature remains unclear.

  12. Small Telescope Infrared Photometry of the epsilon Aurigae Eclipse

    NASA Astrophysics Data System (ADS)

    Rutherford, T. P.

    2012-06-01

    Near-infrared photometry of epsilon Aurigae, in the H- and J-bands, was undertaken during the 2009-2011 eclipse using telescopes of moderate size (8-inch and 14-inch diameter). Instruments of this size successfully collected scientific data in the H- and J-bands. Observations were made from the campus of East Tennessee State University (ETSU), Johnson City, Tennessee, the campus of King College, Bristol, Tennessee, and from the author's home. Signal/Noise ratios of approximately 45 were obtained during times of maximum eclipse. Higher S/N ratios could have been obtained by extending the length of time on target. S/N ratios of almost 100 were obtained outside of eclipse. The infrared light curves produced closely parallel the light curve in the visual range (V), being about 0.5 magnitude brighter in H and 0.7 magnitude brighter in J. The eclipse was easily detected and followed throughout its duration. The rate of ingress was shallower than the rate of egress in both the H- and J-bands. The background variations of the primary star were readily detected.

  13. Citizen Sky, Solving the Mystery of epsilon Aurigae

    NASA Astrophysics Data System (ADS)

    Turner, Rebecca; Price, A.; Kloppenborg, B.; Henden, A.

    2010-01-01

    Citizen Sky is a multi-year, NSF funded citizen science project involving the bright star eps Aur. The project was conceived by the IYA 2009 working group on Research Experiences for Students, Teachers, and Citizen-Scientists. Citizen Sky goes beyond simple observing to include a major data analysis component. The goal is to introduce the participant to the full scientific process from background research to paper writing for a peer-reviewed journal. It begins with a 10 Star Training Program of several types of binary and transient variable stars that are easy to observe from suburban locations with the naked eye. Participants then move on to monitoring the rare and mysterious 2009-2011 eclipse (already underway) of epsilon Aurigae. This object undergoes eclipses only every 27.1 years and each eclipse lasts nearly two years. The star is bright enough to be seen with the naked eye from most urban areas. Training will be provided in observing techniques as well as basic data analysis of photometric and visual datasets (light curve and period analysis). The project also involves two public workshops, one on observing (already held in August of 2009) and one on data analysis and scientific paper writing (to be held in 2010.) This project has been made possible by the National Science Foundation.

  14. 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...

  15. Creating a high quality factor negative epsilon photonic material from two level resonant systems

    NASA Astrophysics Data System (ADS)

    Thylen, Lars

    2015-02-01

    A densely arranged matrix of passive resonant two-level system is analyzed, aiming at a medium with negative macroscopic epsilon with significantly lower losses than current materials. A quality factor is used to compare the proposed material with prevalent current negative epsilon ones. Requirements are derived on two-level system parameters in terms of spatial dipole density, dipole moment and transversal relaxation times in order to obtain negative epsilon in the range of -2 to -0 and quality factors more than an order of magnitude larger than currently used materials such as silver.

  16. Identification of 14-3-3epsilon substrates from embryonic murine brain.

    PubMed

    Ballif, Bryan A; Cao, Zhongwei; Schwartz, Daniel; Carraway, Kermit L; Gygi, Steven P

    2006-09-01

    Mice deficient in 14-3-3epsilon exhibit abnormal neuronal migration and die perinatally. We report here the first large-scale analysis of 14-3-3 interacting partners from primary animal tissue, identifying from embryonic murine brain 163 14-3-3epsilon interacting proteins and 85 phosphorylation sites on these proteins. Phosphorylation of the deubiquitinating enzyme USP8 at serine 680 was found essential for its interaction with 14-3-3epsilon and for maintaining USP8 in the cytosol. PMID:16944949

  17. Creating a high quality factor negative epsilon photonic material from two level resonant systems

    NASA Astrophysics Data System (ADS)

    Thylen, Lars

    2015-04-01

    A densely arranged matrix of passive resonant two-level system is analyzed, aiming at a medium with negative macroscopic epsilon with significantly lower losses than current materials. A quality factor is used to compare the proposed material with prevalent current negative epsilon ones. Requirements are derived on two-level system parameters in terms of spatial dipole density, dipole moment and transversal relaxation times in order to obtain negative epsilon in the range of -2 to -0 and quality factors more than an order of magnitude larger than currently used materials such as silver.

  18. Clostridium viride sp. nov., a strictly anaerobic bacterium using 5-aminovalerate as growth substrate, previously assigned to Clostridium aminovalericum

    Microsoft Academic Search

    Wolfgang Buckel; Peter H. Janssen; Alexandra Schuhmann; Ulrich Eikmanns; Paul Messner; Uwe Sleytr; Werner Liesack

    1994-01-01

    Strain T2–7, a 5-aminovalerate-fermenting bacterium previously classified as Clostridium aminovalericum, was further characterized, both physiologically and phylogenetically. Comparative sequencing analysis of the almost complete\\u000a 16S rDNA revealed that strain T2–7 forms a distinct lineage within a phylogenetically coherent cluster of gram-positive bacteria\\u000a currently assigned to the genus Clostridium. Strain T2–7 grew with 5-aminovalerate, 5-hydroxyvalerate, 4-hydroxybutyrate, vinylacetate, and crotonate, and required

  19. Characterization of Clostridium thermocellum JW20

    PubMed Central

    Freier, Doris; Mothershed, Cheryle P.; Wiegel, Juergen

    1988-01-01

    Clostridium thermocellum JW20 (ATCC 31549), which was isolated from a Louisiana cotton bale, grew on cellulose, cellobiose, and xylooligomers and, after adaptation, on glucose, fructose, and xylose in the pH range of 7.5 to 6.1 with Topt of 60°C, Tmax of 69°C, and Tmin of above 28°C. Doubling times during growth on cellulose and cellobiose were 6.5 and 2.5 h, respectively. The G+C content of the DNA was 40 mol% (chemical analysis). Growth on cellulose as substrate was totally inhibited in the presence of more than 125 mM sodium sulfate, 300 mM sodium chloride, 250 mM potassium chloride, 200 mM calcium chloride, 125 mM magnesium chloride, 40 mM lactate, or 250 mM acetate. The ratio of the fermentation products ethanol to acetate plus H2 decreased when the culture was agitated. Agitation otherwise increased the rate of cellulose degradation in a growing culture but not under nongrowth conditions or with cell-free culture supernatant containing the extracellular cellulase. Shaking lowered the concentration of H2 in the culture broth and thus minimized inhibition by the H2 formed. Externally added H2 caused an increased formation of ethanol during growth on cellulose or cellobiose. However, at an atmospheric pressure as high as 355 kPa (50 lb/in2), H2 did not cause significant growth inhibition beyond an increasing lag phase (up to 24 h). Several criteria to specifically prove the purity of C. thermocellum cultures were suggested. PMID:16347527

  20. SMIA Geneva September 2005 "Assessing Regional Sustainability with the EPSILON Project"

    E-print Network

    Paris-Sud XI, Université de

    dimension as defined by the UN Commission on Sustainable Development. A coherent objective based structure scale. An attempt to assess sustainability The main foreseen application of the EPSILON model [1 of sustainable development (social, economic, institutional and environmenta