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Sample records for preparations clostridium thermocellum

  1. Promoters and proteins from Clostridium thermocellum and uses thereof

    Science.gov (United States)

    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.

  2. Evaluation of ethanol productivity from cellulose by Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Kurose, N; Yagyu, J; Miyazaki, T; Uchida, M; Hanai, S; Obayashi, A

    1986-01-01

    Clostridium thermocellum, a thermophilic anaerobe, directly converts cellulose to EtOH. To estimate its EtOH production from cellulose, we used a new method based on material balance by which the efficiencies of the enzymes that convert cellulose to ethanol were calculated. Using this method, the maximum efficiency of ethanol production of two strains of C. thermocellum was estimated to be 0.05, with 0.67 as the theoretical maximum. 3 references.

  3. Composition of cellulase complex of Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Golovchenko, N P; Chuvil' skaya, N A; Akimenko, V K

    1985-01-01

    It is thought that the anaerobic thermophilic cellulolytic bacterium C. thermocellum has the potential for direct industrial bioconversion of cellulose into ethanol. Therefore, much attention has been given to the study of the cellulolytic properties of the culture and to the characteristics of the cellulose complex, which is still not completely understood. Hence, the activity and location of various cellulolytic enzymes of C. thermocellum were determined. C. thermocellum has 6 known cellulolytic enzymes. Endoglucanase, cellobiohydrolase and exoglucosidase are extracellular enzymes (99-100 percent of the activity is located outside the cells) while cellulobiases, cellobiose phosphorylase and cellodextrine phosphorylase are inside the cells (80-90% of the activity). 25 references.

  4. Advances in Consolidated Bioprocessing Using Clostridium thermocellum and Thermoanaerobacter saccharolyticum

    Energy Technology Data Exchange (ETDEWEB)

    Lynd, Lee R. [Dartmouth College, Thayer School of Engineering; Guss, Adam M. [ORNL; Himmel, Mike [National Renewable Energy Laboratory (NREL); Beri, Dhananjay [Dartmouth College, Thayer School of Engineering; Herring, Christopher [Mascoma Corporation; Holwerda, Evert [Dartmouth College, Thayer School of Engineering; Murphy, Sean J. [Dartmouth College, Thayer School of Engineering; Olson, Daniel G. [Dartmouth College, Thayer School of Engineering; Paye, Julie [Dartmouth College, Thayer School of Engineering; Rydzak, Thomas [ORNL; Shao, Xiongjun [Dartmouth College, Thayer School of Engineering; Tian, Liang [Dartmouth College, Thayer School of Engineering; Worthen, Robert [Dartmouth College, Thayer School of Engineering

    2016-11-01

    Recent advances are addressed pertaining to consolidated bioprocessing (CBP) of plant cell walls to ethanol using two thermophilic, saccharolytic bacteria: the cellulose-fermenting Clostridium thermocellum and the hemicellulose- fermenting ermoanaerobacterium saccharolyticum. On the basis of the largest comparative dataset assembled to date, it appears that C. thermocellum is substantially more effective at solubilizing unpretreated plant cell walls than industry-standard fungal cellulase, and that this is particularly the case for more recalcitrant feedstocks. e distinctive central metabolism of C. thermocellum appears to involve more extensive energy coupling (e.g., on the order of 5 ATP per glucosyl moiety) than most fermentative anaerobes. Ethanol yields and titers realized by engineered strains of T. saccharolyticum meet standards for industrial feasibility and provide an important proof of concept as well as a model that may be emulated in other organisms. Progress has also been made with C. thermocellum, although not yet to this extent. e current state of strain development is summarized and outstanding challenges for commercial application are discussed. We speculate that CBP organism development is more promising starting with naturally occurring cellulolytic microbes as compared to starting with noncellulolytic hosts.

  5. Metabolic adaption of ethanol-tolerant Clostridium thermocellum.

    Directory of Open Access Journals (Sweden)

    Xinshu Zhu

    Full Text Available Clostridium thermocellum is a major candidate for bioethanol production via consolidated bioprocessing. However, the low ethanol tolerance of the organism dramatically impedes its usage in industry. To explore the mechanism of ethanol tolerance in this microorganism, systematic metabolomics was adopted to analyse the metabolic phenotypes of a C. thermocellum wild-type (WT strain and an ethanol-tolerant strain cultivated without (ET0 or with (ET3 3% (v/v exogenous ethanol. Metabolomics analysis elucidated that the levels of numerous metabolites in different pathways were changed for the metabolic adaption of ethanol-tolerant C. thermocellum. The most interesting phenomenon was that cellodextrin was significantly more accumulated in the ethanol-tolerant strain compared with the WT strain, although cellobiose was completely consumed in both the ethanol-tolerant and wild-type strains. These results suggest that the cellodextrin synthesis was active, which might be a potential mechanism for stress resistance. Moreover, the overflow of many intermediate metabolites, which indicates the metabolic imbalance, in the ET0 cultivation was more significant than in the WT and ET3 cultivations. This indicates that the metabolic balance of the ethanol-tolerant strain was adapted better to the condition of ethanol stress. This study provides additional insight into the mechanism of ethanol tolerance and is valuable for further metabolic engineering aimed at higher bioethanol production.

  6. Potential of thin stillage as a low-cost nutrient source for direct cellulose fermentation by Clostridium thermocellum

    Directory of Open Access Journals (Sweden)

    Rumana Islam

    2015-11-01

    Full Text Available Utilization of thin stillage (TS, derived from grain-based ethanol production, was investigated as an alternative source for microbial growth nutrients during direct conversion of cellulose by Clostridium thermocellum DSM 1237. Fermentation end-products synthesized by C. thermocellum grown on media prepared with various concentrations (50-400 g/L of TS were compared to those synthesized by C. thermocellum grown on reagent grade chemical (reference medium. Cell-growth in TS media, monitored with the aid of quantitative polymerase chain reactions (qPCR technique, showed prolonged growth with increasing TS concentration. Final fermentation end-product concentrations from TS media were comparable with those from the reference medium despite lower growth-rates. The volumetric H2 production generated by C. thermocellum grown with medium containing a low concentration (50 g/L of TS matched the volumetric H2 production by C. thermocellum grown in the reference medium, while higher concentrations (200 g/L of TS resulted in greater synthesis of ethanol. Supplementation of TS-media with Mg++ enhanced ethanol production, while hydrogen production remained unchanged. These results suggest that TS, an attractive source of low-cost nutrients, is capable of supporting the growth of C. thermocellum and that high concentrations of TS favor synthesis of ethanol over hydrogen from cellulose.

  7. How does cellulosome composition influence deconstruction of lignocellulosic substrates in Clostridium (Ruminiclostridium) thermocellum DSM 1313?

    Science.gov (United States)

    Yoav, Shahar; Barak, Yoav; Shamshoum, Melina; Borovok, Ilya; Lamed, Raphael; Dassa, Bareket; Hadar, Yitzhak; Morag, Ely; Bayer, Edward A

    2017-01-01

    Bioethanol production processes involve enzymatic hydrolysis of pretreated lignocellulosic biomass into fermentable sugars. Due to the relatively high cost of enzyme production, the development of potent and cost-effective cellulolytic cocktails is critical for increasing the cost-effectiveness of bioethanol production. In this context, the multi-protein cellulolytic complex of Clostridium ( Ruminiclostridium ) thermocellum, the cellulosome, was studied here. C. thermocellum is known to assemble cellulosomes of various subunit (enzyme) compositions, in response to the available carbon source. In the current study, different carbon sources were used, and their influence on both cellulosomal composition and the resultant activity was investigated. Glucose, cellobiose, microcrystalline cellulose, alkaline-pretreated switchgrass, alkaline-pretreated corn stover, and dilute acid-pretreated corn stover were used as sole carbon sources in the growth media of C. thermocellum strain DSM 1313. The purified cellulosomes were compared for their activity on selected cellulosic substrates. Interestingly, cellulosomes derived from cells grown on lignocellulosic biomass showed no advantage in hydrolyzing the original carbon source used for their production. Instead, microcrystalline cellulose- and glucose-derived cellulosomes were equal or superior in their capacity to deconstruct lignocellulosic biomass. Mass spectrometry analysis revealed differential composition of catalytic and structural subunits (scaffoldins) in the different cellulosome samples. The most abundant catalytic subunits in all cellulosome types include Cel48S, Cel9K, Cel9Q, Cel9R, and Cel5G. Microcrystalline cellulose- and glucose-derived cellulosome samples showed higher endoglucanase-to-exoglucanase ratios and higher catalytic subunit-per-scaffoldin ratios compared to lignocellulose-derived cellulosome types. The results reported here highlight the finding that cellulosomes derived from cells grown on glucose

  8. The ethanol pathway from Thermoanaerobacterium saccharolyticum improves ethanol production in Clostridium thermocellum.

    Science.gov (United States)

    Hon, Shuen; Olson, Daniel G; Holwerda, Evert K; Lanahan, Anthony A; Murphy, Sean J L; Maloney, Marybeth I; Zheng, Tianyong; Papanek, Beth; Guss, Adam M; Lynd, Lee R

    2017-07-01

    Clostridium thermocellum ferments cellulose, is a promising candidate for ethanol production from cellulosic biomass, and has been the focus of studies aimed at improving ethanol yield. Thermoanaerobacterium saccharolyticum ferments hemicellulose, but not cellulose, and has been engineered to produce ethanol at high yield and titer. Recent research has led to the identification of four genes in T. saccharolyticum involved in ethanol production: adhE, nfnA, nfnB and adhA. We introduced these genes into C. thermocellum and observed significant improvements to ethanol yield, titer, and productivity. The four genes alone, however, were insufficient to achieve in C. thermocellum the ethanol yields and titers observed in engineered T. saccharolyticum strains, even when combined with gene deletions targeting hydrogen production. This suggests that other parts of T. saccharolyticum metabolism may also be necessary to reproduce the high ethanol yield and titer phenotype in C. thermocellum. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  9. Improved growth rate in Clostridium thermocellum hydrogenase mutant via perturbed sulfur metabolism.

    Science.gov (United States)

    Biswas, Ranjita; Wilson, Charlotte M; Giannone, Richard J; Klingeman, Dawn M; Rydzak, Thomas; Shah, Manesh B; Hettich, Robert L; Brown, Steven D; Guss, Adam M

    2017-01-01

    Metabolic engineering is a commonly used approach to develop organisms for an industrial function, but engineering aimed at improving one phenotype can negatively impact other phenotypes. This lack of robustness can prove problematic. Cellulolytic bacterium Clostridium thermocellum is able to rapidly ferment cellulose to ethanol and other products. Recently, genes involved in H 2 production, including the hydrogenase maturase hydG and NiFe hydrogenase ech , were deleted from the chromosome of C. thermocellum . While ethanol yield increased, the growth rate of Δ hydG decreased substantially compared to wild type. Addition of 5 mM acetate to the growth medium improved the growth rate in C. thermocellum ∆hydG , whereas wild type remained unaffected. Transcriptomic analysis of the wild type showed essentially no response to the addition of acetate. However, in C. thermocellum ΔhydG , 204 and 56 genes were significantly differentially regulated relative to wild type in the absence and presence of acetate, respectively. Genes, Clo1313_0108-0125, which are predicted to encode a sulfate transport system and sulfate assimilatory pathway, were drastically upregulated in C. thermocellum ΔhydG in the presence of added acetate. A similar pattern was seen with proteomics. Further physiological characterization demonstrated an increase in sulfide synthesis and elimination of cysteine consumption in C. thermocellum ΔhydG . Clostridium thermocellum ΔhydGΔech had a higher growth rate than ΔhydG in the absence of added acetate, and a similar but less pronounced transcriptional and physiological effect was seen in this strain upon addition of acetate. Sulfur metabolism is perturbed in C. thermocellum ΔhydG strains, likely to increase flux through sulfate reduction to act either as an electron sink to balance redox reactions or to offset an unknown deficiency in sulfur assimilation.

  10. Efficient chemoenzymatic oligosaccharide synthesis by reverse phosphorolysis using cellobiose phosphorylase and cellodextrin phosphorylase from Clostridium thermocellum

    DEFF Research Database (Denmark)

    Nakai, Hiroyuki; Abou Hachem, Maher; Petersen, Bent O.

    2010-01-01

    Inverting cellobiose phosphorylase (CtCBP) and cellodextrin phosphorylase (CtCDP) from Clostridium thermocellum ATCC27405 of glycoside hydrolase family 94 catalysed reverse phosphorolysis to produce cellobiose and cellodextrins in 57% and 48% yield from α-d-glucose 1-phosphate as donor with glucose...

  11. Formation and characterization of non-growth states in Clostridium thermocellum: spores and L-forms

    Directory of Open Access Journals (Sweden)

    Mearls Elizabeth B

    2012-08-01

    Full Text Available Abstract Background Clostridium thermocellum is an anaerobic thermophilic bacterium that exhibits high levels of cellulose solublization and produces ethanol as an end product of its metabolism. Using cellulosic biomass as a feedstock for fuel production is an attractive prospect, however, growth arrest can negatively impact ethanol production by fermentative microorganisms such as C. thermocellum. Understanding conditions that lead to non-growth states in C. thermocellum can positively influence process design and culturing conditions in order to optimize ethanol production in an industrial setting. Results We report here that Clostridium thermocellum ATCC 27405 enters non-growth states in response to specific growth conditions. Non-growth states include the formation of spores and a L-form-like state in which the cells cease to grow or produce the normal end products of metabolism. Unlike other sporulating organisms, we did not observe sporulation of C. thermocellum in low carbon or nitrogen environments. However, sporulation did occur in response to transfers between soluble and insoluble substrates, resulting in approximately 7% mature spores. Exposure to oxygen caused a similar sporulation response. Starvation conditions during continuous culture did not result in spore formation, but caused the majority of cells to transition to a L-form state. Both spores and L-forms were determined to be viable. Spores exhibited enhanced survival in response to high temperature and prolonged storage compared to L-forms and vegetative cells. However, L-forms exhibited faster recovery compared to both spores and stationary phase cells when cultured in rich media. Conclusions Both spores and L-forms cease to produce ethanol, but provide other advantages for C. thermocellum including enhanced survival for spores and faster recovery for L-forms. Understanding the conditions that give rise to these two different non-growth states, and the implications that

  12. Clostridium thermocellum: adhesion and sporulation while adhered to cellulose and hemicellulose

    Energy Technology Data Exchange (ETDEWEB)

    Wiegel, J.; Dykstra, M.

    1984-01-01

    During growth in the presence of fibers composed of cellulose or hemicellulose, various strains of the thermophilic soil bacterium Clostridium thermocellum and several newly isolated thermophilic anaerobic soil bacteria adhered to the fibers. Attachment occurred via a fibrous ruthenium red-staining material. C. thermocellum sporulated while attached to the fibers when the pH dropped below 6.4. It is postulated that the attachment is involved in cellulose breakdown and that C. thermocellum gaines an advantage by remaining attached to its insoluble substrates when the environment is not suitable for rapid growth. The tendency to adhere to cellulose fibers was used in the purification of thermophilic cellulolytic anaerobes. 27 references, 7 figures.

  13. Cellulolytic (cel) genes of Clostridium thermocellum F7 and the proteins encoded by them

    International Nuclear Information System (INIS)

    Piruzyan, E.S.; Mogutov, M.A.; Velikodvorskaya, G.A.; Pushkarskaya, T.A.

    1988-01-01

    This study is concerned with genes cell, ce12, and ce13 encoding the endoglucanase of the cellulolytic complex of the anaerobic thermophilic Clostridium thermocellum F7 bacteria, these genes having been closed by us earlier. The authors present the characteristics of proteins synthesized by the cel genes in the minicell system of the strain Escherichia coli K-12 X925. The molecular weights of the proteins encoded by genes cell, ce12, and ce13 are 30,000, 45,000, and 50,000 dalton, respectively. The study of the homology of the cloned section of the C. thermocellum DNA containing the endoglucanase genes, using Southern's blot-hybridization method, did not reveal their physical linkage in the genome. The authors detected a plasmid with a size of about 30 kb in the cells of the C. thermocellum F7 strain investigated

  14. Genome-scale metabolic analysis of Clostridium thermocellum for bioethanol production

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    Brooks J Paul

    2010-03-01

    Full Text Available Abstract Background Microorganisms possess diverse metabolic capabilities that can potentially be leveraged for efficient production of biofuels. Clostridium thermocellum (ATCC 27405 is a thermophilic anaerobe that is both cellulolytic and ethanologenic, meaning that it can directly use the plant sugar, cellulose, and biochemically convert it to ethanol. A major challenge in using microorganisms for chemical production is the need to modify the organism to increase production efficiency. The process of properly engineering an organism is typically arduous. Results Here we present a genome-scale model of C. thermocellum metabolism, iSR432, for the purpose of establishing a computational tool to study the metabolic network of C. thermocellum and facilitate efforts to engineer C. thermocellum for biofuel production. The model consists of 577 reactions involving 525 intracellular metabolites, 432 genes, and a proteomic-based representation of a cellulosome. The process of constructing this metabolic model led to suggested annotation refinements for 27 genes and identification of areas of metabolism requiring further study. The accuracy of the iSR432 model was tested using experimental growth and by-product secretion data for growth on cellobiose and fructose. Analysis using this model captures the relationship between the reduction-oxidation state of the cell and ethanol secretion and allowed for prediction of gene deletions and environmental conditions that would increase ethanol production. Conclusions By incorporating genomic sequence data, network topology, and experimental measurements of enzyme activities and metabolite fluxes, we have generated a model that is reasonably accurate at predicting the cellular phenotype of C. thermocellum and establish a strong foundation for rational strain design. In addition, we are able to draw some important conclusions regarding the underlying metabolic mechanisms for observed behaviors of C. thermocellum

  15. Clostridium thermocellum ATCC27405 transcriptomic, metabolomic and proteomic profiles after ethanol stress

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    Yang Shihui

    2012-07-01

    Full Text Available Abstract Background Clostridium thermocellum is a candidate consolidated bioprocessing biocatalyst, which is a microorganism that expresses enzymes for both cellulose hydrolysis and its fermentation to produce fuels such as lignocellulosic ethanol. However, C. thermocellum is relatively sensitive to ethanol compared to ethanologenic microorganisms such as yeast and Zymomonas mobilis that are used in industrial fermentations but do not possess native enzymes for industrial cellulose hydrolysis. Results In this study, C. thermocellum was grown to mid-exponential phase and then treated with ethanol to a final concentration of 3.9 g/L to investigate its physiological and regulatory responses to ethanol stress. Samples were taken pre-shock and 2, 12, 30, 60, 120, and 240 min post-shock, and from untreated control fermentations for systems biology analyses. Cell growth was arrested by ethanol supplementation with intracellular accumulation of carbon sources such as cellobiose, and sugar phosphates, including fructose-6-phosphate and glucose-6-phosphate. The largest response of C. thermocellum to ethanol shock treatment was in genes and proteins related to nitrogen uptake and metabolism, which is likely important for redirecting the cells physiology to overcome inhibition and allow growth to resume. Conclusion This study suggests possible avenues for metabolic engineering and provides comprehensive, integrated systems biology datasets that will be useful for future metabolic modeling and strain development endeavors.

  16. The identification of four histidine kinases that influence sporulation in Clostridium thermocellum.

    Science.gov (United States)

    Mearls, Elizabeth B; Lynd, Lee R

    2014-08-01

    In this study, we sought to identify genes involved in the onset of spore formation in Clostridium thermocellum via targeted gene deletions, gene over-expression, and transcriptional analysis. We determined that three putative histidine kinases, clo1313_0286, clo1313_2735 and clo1313_1942 were positive regulators of sporulation, while a fourth kinase, clo1313_1973, acted as a negative regulator. Unlike Bacillus or other Clostridium species, the deletion of a single positively regulating kinase was sufficient to abolish sporulation in this organism. Sporulation could be restored in these asporogenous strains via overexpression of any one of the positive regulators, indicating a high level of redundancy between these kinases. In addition to having a sporulation defect, deletion of clo1313_2735 produced L-forms. Thus, this kinase may play an additional role in repressing L-form formation. This work suggests that C. thermocellum enters non-growth states based on the sensory input from multiple histidine kinases. The ability to control the development of non-growth states at the genetic level has the potential to inform strategies for improved strain development, as well as provide valuable insight into C. thermocellum biology. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Clostridium thermocellum Transcriptomic Profiles after Exposure to Furfural or Heat Stress

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, Charlotte M [ORNL; Yang, Shihui [ORNL; Rodriguez, Jr., Miguel [ORNL; Ma, Qin [University of Georgia, Athens, GA; Johnson, Courtney M [ORNL; Dice, Lezlee T [ORNL; Xu, Ying [University of Georgia, Athens, GA; Brown, Steven D [ORNL

    2013-01-01

    Background The thermophilic anaerobe Clostridium thermocellum is a candidate consolidated bioprocessing (CBP)biocatalyst for cellulosic ethanol production. It is capable of both cellulose solubilization and its fermentation to produce lignocellulosic ethanol. Intolerance to stresses routinely encountered during industrial fermentations may hinder the commercial development of this organism. A previous C. thermocellum ethanol stress study showed that largest transcriptomic response was in genes and proteins related to nitrogen uptake and metabolism. Results In this study, C. thermocellum was grown to mid-exponential phase and treated with furfural or heat to a final concentration of 3 g.L-1 or 68 C respectively to investigate general and specific physiological and regulatory stress responses. Samples were taken at 10, 30, 60 and 120 min post-shock, and from untreated control fermentations, for transcriptomic analyses and fermentation product determinations and compared to a published dataset from an ethanol stress study. Urea uptake genes were induced following furfural stress, but not to the same extent as ethanol stress and transcription from these genes was largely unaffected by heat stress. The largest transcriptomic response to furfural stress was genes for sulfate transporter subunits and enzymes in the sulfate assimilatory pathway, although these genes were also affected late in the heat and ethanol stress responses. Lactate production was higher in furfural treated culture, although the lactate dehydrogenase gene was not differentially expressed under this condition. Other redox related genes such as a copy of the rex gene, a bifunctional acetaldehyde-CoA/alcohol dehydrogenase and adjacent genes did show lower expression after furfural stress compared to the control, heat and ethanol fermentation profiles. Heat stress induced expression from chaperone related genes and overlap was observed with the responses to the other stresses. This study suggests the

  18. Deletion of Type I glutamine synthetase deregulates nitrogen metabolism and increases ethanol production in Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Rydzak, Thomas [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Garcia, David [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Stevenson, David M. [Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology; Sladek, Margaret [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Klingeman, Dawn M. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Holwerda, Evert K. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division; Dartmouth College, Hanover, NH (United States). Thayer School of Engineering; Amador-Noguez, Daniel [Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology; Brown, Steven D. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Guss, Adam M. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center

    2017-05-01

    Clostridium thermocellum rapidly deconstructs cellulose and ferments resulting hydrolysis products into ethanol and other products, and is thus a promising platform organism for the development of cellulosic biofuel production via consolidated bioprocessing. And while recent metabolic engineering strategies have targeted eliminating canonical fermentation products (acetate, lactate, formate, and H2), C. thermocellum also secretes amino acids, which has limited ethanol yields in engineered strains to approximately 70% of the theoretical maximum. To decrease amino acid secretion, we attempted to reduce ammonium assimilation by deleting the Type I glutamine synthetase (glnA) in C. thermocellum. Deletion of glnA reduced levels of secreted valine and total amino acids by 53% and 44% respectively, and increased ethanol yields by 53%. RNA-seq analysis revealed that genes encoding the RNF-complex were more highly expressed in ΔglnA and may have a role in improving NADH-availability for ethanol production. While a significant up-regulation of genes involved in nitrogen assimilation and urea uptake suggested that deletion of glnA induces a nitrogen starvation response, metabolomic analysis showed an increase in intracellular glutamine and α-ketoglutarate levels indicative of nitrogen-rich conditions. Here, we propose that deletion of glnA causes deregulation of nitrogen metabolism, leading to overexpression of nitrogen metabolism genes and, in turn, elevated glutamine/α-ketoglutarate levels. Here we demonstrate that perturbation of nitrogen assimilation is a promising strategy to redirect flux from the production of nitrogenous compounds toward biofuels in C. thermocellum.

  19. Deletion of Type I glutamine synthetase deregulates nitrogen metabolism and increases ethanol production in Clostridium thermocellum.

    Science.gov (United States)

    Rydzak, Thomas; Garcia, David; Stevenson, David M; Sladek, Margaret; Klingeman, Dawn M; Holwerda, Evert K; Amador-Noguez, Daniel; Brown, Steven D; Guss, Adam M

    2017-05-01

    Clostridium thermocellum rapidly deconstructs cellulose and ferments resulting hydrolysis products into ethanol and other products, and is thus a promising platform organism for the development of cellulosic biofuel production via consolidated bioprocessing. While recent metabolic engineering strategies have targeted eliminating canonical fermentation products (acetate, lactate, formate, and H 2 ), C. thermocellum also secretes amino acids, which has limited ethanol yields in engineered strains to approximately 70% of the theoretical maximum. To investigate approaches to decrease amino acid secretion, we attempted to reduce ammonium assimilation by deleting the Type I glutamine synthetase (glnA) in an essentially wild type strain of C. thermocellum. Deletion of glnA reduced levels of secreted valine and total amino acids by 53% and 44% respectively, and increased ethanol yields by 53%. RNA-seq analysis revealed that genes encoding the RNF-complex were more highly expressed in ΔglnA and may have a role in improving NADH-availability for ethanol production. While a significant up-regulation of genes involved in nitrogen assimilation and urea uptake suggested that deletion of glnA induces a nitrogen starvation response, metabolomic analysis showed an increase in intracellular glutamine levels indicative of nitrogen-rich conditions. We propose that deletion of glnA causes deregulation of nitrogen metabolism, leading to overexpression of nitrogen metabolism genes and, in turn, elevated glutamine levels. Here we demonstrate that perturbation of nitrogen assimilation is a promising strategy to redirect flux from the production of nitrogenous compounds toward biofuels in C. thermocellum. Copyright © 2017. Published by Elsevier Inc.

  20. Ethanol production by Clostridium thermocellum grown on hydrothermally and organosolv-pretreated lignocellulosic materials

    Energy Technology Data Exchange (ETDEWEB)

    Hoermeyer, H F; Bonn, G; Bobleter, O; Tailliez, P; Millet, J; Girard, H; Aubert, J P

    1988-12-01

    Two strains of the thermophilic anaerobe Clostridium thermocellum, the wild type NCIB 10682 and its ethanol-hyperproductive mutant 647, were tested for their ability to grow on natural lignocellulosic materials (poplar wood, wheat straw) which had been pretreated by either hydrothermolysis or an organosolv process. For both materials and both strains, the dependencies of substrate accessibility on the pretreatment temperature were established in terms of cellulose hydrolysis and of product formation. In addition to the non-pH-controlled shake flask assays, in vitro experiments with cell-free culture supernatant and in vivo cellulolyses under pH regulation in a laboratory fermenter indicated that lignocellulosics pretreated at approx. 230/sup 0/C were degraded efficiently by the Clostridium strains investigated.

  1. Structure of a fibronectin type III-like module from Clostridium thermocellum

    International Nuclear Information System (INIS)

    Alahuhta, Markus; Xu, Qi; Brunecky, Roman; Adney, William S.; Ding, Shi-You; Himmel, Michael E.; Lunin, Vladimir V.

    2010-01-01

    The 1.6 Å resolution structure of a fibronectin type III-like module from Clostridium thermocellum with two molecules in the asymmetric unit is reported. The 1.6 Å resolution structure of a fibronectin type III-like module from Clostridium thermocellum with two molecules in the asymmetric unit is reported. The crystals used for data collection belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 35.43, b = 45.73, c = 107.72 Å, and the structure was refined to an R factor of 0.166. Structural comparisons found over 800 similar structures in the Protein Data Bank. The broad range of different proteins or protein domains with high structural similarity makes it especially demanding to classify these proteins. Previous studies of fibronectin type III-like modules have indicated that they might function as ligand-binding modules, as a compact form of peptide linkers or spacers between other domains, as cellulose-disrupting modules or as proteins that help large enzyme complexes remain soluble

  2. Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Oguntimein, Gbekeloluwa B. [Morgan State Univ., Baltimore, MD (United States); Rodriguez, Jr., Miguel [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Lab., Oak Ridge, TN (United States). BioEnergy Science Center; Dumitrache, Alexandru [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Lab., Oak Ridge, TN (United States). BioEnergy Science Center; Shollenberger, Todd [National Renewable Energy Lab. (NREL), Golden, CO (United States); Decker, Stephen R. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Davison, Brian H. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Lab., Oak Ridge, TN (United States). BioEnergy Science Center; Brown, Steven D. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Lab., Oak Ridge, TN (United States). BioEnergy Science Center; LanzaTech, Inc., Skokie, IL (United States)

    2017-11-09

    Here, to develop and prototype a high-throughput microplate assay to assess anaerobic microorganisms and lignocellulosic biomasses in a rapid, cost-effective screen for consolidated bioprocessing potential. Clostridium thermocellum parent Δhpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentations when compared to the Δhpt strain. A prototype microplate assay system is developed that will facilitate high-throughput bioprospecting for new lignocellulosic biomass types, genetic variants and new microbial strains for bioethanol production.

  3. Mutant alcohol dehydrogenase leads to improved ethanol tolerance in Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Steven D [ORNL; Guss, Adam M [ORNL; Karpinets, Tatiana V [ORNL; Parks, Jerry M [ORNL; Smolin, Nikolai [ORNL; Yang, Shihui [ORNL; Land, Miriam L [ORNL; Klingeman, Dawn Marie [ORNL; Bhandiwad, Ashwini [Thayer School of Engineering at Dartmouth; Rodriguez, Jr., Miguel [ORNL; Raman, Babu [Dow Chemical Company, The; Shao, Xiongjun [Thayer School of Engineering at Dartmouth; Mielenz, Jonathan R [ORNL; Smith, Jeremy C [ORNL; Keller, Martin [ORNL; Lynd, Lee R [Thayer School of Engineering at Dartmouth

    2011-01-01

    Clostridium thermocellum is a thermophilic, obligately anaerobic, Gram-positive bacterium that is a candidate microorganism for converting cellulosic biomass into ethanol through consolidated bioprocessing. Ethanol intolerance is an important metric in terms of process economics, and tolerance has often been described as a complex and likely multigenic trait for which complex gene interactions come into play. Here, we resequence the genome of an ethanol-tolerant mutant, show that the tolerant phenotype is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE), hypothesize based on structural analysis that cofactor specificity may be affected, and confirm this hypothesis using enzyme assays. Biochemical assays confirm a complete loss of NADH-dependent activity with concomitant acquisition of NADPH-dependent activity, which likely affects electron flow in the mutant. The simplicity of the genetic basis for the ethanol-tolerant phenotype observed here informs rational engineering of mutant microbial strains for cellulosic ethanol production.

  4. Specialized activities and expression differences for Clostridium thermocellum biofilm and planktonic cells.

    Science.gov (United States)

    Dumitrache, Alexandru; Klingeman, Dawn M; Natzke, Jace; Rodriguez, Miguel; Giannone, Richard J; Hettich, Robert L; Davison, Brian H; Brown, Steven D

    2017-02-27

    Clostridium (Ruminiclostridium) thermocellum is a model organism for its ability to deconstruct plant biomass and convert the cellulose into ethanol. The bacterium forms biofilms adherent to lignocellulosic feedstocks in a continuous cell-monolayer in order to efficiently break down and uptake cellulose hydrolysates. We developed a novel bioreactor design to generate separate sessile and planktonic cell populations for omics studies. Sessile cells had significantly greater expression of genes involved in catabolism of carbohydrates by glycolysis and pyruvate fermentation, ATP generation by proton gradient, the anabolism of proteins and lipids and cellular functions critical for cell division consistent with substrate replete conditions. Planktonic cells had notably higher gene expression for flagellar motility and chemotaxis, cellulosomal cellulases and anchoring scaffoldins, and a range of stress induced homeostasis mechanisms such as oxidative stress protection by antioxidants and flavoprotein co-factors, methionine repair, Fe-S cluster assembly and repair in redox proteins, cell growth control through tRNA thiolation, recovery of damaged DNA by nucleotide excision repair and removal of terminal proteins by proteases. This study demonstrates that microbial attachment to cellulose substrate produces widespread gene expression changes for critical functions of this organism and provides physiological insights for two cells populations relevant for engineering of industrially-ready phenotypes.

  5. The emergence of Clostridium thermocellum as a high utility candidate for consolidated bioprocessing applications

    Directory of Open Access Journals (Sweden)

    Arthur eRagauskas

    2014-08-01

    Full Text Available First isolated in 1926, Clostridium thermocellum has recently received increased attention as a high utility candidate for use in consolidated bioprocessing applications. These applications, which seek to process lignocellulosic biomass directly into useful products such as ethanol, are gaining traction as economically feasible routes towards the production of fuel and other high value chemical compounds as the shortcomings of fossil fuels become evident. This review evaluates C. thermocellum’s role in this transitory process by highlighting recent discoveries relating to its genomic, transcriptomic, proteomic, and metabolomic responses to varying biomass sources, with a special emphasis placed on providing an overview of its unique, multivariate enzyme cellulosome complex and the role that this structure performs during biomass degradation. Both naturally evolved and genetically engineered strains are examined in light of their unique attributes and responses to various biomass treatment conditions, and the genetic tools that have been employed for their creation are presented. Several future routes for potential industrial usage are presented, and it is concluded that, although there have been many advances to significantly improve C. thermocellum’s amenability to industrial use, several hurdles still remain to be overcome as this unique organism enjoys increased attention within the scientific community.

  6. COMPARATIVE MODELLING AND LIGAND BINDING SITE PREDICTION OF A FAMILY 43 GLYCOSIDE HYDROLASE FROM Clostridium thermocellum

    Directory of Open Access Journals (Sweden)

    Shadab Ahmed

    2012-06-01

    Full Text Available The phylogenetic analysis of Clostridium thermocellum family 43 glycoside hydrolase (CtGH43 showed close evolutionary relation with carbohydrate binding family 6 proteins from C. cellulolyticum, C. papyrosolvens, C. cellulyticum, and A. cellulyticum. Comparative modeling of CtGH43 was performed based on crystal structures with PDB IDs 3C7F, 1YIF, 1YRZ, 2EXH and 1WL7. The structure having lowest MODELLER objective function was selected. The three-dimensional structure revealed typical 5-fold beta–propeller architecture. Energy minimization and validation of predicted model with VERIFY 3D indicated acceptability of the proposed atomic structure. The Ramachandran plot analysis by RAMPAGE confirmed that family 43 glycoside hydrolase (CtGH43 contains little or negligible segments of helices. It also showed that out of 301 residues, 267 (89.3% were in most favoured region, 23 (7.7% were in allowed region and 9 (3.0% were in outlier region. IUPred analysis of CtGH43 showed no disordered region. Active site analysis showed presence of two Asp and one Glu, assumed to form a catalytic triad. This study gives us information about three-dimensional structure and reaffirms the fact that it has the similar core 5-fold beta–propeller architecture and so probably has the same inverting mechanism of action with the formation of above mentioned catalytic triad for catalysis of polysaccharides.

  7. Bioconversion of Agricultural Waste to Ethanol by SSF Using Recombinant Cellulase from Clostridium thermocellum

    Directory of Open Access Journals (Sweden)

    Ruchi Mutreja

    2011-01-01

    Full Text Available The effect of different pretreatment methods, temperature, and enzyme concentration on ethanol production from 8 lignocellulosic agrowaste by simultaneous saccharification and fermentation (SSF using recombinant cellulase and Saccharomyces cerevisiae were studied. Recombinant cellulase was isolated from E. coli BL21 cells transformed with CtLic26A-Cel5-CBM11 full-length gene from Clostridium thermocellum and produced in both batch and fed-batch processes. The maximum cell OD and specific activity in batch mode were 1.6 and 1.91 U/mg, respectively, whereas in the fed-batch mode, maximum cell OD and specific activity were 3.8 and 3.5 U/mg, respectively, displaying a 2-fold increase. Eight substrates, Syzygium cumini (jamun, Azadirachta indica (neem, Saracens indica (asoka, bambusa dendrocalmus (bamboo, Populas nigra (poplar, Achnatherum hymenoides (wild grass, Eucalyptus marginata (eucalyptus, and Mangifera indica (mango, were subjected to SSF. Of three pretreatments, acid, alkali, and steam explosion, acid pretreatment Syzygium cumini (Jamun at 30°C gave maximum ethanol yield of 1.42 g/L.

  8. Bioconversion of Agricultural Waste to Ethanol by SSF Using Recombinant Cellulase from Clostridium thermocellum.

    Science.gov (United States)

    Mutreja, Ruchi; Das, Debasish; Goyal, Dinesh; Goyal, Arun

    2011-01-01

    The effect of different pretreatment methods, temperature, and enzyme concentration on ethanol production from 8 lignocellulosic agrowaste by simultaneous saccharification and fermentation (SSF) using recombinant cellulase and Saccharomyces cerevisiae were studied. Recombinant cellulase was isolated from E. coli BL21 cells transformed with CtLic26A-Cel5-CBM11 full-length gene from Clostridium thermocellum and produced in both batch and fed-batch processes. The maximum cell OD and specific activity in batch mode were 1.6 and 1.91 U/mg, respectively, whereas in the fed-batch mode, maximum cell OD and specific activity were 3.8 and 3.5 U/mg, respectively, displaying a 2-fold increase. Eight substrates, Syzygium cumini (jamun), Azadirachta indica (neem), Saracens indica (asoka), bambusa dendrocalmus (bamboo), Populas nigra (poplar), Achnatherum hymenoides (wild grass), Eucalyptus marginata (eucalyptus), and Mangifera indica (mango), were subjected to SSF. Of three pretreatments, acid, alkali, and steam explosion, acid pretreatment Syzygium cumini (Jamun) at 30°C gave maximum ethanol yield of 1.42 g/L.

  9. Comparison of transcriptional profiles of Clostridium thermocellum grown on cellobiose and pretreated yellow poplar using RNA-Seq

    Directory of Open Access Journals (Sweden)

    Hui eWei

    2014-04-01

    Full Text Available The anaerobic, thermophilic bacterium, Clostridium thermocellum, secretes multi-protein enzyme complexes, termed cellulosomes, which synergistically interact with the microbial cell surface and efficiently disassemble plant cell wall biomass. C. thermocellum has also been considered a potential consolidated bioprocessing (CBP organism due to its ability to produce the biofuel products, hydrogen and ethanol. We found that C. thermocellum fermentation of pretreated yellow poplar (PYP produced 30% and 39% of ethanol and hydrogen product concentrations, respectively, compared to fermentation of cellobiose. RNA-seq was used to analyze the transcriptional profiles of these cells. The PYP-grown cells taken for analysis at the late stationary phase showed 1211 genes up-regulated and 314 down-regulated by more than 2-fold compared to the cellobiose-grown cells. These affected genes cover a broad spectrum of specific functional categories. The transcriptional analysis was further validated by sub-proteomics data taken from the literature; as well as by quantitative reverse transcription-PCR (qRT-PCR analyses of selected genes. Specifically, 47 cellulosomal protein-encoding genes, genes for 4 pairs of SigI-RsgI for polysaccharide sensing, 7 cellodextrin ABC transporter genes, and a set of NAD(PH hydogenase and alcohol dehydrogenase genes were up-regulated for cells growing on PYP compared to cellobiose. These genes could be potential candidates for future studies aimed at gaining insight into the regulatory mechanism of this organism as well as for improvement of C. thermocellum in its role as a CBP organism.

  10. Comparative genotyping of Clostridium thermocellum strains isolated from biogas plants: genetic markers and characterization of cellulolytic potential.

    Science.gov (United States)

    Koeck, Daniela E; Zverlov, Vladimir V; Liebl, Wolfgang; Schwarz, Wolfgang H

    2014-07-01

    Clostridium thermocellum is among the most prevalent of known anaerobic cellulolytic bacteria. In this study, genetic and phenotypic variations among C. thermocellum strains isolated from different biogas plants were determined and different genotyping methods were evaluated on these isolates. At least two C. thermocellum strains were isolated independently from each of nine different biogas plants via enrichment on cellulose. Various DNA-based genotyping methods such as ribotyping, RAPD (Random Amplified Polymorphic DNA) and VNTR (Variable Number of Tandem Repeats) were applied to these isolates. One novel approach - the amplification of unknown target sequences between copies of a previously discovered Random Inserted Mobile Element (RIME) - was also tested. The genotyping method with the highest discriminatory power was found to be the amplification of the sequences between the insertion elements, where isolates from each biogas plant yielded a different band pattern. Cellulolytic potentials, optimal growth conditions and substrate spectra of all isolates were characterized to help identify phenotypic variations. Irrespective of the genotyping method used, the isolates from each individual biogas plant always exhibited identical patterns. This is suggestive of a single C. thermocellum strain exhibiting dominance in each biogas plant. The genotypic groups reflect the results of the physiological characterization of the isolates like substrate diversity and cellulase activity. Conversely, strains isolated across a range of biogas plants differed in their genotyping results and physiological properties. Both strains isolated from one biogas plant had the best specific cellulose-degrading properties and might therefore achieve superior substrate utilization yields in biogas fermenters. Copyright © 2014 Elsevier GmbH. All rights reserved.

  11. Molecular Cloning, Expression and Characterization of Pectin Methylesterase (CtPME) from Clostridium thermocellum.

    Science.gov (United States)

    Rajulapati, Vikky; Goyal, Arun

    2017-05-01

    Many phytopathogenic micro-organisms such as bacteria and fungi produce pectin methylesterases (PME) during plant invasion. Plants and insects also produce PME to degrade plant cell wall. In the present study, a thermostable pectin methylesterase (CtPME) from Clostridium thermocellum belonging to family 8 carbohydrate esterase (CE8) was cloned, expressed and purified. The amino acid sequence of CtPME exhibited similarity with pectin methylesterase from Erwinia chrysanthemi with 38% identity. The gene encoding CtPME was cloned into pET28a(+) vector and expressed using Escherichia coli BL21(DE3) cells. The recombinant CtPME expressed as a soluble protein and exhibited a single band of molecular mass approximately 35.2 kDa on SDS-PAGE gels. The molecular mass, 35.5 kDa of the enzyme, was also confirmed by MALDI-TOF MS analysis. Notably, highest protein concentration (11.4 mg/mL) of CtPME was achieved in auto-induction medium, as compared with LB medium (1.5 mg/mL). CtPME showed maximum activity (18.1 U/mg) against citrus pectin with >85% methyl esterification. The optimum pH and temperature for activity of CtPME were 8.5 and 50 °C, respectively. The enzyme was stable in pH range 8.0-9.0 and thermostable between 45 and 70 °C. CtPME activity was increased by 40% by 5 mM Ca 2+ or Mg 2+ ions. Protein melting curve of CtPME gave a peak at 80 °C. The peak was shifted to 85 °C in the presence of 5 mM Ca 2+ ions, and the addition of 5 mM EDTA shifted back the melting peak to 80 °C. CtPME can be potentially used in food and textile industry applications.

  12. Kinetic modeling of batch fermentation for Populus hydrolysate tolerant mutant and wild type strains of Clostridium thermocellum.

    Science.gov (United States)

    Linville, Jessica L; Rodriguez, Miguel; Mielenz, Jonathan R; Cox, Chris D

    2013-11-01

    The extent of inhibition of two strains of Clostridium thermocellum by a Populus hydrolysate was investigated. A Monod-based model of wild type (WT) and Populus hydrolysate tolerant mutant (PM) strains of the cellulolytic bacterium C. thermocellum was developed to quantify growth kinetics in standard media and the extent of inhibition to a Populus hydrolysate. The PM was characterized by a higher growth rate (μmax=1.223 vs. 0.571 h(-1)) and less inhibition (KI,gen=0.991 vs. 0.757) in 10% v/v Populus hydrolysate compared to the WT. In 17.5% v/v Populus hydrolysate inhibition of PM increased slightly (KI,gen=0.888), whereas the WT was strongly inhibited and did not grow in a reproducible manner. Of the individual inhibitors tested, 4-hydroxybenzoic acid was the most inhibitory, followed by galacturonic acid. The PM did not have a greater ability to detoxify the hydrolysate than the WT. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Pentose sugars inhibit metabolism and increase expression of an AgrD-type cyclic pentapeptide in Clostridium thermocellum.

    Science.gov (United States)

    Verbeke, Tobin J; Giannone, Richard J; Klingeman, Dawn M; Engle, Nancy L; Rydzak, Thomas; Guss, Adam M; Tschaplinski, Timothy J; Brown, Steven D; Hettich, Robert L; Elkins, James G

    2017-02-23

    Clostridium thermocellum could potentially be used as a microbial biocatalyst to produce renewable fuels directly from lignocellulosic biomass due to its ability to rapidly solubilize plant cell walls. While the organism readily ferments sugars derived from cellulose, pentose sugars from xylan are not metabolized. Here, we show that non-fermentable pentoses inhibit growth and end-product formation during fermentation of cellulose-derived sugars. Metabolomic experiments confirmed that xylose is transported intracellularly and reduced to the dead-end metabolite xylitol. Comparative RNA-seq analysis of xylose-inhibited cultures revealed several up-regulated genes potentially involved in pentose transport and metabolism, which were targeted for disruption. Deletion of the ATP-dependent transporter, CbpD partially alleviated xylose inhibition. A putative xylitol dehydrogenase, encoded by Clo1313_0076, was also deleted resulting in decreased total xylitol production and yield by 41% and 46%, respectively. Finally, xylose-induced inhibition corresponds with the up-regulation and biogenesis of a cyclical AgrD-type, pentapeptide. Medium supplementation with the mature cyclical pentapeptide also inhibits bacterial growth. Together, these findings provide new foundational insights needed for engineering improved pentose utilizing strains of C. thermocellum and reveal the first functional Agr-type cyclic peptide to be produced by a thermophilic member of the Firmicutes.

  14. Studies on the enzymology of cellulose degradation by the anaerobic bacterium Clostridium thermocellum and the anaerobic fungus Neocallimastix frontalis

    Energy Technology Data Exchange (ETDEWEB)

    Bhat, K.M.; Gow, L.A.; Wilson, C.A.; Wood, T.W. (Rowett Research Inst., Aberdeen (UK))

    1990-01-01

    The extracellular cellulases from the anaerobic bacterium Clostridium thermocellum and the anaerobic rumen fungus Neocallimastix frontalis are very active on crystalline cellulose. In both cases the activity resides in a high molecular weight complex. The complex from C. thermocellum (termed the cellulosome) was found to be readily dissociated at pH 5.0 and at room temperature by a mixture of SDS, EDTA and DTT. Virtually all the activity of the unfractionated cellulosome was recovered when the dissociated enzyme components were reassociated by dialysis. Thus, the route is now established for the first time for a meaningful study of the mechanism of cellulase action of this commercially important enzyme system. Nearly all of the activity to crystalline cellulose shown by the cellulase of N. frontalis was associated with a fraction which comprised only 2% of the extracellular protein, 3% of the endoglucanase and 3% of the {beta}-glucosidase. This fraction, which could be isolated by affinity chromatography on cellulose, was produced in greater quantity when the fungus was grown in co-culture with the methanogen, Methanobrevibacter smithii. The specific activity of the partially purified enzyme for degradation of crystalline cellulose was several-fold greater than that produced by the aerobic fungus T. reesei, which is being developed world-wide for its commercial potential for converting cellulose to fermentable soluble sugars. The cellulase of N. frontalis clearly has great commercial potential. 39 refs., 19 figs., 22 tabs.

  15. Simultaneous determination of amino acids and carbohydrates in culture media of Clostridium thermocellum by valve-switching ion chromatography.

    Science.gov (United States)

    Fa, Yun; Yang, Haiyan; Ji, Chengshuai; Cui, He; Zhu, Xinshu; Du, Juan; Gao, Jun

    2013-10-10

    An improved method for the simultaneous determination of 20 amino acids and 7 carbohydrates using one-valve switching after injection, ion chromatography, and integrated pulsed amperometric detection is proposed. The resolution of the amino acids and carbohydrates in the cation trap column was investigated. In addition, parameters including flow liquid type, flow rate, concentration, and valve-switch timing were optimized. The method is time-saving, effective, and accurate for the simultaneous separation of amino acids and carbohydrates, with a mean correlation coefficient of >0.99 and repeatability of 0.5-4.6% for eight replicates. The method was successfully applied in the analysis of amino acids and carbohydrates in aseptic media and in extracellular culture media of three phenotypes of Clostridium thermocellum. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs

    Directory of Open Access Journals (Sweden)

    Shuen Hon

    2016-12-01

    Full Text Available Clostridium thermocellum is a promising candidate for ethanol production from cellulosic biomass, but requires metabolic engineering to improve ethanol yield. A key gene in the ethanol production pathway is the bifunctional aldehyde and alcohol dehydrogenase, adhE. To explore the effects of overexpressing wild-type, mutant, and exogenous adhEs, we developed a new expression plasmid, pDGO144, that exhibited improved transformation efficiency and better gene expression than its predecessor, pDGO-66. This new expression plasmid will allow for many other metabolic engineering and basic research efforts in C. thermocellum. As proof of concept, we used this plasmid to express 12 different adhE genes (both wild type and mutant from several organisms. Ethanol production varied between clones immediately after transformation, but tended to converge to a single value after several rounds of serial transfer. The previously described mutant C. thermocellum D494G adhE gave the best ethanol production, which is consistent with previously published results. Keywords: Clostridium Thermocellum, Plasmid, adhE, Structural stability, Gene expression

  17. Crystallization and preliminary diffraction studies of CBM3b of cellobiohydrolase 9A from Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Jindou, Sadanari [Department of Molecular Microbiology and Biotechnology, Tel Aviv University 69978 (Israel); Petkun, Svetlana [Department of Molecular Microbiology and Biotechnology, Tel Aviv University 69978 (Israel); The Daniella Rich Institute for Structural Biology, Tel Aviv University 69978 (Israel); Shimon, Linda [Department of Chemical Research Support, The Weizmann Institute of Science, Rehovot 76100 (Israel); Bayer, Edward A. [Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100 (Israel); Lamed, Raphael; Frolow, Felix, E-mail: mbfrolow@post.tau.ac.il [Department of Molecular Microbiology and Biotechnology, Tel Aviv University 69978 (Israel); The Daniella Rich Institute for Structural Biology, Tel Aviv University 69978 (Israel)

    2007-12-01

    The cloning, expression, purification and crystallization of the CBM3b module of cellobiohydrolase 9A from C. thermocellum is described. The crystals diffract to 2.68 Å. Family 3 carbohydrate-binding modules (CBM3s) are associated with the scaffoldin subunit of the multi-enzyme cellulosome complex and with the family 9 glycoside hydrolases, which are multimodular enzymes that act on plant cell-wall polysaccharides, notably cellulose. Here, the crystallization of CBM3b from cellobiohydrolase 9A is reported. The crystals are tetragonal and belong to space group P4{sub 1} or P4{sub 3}. X-ray diffraction data for CBM3b have been collected to 2.68 Å resolution on beamline ID14-4 at the ESRF.

  18. Overexpression, purification and crystallization of the two C-terminal domains of the bifunctional cellulase ctCel9D-Cel44A from Clostridium thermocellum

    International Nuclear Information System (INIS)

    Najmudin, Shabir; Guerreiro, Catarina I. P. D.; Ferreira, Luís M. A.; Romão, Maria J. C.; Fontes, Carlos M. G. A.; Prates, José A. M.

    2005-01-01

    The two C-terminal domains of the cellulase ctCel9D-Cel44A from C. thermocellum cellulosome have been crystallized in tetragonal space group P4 3 2 1 2 and X-ray diffraction data have been collected to 2.1 and 2.8 Å from native and seleno-l-methionine-derivative crystals, respectively. Clostridium thermocellum produces a highly organized multi-enzyme complex of cellulases and hemicellulases for the hydrolysis of plant cell-wall polysaccharides, which is termed the cellulosome. The bifunctional multi-modular cellulase ctCel9D-Cel44A is one of the largest components of the C. thermocellum cellulosome. The enzyme contains two internal catalytic domains belonging to glycoside hydrolase families 9 and 44. The C-terminus of this cellulase, comprising a polycystic kidney-disease module (PKD) and a carbohydrate-binding module (CBM44), has been crystallized. The crystals belong to the tetragonal space group P4 3 2 1 2, containing a single molecule in the asymmetric unit. Native and seleno-l-methionine-derivative crystals diffracted to 2.1 and 2.8 Å, respectively

  19. Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs).

    Science.gov (United States)

    Hon, Shuen; Lanahan, Anthony A; Tian, Liang; Giannone, Richard J; Hettich, Robert L; Olson, Daniel G; Lynd, Lee R

    2016-12-01

    Clostridium thermocellum is a promising candidate for ethanol production from cellulosic biomass, but requires metabolic engineering to improve ethanol yield. A key gene in the ethanol production pathway is the bifunctional aldehyde and alcohol dehydrogenase, adhE . To explore the effects of overexpressing wild-type, mutant, and exogenous adhE s, we developed a new expression plasmid, pDGO144, that exhibited improved transformation efficiency and better gene expression than its predecessor, pDGO-66. This new expression plasmid will allow for many other metabolic engineering and basic research efforts in C. thermocellum . As proof of concept, we used this plasmid to express 12 different adhE genes (both wild type and mutant) from several organisms. Ethanol production varied between clones immediately after transformation, but tended to converge to a single value after several rounds of serial transfer. The previously described mutant C. thermocellum D494G adhE gave the best ethanol production, which is consistent with previously published results.

  20. Overexpression, purification and crystallization of the two C-terminal domains of the bifunctional cellulase ctCel9D-Cel44A from Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Najmudin, Shabir [REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal); Guerreiro, Catarina I. P. D.; Ferreira, Luís M. A. [CIISA - Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa (Portugal); Romão, Maria J. C. [REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal); Fontes, Carlos M. G. A.; Prates, José A. M., E-mail: japrates@fmv.utl.pt [CIISA - Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa (Portugal); REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal)

    2005-12-01

    The two C-terminal domains of the cellulase ctCel9D-Cel44A from C. thermocellum cellulosome have been crystallized in tetragonal space group P4{sub 3}2{sub 1}2 and X-ray diffraction data have been collected to 2.1 and 2.8 Å from native and seleno-l-methionine-derivative crystals, respectively. Clostridium thermocellum produces a highly organized multi-enzyme complex of cellulases and hemicellulases for the hydrolysis of plant cell-wall polysaccharides, which is termed the cellulosome. The bifunctional multi-modular cellulase ctCel9D-Cel44A is one of the largest components of the C. thermocellum cellulosome. The enzyme contains two internal catalytic domains belonging to glycoside hydrolase families 9 and 44. The C-terminus of this cellulase, comprising a polycystic kidney-disease module (PKD) and a carbohydrate-binding module (CBM44), has been crystallized. The crystals belong to the tetragonal space group P4{sub 3}2{sub 1}2, containing a single molecule in the asymmetric unit. Native and seleno-l-methionine-derivative crystals diffracted to 2.1 and 2.8 Å, respectively.

  1. Purification and crystallization of a trimodular complex comprising the type II cohesin–dockerin interaction from the cellulosome of Clostridium thermocellum

    International Nuclear Information System (INIS)

    Adams, Jarrett J.; Pal, Gour; Yam, Katherine; Spencer, Holly L.; Jia, Zongchao; Smith, Steven P.

    2004-01-01

    A trimodular complex comprising the type II cohesin–dockerin interaction from the cellulosome of C. thermocellum has been purified and crystallized by the hanging-drop vapour-diffusion method. A native crystal and a selenomethionine derivative have been analyzed using X-ray diffraction. The high-affinity calcium-mediated type II cohesin–dockerin interaction is responsible for the attachment of the multi-enzyme cellulose-degrading complex, termed the cellulosome, to the cell surface of the thermophilic anaerobe Clostridium thermocellum. A trimodular 40 kDa complex comprising the SdbA type II cohesin and the the CipA type II dockerin–X module modular pair from the cellulosome of C. thermocellum has been crystallized. The crystals belong to space group P2 1 2 1 2 1 , with unit-cell parameters a = 45.21, b = 52.34, c = 154.69 Å. The asymmetric unit contains one molecule of the protein complex and native and selenomethionine-derivative crystals diffracted to 2.1 and 2.0 Å, respectively

  2. Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-β-d-xylanase 10B in complex with xylohexaose

    Energy Technology Data Exchange (ETDEWEB)

    Najmudin, Shabir, E-mail: shabir@dq.fct.unl.pt [REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal); Pinheiro, Benedita A. [CIISA - Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa (Portugal); Romão, Maria J. [REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal); Prates, José A. M.; Fontes, Carlos M. G. A., E-mail: shabir@dq.fct.unl.pt [CIISA - Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa (Portugal); REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal)

    2008-08-01

    The N-terminal moiety of C. thermocellum endo-1,4-β-d-xylanase 10B, comprising a carbohydrate-binding module (CBM22-1) and a GH10 E337A mutant domain, has been crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3{sub 2}21, contain a dimer in the asymmetric unit and diffract to beyond 2.0 Å resolution. The cellulosome of Clostridium thermocellum is a highly organized multi-enzyme complex of cellulases and hemicellulases involved in the hydrolysis of plant cell-wall polysaccharides. The bifunctional multi-modular xylanase Xyn10B is one of the hemicellulase components of the C. thermocellum cellulosome. The enzyme contains an internal glycoside hydrolase family 10 catalytic domain (GH10) and a C-terminal family 1 carbohydrate esterase domain (CE1). The N-terminal moiety of Xyn10B (residues 32–551), comprising a carbohydrate-binding module (CBM22-1) and the GH10 E337A mutant, was crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3{sub 2}21 and contain a dimer in the asymmetric unit. The crystals diffracted to beyond 2.0 Å resolution.

  3. Lignocellulose fermentation and residual solids characterization for senescent switchgrass fermentation by Clostridium thermocellum in the presence and absence of continuous in situ ball-milling

    Energy Technology Data Exchange (ETDEWEB)

    Balch, Michael L.; Holwerda, Evert K.; Davis, Mark F.; Sykes, Robert W.; Happs, Renee M.; Kumar, Rajeev; Wyman, Charles E.; Lynd, Lee R.

    2017-04-12

    Milling during lignocellulosic fermentation, henceforth referred to as cotreatment, is investigated as an alternative to thermochemical pretreatment as a means of enhancing biological solubilization of lignocellulose. We investigate the impact of milling on soluble substrate fermentation by Clostridium thermocellum with comparison to yeast, document solubilization for fermentation of senescent switchgrass with and without ball milling, and characterize residual solids. Soluble substrate fermentation by C. thermocellum proceeded readily in the presence of continuous ball milling but was completely arrested for yeast. Total fractional carbohydrate solubilization achieved after fermentation of senescent switchgrass by C. thermocellum for 5 days was 0.45 without cotreatment or pretreatment, 0.81 with hydrothermal pretreatment (200 degrees C, 15 minutes, severity 4.2), and 0.88 with cotreatment. Acetate and ethanol were the main fermentation products, and were produced at similar ratios with and without cotreatment. Analysis of solid residues was undertaken using molecular beam mass spectrometry (PyMBMS) and solid-state nuclear magnetic resonance spectroscopy (NMR) in order to provide insight into changes in plant cell walls during processing via various modes. The structure of lignin present in residual solids remaining after fermentation with cotreatment appeared to change little, with substantially greater changes observed for hydrothermal pretreatment - particularly with respect to formation of C-C bonds. The observation of high solubilization with little apparent modification of the residue is consistent with cotreatment enhancing solubilization primarily by increasing the access of saccharolytic enzymes to the feedstock, and C. thermocellum being able to attack all the major linkages in cellulosic biomass provided that these linkages are accessible.

  4. Role of the CipA Scaffoldin Protein in Cellulose Solubilization, as Determined by Targeted Gene Deletion and Complementation in Clostridium thermocellum

    Science.gov (United States)

    Olson, Daniel G.; Giannone, Richard J.; Hettich, Robert L.

    2013-01-01

    The CipA scaffoldin protein plays a key role in the Clostridium thermocellum cellulosome. Previous studies have revealed that mutants deficient in binding or solubilizing cellulose also exhibit reduced expression of CipA. To confirm that CipA is, in fact, necessary for rapid solubilization of crystalline cellulose, the gene was deleted from the chromosome using targeted gene deletion technologies. The CipA deletion mutant exhibited a 100-fold reduction in cellulose solubilization rate, although it was eventually able to solubilize 80% of the 5 g/liter cellulose initially present. The deletion mutant was complemented by a copy of cipA expressed from a replicating plasmid. In this strain, Avicelase activity was restored, although the rate was 2-fold lower than that in the wild type and the duration of the lag phase was increased. The cipA coding sequence is located at the beginning of a gene cluster containing several other genes thought to be responsible for the structural organization of the cellulosome, including olpB, orf2p, and olpA. Tandem mass spectrometry revealed a 10-fold reduction in the expression of olpB, which may explain the lower growth rate. This deletion experiment adds further evidence that CipA plays a key role in cellulose solubilization by C. thermocellum, and it raises interesting questions about the differential roles of the anchor scaffoldin proteins OlpB, Orf2p, and SdbA. PMID:23204466

  5. Heterologous expression and characterization of a putative glycoside hydrolase family 43 arabinofuranosidase from Clostridium thermocellum B8

    NARCIS (Netherlands)

    Camargo, de Brenda R.; Claassens, Nico J.; Quirino, Betania Ferraz; Noronha, Eliane F.; Kengen, Servé W.M.

    2018-01-01

    An extensive list of putative cellulosomal enzymes from C. thermocellum is now available in the public databanks, however, most of these remain unvalidated with regard to their activity and expression control mechanisms. This is particularly true of those enzymes putatively involved in hemicellulose

  6. Development of a Multi-Point Quantitation Method to Simultaneously Measure Enzymatic and Structural Components of the Clostridium thermocellum Cellulosome Protein Complex

    Energy Technology Data Exchange (ETDEWEB)

    Dykstra, Andrew B [ORNL; St. Brice, Lois [Dartmouth College; Rodriguez, Jr., Miguel [ORNL; Raman, Babu [ORNL; Izquierdo, Javier [ORNL; Cook, Kelsey [ORNL; Lynd, Lee R [ORNL; Hettich, Robert {Bob} L [ORNL

    2014-01-01

    Clostridium thermocellum has emerged as a leading bioenergy-relevant microbe due to its ability to solubilize cellulose into carbohydrates, mediated by multi-component membrane-attached complexes termed cellulosomes. To probe microbial cellulose utilization rates, it is desirable to be able to measure the concentrations of saccharolytic enzymes and estimate the total amount of cellulosome present on a mass basis. Current cellulase determination methodologies involve labor-intensive purification procedures and only allow for indirect determination of abundance. We have developed a method using multiple reaction monitoring (MRM-MS) to simultaneously quantitate both enzymatic and structural components of the cellulosome protein complex in samples ranging in complexity from purified cellulosomes to whole cell lysates, as an alternative to a previously-developed enzyme-linked immunosorbent assay (ELISA) method of cellulosome quantitation. The precision of the cellulosome mass concentration in technical replicates is better than 5% relative standard deviation for all samples, indicating high precision for determination of the mass concentration of cellulosome components.

  7. Stoichiometric Assembly of the Cellulosome Generates Maximum Synergy for the Degradation of Crystalline Cellulose, as Revealed by In Vitro Reconstitution of the Clostridium thermocellum Cellulosome.

    Science.gov (United States)

    Hirano, Katsuaki; Nihei, Satoshi; Hasegawa, Hiroki; Haruki, Mitsuru; Hirano, Nobutaka

    2015-07-01

    The cellulosome is a supramolecular multienzyme complex formed by species-specific interactions between the cohesin modules of scaffoldin proteins and the dockerin modules of a wide variety of polysaccharide-degrading enzymes. Cellulosomal enzymes bound to the scaffoldin protein act synergistically to degrade crystalline cellulose. However, there have been few attempts to reconstitute intact cellulosomes due to the difficulty of heterologously expressing full-length scaffoldin proteins. We describe the synthesis of a full-length scaffoldin protein containing nine cohesin modules, CipA; its deletion derivative containing two cohesin modules, ΔCipA; and three major cellulosomal cellulases, Cel48S, Cel8A, and Cel9K, of the Clostridium thermocellum cellulosome. The proteins were synthesized using a wheat germ cell-free protein synthesis system, and the purified proteins were used to reconstitute cellulosomes. Analysis of the cellulosome assembly using size exclusion chromatography suggested that the dockerin module of the enzymes stoichiometrically bound to the cohesin modules of the scaffoldin protein. The activity profile of the reconstituted cellulosomes indicated that cellulosomes assembled at a CipA/enzyme molar ratio of 1/9 (cohesin/dockerin = 1/1) and showed maximum synergy (4-fold synergy) for the degradation of crystalline substrate and ∼2.4-fold-higher synergy for its degradation than minicellulosomes assembled at a ΔCipA/enzyme molar ratio of 1/2 (cohesin/dockerin = 1/1). These results suggest that the binding of more enzyme molecules on a single scaffoldin protein results in higher synergy for the degradation of crystalline cellulose and that the stoichiometric assembly of the cellulosome, without excess or insufficient enzyme, is crucial for generating maximum synergy for the degradation of crystalline cellulose. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. ANALYSIS OF STRUCTURAL ELEMENT OF FAMILY 6 CARBOHYDRATE BINDING MODULE (CTCBM6B OF ALPHA-L-ARABINOFURANOSIDASE FROM CLOSTRIDIUM THERMOCELLUM

    Directory of Open Access Journals (Sweden)

    Shadab Ahmed

    2013-06-01

    Full Text Available The amino acid sequence of a family 6 carbohydrate binding module (CtCBM6B from Clostridium thermocellum alpha-L-arabinofuranosidase showed close evolutionary relationship with some other member of family 6 carbohydrate binding modules. The CD spectrum analysis confirmed the secondary structure prediction of CtCBM6B as both showed beta-sheets (44-48% and random coils (52-54% and no alpha-helix. The hydrogen bonding plot of CtCBM6B showed many segments of parallel and anti-parallel beta-strands which was similar to the secondary structure prediction by PSIPRED VIEW. The three dimensional structure of CtCBM6B generated by MODELLER revealed a typical beta-sandwich architecture at its core, characteristic of beta-jelly roll CBM superfamily. The Ramachandran plot analysis by PROCHECK showed that out of 134 residues, 92.9% were in most favoured region, 6.2% in additionally allowed region and only 0.9% in generously allowed region which indicated a stable conformation of 3D model of CtCBM6B. The docking analysis of CtCBM6B for finding putative ligand binding sites showed that it has high binding affinity for arabinobiose, beta-L-arabinofuranose and beta-D-xylopyranose indicated by lower ligand binding energy (-14.28 kcal mol–1, -12.5 kcal mol–1 and -11.3 kcal mol–1, respectively. CtCBM6B also showed appreciable binding affinity with alpha-D-xylopyranose (–10.8 kcal mol–1, beta-L-arabinopyranose (–10.2 kcal mol-1, alpha-L-arabinopyranose (–10.0 kcal mol–1 and alpha-L-arabinofuranose (–8.75 kcal mol–1. The results indicated that CtCBM6B has high potential for binding arabinan, xylans and substituted xylans.

  9. Lyophilized Carnobacterium divergens AS7 bacteriocin preparation improves performance of broiler chickens challenged with Clostridium perfringens

    DEFF Research Database (Denmark)

    Jozefiak, D; Sip, A; Rutkowski, A

    2012-01-01

    isolates. In total, 480 one-day-old male Ross 308 chicks were randomly assigned to 4 experimental groups (12 replicate pens of 10 birds per treatment). The diets were either nonsupplemented or supplemented with a lyophilized preparation of divercin AS7. On d 18, 19, and 20, half of the birds were...

  10. Bioethanol production involving recombinant C. thermocellum hydrolytic hemicellulase and fermentative microbes.

    Science.gov (United States)

    Das, Saprativ P; Ravindran, Rajeev; Ahmed, Shadab; Das, Debasish; Goyal, Dinesh; Fontes, Carlos M G A; Goyal, Arun

    2012-07-01

    The enhancement of the biomass productivity of Escherichia coli cells harbouring the truncated 903 bp gene designated as glycoside hydrolase family 43 (GH43) from Clostridium thermocellum showing hemicellulase activity along with its further use in simultaneous saccharification and fermentation (SSF) process is described. (Phosphoric acid) H(3)PO(4)-acetone treatment and ammonia fibre expansion (AFEX) were the pretreatment strategies employed on the leafy biomass of mango, poplar, neem and asoka among various substrates owing to their high hemicellulose content. GH43 showed optimal activity at a temperature of 50 °C, pH 5.4 with stability over a pH range of 5.0-6.2. A 4-fold escalation in growth of the recombinant E. coli cells was observed when grown using repeated batch strategy in LB medium supplemented with glucose as co-substrate. Candida shehatae utilizing pentose sugars was employed for bioethanol production. AFEX pretreatment proved to be better over acid-acetone technique. The maximum ethanol concentration (1.44 g/L) was achieved for AFEX pretreated mango (1%, w/v) followed by poplar with an ethanol titre (1.32 g/L) in shake flask experiments. A 1.5-fold increase in ethanol titre (2.11 g/L) was achieved with mango (1%, w/v) in a SSF process using a table top 2-L bioreactor with 1 L working volume.

  11. Clostridium difficile

    NARCIS (Netherlands)

    Bakker, Guido J.; Nieuwdorp, Max

    2017-01-01

    Clostridium difficileinfection (CDI), inflammatory bowel disease (IBD), and metabolic diseases such as obesity, type 2 diabetes (T2D), and nonalcoholic steatohepatitis (NASH). Fecal microbiota transplantation (FMT) is currently tested as a therapeutic option in various diseases and can also help to

  12. Chemoenzymatic Synthesis of beta-D-Glucosides using Cellobiose Phosphorylase from Clostridium thermocellum

    Czech Academy of Sciences Publication Activity Database

    De Winter, K.; Van Renterghem, L.; Wuyts, K.; Pelantová, Helena; Křen, Vladimír; Soetaert, W.; Desmet, T.

    2015-01-01

    Roč. 357, č. 8 (2015), s. 1961-1969 ISSN 1615-4150 R&D Projects: GA MŠk(CZ) LD13042; GA MŠk(CZ) 7E11011 Institutional support: RVO:61388971 Keywords : cellobiose phosphorylase * cross-linked enzyme aggregates * beta-glucosides Subject RIV: CE - Biochemistry Impact factor: 6.453, year: 2015

  13. Biomass Structure and Its Contributions to Recalcitrance During Consolidated Bioprocessing with Clostridium Thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Akinosho, Hannah O. [Georgia Inst. of Technology, Atlanta, GA (United States)

    2017-07-01

    Each study provided compelling reasoning to strongly consider lignin’s part in limiting CBP efficiencies at the laboratory and eventually the industrial scale. To make cellulosic ethanol production feasible with CBP, lignin structure and content must be manipulated with genetic modifications or carefully selected in natural variants to combat these difficulties.

  14. The isolation and characterization of new C. thermocellum strains and the evaluation of multiple anaerobic digestion systems

    Science.gov (United States)

    Lv, Wen

    The overall objective of my research was to improve the efficiencies of bioconversions that produce renewable energy from lignocellulosic biomass. To this end, my studies addressed issues important to two promising strategies: consolidated bioprocessing (CBP) and anaerobic digestion (AD). CBP achieves saccharolytic enzyme production, hydrolysis, and fermentation in a single step and is considered to be the most cost-effective model. Anaerobic bacteria that can be used in CBP are highly desirable. To that end, two thermophilic and cellulolytic bacterial strains were isolated and characterized (Chapter 3). Based on 16S rRNA gene sequence analysis, both strains CS7 and CS8 are closely related to Clostridium thermocellum ATCC 27405. However, they had significantly higher specific cellulase activities and ethanol/acetate ratios than C. thermocellum ATCC 27405. As a result, CS7 and CS8 are two new highly cellulolytic and ethanologenic C. thermocellum strains, with application potentials in research and development of CBP. As some of the most promising AD processes, two temperature-phased AD (TPAD) systems, in comparison with a thermophilic single-stage AD (TSAD) system and a mesophilic two-stage AD (MTAD) system, were studied in treating high-strength dairy cattle manure. The TPAD systems, with the thermophilic digesters acidified (AT-TPAD, Chapter 4) or operated at neutral pH (NT-TPAD, Chapter 5), were optimized at the thermophilic temperature of 50°C and a volume ratio between the thermophilic and the mesophilic digesters of 1:2. Despite similar methane productions, the NT-TPAD system achieved significantly higher volatile solid (VS) removal than the AT-TPAD system and needed no external pH adjustments (Chapter 6). At the same overall OLR, the TSAD system achieved the highest performance, followed by the NT-TPAD and the MTAD systems (Chapter 7). Each digester harbored distinct yet dynamic microbial populations, some of which were significantly correlated or associated

  15. Behavior of Clostridium perfringens at low temperatures

    NARCIS (Netherlands)

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

    2004-01-01

    Refrigerated storage is an important step in the preparation of foods and inadequate storage is one of the main causes of food poisoning outbreaks of Clostridium perfringens. Therefore, growth and germination characteristics of C. perfringens in a temperature range of 3-42 degreesC were determined

  16. Polyclonal Antibody Therapies for Clostridium difficile Infection

    Directory of Open Access Journals (Sweden)

    Michael R. Simon

    2014-10-01

    Full Text Available Clostridium difficile infection has emerged as a growing worldwide health problem. The colitis of Clostridium difficile infection results from the synergistic action of C. difficile secreted toxins A and B upon the colon mucosa. A human monoclonal IgG anti-toxin has demonstrated the ability in combination therapy to reduce mortality in C. difficile challenged hamsters. This antibody is currently in a clinical trial for the treatment of human Clostridium difficile infection. More than one group of investigators has considered using polyclonal bovine colostral antibodies to toxins A and B as an oral passive immunization. A significant proportion of the healthy human population possesses polyclonal antibodies to the Clostridium difficile toxins. We have demonstrated that polyclonal IgA derived from the pooled plasma of healthy donors possesses specificity to toxins A and B and can neutralize these toxins in a cell-based assay. This suggests that secretory IgA prepared from such pooled plasma IgA may be able to be used as an oral treatment for Clostridium difficile infection.

  17. Clostridium Difficile Infections

    Science.gov (United States)

    Clostridium difficile (C. difficile) is a bacterium that causes diarrhea and more serious intestinal conditions such as colitis. Symptoms include Watery ... Loss of appetite Nausea Abdominal pain or tenderness C. difficile is more common in people who need ...

  18. Clostridium XIV Meeting

    Energy Technology Data Exchange (ETDEWEB)

    Lynd, Lee

    2016-08-28

    The 14th biannual Clostridium meeting was held at Dartmouth College from August 28 through 31, 2016. As noted in the meeting program (http://clostridiumxiv.com/wp-content/uploads/2016/09/Clostridium_XIV_program.pdf). the meeting featured 119 registered attendees, 33 oral presentations, 5 of which were given by younger presenters, 40 posters, and 2 keynote presentations, with strong participation by female and international scientists.

  19. Taxonogenomic description of four new Clostridium species isolated from human gut: ‘Clostridium amazonitimonense’, ‘Clostridium merdae’, ‘Clostridium massilidielmoense’ and ‘Clostridium nigeriense’

    Directory of Open Access Journals (Sweden)

    M.T. Alou

    2018-01-01

    Full Text Available Culturomics investigates microbial diversity of the human microbiome by combining diversified culture conditions, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rRNA gene identification. The present study allowed identification of four putative new Clostridium sensu stricto species: ‘Clostridium amazonitimonense’ strain LF2T, ‘Clostridium massilidielmoense’ strain MT26T, ‘Clostridium nigeriense’ strain Marseille-P2414T and ‘Clostridium merdae’ strain Marseille-P2953T, which we describe using the concept of taxonogenomics. We describe the main characteristics of each bacterium and present their complete genome sequence and annotation. Keywords: ‘Clostridium amazonitimonense’, ‘Clostridium massilidielmoense’, ‘Clostridium merdae’, ‘Clostridium nigeriense’, culturomics, emerging bacteria, human microbiota, taxonogenomics

  20. Clostridium difficile Infection

    Science.gov (United States)

    ... TeensRead MoreBMI Calculator Acute BronchitisHigh Blood PressureBursitis of the HipHigh CholesterolExercise-induced UrticariaMicroscopic HematuriaKidney CystsDe Quervain’s Tenosynovitis Home Diseases and Conditions Clostridium difficile (C. diff.) ...

  1. Influence of different yeast cell wall preparations and their components on performance and immune and metabolic pathways in Clostridium perfringens-challenged broiler chicks

    Science.gov (United States)

    A study was conducted to evaluate the influence of purification of yeast cell wall (YCW) preparations on broiler performance, and immunogenic and metabolic pathways under microbial challenge. A total of 240 day-of-hatch chicks were distributed among two battery brooder units (48 pens; 5 birds/pen; ...

  2. Immobilized anaerobic fermentation for bio-fuel production by Clostridium co-culture.

    Science.gov (United States)

    Xu, Lei; Tschirner, Ulrike

    2014-08-01

    Clostridium thermocellum/Clostridium thermolacticum co-culture fermentation has been shown to be a promising way of producing ethanol from several carbohydrates. In this research, immobilization techniques using sodium alginate and alkali pretreatment were successfully applied on this co-culture to improve the bio-ethanol fermentation performance during consolidated bio-processing (CBP). The ethanol yield obtained increased by over 60 % (as a percentage of the theoretical maximum) as compared to free cell fermentation. For cellobiose under optimized conditions, the ethanol yields were approaching about 85 % of the theoretical efficiency. To examine the feasibility of this immobilization co-culture on lignocellulosic biomass conversion, untreated and pretreated aspen biomasses were also used for fermentation experiments. The immobilized co-culture shows clear benefits in bio-ethanol production in the CBP process using pretreated aspen. With a 3-h, 9 % NaOH pretreatment, the aspen powder fermentation yields approached 78 % of the maximum theoretical efficiency, which is almost twice the yield of the untreated aspen fermentation.

  3. Sample preparation in separation of the extracellular chitinolytic enzymes of the human intestinal bacterium Clostridium paraputrificum J4 from the culture fluids

    Czech Academy of Sciences Publication Activity Database

    Tishchenko, Galina; Šimůnek, Jiří; Bartoňová, Hana; Dušková, Jarmila; Dohnálek, Jan; Ponomareva, Evgenia; Tennikova, T.

    2011-01-01

    Roč. 879, č. 22 (2011), s. 2175-2178 ISSN 1570-0232 R&D Projects: GA ČR GA310/09/1407; GA ČR GA525/05/2584 Institutional research plan: CEZ:AV0Z40500505; CEZ:AV0Z50450515 Keywords : sample preparation * culture fluids C. paraputrificum J4 * chitinases Subject RIV: EE - Microbiology, Virology Impact factor: 2.888, year: 2011

  4. Modeling growth of Clostridium perfringens in pea soup during cooling

    NARCIS (Netherlands)

    Jong, de A.E.I.; Beumer, R.R.; Zwietering, M.H.

    2005-01-01

    Clostridium perfringens is a pathogen that mainly causes food poisoning outbreaks when large quantities of food are prepared. Therefore, a model was developed to predict the effect of different cooling procedures on the growth of this pathogen during cooling of food: Dutch pea soup. First, a growth

  5. Diagnosis of Clostridium difficile

    DEFF Research Database (Denmark)

    Jensen, M B F; Olsen, K E P; Nielsen, X C

    2015-01-01

    The diagnosis of Clostridium difficile infection (CDI) requires the detection of toxigenic C. difficile or its toxins and a clinical assessment. We evaluated the performance of four nucleic acid amplification tests (NAATs) detecting toxigenic C. difficile directly from faeces compared to routine...... ribotyping and toxinotyping (TT) were performed on culture-positive samples. In parallel, the samples were analysed by four NAATs; two targeting tcdA or tcdB (illumigene® C. difficile and PCRFast® C. difficile A/B) and two multi-target real-time (RT) PCR assays also targeting cdt and tcdC alleles...... characteristic of epidemic and potentially more virulent PCR ribotypes 027, 066 and 078 (GeneXpert® C. difficile/Epi and an 'in-house RT PCR' two-step algorithm). The multi-target assays were significantly more sensitive compared to routine toxigenic culture (p 

  6. A high-dose preparation of lactobacilli and bifidobacteria in the prevention of antibiotic-associated and Clostridium difficile diarrhoea in older people admitted to hospital: a multicentre, randomised, double-blind, placebo-controlled, parallel arm trial (PLACIDE).

    Science.gov (United States)

    Allen, S J; Wareham, K; Wang, D; Bradley, C; Sewell, B; Hutchings, H; Harris, W; Dhar, A; Brown, H; Foden, A; Gravenor, M B; Mack, D; Phillips, C J

    2013-12-01

    Antibiotic-associated diarrhoea (AAD) occurs most commonly in older people admitted to hospital and within 12 weeks of exposure to broad-spectrum antibiotics. Although usually a mild and self-limiting illness, the 15-39% of cases caused by Clostridium difficile infection [C. difficile diarrhoea (CDD)] may result in severe diarrhoea and death. Previous research has shown that probiotics, live microbial organisms that, when administered in adequate numbers, are beneficial to health, may be effective in preventing AAD and CDD. To determine the clinical effectiveness and cost-effectiveness of a high-dose, multistrain probiotic in the prevention of AAD and CDD in older people admitted to hospital. A multicentre, randomised, double-blind, placebo-controlled, parallel-arm trial. Medical, surgical and elderly care inpatient wards in five NHS hospitals in the UK. Eligible patients were aged ≥ 65 years, were exposed to one or more oral or parenteral antibiotics and were without pre-existing diarrhoeal disorders, recent CDD or at risk of probiotic adverse effects. Out of 17,420 patients screened, 2981 (17.1%) were recruited. Participants were allocated sequentially according to a computer-generated random allocation sequence; 1493 (50.1%) were allocated to the probiotic and 1488 (49.9%) to the placebo arm. Vegetarian capsules containing two strains of lactobacilli and two strains of bifidobacteria (a total of 6 × 10(10) organisms per day) were taken daily for 21 days. The placebo was inert maltodextrin powder in identical capsules. The occurrence of AAD within 8 weeks and CDD within 12 weeks of recruitment was determined by participant follow-up and checking hospital laboratory records by research nurses who were blind to arm allocation. Analysis based on the treatment allocated included 2941 (98.7%) participants. Potential risk factors for AAD at baseline were similar in the two study arms. Frequency of AAD (including CDD) was similar in the probiotic (159/1470, 10

  7. Special Concerns for Seniors: Clostridium difficile

    Science.gov (United States)

    ... and Drugs" Home | Contact Us Special Concerns for Seniors Clostridium difficile - an introduction Clostridium difficile (“C. diff”) ... see APUA’s contribution to CDC’s Vital Signs campaign . Seniors are especially at risk People over the age ...

  8. Clostridium subterminale septicemia in an immunocompetent patient

    OpenAIRE

    Daganou Maria; Kyriakoudi Ann; Moraitou Helen; Pontikis Konstantinos; Avgeropoulou Stavrina; Tripolitsioti Paraskevi; Koutsoukou Antonia

    2016-01-01

    Clostridium subterminale is a Clostridium species that has been rarely isolated in the blood of immunocompromised patients. We report a case of C. subterminale septicemia in an immunocompetent patient who presented with acute mediastinitis following spontaneous esophageal rupture.

  9. Clostridium subterminale septicemia in an immunocompetent patient.

    Science.gov (United States)

    Daganou, Maria; Kyriakoudi, Ann; Moraitou, Helen; Pontikis, Konstantinos; Avgeropoulou, Stavrina; Tripolitsioti, Paraskevi; Koutsoukou, Antonia

    2016-01-01

    Clostridium subterminale is a Clostridium species that has been rarely isolated in the blood of immunocompromised patients. We report a case of C. subterminale septicemia in an immunocompetent patient who presented with acute mediastinitis following spontaneous esophageal rupture.

  10. Proposal to restrict the genus Clostridium Prazmowski to Clostridium butyricum and related species.

    Science.gov (United States)

    Lawson, Paul A; Rainey, Fred A

    2016-02-01

    The genus Clostridium as presently constituted is phylogenetically and phenotypically incoherent. Data from polyphasic taxonomic studies indicate that the genus comprises a collection of very heterogeneous species. Numerous phylogenetic studies, principally based on sequencing of the 16S rRNA gene, indicate that the genus Clostridium should be restricted to Clostridium cluster I as Clostridium sensu stricto . Despite these findings, authors continue to add novel species to the genus Clostridium that do not fall within the radiation of cluster I and the type species Clostridium butyricum , thus perpetuating the confusion associated with the taxonomy of this group. Here, we formally propose that members of the genus Clostridium Prazmowski be restricted to the type species C. butyricum and cluster I species. Eubacterium moniliforme , Eubacterium tarantellae , Sarcina maxima and Sarcina ventriculi should be transferred to the genus Clostridium as Clostridium moniliforme comb. nov., Clostridium tarantellae comb. nov., Clostridium maximum comb. nov. and Clostridium ventriculi comb. nov. A novel genus, Hathewaya gen. nov., is proposed for the species Clostridium histolyticum , Clostridium limosum and Clostridium proteolyticum as Hathewaya histolytica gen. nov. comb. nov., Hathewaya limosa comb. nov. and Hathewaya proteolytica comb. nov. The type species of the genus Hathewaya is Hathewaya histolytica.

  11. Clostridium difficile Infection in Outpatients

    Centers for Disease Control (CDC) Podcasts

    2011-11-07

    Dr. Jon Mark Hirshon, Associate Professor of Emergency Medicine at the University of Maryland School of Medicine, discusses Clostridium difficile infection in outpatients.  Created: 11/7/2011 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 11/21/2011.

  12. Optimizing sporulation of Clostridium perfringens

    NARCIS (Netherlands)

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

    2002-01-01

    Many sporulation media have been developed for Clostridium perfringens, but none stimulates sporulation for all strains. The aim of our experiments was to develop a sporulation method using Duncan and Strong (DS) medium, which supports sporulation of a wide variety of strains. Different inoculation

  13. Clostridium difficile in Retail Meats

    Centers for Disease Control (CDC) Podcasts

    Clostridium difficile is a common cause of diarrhea in healthcare settings but little is known about what causes cases in the community. In this podcast, CDC's Dr. L. Clifford McDonald discusses two papers in the May 2009 edition of Emerging Infectious Diseases that explore whether the organism could be found in meat samples purchased in grocery stores in Arizona and Canada.

  14. Statistical radii associated with amino acids to determine the contact map: fixing the structure of a type I cohesin domain in the Clostridium thermocellum cellulosome

    Science.gov (United States)

    Chwastyk, Mateusz; Poma Bernaola, Adolfo; Cieplak, Marek

    2015-07-01

    We propose to improve and simplify protein refinement procedures through consideration of which pairs of amino acid residues should form native contacts. We first consider 11 330 proteins from the CATH database to determine statistical distributions of contacts associated with a given type of amino acid. The distributions are set across the distances between the α-C atoms that are in contact. Based on this data, we determine typical radii of effective spheres that can be placed on the α-C atoms in order to reconstruct the distribution of the contact lengths. This is done by checking for overlaps with enlarged van der Waals spheres associated with heavy atoms on other amino acids. The resulting contacts can be used to identify non-native contacts that may arise during the time evolution of structure-based models. Here, the radii are used to guide reconstruction of nine missing side chains in a type I cohesin domain with the Protein Data Bank code 1AOH. We first identify the likely missing contacts and then sculpt the corresponding side chains by standard refinement tools to achieve consistency with the expected contact map. One ambiguity in refinement is resolved by determining all-atom conformational energies.

  15. Clostridium subterminale septicemia in an immunocompetent patient

    Directory of Open Access Journals (Sweden)

    Daganou Maria

    2016-01-01

    Full Text Available Clostridium subterminale is a Clostridium species that has been rarely isolated in the blood of immunocompromised patients. We report a case of C. subterminale septicemia in an immunocompetent patient who presented with acute mediastinitis following spontaneous esophageal rupture.

  16. Clostridium difficile in Retail Meats

    Centers for Disease Control (CDC) Podcasts

    2009-04-16

    Clostridium difficile is a common cause of diarrhea in healthcare settings but little is known about what causes cases in the community. In this podcast, CDC's Dr. L. Clifford McDonald discusses two papers in the May 2009 edition of Emerging Infectious Diseases that explore whether the organism could be found in meat samples purchased in grocery stores in Arizona and Canada.  Created: 4/16/2009 by Emerging Infectious Diseases.   Date Released: 4/16/2009.

  17. Nieuwe mogelijkheden bij Clostridium difficile-infecties

    NARCIS (Netherlands)

    van Nood, Els; Keller, Josbert J.; Kuijper, Ed J.; Speelman, Peter

    2013-01-01

    Currently available broad spectrum antibiotics are not sufficiently effective against recurrent Clostridium difficile infections (CDI). Donor faecal microbiota transplantation is a very effective treatment for second and recurrent infection but is time-consuming and requires careful screening of

  18. Comparative pathogenomics of Clostridium tetani.

    Directory of Open Access Journals (Sweden)

    Jonathan E Cohen

    Full Text Available Clostridium tetani and Clostridium botulinum produce two of the most potent neurotoxins known, tetanus neurotoxin and botulinum neurotoxin, respectively. Extensive biochemical and genetic investigation has been devoted to identifying and characterizing various C. botulinum strains. Less effort has been focused on studying C. tetani likely because recently sequenced strains of C. tetani show much less genetic diversity than C. botulinum strains and because widespread vaccination efforts have reduced the public health threat from tetanus. Our aim was to acquire genomic data on the U.S. vaccine strain of C. tetani to better understand its genetic relationship to previously published genomic data from European vaccine strains. We performed high throughput genomic sequence analysis on two wild-type and two vaccine C. tetani strains. Comparative genomic analysis was performed using these and previously published genomic data for seven other C. tetani strains. Our analysis focused on single nucleotide polymorphisms (SNP and four distinct constituents of the mobile genome (mobilome: a hypervariable flagellar glycosylation island region, five conserved bacteriophage insertion regions, variations in three CRISPR (clustered regularly interspaced short palindromic repeats-Cas (CRISPR-associated systems, and a single plasmid. Intact type IA and IB CRISPR/Cas systems were within 10 of 11 strains. A type IIIA CRISPR/Cas system was present in two strains. Phage infection histories derived from CRISPR-Cas sequences indicate C. tetani encounters phages common among commensal gut bacteria and soil-borne organisms consistent with C. tetani distribution in nature. All vaccine strains form a clade distinct from currently sequenced wild type strains when considering variations in these mobile elements. SNP, flagellar glycosylation island, prophage content and CRISPR/Cas phylogenic histories provide tentative evidence suggesting vaccine and wild type strains share a

  19. Comparative pathogenomics of Clostridium tetani.

    Science.gov (United States)

    Cohen, Jonathan E; Wang, Rong; Shen, Rong-Fong; Wu, Wells W; Keller, James E

    2017-01-01

    Clostridium tetani and Clostridium botulinum produce two of the most potent neurotoxins known, tetanus neurotoxin and botulinum neurotoxin, respectively. Extensive biochemical and genetic investigation has been devoted to identifying and characterizing various C. botulinum strains. Less effort has been focused on studying C. tetani likely because recently sequenced strains of C. tetani show much less genetic diversity than C. botulinum strains and because widespread vaccination efforts have reduced the public health threat from tetanus. Our aim was to acquire genomic data on the U.S. vaccine strain of C. tetani to better understand its genetic relationship to previously published genomic data from European vaccine strains. We performed high throughput genomic sequence analysis on two wild-type and two vaccine C. tetani strains. Comparative genomic analysis was performed using these and previously published genomic data for seven other C. tetani strains. Our analysis focused on single nucleotide polymorphisms (SNP) and four distinct constituents of the mobile genome (mobilome): a hypervariable flagellar glycosylation island region, five conserved bacteriophage insertion regions, variations in three CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems, and a single plasmid. Intact type IA and IB CRISPR/Cas systems were within 10 of 11 strains. A type IIIA CRISPR/Cas system was present in two strains. Phage infection histories derived from CRISPR-Cas sequences indicate C. tetani encounters phages common among commensal gut bacteria and soil-borne organisms consistent with C. tetani distribution in nature. All vaccine strains form a clade distinct from currently sequenced wild type strains when considering variations in these mobile elements. SNP, flagellar glycosylation island, prophage content and CRISPR/Cas phylogenic histories provide tentative evidence suggesting vaccine and wild type strains share a common ancestor.

  20. Postpartum Clostridium sordellii infection associated with fatal toxic shock syndrome

    DEFF Research Database (Denmark)

    Rørbye, C; Petersen, Ina Sleimann; Nilas, Lisbeth

    2000-01-01

    Clostridium bacteria are anaerobic Gram positive spore-form-ing bacilli, known to cause distinct clinical syndromes such as botulism, tetanus, pseudomembranous colitis and myonecrosis. The natural habitats of Clostridium species are soil, water and the gastrointestinal tract of animals and humans....... In 5-10% of all women, Clostridium species are also found to be normal inhabitants in the microbial flora of the female genital tract. In case of a non-sexually transmitted genital tract infection, Clostridium species are isolated in 4-20%, and clostridium welchii seems to be the most common isolate....... Clostridium sordellii is rarely encountered in clinical specimens (1% of Clostridium species), but it has been described as a human pathogen with fatal potential. Two toxins, a lethal and a hemorrhagic (that antigenically and pathophysiologically appear similar to Clostridium difficile toxins B and A...

  1. Toxinas de Clostridium perfringens Toxins of Clostridium perfringens

    Directory of Open Access Journals (Sweden)

    W. E. Morris

    2009-12-01

    Full Text Available Clostridium perfringens es un bacilo grampositivo anaerobio con capacidad de formar esporas. Es uno de los patógenos bacterianos con mayor distribución en el medio ambiente, ya que puede ser aislado de muestras de suelo y de agua y además forma parte de la microbiota intestinal de animales y humanos. Sin embargo, en ciertas ocasiones puede actuar como patógeno oportunista y causar enfermedades como la gangrena gaseosa, la enterotoxemia del ovino y del caprino y la disentería del cordero, entre otras. En humanos, está asociado a enfermedades como la intoxicación por alimentos, la enterocolitis necrotizante en niños y la enteritis necrótica o pigbel de las tribus de Papúa-Nueva Guinea. El renovado interés que existe actualmente en el estudio de C. perfringens como patógeno veterinario y humano, junto con el avance de la biología molecular, han hecho posible que la ciencia tenga hoy un conocimiento más profundo sobre la biología y la patogenia de esta bacteria. En esta revisión bibliográfica se discuten y actualizan los principales aspectos de la patogenia intestinal de C. perfringens teniendo en cuenta las toxinas con mayor importancia médica descritas hasta el presente.Clostridium perfringens is an anaerobic gram-positive spore-forming bacillus. It is one of the pathogens with larger distribution in the environment; it can be isolated from soil and water samples, which also belongs to the intestinal flora of animals and humans. However, on some occasions it can act as an opportunistic pathogen, causing diseases such as gas gangrene, enterotoxemia in sheep and goats and lamb dysentery, among others. In human beings, it is associated to diseases such as food poisoning, necrotic enterocolitis of the infant and necrotic enteritis or pigbel in Papua-New Guinea tribes. The renewed interest existing nowadays in the study of C. perfringens as a veterinarian and human pathogen, together with the advance of molecular biology, had enabled

  2. Lactic acid bacteria as protective cultures in fermented pork meat to prevent Clostridium spp. growth.

    Science.gov (United States)

    Di Gioia, Diana; Mazzola, Giuseppe; Nikodinoska, Ivana; Aloisio, Irene; Langerholc, Tomaz; Rossi, Maddalena; Raimondi, Stefano; Melero, Beatriz; Rovira, Jordi

    2016-10-17

    In meat fermented foods, Clostridium spp. growth is kept under control by the addition of nitrite. The growing request of consumers for safer products has led to consider alternative bio-based approaches, the use of protective cultures being one of them. This work is aimed at checking the possibility of using two Lactobacillus spp. strains as protective cultures against Clostridium spp. in pork ground meat for fermented salami preparation. Both Lactobacillus strains displayed anti-clostridia activity in vitro using the spot agar test and after co-culturing them in liquid medium with each Clostridium strain. Only one of them, however, namely L. plantarum PCS20, was capable of effectively surviving in ground meat and of performing anti-microbial activity in carnis in a challenge test where meat was inoculated with the Clostridium strain. Therefore, this work pointed out that protective cultures can be a feasible approach for nitrite reduction in fermented meat products. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Update on Clostridium difficile infections.

    Science.gov (United States)

    Le Monnier, A; Zahar, J-R; Barbut, F

    2014-08-01

    Clostridium difficile infections (CDI) occur primarily in hospitalized patients with risk factors such as concomitant or recent use of antibiotics. CDI related additional costs are important for the global population and health-care facilities. CDI epidemiology has changed since 2003: they became more frequent boosted by large outbreaks, more severe, more resistant to antibiotic treatment, and spread to new groups of population without any risk factor. This is partly due to the emergence and worldwide dissemination of new and more virulent C. difficile strains such as the epidemic clone 027/NAP1/BI. The host immune response plays a central role in the pathogenesis of CDI and could also be involved in the occurrence of recurrent or severe forms. New guidelines including new molecular tests (NAAT) have recently clarified and simplified the diagnostic strategies for the microbiological diagnosis of CDI. The CDI incidence was proven to be related to the level of clinical suspicion and the frequency of microbiological screening for C. difficile. The current recommendations for the treatment of CDI mention oral metronidazole as the first line treatment for mild to moderate diarrhea. Oral vancomycin use should be restricted to severe cases. In the absence of consensus, the treatment of multiple recurrences remains a major concern. New and more targeted antibiotics and innovative therapeutic strategies (fecal transplantation, monoclonal antibodies, and vaccination) have emerged as new therapies for CDI. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  4. Epidemiology of Clostridium difficile Infection

    Science.gov (United States)

    DePestel, Daryl D.; Aronoff, David M.

    2014-01-01

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

  5. EPIDEMIOLOGIC INVESTIGATION OF CLOSTRIDIUM DIFFICILE AND CLOSTRIDIUM PERFRINGENS IN HEALTHY HORSES

    DEFF Research Database (Denmark)

    Schoster, Angelika; Arroyo, Luis; Staempfli, Henry

    Clostridium difficile and Clostridium perfringens are important causes of equine colitis but can also be found in healthy individuals. Epidemiologic information is restricted to cross-sectional studies of fecal shedding with little information on prevalence in gastrointestinal compartments other ...... supports results of previous studies that indicate this organism is rare in healthy horses.......Clostridium difficile and Clostridium perfringens are important causes of equine colitis but can also be found in healthy individuals. Epidemiologic information is restricted to cross-sectional studies of fecal shedding with little information on prevalence in gastrointestinal compartments other...... than feces and variability in shedding over time. The objectives were to investigate the presence of C. difficile and C. perfringens in healthy horses over time and assess prevalence in different gastrointestinal compartments. Feces were collected monthly from 25 horses for one year. Ingesta were...

  6. EGA Protects Mammalian Cells from Clostridium difficile CDT, Clostridium perfringens Iota Toxin and Clostridium botulinum C2 Toxin.

    Science.gov (United States)

    Schnell, Leonie; Mittler, Ann-Katrin; Sadi, Mirko; Popoff, Michel R; Schwan, Carsten; Aktories, Klaus; Mattarei, Andrea; Azarnia Tehran, Domenico; Montecucco, Cesare; Barth, Holger

    2016-04-01

    The pathogenic bacteria Clostridium difficile, Clostridium perfringens and Clostridium botulinum produce the binary actin ADP-ribosylating toxins CDT, iota and C2, respectively. These toxins are composed of a transport component (B) and a separate enzyme component (A). When both components assemble on the surface of mammalian target cells, the B components mediate the entry of the A components via endosomes into the cytosol. Here, the A components ADP-ribosylate G-actin, resulting in depolymerization of F-actin, cell-rounding and eventually death. In the present study, we demonstrate that 4-bromobenzaldehyde N-(2,6-dimethylphenyl)semicarbazone (EGA), a compound that protects cells from multiple toxins and viruses, also protects different mammalian epithelial cells from all three binary actin ADP-ribosylating toxins. In contrast, EGA did not inhibit the intoxication of cells with Clostridium difficile toxins A and B, indicating a possible different entry route for this toxin. EGA does not affect either the binding of the C2 toxin to the cells surface or the enzyme activity of the A components of CDT, iota and C2, suggesting that this compound interferes with cellular uptake of the toxins. Moreover, for C2 toxin, we demonstrated that EGA inhibits the pH-dependent transport of the A component across cell membranes. EGA is not cytotoxic, and therefore, we propose it as a lead compound for the development of novel pharmacological inhibitors against clostridial binary actin ADP-ribosylating toxins.

  7. EGA Protects Mammalian Cells from Clostridium difficile CDT, Clostridium perfringens Iota Toxin and Clostridium botulinum C2 Toxin

    Science.gov (United States)

    Schnell, Leonie; Mittler, Ann-Katrin; Sadi, Mirko; Popoff, Michel R.; Schwan, Carsten; Aktories, Klaus; Mattarei, Andrea; Tehran, Domenico Azarnia; Montecucco, Cesare; Barth, Holger

    2016-01-01

    The pathogenic bacteria Clostridium difficile, Clostridium perfringens and Clostridium botulinum produce the binary actin ADP-ribosylating toxins CDT, iota and C2, respectively. These toxins are composed of a transport component (B) and a separate enzyme component (A). When both components assemble on the surface of mammalian target cells, the B components mediate the entry of the A components via endosomes into the cytosol. Here, the A components ADP-ribosylate G-actin, resulting in depolymerization of F-actin, cell-rounding and eventually death. In the present study, we demonstrate that 4-bromobenzaldehyde N-(2,6-dimethylphenyl)semicarbazone (EGA), a compound that protects cells from multiple toxins and viruses, also protects different mammalian epithelial cells from all three binary actin ADP-ribosylating toxins. In contrast, EGA did not inhibit the intoxication of cells with Clostridium difficile toxins A and B, indicating a possible different entry route for this toxin. EGA does not affect either the binding of the C2 toxin to the cells surface or the enzyme activity of the A components of CDT, iota and C2, suggesting that this compound interferes with cellular uptake of the toxins. Moreover, for C2 toxin, we demonstrated that EGA inhibits the pH-dependent transport of the A component across cell membranes. EGA is not cytotoxic, and therefore, we propose it as a lead compound for the development of novel pharmacological inhibitors against clostridial binary actin ADP-ribosylating toxins. PMID:27043629

  8. Description of Clostridium phoceensis sp. nov., a new species within the genus Clostridium

    Directory of Open Access Journals (Sweden)

    M. Hosny

    2016-11-01

    Full Text Available Clostridium phoceensis sp. nov., strain GD3T (= CSUR P1929 = DSM 100334 is the type strain of C. phoceensis sp. nov., a new species within the genus Clostridium. This strain was isolated from the gut microbiota of a 28-year-old healthy French man. C. phoceensis is a Gram-negative, spore-forming, nonmotile, strictly anaerobic bacterium. We describe its complete genome sequence and annotation, together with its phenotypic characteristics.

  9. Flagellar glycosylation in Clostridium botulinum.

    Science.gov (United States)

    Twine, Susan M; Paul, Catherine J; Vinogradov, Evgeny; McNally, David J; Brisson, Jean-Robert; Mullen, James A; McMullin, David R; Jarrell, Harold C; Austin, John W; Kelly, John F; Logan, Susan M

    2008-09-01

    Flagellins from Clostridium botulinum were shown to be post-translationally modified with novel glycan moieties by top-down MS analysis of purified flagellin protein from strains of various toxin serotypes. Detailed analyses of flagellin from two strains of C. botulinum demonstrated that the protein is modified by a novel glycan moiety of mass 417 Da in O-linkage. Bioinformatic analysis of available C. botulinum genomes identified a flagellar glycosylation island containing homologs of genes recently identified in Campylobacter coli that have been shown to be responsible for the biosynthesis of legionaminic acid derivatives. Structural characterization of the carbohydrate moiety was completed utilizing both MS and NMR spectroscopy, and it was shown to be a novel legionaminic acid derivative, 7-acetamido-5-(N-methyl-glutam-4-yl)-amino-3,5,7,9-tetradeoxy-D-glycero-alpha-D-galacto-nonulosonic acid, (alphaLeg5GluNMe7Ac). Electron transfer dissociation MS with and without collision-activated dissociation was utilized to map seven sites of O-linked glycosylation, eliminating the need for chemical derivatization of tryptic peptides prior to analysis. Marker ions for novel glycans, as well as a unique C-terminal flagellin peptide marker ion, were identified in a top-down analysis of the intact protein. These ions have the potential for use in for rapid detection and discrimination of C. botulinum cells, indicating botulinum neurotoxin contamination. This is the first report of glycosylation of Gram-positive flagellar proteins by the 'sialic acid-like' nonulosonate sugar, legionaminic acid.

  10. Clostridium difficile infection in Thailand.

    Science.gov (United States)

    Putsathit, Papanin; Kiratisin, Pattarachai; Ngamwongsatit, Puriya; Riley, Thomas V

    2015-01-01

    Clostridium difficile is the aetiological agent in ca. 20% of cases of antimicrobial-associated diarrhoea in hospitalised adults. Diseases caused by this organism range from mild diarrhoea to occasional fatal pseudomembranous colitis. The epidemiology of C. difficile infection (CDI) has changed notably in the past decade, following epidemics in the early 2000s of PCR ribotype (RT) 027 infection in North America and Europe, where there was an increase in disease severity and mortality. Another major event has been the emergence of RT 078, initially as the predominant ribotype in production animals in the USA and Europe, and then in humans in Europe. Although there have been numerous investigations of the epidemiology of CDI in North America and Europe, limited studies have been undertaken elsewhere, particularly in Asia. Antimicrobial exposure remains the major risk factor for CDI. Given the high prevalence of indiscriminate and inappropriate use of antimicrobials in Asia, it is conceivable that CDI is relatively common among humans and animals. This review describes the level of knowledge in Thailand regarding C. difficile detection methods, prevalence and antimicrobial susceptibility profile, as well as the clinical features of, treatment options for and outcomes of the disease. In addition, antimicrobial usage in livestock in Thailand will be reviewed. A literature search yielded 18 studies mentioning C. difficile in Thailand, a greater number than from any other Asian country. It is possible that the situation in Thailand in relation to CDI may mirror the situation in other developing Asians countries. Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  11. Clostridium difficile and pediatric inflammatory bowel disease

    DEFF Research Database (Denmark)

    Martinelli, Massimo; Strisciuglio, Caterina; Veres, Gabor

    2014-01-01

    BACKGROUND: Clostridium difficile infection is associated with pediatric inflammatory bowel disease (IBD) in several ways. We sought to investigate C. difficile infection in pediatric patients with IBD in comparison with a group of children with celiac disease and to evaluate IBD disease course o...

  12. Clostridium cadaveris bacteraemia: two cases and review.

    NARCIS (Netherlands)

    Schade, R.P.; Rijn, M. Van; Timmers, H.J.L.M.; Dofferhoff, A.S.M.; Klaassen, C.H.W.; Meis, J.F.G.M.

    2006-01-01

    Clostridium cadaveris is a strict anaerobic Gram-positive rod that is the most prominent bacterium during the decay of dead bodies. We present 2 rare cases of bacteraemia with C. cadaveris. The source of both infectious episodes was most probably of gastrointestinal origin.

  13. Clostridium difficile infection : epidemiology, complications and recurrences

    NARCIS (Netherlands)

    Bauer, Martijn Philippe

    2014-01-01

    Clostridium difficile is a spore-forming bacterium, the toxin-producing strains of which cause colitis. Risk factors are antibiotics, advanced age and severe comorbidity. C. difficile infection (CDI) has been regarded as mostly a hospital-acquired infection. Preventing relapses is considered the

  14. The changing epidemiology of Clostridium difficile infections

    NARCIS (Netherlands)

    Freeman, J.; Bauer, M. P.; Baines, S. D.; Corver, J.; Fawley, W. N.; Goorhuis, B.; Kuijper, E. J.; Wilcox, M. H.

    2010-01-01

    The epidemiology of Clostridium difficile infection (CDI) has changed dramatically during this millennium. Infection rates have increased markedly in most countries with detailed surveillance data. There have been clear changes in the clinical presentation, response to treatment, and outcome of CDI.

  15. Clostridium difficile infection in returning travellers

    NARCIS (Netherlands)

    Michal Stevens, A.; Esposito, Douglas H.; Stoney, Rhett J.; Hamer, Davidson H.; Flores-Figueroa, Jose; Bottieau, Emmanuel; Connor, Bradley A.; Gkrania-Klotsas, Effrossyni; Goorhuis, Abraham; Hynes, Noreen A.; Libman, Michael; Lopez-Velez, Rogelio; McCarthy, Anne E.; von Sonnenburg, Frank; Schwartz, Eli; van Genderen, Perry J. J.; Scott Benson, L.; Leung, Daniel T.

    2017-01-01

    There is increasing recognition of the contribution of community-acquired cases to the global burden of Clostridium difficile infection (CDI). The epidemiology of CDI among international travellers is poorly understood, and factors associated with international travel, such as antibiotic use and

  16. TREATMENT OF CLOSTRIDIUM DIFFICILE- ASSOCIATED DISEASE

    Directory of Open Access Journals (Sweden)

    Snezana Antic-Mladenovic

    2007-04-01

    Full Text Available Clostridium difficile is a Gram-positive, spore-forming, anaerobic bacillus that is widely distributed in the environment, but is found as a part of a normal large bowel flora in approximately 3% of normal adults. C. difficile produces two protein exotoxins: toxin A and toxin B. Both toxins are responsible for causing the sings and symptoms of disease.C. difficile is now thought to be responsible for a spectrum of diseases, ranging from asymptomatic colonization to diarrhea of varying severity, life-threatening colitis, often as a consequence of long-term antibiotic exposure. This spectrum has become known as C. difficile-associated disease (CDAD.Treatment of Clostridium difficile-associated disease demand administration of effi-cient antibiotics (vancomycin, metronidazole, anion exchange resins and probiotics (Lactobacillus spp., Saccharomyces boulardii.

  17. Cellular Entry of Clostridium perfringens Iota-Toxin and Clostridium botulinum C2 Toxin

    Directory of Open Access Journals (Sweden)

    Masaya Takehara

    2017-08-01

    Full Text Available Clostridium perfringens iota-toxin and Clostridium botulinum C2 toxin are composed of two non-linked proteins, one being the enzymatic component and the other being the binding/translocation component. These latter components recognize specific receptors and oligomerize in plasma membrane lipid-rafts, mediating the uptake of the enzymatic component into the cytosol. Enzymatic components induce actin cytoskeleton disorganization through the ADP-ribosylation of actin and are responsible for cell rounding and death. This review focuses upon the recent advances in cellular internalization of clostridial binary toxins.

  18. Cellular Entry of Clostridium perfringens Iota-Toxin and Clostridium botulinum C2 Toxin.

    Science.gov (United States)

    Takehara, Masaya; Takagishi, Teruhisa; Seike, Soshi; Oda, Masataka; Sakaguchi, Yoshihiko; Hisatsune, Junzo; Ochi, Sadayuki; Kobayashi, Keiko; Nagahama, Masahiro

    2017-08-11

    Clostridium perfringens iota-toxin and Clostridium botulinum C2 toxin are composed of two non-linked proteins, one being the enzymatic component and the other being the binding/translocation component. These latter components recognize specific receptors and oligomerize in plasma membrane lipid-rafts, mediating the uptake of the enzymatic component into the cytosol. Enzymatic components induce actin cytoskeleton disorganization through the ADP-ribosylation of actin and are responsible for cell rounding and death. This review focuses upon the recent advances in cellular internalization of clostridial binary toxins.

  19. New techniques for growing anaerobic bacteria: experiments with Clostridium butyricum and Clostridium acetobutylicum

    International Nuclear Information System (INIS)

    Adler, H.I.; Crow, W.D.; Hadden, C.T.; Hall, J.; Machanoff, R.

    1983-01-01

    Stable membrane fragments derived from Escherichia coli produce and maintain strict anaerobic conditions when added to liquid or solid bacteriological media. Techniques for growing Clostridium butyricum and Clostridium acetobutylicum in membrane-containing media are described. Liquid cultures initiated by very small inocula can be grown in direct contact with air. In solid media, colonies develop rapidly from individual cells even without incubation in anaerobic jars or similar devices. Observations on growth rates, spontaneous mutations, radiation, and oxygen sensitivity of anaerobic bacteria have been made using these new techniques

  20. Clostridium botulinum Spores Found in Honey from Small Apiaries in Poland

    Directory of Open Access Journals (Sweden)

    Wojtacka Joanna

    2016-12-01

    Full Text Available A total of 102 honey samples collected from small apiaries (≤ 20 hives in Poland were analysed for the presence of Clostridium botulinum spores. The samples were prepared using the dilution centrifugation method and cultured in parallel in cooked meat medium (CMM and tripticase peptone glucose yeast (TPGY enrichment broths. Identification of toxin types A, B, and E of Clostridium botulinum strains was performed with the use of the multiplex PCR method. Positive samples were also subjected to quantitative analysis with the use of Clostridium botulinum Isolation Agar Base (CBAB. The prevalence analysis showed 22 (21.6% samples contaminated with C. botulinum spores. The major serotype detected was botulin neurotoxin type A – 16 (72.7% whereas type B was found in 3 (13.6% honey samples and type E also only in 3 (13.6% honey samples. Dual-toxin-producing strains were noted. The average quantity of spores in PCR - C. botulinum positive samples was 190 in 1 gram of honey.

  1. Degradation of cellulosic biomass and its subsequent utilization for the production of chemical feedstocks

    Energy Technology Data Exchange (ETDEWEB)

    Wang, D.I.C.; Cooney, C.L.; Demain, A.L.; Gomez, R.F.; Sinskey, A.J.

    1977-11-01

    Progress in studies on the production of reducing sugars and other products by Clostridium thermocellum on cellulosic biomass is reported. The rate of reducing sugar production using corn residue was found to be equal if not greater than on solka floc. Current work is being devoted towards elucidating discrepancies between reducing sugar analysis and high pressure liquid chromatography sugar analysis in order to permit accurate material balances to be completed. Studies are reported in further characterizing the plasmics of C. thermocellum and in the development of protoplasts of the same microorganism. A process and economic analysis for the production of 200 x 10/sup 6/ pounds (90 x 10/sup 6/ kilograms) per year of soluble reducing sugars from corn stover cellulose, using enzymes derived from Clostridium thermocellum was designed. Acrylic acid was produced in resting cell preparation of Clostridium propionicum from both ..beta..-alanine and from propionic acid. Results from the conversion of corn stover hydrolyzates to lactic acid, a precursor to acrylic acid, show that up to 70% of the sugars produced are converted to lactic acid. Efforts are proceeding to improve the conversion yield and carry out the overall conversion of corn stover to acrylic acid in the same fermentor. Results on the production of acetone and butanol by Clostridium acetobutylicum demonstrated the capability of the strain to produce mixed solvents in concentration and conversion similar to that achieved in industrial processes. Various studies on the production of acetic acid by Clostridium thermoaceticum are also reported.

  2. Management of Clostridium difficile diarrhoea in District General ...

    African Journals Online (AJOL)

    ... four cases of Clostridium difficile in our hospital over duration of three months. We looked into the demographic features of the patient population and compliance with the Trust guidelines for the management of the diarrhoea. Keywords:Diarrhoea, Clostridium difficile, Management. Internet Journal of Medical Update Vol.

  3. Clostridium difficile: A healthcare-associated infection of unknown ...

    African Journals Online (AJOL)

    Clostridium difficile: A healthcare-associated infection of unknown significance in adults in sub-Saharan Africa. ... Abstract. Background: Clostridium difficile infection (CDI) causes a high burden of disease in high-resource healthcare systems, with significant morbidity, mortality, and financial implications. CDI is a ...

  4. Plasmidome interchange between Clostridium botulinum, Clostridium novyi and Clostridium haemolyticum converts strains of independent lineages into distinctly different pathogens.

    Science.gov (United States)

    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.

  5. Plasmidome interchange between Clostridium botulinum, Clostridium novyi and Clostridium haemolyticum converts strains of independent lineages into distinctly different pathogens.

    Directory of Open Access Journals (Sweden)

    Hanna Skarin

    Full Text Available 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.

  6. The History of Collagenase Clostridium Histolyticum.

    Science.gov (United States)

    Yang, Kevin K; Bennett, Nelson

    2015-10-01

    After its U.S. FDA approval in 2013, Collagenase Clostridium histolyticum (CCh) has seen increasing use as a nonoperative treatment for Peyronie's disease (PD). We review the history of CCh and trials that led to its adoption. To provide a historical and contemporary context for the evolution of Collagenase Clostridium histolyticum as a treatment modality for Peyronie's disease. A comprehensive search of peer-reviewed literature was performed pertaining to CCh and its biochemical and clinical significance. The main outcome studied was the efficacy and safety profile of CCh in PD. CCh use in other diseases processes and its associated outcomes are also described. CCh injection yields objective improvement in penile curvature across multiple trials in PD patients. Recently, level 1 strength of evidence has emerged supporting its widespread use. As such, CCh stands as the only FDA-approved injectable therapy for PD. Adverse events were namely limited to local reactions. Serious systemic complications and need for intervention were rare. CCh is a safe and effective treatment for PD patients with deformities and plaque configuration amenable to injectable therapy. Multiple trials have demonstrated improvements in objective and subjective metrics such as penile curvature and bother scores. However, multiyear follow-up is needed to assess durability and its sustained clinical significance. Currently, refinement in dosing and technique has established a niche for CCh in PD patients who are affected by their symptoms but are not yet committed to surgical intervention. Yang KK and Bennett N. The history of collagenase clostridium histolyticum. Copyright © 2015 International Society for Sexual Medicine. Published by Elsevier Inc. All rights reserved.

  7. Clostridium Difficile Infection in the Nephrology Ward

    Directory of Open Access Journals (Sweden)

    Sylwia Dudzicz

    2017-11-01

    Full Text Available Clostridium difficile is currently the most frequently identified pathogen causing antibiotic-associated diarrhea and the main cause of nosocomial diarrhea. In recent years, increases incidence of infection, severe infection, recurrent infection and mortality from Clostridium difficile infection (CDI have been observed. This may be a consequence of excessive antibiotic use and spread of the hypervirulent epidemic BI/NAP1/027 strain of Clostridium difficile. The main risk factors for CDI are: antibiotic therapy, previous hospitalizations and number of comorbid conditions. Prevention of CDI mainly is focused in two directions: reducing the exposure of patients to the disease pathogen by intensifying hygiene measures, and reducing the impact of risk factors. A meta-analyses of clinical studies (observational, cohort and case control showed significantly higher risk of CDI and CDI recurrence in patients with chronic kidney disease and increased mortality risk in chronic kidney disease patients with CDI comparing those without CDI. Increased risk of CDI in patients with chronic kidney disease can be caused by: frequent antibiotic therapy associated with numerous infections resulting in intestinal microflora dysfunction, frequent hospitalizations, older age of the patients and an impaired immune system. Among preventative measures against CDI, the use of probiotics were also studied. In patients hospitalized in nephrology ward highly significant reduction of the CDI incidence was observed after the introduction of Lactobacillus plantarum 299v as CDI prophylaxis. Therefore, the use of Lactobacillus plantarum 299v seems to be a promising method of CDI prevention in chronic kidney disease patients hospitalized in nephrology ward.

  8. Conditions associated with Clostridium sporogenes growth as a surrogate for Clostridium botulinum in nonthermally processed canned butter.

    Science.gov (United States)

    Taylor, R H; Dunn, M L; Ogden, L V; Jefferies, L K; Eggett, D L; Steele, F M

    2013-05-01

    The objective of this study was to better understand the effect of butter composition and emulsion structure on growth and survival of Clostridium sporogenes, used as a surrogate for C. botulinum in canned butter. The lack of a thermal process step in commercially available canned butter raises questions of potential safety, because it is hermetically sealed and generally exhibits anaerobic growth conditions, which are optimal for Clostridium botulinum growth. Without thermal processing, low-acid canned foods must have inhibitory factors present to prevent C. botulinum growth. Some potential intrinsic inhibitory factors, or hurdles, within butter include: reduced water activity, acidity in cultured products, elevated salt content, and the micro-droplet nature of the aqueous phase in the butter emulsion. It was hypothesized that a normal, intact butter emulsion would have sufficient hurdles to prevent C. botulinum growth, whereas a broken butter emulsion would result in a coalesced aqueous phase that would allow for C. botulinum growth. Batch-churned butter was inoculated with C. sporogenes; butter samples with varying salt contents (0, 0.8, 1.6, and 2.4% wt/wt NaCl) were prepared and stored in coated steel cans for varying times (1 or 2 wk) and temperatures (22 or 41°C) to determine temperature and emulsion structure effects on C. sporogenes growth. Samples stored at 41°C showed a significant increase in C. sporogenes growth compared with those stored at 22°C. Furthermore, NaCl addition was found to have a significant effect on C. sporogenes growth, with 0.8% NaCl promoting more growth than 0%, but with decreases in growth observed at 1.6 and 2.4%. Uninoculated control plates were also found to have bacterial growth; this growth was attributed to other anaerobic bacteria present within the cream. It was concluded that removal of the hurdle created by the micro-droplet size of the emulsion aqueous phase could result in C. botulinum growth even at elevated salt

  9. Probiotics and prevention of Clostridium difficile infection.

    Science.gov (United States)

    Goldstein, E J C; Johnson, S J; Maziade, P-J; Evans, C T; Sniffen, J C; Millette, M; McFarland, L V

    2017-06-01

    The role of probiotics as adjunctive measures in the prevention of Clostridium difficile infection (CDI) has been controversial. However, a growing body of evidence has suggested that they have a role in primary prevention of CDI. Elements of this controversy are reviewed and the proposed mechanisms of action, the value and cost effectiveness of probiotics are addressed with a focus on three agents, Saccharomyces boulardii, Lactobacillus rhamnosus GG and the combination of Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, Lactobacillus rhamnosus CLR2 (Bio-K+). Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Annotation of the Clostridium Acetobutylicum Genome

    Energy Technology Data Exchange (ETDEWEB)

    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.

  11. Clostridium difficile in Humans and Food Animals

    Centers for Disease Control (CDC) Podcasts

    2008-06-30

    Clostridium difficile is an antibiotic-resistant bacterium that causes diarrhea and sometimes serious intestinal illnesses. In recent years, C. difficile infections have been increasing in number and severity, including among some people outside healthcare settings. In this podcast, CDC's Dr. Michael Jhung discusses his recent study that looked at a new, increasingly prevalent strain of C. difficile in people and compared it to a strain historically found in animals to see whether the two might be linked. The study is published in the July 2008 issue of Emerging Infectious Diseases.  Created: 6/30/2008 by Emerging Infectious Diseases.   Date Released: 7/3/2008.

  12. Clostridium difficile in Humans and Food Animals

    Centers for Disease Control (CDC) Podcasts

    Clostridium difficile is an antibiotic-resistant bacterium that causes diarrhea and sometimes serious intestinal illnesses. In recent years, C. difficile infections have been increasing in number and severity, including among some people outside healthcare settings. In this podcast, CDC's Dr. Michael Jhung discusses his recent study that looked at a new, increasingly prevalent strain of C. difficile in people and compared it to a strain historically found in animals to see whether the two might be linked. The study is published in the July 2008 issue of Emerging Infectious Diseases.

  13. Mortality and Clostridium difficile infection in an Australian setting.

    Science.gov (United States)

    Mitchell, Brett G; Gardner, Anne; Hiller, Janet E

    2013-10-01

    To quantify the risk of death associated with Clostridium difficile infection, in an Australian tertiary hospital. Two reviews examining Clostridium difficile infection and mortality indicate that Clostridium difficile infection is associated with increased mortality in hospitalized patients. Studies investigating the mortality of Clostridium difficile infection in settings outside of Europe and North America are required, so that the epidemiology of Clostridium difficile infection in these regions can be understood and appropriate prevention strategies made. An observational non-concurrent cohort study design was used. Data from all persons who had (exposed) and a matched sample of persons who did not have Clostridium difficile infection, for the calendar years 2007-2010, were analysed. The risk of dying within 30, 60, 90 and 180 days was compared using the two groups. Kaplan-Meier survival analysis and conditional logistic regression models were applied to the data to examine time to death and mortality risk adjusted for comorbidities using the Charlson Comorbidity Index. One hundred and fifty-eight cases of infection were identified. A statistically significant difference in all-cause mortality was identified between exposed and non-exposed groups at 60 and 180 days. In a conditional regression model, mortality in the exposed group was significantly higher at 180 days. In this Australian study, Clostridium difficile infection was associated with increased mortality. In doing so, it highlights the need for nurses to immediately instigate contact precautions for persons suspected of having Clostridium difficile infection and to facilitate a timely faecal collection for testing. Our findings support ongoing surveillance of Clostridium difficile infection and associated prevention and control activities. © 2013 Blackwell Publishing Ltd.

  14. The pangenome of the genus Clostridium.

    Science.gov (United States)

    Udaondo, Zulema; Duque, Estrella; Ramos, Juan-Luis

    2017-07-01

    The pangenome for the genus Clostridium sensu stricto, which was obtained using highly curated and annotated genomes from 16 species is presented; some of these cause disease, while others are used for the production of added-value chemicals. Multilocus sequencing analysis revealed that species of this genus group into at least two clades that include non-pathogenic and pathogenic strains, suggesting that pathogenicity is dispersed across the phylogenetic tree. The core genome of the genus includes 546 protein families, which mainly comprise those involved in protein translation and DNA repair. The GS-GOGAT may represent the central pathway for generating organic nitrogen from inorganic nitrogen sources. Glycerol and glucose metabolism genes are well represented in the core genome together with a set of energy conservation systems. A metabolic network comprising proteins/enzymes, RNAs and metabolites, whose topological structure is a non-random and scale-free network with hierarchically structured modules was built. These modules shed light on the interactions between RNAs, proteins and metabolites, revealing biological features of transcription and translation, cell wall biosynthesis, C1 metabolism and N metabolism. Network analysis identified four nodes that function as hubs and bottlenecks, namely, coenzyme A, HPr kinases, S-adenosylmethionine and the ribonuclease P-protein, suggesting pivotal roles for them in Clostridium. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  15. Secretion of clostridium cellulase by E. coli

    Science.gov (United States)

    Yu, Ida Kuo

    1998-01-01

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

  16. Key Research Issues in Clostridium difficile

    Directory of Open Access Journals (Sweden)

    George Zhanel

    2005-01-01

    Full Text Available 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.

  17. New industrial butanol-producing organism, Clostridium amylovorum

    Energy Technology Data Exchange (ETDEWEB)

    Cataldi, M S

    1964-01-01

    A new Clostridium was isolated from starch-containing substances; it ferments corn and potato starch and sugar molasses, giving important yields of butanol and acetone; it is gram-positive, strictly anaerobic and sporulates in plectron form.

  18. Clostridium difficile infection in the community: a zoonotic disease?

    NARCIS (Netherlands)

    Hensgens, M.P.; Keessen, E.C.; Squire, M.M.; Riley, T.V.; Koene, M.G.J.; de Boer, E.; Lipman, L.J.A.; Kuijper, E.J.

    2012-01-01

    Clostridium difficile infections (CDIs) are traditionally seen in elderly and hospitalized patients who have used antibiotic therapy. In the community, CDIs requiring a visit to a general practitioner are increasingly occurring among young and relatively healthy individuals without known

  19. Clostridium difficile infections in the community: a zoonotic disease?

    NARCIS (Netherlands)

    Hensgens, M.P.M.; Keessen, A.M.; Squire, M.M.; Riley, T.V.; Koene, M.G.J.; Boer, de E.; Lipman, L.J.; Kuijper, E.J.

    2012-01-01

    Clostridium difficile infections (CDIs) are traditionally seen in elderly and hospitalized patients who have used antibiotic therapy. In the community, CDIs requiring a visit to a general practitioner are increasingly occurring among young and relatively healthy individuals without known

  20. Comparison of media for enumeration of Clostridium perfringens from foods

    NARCIS (Netherlands)

    Jong, A.E.I. de; Eijhusen, G.P.; Brouwer-Post, E.J.F.; Grand, M.; Johansson, T.; Kärkkäinen, T.; Marugg, J.; Veld, P.H. in 't; Warmerdam, F.H.M.; Wörner, G.; Zicavo, A.; Rombouts, F.M.; Beumer, R.R.

    2003-01-01

    Many media have been developed to enumerate Clostridium perfringens from foods. In this study, six media [iron sulfite (IS) agar, tryptose sulfite cycloserine (TSC) agar, Shahidi Ferguson perfringens (SFP) agar, sulfite cycloserine azide (SCA), differential clostridial agar (DCA), and oleandomycin

  1. effluent by bacillus cereus and clostridium butyricum using

    African Journals Online (AJOL)

    user

    Double-chambered MFCs was used for the study and operated ..... The third one is wire electron transfer, which uses ... phase indicates that the Bacillus cereus and Clostridium butyricum ..... Improving Start Up Performance With Carbon Mesh.

  2. Clostridium difficile Infection Worsens the Prognosis of Ulcerative Colitis

    Directory of Open Access Journals (Sweden)

    María E Negrón

    2014-01-01

    Full Text Available BACKGROUND: The impact of Clostridium difficile infections among ulcerative colitis (UC patients is well characterized. However, there is little knowledge regarding the association between C difficile infections and postoperative complications among UC patients.

  3. Clostridium difficile infection in Europe: a hospital-based survey

    DEFF Research Database (Denmark)

    Bauer, Martijn P; Notermans, Daan W; van Benthem, Birgit H B

    2011-01-01

    Little is known about the extent of Clostridium difficile infection in Europe. Our aim was to obtain a more complete overview of C difficile infection in Europe and build capacity for diagnosis and surveillance.......Little is known about the extent of Clostridium difficile infection in Europe. Our aim was to obtain a more complete overview of C difficile infection in Europe and build capacity for diagnosis and surveillance....

  4. Reactive arthritis induced by recurrent Clostridium difficile colitis

    Directory of Open Access Journals (Sweden)

    Allison Marr

    2012-01-01

    Full Text Available Clostridium difficile colitis is a common infection that can be difficult to resolve and may result in recurrent infections. Reactive arthritis is a rare presentation of this disease and its treatment is not well differentiated in the literature. We describe a case of reactive arthritis occurring in a patient with a history of recurrent Clostridium difficile colitis while currently receiving a taper of oral vancomycin. His arthritis symptoms resolved with corticosteroids and continued treatment with anticlostridial antibiotics.

  5. Updates on the sporulation process in Clostridium species.

    Science.gov (United States)

    Talukdar, Prabhat K; Olguín-Araneda, Valeria; Alnoman, Maryam; Paredes-Sabja, Daniel; Sarker, Mahfuzur R

    2015-05-01

    Sporulation is an important strategy for certain bacterial species within the phylum Firmicutes to survive longer periods of time in adverse conditions. All spore-forming bacteria have two phases in their life; the vegetative form, where they can maintain all metabolic activities and replicate to increase numbers, and the spore form, where no metabolic activities exist. Although many essential components of sporulation are conserved among the spore-forming bacteria, there are differences in the regulation and the pathways among different genera, even at the species level. While we have gained much information from the most studied spore-forming bacterial genus, Bacillus, we still lack an in-depth understanding of spore formation in the genus Clostridium. Clostridium and Bacillus share the master regulator of sporulation, Spo0A, and its downstream pathways, but there are differences in the activation of the Spo0A pathway. While Bacillus species use a multi-component phosphorylation pathway for phosphorylation of Spo0A, termed phosphorelay, such a phosphorelay system is absent in Clostridium. On the other hand, a number of genes regulated by the different sporulation-specific transcription factors are conserved between different Clostridium and Bacillus species. In this review, we discuss the recent findings on Clostridium sporulation and compare the sporulation mechanism in Clostridium and Bacillus. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  6. Biosynthesis of dipicolinic acid in Clostridium roseum

    Energy Technology Data Exchange (ETDEWEB)

    Prakasan, K. (Paraiba Univ., Joao Pessoa (Brazil)); Sharma, D. (Gobind Ballabh Pant Univ. of Agriculture and Technology, Nainital (India))

    1981-02-01

    Dipicolinic acid (DPA) synthesis was studied in Clostridium roseum by permitting the organism to complete vegetative growth in trypticase medium and trasfering the cells to a non-growth-promoting-medium, supplemented with the appropriate /sup 14/C-labelled precursors to complete sporulation and assaying the incorporation of label into DPA. Glu, asp, ala, ser and acetate were found to be efficient precursors of DPA and each one influenced the incorporation of other into DPA. The data suggest that a C/sub 5/ precursor is being trasformed into a C/sub 4/ intermediate, and a C/sub 2/ precursor into a C/sub 4/ intermediate, before their entry into DPA carbon structure. A C/sub 4/ plus C/sub 3/ condensation is favoured over C/sub 5/ plus C/sub 2/ or other condensation in the DPA biosynthesis.

  7. Biosynthesis of dipicolinic acid in Clostridium roseum

    International Nuclear Information System (INIS)

    Prakasan, K.; Sharma, D.

    1981-01-01

    Dipicolinic acid (DPA) synthesis was studied in Clostridium roseum by permitting the organism to complete vegetative growth in trypticase medium and trasfering the cells to a non-growth-promoting-medium, supplemented with the appropriate 14 C-labelled precursors to complete sporulation and assaying the incorporation of label into DPA. Glu, asp, ala, ser and acetate were found to be efficient precursors of DPA and each one influenced the incorporation of other into DPA. The data suggest that a C 5 precursor is being trasformed into a C 4 intermediate, and a C 2 precursor into a C 4 intermediate, before their entry into DPA carbon structure. A C 4 plus C 3 condensation is favoured over C 5 plus C 2 or other condensation in the DPA biosynthesis. (Author) [pt

  8. Handling Gene and Protein Names in the Age of Bioinformatics: The Special Challenge of Secreted Multimodular Bacterial Enzymes such as the cbhA/cbh9A Gene of Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Brunecky, Roman [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Schwarz, Wolfgang H. [Technical University of Munich; Broeker, Jannis [Technical University of Munich; Liebl, Wolfgang [Technical University of Munich; Zverlov, Vladimir V. [Technical University of Munich; Russian Academy of Science

    2018-02-26

    An increasing number of researchers working in biology, biochemistry, biotechnology, bioengineering, bioinformatics and other related fields of science are using biological molecules. As the scientific background of the members of different scientific communities is more diverse than ever before, the number of scientists not familiar with the rules for non-ambiguous designation of genetic elements is increasing. However, with biological molecules gaining importance through biotechnology, their functional and unambiguous designation is vital. Unfortunately, naming genes and proteins is not an easy task. In addition, the traditional concepts of bioinformatics are challenged with the appearance of proteins comprising different modules with a respective function in each module. This article highlights basic rules and novel solutions in designation recently used within the community of bacterial geneticists, and we discuss the present-day handling of gene and protein designations. As an example we will utilize a recent mischaracterization of gene nomenclature. We make suggestions for better handling of names in future literature as well as in databases and annotation projects. Our methodology emphasizes the hydrolytic function of multi-modular genes and extracellular proteins from bacteria.

  9. Viability of Clostridium sporogenes spores after CaO hygienization of meat waste

    Directory of Open Access Journals (Sweden)

    Justyna Bauza-Kaszewska

    2014-09-01

    Full Text Available The occurrence of the pathogenic species [i]C. perfringens[/i] and [i]C. botulinum spores[/i] in animal by-products poses a potential epidemiological hazard. Strong entero- and neurotoxins produced by these bacteria adversely affect human health. To inactivate pathogens present in animal by-products, waste must be subjected to various methods of sanitization. The aim of the presented study was to estimate the effect of different doses of CaO on the viability of spores [i] Clostridium sporogenes[/i] in meat wastes category 3. During the research, two doses of burnt lime were added to the poultry mince meat and meat mixed with swine blood contaminated with [i]Clostridium sporogenes[/i] spore suspension. Half of the samples collected for microbiological analyses were buffered to achieve the pH level ~7, the other were examined without pH neutralization. To estimate the spore number, 10-fold dilution series in peptone water was prepared and heat-treated at 80 °C for 10 min. After cooling-down, one milliliter of each dilution was pour-plated onto DRCM medium solidified with agar. Statistical analysis were performed using the Statistica software. Application of 70% CaO caused complete inactivation of [i]Clostridium spores[/i] in meat wastes after 48 hours. The highest temperature achieved during the experiment was 67 °C. Rapid alkalization of the biomass resulted in increasing pH to values exceeding 12. The effect of liming was not dependent on the meat wastes composition nor CaO dose. The experiment proved the efficiency of liming as a method of animal by-products sanitization. Application of the obtained results may help reduce the epidemiological risk and ensure safety to people handling meat wastes at each stage of their processing and utilization.

  10. NCBI nr-aa BLAST: CBRC-RNOR-05-0228 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-05-0228 ref|YP_001038304.1| cellulosome enzyme, dockerin type I [Clostrid...ium thermocellum ATCC 27405] gb|ABN53111.1| cellulosome enzyme, dockerin type I [Clostridium thermocellum ATCC 27405] YP_001038304.1 5e-07 31% ...

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

    Science.gov (United States)

    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 perfringens isolate typing by multiplex PCR

    Directory of Open Access Journals (Sweden)

    MR Ahsani

    2010-01-01

    Full Text Available Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39% type A, 21.74% type B, 34.78% type C and 26.09% type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.

  13. The economic burden of Clostridium difficile

    Science.gov (United States)

    McGlone, S. M.; Bailey, R. R.; Zimmer, S. M.; Popovich, M. J.; Tian, Y.; Ufberg, P.; Muder, R. R.; Lee, B. Y.

    2013-01-01

    Although Clostridium difficile (C. difficile) is the leading cause of infectious diarrhoea in hospitalized patients, the economic burden of this major nosocomial pathogen for hospitals, third-party payers and society remains unclear. We developed an economic computer simulation model to determine the costs attributable to healthcare-acquired C. difficile infection (CDI) from the hospital, third-party payer and societal perspectives. Sensitivity analyses explored the effects of varying the cost of hospitalization, C. difficile-attributable length of stay, and the probability of initial and secondary recurrences. The median cost of a case ranged from $9179 to $11 456 from the hospital perspective, $8932 to $11 679 from the third-party payor perspective, and $13 310 to $16 464 from the societal perspective. Most of the costs incurred were accrued during a patient’s primary CDI episode. Hospitals with an incidence of 4.1 CDI cases per 100 000 discharges would incur costs ≥$3.2 million (hospital perspective); an incidence of 10.5 would lead to costs ≥$30.6 million. Our model suggests that the annual US economic burden of CDI would be ≥$496 million (hospital perspective), ≥$547 million (third-party payer perspective) and ≥$796 million (societal perspective). Our results show that C. difficile infection is indeed costly, not only to third-party payers and the hospital, but to society as well. These results are consistent with current literature citing C. difficile as a costly disease. PMID:21668576

  14. CLOSTRIDIUM PERFRINGENS IN MEAT AND MEAT PRODUCTS.

    Science.gov (United States)

    HALL, H E; ANGELOTTI, R

    1965-05-01

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

  15. Clostridium difficile is an autotrophic bacterial pathogen.

    Directory of Open Access Journals (Sweden)

    Michael Köpke

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

  16. Phosphorylation of proteins in Clostridium thermohydrosulfuricum

    International Nuclear Information System (INIS)

    Londesborough, J.

    1986-01-01

    Cell extracts of the thermophile Clostridium thermohydrosulfuricum catalyzed the phosphorylation by (γ- 32 P)ATP of several endogenous proteins with M/sub r/s between 13,000 and 100,000. Serine and tyrosine were the main acceptors. Distinct substrate proteins were found in the soluble (e.g., proteins p66, p63, and p53 of M/sub r/s 66,000, 63,000, and 53,000, respectively) and particulate (p76 and p30) fractions, both of which contained protein kinase and phosphatase activity. The soluble fraction suppressed the phosphorylation of particulate proteins and contained a protein kinase inhibitor. Phosphorylation of p53 was promoted by 10μM fructose 1,6-bisphosphate or glucose 1,6-bisphosphate and suppressed by hexose monophosphates, whereas p30 and p13 were suppressed by 5 μM brain (but not spinach) calmodulin. Polyamines, including the odd polyamines characteristic of thermophiles, modulated the labeling of most of the phosphoproteins. Apart from p66, all the proteins labeled in vitro were also rapidly labeled in intact cells by 32 P/sub i/. Several proteins strongly labeled in vivo were labeled slowly or not at all in vitro

  17. Crystal structure of Clostridium difficile toxin A

    Energy Technology Data Exchange (ETDEWEB)

    Chumbler, Nicole M.; Rutherford, Stacey A.; Zhang, Zhifen; Farrow, Melissa A.; Lisher, John P.; Farquhar, Erik; Giedroc, David P.; Spiller, Benjamin W.; Melnyk, Roman A.; Lacy, D. Borden

    2016-01-11

    Clostridium difficile infection is the leading cause of hospital-acquired diarrhoea and pseudomembranous colitis. Disease is mediated by the actions of two toxins, TcdA and TcdB, which cause the diarrhoea, as well as inflammation and necrosis within the colon. The toxins are large (308 and 270 kDa, respectively), homologous (47% amino acid identity) glucosyltransferases that target small GTPases within the host. The multidomain toxins enter cells by receptor-mediated endocytosis and, upon exposure to the low pH of the endosome, insert into and deliver two enzymatic domains across the membrane. Eukaryotic inositol-hexakisphosphate (InsP6) binds an autoprocessing domain to activate a proteolysis event that releases the N-terminal glucosyltransferase domain into the cytosol. Here, we report the crystal structure of a 1,832-amino-acid fragment of TcdA (TcdA1832), which reveals a requirement for zinc in the mechanism of toxin autoprocessing and an extended delivery domain that serves as a scaffold for the hydrophobic α-helices involved in pH-dependent pore formation. A surface loop of the delivery domain whose sequence is strictly conserved among all large clostridial toxins is shown to be functionally important, and is highlighted for future efforts in the development of vaccines and novel therapeutics.

  18. Clostridium difficile – From Colonization to Infection

    Science.gov (United States)

    Schäffler, Holger; Breitrück, Anne

    2018-01-01

    Clostridium difficile is the most frequent cause of nosocomial antibiotic-associated diarrhea. The incidence of C. difficile infection (CDI) has been rising worldwide with subsequent increases in morbidity, mortality, and health care costs. Asymptomatic colonization with C. difficile is common and a high prevalence has been found in specific cohorts, e.g., hospitalized patients, adults in nursing homes and in infants. However, the risk of infection with C. difficile differs significantly between these cohorts. While CDI is a clear indication for therapy, colonization with C. difficile is not believed to be a direct precursor for CDI and therefore does not require treatment. Antibiotic therapy causes alterations of the intestinal microbial composition, enabling C. difficile colonization and consecutive toxin production leading to disruption of the colonic epithelial cells. Clinical symptoms of CDI range from mild diarrhea to potentially life-threatening conditions like pseudomembranous colitis or toxic megacolon. While antibiotics are still the treatment of choice for CDI, new therapies have emerged in recent years such as antibodies against C. difficile toxin B and fecal microbial transfer (FMT). This specific therapy for CDI underscores the role of the indigenous bacterial composition in the prevention of the disease in healthy individuals and its role in the pathogenesis after alteration by antibiotic treatment. In addition to the pathogenesis of CDI, this review focuses on the colonization of C. difficile in the human gut and factors promoting CDI. PMID:29692762

  19. Emerging monoclonal antibodies against Clostridium difficile infection.

    Science.gov (United States)

    Péchiné, Séverine; Janoir, Claire; Collignon, Anne

    2017-04-01

    Clostridium difficile infections are characterized by a high recurrence rate despite antibiotic treatments and there is an urgent need to develop new treatments such as fecal transplantation and immonotherapy. Besides active immunotherapy with vaccines, passive immunotherapy has shown promise, especially with monoclonal antibodies. Areas covered: Herein, the authors review the different assays performed with monoclonal antibodies against C. difficile toxins and surface proteins to treat or prevent primary or recurrent episodes of C. difficile infection in animal models and in clinical trials as well. Notably, the authors lay emphasis on the phase III clinical trial (MODIFY II), which allowed bezlotoxumab to be approved by the Food and Drug Administration and the European Medicines Agency. They also review new strategies for producing single domain antibodies and nanobodies against C. difficile and new approaches to deliver them in the digestive tract. Expert opinion: Only two human Mabs against TcdA and TcdB have been tested alone or in combination in clinical trials. However, many animal model studies have provided rationale for the use of Mabs and nanobodies in C. difficile infection and pave the way for further clinical investigation.

  20. [Epidemiology of Clostridium difficile infection in Spain].

    Science.gov (United States)

    Asensio, Angel; Monge, Diana

    2012-06-01

    There has been increasing interest in Clostridium difficile infection (CDI) due its association with healthcare and its impact on morbidity and mortality in the elderly. During the last few years there has been a growing increase in the number of published studies on the incidence, changes on the clinical presentation and on the epidemiology, with the description of new risk factors. The frequency of CDI in Spain is not sufficiently characterised. The available data indicates that incidence is within the range of that of surrounding countries but increasing. Furthermore, the high and growing use of broad spectrum antibiotics, both in our hospitals and in the community setting, are factors that favour the increase of the disease. The hyper-virulent ribotype 027 has not spread in our hospitals. We need to know with enhanced validity and accuracy the incidence of CDI, both community and healthcare-associated, the information on outbreaks, the incidence on certain population groups, the characterisation of circulating ribotypes and the impact of the disease in terms of mortality and health costs. We need to implement programs for the improvement of antibiotic therapy in the hospital, as well as in the community. Furthermore, the knowledge and the performance of standard precautions need to be improved, particularly hand hygiene, and the specific measures to limit the transmission of C. difficile among the healthcare institutions. Copyright © 2011 Elsevier España, S.L. All rights reserved.

  1. Antibiotic prescribing policy and Clostridium difficile diarrhoea.

    LENUS (Irish Health Repository)

    O'Connor, K A

    2012-02-03

    BACKGROUND: Broad-spectrum antibiotics, particularly intravenous cephalosporins, are associated with Clostridium difficile diarrhoea. Diarrhoea due to C. difficile is a growing problem in hospitals, especially among elderly patients. AIM: To establish whether changing an antibiotic policy with the aim of reducing the use of injectable cephalosporins leads to a reduction in the incidence of C. difficile diarrhoea in elderly patients. DESIGN: Retrospective analysis. METHODS: A group of patients who were subject to the new antibiotic policy from the period following July 2000, were compared with patients who were admitted prior to July 2000 and were not subject to the new policy. Infections, antibiotic prescriptions and mortality rates were determined from case notes, and C. difficle diarrhoea rates from microbiological data. RESULTS: Intravenous cephalosporin use fell from 210 to 28 defined daily doses (p < 0.001) following the change in antibiotic policy, with a corresponding increase in piperacillin-tazobactam (p < 0.001) and moxifloxacin (p < 0.001) use. The new policy led to a significant reduction in C. difficile diarrhoea cases. The relative risk of developing C. difficile infection with the old policy compared to the new policy was 3.24 (95%CI 1.07-9.84, p = 0.03). DISCUSSION: The antibiotic policy was successfully introduced into an elderly care service. It reduced both intravenous cephalosporin use and C. difficile diarrhoea.

  2. Sepsis due to clostridium septicum: case report

    International Nuclear Information System (INIS)

    Foga, M.M.; McGinn, G.J.; Kroeker, M.A.; Guzman, R.

    2000-01-01

    Clostridium septicum is an unusual anaerobic, gram-positive, gas-producing bacillus, which has been identified as a cause of fulminant rapidly fatal infection in humans. Infection with C. septicum usually occurs in patients with cancer, patients receiving immunosuppressive chemotherapy, or patients with a nonmalignant hematological disorder such as hemolytic uremic syndrome. C. septicum infection most commonly involves the abdomen, and a recent review article has identified 164 cases in the medical literature describing the abdominal findings in this disease. Intracranial manifestation of C. septicum infection are less common and include meningitis, cerebritis, abscess formation and pneumocephalus. There have been only 12 documented cases in the English literature describing central nervous system lesions associated with C. septicum. We present a case report of a 56-year-old man in whom septicemia due to C. septicum developed as a complication of Crohn's disease. To our knowledge, there has never been a previous report of C. septicum sepsis related to underlying Crohn's disease. Our case is also remarkable in that an intracerebral gas collection developed at the site of a mycotic infarct related to C. septicum bacteremia, Intracranial, intraparenchymal gas formation related to anaerobic infection is extremely rare; to our knowledge, this radiological finding related to C. septicum sepsis has been described in only 1 previous case report in the medical literature. We also describe the intra-abdominal manifestations of C. septicum sepsis that occurred in this patient as well as the associated radiographic and pathologic findings. (author)

  3. Perfringolysin O: The Underrated Clostridium perfringens Toxin?

    Science.gov (United States)

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

    2015-05-14

    The anaerobic bacterium Clostridium perfringens expresses multiple toxins that promote disease development in both humans and animals. One such toxin is perfringolysin O (PFO, classically referred to as θ toxin), a pore-forming cholesterol-dependent cytolysin (CDC). PFO is secreted as a water-soluble monomer that recognizes and binds membranes via cholesterol. Membrane-bound monomers undergo structural changes that culminate in the formation of an oligomerized prepore complex on the membrane surface. The prepore then undergoes conversion into the bilayer-spanning pore measuring approximately 250-300 Å in diameter. PFO is expressed in nearly all identified C. perfringens strains and harbors interesting traits that suggest a potential undefined role for PFO in disease development. Research has demonstrated a role for PFO in gas gangrene progression and bovine necrohemorrhagic enteritis, but there is limited data available to determine if PFO also functions in additional disease presentations caused by C. perfringens. This review summarizes the known structural and functional characteristics of PFO, while highlighting recent insights into the potential contributions of PFO to disease pathogenesis.

  4. Perfringolysin O: The Underrated Clostridium perfringens Toxin?

    Directory of Open Access Journals (Sweden)

    Stefanie Verherstraeten

    2015-05-01

    Full Text Available The anaerobic bacterium Clostridium perfringens expresses multiple toxins that promote disease development in both humans and animals. One such toxin is perfringolysin O (PFO, classically referred to as θ toxin, a pore-forming cholesterol-dependent cytolysin (CDC. PFO is secreted as a water-soluble monomer that recognizes and binds membranes via cholesterol. Membrane-bound monomers undergo structural changes that culminate in the formation of an oligomerized prepore complex on the membrane surface. The prepore then undergoes conversion into the bilayer-spanning pore measuring approximately 250–300 Å in diameter. PFO is expressed in nearly all identified C. perfringens strains and harbors interesting traits that suggest a potential undefined role for PFO in disease development. Research has demonstrated a role for PFO in gas gangrene progression and bovine necrohemorrhagic enteritis, but there is limited data available to determine if PFO also functions in additional disease presentations caused by C. perfringens. This review summarizes the known structural and functional characteristics of PFO, while highlighting recent insights into the potential contributions of PFO to disease pathogenesis.

  5. The Clostridium sporulation programs: diversity and preservation of endospore differentiation.

    Science.gov (United States)

    Al-Hinai, Mohab A; Jones, Shawn W; Papoutsakis, Eleftherios T

    2015-03-01

    Bacillus and Clostridium organisms initiate the sporulation process when unfavorable conditions are detected. The sporulation process is a carefully orchestrated cascade of events at both the transcriptional and posttranslational levels involving a multitude of sigma factors, transcription factors, proteases, and phosphatases. Like Bacillus genomes, sequenced Clostridium genomes contain genes for all major sporulation-specific transcription and sigma factors (spo0A, sigH, sigF, sigE, sigG, and sigK) that orchestrate the sporulation program. However, recent studies have shown that there are substantial differences in the sporulation programs between the two genera as well as among different Clostridium species. First, in the absence of a Bacillus-like phosphorelay system, activation of Spo0A in Clostridium organisms is carried out by a number of orphan histidine kinases. Second, downstream of Spo0A, the transcriptional and posttranslational regulation of the canonical set of four sporulation-specific sigma factors (σ(F), σ(E), σ(G), and σ(K)) display different patterns, not only compared to Bacillus but also among Clostridium organisms. Finally, recent studies demonstrated that σ(K), the last sigma factor to be activated according to the Bacillus subtilis model, is involved in the very early stages of sporulation in Clostridium acetobutylicum, C. perfringens, and C. botulinum as well as in the very late stages of spore maturation in C. acetobutylicum. Despite profound differences in initiation, propagation, and orchestration of expression of spore morphogenetic components, these findings demonstrate not only the robustness of the endospore sporulation program but also the plasticity of the program to generate different complex phenotypes, some apparently regulated at the epigenetic level. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  6. The Rise and Fall of Metronidazole for Clostridium difficile Infection.

    Science.gov (United States)

    Chahine, Elias B

    2018-06-01

    Clostridium difficile is posing urgent health threats. Older studies have shown that metronidazole and vancomycin are equally effective in the treatment of Clostridium difficile infection (CDI). Given its inexpensive cost and low propensity to select antimicrobial resistant organisms, metronidazole became rapidly the drug of choice despite its pharmacokinetic limitations in the treatment of CDI. However, newer studies demonstrated that metronidazole is inferior to vancomycin, prompting clinicians to change their long-standing position on using metronidazole for mild to moderate infections and on reserving vancomycin for severe infections. Moving forward, metronidazole will fall out of favor in the treatment of CDI.

  7. Pleiotropic roles of Clostridium difficile sin locus

    Science.gov (United States)

    Ou, Junjun; Dupuy, Bruno

    2018-01-01

    Clostridium difficile is the primary cause of nosocomial diarrhea and pseudomembranous colitis. It produces dormant spores, which serve as an infectious vehicle responsible for transmission of the disease and persistence of the organism in the environment. In Bacillus subtilis, the sin locus coding SinR (113 aa) and SinI (57 aa) is responsible for sporulation inhibition. In B. subtilis, SinR mainly acts as a repressor of its target genes to control sporulation, biofilm formation, and autolysis. SinI is an inhibitor of SinR, so their interaction determines whether SinR can inhibit its target gene expression. The C. difficile genome carries two sinR homologs in the operon that we named sinR and sinR’, coding for SinR (112 aa) and SinR’ (105 aa), respectively. In this study, we constructed and characterized sin locus mutants in two different C. difficile strains R20291 and JIR8094, to decipher the locus’s role in C. difficile physiology. Transcriptome analysis of the sinRR’ mutants revealed their pleiotropic roles in controlling several pathways including sporulation, toxin production, and motility in C. difficile. Through various genetic and biochemical experiments, we have shown that SinR can regulate transcription of key regulators in these pathways, which includes sigD, spo0A, and codY. We have found that SinR’ acts as an antagonist to SinR by blocking its repressor activity. Using a hamster model, we have also demonstrated that the sin locus is needed for successful C. difficile infection. This study reveals the sin locus as a central link that connects the gene regulatory networks of sporulation, toxin production, and motility; three key pathways that are important for C. difficile pathogenesis. PMID:29529083

  8. CRISPR Diversity and Microevolution in Clostridium difficile.

    Science.gov (United States)

    Andersen, Joakim M; Shoup, Madelyn; Robinson, Cathy; Britton, Robert; Olsen, Katharina E P; Barrangou, Rodolphe

    2016-09-19

    Virulent strains of Clostridium difficile have become a global health problem associated with morbidity and mortality. Traditional typing methods do not provide ideal resolution to track outbreak strains, ascertain genetic diversity between isolates, or monitor the phylogeny of this species on a global basis. Here, we investigate the occurrence and diversity of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) in C. difficile to assess the potential of CRISPR-based phylogeny and high-resolution genotyping. A single Type-IB CRISPR-Cas system was identified in 217 analyzed genomes with cas gene clusters present at conserved chromosomal locations, suggesting vertical evolution of the system, assessing a total of 1,865 CRISPR arrays. The CRISPR arrays, markedly enriched (8.5 arrays/genome) compared with other species, occur both at conserved and variable locations across strains, and thus provide a basis for typing based on locus occurrence and spacer polymorphism. Clustering of strains by array composition correlated with sequence type (ST) analysis. Spacer content and polymorphism within conserved CRISPR arrays revealed phylogenetic relationship across clades and within ST. Spacer polymorphisms of conserved arrays were instrumental for differentiating closely related strains, e.g., ST1/RT027/B1 strains and pathogenicity locus encoding ST3/RT001 strains. CRISPR spacers showed sequence similarity to phage sequences, which is consistent with the native role of CRISPR-Cas as adaptive immune systems in bacteria. Overall, CRISPR-Cas sequences constitute a valuable basis for genotyping of C. difficile isolates, provide insights into the micro-evolutionary events that occur between closely related strains, and reflect the evolutionary trajectory of these genomes. © The Author(s) 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  9. CRISPR Diversity and Microevolution in Clostridium difficile

    Science.gov (United States)

    Andersen, Joakim M.; Shoup, Madelyn; Robinson, Cathy; Britton, Robert; Olsen, Katharina E.P.; Barrangou, Rodolphe

    2016-01-01

    Abstract Virulent strains of Clostridium difficile have become a global health problem associated with morbidity and mortality. Traditional typing methods do not provide ideal resolution to track outbreak strains, ascertain genetic diversity between isolates, or monitor the phylogeny of this species on a global basis. Here, we investigate the occurrence and diversity of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) in C. difficile to assess the potential of CRISPR-based phylogeny and high-resolution genotyping. A single Type-IB CRISPR-Cas system was identified in 217 analyzed genomes with cas gene clusters present at conserved chromosomal locations, suggesting vertical evolution of the system, assessing a total of 1,865 CRISPR arrays. The CRISPR arrays, markedly enriched (8.5 arrays/genome) compared with other species, occur both at conserved and variable locations across strains, and thus provide a basis for typing based on locus occurrence and spacer polymorphism. Clustering of strains by array composition correlated with sequence type (ST) analysis. Spacer content and polymorphism within conserved CRISPR arrays revealed phylogenetic relationship across clades and within ST. Spacer polymorphisms of conserved arrays were instrumental for differentiating closely related strains, e.g., ST1/RT027/B1 strains and pathogenicity locus encoding ST3/RT001 strains. CRISPR spacers showed sequence similarity to phage sequences, which is consistent with the native role of CRISPR-Cas as adaptive immune systems in bacteria. Overall, CRISPR-Cas sequences constitute a valuable basis for genotyping of C. difficile isolates, provide insights into the micro-evolutionary events that occur between closely related strains, and reflect the evolutionary trajectory of these genomes. PMID:27576538

  10. Effect of a probiotic on prevention of diarrhea and Clostridium difficile and Clostridium perfringens shedding in foals

    DEFF Research Database (Denmark)

    Schoster, Angelika; Staempfli, H R; Abrahams, M

    2015-01-01

    of incidence and duration of diarrhea and fecal shedding of Clostridium perfringens and Clostridium difficile between treatment and age groups. RESULTS: The overall incidence of diarrhea was 41 of 72 (59%) and did not differ (P = 0.37) between treatment groups. Foals treated with probiotics were more likely...... of C. perfringens shedding was 55% with no difference between treatment groups (P = 0.23). The prevalence of C. difficile shedding was 11%. CONCLUSION AND CLINICAL IMPORTANCE: There was no benefit of administering a 3-week course of probiotics, but potential adverse effects were noted. Whether...

  11. [Detailed methodological recommendations for the treatment of Clostridium difficile-associated diarrhea with faecal transplantation].

    Science.gov (United States)

    Nagy, Gergely György; Várvölgyi, Csaba; Balogh, Zoltán; Orosi, Piroska; Paragh, György

    2013-01-06

    The incidence of Clostridium difficile associated enteral disease shows dramatic increase worldwide, with appallingly high treatment costs, mortality figures, recurrence rates and treatment refractoriness. It is not surprising, that there is significant interest in the development and introduction of alternative therapeutic strategies. Among these only stool transplantation (or faecal bacteriotherapy) is gaining international acceptance due to its excellent cure rate (≈92%), low recurrence rate (≈6%), safety and cost-effectiveness. Unfortunately faecal transplantation is not available for most patients, although based on promising international results, its introduction into the routine clinical practice is well justified and widely expected. The authors would like to facilitate this process, by presenting a detailed faecal transplantation protocol prepared in their Institution based on the available literature and clinical rationality. Officially accepted national methodological guidelines will need to be issued in the future, founded on the expert opinion of relevant professional societies and upcoming advances in this field.

  12. Cloning in Escherichia coli of the enterotoxin gene from Clostridium perfringens type A.

    Science.gov (United States)

    Iwanejko, L A; Routledge, M N; Stewart, G S

    1989-04-01

    A 26 bp DNA probe has been constructed with minimal degeneracy to the protein sequence for Clostridium perfringens enterotoxin. The probe has been hybridized against a 6-10 kb chromosomal bank from C. perfringens 8239, prepared as a HindIII partial digest in pHG165. From this survey a clone has been identified containing a 6.8 kb DNA insert with strong hybridization to the probe. Direct plasmid sequencing has identified a translational reading frame within this clone which correlates with the known protein sequence for the type A enterotoxin. DNA sequences 5' to this open reading frame and containing the putative transcriptional control regions show areas of significant homology with regions upstream from the ATG codon of the tetanus toxin gene.

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

    Science.gov (United States)

    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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Uterine Clostridium perfringens infection related to gynecologic malignancy.

    Science.gov (United States)

    Kremer, Kevin M; McDonald, Megan E; Goodheart, Michael J

    2017-11-01

    Uterine gas gangrene caused by Clostridium perfringens is a serious, often life-threatening infection that is rarely encountered in the practice of gynecologic oncology. However, the hypoxic nature of gynecologic cancers due to necrosis and/or prior radiation therapy creates a microenvironment optimal for proliferation of anaerobic bacteria such as the Clostridium species. Early recognition and aggressive treatment with IV antibiotics and surgical debridement remain the cornerstones of management in order to decrease morbidity and mortality. Here we present the case of a 52 year-old woman with a remote history of cervical cancer who was previously treated at our institution with primary chemotherapy and radiation and was then admitted decades later with Clostridium perfringens bacteremia and CT evidence of intrauterine abscess. The patient received a prolonged course of IV antibiotic therapy and subsequently underwent definitive surgical management with a total abdominal hysterectomy, bilateral salpingo-oophorectomy, small bowel resection with anastomosis for a utero-ileal fistula identified intraoperatively. Pathology from the uterine specimen demonstrated a primary poorly differentiated uterine adenocarcinoma. The patient recovered fully from her Clostridium perfringens infection and was discharged from the hospital shortly after surgical intervention.

  15. Clostridium difficile in piglets in the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Goldová, Jana; Malinová, A.; Indra, A.; Vítek, L.; Branny, Pavel; Jirásková, Alena

    2012-01-01

    Roč. 57, č. 2 (2012), s. 159-161 ISSN 0015-5632 R&D Projects: GA AV ČR KJB500200901 Institutional research plan: CEZ:AV0Z50200510 Keywords : clostridium * piglets Subject RIV: EE - Microbiology, Virology Impact factor: 0.791, year: 2012

  16. Duodenal infusion of donor feces for recurrent Clostridium difficile

    NARCIS (Netherlands)

    van Nood, Els; Vrieze, Anne; Nieuwdorp, Max; Fuentes, Susana; Zoetendal, Erwin G.; de Vos, Willem M.; Visser, Caroline E.; Kuijper, Ed J.; Bartelsman, Joep F. W. M.; Tijssen, Jan G. P.; Speelman, Peter; Dijkgraaf, Marcel G. W.; Keller, Josbert J.

    2013-01-01

    Recurrent Clostridium difficile infection is difficult to treat, and failure rates for antibiotic therapy are high. We studied the effect of duodenal infusion of donor feces in patients with recurrent C. difficile infection. We randomly assigned patients to receive one of three therapies: an initial

  17. Flooding and Clostridium difficile infection: a case-crossover analysis

    Science.gov (United States)

    Clostridium difficile is a bacterium that can spread by water. It often causes acute gastrointestinal illness in older adults who are hospttalized and/or receiving antibiotics; however, community­ associated infections affecting otherwise healthy individuals have become more comm...

  18. Effect of cooling on Clostridium perfringens in pea soup

    NARCIS (Netherlands)

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

    2004-01-01

    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

  19. Occurrence of Clostridium perfringens in sausages sold in Meknes ...

    African Journals Online (AJOL)

    In Morocco, the consumption of meat products has experienced a sharp increase in recent years despite the presence of pathogenic bacteria due to hygiene failure. The present study was designed to determine the prevalence of Clostridium perfringens in sausages sold in Meknes city (Morocco) and to study the different ...

  20. 9 CFR 113.106 - Clostridium Chauvoei Bacterin.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Clostridium Chauvoei Bacterin. 113.106 Section 113.106 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF... Animal and Plant Health Inspection Service, shall be used for challenge 14 to 15 days following the last...

  1. 9 CFR 113.107 - Clostridium Haemolyticum Bacterin.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Clostridium Haemolyticum Bacterin. 113.107 Section 113.107 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT... challenge 14 to 15 days following the last injection of the product. Each of eight vaccinates and each of...

  2. Cost-effectiveness in Clostridium difficile treatment decision-making

    NARCIS (Netherlands)

    Nuijten, Mark J. C.; Keller, Josbert J.; Visser, Caroline E.; Redekop, Ken; Claassen, Eric; Speelman, Peter; Pronk, Marja H.

    2015-01-01

    To develop a framework for the clinical and health economic assessment for management of Clostridium difficile infection (CDI). CDI has vast economic consequences emphasizing the need for innovative and cost effective solutions, which were aim of this study. A guidance model was developed for

  3. A toxic approach to beta2-toxigenic Clostridium perfringens

    NARCIS (Netherlands)

    Allaart, J.G.

    2013-01-01

    Clostridium perfringens is one of the most important causes of intestinal disease in animals and humans. Its virulence is attributed to the several toxins it can produce, including the beta2 toxin encoded by cpb2. In this thesis we studied the role of the beta2 toxin produced by C. perfringens in

  4. Imipenem Resistance in Clostridium difficile Ribotype 017, Portugal

    Science.gov (United States)

    Isidro, Joana; Santos, Andrea; Nunes, Alexandra; Borges, Vítor; Silva, Catarina; Vieira, Luís; Mendes, Aristides L.; Serrano, Mónica; Henriques, Adriano O.; Gomes, João Paulo

    2018-01-01

    We describe imipenem-resistant and imipenem-susceptible clinical isolates of Clostridium difficile ribotype 017 in Portugal. All ribotype 017 isolates carried an extra penicillin-binding protein gene, pbp5, and the imipenem-resistant isolates had additional substitutions near the transpeptidase active sites of pbp1 and pbp3. These clones could disseminate and contribute to imipenem resistance. PMID:29553322

  5. A Quantitative Electrochemiluminescence Assay for Clostridium perfringens alpha toxin

    Science.gov (United States)

    2006-08-10

    Doyle, L.R. Beuchat, T.J. Montville (Eds.), Food Microbiology : Fundamentals and Fron- tiers, Second ed., ASM Press, Washington, D.C., 2001, pp. 351...D.E. Lorant, A.E. Bryant, G.A. Zimmerman, T.M. McIn- tyre, D.L. Stevens, S.M. Prescott , Alpha toxin from Clostridium per- fringens induces

  6. Fulminant leukemoid reaction due to postpartum Clostridium sordellii infection

    Directory of Open Access Journals (Sweden)

    Prabhat Agrawal

    2012-01-01

    Full Text Available Clostridium sordellii is gram positive anaerobic spore forming rod and it has been demonstrated to cause gas gangrene, refractory shock, leukemoid reaction, and pleuroperitoneal effusion due to capillary leak. We report here a case of postpartum female who presented with leukemoid reaction, ascites, pleural effusion, and shock without fever 7 days after normal vaginal home delivery.

  7. Investigation of Clostridium botulinum group III's mobilome content

    NARCIS (Netherlands)

    Woudstra, Cédric; Maréchal, Le Caroline; Souillard, Rozenn; Anniballi, Fabrizio; Auricchio, Bruna; Bano, Luca; Bayon-Auboyer, Marie Hélène; Koene, Miriam; Mermoud, Isabelle; Brito, Roseane B.; Lobato, Francisco C.F.; Silva, Rodrigo O.S.; Dorner, Martin B.; Fach, Patrick

    2018-01-01

    Clostridium botulinum group III is mainly responsible for botulism in animals. It could lead to high animal mortality rates and, therefore, represents a major environmental and economic concern. Strains of this group harbor the botulinum toxin locus on an unstable bacteriophage. Since the release of

  8. Clostridium difficile infection in an endemic setting in the Netherlands

    NARCIS (Netherlands)

    Hensgens, M. P. M.; Goorhuis, A.; van Kinschot, C. M. J.; Crobach, M. J. T.; Harmanus, C.; Kuijper, E. J.

    2011-01-01

    The purpose of this investigation was to study risk factors for Clostridium difficile infection (CDI) in an endemic setting. In a 34-month prospective case-control study, we compared the risk factors and clinical characteristics of all consecutively diagnosed hospitalised CDI patients (n = 93) with

  9. The morbidity, mortality, and costs associated with Clostridium difficile infection.

    Science.gov (United States)

    Kwon, Jennie H; Olsen, Margaret A; Dubberke, Erik R

    2015-03-01

    Clostridium difficile infection (CDI) is the most common cause of infectious health care-associated diarrhea and is a major burden to patients and the health care system. The incidence and severity of CDI remain at historically high levels. This article reviews the morbidity, mortality, and costs associated with CDI. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. 112 current trend in antimicrobial susceptibility pattern of clostridium ...

    African Journals Online (AJOL)

    DR. AMINU

    inorganic substances. The odour usually gives a suggestion that Clostridium species was present. C. tetani is an obligate anaerobe, and therefore its isolation requires an environment, which is totally devoid of even a trace of oxygen. The technique applied in this research to create an anaerobic condition was found to be ...

  11. First described case of prosthetic joint infection with Clostridium disporicum.

    Science.gov (United States)

    McBride, Joseph A; Sterkel, Alana K; Rehrauer, William M; Smith, Jeannina A

    2017-12-01

    An orthopedic hardware infection with Clostridium disporicum is described. C. disporicum is a gram positive anaerobic bacillus which can contain two subterminal spores. C. disporicum had not previously been reported in musculoskeletal infections. Gram stains demonstrating gram positive bacilli with two subterminal spores should alert practitioners to the possibility of C. disporicum infection. Published by Elsevier Ltd.

  12. 9 CFR 113.455 - Clostridium Perfringens Type D Antitoxin.

    Science.gov (United States)

    2010-01-01

    ... Clostridium perfringens Type D. Each serial shall be tested as provided in this section. Any serial found... following words and terms shall mean: (i) International antitoxin unit. (I.U.) That quantity of Epsilon... 0.25 gram of sodium chloride in each 100 ml of distilled water; adjusting the pH to 7.2; autoclaving...

  13. 9 CFR 113.454 - Clostridium Perfringens Type C Antitoxin.

    Science.gov (United States)

    2010-01-01

    ... Clostridium perfringens Type C. Each serial shall be tested as provided in this section. Any serial found... following words and terms shall mean: (i) International antitoxin unit. (I.U.) That quantity of Beta... chloride in each 100 ml of distilled water; adjusting the pH to 7.2; autoclaving at 250 °F. for 25 minutes...

  14. Update of treatment algorithms for Clostridium difficile infection

    NARCIS (Netherlands)

    Ooijevaar, R. E.; van Beurden, Y. H.; Terveer, E. M.; Goorhuis, A.; Bauer, M. P.; Keller, J. J.; Mulder, C. J. J.; Kuijper, E. J.

    2018-01-01

    Clostridium difficile is the leading cause of antibiotic-associated diarrhea, both in healthcare facilities and the community. The recurrence rate of C. difficile infection (CDI) remains high, up to 20%. Since the publication of the ESCMID guidance document on CDI treatment in 2014, new therapeutic

  15. FT-IR spectroscopic analysis for studying Clostridium cell response to conversion of enzymatically hydrolyzed hay

    Science.gov (United States)

    Grube, Mara; Gavare, Marita; Nescerecka, Alina; Tihomirova, Kristina; Mezule, Linda; Juhna, Talis

    2013-07-01

    Grass hay is one of assailable cellulose containing non-food agricultural wastes that can be used as a carbohydrate source by microorganisms producing biofuels. In this study three Clostridium strains Clostridium acetobutylicum, Clostridium beijerinckii and Clostridium tetanomorphum, capable of producing acetone, butanol and ethanol (ABE) were adapted to convert enzymatically hydrolyzed hay used as a growth media additive. The results of growth curves, substrate degradation kinetics and FT-IR analyses of bacterial biomass macromolecular composition showed diverse strain-specific cell response to the growth medium composition.

  16. Risk factors for Clostridium difficile infection in HIV-infected patients.

    Science.gov (United States)

    Imlay, Hannah; Kaul, Daniel; Rao, Krishna

    2016-01-01

    Clostridium difficile infection is a healthcare-associated infection resulting in significant morbidity. Although immunosuppression is associated with Clostridium difficile infection acquisition and adverse outcomes, the epidemiology of Clostridium difficile infection in HIV-infected patients has been little studied in the era of antiretroviral therapy. This study identifies the risk factors for acquisition of Clostridium difficile infection in HIV-infected patients. A retrospective, propensity score-matched case-control study design was employed, with patients selected from our institution's outpatient HIV clinic. Clostridium difficile infection cases were defined as having positive stool testing plus an appropriate clinical presentation. The propensity score was generated via multiple logistic regression from year of HIV diagnosis, age at first contact, duration of follow-up, gender, and initial CD4 count. The 46 cases included were matched to a total of 180 controls. Prior antibiotic treatment was a significant predictor of Clostridium difficile infection (odds ratio: 13, 95% confidence interval: 3.49-48.8, p  Clostridium difficile infection in the multivariable model (odds ratio: 15.17, confidence interval: 1.31-175.9, p  = .021). As in the general population, frequent hospitalizations and exposure to antimicrobials are independent predictors of Clostridium difficile infection acquisition in patients with HIV. Additionally, low CD4 count and proton pump inhibitor use are new potentially modifiable variables that can be targeted for prevention of Clostridium difficile infection in future interventional studies.

  17. Beneficial and harmful roles of bacteria from the Clostridium genus.

    Science.gov (United States)

    Samul, Dorota; Worsztynowicz, Paulina; Leja, Katarzyna; Grajek, Włodzimierz

    2013-01-01

    Bacteria of the Clostridium genus are often described only as a biological threat and a foe of mankind. However, many of them have positive properties and thanks to them they may be used in many industry branches (e.g., in solvents and alcohol production, in medicine, and also in esthetic cosmetology). During the last 10 years interest in application of C. botulinum and C. tetani in medicine significantly increased. Currently, the structure and biochemical properties of neurotoxins produced by these bacterial species, as well as possibilities of application of such toxins as botulinum as a therapeutic factor in humans, are being intensely researched. The main aim of this article is to demonstrate that bacteria from Clostridium spp. are not only pathogens and the enemy of humanity but they also have many important beneficial properties which make them usable among many chemical, medical, and cosmetic applications.

  18. Models for the study of Clostridium difficile infection

    Science.gov (United States)

    Best, Emma L.; Freeman, Jane; Wilcox, Mark H.

    2012-01-01

    Models of Clostridium difficile infection (C. difficile) have been used extensively for Clostridium difficile (C. difficile) research. The hamster model of C. difficile infection has been most extensively employed for the study of C. difficile and this has been used in many different areas of research, including the induction of C. difficile, the testing of new treatments, population dynamics and characterization of virulence. Investigations using in vitro models for C. difficile introduced the concept of colonization resistance, evaluated the role of antibiotics in C. difficile development, explored population dynamics and have been useful in the evaluation of C. difficile treatments. Experiments using models have major advantages over clinical studies and have been indispensible in furthering C. difficile research. It is important for future study programs to carefully consider the approach to use and therefore be better placed to inform the design and interpretation of clinical studies. PMID:22555466

  19. Calcium Montmorillonite-based dietary supplement attenuates Necrotic Enteritis induced by Eimeria maxima and Clostridium perfringens in broilers

    Science.gov (United States)

    We provide the first description of Dietary Supplement of sorbent minerals attenuates Necrotic Enteritis Induced by Eimeria maxima and Clostridium perfringens in Broilers. Necrotic enteritis (NE) is a poultry disease caused by Clostridium perfringens and characterized by severe intestinal necrosis....

  20. Fidaxomicin for the treatment of Clostridium difficile infections.

    Science.gov (United States)

    Whitman, Craig B; Czosnowski, Quinn A

    2012-02-01

    To evaluate the pharmacology, microbiology, safety, and efficacy of fidaxomicin for treatment of Clostridium difficile infections (CDI). Literature was identified through Ovid MEDLINE (1948-December 2011) and International Pharmaceutical Abstracts (1970-December 2011) using the search terms fidaxomicin, OPT-80, PAR-101, OP-118, difimicin, tiacumicin, lipiarmycin, Clostridium difficile, Clostridium difficile infection, Clostridium difficile-associated diarrhea, and cost. Drug monographs were retrieved from manufacturers' Web pages, and the Red Book component of Micromedex was used for cost information. All pertinent Phase 1, 2, and 3 studies published in English were included. Fidaxomicin is a macrocyclic compound bactericidal against C. difficile and inhibits toxin and spore production. It has poor oral absorption with high fecal concentrations. Available Phase 2 and 3 data with fidaxomicin 200 mg orally every 12 hours demonstrate similar effectiveness in treating CDI compared to oral vancomycin. Fidaxomicin was shown to have less frequency of recurrent infections. Adverse effects are uncommon and occur at similar rates as with oral vancomycin. The most frequently reported adverse effects are gastrointestinal, hematologic, and electrolyte disorders. Available data are lacking in several areas, including the efficacy and safety of fidaxomicin compared to established regimens for mild-to-moderate, life-threatening, and recurrent CDIs. The cost of a 10-day course of fidaxomicin is significantly more than that of metronidazole and vancomycin for treatment of mild-to-moderate CDI. Fidaxomicin appears to be an effective and safe alternative to oral vancomycin for treatment of mild-to-moderate and severe CDI. Data on its use compared to guideline-recommended therapies for mild-to-moderate and life-threatening CDI are needed. Further data assessing the cost-effectiveness of fidaxomicin are needed. Currently, it cannot be recommended over vancomycin for treatment of CDI

  1. Clostridium difficile infections in patients with severe burns

    Science.gov (United States)

    2011-01-01

    placards indicating that hand hygiene should involve soap and water. Periodic hand hygiene compliance surveys have indicated relatively consistent...care unit: epidemiology, costs, and colonization pressure. Infect Control Hosp Epidemiol 2007;28:123–30. [6] Marcon AP, Gamba MA, Vianna LA. Nosocomial ...Clostridium difficile infections in patients with severe burns§ Scott J. Crabtree a, Janelle L. Robertson a,b, Kevin K. Chung c, Evan M. Renz b,c

  2. Clostridium difficile-associated diarrhoea in infants and children

    OpenAIRE

    Vuletić Biljana; Ristanović Elizabeta; Marković Slavica; Rašković Zorica; Radlović Vladimir; Igrutinović Zoran

    2017-01-01

    Clostridium difficile (CD) is the most common cause of nosocomial diarrhea in adults with high rates of morbidity and mortality. The epidemiology of CD infection (CDI) has changed in the last few decades associated with increasing severity of the infection rate related to the occurrence of NAP1 hypervirulent strain and the emergence of the disease among ambulatory patients and the wider community. Although little is known about CDI in pediatric patients, CD is surprisingly recognized as an im...

  3. Economic evaluation of interventions designed to reduce Clostridium difficile infection.

    Science.gov (United States)

    Brain, David; Yakob, Laith; Barnett, Adrian; Riley, Thomas; Clements, Archie; Halton, Kate; Graves, Nicholas

    2018-01-01

    Healthcare decision-makers are increasingly expected to balance increasing demand for health services with a finite budget. The role of economic evaluation in healthcare is increasing and this research provides decision-makers with new information about the management of Clostridium difficile infection, from an economic perspective. A model-based economic evaluation was undertaken to identify the most cost-effective healthcare intervention relating to the reduction of Clostridium difficile transmission. Efficacy evidence was synthesised from the literature and was used to inform the effectiveness of both bundled approaches and stand-alone interventions, where appropriate intervention combinations were coupled together. Changes in health outcomes were estimated by combining information about intervention effectiveness and its subsequent impact on quality of life. A bundled approach of improving hand hygiene and environmental cleaning produces the best combination of increased health benefits and cost-savings. It has the highest mean net monetary benefit when compared to all other interventions. This intervention remains the optimal decision under different clinical circumstances, such as when mortality rate and patient length of stay are increased. Bundled interventions offered the best opportunity for health improvements. These findings provide healthcare decision-makers with novel information about the allocation of scarce resources relating to Clostridium difficile. If investments are not made in interventions that clearly yield gains in health outcomes, the allocation and use of scarce healthcare resources is inappropriate and improvements in health outcomes will be forgone.

  4. Economic evaluation of interventions designed to reduce Clostridium difficile infection.

    Directory of Open Access Journals (Sweden)

    David Brain

    Full Text Available Healthcare decision-makers are increasingly expected to balance increasing demand for health services with a finite budget. The role of economic evaluation in healthcare is increasing and this research provides decision-makers with new information about the management of Clostridium difficile infection, from an economic perspective.A model-based economic evaluation was undertaken to identify the most cost-effective healthcare intervention relating to the reduction of Clostridium difficile transmission. Efficacy evidence was synthesised from the literature and was used to inform the effectiveness of both bundled approaches and stand-alone interventions, where appropriate intervention combinations were coupled together. Changes in health outcomes were estimated by combining information about intervention effectiveness and its subsequent impact on quality of life.A bundled approach of improving hand hygiene and environmental cleaning produces the best combination of increased health benefits and cost-savings. It has the highest mean net monetary benefit when compared to all other interventions. This intervention remains the optimal decision under different clinical circumstances, such as when mortality rate and patient length of stay are increased. Bundled interventions offered the best opportunity for health improvements.These findings provide healthcare decision-makers with novel information about the allocation of scarce resources relating to Clostridium difficile. If investments are not made in interventions that clearly yield gains in health outcomes, the allocation and use of scarce healthcare resources is inappropriate and improvements in health outcomes will be forgone.

  5. SEVERE CLOSTRIDIUM DIFFICILE INFECTIONS. A SYSTEMATIC LITERATURE -review-

    Directory of Open Access Journals (Sweden)

    Adriana Elena NICA

    2016-06-01

    Full Text Available Clostridium difficile is a bacterium that has been brought to the attention of the medical community recently, as the number of infections related to it has increased dramatically. This is happening mainly because of the excessive and defective use of antibiotic therapy. The pathology of a Clostridium Difficile infection is very complex, as it ranges from easy symptoms like abdominal pain and diarrhea to severe complications, like toxic megacolon. The management of these infections has become even more difficult, as they are not appearing only in the hospital environment anymore, but also outside of it. The bacterium spreads through poor hands hygiene. Also, we don’t have a clear strategy for overcoming an infection like this, so it gets even more difficult as most of the times the doctors need to rely only on their experience and knowledge to find ways of battling it. We would like to underline the research opportunities that are available in this domain as very few things are known about Clostridium difficile and also the crucial importance of research, as these infections are common and dangerous not only for patients, but for the medical staff and their families too.

  6. Emerging therapies for Clostridium difficile infection – focus on fidaxomicin

    Directory of Open Access Journals (Sweden)

    Chaparro-Rojas F

    2013-06-01

    Full Text Available Fredy Chaparro-Rojas, Kathleen M MullaneDepartment of Medicine, Section of Infectious Diseases, University of Chicago, Chicago, IL, USAAbstract: The epidemiology of Clostridium difficile infections (CDI has evolved during the last decades, with an increase in the reported incidence, severity of cases, and rate of mortality and relapses. These increases have primarily affected some special populations including the elderly, patients requiring concomitant antibiotic therapy, patients with renal failure, and patients with cancer. Until recently, the treatment of CDI was limited to either metronidazole or vancomycin. New therapeutic options have emerged to address the shortcomings of current antibiotic therapy. Fidaxomicin stands out as the first-in-class oral macrocyclic antibiotic with targeted activity against C. difficile and minimal collateral damage on the normal colonic flora. Fidaxomicin has demonstrated performance not inferior to what is considered the “gold standard” available therapy for CDI, vancomycin, in two separate Phase III clinical trials, but with significant advantages, including fewer recurrences and higher rates of sustained clinical cures. Fidaxomicin constitutes an important development in targeted antibiotic therapy for CDI and must be considered as a first-line agent for patients with risk factors known to portend relapse and severe infection.Keywords: fidaxomicin, Clostridium difficile-associated diarrhea, CDAD, Clostridium difficile infection (CDI, vancomycin, metronidazole

  7. Detection of enterotoxigenic Clostridium perfringens in meat samples by using molecular methods.

    Science.gov (United States)

    Kaneko, Ikuko; Miyamoto, Kazuaki; Mimura, Kanako; Yumine, Natsuko; Utsunomiya, Hirotoshi; Akimoto, Shigeru; McClane, Bruce A

    2011-11-01

    To prevent food-borne bacterial diseases and to trace bacterial contamination events to foods, microbial source tracking (MST) methods provide important epidemiological information. To apply molecular methods to MST, it is necessary not only to amplify bacterial cells to detection limit levels but also to prepare DNA with reduced inhibitory compounds and contamination. Isolates carrying the Clostridium perfringens enterotoxin gene (cpe) on the chromosome or a plasmid rank among the most important food-borne pathogens. Previous surveys indicated that cpe-positive C. perfringens isolates are present in only ∼5% of nonoutbreak food samples and then only at low numbers, usually less than 3 cells/g. In this study, four molecular assays for the detection of cpe-positive C. perfringens isolates, i.e., ordinary PCR, nested PCR, real-time PCR, and loop-mediated isothermal amplification (LAMP), were developed and evaluated for their reliability using purified DNA. For use in the artificial contamination of meat samples, DNA templates were prepared by three different commercial DNA preparation kits. The four molecular assays always detected cpe when >10³ cells/g of cpe-positive C. perfringens were present, using any kit. Of three tested commercial DNA preparation kits, the InstaGene matrix kit appeared to be most suitable for the testing of a large number of samples. By using the InstaGene matrix kit, the four molecular assays efficiently detected cpe using DNA prepared from enrichment culture specimens of meat samples contaminated with low numbers of cpe-positive C. perfringens vegetative cells or spores. Overall, the current study developed molecular assay protocols for MST to detect the contamination of foods with low numbers of cells, and at a low frequency, of cpe-positive C. perfringens isolates.

  8. Detection of Enterotoxigenic Clostridium perfringens in Meat Samples by Using Molecular Methods▿

    Science.gov (United States)

    Kaneko, Ikuko; Miyamoto, Kazuaki; Mimura, Kanako; Yumine, Natsuko; Utsunomiya, Hirotoshi; Akimoto, Shigeru; McClane, Bruce A.

    2011-01-01

    To prevent food-borne bacterial diseases and to trace bacterial contamination events to foods, microbial source tracking (MST) methods provide important epidemiological information. To apply molecular methods to MST, it is necessary not only to amplify bacterial cells to detection limit levels but also to prepare DNA with reduced inhibitory compounds and contamination. Isolates carrying the Clostridium perfringens enterotoxin gene (cpe) on the chromosome or a plasmid rank among the most important food-borne pathogens. Previous surveys indicated that cpe-positive C. perfringens isolates are present in only ∼5% of nonoutbreak food samples and then only at low numbers, usually less than 3 cells/g. In this study, four molecular assays for the detection of cpe-positive C. perfringens isolates, i.e., ordinary PCR, nested PCR, real-time PCR, and loop-mediated isothermal amplification (LAMP), were developed and evaluated for their reliability using purified DNA. For use in the artificial contamination of meat samples, DNA templates were prepared by three different commercial DNA preparation kits. The four molecular assays always detected cpe when >103 cells/g of cpe-positive C. perfringens were present, using any kit. Of three tested commercial DNA preparation kits, the InstaGene matrix kit appeared to be most suitable for the testing of a large number of samples. By using the InstaGene matrix kit, the four molecular assays efficiently detected cpe using DNA prepared from enrichment culture specimens of meat samples contaminated with low numbers of cpe-positive C. perfringens vegetative cells or spores. Overall, the current study developed molecular assay protocols for MST to detect the contamination of foods with low numbers of cells, and at a low frequency, of cpe-positive C. perfringens isolates. PMID:21890671

  9. Examination of Clostridium difficile Contamination in beef meat distributed in Isfahan using culture and Multiplex-PCR method

    OpenAIRE

    zahra Esfandiari; Mohammad Jalali; Hamid Ezzatpanah; Scott Weese; Mohammad Chamani

    2014-01-01

    Introduction: With regard to increasing of community associated Clostridium difficile infection in recent years, the probable transmission of Clostridium difficile from food to human was supposed. Most of reports on this issue were allocated to examine the prevalence of Clostridium difficile in red meat. The current study aimed at examination of the prevalence of Clostridium difficile in beef meat. Materials and methods: A total of 100 beef meat samples includi...

  10. Evaluation of Correlation between Pretest Probability for Clostridium difficile Infection and Clostridium difficile Enzyme Immunoassay Results.

    Science.gov (United States)

    Kwon, Jennie H; Reske, Kimberly A; Hink, Tiffany; Burnham, C A; Dubberke, Erik R

    2017-02-01

    The objective of this study was to evaluate the clinical characteristics and outcomes of hospitalized patients tested for Clostridium difficile and determine the correlation between pretest probability for C. difficile infection (CDI) and assay results. Patients with testing ordered for C. difficile were enrolled and assigned a high, medium, or low pretest probability of CDI based on clinical evaluation, laboratory, and imaging results. Stool was tested for C. difficile by toxin enzyme immunoassay (EIA) and toxigenic culture (TC). Chi-square analyses and the log rank test were utilized. Among the 111 patients enrolled, stool samples from nine were TC positive and four were EIA positive. Sixty-one (55%) patients had clinically significant diarrhea, 19 (17%) patients did not, and clinically significant diarrhea could not be determined for 31 (28%) patients. Seventy-two (65%) patients were assessed as having a low pretest probability of having CDI, 34 (31%) as having a medium probability, and 5 (5%) as having a high probability. None of the patients with low pretest probabilities had a positive EIA, but four were TC positive. None of the seven patients with a positive TC but a negative index EIA developed CDI within 30 days after the index test or died within 90 days after the index toxin EIA date. Pretest probability for CDI should be considered prior to ordering C. difficile testing and must be taken into account when interpreting test results. CDI is a clinical diagnosis supported by laboratory data, and the detection of toxigenic C. difficile in stool does not necessarily confirm the diagnosis of CDI. Copyright © 2017 American Society for Microbiology.

  11. Sporulation of Clostridium cylindrosporum on a Defined, Low-Manganese Medium

    OpenAIRE

    Sacks, L. E.; Smith, M. R.

    1987-01-01

    Clostridium cylindrosporum HC-1 grew and sporulated well on a defined medium. This is the first demonstration of sporulation of a purinolytic clostridium on a defined medium; manganese levels were below those considered essential for sporulation of most Bacillus species. Sporulation appeared to be initiated before exhaustion of the purine substrate.

  12. Mathematical modeling and growth kinetics of Clostridium sporogenes in cooked beef

    Science.gov (United States)

    Clostridium sporogenes PA 3679 is a common surrogate for proteolytic Clostridium botulinum for thermal process development and validation. However, little information is available concerning the growth kinetics of C. sporogenes in food. Therefore, the objective of this study was to investigate the...

  13. Draft Genome Sequence of Antimicrobial-Producing Clostridium sp. JC272, Isolated from Marine Sediment

    OpenAIRE

    Tushar, L.; Sasi Jyothsna, T. S.; Sasikala, C.; Ramana, C. V.

    2015-01-01

    We announce the draft genome sequence of Clostridium sp. JC272, isolated from a sediment sample collected from marine habitats of Gujarat, India. Clostridium sp. JC272 is an obligate anaerobe and has the ability to produce antimicrobial compounds. The genome sequence indicates the strain?s capability of producing small peptides (microcins), which are potential novel antibiotics.

  14. Hydrolytic bacteria in mesophilic and thermophilic degradation of plant biomass

    Energy Technology Data Exchange (ETDEWEB)

    Zverlov, Vladimir V.; Hiegl, Wolfgang; Koeck, Daniela E.; Koellmeier, Tanja; Schwarz, Wolfgang H. [Department of Microbiology, Technische Universitaet Muenchen, Freising-Weihenstephan (Germany); Kellermann, Josef [Max Planck Institute for Biochemistry, Am Klopferspitz, Martinsried (Germany)

    2010-12-15

    Adding plant biomass to a biogas reactor, hydrolysis is the first reaction step in the chain of biological events towards methane production. Maize silage was used to enrich efficient hydrolytic bacterial consortia from natural environments under conditions imitating those in a biogas plant. At 55-60 C a more efficient hydrolyzing culture could be isolated than at 37 C. The composition of the optimal thermophilic bacterial consortium was revealed by sequencing clones from a 16S rRNA gene library. A modified PCR-RFLP pre-screening method was used to group the clones. Pure anaerobic cultures were isolated. 70% of the isolates were related to Clostridium thermocellum. A new culture-independent method for identification of cellulolytic enzymes was developed using the isolation of cellulose-binding proteins. MALDI-TOF/TOF analysis and end-sequencing of peptides from prominent protein bands revealed cellulases from the cellulosome of C. thermocellum and from a major cellulase of Clostridium stercorarium. A combined culture of C. thermocellum and C. stercorarium was shown to excellently degrade maize silage. A spore preparation method suitable for inoculation of maize silage and optimal hydrolysis was developed for the thermophilic bacterial consortium. This method allows for concentration and long-term storage of the mixed culture for instance for inoculation of biogas fermenters. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  15. Clostridium septicum aortitis with synchronous ascending colon and rectal adenocarcinoma.

    Science.gov (United States)

    Jain, Deepanshu; Kistler, Andrew C; Kozuch, Patricia

    2017-01-01

    Clostridium septicum ( C. septicum ) aortitis is a rare condition frequently associated with colon adenocarcinoma and carries a poor prognosis. We report the case of a 66-year-old man who presented with abdominal pain, blood in the stool, fever and chills. Laboratory tests were significant for leukocytosis and microcytic anemia. Abdominal imaging revealed a right colon mass and aortitis. Colonoscopy confirmed the right colon mass and also discovered a rectal mass, both adenocarcinomas. Treatment consisted of antibiotics, aortic repair, right hemi-colectomy and later trans-anal excision of the rectal mass. Blood cultures and the aortic specimen grew C. septicum . The patient improved and was doing well in follow up.

  16. Probiotics for the treatment of Clostridium difficile associated disease

    OpenAIRE

    Fitzpatrick, Leo R

    2013-01-01

    The purpose of this review paper is to update the current and potential future role of probiotics for Clostridium difficile-associated disease (CDAD). Included in this review, is an update on the testing of newer probiotics (e.g., Bacillus coagulans GBI-30, 6086) in animal models of CDAD. There is a focus on the modulation of signal transduction pathways (i.e., transcription factors like cAMP response element-binding, activator protein 1, and nuclear factor kappa B), as well as the inhibition...

  17. Probiotics and Antibiotic-Associated Diarrhea and Clostridium difficile Infection

    Science.gov (United States)

    Surawicz, Christina M.

    Diarrhea is a common side effect of antibiotics. Antibiotics can cause diarrhea in 5-25% of individuals who take them but its occurrence is unpredictable. Diarrhea due to antibiotics is called antibiotic-associated diarrhea (AAD). Diarrhea may be mild and resolve when antibiotics are discontinued, or it may be more severe. The most severe form of AAD is caused by overgrowth of Clostridium difficile which can cause severe diarrhea, colitis, pseudomembranous colitis, or even fatal toxic megacolon. Rates of diarrhea vary with the specific antibiotic as well as with the individual susceptibility.

  18. Chitinolytic enzymes from Clostridium paraputrificum for biomedical applications

    Czech Academy of Sciences Publication Activity Database

    Kolenko, Petr; Dušková, Jarmila; Tiščenko, Galina; Koval, Tomáš; Fejfarová, Karla; Šimůnek, Jiří; Hašek, Jindřich; Dohnálek, Jan

    2014-01-01

    Roč. 281, Supplement s1 (2014), s. 653 ISSN 1742-464X. [FEBS EMBO 2014 Conference. 30.08.2014-04.09.2014, Paris] R&D Projects: GA MŠk(CZ) EE2.3.30.0029; GA MŠk LG14009; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61389013 ; RVO:86652036 ; RVO:67985904 Keywords : chitin * chitinase * clostridium Subject RIV: CE - Biochemistry; EB - Genetics ; Molecular Biology (BTO-N); EE - Microbiology, Virology (UZFG-Y) http://onlinelibrary.wiley.com/doi/10.1111/febs.12919/abstract

  19. Pseudomembranous Colitis: Not Always Caused by Clostridium difficile

    Directory of Open Access Journals (Sweden)

    Derek M. Tang

    2014-01-01

    Full Text Available Although classically pseudomembranous colitis is caused by Clostridium difficile, it can result from several etiologies. Certain medications, chemical injury, collagenous colitis, inflammatory bowel disease, ischemia, and other infectious pathogens can reportedly cause mucosal injury and subsequent pseudomembrane formation. We present the case of a middle-aged woman with vascular disease who was incorrectly diagnosed with refractory C. difficile infection due to the presence of pseudomembranes. Further imaging, endoscopy, and careful histopathology review revealed chronic ischemia as the cause of her pseudomembranous colitis and diarrhea. This case highlights the need for gastroenterologists to consider non-C. difficile etiologies when diagnosing pseudomembranous colitis.

  20. Clostridium difficile-ribotype 027 er en udfordring

    DEFF Research Database (Denmark)

    Nyboe Sommer, Trine; Ravn, Pernille; Gjørup, Ida

    2014-01-01

    Infection with Clostridium difficile is the primary infective cause of antibiotic-associated diarrhoea. In 2008, a major outbreak of CD027 took place in North Zealand, Denmark. We described this infection in a single medical department. Patients positive for C. difficile enlisted at Medical...... Department O, Herlev Hospital, in 2009 were included and demographic data were recorded. In total, 69 patients were included, average age 83 years, Charlson Comorbidity Score 4. Of all patients 24 died. Further studies are needed in order to treat and minimize infection with C. difficile....

  1. Controversies Surrounding Clostridium difficile Infection in Infants and Young Children

    Directory of Open Access Journals (Sweden)

    Maribeth R. Nicholson

    2014-06-01

    Full Text Available Clostridium difficile is a frequent cause of antibiotic-associated diarrhea in adults and older children. However, as many as 80% of infants can be asymptomatically colonized. The reasons for this have not been well established but are believed to be due to differences in toxin receptors or toxin internalization. Determining which children who test positive for C. difficile warrant treatment is exceedingly difficult, especially in the setting of increased rates of detection and the rising risk of disease in children lacking classic risk factors for C. difficile.

  2. Molecular diversity of neurotoxins from Clostridium botulinum type D strains.

    OpenAIRE

    Moriishi, K; Syuto, B; Kubo, S; Oguma, K

    1989-01-01

    The molecular properties of Clostridium botulinum type D South African (D-SA) were compared with those of neurotoxins from type D strain 1873 (D-1873) and type C strains Stockholm and 6813. D-SA toxin, purified 610-fold from the culture supernatant in an overall yield of 30%, consisted of an intact peptide chain with a molecular weight of 140,000. Limited proteolysis of the toxin by trypsin formed a dichain structure consisting of a light chain (Mr, 50,000) and a heavy chain (Mr, 90,000) link...

  3. Conserved oligopeptide permeases modulate sporulation initiation in Clostridium difficile.

    Science.gov (United States)

    Edwards, Adrianne N; Nawrocki, Kathryn L; McBride, Shonna M

    2014-10-01

    The anaerobic gastrointestinal pathogen Clostridium difficile must form a metabolically dormant spore to survive in oxygenic environments and be transmitted from host to host. The regulatory factors by which C. difficile initiates and controls the early stages of sporulation in C. difficile are not highly conserved in other Clostridium or Bacillus species. Here, we investigated the role of two conserved oligopeptide permeases, Opp and App, in the regulation of sporulation in C. difficile. These permeases are known to positively affect sporulation in Bacillus species through the import of sporulation-specific quorum-sensing peptides. In contrast to other spore-forming bacteria, we discovered that inactivating these permeases in C. difficile resulted in the earlier expression of early sporulation genes and increased sporulation in vitro. Furthermore, disruption of opp and app resulted in greater virulence and increased the amounts of spores recovered from feces in the hamster model of C. difficile infection. Our data suggest that Opp and App indirectly inhibit sporulation, likely through the activities of the transcriptional regulator SinR and its inhibitor, SinI. Taken together, these results indicate that the Opp and App transporters serve a different function in controlling sporulation and virulence in C. difficile than in Bacillus subtilis and suggest that nutrient availability plays a significant role in pathogenesis and sporulation in vivo. This study suggests a link between the nutritional status of the environment and sporulation initiation in C. difficile. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  4. Role of collagenase clostridium histolyticum in Peyronie's disease

    Directory of Open Access Journals (Sweden)

    Peak TC

    2015-09-01

    Full Text Available Taylor C Peak,1 Gregory C Mitchell,2 Faysal A Yafi,2 Wayne J Hellstrom2 1Department of Urology, Tulane University School of Medicine, 2Section of Andrology, Department of Urology, Tulane University School of Medicine, New Orleans, LA, USA Abstract: Peyronie's disease is a localized connective tissue disease characterized by an active, inflammatory phase and a stable, quiescent phase, with the eventual development of collagenous plaques within the tunica albuginea of the penis. Risk factors primarily associated with Peyronie's disease include Dupuytren's contracture, penile trauma, and family history. A variety of treatment strategies have been utilized, including oral and topical agents, electromotive drug administration, intralesional injections, extracorporeal shockwave therapy, penile traction, and surgery. However, most of these strategies are ineffective, with surgery being the only definitive treatment. Collagenase clostridium histolyticum is a newly US Food and Drug Administration-approved agent for intralesional injection. It is thought to downregulate many of the disease-related genes, cytokines, and growth factors and degrade collagen fibers. It also suppresses cell attachment, spreading, and proliferation. Collagenase clostridium histolyticum has been clinically proven to be a safe and effective therapeutic option, demonstrating decreases in penile curvature and plaque consistency, as well as increases in patient satisfaction. During clinical evaluation, the Peyronie's Disease Questionnaire was validated as an effective tool for assessing treatment outcomes. Keywords: connective tissue disease, CCH, Xiaflex, Peyronie's Disease Questionnaire

  5. CRYSTAL STRUCTURE OF CLOSTRIDIUM BOTULINUM NEUROTOXIN SEROTYPE B

    International Nuclear Information System (INIS)

    SWAMINATHAN, S.; ESWARAMOORTHY, S.

    2001-01-01

    The toxigenic strains of Clostridium botulinum produce seven serologically distinct types of neurotoxins labeled A - G (EC 3.4.24.69), while Clostridium tetani produces tetanus neurotoxin (EC 3.4.24.68). Botulinum and tetanus neurotoxins (BoNTs and TeNT) are produced as single inactive chains of molecular mass of approximately 150 kDa. Most of these neurotoxins are released after being cleaved into two chains, a heavy chain (HI) of 100 kDa and a light chain (L) of 50 kDa held together by an interchain disulfide bond, by tissue proteinases. BoNT/E is released as a single chain but cleaved by host proteinases[1]. Clostvidium botulinum neurotoxins are extremely poisonous proteins with their LD(sub 50) for humans in the range of 0.1 - 1 ng kg(sup -1)[2]. Botulinum neurotoxins are responsible for neuroparalytic syndromes of botulism characterized by serious neurological disorders and flaccid paralysis. BoNTs block the release of acetylcholine at the neuromuscular junction causing flaccid paralysis while TeNT blocks the release of neurotransmitters like glycine and(gamma)-aminobutyric acid (GABA) in the inhibitory interneurons of the spinal cord resulting in spastic paralysis. In spite of different clinical symptoms, their aetiological agents intoxicate neuronal cells in the same way and these toxins have similar structural organization[3

  6. Management of Clostridium difficile in a developing nation

    Directory of Open Access Journals (Sweden)

    Azadeh Nasrollah

    2016-01-01

    Full Text Available Introduction: Clostridium difficile is the most important definable cause of healthcare acquired diarrhea. Recommended treatments for Clostridium difficile infection (CDI are metronidazole, oral vancomycin and fidaxomicin (a new narrow spectrum macrocyclic antibiotic. Aim: The aim of this investigation was to review the treatment of CDI in Iran. Method: 1600 medical records and prescriptions were scrutinized for patients complaining of diarrhea, colitis and gastroenteritis. The therapeutic route was investigated in each individual case bearing in mind the medical and medication history as well as other co-morbidities. Results: The selection of antibiotic by many medical practitioners for the treatment of diarrhea, colitis and gastroenteritis were inappropriate and random. In most cases the chosen antibiotic, can itself be associated with initiation or worsening of CDI. Conclusion: The needs for antimicrobial stewardship program to preserve the effectiveness of current available therapies are strongly recommended. This program must focus on the overall reduction of inappropriate antibiotic prescribing and ultimately on enforcing the adherence to the reputable antibacterial guidelines.

  7. CRYSTAL STRUCTURE OF CLOSTRIDIUM BOTULINUM NEUROTOXIN SEROTYPE B.

    Energy Technology Data Exchange (ETDEWEB)

    SWAMINATHAN,S.; ESWARAMOORTHY,S.

    2001-11-19

    The toxigenic strains of Clostridium botulinum produce seven serologically distinct types of neurotoxins labeled A - G (EC 3.4.24.69), while Clostridium tetani produces tetanus neurotoxin (EC 3.4.24.68). Botulinum and tetanus neurotoxins (BoNTs and TeNT) are produced as single inactive chains of molecular mass of approximately 150 kDa. Most of these neurotoxins are released after being cleaved into two chains, a heavy chain (HI) of 100 kDa and a light chain (L) of 50 kDa held together by an interchain disulfide bond, by tissue proteinases. BoNT/E is released as a single chain but cleaved by host proteinases [1]. Clostvidium botulinum neurotoxins are extremely poisonous proteins with their LD{sub 50} for humans in the range of 0.1 - 1 ng kg{sup -1} [2]. Botulinum neurotoxins are responsible for neuroparalytic syndromes of botulism characterized by serious neurological disorders and flaccid paralysis. BoNTs block the release of acetylcholine at the neuromuscular junction causing flaccid paralysis while TeNT blocks the release of neurotransmitters like glycine and {gamma}-aminobutyric acid (GABA) in the inhibitory interneurons of the spinal cord resulting in spastic paralysis. In spite of different clinical symptoms, their aetiological agents intoxicate neuronal cells in the same way and these toxins have similar structural organization [3].

  8. Clostridium Difficile Infection Complicated By Toxic Megacolon In Immunocompetent Patient

    Directory of Open Access Journals (Sweden)

    Draganescu Miruna

    2017-02-01

    Full Text Available Toxic megacolon can be a form of severe clinical course of the infection with Clostridium difficile (ICD, life-threatening, requiring a particular course of treatment. Infection with Clostridium difficile in the Galati Infectious Disease Hospital presents rising number of cases, namely 172 cases in 2014, 271 cases in 2015 and 301 cases in 2016 with clinical evolutions with different severity degrees, including toxic megacolon and death. Among 744 patients with ICD in our clinic, since 1st January 2014 to 31 December 2016. The frequency of toxic megacolon (TM was 0,537%, so: 3 toxic megacolon cases with favorable evolution with treatment with vancomycin and metronidazole and just one case whose evolution was aggravated under this therapy and evolved favorably under treatment with tigecycline. The work presents this last case of ICD occurred in a 69 years old, immunocompetent man with unknown concomitant chronic diseases which undergoes surgery for bilateral inguinal hernia and receives antibiotherapy with cephalosporin IIIrd generation during surgery and after 7 days develops medium degree ICD with score Atlas 3 and receives therapy with oral vancomycin. He presents clinical aggravation during this therapy with the occurrence of colon dilatation, ascites and right pleurisy at ultrasound and therapy associated with metronidazole is decided. Clinical aggravation continues in this combined therapy with defining the clinical, colonoscopy and tomography criteria for TM and is decided surgical monitoring and replacing antibiotherapy with tigecycline. Evolution is favorable with tigecycline without surgical intervention.

  9. Clostridium difficile infection in solid organ transplant recipients.

    Science.gov (United States)

    Nanayakkara, Deepa; Nanda, Neha

    2017-08-01

    Clostridium difficile infection (CDI) is a major healthcare-associated infection that causes significant morbidity and an economic impact in the United States. In this review, we provide an overview of Clostridium difficile infection in solid organ transplant recipients with an emphasis on recent literature. C. difficile in solid organ transplant population has unique risk factors. Fecal microbiota transplantation has shown favorable results in treatment of recurrent C. difficile in this population. Preliminary data from animal studies suggests excellent efficacy with immunization against C. difficile toxins. Over the last decade, number of individuals receiving solid organ transplants has increased exponentially making peri-transplant complications a common occurrence.C. difficile is a frequent cause of morbidity in solid organ transplant recipients. Early and accurate diagnosis of C. difficile requires a stepwise approach. Differentiating between asymptomatic carriage and infection is a diagnostic challenge. Microbial diversity is inversely proportional to risk of C. difficile infection. Antimicrobial stewardship programs help to retain microbial diversity in individuals susceptible to CDI. Recurrent or relapsing C. difficile infection require fecal microbiota transplantation for definitive cure.

  10. Uterine Sarcoma Presenting with Sepsis from Clostridium perfringens Endometritis in a Postmenopausal Woman

    Directory of Open Access Journals (Sweden)

    Mary J. Kao

    2018-01-01

    Full Text Available Clostridium perfringens is an anaerobic gram positive rod that is found in normal vaginal and cervical flora in 1–10% of healthy women. Uterine infection with Clostridium perfringens is seen rarely but is often related to underlying uterine pathology and can progress quickly to sepsis. Early recognition of sepsis, prompt treatment with antibiotics, and source control with surgical management allow for optimal chance of recovery. We present a case of a postmenopausal woman who presented with sepsis, vaginal bleeding, and back pain who was found to have Clostridium perfringens infection in the setting of undifferentiated uterine sarcoma.

  11. Clostridium perfringens in London, July 2009: two weddings and an outbreak.

    Science.gov (United States)

    Eriksen, J; Zenner, D; Anderson, S R; Grant, K; Kumar, D

    2010-06-24

    Food poisoning outbreaks caused by Clostridium perfringens enterotoxin occur occasionally in Europe but have become less common in recent years. This paper presents the microbiological and epidemiological results of a large C. perfringens outbreak occurring simultaneously at two weddings that used the same caterer. The outbreak involved several London locations and required coordination across multiple agencies. A case-control study (n=134) was carried out to analyse possible associations between the food consumed and becoming ill. Food, environmental and stool samples were tested for common causative agents, including enterotoxigenic C. perfringens. The clinical presentation and the epidemiological findings were compatible with C. perfringens food poisoning and C. perfringens enterotoxin was detected in stool samples from two cases. The case-control study found statistically significant associations between becoming ill and eating either a specific chicken or lamb dish prepared by the same food handler of the implicated catering company. A rapid outbreak investigation with preliminary real-time results and the successful collaboration between the agencies and the caterer led to timely identification and rectification of the failures in the food handling practices.

  12. Bayesian modeling of Clostridium perfringens growth in beef-in-sauce products.

    Science.gov (United States)

    Jaloustre, S; Cornu, M; Morelli, E; Noël, V; Delignette-Muller, M L

    2011-04-01

    Models on Clostridium perfringens growth which have been published to date have all been deterministic. A probabilistic model describing growth under non-isothermal conditions was thus proposed for predicting C. perfringens growth in beef-in-sauce products cooked and distributed in a French hospital. Model parameters were estimated from different types of data from various studies. A Bayesian approach was proposed to model the overall uncertainty regarding parameters and potential variability on the 'work to be done' (h(0)) during the germination, outgrowth and lag phase. Three models which differed according to their description of this parameter h(0) were tested. The model with inter-curve variability on h(0) was found to be the best one, on the basis of goodness-of-fit assessment and validation with literature data on results obtained under non-isothermal conditions. This model was used in two-dimensional Monte Carlo simulations to predict C. perfringens growth throughout the preparation of beef-in-sauce products, using temperature profiles recorded in a hospital kitchen. The median predicted growth was 7.8×10(-2) log(10) cfu·g(-1) (95% credibility interval [2.4×10(-2), 0.8]) despite the fact that for more than 50% of the registered temperature profiles cooling steps were longer than those required by French regulations. Copyright © 2010 Elsevier Ltd. All rights reserved.

  13. In vitro reconstitution of the Clostridium botulinum type D progenitor toxin.

    Science.gov (United States)

    Kouguchi, Hirokazu; Watanabe, Toshihiro; Sagane, Yoshimasa; Sunagawa, Hiroyuki; Ohyama, Tohru

    2002-01-25

    Clostridium botulinum type D strain 4947 produces two different sizes of progenitor toxins (M and L) as intact forms without proteolytic processing. The M toxin is composed of neurotoxin (NT) and nontoxic-nonhemagglutinin (NTNHA), whereas the L toxin is composed of the M toxin and hemagglutinin (HA) subcomponents (HA-70, HA-17, and HA-33). The HA-70 subcomponent and the HA-33/17 complex were isolated from the L toxin to near homogeneity by chromatography in the presence of denaturing agents. We were able to demonstrate, for the first time, in vitro reconstitution of the L toxin formed by mixing purified M toxin, HA-70, and HA-33/17. The properties of reconstituted and native L toxins are indistinguishable with respect to their gel filtration profiles, native-PAGE profiles, hemagglutination activity, binding activity to erythrocytes, and oral toxicity to mice. M toxin, which contained nicked NTNHA prepared by treatment with trypsin, could no longer be reconstituted to the L toxin with HA subcomponents, whereas the L toxin treated with proteases was not degraded into M toxin and HA subcomponents. We conclude that the M toxin forms first by assembly of NT with NTNHA and is subsequently converted to the L toxin by assembly with HA-70 and HA-33/17.

  14. Hydrogen production from cellulose in a two-stage process combining fermentation and electrohydrogenesis

    KAUST Repository

    Lalaurette, Elodie; Thammannagowda, Shivegowda; Mohagheghi, Ali; Maness, Pin-Ching; Logan, Bruce E.

    2009-01-01

    A two-stage dark-fermentation and electrohydrogenesis process was used to convert the recalcitrant lignocellulosic materials into hydrogen gas at high yields and rates. Fermentation using Clostridium thermocellum produced 1.67 mol H2/mol

  15. Struggling with recurrent Clostridium difficile infections: is donor faeces the solution?

    NARCIS (Netherlands)

    van Nood, E.; Speelman, P.; Kuijper, E. J.; Keller, J. J.

    2009-01-01

    Patients with recurrent Clostridium difficile infections (CDI) in hospitals and the community constitute an increasing treatment problem. While most patients with a first infection respond to either metronidazole or oral vancomycin, therapy in recurrent C. difficile infections tends to fail

  16. Increase in Clostridium difficile-related Mortality Rates, United States, 1999-2004

    Centers for Disease Control (CDC) Podcasts

    Deaths related to Clostridium difficile are on the rise in the United States. Matthew Redelings from the Los Angeles County Department of Health discusses the increase and what can be done to prevent this infection.

  17. Draft Genome Sequence of Clostridium mangenotii TR, Isolated from the Fecal Material of a Timber Rattlesnake

    Science.gov (United States)

    Cochran, Philip A.; Dowd, Scot E.; Andersen, Kylie; Anderson, Nichole; Brennan, Rachel; Brook, Nicole; Callaway, Tracie; Diamante, Kimberly; Duberstine, Annie; Fitch, Karla; Freiheit, Heidi; Godlewski, Chantel; Gorman, Kelly; Haubrich, Mark; Hernandez, Mercedes; Hirtreiter, Amber; Ivanoski, Beth; Jaminet, Xochitl; Kirkpatrick, Travis; Kratowicz, Jennifer; Latus, Casey; Leable, Tiegen; Lingafelt, Nicole; Lowe, DeAnna; Lowrance, Holly; Malsack, Latiffa; Mazurkiewicz, Julie; Merlos, Persida; Messley, Jamie; Montemurro, Dawn; Nakitare, Samora; Nelson, Christine; Nye, Amber; Pazera, Valerie; Pierangeli, Gina; Rellora, Ashley; Reyes, Angelica; Roberts, Jennifer; Robins, Shadara; Robinson, Jeshannah; Schultz, Alissa; Seifert, Sara; Sigler, Elona; Spangler, Julie; Swift, Ebony; TenCate, Rebecca; Thurber, Jessica; Vallee, Kristin; Wamboldt, Jennifer; Whitten, Shannon; Woods, De’andrea; Wright, Amanda; Yankunas, Darin

    2014-01-01

    Here, we report the draft genome sequence of Clostridium mangenotii strain TR, which was isolated from the fecal material of a timber rattlesnake. This bacterium is nonpathogenic but contains 68 genes involved in virulence, disease, and defense. PMID:24407632

  18. Impact of end stage kidney disease on costs and outcomes of Clostridium difficile infection

    Directory of Open Access Journals (Sweden)

    Abhinav Goyal

    2017-09-01

    Conclusions: The presence of end stage kidney disease in hospitalized patients with Clostridium difficile infection is associated with higher mortality, a longer length of stay, and a higher cost of hospitalization.

  19. Reset of a critically disturbed microbial ecosystem: faecal transplant in recurrent Clostridium difficile infection

    NARCIS (Netherlands)

    Fuentes, Susana; van Nood, Els; Tims, Sebastian; Heikamp-de Jong, Ineke; ter Braak, Cajo J. F.; Keller, Josbert J.; Zoetendal, Erwin G.; de Vos, Willem M.

    2014-01-01

    Recurrent Clostridium difficile infection (CDI) can be effectively treated by infusion of a healthy donor faeces suspension. However, it is unclear what factors determine treatment efficacy. By using a phylogenetic microarray platform, we assessed composition, diversity and dynamics of faecal

  20. The Purine-Utilizing Bacterium Clostridium acidurici 9a: A Genome-Guided Metabolic Reconsideration

    OpenAIRE

    Hartwich, Katrin; Poehlein, Anja; Daniel, Rolf

    2012-01-01

    Clostridium acidurici is an anaerobic, homoacetogenic bacterium, which is able to use purines such as uric acid as sole carbon, nitrogen, and energy source. Together with the two other known purinolytic clostridia C. cylindrosporum and C. purinilyticum, C. acidurici serves as a model organism for investigation of purine fermentation. Here, we present the first complete sequence and analysis of a genome derived from a purinolytic Clostridium. The genome of C. acidurici 9a consists ...

  1. Multifunctional cellulase and hemicellulase

    Energy Technology Data Exchange (ETDEWEB)

    Fox, Brian G.; Takasuka, Taichi; Bianchetti, Christopher M.

    2015-09-29

    A multifunctional polypeptide capable of hydrolyzing cellulosic materials, xylan, and mannan is disclosed. The polypeptide includes the catalytic core (cc) of Clostridium thermocellum Cthe_0797 (CelE), the cellulose-specific carbohydrate-binding module CBM3 of the cellulosome anchoring protein cohesion region (CipA) of Clostridium thermocellum (CBM3a), and a linker region interposed between the catalytic core and the cellulose-specific carbohydrate binding module. Methods of using the multifunctional polypeptide are also disclosed.

  2. The potential economic value of screening hospital admissions for Clostridium difficile.

    Science.gov (United States)

    Bartsch, S M; Curry, S R; Harrison, L H; Lee, B Y

    2012-11-01

    Asymptomatic Clostridium difficile carriage has a prevalence reported as high as 51-85 %; with up to 84 % of incident hospital-acquired infections linked to carriers. Accurately identifying carriers may limit the spread of Clostridium difficile. Since new technology adoption depends heavily on its economic value, we developed an analytic simulation model to determine the cost-effectiveness screening hospital admissions for Clostridium difficile from the hospital and third party payer perspectives. Isolation precautions were applied to patients testing positive, preventing transmission. Sensitivity analyses varied Clostridium difficile colonization rate, infection probability among secondary cases, contact isolation compliance, and screening cost. Screening was cost-effective (i.e., incremental cost-effectiveness ratio [ICER] ≤ $50,000/QALY) for every scenario tested; all ICER values were ≤ $256/QALY. Screening was economically dominant (i.e., saved costs and provided health benefits) with a ≥10.3 % colonization rate and ≥5.88 % infection probability when contact isolation compliance was ≥25 % (hospital perspective). Under some conditions screening led to cost savings per case averted (range, $53-272). Clostridium difficile screening, coupled with isolation precautions, may be a cost-effective intervention to hospitals and third party payers, based on prevalence. Limiting Clostridium difficile transmission can reduce the number of infections, thereby reducing its economic burden to the healthcare system.

  3. Clostridium difficile in Food and Animals: A Comprehensive Review.

    Science.gov (United States)

    Rodriguez, C; Taminiau, B; Van Broeck, J; Delmée, M; Daube, G

    2016-01-01

    Zoonoses are infections or diseases that can be transmitted between animals and humans through direct contact, close proximity or the environment. Clostridium difficile is ubiquitous in the environment, and the bacterium is able to colonise the intestinal tract of both animals and humans. Since domestic and food animals frequently test positive for toxigenic C. difficile, even without showing any signs of disease, it seems plausible that C. difficile could be zoonotic. Therefore, animals could play an essential role as carriers of the bacterium. In addition, the presence of the spores in different meats, fish, fruits and vegetables suggests a risk of foodborne transmission. This review summarises the current available data on C. difficile in animals and foods, from when the bacterium was first described up to the present.

  4. ELIMINATION OF CLOSTRIDIUM PERFRINGENS DURING SURPLUS ACTIVATED SLUDGE HANDLING

    Directory of Open Access Journals (Sweden)

    Klaudiusz Grűbel

    2014-10-01

    Basis on the results of the research was concluded that microwave radiation (700W and 900W shows disintegration action expressed in COD value in the supernatant increase: 12 times increase value of COD with power 700W and 13 times for 900W radiation power. Electromagnetic wave contributed to partial higienisation of surplus activated sludge. The number of Clostridium perfringens decrease about 52% and 56% during the 120s of higienisation process with power 700W and 900W, respectively. Reduction of the overall number of bacteria under the influence of microwave radiation was 42% and 51% (respectively for 700W and 900W, and sticks from the family Enterobacteriaceae from 54% to 70% depending on the power of radiation, the time of operation and biochemical properties.

  5. Disparate subcellular location of putative sortase substrates in Clostridium difficile.

    Science.gov (United States)

    Peltier, Johann; Shaw, Helen A; Wren, Brendan W; Fairweather, Neil F

    2017-08-23

    Clostridium difficile is a gastrointestinal pathogen but how the bacterium colonises this niche is still little understood. Sortase enzymes covalently attach specific bacterial proteins to the peptidoglycan cell wall and are often involved in colonisation by pathogens. Here we show C. difficile proteins CD2537 and CD3392 are functional substrates of sortase SrtB. Through manipulation of the C-terminal regions of these proteins we show the SPKTG motif is essential for covalent attachment to the cell wall. Two additional putative substrates, CD0183 which contains an SPSTG motif, and CD2768 which contains an SPQTG motif, are not cleaved or anchored to the cell wall by sortase. Finally, using an in vivo asymmetric cleavage assay, we show that despite containing a conserved SPKTG motif, in the absence of SrtB these proteins are localised to disparate cellular compartments.

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

    Science.gov (United States)

    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.

  7. Infección por Clostridium tetani: sospecharla para diagnosticarla

    Directory of Open Access Journals (Sweden)

    Mónica Ríos Prego

    2016-12-01

    Full Text Available La infección por Clostridium tetani es una patología con un amplio periodo de incubación, que posee diversos modos de presentación clínica. Este hecho, junto con la baja incidencia de esta infección en los países industrializados , así como no poseer una técnica diagnóstica precisa que podamos obtener en un corto período de tiempo, dificultan el diagnóstico. Por tanto, es importante no olvidarnos de esta patología ante un paciente con espasmos musculares y ausencia o historia incompleta de vacunación.

  8. The complete genome sequence of Clostridium indolis DSM 755(T.).

    Science.gov (United States)

    Biddle, Amy S; Leschine, Susan; Huntemann, Marcel; Han, James; Chen, Amy; Kyrpides, Nikos; Markowitz, Victor; Palaniappan, Krishna; Ivanova, Natalia; Mikhailova, Natalia; Ovchinnikova, Galina; Schaumberg, Andrew; Pati, Amrita; Stamatis, Dimitrios; Reddy, Tatiparthi; Lobos, Elizabeth; Goodwin, Lynne; Nordberg, Henrik P; Cantor, Michael N; Hua, Susan X; Woyke, Tanja; Blanchard, Jeffrey L

    2014-06-15

    Clostridium indolis DSM 755(T) is a bacterium commonly found in soils and the feces of birds and mammals. Despite its prevalence, little is known about the ecology or physiology of this species. However, close relatives, C. saccharolyticum and C. hathewayi, have demonstrated interesting metabolic potentials related to plant degradation and human health. The genome of C. indolis DSM 755(T) reveals an abundance of genes in functional groups associated with the transport and utilization of carbohydrates, as well as citrate, lactate, and aromatics. Ecologically relevant gene clusters related to nitrogen fixation and a unique type of bacterial microcompartment, the CoAT BMC, are also detected. Our genome analysis suggests hypotheses to be tested in future culture based work to better understand the physiology of this poorly described species.

  9. Blowhole Colostomy for Clostridium difficile-Associated Toxic Megacolon

    Directory of Open Access Journals (Sweden)

    Jeroen Kerstens

    2016-01-01

    Full Text Available We present the case of a 58-year-old man who underwent urgent blowhole colostomy for toxic megacolon (TM secondary to Clostridium difficile infection (CDI. This infection occurred under antibiotic coverage with amoxicillin-clavulanic acid, four days after laparoscopic sigmoidectomy in our hospital. Although prospective clinical research regarding the surgical management of TM is lacking, decompressive procedures like blowhole colostomy are reported to carry a high risk of postoperative morbidity and mortality and are widely regarded as obsolete. Subtotal or total colectomy with end ileostomy is currently considered the procedure of choice. After presenting our case, we discuss the literature available on the subject to argue that the scarce evidence on the optimal surgical treatment for TM is primarily based on TM associated with inflammatory bowel diseases (IBD and that there might be a rationale for considering minimally invasive procedures like blowhole colostomy for CDI-associated TM.

  10. Stimulation of Clostridium perfringens enterotoxin formation by caffeine and theobromine.

    Science.gov (United States)

    Labbe, R G; Nolan, L L

    1981-01-01

    In the presence of 100 micrograms of caffeine per ml or 200 micrograms of theobromine per ml, sporulation of Clostridium perfringens NCTC 8679 rose from less than 1 to 80 or 85%. Enterotoxin concentration increased from undetectable levels to 450 micrograms/mg of cell extract protein. Heat-resistant spore levels increased from less than 1,000 to between 1 X 10(7) and 2 X 10(7)/ml. These effects were partially reversible by the addition of adenosine or thymidine. In the case of NCTC 8238, caffeine and theobromine caused a three- to fourfold increase in the percentages of cells possessing refractile spores and a similar increase in enterotoxin concentration. Heat-resistant spore levels, however, were unaffected. Inosine was ineffective in promoting sporulation in NCTC 8679. PMID:6271685

  11. Rectal bacteriotherapy for recurrent Clostridium difficile-associated diarrhoea

    DEFF Research Database (Denmark)

    Tvede, M; Tinggaard, M; Helms, M

    2015-01-01

    Clostridium difficile infection is one of the most common nosocomial infections. Among other alternatives to standard treatment with vancomycin for recurrent infection are faecal microbiota transplantation and rectal bacteriotherapy with a fixed mixture of intestinal bacterial strains isolated from...... for relapsing C. difficile in Denmark, 2000-2012. The primary end point was recurrent diarrhoea within 30 days after treatment. A total of 55 patients were included in this case series. Thirty-five patients (64%) had no recurrence within 30 days of bacteriotherapy. Patients with recurrence tended to be older....... difficile episode less than 6 months before bacteriotherapy. The most common adverse events were abdominal pain (10.9%) and worsening diarrhoea (4.3%). One patient was hospitalized 10 days after treatment with appendicitis, fever, and Escherichia coli bacteremia. The results from this study indicate...

  12. Clostridium difficile infection: Evolution, phylogeny and molecular epidemiology.

    Science.gov (United States)

    Elliott, Briony; Androga, Grace O; Knight, Daniel R; Riley, Thomas V

    2017-04-01

    Over the recent decades, Clostridium difficile infection (CDI) has emerged as a global public health threat. Despite growing attention, C. difficile remains a poorly understood pathogen, however, the exquisite sensitivity offered by next generation sequencing (NGS) technology has enabled analysis of the genome of C. difficile, giving us access to massive genomic data on factors such as virulence, evolution, and genetic relatedness within C. difficile groups. NGS has also demonstrated excellence in investigations of outbreaks and disease transmission, in both small and large-scale applications. This review summarizes the molecular epidemiology, evolution, and phylogeny of C. difficile, one of the most important pathogens worldwide in the current antibiotic resistance era. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Optimizing the diagnostic testing of Clostridium difficile infection.

    Science.gov (United States)

    Bouza, Emilio; Alcalá, Luis; Reigadas, Elena

    2016-09-01

    Clostridium difficile infection (CDI) is the leading cause of hospital-acquired diarrhea and is associated with a considerable health and cost burden. However, there is still not a clear consensus on the best laboratory diagnosis approach and a wide variation of testing methods and strategies can be encountered. We aim to review the most practical aspects of CDI diagnosis providing our own view on how to optimize CDI diagnosis. Expert commentary: Laboratory diagnosis in search of C. difficile toxins should be applied to all fecal diarrheic samples reaching the microbiology laboratory in patients > 2 years old, with or without classic risk factors for CDI. Detection of toxins either directly in the fecal sample or in the bacteria isolated in culture confirm CDI in the proper clinical setting. Nuclear Acid Assay techniques (NAAT) allow to speed up the process with epidemiological and therapeutic consequences.

  14. Asymptomatic carriers contribute to nosocomial Clostridium difficile infection

    DEFF Research Database (Denmark)

    Blixt, Thomas; Gradel, Kim Oren; Homann, Christian

    2017-01-01

    BACKGROUND & AIMS: Nosocomial infection with Clostridium difficile pose a considerable problem despite numerous attempts by health care workers to reduce risk of transmission. Asymptomatic carriers of C difficile might spread their infection to other patients. We investigated the effects...... of of asymptomatic carriers on nosocomial C difficile infections. METHODS: We performed a population-based prospective cohort study at 2 university hospitals in Denmark, screening all patients for toxigenic C difficile in the intestine upon admittance, from October 1, 2012, to January 31, 2013. Screening results...... were blinded to patients, staff, and researchers. Patients were followed during their hospital stay by daily registration of wards and patient rooms. The primary outcomes were rate of C difficile infection in exposed and unexposed patients and factors associated with transmission. RESULTS: C difficile...

  15. Clostridium difficile infection: molecular pathogenesis and novel therapeutics

    Science.gov (United States)

    Rineh, Ardeshir; Kelso, Michael J; Vatansever, Fatma; Tegos, George P; Hamblin, Michael R

    2015-01-01

    The Gram-positive anaerobic bacterium Clostridium difficile produces toxins A and B, which can cause a spectrum of diseases from pseudomembranous colitis to C. difficile-associated diarrhea. A limited number of C. difficile strains also produce a binary toxin that exhibits ADP ribosyltransferase activity. Here, the structure and the mechanism of action of these toxins as well as their role in disease are reviewed. Nosocomial C. difficile infection is often contracted in hospital when patients treated with antibiotics suffer a disturbance in normal gut microflora. C. difficile spores can persist on dry, inanimate surface for months. Metronidazole and oral vancomycin are clinically used for treatment of C. difficile infection but clinical failure and concern about promotion of resistance are motivating the search for novel non-antibiotic therapeutics. Methods for controlling both toxins and spores, replacing gut microflora by probiotics or fecal transplant, and killing bacteria in the anaerobic gut by photodynamic therapy are discussed. PMID:24410618

  16. Rethinking Strategies to Select Antibiotic Therapy in Clostridium difficile infection.

    Science.gov (United States)

    Chopra, Teena; Goldstein, Ellie J C; Gorbach, Sherwood L

    2016-12-01

    In recent years, Clostridium difficile infection (CDI) has become a global public health threat associated with increased morbidity, mortality, and economic burden, all of which are exacerbated with disease recurrence. Current guidelines informing treatment decisions are largely based on definitions of disease severity at diagnosis, with subjective components not well delineated across treatment algorithms and clinical trials. Furthermore, there is little evidence linking severity at onset to outcome. However, reducing the risk of recurrence may offer both a better outcome for the individual and decreased downstream economic impact. The authors present data supporting the opinion that patients deemed at low risk for recurrence should receive vancomycin (or metronidazole when cost is an issue), while those at higher risk of recurrence would benefit from fidaxomicin treatment. Although further prospective studies are needed, choosing treatment with the goal of preventing recurrent CDI may offer a better guide than disease severity. © 2016 Pharmacotherapy Publications, Inc.

  17. Pomegranate extract exhibits in vitro activity against Clostridium difficile.

    Science.gov (United States)

    Finegold, Sydney M; Summanen, Paula H; Corbett, Karen; Downes, Julia; Henning, Susanne M; Li, Zhaoping

    2014-10-01

    To determine the possible utility of pomegranate extract in the management or prevention of Clostridium difficile infections or colonization. The activity of pomegranate was tested against 29 clinical C. difficile isolates using the Clinical and Laboratory Standards Institute-approved agar dilution technique. Total phenolics content of the pomegranate extract was determined by Folin-Ciocalteau colorimetric method and final concentrations of 6.25 to 400 μg/mL gallic acid equivalent were achieved in the agar. All strains had MICs at 12.5 to 25 mg/mL gallic acid equivalent range. Our results suggest antimicrobial in vitro activity for pomegranate extract against toxigenic C. difficile. Pomegranate extract may be a useful contributor to the management and prevention of C. difficile disease or colonization. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Molecular diversity of Clostridium botulinum and phenotypically similar strains.

    Science.gov (United States)

    Grenda, T; Kukier, E; Sieradzki, Z; Goldsztejn, M; Kwiatek, K

    2016-12-01

    This study was undertaken to examine phenotypic and genetic features of strains preliminary classified as Clostridium botulinum species. The phenotypic characteristics were assessed with different culture media and biochemical tests. The genetic characterization included detection of botulinum toxin genes by PCR and macrorestriction analysis with SmaI, XhoI and SacII by PFGE (Pulsed-field Gel Electrophoresis). Despite similar biochemical properties of all analysed strains, only 47% of them contained genes determining toxicity specific to C. botulinum species. The most valuable differentiation of C. botulinum and C. botulinum-like strains was obtained after SmaI digestion. The highest affinity was observed among C. botulinum type B profiles which was even up to 100%. It was found 100% of affinity between C. botulinum and C. botulinum-like strains, however, the similarity among C. botulinum and C. botulinum-like was generally lower than 80%.

  19. Recurrent Clostridium difficile Infection: From Colonization to Cure

    Science.gov (United States)

    Shields, Kelsey; Araujo-Castillo, Roger V.; Theethira, Thimmaiah G.; Alonso, Carolyn D.; Kelly, Ciaran

    2015-01-01

    Clostridium difficile infection (CDI) is increasingly prevalent, dangerous and challenging to prevent and manage. Despite intense national and international attention the incidence of primary and of recurrent CDI (PCDI and RCDI, respectively) have risen rapidly throughout the past decade. Of major concern is the increase in cases of RCDI resulting in substantial morbidity, morality and economic burden. RCDI management remains challenging as there is no uniformly effective therapy, no firm consensus on optimal treatment, and reliable data regarding RCDI-specific treatment options is scant. Novel therapeutic strategies are critically needed to rapidly, accurately, and effectively identify and treat patients with, or at-risk for, RCDI. In this review we consider the factors implicated in the epidemiology, pathogenesis and clinical presentation of RCDI, evaluate current management options for RCDI and explore novel and emerging therapies. PMID:25930686

  20. Recent Insights into Clostridium perfringens Beta-Toxin

    Directory of Open Access Journals (Sweden)

    Masahiro Nagahama

    2015-02-01

    Full Text Available 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.

  1. Global analysis of the sporulation pathway of Clostridium difficile.

    Science.gov (United States)

    Fimlaid, Kelly A; Bond, Jeffrey P; Schutz, Kristin C; Putnam, Emily E; Leung, Jacqueline M; Lawley, Trevor D; Shen, Aimee

    2013-01-01

    The Gram-positive, spore-forming pathogen Clostridium difficile is the leading definable cause of healthcare-associated diarrhea worldwide. C. difficile infections are difficult to treat because of their frequent recurrence, which can cause life-threatening complications such as pseudomembranous colitis. The spores of C. difficile are responsible for these high rates of recurrence, since they are the major transmissive form of the organism and resistant to antibiotics and many disinfectants. Despite the importance of spores to the pathogenesis of C. difficile, little is known about their composition or formation. Based on studies in Bacillus subtilis and other Clostridium spp., the sigma factors σ(F), σ(E), σ(G), and σ(K) are predicted to control the transcription of genes required for sporulation, although their specific functions vary depending on the organism. In order to determine the roles of σ(F), σ(E), σ(G), and σ(K) in regulating C. difficile sporulation, we generated loss-of-function mutations in genes encoding these sporulation sigma factors and performed RNA-Sequencing to identify specific sigma factor-dependent genes. This analysis identified 224 genes whose expression was collectively activated by sporulation sigma factors: 183 were σ(F)-dependent, 169 were σ(E)-dependent, 34 were σ(G)-dependent, and 31 were σ(K)-dependent. In contrast with B. subtilis, C. difficile σ(E) was dispensable for σ(G) activation, σ(G) was dispensable for σ(K) activation, and σ(F) was required for post-translationally activating σ(G). Collectively, these results provide the first genome-wide transcriptional analysis of genes induced by specific sporulation sigma factors in the Clostridia and highlight that diverse mechanisms regulate sporulation sigma factor activity in the Firmicutes.

  2. Attributable Cost of Clostridium difficile Infection in Pediatric Patients.

    Science.gov (United States)

    Mehrotra, Preeti; Jang, Jisun; Gidengil, Courtney; Sandora, Thomas J

    2017-12-01

    OBJECTIVES The attributable cost of Clostridium difficile infection (CDI) in children is unknown. We sought to determine a national estimate of attributable cost and length of stay (LOS) of CDI occurring during hospitalization in children. DESIGN AND METHODS We analyzed discharge records of patients between 2 and 18 years of age from the Agency for Healthcare Research and Quality (AHRQ) Kids' Inpatient Database. We created a logistic regression model to predict CDI during hospitalization based on demographic and clinical characteristics. Predicted probabilities from the logistic regression model were then used as propensity scores to match 1:2 CDI to non-CDI cases. Charges were converted to costs and compared between patients with CDI and propensity-score-matched controls. In a sensitivity analysis, we adjusted for LOS as a confounder by including it in both the propensity score and a generalized linear model predicting cost. RESULTS We identified 8,527 pediatric hospitalizations (0.53%) with a diagnosis of CDI and 1,597,513 discharges without CDI. In our matched cohorts, the attributable cost of CDI occurring during a hospitalization ranged from $1,917 to $8,317, depending on whether model was adjusted for LOS. When not adjusting for LOS, CDI-associated hospitalizations cost 1.6 times more than non-CDI associated hospitalizations. Attributable LOS of CDI was approximately 4 days. CONCLUSIONS Clostridium difficile infection in hospitalized children is associated with an economic burden similar to adult estimates. This finding supports a continued focus on preventing CDI in children as a priority. Pediatric CDI cost analyses should account for LOS as an important confounder of cost. Infect Control Hosp Epidemiol 2017;38:1472-1477.

  3. Incidence and Costs of Clostridium difficile Infections in Canada

    Science.gov (United States)

    Levy, Adrian R.; Szabo, Shelagh M.; Lozano-Ortega, Greta; Lloyd-Smith, Elisa; Leung, Victor; Lawrence, Robin; Romney, Marc G.

    2015-01-01

    Background. Limited data are available on direct medical costs and lost productivity due to Clostridium difficile infection (CDI) in Canada. Methods. We developed an economic model to estimate the costs of managing hospitalized and community-dwelling patients with CDI in Canada. The number of episodes was projected based on publicly available national rates of hospital-associated CDI and the estimate that 64% of all CDI is hospital-associated. Clostridium difficile infection recurrences were classified as relapses or reinfections. Resource utilization data came from published literature, clinician interviews, and Canadian CDI surveillance programs, and this included the following: hospital length of stay, contact with healthcare providers, pharmacotherapy, laboratory testing, and in-hospital procedures. Lost productivity was considered for those under 65 years of age, and the economic impact was quantified using publicly available labor statistics. Unit costs were obtained from published sources and presented in 2012 Canadian dollars. Results. There were an estimated 37 900 CDI episodes in Canada in 2012; 7980 (21%) of these were relapses, out of a total of 10 900 (27%) episodes of recurrence. The total cost to society of CDI was estimated at $281 million; 92% ($260 million) was in-hospital costs, 4% ($12 million) was direct medical costs in the community, and 4% ($10 million) was due to lost productivity. Management of CDI relapses alone accounted for $65.1 million (23%). Conclusions. The largest proportion of costs due to CDI in Canada arise from extra days of hospitalization. Interventions reducing the severity of infection and/or relapses leading to rehospitalizations are likely to have the largest absolute effect on direct medical costs. PMID:26191534

  4. Clostridium difficile infection in the elderly: an update on management

    Directory of Open Access Journals (Sweden)

    Asempa TE

    2017-10-01

    Full Text Available Tomefa E Asempa, David P Nicolau Center for Anti-Infective Research and Development, Hartford Hospital, Hartford, CT, USA Abstract: The burden of Clostridium difficile infection (CDI is profound and growing. CDI now represents a common cause of health care–associated diarrhea, and is associated with significant morbidity, mortality, and health care costs. CDI disproportionally affects the elderly, possibly explained by the following risk factors: age-related impairment of the immune system, increasing antibiotic utilization, and frequent health care exposure. In the USA, recent epidemiological studies estimate that two out of every three health care–associated CDIs occur in patients 65 years or older. Additionally, the elderly are at higher risk for recurrent CDI. Existing therapeutic options include metronidazole, oral vancomycin, and fidaxomicin. Choice of agent depends on disease severity, history of recurrence, and, increasingly, the drug cost. Bezlotoxumab, a recently approved monoclonal antibody targeting C. difficile toxin B, offers an exciting advancement into immunologic therapies. Similarly, fecal microbiota transplantation is gaining popularity as an effective option mainly for recurrent CDI. The challenge of decreasing CDI burden in the elderly involves adopting preventative strategies, optimizing initial treatment, and decreasing the risk of recurrence. Expanded strategies are certainly needed to improve outcomes in this high-risk population. This review considers available data from prospective and retrospective studies as well as case reports to illustrate the merits and gaps in care related to the management of CDI in the elderly. Keywords: Clostridium difficile, recurrence, risk factors, elderly, aging, treatment, bezlotoxumab, fecal microbiota transplant

  5. Rheological properties of erythrocytes in patients infected with Clostridium difficile.

    Science.gov (United States)

    Czepiel, Jacek; Jurczyszyn, Artur; Biesiada, Grażyna; Sobczyk-Krupiarz, Iwona; Jałowiecka, Izabela; Świstek, Magdalena; Perucki, William; Teległów, Aneta; Marchewka, Jakub; Dąbrowski, Zbigniew; Mach, Tomasz; Garlicki, Aleksander

    2014-12-04

    Clostridium difficile infection (CDI) is a bacterial infection of the digestive tract. Acute infections are accompanied by increased risk for venous thromboembolism (VTE). To date, there have been no studies of the rheological properties of blood during the course of digestive tract infections. The aim of our study was to examine the effects of CDI on red blood cell (RBC) rheology, specifically RBC deformability, RBC aggregation, and plasma viscosity. In addition, the activity of glucose 6 phosphate dehydrogenase (G6PD) and acetylcholinesterase (AChE) in RBC was studied. Our study group included 20 patients with CDI, 20 healthy persons comprised the control group. We examined the effects of CDI on the rheology of RBCs, their deformability and aggregation, using a Laser-assisted Optical Rotational Cell Analyzer (LORCA). Plasma viscosity was determined using a capillary tube plasma viscosymeter. Moreover, we estimated the activity of AChE and G6PD in RBC using spectrophotometric method. A statistically significant increase was found in the aggregation index, viscosity and activity of G6PD whereas the amount of time to reach half of maximum aggregation (t½) and the amplitude of aggregation (AMP) both showed statistically significantly decreases among patients with CDI compared to the control group. We also observed that the Elongation Index (EI) was decreased when shear stress values were low, between 0.3 Pa and 0.58 Pa, whereas EI was increased for shear stress in the range of 1.13-59.97 Pa. These observations were statistically significant. We report for the first time that acute infection of the gastrointestinal tract with Clostridium difficile is associated with abnormalities in rheological properties of blood, increased serum viscosity as well as increased aggregation of RBCs, which correlated with severity of inflammation. These abnormalities may be an additional mechanism causing increased incidence of VTE in CDI.

  6. Rheological properties of erythrocytes in patients infected with Clostridium difficile

    Directory of Open Access Journals (Sweden)

    Jacek Czepiel

    2014-12-01

    Full Text Available Clostridium difficile infection (CDI is a bacterial infection of the digestive tract. Acute infections are accompanied by increased risk for venous thromboembolism (VTE. To date, there have been no studies of the rheological properties of blood during the course of digestive tract infections. The aim of our study was to examine the effects of CDI on red blood cell (RBC rheology, specifically RBC deformability, RBC aggregation, and plasma viscosity. In addition, the activity of glucose 6 phosphate dehydrogenase (G6PD and acetylcholinesterase (AChE in RBC was studied. Our study group included 20 patients with CDI, 20 healthy persons comprised the control group. We examined the effects of CDI on the rheology of RBCs, their deformability and aggregation, using a Laser–assisted Optical Rotational Cell Analyzer (LORCA. Plasma viscosity was determined using a capillary tube plasma viscosymeter. Moreover, we estimated the activity of AChE and G6PD in RBC using spectrophotometric method. A statistically significant increase was found in the aggregation index, viscosity and activity of G6PD whereas the amount of time to reach half of maximum aggregation (t½ and the amplitude of aggregation (AMP both showed statistically significantly decreases among patients with CDI compared to the control group. We also observed that the Elongation Index (EI was decreased when shear stress values were low, between 0.3 Pa and 0.58 Pa, whereas EI was increased for shear stress in the range of 1.13 - 59.97 Pa. These observations were statistically significant. We report for the first time that acute infection of the gastrointestinal tract with Clostridium difficile is associated with abnormalities in rheological properties of blood, increased serum viscosity as well as increased aggregation of RBCs, which correlated with severity of inflammation. These abnormalities may be an additional mechanism causing increased incidence of VTE in CDI.

  7. Deuterium nuclear magnetic resonance studies on the plasmalogens and the glycerol acetals of plasmalogens of Clostridium butyricum and Clostridium beijerinckii

    International Nuclear Information System (INIS)

    Malthaner, M.; Seelig, J.; Johnston, N.C.; Goldfine, H.

    1987-01-01

    Deuterium nuclear magnetic resonance was used to investigate the structure of different lipid fractions isolated from the anaerobic bacteria Clostridium butyricum and Clostridium beijerinckii. The fractions isolated from C. butyricum were (1) phosphatidylethanolamine/plasmenylethanolamine and (2) the glycerol acetal of plasmenylethanolamine, and from C. beijerinckii similar fractions containing principally (1) phosphatidyl-N-monomethylethanolamine, along with its plasmalogen, and (2) the glycerol acetal of this plasmalogen were isolated. The third fraction from both species consisted largely of the acidic lipids phosphatidylglycerol and cardiolipin along with plasmalogen forms of these lipids. Palmitic acid with deuterium labels at C-2, C-3, or C-4 or oleic acid with deuterium labels at C-2 and C-9,10 was added to the growth medium and incorporated to various extents in the lipid fractions. Biochemical analysis showed that palmitic acid and oleic acid were preferentially bound to the sn-2 and sn-1 positions, respectively, of the glycerol backbone when both fatty acids were added to the medium. From the 2 H NMR spectra, the hydrocarbon chain ordering near the lipid-water interface could be determined and appeared to be similar for all three lipid fractions. The deuterium quadrupole splitting and order parameter were low at the C-2 segment and increased by almost a factor of 2 at positions C-3 and C-4 for cells fed with deuteriated palmitic acid along with unlabeled oleic acid. These results agree with previous findings on pure diacyl lipids in which the sn-2 chain was found to adopt a bent conformation at the carbon segment C-2. However, two unusual quadrupole splittings could be detected for the plasmalogens. By comparison with other model systems it could be concluded that the double bond is aligned essentially parallel with the long axis of the hydrocarbon chains

  8. Use of natural ingredients to control growth of Clostridium perfringens in naturally cured frankfurters and hams.

    Science.gov (United States)

    Jackson, Armitra L; Kulchaiyawat, Charlwit; Sullivan, Gary A; Sebranek, Joseph G; Dickson, James S

    2011-03-01

    A major concern for processed meats marketed as natural/organic is that they do not contain nitrite in concentrations known to be most effective for inhibiting foodborne pathogens. Supplemental treatments to increase the level and consistency of antimicrobial protection in these products may be important to provide consumers with the degree of safety that they have come to expect from conventionally cured meats. Therefore, the objective of this study was to identify and test ingredients that might improve processed meat product safety without altering their natural/organic status. Eight treatments of hams and frankfurters were prepared: (A) uncured control (typical ingredients except nitrite and nitrate); (B) conventionally cured control (erythorbate, nitrite, and a lactate-diacetate blend); (C) natural nitrate cure (including starter culture containing Staphylococcus carnosus); (D) natural nitrate cure (culture and natural antimicrobial A containing a vinegar, lemon, and cherry powder blend); (E) natural nitrate cure (culture and antimicrobial B containing a cultured sugar and vinegar blend); (F) natural nitrite cure without additional antimicrobials; (G) natural nitrite cure with natural antimicrobial A; and (H) natural nitrite cure with antimicrobial B. For the hams, treatments C, D, E, and H impacted growth of Clostridium perfringens to the same extent (P cured control (approximately 2 log less growth over time than uncured control). For frankfurters, treatments D, G, and H had an effect (approximately 1 log) on growth equivalent to that of the conventionally cured control (P cured meats have more potential for pathogen growth than conventionally cured products, but supplemental natural ingredients offer safety improvement.

  9. Effects of gamma radiation on Clostridium botulinum type E under various parameters

    International Nuclear Information System (INIS)

    Lim, Y.H.

    1986-01-01

    Spores of Clostridium botulinum type E strain Eklund (Eklund) was irradiated with gamma radiation and its recovery was tested on the tryptone-peptone-glucose-yeast extract-agar (TPGYA) containing various levels of NaCl and Na-thioglycollate. The presence of 0.5% or more NaCl in the media decreased the viable counts, while Na-thioglycollate of up to 0.15% did not affect the recovery of both irradiated and non-irradiated spores. Eklund spores were also irradiated under air (21% O 2 ), N 2 O and N 2 , with or without the additive of one of the following agents (additive/concentration): disodium ethylenediaminetetraacetate (EDTA), 0.01 M; t-butanol, 0.1 M; NaCl, 0.01 M; catalyze, 10 mg/ml and DL-cysteine, 0.1 mM. Radiation process was most effective in destroying the spores when carried out under air (21% O 2 ), followed by N 2 O and N 2 . Among the additives tested, EDTA was the most efficient protector followed by t-butanol when irradiation process was carried under N 2 O and N 2 gas environment. Catalase and DL-cysteine sensitized the spores when irradiated under N 2 O and N 2 , while NaCl only sensitized under N 2 . Spores kept frozen at -75 0 C for 30 days but thawed prior to irradiation were more sensitive to radiation damage than freshly prepared spores. Radiation resistance of the spores increased when 15% glycerol was added to the phosphate bugger (0.06 M, pH 7.0) and used as suspending media. When the concentration of the spore increased from 10 6 /ml to 10 11 /ml, the radiosensitivities also increased. Seven strains of C. botulinum type E were screened for plasmids by agarose gel electrophoresis

  10. Clostridium Perfringens Epsilon Toxin Binds to Membrane Lipids and Its Cytotoxic Action Depends on Sulfatide.

    Directory of Open Access Journals (Sweden)

    Carles Gil

    Full Text Available Epsilon toxin (Etx is one of the major lethal toxins produced by Clostridium perfringens types B and D, being the causal agent of fatal enterotoxemia in animals, mainly sheep and goats. Etx is synthesized as a non-active prototoxin form (proEtx that becomes active upon proteolytic activation. Etx exhibits a cytotoxic effect through the formation of a pore in the plasma membrane of selected cell targets where Etx specifically binds due to the presence of specific receptors. However, the identity and nature of host receptors of Etx remain a matter of controversy. In the present study, the interactions between Etx and membrane lipids from the synaptosome-enriched fraction from rat brain (P2 fraction and MDCK cell plasma membrane preparations were analyzed. Our findings show that both Etx and proEtx bind to lipids extracted from lipid rafts from the two different models as assessed by protein-lipid overlay assay. Lipid rafts are membrane microdomains enriched in cholesterol and sphingolipids. Binding of proEtx to sulfatide, phosphatidylserine, phosphatidylinositol (3-phosphate and phosphatidylinositol (5-phosphate was detected. Removal of the sulphate groups via sulfatase treatment led to a dramatic decrease in Etx-induced cytotoxicity, but not in proEtx-GFP binding to MDCK cells or a significant shift in oligomer formation, pointing to a role of sulfatide in pore formation in rafts but not in toxin binding to the target cell membrane. These results show for the first time the interaction between Etx and membrane lipids from host tissue and point to a major role for sulfatides in C. perfringens epsilon toxin pathophysiology.

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

    Energy Technology Data Exchange (ETDEWEB)

    Banerjee, A; Leang, C; Ueki, T; Nevin, KP; Lovley, DR

    2014-03-25

    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.

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

    Science.gov (United States)

    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

  13. Treatment of relapsing Clostridium difficile infection using fecal microbiota transplantation

    Directory of Open Access Journals (Sweden)

    Pathak R

    2013-12-01

    Full Text Available Rahul Pathak,1 Hill Ambrose Enuh,1 Anish Patel,1 Prasanna Wickremesinghe21Department of Internal Medicine, New York Medical College, Internal Medicine Program at Richmond University Medical Center, Staten Island, NY, USA; 2Department of Gastrointestinal Medicine, New York Medical College, Internal Medicine Program at Richmond University Medical Center, Staten Island, NY, USABackground: Clostridium difficile infection (CDI has become a global concern over the last decade. In the United States, CDI escalated in incidence from 1996 to 2005 from 31 to 64/100,000. In 2010, there were 500,000 cases of CDI with an estimated mortality up to 20,000 cases a year. The significance of this problem is evident from the hospital costs of over 3 billion dollars annually. Fecal microbiota transplant (FMT was first described in 1958 and since then about 500 cases have been published in literature in various small series and case reports. This procedure has been reported mainly from centers outside of the United States and acceptance of the practice has been difficult. Recently the US Food and Drug Administration (FDA labeled FMT as a biological drug; as a result, guidelines will soon be required to help establish it as a mainstream treatment. More US experience needs to be reported to popularize this procedure here and form guidelines.Method: We did a retrospective review of our series of patients with relapsing CDI who were treated with FMT over a 3-year period. We present our experience with FMT at a community hospital as a retrospective review and describe our procedure.Results: There were a total of 12 patients who underwent FMT for relapsing C. difficile. Only one patient failed to respond and required a second FMT. There were no complications associated with the transplant and all patients had resolution of symptoms within 48 hours of FMT.Conclusion: FMT is a cheap, easily available, effective therapy for recurrent CDI; it can be safely performed in a

  14. The effect of Clostridium difficile infection on cardiac surgery outcomes.

    Science.gov (United States)

    Lemaire, Anthony; Dombrovskiy, Viktor; Batsides, George; Scholz, Peter; Solina, Al; Brownstone, Nicholas; Spotnitz, Alan; Lee, Leonard Y

    2015-02-01

    Clostridium difficile (CD) is a common cause of healthcare-associated infectious colitis that complicates about 1% of all hospital stays in the U.S. The impact of CD on outcomes after coronary artery bypass grafting (CABG) and valvular surgery (VS) is not well known. The Nationwide Inpatient Sample (2002-2009) was queried to identify CABG and VS patients utilizing International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) diagnosis codes. Rates of CD, post-operative endocarditis and mediastinitis, hospital mortality rate, and resource utilization were evaluated. We identified 421,294 and 90,923 patients of age 40 yrs and older who underwent CABG and VS, respectively. The CD infection was more likely to develop in patients undergoing VS than in those having CABG (odds ratio [OR] 1.8; 95% confidence interval [CI] 1.64-1.92) and was more likely after urgent or emergency admission than after elective admission (OR 1.8; 95% CI 1.68-1.94). There was a greater likelihood of mediastinitis in patients with CD after CABG than in non-complicated cases without CD, both by univariable (OR 6.0; 95% CI 3.07-11.62) and multivariable analysis with adjustment for patient age, gender, race, type of admission, and co-morbidities (OR 3.1; 95% CI 1.49-6.51). The infection thus was most likely a result of the antibiotics used to treat mediastinitis, as the patients treated for mediastinitis were most likely to develop CD. There was a significant association in patients with CD and endocarditis who underwent VS but not in patients who did not have CD. The CD infection in these patients thus was most likely a result of the antibiotics used to treat endocarditis. Endocarditis and CD developed 3.2 times (95% CI 2.65-3.97) as often as in patients without CD, a finding that was confirmed by multivariable analysis (OR 2.2; 95% CI 1.70-2.84). At the same time, in patients having VS, there was no significant association of CD and mediastinitis. Clostridium

  15. Effect of Cultured Celery Juice, Temperature, and Product Composition on the Inhibition of Proteolytic Clostridium botulinum Toxin Production.

    Science.gov (United States)

    Golden, Max C; Wanless, Brandon J; David, Jairus R D; Kottapalli, Bala; Lineback, D Scott; Talley, Ryan J; Glass, Kathleen A

    2017-08-01

    Clostridium botulinum may be of concern in prepared refrigerated meals, for which strict cold chain management cannot be guaranteed. This study evaluated the effect of temperature, product composition, and cultured celery juice powder (CCJP) as a source of nitrite on the inhibition of botulinum toxin formation in two experimental (meat- and vegetable-based) prepared meals. Data obtained from the challenge study were compared with a published mathematical model to determine whether the model is fail-safe with regard to the tested meals. Treatments were inoculated with proteolytic C. botulinum, vacuum packaged, cooked at 90°C for 10 min, and assayed for botulinum toxin at appropriate intervals in samples stored at 10, 15, or 20°C for up to 8 weeks. None of the treatments stored at 10°C for 8 weeks supported toxin production by proteolytic C. botulinum. The addition of CCJP delayed toxin production by 1 and 3 weeks in cauliflower potatoes and in Dijon pork, respectively, stored at 15°C. Toxin production was delayed by 1 week at 20°C when CCJP was added to the cauliflower potatoes. This study found that the predictive model was fail-safe but was overly conservative for the experimental meals described. Finally, this study confirms that product composition, the addition of nitrite via CCJP, storage time, and temperature play important roles in the inhibition of toxin formation by proteolytic C. botulinum.

  16. Clostridium difficile toxin CDT induces formation of microtubule-based protrusions and increases adherence of bacteria.

    Directory of Open Access Journals (Sweden)

    Carsten Schwan

    2009-10-01

    Full Text Available Clostridium difficile causes antibiotic-associated diarrhea and pseudomembranous colitis by production of the Rho GTPase-glucosylating toxins A and B. Recently emerging hypervirulent Clostridium difficile strains additionally produce the binary ADP-ribosyltransferase toxin CDT (Clostridium difficile transferase, which ADP-ribosylates actin and inhibits actin polymerization. Thus far, the role of CDT as a virulence factor is not understood. Here we report by using time-lapse- and immunofluorescence microscopy that CDT and other binary actin-ADP-ribosylating toxins, including Clostridium botulinum C2 toxin and Clostridium perfringens iota toxin, induce redistribution of microtubules and formation of long (up to >150 microm microtubule-based protrusions at the surface of intestinal epithelial cells. The toxins increase the length of decoration of microtubule plus-ends by EB1/3, CLIP-170 and CLIP-115 proteins and cause redistribution of the capture proteins CLASP2 and ACF7 from microtubules at the cell cortex into the cell interior. The CDT-induced microtubule protrusions form a dense meshwork at the cell surface, which wrap and embed bacterial cells, thereby largely increasing the adherence of Clostridia. The study describes a novel type of microtubule structure caused by less efficient microtubule capture and offers a new perspective for the pathogenetic role of CDT and other binary actin-ADP-ribosylating toxins in host-pathogen interactions.

  17. Butyric acid production from red algae by a newly isolated Clostridium sp. S1.

    Science.gov (United States)

    Lee, Kyung Min; Choi, Okkyoung; Kim, Ki-Yeon; Woo, Han Min; Kim, Yunje; Han, Sung Ok; Sang, Byoung-In; Um, Youngsoon

    2015-09-01

    To produce butyric acid from red algae such as Gelidium amansii in which galactose is a main carbohydrate, microorganisms utilizing galactose and tolerating inhibitors in hydrolysis including levulinic acid and 5-hydroxymethylfurfural (HMF) are required. A newly isolated bacterium, Clostridium sp. S1 produced butyric acid not only from galactose as the sole carbon source but also from a mixture of galactose and glucose through simultaneous utilization. Notably, Clostridium sp. S1 produced butyric acid and a small amount of acetic acid with the butyrate:acetate ratio of 45.4:1 and it even converted acetate to butyric acid. Clostridium sp. S1 tolerated 0.5-2 g levulinic acid/l and recovered from HMF inhibition at 0.6-2.5 g/l, resulting in 85-92% butyric acid concentration of the control culture. When acid-pretreated G. amansii hydrolysate was used, Clostridium sp. S1 produced 4.83 g butyric acid/l from 10 g galactose/l and 1 g glucose/l. Clostridium sp. S1 produces butyric acid from red algae due to its characteristics in sugar utilization and tolerance to inhibitors, demonstrating its advantage as a red algae-utilizing microorganism.

  18. Detection of Clostridium tyrobutyricum using cultivation and biochemical methods and polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    Radka Burdychová

    2007-01-01

    Full Text Available Anaerobic spore-forming bacteria of the genus Clostridium are commonly present in raw milk and some milk products. Their spores can survive pasteurization and can provoke so called late blowing defect in cheese caused by butyric acid fermentation. The only species of the genus Clostridium that is able to provoke late blowing is Clostridium tyrobutyricum.In this work, two cultivation methods for detection of butyric acid producing clostridia in raw and pasteurized milk and in cheese samples were compared. The results show that tube method is suitable for route identification (in concentration 102 CFU/ml or /g of clostridia in milk and cheese. The standard cultivation technique is suitable for more sensitive identification (10 CFU/ml or /g. All presumptive colonies grown anaerobically on selective RCM agar with polymyxine B (500 μg/ml were classified to be of species Clostridium tyrobutyricum using PCR only. The confirmation using API tests were different in 50 % cases. The results show, that described PCR method is suitable for rapid screening of the presence of Clostridium tyrobutyricum in milk and cheese. PCR from one colony is possible to use for the analysis.

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

    Science.gov (United States)

    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.

  20. Prevalence and risk factors of Clostridium difficile infection in patients hospitalized for flare of inflammatory bowel disease: a retrospective assessment.

    Science.gov (United States)

    Regnault, Helene; Bourrier, Anne; Lalande, Valerie; Nion-Larmurier, Isabelle; Sokol, Harry; Seksik, Philippe; Barbut, Frederic; Cosnes, Jacques; Beaugerie, Laurent

    2014-12-01

    Recent studies have identified a high frequency of Clostridium difficile infections in patients with active inflammatory bowel disease. To retrospectively assess the determinants and results of Clostridium difficile testing upon the admission of patients hospitalized with active inflammatory bowel disease in a tertiary care centre and to determine the predicting factors of Clostridium difficile infections. We reviewed all admissions from January 2008 and December 2010 for inflammatory bowel disease flare-ups. A toxigenic culture and a stool cytotoxicity assay were performed for all patients tested for Clostridium difficile. Out of 813 consecutive stays, Clostridium difficile diagnostic assays have been performed in 59% of inpatients. The independent predictive factors for the testing were IBD (ulcerative colitis: OR 2.0, 95% CI 1.5-2.9; pClostridium difficile infection was present in 7.0% of the inpatients who underwent testing. In a multivariate analysis, the only independent predictor was the intake of nonsteroidal anti-inflammatory drugs within the two months before admission (OR 3.8, 95% CI 1.2-12.3; p=0.02). Clostridium difficile infection is frequently associated with active inflammatory bowel disease. Our study suggests that a recent intake of nonsteroidal anti-inflammatory drugs is a risk factor for inflammatory bowel disease -associated Clostridium difficile infection. Copyright © 2014 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

  1. Clostridium perfringens challenge and dietary fat type modifies performance, microbiota composition and histomorphology of the broiler chicken gastrointestinal tract

    DEFF Research Database (Denmark)

    Josefiak, Damian; Swiatkiewicz, S; Kieronczyk, B

    2016-01-01

    Belastung mit Clostridium perfringens und Futterfettquelle modifizieren die Leistung, die Zusammensetzung der Microbiota und die Histomorphologie des Verdauungstraktes beim Broiler......Belastung mit Clostridium perfringens und Futterfettquelle modifizieren die Leistung, die Zusammensetzung der Microbiota und die Histomorphologie des Verdauungstraktes beim Broiler...

  2. Personnel Preparation.

    Science.gov (United States)

    Fair, George, Ed.; Stodden, Robert, Ed.

    1981-01-01

    Three articles comprise a section on personnel preparation in vocational education. Articles deal with two inservice programs in career/vocational education for the handicapped and a project to train paraprofessionals to assist special educators in vocational education. (CL)

  3. Atmospheric vs. anaerobic processing of metabolome samples for the metabolite profiling of a strict anaerobic bacterium, Clostridium acetobutylicum.

    Science.gov (United States)

    Lee, Sang-Hyun; Kim, Sooah; Kwon, Min-A; Jung, Young Hoon; Shin, Yong-An; Kim, Kyoung Heon

    2014-12-01

    Well-established metabolome sample preparation is a prerequisite for reliable metabolomic data. For metabolome sampling of a Gram-positive strict anaerobe, Clostridium acetobutylicum, fast filtration and metabolite extraction with acetonitrile/methanol/water (2:2:1, v/v) at -20°C under anaerobic conditions has been commonly used. This anaerobic metabolite processing method is laborious and time-consuming since it is conducted in an anaerobic chamber. Also, there have not been any systematic method evaluation and development of metabolome sample preparation for strict anaerobes and Gram-positive bacteria. In this study, metabolome sampling and extraction methods were rigorously evaluated and optimized for C. acetobutylicum by using gas chromatography/time-of-flight mass spectrometry-based metabolomics, in which a total of 116 metabolites were identified. When comparing the atmospheric (i.e., in air) and anaerobic (i.e., in an anaerobic chamber) processing of metabolome sample preparation, there was no significant difference in the quality and quantity of the metabolomic data. For metabolite extraction, pure methanol at -20°C was a better solvent than acetonitrile/methanol/water (2:2:1, v/v/v) at -20°C that is frequently used for C. acetobutylicum, and metabolite profiles were significantly different depending on extraction solvents. This is the first evaluation of metabolite sample preparation under aerobic processing conditions for an anaerobe. This method could be applied conveniently, efficiently, and reliably to metabolome analysis for strict anaerobes in air. © 2014 Wiley Periodicals, Inc.

  4. Butanol production from thin stillage using Clostridium pasteurianum.

    Science.gov (United States)

    Ahn, Jae-Hyung; Sang, Byoung-In; Um, Youngsoon

    2011-04-01

    The production of butanol from thin stillage by Clostridium pasteurianum DSM 525 was evaluated in the paper. At initial pH values ranging from 5.0 to 7.0 C. pasteurianum DSM 525 produced 6.2-7.2 g/L of butanol utilizing glycerol in thin stillage as the main carbon source, with yields of 0.32-0.44 g butanol produced/g glycerol consumed, which are higher than previously reported yields (e.g., 0.14-0.31 g butanol/g glycerol, Biebl, 2001). Lactic acid in the thin stillage acted as a buffering agent, maintaining the pH of the medium within a range of 5.7-6.1. Lactic acid was also utilized along with glycerol, enhancing butanol production (6.5 g/L butanol vs. 8.7 g/L butanol with 0 and 16 g/L lactic acid, respectively). These results demonstrate the feasibility of cost-effective butanol production using thin stillage as a nutrient-containing medium with a pH buffering capacity. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Rebalancing Redox to Improve Biobutanol Production by Clostridium tyrobutyricum

    Directory of Open Access Journals (Sweden)

    Chao Ma

    2015-12-01

    Full Text Available Biobutanol is a sustainable green biofuel that can substitute for gasoline. Carbon flux has been redistributed in Clostridium tyrobutyricum via metabolic cell engineering to produce biobutanol. However, the lack of reducing power hampered the further improvement of butanol production. The objective of this study was to improve butanol production by rebalancing redox. Firstly, a metabolically-engineered mutant CTC-fdh-adhE2 was constructed by introducing heterologous formate dehydrogenase (fdh and bifunctional aldehyde/alcohol dehydrogenase (adhE2 simultaneously into wild-type C. tyrobutyricum. The mutant evaluation indicated that the fdh-catalyzed NADH-producing pathway improved butanol titer by 2.15-fold in the serum bottle and 2.72-fold in the bioreactor. Secondly, the medium supplements that could shift metabolic flux to improve the production of butyrate or butanol were identified, including vanadate, acetamide, sodium formate, vitamin B12 and methyl viologen hydrate. Finally, the free-cell fermentation produced 12.34 g/L of butanol from glucose using the mutant CTC-fdh-adhE2, which was 3.88-fold higher than that produced by the control mutant CTC-adhE2. This study demonstrated that the redox engineering in C. tyrobutyricum could greatly increase butanol production.

  6. Clostridium perfringens Sporulation and Sporulation-Associated Toxin Production

    Science.gov (United States)

    Li, Jihong; Paredes-Sabja, Daniel; Sarker, Mahfuzur R.; McClane, Bruce A.

    2015-01-01

    The ability of Clostridium perfringens to form spores plays a key role during the transmission of this Gram-positive bacterium to cause disease. Of particular note, the spores produced by food poisoning strains are often exceptionally resistant to food environment stresses such as heat, cold and preservatives, which likely facilitates their survival in temperature-abused foods. The exceptional resistance properties of spores made by most type A food poisoning strains and some type C foodborne disease strains involves their production of a variant small acid soluble protein-4 that binds more tightly to spore DNA compared to the small acid soluble protein-4 made by most other C. perfringens strains. Sporulation and germination by C. perfringens and Bacillus spp. share both similarities and differences. Finally, sporulation is essential for production of C. perfringens enterotoxin, which is responsible for the symptoms of C. perfringens type A food poisoning, the second most common bacterial foodborne disease in the USA. During this foodborne disease, C. perfringens is ingested with food and then, using sporulation-specific alternate sigma factors, this bacterium sporulates and produces the enterotoxin in the intestines. PMID:27337447

  7. Comparative epidemiology of Clostridium difficile infection: England and the USA.

    Science.gov (United States)

    King, Alice; Mullish, Benjamin H; Williams, Horace R T; Aylin, Paul

    2017-10-01

    To examine whether there is an epidemiological difference between Clostridium difficile infection (CDI) inpatient populations in England and the United States. A cross-sectional study. National administrative inpatient discharge data from England (Hospital Episode Statistics) and the USA (National Inpatient Sample) in 2012. De-identifiable non-obstetric inpatient discharges from the national datasets were used to estimate national CDI incidence in the United States and England using ICD9-CM(008.45) and ICD10(A04.7) respectively. The rate of CDI was calculated per 100 000 population using national population estimates. Rate per 100 000 inpatient discharges was also calculated separated by primary and secondary diagnosis of CDI. Age, sex and Elixhauser comorbidities profiles were examined. The USA had a higher rate of CDI compared to England: 115.1/100 000 vs. 19.3/100 000 population (P USA (OR 1.20 95% CI [1.18,1.22] P USA compared to England apart from dementia, which was greater in England (9.63% vs. 1.25%, P USA was much higher than in England. Age and comorbidity profiles also differed between CDI patients in both countries. The reasons for this are likely multi-factorial but may reflect national infection control policy. © The Author 2017. Published by Oxford University Press in association with the International Society for Quality in Health Care. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

  8. Bezlotoxumab: A Review in Preventing Clostridium difficile Infection Recurrence.

    Science.gov (United States)

    Deeks, Emma D

    2017-10-01

    Bezlotoxumab (Zinplava™) is a fully human monoclonal antibody against Clostridium difficile toxin B indicated for the prevention of C. difficile infection (CDI) recurrence in patients with a high recurrence risk. It is the first agent approved for recurrence prevention and is administered as a single intravenous infusion in conjunction with standard-of-care (SoC) antibacterial treatment for CDI. In well-designed, placebo-controlled, phase 3 trials (MODIFY 1 and 2), a single infusion of bezlotoxumab, given in combination with SoC antibacterial therapy for CDI in adults, was effective in reducing CDI recurrence in the 12 weeks post-treatment, with this benefit being seen mainly in the patients at high recurrence risk. Bezlotoxumab did not impact the efficacy of the antibacterials being used to treat the CDI and, consistent with its benefits on CDI recurrence, appeared to reduce the need for subsequent antibacterials, thus minimizing further gut microbiota disruption. Longer term, there were no further CDI recurrences over 12 months' follow-up among patients who had received bezlotoxumab in MODIFY 2 and entered an extension substudy. Bezlotoxumab has low immunogenicity and is generally well tolerated, although the potential for heart failure in some patients requires consideration; cost-effectiveness data for bezlotoxumab are awaited with interest. Thus, a single intravenous infusion of bezlotoxumab during SoC antibacterial treatment for CDI is an emerging option for reducing CDI recurrence in adults at high risk of recurrence.

  9. Effects of Clostridium difficile infection in patients with alcoholic hepatitis.

    Science.gov (United States)

    Sundaram, Vinay; May, Folasade P; Manne, Vignan; Saab, Sammy

    2014-10-01

    Infection increases mortality in patients with alcoholic hepatitis (AH). Little is known about the association between Clostridium difficile infection (CDI) and AH. We examined the prevalence and effects of CDI in patients with AH, compared with those of other infections. We performed a cross-sectional analysis using data collected from the Nationwide Inpatient Sample, from 2008 through 2011. International Classification of Diseases, 9th revision, Clinical Modification codes were used to identify patients with AH. We used multivariable logistic regression to determine risk factors that affect mortality, negative binomial regression to evaluate the effects of CDI on predicted length of stay (LOS), and Poisson regression to determine the effects of CDI on predicted hospital charges. Chi-square and Wilcoxon rank-sum analyses were used to compare mortality, LOS, and hospital charges associated with CDI with those associated with urinary tract infection (UTI) and spontaneous bacterial peritonitis (SBP). Of 10,939 patients with AH, 177 had CDI (1.62%). Patients with AH and CDI had increased odds of inpatient mortality (adjusted odds ratio, 1.75; P = .04), a longer predicted LOS (10.63 vs 5.75 d; P effects appear similar to those for UTI and SBP. We propose further studies to determine the cost effectiveness of screening for CDI among patients with AH. Copyright © 2014 AGA Institute. Published by Elsevier Inc. All rights reserved.

  10. Fidaxomicin in Clostridium difficile infection: latest evidence and clinical guidance.

    Science.gov (United States)

    Mullane, Kathleen

    2014-03-01

    The incidence of Clostridium difficile infection (CDI) has risen 400% in the last decade. It currently ranks as the third most common nosocomial infection. CDI has now crossed over as a community-acquired infection. The major failing of current therapeutic options for the management of CDI is recurrence of disease after the completion of treatment. Fidaxomicin has been proven to be superior to vancomycin in successful sustained clinical response to therapy. Improved outcomes may be due to reduced collateral damage to the gut microflora by fidaxomicin, bactericidal activity, inhibition of Clostridial toxin formation and inhibition of new sporulation. This superiority is maintained in groups previously reported as being at high risk for CDI recurrence including those: with relapsed infection after a single treatment course; on concomitant antibiotic therapy; aged >65 years; with cancer; and with chronic renal insufficiency. Because the acquisition cost of fidaxomicin far exceeds that of metronidazole or vancomycin, in order to rationally utilize this agent, it should be targeted to those populations who are at high risk for relapse and in whom the drug has demonstrated superiority. In this manuscript is reviewed the changing epidemiology of CDI, current treatment options for this infection, proposed benefits of fidaxomicin over currently available antimicrobial options, available analysis of cost effectiveness of the drug, and is given recommendations for judicious use of the drug based upon the available published literature.

  11. Cost-effectiveness in Clostridium difficile treatment decision-making.

    Science.gov (United States)

    Nuijten, Mark Jc; Keller, Josbert J; Visser, Caroline E; Redekop, Ken; Claassen, Eric; Speelman, Peter; Pronk, Marja H

    2015-11-16

    To develop a framework for the clinical and health economic assessment for management of Clostridium difficile infection (CDI). CDI has vast economic consequences emphasizing the need for innovative and cost effective solutions, which were aim of this study. A guidance model was developed for coverage decisions and guideline development in CDI. The model included pharmacotherapy with oral metronidazole or oral vancomycin, which is the mainstay for pharmacological treatment of CDI and is recommended by most treatment guidelines. A design for a patient-based cost-effectiveness model was developed, which can be used to estimate the cost-effectiveness of current and future treatment strategies in CDI. Patient-based outcomes were extrapolated to the population by including factors like, e.g., person-to-person transmission, isolation precautions and closing and cleaning wards of hospitals. The proposed framework for a population-based CDI model may be used for clinical and health economic assessments of CDI guidelines and coverage decisions for emerging treatments for CDI.

  12. Cellular Uptake of the Clostridium perfringens Binary Iota-Toxin

    Science.gov (United States)

    Blöcker, Dagmar; Behlke, Joachim; Aktories, Klaus; Barth, Holger

    2001-01-01

    The binary iota-toxin is produced by Clostridium perfringens type E strains and consists of two separate proteins, the binding component iota b (98 kDa) and an actin-ADP-ribosylating enzyme component iota a (47 kDa). Iota b binds to the cell surface receptor and mediates the translocation of iota a into the cytosol. Here we studied the cellular uptake of iota-toxin into Vero cells. Bafilomycin A1, but not brefeldin A or nocodazole, inhibited the cytotoxic effects of iota-toxin, indicating that toxin is translocated from an endosomal compartment into the cytoplasm. Acidification (pH ≤ 5.0) of the extracellular medium enabled iota a to directly enter the cytosol in the presence of iota b. Activation by chymotrypsin induced oligomerization of iota b in solution. An average mass of 530 ± 28 kDa for oligomers was determined by analytical ultracentrifugation, indicating heptamer formation. The entry of iota-toxin into polarized CaCo-2 cells was studied by measuring the decrease in transepithelial resistance after toxin treatment. Iota-toxin led to a significant decrease in resistance when it was applied to the basolateral surface of the cells but not following application to the apical surface, indicating a polarized localization of the iota-toxin receptor. PMID:11292715

  13. Detection of toxigenic Clostridium difficile in paediatric patients.

    Science.gov (United States)

    Falces-Romero, Iker; Troyano-Hernáez, Paloma; García-Bujalance, Silvia; Baquero-Artigao, Fernando; Mellado-Peña, María José; García-Rodríguez, Julio

    2017-07-06

    Our main objective was a revision of clinical, microbiological and epidemiological results of Clostridium difficile-associated infection in paediatric patients (2010-2015). We compared the diagnoses performed by detection of toxins in feces and those performed by real-time PCR. This retrospective study included 82 paediatric patients. Detection of toxigenic C. difficile was performed sequentially, in diarrheal feces and under clinical request. A total of 39% of the patients were attended at Haematology-oncology Unit and >50% of them had previously received cephalosporins. Fever associated with diarrhea was more frequent in the group of toxin detection, whereas not receiving specific antibiotic treatment was more frequent in the group of positive PCR, without statistically significant differences. We highlight the presence of C. difficile infection in children under 2years old. A diagnostic testing in selected paediatric patients would be advisable when there is clinical suspicion of infection. Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  14. Spore Coat Architecture of Clostridium novyi-NT spores

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; McCafferey, J; Cheong, I; Huang, X; Bettegowda, C; Kinzler, K; Zhou, S; Vogelstein, B; Malkin, A

    2007-05-07

    Spores of the anaerobic bacterium Clostridium novyi-NT are able to germinate in and destroy hypoxic regions of tumors in experimental animals. Future progress in this area will benefit from a better understanding of the germination and outgrowth processes that are essential for the tumorilytic properties of these spores. Towards this end, we have used both transmission electron microscopy and atomic force microscopy to determine the structure of dormant as well as germinating spores. We found that the spores are surrounded by an amorphous layer intertwined with honeycomb parasporal layers. Moreover, the spore coat layers had apparently self-assembled and this assembly was likely to be governed by crystal growth principles. During germination and outgrowth, the honeycomb layers as well as the underlying spore coat and undercoat layers sequentially dissolved until the vegetative cell was released. In addition to their implications for understanding the biology of C. novyi-NT, these studies document the presence of proteinaceous growth spirals in a biological organism.

  15. Learning from Clostridium novyi-NT: How to defeat cancer

    Directory of Open Access Journals (Sweden)

    Li Wang

    2018-01-01

    Full Text Available Side effects associated with conventional anticancer therapies have prompted the new idea of solid tumor treatment strategy. One of them is using bacteria explored as potential antitumor agents over more than one century. Notably, the ideal therapy is a specifical target to tumors with limited toxicity. Here, we take “Clostridium novyi” for the search keyword in the PubMed from 2000 to 2015 and describe that C. novyi-NT spores act as “Trojan horse” for bacteriolytic therapy. This therapy is based on the fact that the live and attenuated obligate anaerobic bacteria are capable of binary fission selectively in anoxic areas of solid tumors and direct tumoricidal effects. Our succinct review mainly concentrates on the potential mechanisms of combination bacteriolytic therapy, an effective and safe tumor therapy with the help of C. novyi-NT. Importantly, C. novyi-NT spores were shown to induce solid tumor regression and exhibit the property to initiate an immune response. Therefore, C. novyi-NT spores should be an effective and safe tumor therapy.

  16. The Regulatory Networks That Control Clostridium difficile Toxin Synthesis

    Science.gov (United States)

    Martin-Verstraete, Isabelle; Peltier, Johann; Dupuy, Bruno

    2016-01-01

    The pathogenic clostridia cause many human and animal diseases, which typically arise as a consequence of the production of potent exotoxins. Among the enterotoxic clostridia, Clostridium difficile is the main causative agent of nosocomial intestinal infections in adults with a compromised gut microbiota caused by antibiotic treatment. The symptoms of C. difficile infection are essentially caused by the production of two exotoxins: TcdA and TcdB. Moreover, for severe forms of disease, the spectrum of diseases caused by C. difficile has also been correlated to the levels of toxins that are produced during host infection. This observation strengthened the idea that the regulation of toxin synthesis is an important part of C. difficile pathogenesis. This review summarizes our current knowledge about the regulators and sigma factors that have been reported to control toxin gene expression in response to several environmental signals and stresses, including the availability of certain carbon sources and amino acids, or to signaling molecules, such as the autoinducing peptides of quorum sensing systems. The overlapping regulation of key metabolic pathways and toxin synthesis strongly suggests that toxin production is a complex response that is triggered by bacteria in response to particular states of nutrient availability during infection. PMID:27187475

  17. Fidaxomicin - the new drug for Clostridium difficile infection

    Directory of Open Access Journals (Sweden)

    Chetana Vaishnavi

    2015-01-01

    Full Text Available Clostridium difficile is one of the many aetiological agents of antibiotic associated diarrhoea and is implicated in 15-25 per cent of the cases. The organism is also involved in the exacearbation of inflammatory bowel disease and extracolonic manifestations. Due to increase in the incidence of C. difficile infection (CDI, emergence of hypervirulent strains, and increased frequency of recurrence, the clinical management of the disease has become important. The management of CDI is based on disease severity, and current antibiotic treatment options are limited to vancomycin or metronidazole in the developing countries. this review article briefly describes important aspects of CDI, and the new drug, fidaxomicin, for its treatment. Fidaxomicin is particularly active against C.difficile and acts by inhibition of RNA synthesis. Clinical trials done to compare the efficacy and safety of fidaxomicin with that of vancomycin in treating CDI concluded that fidaxomicin was non-inferior to vancomycin for treatment of CDI and that there was a significant reduction in recurrences. The bactericidal properties of fidaxomicin make it an ideal alternative for CDI treatment. However, fidaxomicin use should be considered taking into account the potential benefits of the drug, along with the medical requirements of the patient, the risks of treatment and the high cost of fidaxomicin compared to other treatment regimens.

  18. [Nosocomial Clostridium difficile diarrhea--adverse effect of antibiotic therapy].

    Science.gov (United States)

    Lemeni, Daniela

    2010-01-01

    C. difficile is recognised as the main cause for colitis in hospitalised patients which are treated with antibiotics, chemotherapics or other drugs that disturb intestinal microbiota. Thus, a rapid and correct diagnostic of Clostridium difficile infections is essential for preventing nosocomial infection spread. Empiric therapy, regardless of the laboratory investigation results, is inadequate, especially in epidemic situations, as not all the cases of diarrhoea are due to C. difficile infection. Other risk factors for CDAD (Clostridiumn difficile Associated Diseases might be: prolonged hospitalization or residency in an asylum, age, existence of a severe chronic disease in the background nasogastric intubation, anti-ulcer drugs, at less extent gastrointestinal surgery, other immunosuppresive compounds etc. In our country, C. difficile infection is rather frequent in adults, though it is not always reported by clinicians. The circulation of endemic rybotype 027 in Romania is not well documented, the rybotype being extremely virulent and spread in other European countries. Hence the importance of extending the diagnostic capacity of C. difficile infection in order to allow detection of this rybotype among the strains isolated in our country.

  19. Investigation of inactivation of Clostridium botulinum toxin by nuclear radiation

    International Nuclear Information System (INIS)

    Kaltenhaeuser, A.; Werner, K.H.

    1989-01-01

    The effect of nuclear radiation on the toxicity and the molecular structure of the toxin produced by the microorganism Clostridium botulinum type A was investigated. The radiation induced changes in the structure of the toxin molecule. This effect is influenced by the composition or the medium above the toxin solution as well as by the temperature during the irradiation. The results of the investigation indicate that with increasing irradiation dose a new molecule was formed with immunological properties similar to the properties of the original molecule however with a greater molecular weight. After exposure to a radiation dose of 3,4 Mrad at normal temperature in air, complete detoxification of the substance was found. Immunizing experiments with the toxoid with two guinea-pigs indicated a pronounced increase of the antibody titer in the serum after 4 weeks. Vaccination experiments with the toxoid on animals show, that the protection against the effect of the toxin corresponds to the demands of the European Pharmacopoeia. The efficiency of the toxoid shows a similar efficiency as toxoids produced by chemical methods. The production of a toxoid-viccine with the relatively simple method of nuclear radiation appears possible. (orig./MG) With 12 refs., 3 tabs., 11 figs [de

  20. Prevention of Clostridium difficile infection in rural hospitals.

    Science.gov (United States)

    Haun, Nicholas; Hofer, Adam; Greene, M Todd; Borlaug, Gwen; Pritchett, Jenny; Scallon, Tina; Safdar, Nasia

    2014-03-01

    Prevention of Clostridium difficile infection (CDI) remains challenging across the spectrum of health care. There are limited data on prevention practices for CDI in the rural health care setting. An electronic survey was administered to 21 rural facilities in Wisconsin, part of the Rural Wisconsin Health Cooperative. Data were collected on hospital characteristics and practices to prevent endemic CDI. Fifteen facilities responded (71%). Nearly all respondent facilities reported regular use of dedicated patient care items, use of gown and gloves, private patient rooms, hand hygiene, and room cleaning. Facilities in which the infection preventionist thought the support of his/her leadership to be "Very good" or "Excellent" employed significantly more CDI practices (13.3 ± 2.4 [standard deviation]) compared with infection preventionists who thought there was less support from leadership (9.8 ± 3.0, P = .033). Surveillance for CDI was highly variable. The most frequent barriers to implementation of CDI prevention practices included lack of adequate resources, lack of a physician champion, and difficulty keeping up with new recommendations. Although most rural facilities in our survey reported using evidence-based practices for prevention of CDI, surveillance practices were highly variable, and data regarding the impact of these practices on CDI rates were limited. Future efforts that correlate CDI prevention initiatives and CDI incidence will help develop evidence-based practices in these resource-limited settings. Published by Mosby, Inc.

  1. Clostridium Difficile Infection Due to Pneumonia Treatment: Mortality Risk Models.

    Science.gov (United States)

    Chmielewska, M; Zycinska, K; Lenartowicz, B; Hadzik-Błaszczyk, M; Cieplak, M; Kur, Z; Wardyn, K A

    2017-01-01

    One of the most common gastrointestinal infection after the antibiotic treatment of community or nosocomial pneumonia is caused by the anaerobic spore Clostridium difficile (C. difficile). The aim of this study was to retrospectively assess mortality due to C. difficile infection (CDI) in patients treated for pneumonia. We identified 94 cases of post-pneumonia CDI out of the 217 patients with CDI. The mortality issue was addressed by creating a mortality risk models using logistic regression and multivariate fractional polynomial analysis. The patients' demographics, clinical features, and laboratory results were taken into consideration. To estimate the influence of the preceding respiratory infection, a pneumonia severity scale was included in the analysis. The analysis showed two statistically significant and clinically relevant mortality models. The model with the highest prognostic strength entailed age, leukocyte count, serum creatinine and urea concentration, hematocrit, coexisting neoplasia or chronic obstructive pulmonary disease. In conclusion, we report on two prognostic models, based on clinically relevant factors, which can be of help in predicting mortality risk in C. difficile infection, secondary to the antibiotic treatment of pneumonia. These models could be useful in preventive tailoring of individual therapy.

  2. Quantification of Nonproteolytic Clostridium botulinum Spore Loads in Food Materials.

    Science.gov (United States)

    Barker, Gary C; Malakar, Pradeep K; Plowman, June; Peck, Michael W

    2016-01-04

    We have produced data and developed analysis to build representations for the concentration of spores of nonproteolytic Clostridium botulinum in materials that are used during the manufacture of minimally processed chilled foods in the United Kingdom. Food materials are categorized into homogenous groups which include meat, fish, shellfish, cereals, fresh plant material, dairy liquid, dairy nonliquid, mushroom and fungi, and dried herbs and spices. Models are constructed in a Bayesian framework and represent a combination of information from a literature survey of spore loads from positive-control experiments that establish a detection limit and from dedicated microbiological tests for real food materials. The detection of nonproteolytic C. botulinum employed an optimized protocol that combines selective enrichment culture with multiplex PCR, and the majority of tests on food materials were negative. Posterior beliefs about spore loads center on a concentration range of 1 to 10 spores kg(-1). Posterior beliefs for larger spore loads were most significant for dried herbs and spices and were most sensitive to the detailed results from control experiments. Probability distributions for spore loads are represented in a convenient form that can be used for numerical analysis and risk assessments. Copyright © 2016 Barker et al.

  3. Probiotics for the treatment of Clostridium difficile associated disease

    Science.gov (United States)

    Fitzpatrick, Leo R

    2013-01-01

    The purpose of this review paper is to update the current and potential future role of probiotics for Clostridium difficile-associated disease (CDAD). Included in this review, is an update on the testing of newer probiotics (e.g., Bacillus coagulans GBI-30, 6086) in animal models of CDAD. There is a focus on the modulation of signal transduction pathways (i.e., transcription factors like cAMP response element-binding, activator protein 1, and nuclear factor kappa B), as well as the inhibition of certain kinases (e.g., p38 mitogen activated protein kinases) by probiotics. Inhibition of signal transduction by probiotics, such as Saccharomyces boulardii, result in multiple effects on intestinal fluid secretion, neutrophil influx into the colon, inflammation, and colonocyte apoptosis that may positively impact CDAD. Recent clinical approaches with probiotics, for the prevention of primary and recurrent CDAD, are also summarized in this review paper. Future directions for the treatment of CDAD by probiotics are also mentioned in this review. In particular, the use of multi-strain probiotic formulations such as Ecologic® AAD and VSL #3® may represent a rationale pharmacological approach, particularly as adjunctive therapies for CDAD. Understanding the mechanistic basis of CDAD, and how probiotics interfere at ceratin steps in the pathogenic process, may also present the opportunity to design other multi-strain probiotics that could have a future impact on CDAD. PMID:23946887

  4. Investigation of Clostridium botulinum group III's mobilome content.

    Science.gov (United States)

    Woudstra, Cédric; Le Maréchal, Caroline; Souillard, Rozenn; Anniballi, Fabrizio; Auricchio, Bruna; Bano, Luca; Bayon-Auboyer, Marie-Hélène; Koene, Miriam; Mermoud, Isabelle; Brito, Roseane B; Lobato, Francisco C F; Silva, Rodrigo O S; Dorner, Martin B; Fach, Patrick

    2018-02-01

    Clostridium botulinum group III is mainly responsible for botulism in animals. It could lead to high animal mortality rates and, therefore, represents a major environmental and economic concern. Strains of this group harbor the botulinum toxin locus on an unstable bacteriophage. Since the release of the first complete C. botulinum group III genome sequence (strain BKT015925), strains have been found to contain others mobile elements encoding for toxin components. In this study, seven assays targeting toxin genes present on the genetic mobile elements of C. botulinum group III were developed with the objective to better characterize C. botulinum group III strains. The investigation of 110 C. botulinum group III strains and 519 naturally contaminated samples collected during botulism outbreaks in Europe showed alpha-toxin and C2-I/C2-II markers to be systematically associated with type C/D bont-positive samples, which may indicate an important role of these elements in the pathogenicity mechanisms. On the contrary, bont type D/C strains and the related positive samples appeared to contain almost none of the markers tested. Interestingly, 31 bont-negative samples collected on farms after a botulism outbreak revealed to be positive for some of the genetic mobile elements tested. This suggests loss of the bont phage, either in farm environment after the outbreak or during laboratory handling. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. [Current treatment and epidemiology of Clostridium difficile infections].

    Science.gov (United States)

    Dinh, A; Bouchand, F; Le Monnier, A

    2015-09-01

    During the past 10years, Clostridium difficile infections (CDI) have become a major public health challenge. Their epidemiology has changed with a rise in the number of cases and an increase in severe episodes. Recurrence and failure of conventional treatments have become more common. Furthermore, a spread of CDI has been observed in the general population-involving subjects without the usual risk factors (unexposed to antibiotic treatment, young people, pregnant women, etc.). All these change are partially due to the emergence of the hypervirulent and hyperepidemic clone NAP1/B1/027. New therapeutic strategies (antimicrobial treatment, immunoglobulins, toxin chelation, fecal microbiota transplantation) are now available and conventional treatments (metronidazole and vancomycin) have been reevaluated with new recommendations. Recent studies show a better efficacy of vancomycin compared to metronidazole for severe episodes. Fidaxomicin is a novel antibiotic drug with interesting features, including an efficacy not inferior to vancomycin and a lower risk of recurrence. Finally, for multi-recurrent forms, fecal microbiota transplantation seems to be the best option. We present the available data in this review. Copyright © 2015 Société nationale française de médecine interne (SNFMI). Published by Elsevier SAS. All rights reserved.

  6. DOMESTIC BUTANOL-PRODUCING STRAINS OF THE Clostridium GENUS

    Directory of Open Access Journals (Sweden)

    O. O. Tigunova

    2017-02-01

    Full Text Available The aim of the work was to summarize the results of own research concerning obtaining butanol producing strains of Clostridium genus, to identify them by physiological, morphological and genetic methods. Further study of characteristics and biological features of the strains, and various approaches in biotechnological process of butanol production are discussed. The work includes methods to increase butanol accumulation by producer strains. Perspectives of using chemical mutagenesis in Clostridia as a method of increasing butanol production are considered. The feasibility of using non-food raw material as a substrate for fermentation is discussed. Different methods of pretreatment and their impact on the accumulation of butanol in the liquid medium are compared. Butanol accumulation is shown to increase significantly if the synthesis precursors are added as components of enzymatic medium, and the “reverse bard” is used to reduce waste production without affecting the level of butanol synthesis. The problem of conservation of producing strains is given, and protective medium for microorganisms during the freeze-drying is defined.

  7. Pancreatitis caused by Clostridium perfringens infection with extensive retropneumoperitoneum

    International Nuclear Information System (INIS)

    Merchante, E.; Garcia, F. J.; Perez, H.; Marquez, J. L.

    2001-01-01

    We present a case of primary emphysematous pancreatitis caused by Clostridium perfringens infection (also Known as spontaneous pancreatic gas gangrene) in a 66-year-old man with diabetes and a history of recurrent pancreatitis. One notable feature is the absence of a focal distribution, which is seen on radiological studies to be accompanied by extensive retropneumoperitoneum, with dissemination of the gas toward the mesenteric root and pelvic extra peritoneal spaces. This wide diffusion is aided by the C. perfringens toxins and the pancreatic enzymes released, leading to a fulminate course, an elevated rate of early mortality among the cases reviewed. The early diagnosis of this disease is fundamental, enabling aggressive medical treatment and emergency surgery. Diabetes is a known risk factor for anaerobic infection, including C. perfringens, as in the case of emphysematous cholecystitis. A diseased pancreas or pancreatic duct facilitates the development of infections since it eliminates poorly the microorganisms that reach it from the duodenum. Gas gangrene secondary to necrosis-related super infection or pancreatic collections is uncommon, and spontaneous or primary cases are exceptionally are. (Author) 13 refs

  8. Clostridium difficile in retail meat and processing plants in Texas.

    Science.gov (United States)

    Harvey, Roger B; Norman, Keri N; Andrews, Kathleen; Norby, Bo; Hume, Michael E; Scanlan, Charles M; Hardin, Margaret D; Scott, Harvey M

    2011-07-01

    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 to human beings. The objective of the present study was to determine the prevalence of C. difficile in pork from sausage manufacturing plants and retail meat in Texas. Twenty-three C. difficile isolates were detected from 243 meat samples (9.5%) from 3 sausage-manufacturing plants and 5 retail meat outlets from 2004 to 2009. Twenty-two isolates were positive for toxins A, B, and binary toxin, and were characterized as toxinotype V, PFGE type-NAP7, or "NAP7-variant." Susceptibilities to 11 antimicrobial agents in the current study were similar to those reported previously for toxinotype V isolates, although the results suggested somewhat reduced resistance than reported for other meat, animal, or human clinical toxinotype V isolates.

  9. Experimental Clostridium perfringens type D enterotoxemia in goats.

    Science.gov (United States)

    Uzal, F A; Kelly, W R

    1998-03-01

    The effects of intraduodenal administration of Clostridium perfringens cultures and culture products in goats were evaluated to develop a reliable experimental model of enterotoxemia in this species. Five conventionally reared, 11-16-week-old Angora goat kids were dosed intraduodenally with whole cultures of C. perfringens type D; five similar animals were dosed with C. perfringens type D filtered culture supernatant; and a third group of five kids was dosed with C. perfringens type D washed cells. Two kids were used as controls and received sterile, nontoxic culture medium intraduodenally. All animals received starch solution into the abomasum. All five kids inoculated with whole culture and three of five dosed with culture supernatant and with washed cells developed central nervous system signs. Diarrhea was observed in two of five kids inoculated with whole culture, in all five of those dosed with culture supernatant, and in three of five of those that received washed cells. The most striking postmortem findings consisted of lung edema, necrotizing pseudomembranous colitis, and cerebral vasogenic edema. The protocol thus provided a reasonable model of naturally occurring enterotoxemia in goats, producing a range of clinical signs and postmortem changes similar to those observed in the natural disease.

  10. Muricholic acids inhibit Clostridium difficile spore germination and growth.

    Directory of Open Access Journals (Sweden)

    Michael B Francis

    Full Text Available Infections caused by Clostridium difficile have increased steadily over the past several years. While studies on C. difficile virulence and physiology have been hindered, in the past, by lack of genetic approaches and suitable animal models, newly developed technologies and animal models allow these processes to be studied in detail. One such advance is the generation of a mouse-model of C. difficile infection. The development of this system is a major step forward in analyzing the genetic requirements for colonization and infection. While important, it is equally as important in understanding what differences exist between mice and humans. One of these differences is the natural bile acid composition. Bile acid-mediated spore germination is an important step in C. difficile colonization. Mice produce several different bile acids that are not found in humans. These muricholic acids have the potential to impact C. difficile spore germination. Here we find that the three muricholic acids (α-muricholic acid, β-muricholic acid and ω-muricholic acid inhibit C. difficile spore germination and can impact the growth of vegetative cells. These results highlight an important difference between humans and mice and may have an impact on C. difficile virulence in the mouse-model of C. difficile infection.

  11. Clostridium difficile in Crete, Greece: epidemiology, microbiology and clinical disease.

    Science.gov (United States)

    Samonis, G; Vardakas, K Z; Tansarli, G S; Dimopoulou, D; Papadimitriou, G; Kofteridis, D P; Maraki, S; Karanika, M; Falagas, M E

    2016-01-01

    We studied the epidemiology and microbiology of Clostridium difficile and the characteristics of patients with C. difficile infection (CDI) in Crete in three groups of hospitalized patients with diarrhoea: group 1 [positive culture and positive toxin by enzyme immunoassay (EIA)]; group 2 (positive culture, negative toxin); group 3 (negative culture, negative toxin). Patients in group 1 were designated as those with definitive CDI (20 patients for whom data was available) and matched with cases in group 2 (40 patients) and group 3 (40 patients). C. difficile grew from 6% (263/4379) of stool specimens; 14·4% of these had positive EIA, of which 3% were resistant to metronidazole. Three isolates had decreased vancomycin susceptibility. Patients in groups 1 and 2 received more antibiotics (P = 0·03) and had more infectious episodes (P = 0·03) than patients in group 3 prior to diarrhoea. Antibiotic administration for C. difficile did not differ between groups 1 and 2. Mortality was similar in all three groups (10%, 12·5% and 5%, P = 0·49). CDI frequency was low in the University Hospital of Crete and isolates were susceptible to metronidazole and vancomycin.

  12. Phosphoproteomic investigation of a solvent producing bacterium Clostridium acetobutylicum.

    Science.gov (United States)

    Bai, Xue; Ji, Zhihong

    2012-07-01

    In this study, we employed TiO₂ enrichment and high accuracy liquid chromatography-mass spectrometry-mass spectrometry to identify the phosphoproteome of Clostridium acetobutyicum ATCC824 in acidogenesis and solventogenesis. As many as 82 phosphopeptides in 61 proteins, with 107 phosphorylated sites on serine, threonine, or tyrosine, were identified with high confidence. We detected 52 phosphopeptides from 44 proteins in acidogenesis and 70 phosphopeptides from 51 proteins in solventogenesis, respectively. Bioinformatic analysis revealed most of the phosphoproteins located in cytoplasm and participated in carbon metabolism. Based on comparison between the two stages, we found 27 stage-specific phosphorylated proteins (10 in acidogenesis and 17 in solventogenesis), some of which were solvent production-related enzymes and metabolic regulators, showed significantly different phosphorylated status. Further analysis indicated that protein phosphorylation could be involved in the shift of stages or in solvent production pathway directly. Comparison against several other organisms revealed the evolutionary diversity among them on phosphorylation level in spite of their high homology on protein sequence level.

  13. Clostridium difficile infection: epidemiology, diagnosis and understanding transmission.

    Science.gov (United States)

    Martin, Jessica S H; Monaghan, Tanya M; Wilcox, Mark H

    2016-04-01

    Clostridium difficile infection (CDI) continues to affect patients in hospitals and communities worldwide. The spectrum of clinical disease ranges from mild diarrhoea to toxic megacolon, colonic perforation and death. However, this bacterium might also be carried asymptomatically in the gut, potentially leading to 'silent' onward transmission. Modern technologies, such as whole-genome sequencing and multi-locus variable-number tandem-repeat analysis, are helping to track C. difficile transmission across health-care facilities, countries and continents, offering the potential to illuminate previously under-recognized sources of infection. These typing strategies have also demonstrated heterogeneity in terms of CDI incidence and strain types reflecting different stages of epidemic spread. However, comparison of CDI epidemiology, particularly between countries, is challenging due to wide-ranging approaches to sampling and testing. Diagnostic strategies for C. difficile are complicated both by the wide range of bacterial targets and tests available and the need to differentiate between toxin-producing and non-toxigenic strains. Multistep diagnostic algorithms have been recommended to improve sensitivity and specificity. In this Review, we describe the latest advances in the understanding of C. difficile epidemiology, transmission and diagnosis, and discuss the effect of these developments on the clinical management of CDI.

  14. The potential for emerging therapeutic options for Clostridium difficile infection

    Science.gov (United States)

    Mathur, Harsh; Rea, Mary C; Cotter, Paul D; Ross, R Paul; Hill, Colin

    2014-01-01

    Clostridium difficile is mainly a nosocomial pathogen and is a significant cause of antibiotic-associated diarrhea. It is also implicated in the majority of cases of pseudomembranous colitis. Recently, advancements in next generation sequencing technology (NGS) have highlighted the extent of damage to the gut microbiota caused by broad-spectrum antibiotics, often resulting in C. difficile infection (CDI). Currently the treatment of choice for CDI involves the use of metronidazole and vancomycin. However, recurrence and relapse of CDI, even after rounds of metronidazole/vancomycin administration is a problem that must be addressed. The efficacy of alternative antibiotics such as fidaxomicin, rifaximin, nitazoxanide, ramoplanin and tigecycline, as well as faecal microbiota transplantation has been assessed and some have yielded positive outcomes against C. difficile. Some bacteriocins have also shown promising effects against C. difficile in recent years. In light of this, the potential for emerging treatment options and efficacy of anti-C. difficile vaccines are discussed in this review. PMID:25564777

  15. Current knowledge on the laboratory diagnosis of Clostridium difficile infection.

    Science.gov (United States)

    Martínez-Meléndez, Adrián; Camacho-Ortiz, Adrián; Morfin-Otero, Rayo; Maldonado-Garza, Héctor Jesús; Villarreal-Treviño, Licet; Garza-González, Elvira

    2017-03-07

    Clostridium difficile ( C. difficile ) is a spore-forming, toxin-producing, gram-positive anaerobic bacterium that is the principal etiologic agent of antibiotic-associated diarrhea. Infection with C. difficile (CDI) is characterized by diarrhea in clinical syndromes that vary from self-limited to mild or severe. Since its initial recognition as the causative agent of pseudomembranous colitis, C. difficile has spread around the world. CDI is one of the most common healthcare-associated infections and a significant cause of morbidity and mortality among older adult hospitalized patients. Due to extensive antibiotic usage, the number of CDIs has increased. Diagnosis of CDI is often difficult and has a substantial impact on the management of patients with the disease, mainly with regards to antibiotic management. The diagnosis of CDI is primarily based on the clinical signs and symptoms and is only confirmed by laboratory testing. Despite the high burden of CDI and the increasing interest in the disease, episodes of CDI are often misdiagnosed. The reasons for misdiagnosis are the lack of clinical suspicion or the use of inappropriate tests. The proper diagnosis of CDI reduces transmission, prevents inadequate or unnecessary treatments, and assures best antibiotic treatment. We review the options for the laboratory diagnosis of CDI within the settings of the most accepted guidelines for CDI diagnosis, treatment, and prevention of CDI.

  16. The host immune response to Clostridium difficile infection

    Science.gov (United States)

    2013-01-01

    Clostridium difficile infection (CDI) is the most common infectious cause of healthcare-acquired diarrhoea. Outcomes of C. difficile colonization are varied, from asymptomatic carriage to fulminant colitis and death, due in part to the interplay between the pathogenic virulence factors of the bacterium and the counteractive immune responses of the host. Secreted toxins A and B are the major virulence factors of C. difficile and induce a profound inflammatory response by intoxicating intestinal epithelial cells causing proinflammatory cytokine release. Host cell necrosis, vascular permeability and neutrophil infiltration lead to an elevated white cell count, profuse diarrhoea and in severe cases, dehydration, hypoalbuminaemia and toxic megacolon. Other bacterial virulence factors, including surface layer proteins and flagella proteins, are detected by host cell surface signal molecules that trigger downstream cell-mediated immune pathways. Human studies have identified a role for serum and faecal immunoglobulin levels in protection from disease, but the recent development of a mouse model of CDI has enabled studies into the precise molecular interactions that trigger the immune response during infection. Key effector molecules have been identified that can drive towards a protective anti-inflammatory response or a damaging proinflammatory response. The limitations of current antimicrobial therapies for CDI have led to the development of both active and passive immunotherapies, none of which have, as yet been formally approved for CDI. However, recent advances in our understanding of the molecular basis of host immune protection against CDI may provide an exciting opportunity for novel therapeutic developments in the future. PMID:25165542

  17. Survey of Clostridium difficile infection surveillance systems in Europe, 2011.

    Science.gov (United States)

    Kola, Axel; Wiuff, Camilla; Akerlund, Thomas; van Benthem, Birgit H; Coignard, Bruno; Lyytikäinen, Outi; Weitzel-Kage, Doris; Suetens, Carl; Wilcox, Mark H; Kuijper, Ed J; Gastmeier, Petra

    2016-07-21

    To develop a European surveillance protocol for Clostridium difficile infection (CDI), existing national CDI surveillance systems were assessed in 2011. A web-based electronic form was provided for all national coordinators of the European CDI Surveillance Network (ECDIS-Net). Of 35 national coordinators approached, 33 from 31 European countries replied. Surveillance of CDI was in place in 14 of the 31 countries, comprising 18 different nationwide systems. Three of 14 countries with CDI surveillance used public health notification of cases as the route of reporting, and in another three, reporting was limited to public health notification of cases of severe CDI. The CDI definitions published by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) and the European Centre for Disease Prevention and Control (ECDC) were widely used, but there were differing definitions to distinguish between community- and healthcare-associated cases. All CDI surveillance systems except one reported annual national CDI rates (calculated as number of cases per patient-days). Only four surveillance systems regularly integrated microbiological data (typing and susceptibility testing results). Surveillance methods varied considerably between countries, which emphasises the need for a harmonised European protocol to allow consistent monitoring of the CDI epidemiology at European level. The results of this survey were used to develop a harmonised EU-wide hospital-based CDI surveillance protocol. This article is copyright of The Authors, 2016.

  18. Management of inflammatory bowel disease with Clostridium difficile infection.

    Science.gov (United States)

    D'Aoust, Julie; Battat, Robert; Bessissow, Talat

    2017-07-21

    To address the management of Clostridium difficile ( C. difficile ) infection (CDI) in the setting of suspected inflammatory bowel disease (IBD)-flare. A systematic search of the Ovid MEDLINE and EMBASE databases by independent reviewers identified 70 articles including a total of 932141 IBD patients or IBD-related hospitalizations. In those with IBD, CDI is associated with increased morbidity, including subsequent escalation in IBD medical therapy, urgent colectomy and increased hospitalization, as well as excess mortality. Vancomycin-containing regimens are effective first-line therapies for CDI in IBD inpatients. No prospective data exists with regards to the safety or efficacy of initiating or maintaining corticosteroid, immunomodulator, or biologic therapy to treat IBD in the setting of CDI. Corticosteroid use is a risk factor for the development of CDI, while immunomodulators and biologics are not. Strong recommendations regarding when to initiate IBD specific therapy in those with CDI are precluded by a lack of evidence. However, based on expert opinion and observational data, initiation or resumption of immunosuppressive therapy after 48-72 h of targeted antibiotic treatment for CDI may be considered.

  19. The role of toxins in Clostridium difficile infection.

    Science.gov (United States)

    Chandrasekaran, Ramyavardhanee; Lacy, D Borden

    2017-11-01

    Clostridium difficile is a bacterial pathogen that is the leading cause of nosocomial antibiotic-associated diarrhea and pseudomembranous colitis worldwide. The incidence, severity, mortality and healthcare costs associated with C. difficile infection (CDI) are rising, making C. difficile a major threat to public health. Traditional treatments for CDI involve use of antibiotics such as metronidazole and vancomycin, but disease recurrence occurs in about 30% of patients, highlighting the need for new therapies. The pathogenesis of C. difficile is primarily mediated by the actions of two large clostridial glucosylating toxins, toxin A (TcdA) and toxin B (TcdB). Some strains produce a third toxin, the binary toxin C. difficile transferase, which can also contribute to C. difficile virulence and disease. These toxins act on the colonic epithelium and immune cells and induce a complex cascade of cellular events that result in fluid secretion, inflammation and tissue damage, which are the hallmark features of the disease. In this review, we summarize our current understanding of the structure and mechanism of action of the C. difficile toxins and their role in disease. Published by Oxford University Press on behalf of FEMS 2017.

  20. Control of Clostridium difficile infection by defined microbial communities

    Science.gov (United States)

    Collins, James

    2017-01-01

    Summary Each year in the United States, billions of dollars are spent combating almost half a million Clostridium difficile infections (CDI) and trying to reduce the ~29,000 patient deaths where C. difficile has an attributed role (1). In Europe, disease prevalence varies by country and level of surveillance, though yearly costs are estimated at €3 billion (2). One factor contributing to the significant healthcare burden of C. difficile is the relatively high frequency of recurrent C. difficile infections(3). Recurrent C. difficile infection (rCDI), i.e., a second episode of symptomatic CDI occurring within eight weeks of successful initial CDI treatment, occurs in ~25% of patients with 35-65% of these patients experiencing multiple episodes of recurrent disease(4, 5). Using microbial communities to treat rCDI, either as whole fecal transplants or as defined consortia of bacterial isolates have shown great success (in the case of fecal transplants) or potential promise (in the case of defined consortia of isolates). This review will briefly summarize the epidemiology and physiology of C. difficile infection, describe our current understanding of how fecal microbiota transplants treat recurrent CDI, and outline potential ways through which that knowledge can be used to rationally-design and test alternative microbe-based therapeutics. PMID:28936948

  1. Clostridium difficile colitis in patients undergoing lumbar spine surgery.

    Science.gov (United States)

    Skovrlj, Branko; Guzman, Javier Z; Silvestre, Jason; Al Maaieh, Motasem; Qureshi, Sheeraz A

    2014-09-01

    Retrospective database analysis. To investigate incidence, comorbidities, and impact on health care resources of Clostridium difficile infection after lumbar spine surgery. C. difficile colitis is reportedly increasing in hospitalized patients and can have a negative impact on patient outcomes. No data exist on estimates of C. difficile infection rates and its consequences on patient outcomes and health care resources among patients undergoing lumbar spine surgery. The Nationwide Inpatient Sample was examined from 2002 to 2011. Patients were included for study based on International Classification of Diseases, Ninth Revision, Clinical Modification, procedural codes for lumbar spine surgery for degenerative diagnoses. Baseline patient characteristics were determined and multivariable analyses assessed factors associated with increased incidence of C. difficile and risk of mortality. The incidence of C. difficile infection in patients undergoing lumbar spine surgery is 0.11%. At baseline, patients infected with C. difficile were significantly older (65.4 yr vs. 58.9 yr, Pinfection. Small hospital size was associated with decreased odds (odds ratio [OR], 0.5; Pinfection. Uninsured (OR, 1.62; Pinfection. C. difficile increased hospital length of stay by 8 days (Pdifficile infection after lumbar spine surgery carries a 36.4-fold increase in mortality and costs approximately $10,658,646 per year to manage. These data suggest that great care should be taken to avoid C. difficile colitis in patients undergoing lumbar spine surgery because it is associated with longer hospital stays, greater overall costs, and increased inpatient mortality. 3.

  2. A review of the economics of treating Clostridium difficile infection.

    Science.gov (United States)

    Mergenhagen, Kari A; Wojciechowski, Amy L; Paladino, Joseph A

    2014-07-01

    Clostridium difficile infection (CDI) is a costly result of antibiotic use, responsible for an estimated 14,000 deaths annually in the USA according to the Centers for Disease Control and Prevention. Annual costs attributable to CDI are in excess of $US 1 billion. This review summarizes appropriate utilization of prevention and treatment methods for CDI that have the potential to reduce the economic and humanistic costs of the disease. Some cost-effective strategies to prevent CDI include screening and isolation of hospital admissions based on C. difficile carriage to reduce transmission in the inpatient setting, and probiotics, which are potentially efficacious in preventing CDI in the appropriate patient population. The most extensively studied agents for treatment of CDI are metronidazole, vancomycin, and fidaxomicin. Most economic comparisons between metronidazole and vancomycin favor vancomycin, especially with the emergence of metronidazole-resistant C. difficile strains. Metronidazole can only be recommended for mild disease. Moderate to severe CDI should be treated with vancomycin, preferably the compounded oral solution, which provides the most cost-effective therapeutic option. Fidaxomicin offers a clinically effective and potentially cost-effective alternative for treating moderate CDI in patients who do not have the NAP1/BI/027 strain of C. difficile. Probiotics and fecal microbiota transplant have variable efficacy and the US FDA does not currently regulate the content; the potential economic advantages of these treatment modalities are currently unknown.

  3. Clostridium difficile infection in the elderly: an update on management.

    Science.gov (United States)

    Asempa, Tomefa E; Nicolau, David P

    2017-01-01

    The burden of Clostridium difficile infection (CDI) is profound and growing. CDI now represents a common cause of health care-associated diarrhea, and is associated with significant morbidity, mortality, and health care costs. CDI disproportionally affects the elderly, possibly explained by the following risk factors: age-related impairment of the immune system, increasing antibiotic utilization, and frequent health care exposure. In the USA, recent epidemiological studies estimate that two out of every three health care-associated CDIs occur in patients 65 years or older. Additionally, the elderly are at higher risk for recurrent CDI. Existing therapeutic options include metronidazole, oral vancomycin, and fidaxomicin. Choice of agent depends on disease severity, history of recurrence, and, increasingly, the drug cost. Bezlotoxumab, a recently approved monoclonal antibody targeting C. difficile toxin B, offers an exciting advancement into immunologic therapies. Similarly, fecal microbiota transplantation is gaining popularity as an effective option mainly for recurrent CDI. The challenge of decreasing CDI burden in the elderly involves adopting preventative strategies, optimizing initial treatment, and decreasing the risk of recurrence. Expanded strategies are certainly needed to improve outcomes in this high-risk population. This review considers available data from prospective and retrospective studies as well as case reports to illustrate the merits and gaps in care related to the management of CDI in the elderly.

  4. Clostridium difficile infection: current, forgotten and emerging treatment options.

    Science.gov (United States)

    Drekonja, Dimitri M

    2014-09-01

    Clostridium difficile infection (CDI) has increased in incidence and severity, and is now among the most common nosocomial infections. Several agents are available for the initial treatment of CDI, some of which are rarely used, and none of which is clearly superior for initial clinical cure. Fidaxomicin appears to offer a benefit in terms of preventing recurrent disease, although the cost-benefit ratio is debated. Recurrent CDI is a major challenge, occurring after 15-30% of initial episodes. The treatment of recurrent CDI is difficult, with sparse evidence available to support any particular agent. Fecal microbiota therapy, also known as 'stool transplantation', appears to be highly effective, although availability is currently limited, and the regulatory environment is in flux. Synthetic stool products and an orally available fecal microbiota therapy product are both under investigation, which may address the problem of availability. As with most infectious diseases, an effective vaccine would be a welcome addition to our armamentarium, but none is currently available.

  5. Prevalence and molecular epidemiology of Clostridium difficile infection in Indonesia

    Directory of Open Access Journals (Sweden)

    D.A. Collins

    2017-07-01

    Full Text Available Clostridium difficile has not been studied in detail in Asia, particularly Southeast Asia. We thus performed a prevalence study across four hospitals in Central Java province, Indonesia. Stool samples were collected from patients with diarrhoea and tested by enzyme immunoassay for glutamate dehydrogenase (GDH and toxin A/B (C DIFF QUIK CHEK COMPLETE, TechLab. Specimens were cultured and molecular typing was performed. In total, 340 samples were tested, of which 70 (20.6% were GDH positive, with toxin detected in 19 (5.6%. Toxigenic C. difficile was isolated from 37 specimens (10.9%, while a further 36 (10.6% nontoxigenic isolates were identified. The most common strain was ribotype 017 (24.3% of 74 isolates, followed by nontoxigenic types QX 224 (9.5%, and QX 238 and QX 108 (both 8.1%. The high prevalence of C. difficile highlights a need for ongoing surveillance of C. difficile infection in Indonesia.

  6. Sensitivity of clostridium acetobutylicum to oxygen and ionizing radiation

    International Nuclear Information System (INIS)

    Sozer, A.C.; Adler, H.I.; Machanoff, R.; Haney, S.

    1984-01-01

    The authors are studying the sensitivity of four strains of the obligate anaerobe, Clostridium acetobutylicum, to oxygen and ionizing radiation. Anaerobic bacteria are useful for such studies because of the absence of elaborate oxygen detoxification mechanisms that are found in aerobes. Their experiments make use of sterile membrane fragments from Escherichia coli that rapidly remove molecular oxygen from media and permit growth of anaerobes without the use of reducing agents or anaerobic chambers. Of the four strains examined for sensitivity to ionizing radiation under anaerobic conditions, one has an LD/sub 50/ of -- 25 krads and the others have an LD/sub 50/ of -- 7 krads. The radiation resistant strain is also relatively resistant to oxygen exposure. Sensitivity to oxygen was determined by diluting cells in buffer at 28 0 and bubbling with air. An exposure to air for 40 min induced only slight inactivation in the radiation resistant strain. All strains are capable of removing oxygen from complex media but there is no apparent correlation between this oxygen consuming reaction and inactivation by either oxygen or radiation

  7. The structure of the S-layer of Clostridium difficile.

    Science.gov (United States)

    Bradshaw, William J; Roberts, April K; Shone, Clifford C; Acharya, K Ravi

    2018-03-01

    The nosocomially acquired pathogen Clostridium difficile is the primary causative agent of antibiotic associated diarrhoea and causes tens of thousands of deaths globally each year. C. difficile presents a paracrystalline protein array on the surface of the cell known as an S-layer. S-layers have been demonstrated to possess a wide range of important functions, which, combined with their inherent accessibility, makes them a promising drug target. The unusually complex S-layer of C. difficile is primarily comprised of the high- and low- molecular weight S-layer proteins, HMW SLP and LMW SLP, formed from the cleavage of the S-layer precursor protein, SlpA, but may also contain up to 28 SlpA paralogues. A model of how the S-layer functions as a whole is required if it is to be exploited in fighting the bacterium. Here, we provide a summary of what is known about the S-layer of C. difficile and each of the paralogues and, considering some of the domains present, suggest potential roles for them.

  8. Metabolic engineering of Clostridium autoethanogenum for selective alcohol production.

    Science.gov (United States)

    Liew, Fungmin; Henstra, Anne M; Kӧpke, Michael; Winzer, Klaus; Simpson, Sean D; Minton, Nigel P

    2017-03-01

    Gas fermentation using acetogenic bacteria such as Clostridium autoethanogenum offers an attractive route for production of fuel ethanol from industrial waste gases. Acetate reduction to acetaldehyde and further to ethanol via an aldehyde: ferredoxin oxidoreductase (AOR) and alcohol dehydrogenase has been postulated alongside the classic pathway of ethanol formation via a bi-functional aldehyde/alcohol dehydrogenase (AdhE). Here we demonstrate that AOR is critical to ethanol formation in acetogens and inactivation of AdhE led to consistently enhanced autotrophic ethanol production (up to 180%). Using ClosTron and allelic exchange mutagenesis, which was demonstrated for the first time in an acetogen, we generated single mutants as well as double mutants for both aor and adhE isoforms to confirm the role of each gene. The aor1+2 double knockout strain lost the ability to convert exogenous acetate, propionate and butyrate into the corresponding alcohols, further highlighting the role of these enzymes in catalyzing the thermodynamically unfavourable reduction of carboxylic acids into alcohols. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  9. The economic impact of Clostridium difficile infection: a systematic review.

    Science.gov (United States)

    Nanwa, Natasha; Kendzerska, Tetyana; Krahn, Murray; Kwong, Jeffrey C; Daneman, Nick; Witteman, William; Mittmann, Nicole; Cadarette, Suzanne M; Rosella, Laura; Sander, Beate

    2015-04-01

    With Clostridium difficile infection (CDI) on the rise, knowledge of the current economic burden of CDI can inform decisions on interventions related to CDI. We systematically reviewed CDI cost-of-illness (COI) studies. We performed literature searches in six databases: MEDLINE, Embase, the Health Technology Assessment Database, the National Health Service Economic Evaluation Database, the Cost-Effectiveness Analysis Registry, and EconLit. We also searched gray literature and conducted reference list searches. Two reviewers screened articles independently. One reviewer abstracted data and assessed quality using a modified guideline for economic evaluations. The second reviewer validated the abstraction and assessment. We identified 45 COI studies between 1988 and June 2014. Most (84%) of the studies were from the United States, calculating costs of hospital stays (87%), and focusing on direct costs (100%). Attributable mean CDI costs ranged from $8,911 to $30,049 for hospitalized patients. Few studies stated resource quantification methods (0%), an epidemiological approach (0%), or a justified study perspective (16%) in their cost analyses. In addition, few studies conducted sensitivity analyses (7%). Forty-five COI studies quantified and confirmed the economic impact of CDI. Costing methods across studies were heterogeneous. Future studies should follow standard COI methodology, expand study perspectives (e.g., patient), and explore populations least studied (e.g., community-acquired CDI).

  10. Solution preparation

    International Nuclear Information System (INIS)

    Seitz, M.G.

    1982-01-01

    Reviewed in this statement are methods of preparing solutions to be used in laboratory experiments to examine technical issues related to the safe disposal of nuclear waste from power generation. Each approach currently used to prepare solutions has advantages and any one approach may be preferred over the others in particular situations, depending upon the goals of the experimental program. These advantages are highlighted herein for three approaches to solution preparation that are currently used most in studies of nuclear waste disposal. Discussion of the disadvantages of each approach is presented to help a user select a preparation method for his particular studies. Also presented in this statement are general observations regarding solution preparation. These observations are used as examples of the types of concerns that need to be addressed regarding solution preparation. As shown by these examples, prior to experimentation or chemical analyses, laboratory techniques based on scientific knowledge of solutions can be applied to solutions, often resulting in great improvement in the usefulness of results

  11. Discovery of a novel gene involved in autolysis of Clostridium cells.

    Science.gov (United States)

    Yang, Liejian; Bao, Guanhui; Zhu, Yan; Dong, Hongjun; Zhang, Yanping; Li, Yin

    2013-06-01

    Cell autolysis plays important physiological roles in the life cycle of clostridial cells. Understanding the genetic basis of the autolysis phenomenon of pathogenic Clostridium or solvent producing Clostridium cells might provide new insights into this important species. Genes that might be involved in autolysis of Clostridium acetobutylicum, a model clostridial species, were investigated in this study. Twelve putative autolysin genes were predicted in C. acetobutylicum DSM 1731 genome through bioinformatics analysis. Of these 12 genes, gene SMB_G3117 was selected for testing the in tracellular autolysin activity, growth profile, viable cell numbers, and cellular morphology. We found that overexpression of SMB_G3117 gene led to earlier ceased growth, significantly increased number of dead cells, and clear electrolucent cavities, while disruption of SMB_G3117 gene exhibited remarkably reduced intracellular autolysin activity. These results indicate that SMB_G3117 is a novel gene involved in cellular autolysis of C. acetobutylicum.

  12. Analysis of Clostridium difficile infections in patients hospitalized at the nephrological ward in Poland

    Directory of Open Access Journals (Sweden)

    Agata Kujawa-Szewieczek

    2016-05-01

    Full Text Available Background: Few studies have evaluated the incidence and risk factors of Clostridium difficile infection (CDI in the adult Polish population, in particular in solid organ recipients hospitalized at the nephrological ward.Aim: The aim of this study was to analyze Clostridium difficile infections (CDI among patients hospitalized in the Department of Nephrology, Transplantation and Internal Medicine, Medical University of Silesia in Katowice.Material/Methods: Thirty-seven patients with Clostridium difficile infection diagnosed between October 2011 and November 2013 (26 months, identified among a total of 3728 patients hospitalized in this department during this period, were included in this retrospective, single-center study. The CDI definition was based on the current recommendations of the European Society of Clinical Microbiology and Infectious Diseases.Results: The observation period was divided into two 13-month intervals. Increased incidence (of borderline significance of CDI in the second period compared to the first period was observed (1.33% vs 0.65% respectively; p=0.057. Patients after kidney (n=11, kidney and pancreas (n=2 and liver (n=5 transplantation represented 48% of the analyzed CDI patients, and in half of these patients (50% CDI symptoms occurred within the first 3 months after transplantation. Clostridium difficile infection leads to irreversible deterioration of graft function in 38% of kidney recipients. Most incidents of CDI (70% were identified as nosocomial infection.Conclusions: 1. Clostridium difficile infection is particularly common among patients in the early period after solid organ transplantation. 2. Clostridium difficile infection may lead to irreversible deterioration of transplanted kidney function.

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

    Science.gov (United States)

    Kačániová, Miroslava; Vukovič, Nenad; Horská, Elena; Salamon, Ivan; Bobková, Alica; Hleba, Lukáš; Fiskelová, Martina; Vatľák, 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 μL.mL(-1)-0.39 μL.mL(-1)) and the best scavenging effect for the highest concentration (50 μL.mL(-1)) was observed. The antioxidant properties

  14. Microbial Enhanced Oil Recovery-Laboratory Experiments with a Strain of Clostridium tyrobutyricum

    DEFF Research Database (Denmark)

    Jimoh, Ismaila Adetunji

    the desired metabolic products needed for enhanced oil recovery. In this study, experiments have been performed with a strain of Clostridium tyrobutyricum. The experiments focused on salinity adaptation, gas production and the ability of microbes to modify rock properties. The result of the experiments showed...... that the strain of Clostridium tyrobutyricum adapted to 10, 30, 50, and 90 g/l before the start of the experiments produce more gas with an increase factor of between 0.39-6.9 for the same salinity condition than the pure culture. The adaptation process also led to the production of a strain 90F which can grow...

  15. Clostridium perfringens bacteremia caused by choledocholithiasis in the absence of gallbladder stones.

    Science.gov (United States)

    Atia, Antwan; Raiyani, Tejas; Patel, Pranav; Patton, Robert; Young, Mark

    2012-10-21

    A 67-years-old male presented with periumbilical abdominal pain, fever and jaundice. His anaerobic blood culture was positive for clostridium perfringens. Computed tomogram scan of the abdomen and abdominal ultrasound showed normal gallbladder and common bile duct (CBD). Subsequently magnetic resonance cholangiopancreaticogram showed choledocholithiasis. Endoscopic retrograde cholangiopancreaticogramwith sphincterotomy and CBD stone extraction was performed. The patient progressively improved with antibiotic therapy Choledocholithiasis should be considered as a source of clostridium perfringens bacteremia especially in the setting of elevated liver enzymes with cholestatic pattern.

  16. Antibiotic treatment for Clostridium difficile-associated diarrhea in adults.

    Science.gov (United States)

    Nelson, R

    2007-07-18

    Clostridium difficile (C. difficile) is recognized as a frequent cause of antibiotic-associated diarrhea and colitis. The aim of this review is to establish the efficacy of antibiotic therapy for C. difficile-associated diarrhea (CDAD), to identify the most effective antibiotic treatment for CDAD in adults and to determine the need for stopping the causative antibiotic during therapy. MEDLINE (1966 to 2006), EMBASE (1980 to 2006), Cochrane Central Database of Controlled Trials and the Cochrane IBD Review Group Specialized Trials Register were searched using the following search terms: "pseudomembranous colitis and randomized trial"; "Clostridium difficile and randomized trial"; "antibiotic associated diarrhea and randomized trial". Only randomized, controlled trials assessing antibiotic treatment for CDAD were included in the review. Probiotic trials are excluded. The following outcomes were sought: initial resolution of diarrhea; initial conversion of stool to C. difficile cytotoxin and/or stool culture negative; recurrence of diarrhea; recurrence of fecal C. difficile cytotoxin and/or positive stool culture; patient response to cessation of prior antibiotic therapy; sepsis; emergent surgery: fecal diversion or colectomy; and death. Data were analyzed using the MetaView statistical package in Review Manager. For dichotomous outcomes, relative risks (RR) and 95% confidence intervals (CI) were derived from each study. When appropriate, the results of included studies were combined for each outcome. For dichotomous outcomes, pooled RR and 95% CI were calculated using a fixed effect model, except where significant heterogeneity was detected, at which time the random effects model was used. Data heterogeneity was calculated using MetaView. Twelve studies (total of 1157 participants) involving patients with diarrhea who recently received antibiotics for an infection other than C. difficile were included. The definition of diarrhea ranged from at least two loose stools

  17. Comparison of five assays for detection of Clostridium difficile toxin.

    Science.gov (United States)

    Chapin, Kimberle C; Dickenson, Roberta A; Wu, Fongman; Andrea, Sarah B

    2011-07-01

    Performance characteristics of five assays for detection of Clostridium difficile toxin were compared using fresh stool samples from patients with C. difficile infection (CDI). Assays were performed simultaneously and according to the manufacturers' instructions. Patients were included in the study if they exhibited clinical symptoms consistent with CDI. Nonmolecular assays included glutamate dehydrogenase antigen tests, with positive findings followed by the Premier Toxin A and B Enzyme Immunoassay (GDH/EIA), and the C. Diff Quik Chek Complete test. Molecular assays (PCR) included the BD GeneOhm Cdiff Assay, the Xpert C. difficile test, and the ProGastro Cd assay. Specimens were considered true positive if results were positive in two or more assays. For each method, the Youden index was calculated and cost-effectiveness was analyzed. Of 81 patients evaluated, 26 (32.1%) were positive for CDI. Sensitivity of the BD GeneOhm Cdiff assay, the Xpert C. difficile test, the ProGastro Cd assay, C. Diff Quik Chek Complete test, and two-step GDH/EIA was 96.2%, 96.2%, 88.5%, 61.5%, and 42.3%, respectively. Specificity of the Xpert C. difficile test was 96.4%, and for the other four assays was 100%. Compared with nonmolecular methods, molecular methods detected 34.7% more positive specimens. Assessment of performance characteristics and cost-effectiveness demonstrated that the BD GeneOhm Cdiff assay yielded the best results. While costly, the Xpert C. difficile test required limited processing and yielded rapid results. Because of discordant results, specimen processing, and extraction equipment requirements, the ProGastro Cd assay was the least favored molecular assay. The GDH/EIA method lacked sufficient sensitivity to be recommended. Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  18. Toxinotyping of Clostridium perfringens strains isolated from packed chicken portions

    Directory of Open Access Journals (Sweden)

    Maryam Poursoltani

    2014-06-01

    Full Text Available Background and Aim: Clostridium perfringens are classified into five toxin types A to E, on the basis of production of Alpha, Beta, Epsilon and Iota toxins. Some strains are able to produce enterotoxin, can cause food poisoning in human. The bacteria are able to produce NetB and TpeL toxins which are virulence factors in necrotic enteritis in poultry. The aim of this study was to determine the toxin profile of C. perfringens strains isolated from packed chicken portions using Single and Multiplex PCR assays. Materials and Methods: In a crossectional study, 180 sample of chicken portions including wing (n=50, liver (n=50, neck (n=50 and gizzard (n=30 were collected randomly and examined for C. perfringens contamination. For this purpose all of samples were cultured on the 7% sheep defibrinated blood agar, TSN and TSC culture media. All of the isolates were investigated for the presence of alpha, beta, epsilon, iota toxin and virulence (tpeL and netB genes. Results: In the present study, 6 isolates out of 180 samples, were confirmed as C. perfringens by culture and molecular methods. All of the isolates (100% were confirmed as cpa and cpb positive strains and belong to type C of C. perfringens. The netB gene was detected in 5 isolates (83.33% and tpeL gene in three isolates (50%. Conclusions: Our findings show the majority of C. perfringens in broilers are belong to type C which produce necrotic enteritis in poultry and may be transmitted to human through poultry products.

  19. Spore formation and toxin production in Clostridium difficile biofilms.

    Directory of Open Access Journals (Sweden)

    Ekaterina G Semenyuk

    Full Text Available The ability to grow as a biofilm can facilitate survival of bacteria in the environment and promote infection. To better characterize biofilm formation in the pathogen Clostridium difficile, we established a colony biofilm culture method for this organism on a polycarbonate filter, and analyzed the matrix and the cells in biofilms from a variety of clinical isolates over several days of biofilm culture. We found that biofilms readily formed in all strains analyzed, and that spores were abundant within about 6 days. We also found that extracellular DNA (eDNA, polysaccharide and protein was readily detected in the matrix of all strains, including the major toxins A and/or B, in toxigenic strains. All the strains we analyzed formed spores. Apart from strains 630 and VPI10463, which sporulated in the biofilm at relatively low frequencies, the frequencies of biofilm sporulation varied between 46 and 65%, suggesting that variations in sporulation levels among strains is unlikely to be a major factor in variation in the severity of disease. Spores in biofilms also had reduced germination efficiency compared to spores obtained by a conventional sporulation protocol. Transmission electron microscopy revealed that in 3 day-old biofilms, the outermost structure of the spore is a lightly staining coat. However, after 6 days, material that resembles cell debris in the matrix surrounds the spore, and darkly staining granules are closely associated with the spores surface. In 14 day-old biofilms, relatively few spores are surrounded by the apparent cell debris, and the surface-associated granules are present at higher density at the coat surface. Finally, we showed that biofilm cells possess 100-fold greater resistance to the antibiotic metronidazole then do cells cultured in liquid media. Taken together, our data suggest that C. difficile cells and spores in biofilms have specialized properties that may facilitate infection.

  20. Bystander Host Cell Killing Effects of Clostridium perfringens Enterotoxin

    Directory of Open Access Journals (Sweden)

    Archana Shrestha

    2016-12-01

    Full Text Available Clostridium perfringens enterotoxin (CPE binds to claudin receptors, e.g., claudin-4, and then forms a pore that triggers cell death. Pure cultures of host cells that do not express claudin receptors, e.g., fibroblasts, are unaffected by pathophysiologically relevant CPE concentrations in vitro. However, both CPE-insensitive and CPE-sensitive host cells are present in vivo. Therefore, this study tested whether CPE treatment might affect fibroblasts when cocultured with CPE-sensitive claudin-4 fibroblast transfectants or Caco-2 cells. Under these conditions, immunofluorescence microscopy detected increased death of fibroblasts. This cytotoxic effect involved release of a toxic factor from the dying CPE-sensitive cells, since it could be reproduced using culture supernatants from CPE-treated sensitive cells. Supernatants from CPE-treated sensitive cells, particularly Caco-2 cells, were found to contain high levels of membrane vesicles, often containing a CPE species. However, most cytotoxic activity remained in those supernatants even after membrane vesicle depletion, and CPE was not detected in fibroblasts treated with supernatants from CPE-treated sensitive cells. Instead, characterization studies suggest that a major cytotoxic factor present in supernatants from CPE-treated sensitive cells may be a 10- to 30-kDa host serine protease or require the action of that host serine protease. Induction of caspase-3-mediated apoptosis was found to be important for triggering release of the cytotoxic factor(s from CPE-treated sensitive host cells. Furthermore, the cytotoxic factor(s in these supernatants was shown to induce a caspase-3-mediated killing of fibroblasts. This bystander killing effect due to release of cytotoxic factors from CPE-treated sensitive cells could contribute to CPE-mediated disease.

  1. Oscillating behavior of Clostridium difficile Min proteins in Bacillus subtilis.

    Science.gov (United States)

    Makroczyová, Jana; Jamroškovič, Ján; Krascsenitsová, Eva; Labajová, Nad'a; Barák, Imrich

    2016-06-01

    In rod-shaped bacteria, the proper placement of the division septum at the midcell relies, at least partially, on the proteins of the Min system as an inhibitor of cell division. The main principle of Min system function involves the formation of an inhibitor gradient along the cell axis; however, the establishment of this gradient differs between two well-studied gram-negative and gram-positive bacteria. While in gram-negative Escherichia coli, the Min system undergoes pole-to-pole oscillation, in gram-positive Bacillus subtilis, proper spatial inhibition is achieved by the preferential attraction of the Min proteins to the cell poles. Nevertheless, when E.coli Min proteins are inserted into B.subtilis cells, they still oscillate, which negatively affects asymmetric septation during sporulation in this organism. Interestingly, homologs of both Min systems were found to be present in various combinations in the genomes of anaerobic and endospore-forming Clostridia, including the pathogenic Clostridium difficile. Here, we have investigated the localization and behavior of C.difficile Min protein homologs and showed that MinDE proteins of C.difficile can oscillate when expressed together in B.subtilis cells. We have also investigated the effects of this oscillation on B.subtilis sporulation, and observed decreased sporulation efficiency in strains harboring the MinDE genes. Additionally, we have evaluated the effects of C.difficile Min protein expression on vegetative division in this heterologous host. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  2. Diagnostic trends in Clostridium difficile detection in Finnish microbiology laboratories.

    Science.gov (United States)

    Könönen, Eija; Rasinperä, Marja; Virolainen, Anni; Mentula, Silja; Lyytikäinen, Outi

    2009-12-01

    Due to increased interest directed to Clostridium difficile-associated infections, a questionnaire survey of laboratory diagnostics of toxin-producing C. difficile was conducted in Finland in June 2006. Different aspects pertaining to C. difficile diagnosis, such as requests and criteria used for testing, methods used for its detection, yearly changes in diagnostics since 1996, and the total number of investigations positive for C. difficile in 2005, were asked in the questionnaire, which was sent to 32 clinical microbiology laboratories, including all hospital-affiliated and the relevant private clinical microbiology laboratories in Finland. The situation was updated by phone and email correspondence in September 2008. In June 2006, 28 (88%) laboratories responded to the questionnaire survey; 24 of them reported routinely testing requested stool specimens for C. difficile. Main laboratory methods included toxin detection (21/24; 88%) and/or anaerobic culture (19/24; 79%). In June 2006, 18 (86%) of the 21 laboratories detecting toxins directly from feces, from the isolate, or both used methods for both toxin A (TcdA) and B (TcdB), whereas only one laboratory did so in 1996. By September 2008, all of the 23 laboratories performing diagnostics for C. difficile used methods for both TcdA and TcdB. In 2006, the number of specimens processed per 100,000 population varied remarkably between different hospital districts. In conclusion, culturing C. difficile is common and there has been a favorable shift in toxin detection practice in Finnish clinical microbiology laboratories. However, the variability in diagnostic activity reported in 2006 creates a challenge for national monitoring of the epidemiology of C. difficile and related diseases.

  3. Enteric Diseases of Poultry with Special Attention to Clostridium perfringens

    Directory of Open Access Journals (Sweden)

    Hafez Mohamed Hafez

    2011-06-01

    Full Text Available The enteric heath of growing poultry is imperative to success of the production. The basic role of poultry production is turning feed stuffs into meat. Any changes in this turning process, due to mechanical, chemical or biological disturbance of digestive system (enteric disorders is mostly accompanied with high economic losses due to poor performance, increased mortality rates and increased medication costs. The severity of clinical signs and course of the disorders are influenced several factors such as management, nutrition and the involved agent(s. Several pathogens (viruses, bacteria and parasites are incriminated as possible cause of enteric disorders either alone (mono-causal, in synergy with other micro-organisms (multi-causal, or with non-infectious causes such as feed and /or management related factors. In addition, excessive levels of mycotoxins and biogenic amines in feed lead to enteric disorders. Also factors such as high stocking density, poor litter conditions, poor hygiene and high ammonia level and other stressful situation may reduce the resistance of the birds and increases their susceptibility to infections. Under field conditions, however, under filed conditions it is difficult to determine whether the true cause of enteric disorders, is of infectious or non-infectious origin. In recent years and since the ban of use of antimicrobial growth promoters in several countries the incidence of intestinal disorders especially those caused by clostridial infection was drastically increased. The present review described in general the several factors involved in enteric disorders and summarized the available literatures about Clostridium perfringens infection in poultry.

  4. Antimicrobial susceptibility of Clostridium difficile isolated in Thailand

    Directory of Open Access Journals (Sweden)

    Papanin Putsathit

    2017-06-01

    Full Text Available Abstract Background Exposure to antimicrobials is the major risk factor associated with Clostridium difficile infection (CDI. Paradoxically, treatment of CDI with antimicrobials remains the preferred option. To date, only three studies have investigated the antimicrobial susceptibility of C. difficile from Thailand, two of which were published in the 1990s. This study aimed to investigate the contemporary antibiotic susceptibility of C. difficile isolated from patients in Thailand. Methods A collection of 105 C. difficile isolated from inpatients admitted at Siriraj Hospital in Bangkok in 2015 was tested for their susceptibility to nine antimicrobials via an agar incorporation method. Results All isolates were susceptible to vancomycin, metronidazole, amoxicillin/clavulanate and meropenem. Resistance to clindamycin, erythromycin and moxifloxacin was observed in 73.3%, 35.2% and 21.0% of the isolates, respectively. The in vitro activity of fidaxomicin (MIC50/MIC90 0.06/0.25 mg/L was superior to first-line therapies vancomycin (MIC50/MIC90 1/2 mg/L and metronidazole (MIC50/MIC90 0.25/0.25 mg/L. Rifaximin exhibited potent activity against 85.7% of the isolates (MIC ≤0.03 mg/L, and its MIC50 (0.015 mg/L was the lowest among all antimicrobials tested. The prevalence of multi-drug resistant C. difficile, defined by resistance to ≥3 antimicrobials, was 21.9% (23/105. Conclusions A high level of resistance against multiple classes of antimicrobial was observed, emphasising the need for enhanced antimicrobial stewardship and educational programmes to effectively disseminate information regarding C. difficile awareness and appropriate use of antimicrobials to healthcare workers and the general public.

  5. Comparison of Five Assays for Detection of Clostridium difficile Toxin

    Science.gov (United States)

    Chapin, Kimberle C.; Dickenson, Roberta A.; Wu, Fongman; Andrea, Sarah B.

    2011-01-01

    Performance characteristics of five assays for detection of Clostridium difficile toxin were compared using fresh stool samples from patients with C. difficile infection (CDI). Assays were performed simultaneously and according to the manufacturers' instructions. Patients were included in the study if they exhibited clinical symptoms consistent with CDI. Nonmolecular assays included glutamate dehydrogenase antigen tests, with positive findings followed by the Premier Toxin A and B Enzyme Immunoassay (GDH/EIA), and the C. Diff Quik Chek Complete test. Molecular assays (PCR) included the BD GeneOhm Cdiff Assay, the Xpert C. difficile test, and the ProGastro Cd assay. Specimens were considered true positive if results were positive in two or more assays. For each method, the Youden index was calculated and cost-effectiveness was analyzed. Of 81 patients evaluated, 26 (32.1%) were positive for CDI. Sensitivity of the BD GeneOhm Cdiff assay, the Xpert C. difficile test, the ProGastro Cd assay, C. Diff Quik Chek Complete test, and two-step GDH/EIA was 96.2%, 96.2%, 88.5%, 61.5%, and 42.3%, respectively. Specificity of the Xpert C. difficile test was 96.4%, and for the other four assays was 100%. Compared with nonmolecular methods, molecular methods detected 34.7% more positive specimens. Assessment of performance characteristics and cost-effectiveness demonstrated that the BD GeneOhm Cdiff assay yielded the best results. While costly, the Xpert C. difficile test required limited processing and yielded rapid results. Because of discordant results, specimen processing, and extraction equipment requirements, the ProGastro Cd assay was the least favored molecular assay. The GDH/EIA method lacked sufficient sensitivity to be recommended. PMID:21704273

  6. DNA microarray-based PCR ribotyping of Clostridium difficile.

    Science.gov (United States)

    Schneeberg, Alexander; Ehricht, Ralf; Slickers, Peter; Baier, Vico; Neubauer, Heinrich; Zimmermann, Stefan; Rabold, Denise; Lübke-Becker, Antina; Seyboldt, Christian

    2015-02-01

    This study presents a DNA microarray-based assay for fast and simple PCR ribotyping of Clostridium difficile strains. Hybridization probes were designed to query the modularly structured intergenic spacer region (ISR), which is also the template for conventional and PCR ribotyping with subsequent capillary gel electrophoresis (seq-PCR) ribotyping. The probes were derived from sequences available in GenBank as well as from theoretical ISR module combinations. A database of reference hybridization patterns was set up from a collection of 142 well-characterized C. difficile isolates representing 48 seq-PCR ribotypes. The reference hybridization patterns calculated by the arithmetic mean were compared using a similarity matrix analysis. The 48 investigated seq-PCR ribotypes revealed 27 array profiles that were clearly distinguishable. The most frequent human-pathogenic ribotypes 001, 014/020, 027, and 078/126 were discriminated by the microarray. C. difficile strains related to 078/126 (033, 045/FLI01, 078, 126, 126/FLI01, 413, 413/FLI01, 598, 620, 652, and 660) and 014/020 (014, 020, and 449) showed similar hybridization patterns, confirming their genetic relatedness, which was previously reported. A panel of 50 C. difficile field isolates was tested by seq-PCR ribotyping and the DNA microarray-based assay in parallel. Taking into account that the current version of the microarray does not discriminate some closely related seq-PCR ribotypes, all isolates were typed correctly. Moreover, seq-PCR ribotypes without reference profiles available in the database (ribotype 009 and 5 new types) were correctly recognized as new ribotypes, confirming the performance and expansion potential of the microarray. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Spore formation and toxin production in Clostridium difficile biofilms.

    Science.gov (United States)

    Semenyuk, Ekaterina G; Laning, Michelle L; Foley, Jennifer; Johnston, Pehga F; Knight, Katherine L; Gerding, Dale N; Driks, Adam

    2014-01-01

    The ability to grow as a biofilm can facilitate survival of bacteria in the environment and promote infection. To better characterize biofilm formation in the pathogen Clostridium difficile, we established a colony biofilm culture method for this organism on a polycarbonate filter, and analyzed the matrix and the cells in biofilms from a variety of clinical isolates over several days of biofilm culture. We found that biofilms readily formed in all strains analyzed, and that spores were abundant within about 6 days. We also found that extracellular DNA (eDNA), polysaccharide and protein was readily detected in the matrix of all strains, including the major toxins A and/or B, in toxigenic strains. All the strains we analyzed formed spores. Apart from strains 630 and VPI10463, which sporulated in the biofilm at relatively low frequencies, the frequencies of biofilm sporulation varied between 46 and 65%, suggesting that variations in sporulation levels among strains is unlikely to be a major factor in variation in the severity of disease. Spores in biofilms also had reduced germination efficiency compared to spores obtained by a conventional sporulation protocol. Transmission electron microscopy revealed that in 3 day-old biofilms, the outermost structure of the spore is a lightly staining coat. However, after 6 days, material that resembles cell debris in the matrix surrounds the spore, and darkly staining granules are closely associated with the spores surface. In 14 day-old biofilms, relatively few spores are surrounded by the apparent cell debris, and the surface-associated granules are present at higher density at the coat surface. Finally, we showed that biofilm cells possess 100-fold greater resistance to the antibiotic metronidazole then do cells cultured in liquid media. Taken together, our data suggest that C. difficile cells and spores in biofilms have specialized properties that may facilitate infection.

  8. Clostridium difficile colonization and infection in patients with hepatic cirrhosis.

    Science.gov (United States)

    Yan, Dong; Chen, Yunbo; Lv, Tao; Huang, Yandi; Yang, Jiezuan; Li, Yongtao; Huang, Jianrong; Li, Lanjuan

    2017-10-01

    The aim of this study was to investigate the toxigenic Clostridium difficile colonization (CDC, colonization with toxigenic C. difficile but without symptoms) and C. difficile infection (CDI, active C. difficile infection resulting in disease symptoms) in hepatic cirrhosis patients, identify the risk factors of CDC, and determine the correlation between CDC and CDI. The strains of toxigenic C. difficile were isolated from patients with hepatic cirrhosis within 48 h after admission, followed by multilocus sequence typing (MLST). Patients were divided into toxigenic CDC group and noncolonized (NC) group according to the colonization. Logistic regression analysis was performed to analyse the risk factors for the CDC. Besides, the CDI incidence was compared between the two groups. Colonization of toxigenic C. difficile was identified in 104 cases (19.8 %). Eighteen sequence types (STs) were identified, among which ST-3, ST-54, ST-35 and ST-37 were the predominant types. Child-Pugh class C(relative risk, RR, 3.025; 95 % CI: 1.410-6.488), decrease of prothrombin time activity (PTA) (RR 2.180; 95 % CI: 1.368-3.476), decrease of platelet (RR 2.746; 95 % CI: 0.931-8.103) and concurrent hepatic encephalopathy (RR 1.740; 95 % CI: 1.012-2.990) were identified as the risk factors for the hepatic cirrhosis patients with CDC. The CDI incidence in the CDC group was also significantly higher than that of the NC group (26.0 % vs 1.7 %, Pdifficile colonization. Child's class C, decrease of PTA and platelet, and concurrent hepatic encephalopathy were the risk factors for the hepatic cirrhosis patients with C. difficile colonization. Hepatic cirrhosis patients with C. difficile colonization were more susceptible to CDI.

  9. Overdiagnosis of Clostridium difficile Infection in the Molecular Test Era.

    Science.gov (United States)

    Polage, Christopher R; Gyorke, Clare E; Kennedy, Michael A; Leslie, Jhansi L; Chin, David L; Wang, Susan; Nguyen, Hien H; Huang, Bin; Tang, Yi-Wei; Lee, Lenora W; Kim, Kyoungmi; Taylor, Sandra; Romano, Patrick S; Panacek, Edward A; Goodell, Parker B; Solnick, Jay V; Cohen, Stuart H

    2015-11-01

    Clostridium difficile is a major cause of health care-associated infection, but disagreement between diagnostic tests is an ongoing barrier to clinical decision making and public health reporting. Molecular tests are increasingly used to diagnose C difficile infection (CDI), but many molecular test-positive patients lack toxins that historically defined disease, making it unclear if they need treatment. To determine the natural history and need for treatment of patients who are toxin immunoassay negative and polymerase chain reaction (PCR) positive (Tox-/PCR+) for CDI. Prospective observational cohort study at a single academic medical center among 1416 hospitalized adults tested for C difficile toxins 72 hours or longer after admission between December 1, 2010, and October 20, 2012. The analysis was conducted in stages with revisions from April 27, 2013, to January 13, 2015. Patients undergoing C difficile testing were grouped by US Food and Drug Administration-approved toxin and PCR tests as Tox+/PCR+, Tox-/PCR+, or Tox-/PCR-. Toxin results were reported clinically. Polymerase chain reaction results were not reported. The main study outcomes were duration of diarrhea during up to 14 days of treatment, rate of CDI-related complications (ie, colectomy, megacolon, or intensive care unit care) and CDI-related death within 30 days. Twenty-one percent (293 of 1416) of hospitalized adults tested for C difficile were positive by PCR, but 44.7% (131 of 293) had toxins detected by the clinical toxin test. At baseline, Tox-/PCR+ patients had lower C difficile bacterial load and less antibiotic exposure, fecal inflammation, and diarrhea than Tox+/PCR+ patients (P difficile by either method. Exclusive reliance on molecular tests for CDI diagnosis without tests for toxins or host response is likely to result in overdiagnosis, overtreatment, and increased health care costs.

  10. Fate of ingested Clostridium difficile spores in mice.

    Directory of Open Access Journals (Sweden)

    Amber Howerton

    Full Text Available Clostridium difficile infection (CDI is a leading cause of antibiotic-associated diarrhea, a major nosocomial complication. The infective form of C. difficile is the spore, a dormant and resistant structure that forms under stress. Although spore germination is the first committed step in CDI onset, the temporal and spatial distribution of ingested C. difficile spores is not clearly understood. We recently reported that CamSA, a synthetic bile salt analog, inhibits C. difficile spore germination in vitro and in vivo. In this study, we took advantage of the anti-germination activity of bile salts to determine the fate of ingested C. difficile spores. We tested four different bile salts for efficacy in preventing CDI. Since CamSA was the only anti-germinant tested able to prevent signs of CDI, we characterized CamSa's in vitro stability, distribution, and cytotoxicity. We report that CamSA is stable to simulated gastrointestinal (GI environments, but will be degraded by members of the natural microbiota found in a healthy gut. Our data suggest that CamSA will not be systemically available, but instead will be localized to the GI tract. Since in vitro pharmacological parameters were acceptable, CamSA was used to probe the mouse model of CDI. By varying the timing of CamSA dosage, we estimated that C. difficile spores germinated and established infection less than 10 hours after ingestion. We also showed that ingested C. difficile spores rapidly transited through the GI tract and accumulated in the colon and cecum of CamSA-treated mice. From there, C. difficile spores were slowly shed over a 96-hour period. To our knowledge, this is the first report of using molecular probes to obtain disease progression information for C. difficile infection.

  11. The Burden of Clostridium difficile after Cervical Spine Surgery.

    Science.gov (United States)

    Guzman, Javier Z; Skovrlj, Branko; Rothenberg, Edward S; Lu, Young; McAnany, Steven; Cho, Samuel K; Hecht, Andrew C; Qureshi, Sheeraz A

    2016-06-01

    Study Design Retrospective database analysis. Objective The purpose of this study is to investigate incidence, comorbidities, and impact on health care resources of Clostridium difficile infection after cervical spine surgery. Methods A total of 1,602,130 cervical spine surgeries from the Nationwide Inpatient Sample database from 2002 to 2011 were included. Patients were included for study based on International Classification of Diseases Ninth Revision, Clinical Modification procedural codes for cervical spine surgery for degenerative spine diagnoses. Baseline patient characteristics were determined. Multivariable analyses assessed factors associated with increased incidence of C. difficile and risk of mortality. Results Incidence of C. difficile infection in postoperative cervical spine surgery hospitalizations is 0.08%, significantly increased since 2002 (p difficile infection were significantly increased in patients with comorbidities such as congestive heart failure, renal failure, and perivascular disease. Circumferential cervical fusion (odds ratio [OR] = 2.93, p difficile infection after degenerative cervical spine surgery. C. difficile infection after cervical spine surgery results in extended length of stay (p costs (p difficile after cervical spine surgery is nearly 8% versus 0.19% otherwise (p difficile to be a significant predictor of inpatient mortality (OR = 3.99, p difficile increases the risk of in-hospital mortality and costs approximately $6,830,695 per year to manage in patients undergoing elective cervical spine surgery. Patients with comorbidities such as renal failure or congestive heart failure have increased probability of developing infection after surgery. Accepted antibiotic guidelines in this population must be followed to decrease the risk of developing postoperative C. difficile colitis.

  12. Structural insight into the Clostridium difficile ethanolamine utilisation microcompartment.

    Directory of Open Access Journals (Sweden)

    Alison C Pitts

    Full Text Available Bacterial microcompartments form a protective proteinaceous barrier around metabolic enzymes that process unstable or toxic chemical intermediates. The genome of the virulent, multidrug-resistant Clostridium difficile 630 strain contains an operon, eut, encoding a bacterial microcompartment with genes for the breakdown of ethanolamine and its utilisation as a source of reduced nitrogen and carbon. The C. difficile eut operon displays regulatory genetic elements and protein encoding regions in common with homologous loci found in the genomes of other bacteria, including the enteric pathogens Salmonella enterica and Enterococcus faecalis. The crystal structures of two microcompartment shell proteins, CD1908 and CD1918, and an uncharacterised protein with potential enzymatic activity, CD1925, were determined by X-ray crystallography. CD1908 and CD1918 display the same protein fold, though the order of secondary structure elements is permuted in CD1908 and this protein displays an N-terminal β-strand extension. These proteins form hexamers with molecules related by crystallographic and non-crystallographic symmetry. The structure of CD1925 has a cupin β-barrel fold and a putative active site that is distinct from the metal-ion dependent catalytic cupins. Thin-section transmission electron microscopy of Escherichia coli over-expressing eut proteins indicates that CD1918 is capable of self-association into arrays, suggesting an organisational role for CD1918 in the formation of this microcompartment. The work presented provides the basis for further study of the architecture and function of the C. difficile eut microcompartment, its role in metabolism and the wider consequences of intestinal colonisation and virulence in this pathogen.

  13. Genetic characterization of type A enterotoxigenic Clostridium perfringens strains.

    Directory of Open Access Journals (Sweden)

    Agi Deguchi

    2009-05-01

    Full Text Available Clostridium perfringens type A, is both a ubiquitous environmental bacterium and a major cause of human gastrointestinal disease, which usually involves strains producing C. perfringens enterotoxin (CPE. The gene (cpe encoding this toxin can be carried on the chromosome or a large plasmid. Interestingly, strains carrying cpe on the chromosome and strains carrying cpe on a plasmid often exhibit different biological characteristics, such as resistance properties against heat. In this study, we investigated the genetic properties of C. perfringens by PCR-surveying 21 housekeeping genes and genes on representative plasmids and then confirmed those results by Southern blot assay (SB of five genes. Furthermore, sequencing analysis of eight housekeeping genes and multilocus sequence typing (MLST analysis were also performed. Fifty-eight C. perfringens strains were examined, including isolates from: food poisoning cases, human gastrointestinal disease cases, foods in Japan or the USA, or feces of healthy humans. In the PCR survey, eight of eleven housekeeping genes amplified positive reactions in all strains tested. However, by PCR survey and SB assay, one representative virulence gene, pfoA, was not detected in any strains carrying cpe on the chromosome. Genes involved in conjugative transfer of the cpe plasmid were also absent from almost all chromosomal cpe strains. MLST showed that, regardless of their geographic origin, date of isolation, or isolation source, chromosomal cpe isolates, i assemble into one definitive cluster ii lack pfoA and iii lack a plasmid related to the cpe plasmid. Similarly, independent of their origin, strains carrying a cpe plasmid also appear to be related, but are more variable than chromosomal cpe strains, possibly because of the instability of cpe-borne plasmid(s and/or the conjugative transfer of cpe-plasmid(s into unrelated C. perfringens strains.

  14. Sample preparation

    International Nuclear Information System (INIS)

    Anon.

    1992-01-01

    Sample preparation prior to HPLC analysis is certainly one of the most important steps to consider in trace or ultratrace analysis. For many years scientists have tried to simplify the sample preparation process. It is rarely possible to inject a neat liquid sample or a sample where preparation may not be any more complex than dissolution of the sample in a given solvent. The last process alone can remove insoluble materials, which is especially helpful with the samples in complex matrices if other interactions do not affect extraction. Here, it is very likely a large number of components will not dissolve and are, therefore, eliminated by a simple filtration process. In most cases, the process of sample preparation is not as simple as dissolution of the component interest. At times, enrichment is necessary, that is, the component of interest is present in very large volume or mass of material. It needs to be concentrated in some manner so a small volume of the concentrated or enriched sample can be injected into HPLC. 88 refs

  15. Factores de riesgo para la infección por Clostridium difficile Risk factors for Clostridium difficileinfection

    Directory of Open Access Journals (Sweden)

    María Gabriela Becerra

    2011-12-01

    Full Text Available Introducción. Clostridium difficile es un bacilo Gram positivo, anaerobio estricto y formador de esporas, capaz de persistir bajo condiciones adversas durante mucho tiempo. Es una de las causas más frecuentes de infección asociada a la atención en salud, cuyas manifestaciones clínicas van desde diarrea sin complicaciones hasta sepsis e, incluso, la muerte. El propósito de este estudio fue determinar los factores de riesgo para infección por C. difficile en un hospital universitario. Materiales y métodos. Se llevó a cabo un estudio de casos y controles de pacientes mayores de 18 años, de ambos sexos, que presentaron diarrea durante su hospitalización en el Hospital Universitario de San Vicente Fundación y a quienes se les realizó la prueba para la detección de las toxina A y B de C. difficile, entre septiembre de 2009 y diciembre de 2010. A partir de esta población se seleccionaron 22 casos y 44 controles. Resultados. Los factores de riesgo que se encontraron asociados fueron: edad mayor de 65 años (OR=3,4; IC95% 1,1-10,1; p=0,05, la estancia en unidad de cuidados intensivos (OR=4,0; IC95% 1,3-12,2; p=0,02 y el uso de inhibidores de la bomba de protones (OR=5,15; IC95% 1,6-15,9; pIntroduction: Clostridium difficile is a Gram positive strict anaerobic spore-forming bacillus, so it is able to persist under adverse conditions for long time. This microorganism is a the most common cause of health care associated infection, with clinical manifestations ranging from uncomplicated diarrhea to sepsis and even death. The purpose of this study was to determine the risk factors for C. difficile infection in a teaching hospital. Materials and methods: We conducted a case control study in patients over 18 years, of both gender, who had developed diarrhea during their hospitalization in teaching Hospital of San Vicente Fundación and who underwent the toxin test for C. difficile, between September 2009 and December 2010. From this population we

  16. Clostridium Bacteria and Autism Spectrum Conditions: A Systematic Review and Hypothetical Contribution of Environmental Glyphosate Levels

    Directory of Open Access Journals (Sweden)

    Isadora Argou-Cardozo

    2018-04-01

    Full Text Available Nowadays, there seems to be a consensus about the multifactorial nature of autism spectrum disorders (ASD. The literature provides hypotheses dealing with numerous environmental factors and genes accounting for the apparently higher prevalence of this condition. Researchers have shown evidence regarding the impact of gut bacteria on neurological outcomes, altering behavior and potentially affecting the onset and/or severity of psychiatric disorders. Pesticides and agrotoxics are also included among this long list of ASD-related environmental stressors. Of note, ingestion of glyphosate (GLY, a broad-spectrum systemic herbicide, can reduce beneficial bacteria in the gastrointestinal tract microbiota without exerting any effects on the Clostridium population, which is highly resistant to this herbicide. In the present study, (i we performed a systematic review to evaluate the relationship between Clostridium bacteria and the probability of developing and/or aggravating autism among children. For that purpose, electronic searches were performed on Medline/PubMed and Scielo databases for identification of relevant studies published in English up to December 2017. Two independent researches selected the studies and analyzed the data. The results of the present systematic review demonstrate an interrelation between Clostridium bacteria colonization of the intestinal tract and autism. Finally, (ii we also hypothesize about how environmental GLY levels may deleteriously influence the gut–brain axis by boosting the growth of Clostridium bacteria in autistic toddlers.

  17. Vermin on pig farms are vectors for Clostridium difficile PCR ribotypes 078 and 045

    NARCIS (Netherlands)

    Burt, S.A.; Siemeling, L.; Kuijper, E.J.; Lipman, L.J.A.

    2012-01-01

    Clostridium difficile is a gram positive, spore forming, toxin producing, anaerobic bacteria and an opportunistic pathogen for Man and many animal species, causing diarrhea in young piglets. Piglets probably become colonized from the environment. To investigate the possible spread and transmission

  18. The impact of hospital-onset Clostridium difficile infection on outcomes of hospitalized patients with sepsis.

    Science.gov (United States)

    Lagu, Tara; Stefan, Mihaela S; Haessler, Sarah; Higgins, Thomas L; Rothberg, Michael B; Nathanson, Brian H; Hannon, Nicholas S; Steingrub, Jay S; Lindenauer, Peter K

    2014-07-01

    To examine the impact of hospital-onset Clostridium difficile infection (HOCDI) on the outcomes of patients with sepsis. Most prior studies that have addressed this issue lacked adequate matching to controls, suffered from small sample size, or failed to consider time to infection. Retrospective cohort study. We identified adults with a principal or secondary diagnosis of sepsis who received care at 1 of the institutions that participated in a large multihospital database between July 1, 2004 and December 31, 2010. Among eligible patients with sepsis, we identified patients who developed HOCDI during their hospital stay. We used propensity matching and date of diagnosis to match cases to patients without Clostridium difficile infections and compared outcomes between the 2 groups. Of 218,915 sepsis patients, 2368 (1.08%) developed HOCDI. Unadjusted in-hospital mortality was significantly higher in HOCDI patients than controls (25% vs 10%, P Clostridium difficile infections was 5.1 days longer than controls (95% confidence interval: 4.4-5.8) and the median-adjusted cost increase was $4916 (P Clostridium difficile infection was associated with increased mortality, LOS, and cost. Our results can be used to assess the cost-effectiveness of prevention programs and suggest that efforts directed toward high-risk patient populations are needed. © 2014 Society of Hospital Medicine.

  19. Susceptibility of Clostridium difficile Toward Antimicrobial Agents Used as Feed Additives for Food Animals

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Tvede, Michael

    2011-01-01

    A total of 65 toxigenic Clostridium difficile strains isolated from patients with antibiotic-associated diarrhea were tested for susceptibility to avilamycin, flavomycin, monensin, and salinomycin. Except for flavomycin the substances showed in vitro efficacy comparable to reports of the currentl...

  20. Flooding and Health Care Visits for Clostridium Difficile Infection: A Case-Crossover Analysis

    Science.gov (United States)

    Floods can contaminate potable water and other resources, thus increasing the potential for fecal-oral transmission of pathogens. Clostridium difficile is a bacterium that can spread by water and cause acute gastrointestinal illness. It often affects older adults who are hospital...

  1. Increase in Clostridium difficile-related Mortality Rates, United States, 1999-2004

    Centers for Disease Control (CDC) Podcasts

    2008-01-08

    Deaths related to Clostridium difficile are on the rise in the United States. Matthew Redelings from the Los Angeles County Department of Health discusses the increase and what can be done to prevent this infection.  Created: 1/8/2008 by Emerging Infectious Diseases.   Date Released: 1/8/2008.

  2. FERMENTATION OF INULIN BY A NEW STRAIN OF CLOSTRIDIUM-THERMOAUTOTROPHICUM ISOLATED FROM DAHLIA TUBERS

    NARCIS (Netherlands)

    DRENT, WJ; GOTTSCHAL, JC

    1991-01-01

    A new inulin-fermenting strain of Clostridium thermoautotrophicum was isolated through enrichment on dahlia tubers, and subsequent plating on agar media with undissolved inulin. Both the cell-bound and cell-free inulinase(s) functioned optimally at 60-degrees-C and at neutral pH. This new strain I1

  3. Suppression by Saccharomyces boulardii of toxigenic Clostridium difficile overgrowth after vancomycin treatment in hamsters.

    Science.gov (United States)

    Elmer, G W; McFarland, L V

    1987-01-01

    Saccharomyces boulardii prevented the development of high counts of Clostridium difficile, high titers of toxin B, and positive latex agglutination tests after cessation of vancomycin treatment for hamsters. The protocol used was designed to stimulate relapse of human C. difficile-associated colitis. S. boulardii was protective in this model. PMID:3566236

  4. Carboxymethyl cellulase and cellobiase production by Clostridium acetobutylicum in an industrial fermentation medium.

    OpenAIRE

    Allcock, E R; Woods, D R

    1981-01-01

    The production of a carboxymethyl cellulase and a cellobiase by Clostridium acetobutylicum was demonstrated. In liquid medium the carboxymethyl cellulase was induced by molasses, and it was not repressed by glucose. Optimum carboxymethyl cellulase activity occurred at pH 4.6 and 37 degrees C.

  5. Clostridium beijerinckii cells expressing Neocallimastix patriciarum glycoside hydrolases show enhanced Lichenan utilization and solvent production

    NARCIS (Netherlands)

    Lopez-Contreras, A.; Oost, van der J.; Claassen, P.; Mooibroek, H.; Vos, de W.M.

    2001-01-01

    Growth and the production of acetone, butanol, and ethanol by Clostridium beijerinckii NCIMB 8052 on several polysaccharides and sugars were analyzed. On crystalline cellulose, growth and solvent production were observed only when a mixture of fungal cellulases was added to the medium. On lichenan

  6. Trends and Seasonality in Antibiotic Resistance Among Elderly Patients with Clostridium Difficile-Associated Disease

    Science.gov (United States)

    In the US, over 300,000 cases of Clostridium dijficile-associated disease (CDAD) occur annually in hospitals or long-term care facilities and incidence has risen over the past two decades potentially due to increased antibiotic use. A primary risk factor for CDAD is previous anti...

  7. Clostridium perfringens infection complicating periprosthetic fracture fixation about the hip: successful treatment with early aggressive debridement.

    LENUS (Irish Health Repository)

    Baker, Joseph F

    2012-07-13

    Periprosthetic fracture and infection are both challenges following hip arthroplasty. We report the case of an 87 year old female who underwent open reduction and internal fixation of a periprosthetic femoral fracture. Her post-operative course was complicated by infection with Clostridium perfringens. Early aggressive antibiotic treatment and surgical debridement were successful, and allowed retention of the original components.

  8. Isolation and molecular characterization of Clostridium perfringens from healthy Merino lambs in Patagonia region, Argentina

    DEFF Research Database (Denmark)

    Mignaqui, A. C.; Marcellino, R. B.; Ronco, Troels

    2017-01-01

    The presence and molecular characterization of Clostridium perfringens in healthy Merino lambs over a six-month period was investigated in this study. Overall, a high prevalence of C. perfringens was detected, even in day-old lambs. Even though the majority of the isolates were characterized...

  9. Clostridium perfringens bacteremia caused by choledocholithiasis in the absence of gallbladder stones

    OpenAIRE

    Atia, Antwan; Raiyani, Tejas; Patel, Pranav; Patton, Robert; Young, Mark

    2012-01-01

    A 67-years-old male presented with periumbilical abdominal pain, fever and jaundice. His anaerobic blood culture was positive for clostridium perfringens. Computed tomogram scan of the abdomen and abdominal ultrasound showed normal gallbladder and common bile duct (CBD). Subsequently magnetic resonance cholangiopancreaticogram showed choledocholithiasis. Endoscopic retrograde cholangiopancreaticogramwith sphincterotomy and CBD stone extraction was performed. The patient progressively improved...

  10. Substrate-induced production and secretion of cellulases by Clostridium acetobutylicum

    NARCIS (Netherlands)

    Lopez Contreras, A.M.; Gabor, K.; Martens, A.A.; Renckens, B.A.M.; Claassen, P.A.M.; Oost, van der J.; Vos, de W.M.

    2004-01-01

    Clostridium acetobutylicum ATCC 824 is a solventogenic bacterium that grows heterotrophically on a variety of carbohydrates, including glucose, cellobiose, xylose, and lichenan, a linear polymer of beta-1,3- and beta-1,4-linked beta-D-glucose units. C. acetobutylicum does not degrade cellulose,

  11. Continuous xylose fermentation by Clostridium acetobutylicum – Kinetics and energetics issues under acidogenesis conditions

    NARCIS (Netherlands)

    Procentese, A.; Raganati, F.; Olivieri, G.; Russo, M.E.; Salatino, P.; Marzocchella, A.

    2014-01-01

    The paper reports the assessment of the growth kinetics of Clostridium acetobutylicum DSM 792 adopting xylose as carbon source. Xylose is the fundamental component of hemicellulose hydrolysis, a relevant fraction of lignocellulosic feedstocks for biofuel production. Tests were carried out in a CSTR

  12. Neurotoxin gene profiling of Clostridium botulinum types C and D gathered from different countries within Europe

    NARCIS (Netherlands)

    Woudstra, C.; Skarin, A.; Anniballi, F.; Fenicia, F.; Bano, L.; Drigo, I.; Koene, M.G.J.; Bäyon-Auboyer, M.H.; Buffereau, J.P.; Medici, D.; Fach, P.

    2012-01-01

    Clostridium botulinum types C and D, as well as their mosaic variants C-D and D-C, are associated with avian and mammalian botulism. This study reports on the development of low-density macroarrays based on the GeneDisc cycler platform (Pall-GeneDisc Technologies) applied to the simultaneous

  13. Clostridium sordellii as a Cause of Fatal Septic Shock in a Child with Hemolytic Uremic Syndrome

    Directory of Open Access Journals (Sweden)

    Rebekah Beyers

    2014-01-01

    Full Text Available Clostridium sordellii is a toxin producing ubiquitous gram-positive anaerobe, mainly associated with trauma, soft tissue skin infections, and gynecologic infection. We report a unique case of a new strain of Clostridium sordellii (not present in the Center for Disease Control (CDC database infection induced toxic shock syndrome in a previously healthy two-year-old male with colitis-related hemolytic uremic syndrome (HUS. The patient presented with dehydration, vomiting, and bloody diarrhea. He was transferred to the pediatric critical care unit (PICU for initiation of peritoneal dialysis (PD. Due to increased edema and intolerance of PD, he was transitioned to hemodialysis through a femoral vascular catheter. He subsequently developed severe septic shock with persistent leukocytosis and hypotension, resulting in subsequent death. Stool culture confirmed Shiga toxin producing Escherichia coli 0157:H7. A blood culture was positively identified for Clostridium sordellii. Clostridium sordelli is rarely reported in children; to our knowledge this is the first case described in a pediatric patient with HUS.

  14. Amino Acid Transport in the Thermophilic Anaerobe Clostridium fervidus Is Driven by an Electrochemical Sodium Gradient

    NARCIS (Netherlands)

    SPEELMANS, G; POOLMAN, B; KONINGS, WN

    Amino acid transport was studied in membranes of the peptidolytic, thermophitic, anaerobic bacterium Clostridium fervidus. Uptake of the negatively charged amino acid L-glutamate, the neutral amino acid L-serine, and the positively charged amino acid L-arginine was examined in membrane vesicles

  15. Prevention of Clostridium difficile Infection with Saccharomyces boulardii: A Systematic Review

    Directory of Open Access Journals (Sweden)

    Jennifer M Tung

    2009-01-01

    Full Text Available BACKGROUND: Clostridium difficile is a major cause of antibioticassociated diarrhea within the hospital setting. The yeast Saccharomyces boulardii has been found to have some effect in reducing the risk of C difficile infection (CDI; however, its role in preventive therapy has yet to be firmly established.

  16. The HtrA-Like Protease CD3284 Modulates Virulence of Clostridium difficile

    NARCIS (Netherlands)

    Bakker, Dennis; Buckley, Anthony M.; de Jong, Anne; van Winden, Vincent J. C.; Verhoeks, Joost P. A.; Kuipers, Oscar P.; Douce, Gillian R.; Kuijper, Ed J.; Smits, Wiep Klaas; Corver, Jeroen

    2014-01-01

    In the past decade, Clostridium difficile has emerged as an important gut pathogen. Symptoms of C. difficile infection range from mild diarrhea to pseudomembranous colitis. Besides the two main virulence factors toxin A and toxin B, other virulence factors are likely to play a role in the

  17. Community-acquired Clostridium difficile infection in children: A retrospective study.

    Science.gov (United States)

    Borali, Elena; Ortisi, Giuseppe; Moretti, Chiara; Stacul, Elisabetta Francesca; Lipreri, Rita; Gesu, Giovanni Pietro; De Giacomo, Costantino

    2015-10-01

    Community acquired-Clostridium difficile infection (CDI) has increased also in children in the last years. To determine the incidence of community-acquired CDI and to understand whether Clostridium difficile could be considered a symptom-triggering pathogen in infants. A five-year retrospective analysis (January 2007-December 2011) of faecal specimens from 124 children hospitalized in the Niguarda Ca' Granda Hospital for prolonged or muco-haemorrhagic diarrhoea was carried out. Stool samples were evaluated for common infective causes of diarrhoea and for Clostridium difficile toxins. Patients with and without CDI were compared for clinical characteristics and known risk factors for infection. Twenty-two children with CDI were identified in 5 years. An increased incidence of community-acquired CDI was observed, ranging from 0.75 per 1000 hospitalizations in 2007 to 9.8 per 1000 hospitalizations in 2011. Antimicrobial treatment was successful in all 19 children in whom it was administered; 8/22 CDI-positive children were younger than 2 years. No statistically significant differences in clinical presentation were observed between patients with and without CDI, nor in patients with and without risk factors for CDI. Our study shows that Clostridium difficile infection is increasing and suggests a possible pathogenic role in the first 2 years of life. Copyright © 2015 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

  18. The potential beneficial role of faecal microbiota transplantation in diseases other than Clostridium difficile infection

    NARCIS (Netherlands)

    Singh, R.; Nieuwdorp, M.; ten Berge, I. J. M.; Bemelman, F. J.; Geerlings, S. E.

    2014-01-01

    This review gives an outline of the indications for faecal microbiota transplantation (FMT) for diseases other than Clostridium difficile (C. difficile) infection. The remarkable efficacy of FMT against C. difficile infection has already been demonstrated. The use of FMT for other diseases, such as

  19. Tryptophan catabolism restricts IFN-γ-expressing neutrophils and Clostridium difficile immunopathology

    NARCIS (Netherlands)

    El-Zaatari, Mohamad; Chang, Yu-Ming; Zhang, Min; Franz, Matthew; Shreiner, Andrew; McDermott, Andrew J.; van der Sluijs, Koenraad F.; Lutter, René; Grasberger, Helmut; Kamada, Nobuhiko; Young, Vincent B.; Huffnagle, Gary B.; Kao, John Y.

    2014-01-01

    The interplay between Clostridium difficile and the host's metabolome is believed to influence the severity of infection. However, the mechanism for this phenomenon remains unclear. In this study, we model one of these metabolic pathways by focusing on tryptophan metabolism in the host. We found

  20. Predicting a complicated course of Clostridium difficile infection at the bedside

    NARCIS (Netherlands)

    Hensgens, M. P. M.; Dekkers, O. M.; Goorhuis, A.; LeCessie, S.; Kuijper, E. J.

    2014-01-01

    Clostridium difficile infections (CDIs) are a common cause of antibiotic-associated diarrhoea and associated with CDI-related mortality in c. 10%. To date, there is no prediction model in use that guides clinicians to identify patients at high risk for complicated CDI. From 2006 to 2009, nine Dutch

  1. Outbreak of Clostridium difficile 027 in North Zealand, Denmark, 2008-2009

    DEFF Research Database (Denmark)

    Bacci, S; St-Martin, G; Olesen, B

    2009-01-01

    We report an outbreak of Clostridium difficile PCR ribotype 027 in Denmark. The outbreak includes to date 73 cases from the area north of Copenhagen, but there may be related cases elsewhere in Zealand. Most infections are healthcare-associated and in patients who previously received antibiotic...

  2. Clostridium Bacteria and Autism Spectrum Conditions: A Systematic Review and Hypothetical Contribution of Environmental Glyphosate Levels.

    Science.gov (United States)

    Argou-Cardozo, Isadora; Zeidán-Chuliá, Fares

    2018-04-04

    Nowadays, there seems to be a consensus about the multifactorial nature of autism spectrum disorders (ASD). The literature provides hypotheses dealing with numerous environmental factors and genes accounting for the apparently higher prevalence of this condition. Researchers have shown evidence regarding the impact of gut bacteria on neurological outcomes, altering behavior and potentially affecting the onset and/or severity of psychiatric disorders. Pesticides and agrotoxics are also included among this long list of ASD-related environmental stressors. Of note, ingestion of glyphosate (GLY), a broad-spectrum systemic herbicide, can reduce beneficial bacteria in the gastrointestinal tract microbiota without exerting any effects on the Clostridium population, which is highly resistant to this herbicide. In the present study, (i) we performed a systematic review to evaluate the relationship between Clostridium bacteria and the probability of developing and/or aggravating autism among children. For that purpose, electronic searches were performed on Medline/PubMed and Scielo databases for identification of relevant studies published in English up to December 2017. Two independent researches selected the studies and analyzed the data. The results of the present systematic review demonstrate an interrelation between Clostridium bacteria colonization of the intestinal tract and autism. Finally, (ii) we also hypothesize about how environmental GLY levels may deleteriously influence the gut-brain axis by boosting the growth of Clostridium bacteria in autistic toddlers.

  3. In vitro Clostridium perfringens and Escherichia coli toxin adsorption of Varium

    Science.gov (United States)

    Enteric disease agents, such as Clostridium perfringens and Escherichia coli, produce detrimental biotoxins that cause significant economic loss annually in the poultry industry. The objective of this study was to determine the in vitro biotoxin adsorption capability of Varium. An enzyme-linked im...

  4. Constructing identities in the media: newspaper coverage analysis of a major UK Clostridium difficile outbreak.

    Science.gov (United States)

    Burnett, Emma; Johnston, Bridget; Corlett, Joanne; Kearney, Nora

    2014-07-01

    To examine how a major Clostridium difficile outbreak in the UK was represented in the media. Clostridium difficile is a serious health care-associated infection with significant global prevalence. As major outbreaks have continued to occur worldwide over the last few decades, it has also resulted in increasing media coverage. Newspaper journalists are, however, frequently criticized for sensationalized and inaccurate reporting and alarming the public. Despite such criticisms, nothing is known about how the media frame Clostridium difficile related coverage. Qualitative interpretive descriptive study. An interpretive analysis of newspaper articles from the national press that reported about the outbreak from the first day of coverage over 3 weeks (12 June-3 July 2008). Twenty-eight newspaper articles were included in the study from tabloids, broadsheets, a regional and a Sunday newspaper. Monster and war metaphors were frequently adopted to portray the severity of Clostridium difficile and the impact it can have on patient safety. In addition, the positioning of the affected patients, their families, healthcare professionals and the Government produced representations of victims, villains and heroes. This subsequently evoked notions of vulnerability, blame and conflict. The media are and will remain critical convectors of public information and, as such, are hugely influential in risk perceptions and responses. Rather than simply dismissing media coverage, further understanding around how such stories in specific contexts are constructed and represented is needed so that it can help inform future communication and management strategies. © 2013 John Wiley & Sons Ltd.

  5. Spread and epidemiology of Clostridium difficile polymerase chain reaction ribotype 027/toxinotype III in The Netherlands

    NARCIS (Netherlands)

    Goorhuis, A.; van der Kooi, T.; Vaessen, N.; Dekker, F. W.; van den Berg, R.; Harmanus, C.; van den Hof, S.; Notermans, D. W.; Kuijper, E. J.

    2007-01-01

    After reports of emerging outbreaks in Canada and the United States, Clostridium difficile-associated disease (CDAD) due to polymerase chain reaction ribotype 027 was detected in 2 medium-to-large hospitals in The Netherlands in 2005. National surveillance was initiated to investigate the spread and

  6. Emergence of Clostridium difficile infection due to a new hypervirulent strain, polymerase chain reaction ribotype 078

    NARCIS (Netherlands)

    Goorhuis, Abraham; Bakker, Dennis; Corver, Jeroen; Debast, Sylvia B.; Harmanus, Celine; Notermans, Daan W.; Bergwerff, Aldert A.; Dekker, Frido W.; Kuijper, Ed J.

    2008-01-01

    Since 2005, an increase in the prevalence of Clostridium difficile infection (CDI) due to polymerase chain reaction ribotype 078 has been noticed in The Netherlands. This strain has also been identified as the predominant strain in pigs and calves. CDI caused by type 078 was studied in relation to

  7. Effect of low-dose gamma irradiation on enterotoxin-positive strains of Clostridium perfringens Type A

    International Nuclear Information System (INIS)

    Barnhart, H.M. Jr.

    1976-01-01

    The effect of low-dose gamma irradiation on selected enterotoxin producing strains of Clostridium perfringens Type A was studied. The radioresistance of three strains NCTC-8239, NCTC-10239 and NCTC-8798 in 0.1 percent peptone water, beef gravy and ground beef was determined for both vegetative cells and spores. D 10 values were approximately 30 Krad in 0.1 percent peptone water and 175 Krad in beef menstruums. D 10 values for spores were approximately 250 Krad in 0.1 percent peptone water and 335 Krad in beef. Low-level irradiation induced a 2 hr lag for cell recovery at 37 0 C following irradiation though this was strain dependent. Heat resistance of vegetative cells decreased following irradiation, although one strain was stimulated in growth response and unaltered in its heat resistance. Spore activation and germination were not affected by low-level irradiation. Spores were not significantly inactivated at this level. Irradiation had no effect on subsequent survival of vegetative cells stored at cold temperatures. Enterotoxin production by irradiated cultures was not affected by the irradiation treatment. A method for quantitating C. perfringens enterotoxin using crossed-immunoelectrophoresis was developed. It was found that this technique could detect at least .05 g of enterotoxin, could utilize crude enterotoxin preparations and was more sensitive than other methods based on biological activity

  8. Clostridium punense sp. nov., an obligate anaerobe isolated from healthy human faeces.

    Science.gov (United States)

    Lanjekar, Vikram Bholanath; Marathe, Nachiket Prakash; Shouche, Yogesh Shreepad; Ranade, Dilip Ramchandra

    2015-12-01

    An obligately anaerobic, rod-shaped (0.5-1.0 × 2.0-10.0 μm), Gram-stain-positive bacterium, occurring mainly singly or in pairs, and designated BLPYG-8T, was isolated from faeces of a healthy human volunteer aged 56 years. Cells were non-motile. Oval, terminal spores were formed that swell the cells. The strain was affiliated with the genus Clostridium sensu stricto (Clostridium rRNA cluster I) as revealed by 16S rRNA gene sequence analysis. Strain BLPYG-8T showed 97.3 to 97.4 % 16S rRNA gene sequence similarity with Clostridium sulfidigenes DSM 18982T, Clostridium subterminale DSM 6970T and Clostridium thiosulfatireducens DSM 13105T. DNA-DNA hybridization and phenotypic analysis showed that the strain was distinct from its closest relatives, C. sulfidigenes DSM 18982T, C. subterminale DSM 6970T, C. thiosulfatireducens DSM 13105T with 54.2, 53.9 and 53.3 % DNA-DNA relatedness, respectively. Strain BLPYG-8T grew in PYG broth at temperatures between 20 and 40 °C (optimum 37 °C). The strain utilized a range of amino acids as well as carbohydrates as a source of carbon and energy. Glucose fermentation resulted in the formation of volatile fatty acids mainly acetic acid, n-butyric acid and organic acids such as succinic and lactic acid. The DNA G+C content of strain BLPYG-8T was 44.1 mol%. The major fatty acids (>10 %) were C14 : 0, iso-C15 : 0, C16 : 1ω7c and C16 : 0. Phylogenetic analysis and specific phenotypic characteristics and/or DNA G+C content differentiated the strain from its closest relatives. On the basis of these data, strain BLPYG-8T represents a novel species of the genus Clostridium, for which the name Clostridium punense sp. nov. is proposed. The type strain is BLPYG-8T ( = DSM 28650T = CCUG 64195T = MCC 2737T).

  9. Determination and evaluation of the best conditions for the production of a vaccine against Clostridium chauvoei

    Directory of Open Access Journals (Sweden)

    Jorge Humberto Ossa

    2005-05-01

    Full Text Available Clostridium chauvoei is responsible for a number of diseases such as symptomatic coal (blackleg and malignant oedema in cattle, sheep and many other domestic and wild animals. Vaccines for the control of this and other livestock diseases caused by various species of clostridia are widely used. For vaccine production it is desirable that high cellular density cultures with high immunogenic power are obtained, mainly because immunity to Clostridium chauvoei is generally considered to be antibacterial rather than antitoxic. Consequently, culture conditions have to be adequately monitored and controlled. The aim of this work was to optimise the production of a vaccine against Clostridium chauvoei based on strains IRP-434, IRP 206, ATCC 10092, ATCC 11957 and LANIP, used by “Empresa Colombiana de Productos Veterinarios”, VECOL, S.A., to determine the culture medium that allows the highest cellular density. Results show that modified clostridium medium allows the highest cellular density production with average values of 31,5X108 cell/ml for the stock IRP 434 and 31,8X108 cell/ml for the stock IRP 206. Once defined the modified clostridium medium, the stock IRP 434,IRP 206 and growth time as the most suitable for optimal biomass production ( 16 – 48 hours, the lethal activity of the stock (LD50 = 106.6 , 106.8 was determined, the immunogenicity was evaluated through a potency test carried out in guinea-pigs. The presence of the flagellar protein was confirmed through the SDS-PAGE electrophoresis technique.

  10. Acetone-butanol-ethanol production from substandard and surplus dates by Egyptian native Clostridium strains.

    Science.gov (United States)

    Abd-Alla, Mohamed Hemida; Zohri, Abdel-Naser Ahmed; El-Enany, Abdel-Wahab Elsadek; Ali, Shimaa Mohamed

    2015-04-01

    One hundred and seven mesophilic isolates of Clostridium were isolated from agricultural soils cultivated with different plants in Assuit Governorate, Egypt. Eighty isolates (out of 107) showed the ability to produce ABE (Acetone, butanol and ethanol) on T6 medium ranging from 0.036 to 31.89 g/L. The highest numbers of ABE producing isolates were obtained from soil samples of potato contributing 27 isolates, followed by 18 isolates from wheat and 10 isolates from onion. On the other hand, there were three native isolates that produced ABE more than those produced by the reference isolate Clostridium acetobutylicum ATCC 824 (11.543 g/L). The three isolates were identified based on phenotypic and gene encoding 16S rRNA as Clostridium beijerinckii ASU10 (KF372577), Clostridium chauvoei ASU55 (KF372580) and Clostridium roseum ASU58 (KF372581). The highest ABE level from substandard and surplus dates was produced by C. beijerinckii ASU10 (24.07 g/L) comprising butanol 67.15% (16.16 g/L), acetone 30.73% (7.4 g/L) and ethanol 2.12% (0.51 g/L), while C. roseum ASU58 and C. chauvoei ASU55 produced ABE contributing 20.20 and 13.79 g/L, respectively. ABE production by C. acetobutylicum ATCC 824 was 15.01 g/L. This study proved that the native strains C. beijerinckii ASU10 and C. roseum ASU58 have high competitive efficacy on ABE production from economical substrate as substandard and surplus date fruits. Additionally, using this substrate without any nutritional components is considered to be a commercial substrate for desired ABE production. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. The Epidemiology and Clinical Features of Clostridium difficile Infection in Liver Transplant Recipients.

    Science.gov (United States)

    Sullivan, Timothy; Weinberg, Alan; Rana, Meenakshi; Patel, Gopi; Huprikar, Shirish

    2016-09-01

    Clostridium difficile infection (CDI) is common after liver transplantation (LT); however, few studies have examined the risk factors, clinical manifestations, and outcomes of CDI in this population. A retrospective study of adults who underwent LT between January 1, 2011, and April 4, 2013, at The Mount Sinai Hospital was conducted. Potential risk factors were evaluated via univariate and multivariable analysis to determine predictors of CDI in this population. The clinical manifestations of CDI and patient outcomes were also reviewed. Clostridium difficile infection occurred in 27 (14%) of 192 patients after LT. In multivariable analysis, CDI was associated with having a model for end-stage liver disease score of 20 or greater (hazards ratio, 2.90; 95% confidence interval, 1.29-6.52; P = 0.010), and receiving a LT from a living donor (hazards ratio, 3.77; 95% confidence interval, 1.47-9.67; P = 0.006). Forty-one percent of CDI cases occurred within 1 week of LT. Seven percent of patients with CDI had a serum white blood cell count greater than 12 000 cells per μL, and 26% had a temperature greater than 38.0°C. After treatment 6 (22%) patients developed CDI relapse, and all were successfully treated. No patients died of CDI after a mean follow-up time of 1.8 years; however, overall survival was significantly lower among those with CDI (78% vs 92%; P = 0.033). Clostridium difficile infection after LT was associated with higher model for end-stage liver disease scores and receiving a LT from a living donor. Clostridium difficile infection often occurred soon after LT and was infrequently associated with leukocytosis or fever. Clostridium difficile infection in LT recipients was associated with lower overall survival.

  12. Prevalence of Clostridium Difficile Infection in Patients After Radical Cystectomy and Neoadjuvant Chemotherapy.

    Science.gov (United States)

    Cotter, Katherine J; Fan, Yunhua; Sieger, Gretchen K; Weight, Christopher J; Konety, Badrinath R

    2017-10-27

    Clostridium Difficile is the most common cause of nosocomial infectious diarrhea. This study evaluates the prevalence and predictors of Clostridium Difficile infections in patients undergoing radical cystectomy with or without neoadjuvant chemotherapy. Retrospective chart review was performed of all patients undergoing cystectomy and urinary diversion at a single institution from 2011-2017. Infection was documented in all cases with testing for Clostridium Difficile polymerase chain reaction toxin B. Patient and disease related factors were compared for those who received neoadjuvant chemotherapy vs. those who did not in order to identify potential risk factors associated with C. Difficile infections. Chi squared test and logistic regression analysis were used to determine statistical significance. Of 350 patients who underwent cystectomy, 41 (11.7%) developed Clostridium Difficile in the 30 day post-operative period. The prevalence of C. Difficile infection was higher amongst the patients undergoing cystectomy compared to the non-cystectomy admissions at our hospital (11.7 vs. 2.9%). Incidence was not significantly different among those who underwent cystectomy for bladder cancer versus those who underwent the procedure for other reasons. Median time to diagnosis was 6 days (range 3-28 days). The prevalence of C. Diff infections was not significantly different among those who received neoadjuvant chemotherapy vs. those who did not (11% vs. 10.4% p  = 0.72). A significant association between C. Difficile infection was not seen with proton pump inhibitor use ( p  = 0.48), patient BMI ( p  = 0.67), chemotherapeutic regimen ( p  = 0.94), individual surgeon ( p  = 0.54), type of urinary diversion (0.41), or peri-operative antibiotic redosing ( p  = 0.26). Clostridium Difficile infection has a higher prevalence in patients undergoing cystectomy. No significant association between prevalence and exposure to neoadjuvant chemotherapy was seen.

  13. Effect of tcdR Mutation on Sporulation in the Epidemic Clostridium difficile Strain R20291.

    Science.gov (United States)

    Girinathan, Brintha P; Monot, Marc; Boyle, Daniel; McAllister, Kathleen N; Sorg, Joseph A; Dupuy, Bruno; Govind, Revathi

    2017-01-01

    Clostridium difficile is an important nosocomial pathogen and the leading cause of hospital-acquired diarrhea. Antibiotic use is the primary risk factor for the development of C. difficile -associated disease because it disrupts normally protective gut flora and enables C. difficile to colonize the colon. C. difficile damages host tissue by secreting toxins and disseminates by forming spores. The toxin-encoding genes, tcdA and tcdB , are part of a pathogenicity locus, which also includes the tcdR gene that codes for TcdR, an alternate sigma factor that initiates transcription of tcdA and tcdB genes. We created a tcdR mutant in epidemic-type C. difficile strain R20291 in an attempt to identify the global role of tcdR . A site-directed mutation in tcdR affected both toxin production and sporulation in C. difficile R20291. Spores of the tcdR mutant were more heat sensitive than the wild type (WT). Nearly 3-fold more taurocholate was needed to germinate spores from the tcdR mutant than to germinate the spores prepared from the WT strain. Transmission electron microscopic analysis of the spores also revealed a weakly assembled exosporium on the tcdR mutant spores. Accordingly, comparative transcriptome analysis showed many differentially expressed sporulation genes in the tcdR mutant compared to the WT strain. These data suggest that regulatory networks of toxin production and sporulation in C. difficile strain R20291 a re linked with each other. IMPORTANCE C. difficile infects thousands of hospitalized patients every year, causing significant morbidity and mortality. C. difficile spores play a pivotal role in the transmission of the pathogen in the hospital environment. During infection, the spores germinate, and the vegetative bacterial cells produce toxins that damage host tissue. Thus, sporulation and toxin production are two important traits of C. difficile . In this study, we showed that a mutation in tcdR , the toxin gene regulator, affects both toxin

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

    Science.gov (United States)

    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 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 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.

  15. CodY-Dependent Regulation of Sporulation in Clostridium difficile.

    Science.gov (United States)

    Nawrocki, Kathryn L; Edwards, Adrianne N; Daou, Nadine; Bouillaut, Laurent; McBride, Shonna M

    2016-08-01

    Clostridium difficile must form a spore to survive outside the gastrointestinal tract. The factors that trigger sporulation in C. difficile remain poorly understood. Previous studies have suggested that a link exists between nutritional status and sporulation initiation in C. difficile In this study, we investigated the impact of the global nutritional regulator CodY on sporulation in C. difficile strains from the historical 012 ribotype and the current epidemic 027 ribotype. Sporulation frequencies were increased in both backgrounds, demonstrating that CodY represses sporulation in C. difficile The 027 codY mutant exhibited a greater increase in spore formation than the 012 codY mutant. To determine the role of CodY in the observed sporulation phenotypes, we examined several factors that are known to influence sporulation in C. difficile Using transcriptional reporter fusions and quantitative reverse transcription-PCR (qRT-PCR) analysis, we found that two loci associated with the initiation of sporulation, opp and sinR, are regulated by CodY. The data demonstrate that CodY is a repressor of sporulation in C. difficile and that the impact of CodY on sporulation and expression of specific genes is significantly influenced by the strain background. These results suggest that the variability of CodY-dependent regulation is an important contributor to virulence and sporulation in current epidemic isolates. This report provides further evidence that nutritional state, virulence, and sporulation are linked in C. difficile This study sought to examine the relationship between nutrition and sporulation in C. difficile by examining the global nutritional regulator CodY. CodY is a known virulence and nutritional regulator of C. difficile, but its role in sporulation was unknown. Here, we demonstrate that CodY is a negative regulator of sporulation in two different ribotypes of C. difficile We also demonstrate that CodY regulates known effectors of sporulation, Opp and Sin

  16. Growth of Clostridium perfringens during cooling of refried beans.

    Science.gov (United States)

    Cevallos-Cevallos, Juan M; Akins, E Deann; Friedrich, Loretta M; Danyluk, Michelle D; Simonne, Amarat H

    2012-10-01

    Outbreaks of Clostridium perfringens have been associated with dishes containing refried beans from food service establishments. However, growth of C. perfringens in refried beans has not been investigated, and predictive models have not been validated in this food matrix. We investigated the growth of C. perfringens during the cooling of refried beans. Refried beans (pinto and black, with and without salt added) were inoculated with 3 log CFU/g C. perfringens spores and incubated isothermally at 12, 23, 30, 35, 40, 45, and 50°C. The levels of C. perfringens were monitored 3, 5, 8, and 10 h after inoculation, and then fitted to the Baranyi primary model and the Rosso secondary model prior to solving the Baranyi differential equation. The final model was validated by dynamic cooling experiments carried out in stockpots, thus mimicking the worst possible food service conditions. All refried beans samples supported the growth of C. perfringens, and all models fit the data with pseudo-R(2) values of 0.95 or greater and mean square errors of 0.3 or lower. The estimated maximum specific growth rates were generally higher in pinto beans, with or without salt added (2.64 and 1.95 h(-1), respectively), when compared with black beans, with or without salt added (1.78 and 1.61 h(-1), respectively). After 10 h of incubation, maximum populations of C. perfringens were significantly higher in samples with no salt added (7.9 log CFU/g for both pinto and black beans) than in samples with salt added (7.3 and 7.2 log CFU/g for pinto and black beans, respectively). The dynamic model predicted the growth of C. perfringens during cooling, with an average root mean squared error of 0.44. The use of large stockpots to cool refried beans led to an observed 1.2-log increase (1.5-log increase predicted by model) in levels of C. perfringens during cooling. The use of shallower pans for cooling is recommended, because they cool faster, therefore limiting the growth of C. perfringens.

  17. The spore differentiation pathway in the enteric pathogen Clostridium difficile.

    Directory of Open Access Journals (Sweden)

    Fátima C Pereira

    Full Text Available Endosporulation is an ancient bacterial developmental program that culminates with the differentiation of a highly resistant endospore. In the model organism Bacillus subtilis, gene expression in the forespore and in the mother cell, the two cells that participate in endospore development, is governed by cell type-specific RNA polymerase sigma subunits. σ(F in the forespore, and σ(E in the mother cell control early stages of development and are replaced, at later stages, by σ(G and σ(K, respectively. Starting with σ(F, the activation of the sigma factors is sequential, requires the preceding factor, and involves cell-cell signaling pathways that operate at key morphological stages. Here, we have studied the function and regulation of the sporulation sigma factors in the intestinal pathogen Clostridium difficile, an obligate anaerobe in which the endospores are central to the infectious cycle. The morphological characterization of mutants for the sporulation sigma factors, in parallel with use of a fluorescence reporter for single cell analysis of gene expression, unraveled important deviations from the B. subtilis paradigm. While the main periods of activity of the sigma factors are conserved, we show that the activity of σ(E is partially independent of σ(F, that σ(G activity is not dependent on σ(E, and that the activity of σ(K does not require σ(G. We also show that σ(K is not strictly required for heat resistant spore formation. In all, our results indicate reduced temporal segregation between the activities of the early and late sigma factors, and reduced requirement for the σ(F-to-σ(E, σ(E-to-σ(G, and σ(G-to-σ(K cell-cell signaling pathways. Nevertheless, our results support the view that the top level of the endosporulation network is conserved in evolution, with the sigma factors acting as the key regulators of the pathway, established some 2.5 billion years ago upon its emergence at the base of the Firmicutes Phylum.

  18. Strategy to inactivate Clostridium perfringens spores in meat products.

    Science.gov (United States)

    Akhtar, Saeed; Paredes-Sabja, Daniel; Torres, J Antonio; Sarker, Mahfuzur R

    2009-05-01

    The current study aimed to develop an inactivation strategy for Clostridium perfringens spores in meat through a combination of spore activation at low pressure (100-200 MPa, 7 min) and elevated temperature (80 degrees C, 10 min); spore germination at high temperatures (55, 60 or 65 degrees C); and inactivation of germinated spores with elevated temperatures (80 and 90 degrees C, 10 and 20 min) and high pressure (586 MPa, at 23 and 73 degrees C, 10 min). Low pressures (100-200 MPa) were insufficient to efficiently activate C. perfringens spores for germination. However, C. perfringens spores were efficiently activated with elevated temperature (80 degrees C, 10 min), and germinated at temperatures lethal for vegetative cells (>or= 55 degrees C) when incubated for 60 min with a mixture of L-asparagine and KCl (AK) in phosphate buffer (pH 7) and in poultry meat. Inactivation of spores (approximately 4 decimal reduction) in meat by elevated temperatures (80-90 degrees C for 20 min) required a long germination period (55 degrees C for 60 min). However, similar inactivation level was reached with shorter germination period (55 degrees C for 15 min) when spore contaminated-meat was treated with pressure-assisted thermal processing (568 MPa, 73 degrees C, 10 min). Therefore, the most efficient strategy to inactivate C. perfringens spores in poultry meat containing 50 mM AK consisted: (i) a primary heat treatment (80 degrees C, 10 min) to pasteurize and denature the meat proteins and to activate C. perfringens spores for germination; (ii) cooling of the product to 55 degrees C in about 20 min and further incubation at 55 degrees C for about 15 min for spore germination; and (iii) inactivation of germinated spores by pressure-assisted thermal processing (586 MPa at 73 degrees C for 10 min). Collectively, this study demonstrates the feasibility of an alternative and novel strategy to inactivate C. perfringens spores in meat products formulated with germinants specific for C

  19. Burden of Clostridium difficile infection in the United States.

    Science.gov (United States)

    Lessa, Fernanda C; Mu, Yi; Bamberg, Wendy M; Beldavs, Zintars G; Dumyati, Ghinwa K; Dunn, John R; Farley, Monica M; Holzbauer, Stacy M; Meek, James I; Phipps, Erin C; Wilson, Lucy E; Winston, Lisa G; Cohen, Jessica A; Limbago, Brandi M; Fridkin, Scott K; Gerding, Dale N; McDonald, L Clifford

    2015-02-26

    The magnitude and scope of Clostridium difficile infection in the United States continue to evolve. In 2011, we performed active population- and laboratory-based surveillance across 10 geographic areas in the United States to identify cases of C. difficile infection (stool specimens positive for C. difficile on either toxin or molecular assay in residents ≥ 1 year of age). Cases were classified as community-associated or health care-associated. In a sample of cases of C. difficile infection, specimens were cultured and isolates underwent molecular typing. We used regression models to calculate estimates of national incidence and total number of infections, first recurrences, and deaths within 30 days after the diagnosis of C. difficile infection. A total of 15,461 cases of C. difficile infection were identified in the 10 geographic areas; 65.8% were health care-associated, but only 24.2% had onset during hospitalization. After adjustment for predictors of disease incidence, the estimated number of incident C. difficile infections in the United States was 453,000 (95% confidence interval [CI], 397,100 to 508,500). The incidence was estimated to be higher among females (rate ratio, 1.26; 95% CI, 1.25 to 1.27), whites (rate ratio, 1.72; 95% CI, 1.56 to 2.0), and persons 65 years of age or older (rate ratio, 8.65; 95% CI, 8.16 to 9.31). The estimated number of first recurrences of C. difficile infection was 83,000 (95% CI, 57,000 to 108,900), and the estimated number of deaths was 29,300 (95% CI, 16,500 to 42,100). The North American pulsed-field gel electrophoresis type 1 (NAP1) strain was more prevalent among health care-associated infections than among community-associated infections (30.7% vs. 18.8%, Pdifficile was responsible for almost half a million infections and was associated with approximately 29,000 deaths in 2011. (Funded by the Centers for Disease Control and Prevention.).

  20. Radioprotective preparation

    International Nuclear Information System (INIS)

    Stefanova, D.; Frattadochi, A.; Gattavecchia, E.; Ferri, E.; Tonnelli, D.

    1988-01-01

    The invention is intended for radiation injuries prophylaxis in mammals. It has an well expressed radioprotective effect against acute gamma irradiation on cellular level as well as a prolonged action when applied up to 48 hours before the acute irradiation. The preparation is a coprecipitate of the natural tripeptide glutathione (reduced form) and polyvinyl pyrrolidone (pvp) in ratio 30-60/70-40. It is obtained by incubation method with subsequent lyophilization from water solution of the initial components. The molecular mass of the pvp is 20 till 360.10 3 . 2 claims

  1. Target preparation

    International Nuclear Information System (INIS)

    Hinn, G.M.

    1984-01-01

    A few of the more interesting of the 210 targets prepared in the Laboratory last year are listed. In addition the author continues to use powdered silver mixed with /sup 9,10/BeO to produce sources for accelerator radio dating of Alaskan and South Polar snow. Currently, he is trying to increase production by multiple sample processing. Also the author routinely makes 3 μg/cm 2 cracked slacked carbon stripper foils and is continuing research with some degree of success in making enriched 28 Si targets starting with the oxide

  2. Effectiveness of various cleaning and disinfectant products on Clostridium difficile spores of PCR ribotypes 010, 014 and 027

    NARCIS (Netherlands)

    Kenters, N.; Huijskens, E.G.; Wit, S.C.J. de; Sanders, I.; Rosmalen, J. van; Kuijper, E.J.; Voss, A.

    2017-01-01

    BACKGROUND: In healthcare facilities, Clostridium difficile infections spread by transmission of bacterial spores. Appropriate sporicidal disinfectants are needed to prevent development of clusters and outbreaks. In this study different cleaning/disinfecting wipes and sprays were tested for their

  3. Effectiveness of various cleaning and disinfectant products on Clostridium difficile spores of PCR ribotypes 010, 014 and 027

    NARCIS (Netherlands)

    Kenters, N.; E. Huijskens (Elisabeth); de Wit, S.C.J.; Sanders, I.G.J.M.; J.M. van Rosmalen (Joost); E. Kuijper; Voss, A.

    2017-01-01

    textabstractBackground: In healthcare facilities, Clostridium difficile infections spread by transmission of bacterial spores. Appropriate sporicidal disinfectants are needed to prevent development of clusters and outbreaks. In this study different cleaning/disinfecting wipes and sprays were tested

  4. The Role of Clostridium Perfringens in the Syndrome of Intestinal Affection in Children and Possibilities of Drug Correction

    Directory of Open Access Journals (Sweden)

    G.O. Lezhenko

    2014-09-01

    Full Text Available The article showed the features of clostridiosis course, caused by Clostridium perfringens, in children of different age groups taking into account the dose of pathogen in feces and pathogenetically grounded possibilities of etiological therapy.

  5. Management of Clostridium difficile diarrhoea in District General Hospital: audit for 3months and review of literature

    OpenAIRE

    Dr. J. Sellers; Dr. Varun Dixit

    2007-01-01

    Clostridium difficile is one of the most important causes of diarrhoea especially following antibiotic course. Elderly population is more susceptible and results in significant mortality and morbidity. We audited twenty four cases of Clostridium difficile in our hospital over duration of three months. We looked into the demographic features of the patient population and compliance with the Trust guidelines for the management of the diarrhoea.

  6. Management of Clostridium difficile diarrhoea in District General Hospital: audit for 3months and review of literature

    Directory of Open Access Journals (Sweden)

    Dr. J. Sellers

    2007-01-01

    Full Text Available Clostridium difficile is one of the most important causes of diarrhoea especially following antibiotic course. Elderly population is more susceptible and results in significant mortality and morbidity. We audited twenty four cases of Clostridium difficile in our hospital over duration of three months. We looked into the demographic features of the patient population and compliance with the Trust guidelines for the management of the diarrhoea.

  7. Cannabidiol restores intestinal barrier dysfunction and inhibits the apoptotic process induced by Clostridium difficile toxin A in Caco-2 cells.

    Science.gov (United States)

    Gigli, Stefano; Seguella, Luisa; Pesce, Marcella; Bruzzese, Eugenia; D'Alessandro, Alessandra; Cuomo, Rosario; Steardo, Luca; Sarnelli, Giovanni; Esposito, Giuseppe

    2017-12-01

    Clostridium difficile toxin A is responsible for colonic damage observed in infected patients. Drugs able to restore Clostridium difficile toxin A-induced toxicity have the potential to improve the recovery of infected patients. Cannabidiol is a non-psychotropic component of Cannabis sativa, which has been demonstrated to protect enterocytes against chemical and/or inflammatory damage and to restore intestinal mucosa integrity. The purpose of this study was to evaluate (a) the anti-apoptotic effect and (b) the mechanisms by which cannabidiol protects mucosal integrity in Caco-2 cells exposed to Clostridium difficile toxin A. Caco-2 cells were exposed to Clostridium difficile toxin A (30 ng/ml), with or without cannabidiol (10 -7 -10 -9  M), in the presence of the specific antagonist AM251 (10 -7  M). Cytotoxicity assay, transepithelial electrical resistence measurements, immunofluorescence analysis and immunoblot analysis were performed in the different experimental conditions. Clostridium difficile toxin A significantly decreased Caco-2 cells' viability and reduced transepithelial electrical resistence values and RhoA guanosine triphosphate (GTP), bax, zonula occludens-1 and occludin protein expression, respectively. All these effects were significantly and concentration-dependently inhibited by cannabidiol, whose effects were completely abolished in the presence of the cannabinoid receptor type 1 (CB1) antagonist, AM251. Cannabidiol improved Clostridium difficile toxin A-induced damage in Caco-2 cells, by inhibiting the apoptotic process and restoring the intestinal barrier integrity, through the involvement of the CB1 receptor.

  8. MALDI-TOF MS is more accurate than VITEK II ANC card and API Rapid ID 32 A system for the identification of Clostridium species.

    Science.gov (United States)

    Kim, Young Jin; Kim, Si Hyun; Park, Hyun-Jung; Park, Hae-Geun; Park, Dongchul; Song, Sae Am; Lee, Hee Joo; Yong, Dongeun; Choi, Jun Yong; Kook, Joong-Ki; Kim, Hye Ran; Shin, Jeong Hwan

    2016-08-01

    All 50 Clostridium difficile strains were definitely identified by Vitek2 system, Rapid ID 32A system, and MALDI-TOF. For 18 non-difficile Clostridium strains, the identification results were correct in 0, 2, and 17 strains by Vitek2, Rapid ID 32A, and MALDI-TOF, respectively. MALDI-TOF could be used as the primary tool for identification of Clostridium species. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. The Structure of the Neurotoxin- Associated Protein HA33/A from Clostridium botulinum Suggests a Reoccurring Beta-Trefoil Fold in the Progenitor Toxin Complex

    National Research Council Canada - National Science Library

    Arndt, Joseph W; Gu, Jenny; Jaroszewski, Lukasz; Schwarzenbacher, Robert; Hanson, Michael A; Lebeda, Frank L; Stevens, Raymond C

    2004-01-01

    The hemagglutinating protein HA33 from Clostridium botulinum is associated with the large botulinum neurotoxin secreted complexes and is critical in toxin protection, internalization, and possibly activation...

  10. Initiation of sporulation in Clostridium difficile: a twist on the classic model.

    Science.gov (United States)

    Edwards, Adrianne N; McBride, Shonna M

    2014-09-01

    The formation of dormant endospores is a complex morphological process that permits long-term survival in inhospitable environments for many Gram-positive bacteria. Sporulation for the anaerobic gastrointestinal pathogen Clostridium difficile is necessary for survival outside of the gastrointestinal tract of its host. While the developmental stages of spore formation are largely conserved among endospore-forming bacteria, the genus Clostridium appears to be missing a number of conserved regulators required for efficient sporulation in other spore-forming bacteria. Several recent studies have discovered novel mechanisms and distinct regulatory pathways that control the initiation of sporulation and early-sporulation-specific gene expression. These differences in regulating the decision to undergo sporulation reflects the unique ecological niche and environmental conditions that C. difficile inhabits and encounters within the mammalian host. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  11. Imipenem-induced clostridium difficile diarrhea in a patient with chronic renal failure

    Directory of Open Access Journals (Sweden)

    R Enríquez

    2011-01-01

    Full Text Available An 80-year-old man was diagnosed to have pneumonia and advanced chronic kidney disease. He presented with anuria and hemodialysis, by temporary femoral catheter, was initiated. He was empirically treated with imipenem/cilastatin 500 mg/24 h after hemodialysis. After 10 days of antibiotic intake, he developed severe diarrhea. Diagnosis of Clostridium difficile (CD-associated diarrhea was confirmed by detection of the toxins A and B in his stool. Imipenem therapy was discontinued; Vancomycin 500 mg orally every 6 h and 1000 mg per rectum every day was added. After two weeks of this treatment, the patient reported complete resolution of the diarrhea and stool samples were negative for Clostridium toxin. In this case, the most possible cause of CD colitis was considered to be imipenem because of the temporal relationship between exposure to the drug and onset of symptoms.

  12. The incidence of Clostridioides difficile and Clostridium perfringens netF-positive strains in diarrheic dogs.

    Science.gov (United States)

    Diniz, Amanda Nadia; Coura, Fernanda Morcatti; Rupnik, Maja; Adams, Vicki; Stent, Thomas L; Rood, Julian I; de Oliveira, Carlos Augusto; Lobato, Francisco Carlos Faria; Silva, Rodrigo Otávio Silveira

    2018-02-01

    The aim of this study was to examine the incidence of Clostridioides (previously Clostridium) difficile and Clostridium perfringens in the feces of diarrheic and non-diarrheic dogs. Also, the presence of other common canine enteropathogens was examined. Toxigenic C. difficile and C. perfringens positive for the NetF-encoding gene (netF) were detected in 11 (11.9%) and seven (7.6%) diarrheic dogs, respectively. Three dogs were diagnosed simultaneously with toxigenic C. difficile and netF-positive C. perfringens. Among other enteropathogens, Giardia sp. was the most common agent detected in dogs positive for toxigenic C. difficile or netF-positive C. perfringens. The results suggest that C. difficile and C. perfringens occur more frequently as a primary cause of diarrhea. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Clostridium perfringens and C. difficile in parvovirus-positive dogs.

    Science.gov (United States)

    Silva, Rodrigo Otávio Silveira; Dorella, Fernanda Alves; Figueiredo, Henrique Cesar Pereira; Costa, Érica Azevedo; Pelicia, Vanessa; Ribeiro, Bruna Letícia Devidé; Ribeiro, Marcio Garcia; Paes, Antonio Carlos; Megid, Jane; Lobato, Francisco Carlos Faria

    2017-12-01

    The aim of this study was to investigate Clostridium difficile and Clostridium perfringens in 82 diarrheic dogs positive for canine parvovirus type 2 (CPV). Enterotoxigenic C. perfringens type A was isolated from three (3.6%) dogs. One (1.2%) strain was also positive for NetE- and NetF-encoding genes, which are commonly associated with diarrhea in dogs. Toxigenic C. difficile was isolated from one animal (1.2%), which was also positive for A/B toxins. The present study identified C. difficile and C. perfringens infection in CPV-positive dogs. Further studies are necessary to clarify if clostridial infections may predispose or potentiate CPV-infection in dogs or vice versa. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Cecal Perforation Associated with Clostridium difficile Infection: A Case Report.

    Science.gov (United States)

    Luthe, Sarah Kyuragi; Sato, Ryota

    2017-04-01

    Various complications are reported with Clostridium difficile infection (CDI), including fulminant CDI. Fulminant CDI is an underappreciated life-threatening condition associated with complications such as toxic megacolon and bowel perforation. A 79-year-old woman presented to the Emergency Department with altered mental status. She was admitted and conservatively treated for a left thalamic hemorrhage. While hospitalized, she developed watery diarrhea due to Clostridium difficile. Although metronidazole was initiated, she developed altered mental status and septic shock. Abdominal x-ray study and computed tomography revealed a significantly dilatated colon and a massive pneumoperitoneum. She underwent subtotal colectomy with a 14-day course of intravenous meropenem. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: This case suggests that we must be aware of the complications that CDI may present and adequately consider surgical management because early diagnosis and surgical treatment is critical to reduce the mortality of fulminant CDI. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Clostridium difficile Recombinant Toxin A Repeating Units as a Carrier Protein for Conjugate Vaccines: Studies of Pneumococcal Type 14, Escherichia coli K1, and Shigella flexneri Type 2a Polysaccharides in Mice

    Science.gov (United States)

    Pavliakova, Danka; Moncrief, J. Scott; Lyerly, David M.; Schiffman, Gerald; Bryla, Dolores A.; Robbins, John B.; Schneerson, Rachel

    2000-01-01

    Unlike the native protein, a nontoxic peptide (repeating unit of the native toxin designated rARU) from Clostridium difficile toxin A (CDTA) afforded an antigen that could be bound covalently to the surface polysaccharides of pneumococcus type 14, Shigella flexneri type 2a, and Escherichia coli K1. The yields of these polysaccharide-protein conjugates were significantly increased by prior treatment of rARU with succinic anhydride. Conjugates, prepared with rARU or succinylated (rARUsucc), were administered to mice by a clinically relevant dosage and immunization scheme. All conjugates elicited high levels of serum immunoglobulin G both to the polysaccharides and to CDTA. Conjugate-induced anti-CDTA had neutralizing activity in vitro and protected mice challenged with CDTA, similar to the rARU alone. Conjugates prepared with succinylated rARU, therefore, have potential for serving both as effective carrier proteins for polysaccharides and for preventing enteric disease caused by C. difficile. PMID:10722615

  16. Clostridium perfringens types A and D associated with enterotoxemia in an 18-month-old goat

    Directory of Open Access Journals (Sweden)

    S. Miyashiro

    2007-01-01

    Full Text Available Postmortem examination of a Boer buck that died peracutely revealed bowel and liver diffusely congested and edematous. Kidney was apparently edematous. Clostridium perfringens type A was isolated from bowel and type D from kidney. Microscopic examination revealed large areas of necrosis in the renal cortex and medulla (pulpy kidney disease, hyperemia and centrilobular necrosis of the liver, necrosis of the small-intestine wall, pulmonary edema and congestion, intense hyperemia of the cerebellum, hyperemia and edema of the brain.

  17. Isolation of Clostridium difficile and Detection of A and B Toxins Encoding Genes

    OpenAIRE

    Abbas Ali Imani Fooladi; Sadegh Rahmati; Jalil Falah Mehr Abadi; Raheleh Halabian; Hamid Sedighian; Mohammad Javad Soltanpour; Mohsen Rahimi

    2014-01-01

    Background: Clostridium difficile is the most important anaerobic, gram positive, spore forming bacillus which is known as a prevalent factor leading to antibiotic associated diarrheas and is the causative agent of pseudomembrane colitis. The role of this bacterium along with the over use of antibiotics have been proved to result in colitis. The major virulence factors of these bacteria are the A and B toxins. Objectives: The purpose of this study was to isolate C. difficile from sto...

  18. Modeling of Clostridium tyrobutyricum for Butyric Acid Selectivity in Continuous Fermentation

    OpenAIRE

    Du, Jianjun; McGraw, Amy; Hestekin, Jamie

    2014-01-01

    A mathematical model was developed to describe batch and continuous fermentation of glucose to organic acids with Clostridium tyrobutyricum. A modified Monod equation was used to describe cell growth, and a Luedeking-Piret equation was used to describe the production of butyric and acetic acids. Using the batch fermentation equations, models predicting butyric acid selectivity for continuous fermentation were also developed. The model showed that butyric acid production was a strong function ...

  19. Modeling of Clostridium t yrobutyricum for Butyric Acid Selectivity in Continuous Fermentation

    OpenAIRE

    Jianjun Du; Amy McGraw; Jamie A. Hestekin

    2014-01-01

    A mathematical model was developed to describe batch and continuous fermentation of glucose to organic acids with Clostridium tyrobutyricum . A modified Monod equation was used to describe cell growth, and a Luedeking-Piret equation was used to describe the production of butyric and acetic acids. Using the batch fermentation equations, models predicting butyric acid selectivity for continuous fermentation were also developed. The model showed that butyric acid production was a strong function...

  20. Bacteraemia and breast abscess: unusual extra-intestinal manifestations of Clostridium difficile infection.

    Science.gov (United States)

    Durojaiye, Oyewole; Gaur, Soma; Alsaffar, Layth

    2011-03-01

    Extra-intestinal manifestations of Clostridium difficile infection are uncommon. Most cases are associated with gastrointestinal disease and often occur as a mixed infection with other gut flora. We report a case of breast abscess following monomicrobial C. difficile bacteraemia in a female with background chronic hepatitis C infection and alcoholic liver disease. No evidence of colitis was found. Our case shows that C. difficile is indeed capable of spreading from the gastrointestinal tract.

  1. Clostridium perfringens in London, July 2009: two weddings and an outbreak.

    OpenAIRE

    Eriksen, J.; Zenner, D.; Anderson, S. R.; Grant, K.; Kumar, D.

    2010-01-01

    : Food poisoning outbreaks caused by Clostridium perfringens enterotoxin occur occasionally in Europe but have become less common in recent years. This paper presents the microbiological and epidemiological results of a large C. perfringens outbreak occurring simultaneously at two weddings that used the same caterer. The outbreak involved several London locations and required coordination across multiple agencies. A case-control study (n=134) was carried out to analyse possible associations b...

  2. Clostridium beijerinckii mutant obtained atmospheric pressure glow discharge generates enhanced electricity in a microbial fuel cell.

    Science.gov (United States)

    Liu, Jun; Guo, Ting; Wang, Dong; Ying, Hanjie

    2015-01-01

    A Clostridium beijerinckii mutant M13 was derived from C. beijerinckii NCIMB 8052 by atmospheric pressure glow discharge. C. beijerinckii M13 generated a maximum output power density of 79.2 mW m(-2) and a maximum output voltage of 230 mV in a microbial fuel cell containing 1 g glucose l(-1) as carbon source and 0.15 g methyl viologen l(-1) as an electron carrier.

  3. Enhancing nitrite inhibition of Clostridium botulinum with isoascorbate in perishable canned cured meat.

    Science.gov (United States)

    Tompkin, R B; Christiansen, L N; Shaparis, A B

    1978-01-01

    Addition of sodium isoascorbate to the formulation for perishable canned comminuted cured meat markedly enhanced the efficacy of nitrite against Clostridium botulinum. This effect was reproducible through a series of three tests. In one test it was found that the initial addition of 50 microgram of sodium nitrite per g plus isoascorbate was as effective as 156 microgram of sodium nitrite per g alone. PMID:341810

  4. Necrotizing Fasciitis and Toxic Shock Syndrome from Clostridium septicum following a Term Cesarean Delivery

    Directory of Open Access Journals (Sweden)

    B. H. Rimawi

    2014-01-01

    Full Text Available Necrotizing fasciitis and toxic shock syndrome are life-threatening conditions that can be seen after any surgical procedure. With only 4 previous published case reports in the obstetrics and gynecology literature of these two conditions occurring secondary to Clostridium septicum, we describe a case of necrotizing fasciitis and toxic shock syndrome occurring after a term cesarean delivery caused by this microorganism, requiring aggressive medical and surgical intervention.

  5. Administration of probiotic kefir to mice with Clostridium difficile infection exacerbates disease.

    Science.gov (United States)

    Spinler, Jennifer K; Brown, Aaron; Ross, Caná L; Boonma, Prapaporn; Conner, Margaret E; Savidge, Tor C

    2016-08-01

    Lifeway(®) kefir, a fermented milk product containing 12 probiotic organisms, is reported to show promise as an alternative to fecal microbiota transplantation for recurrent Clostridium difficile infection (CDI). We employed a murine CDI model to study the probiotic protective mechanisms and unexpectedly determined that kefir drastically increased disease severity. Our results emphasize the need for further independent clinical testing of kefir as alternative therapy in recurrent CDI. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Impact of end stage kidney disease on costs and outcomes of Clostridium difficile infection

    OpenAIRE

    Abhinav Goyal; Kshitij Chatterjee; Sujani Yadlapati; Janani Rangaswami

    2017-01-01

    Objectives: To assess the impact of end stage kidney disease (ESKD) on the outcomes of Clostridium difficile infection (CDI), including complications of infection, length of hospital stay, overall mortality, and healthcare burden. Methods: The National Inpatient Sample (NIS) database created by the Agency of Healthcare Research and Quality (AHRQ) was used, covering the years 2009 through 2013. Manufacturer-provided sampling weights were used to produce national estimates. Results: All-c...

  7. Prevalence and Genotypic Characteristics of Clostridium difficile in a Closed and Integrated Human and Swine Population▿

    OpenAIRE

    Norman, Keri N.; Scott, H. Morgan; Harvey, Roger B.; Norby, Bo; Hume, Michael E.; Andrews, Kathleen

    2011-01-01

    Recently, an apparent rise in the number of cases attributed to community-acquired Clostridium difficile infection has led researchers to explore additional sources of infection. The finding of C. difficile in food animals and retail meat has raised concern about potential food-borne and occupational exposures. The objective of this study was to compare C. difficile isolated from a closed population of healthy individuals consisting of both humans and swine in order to investigate possible fo...

  8. Clostridium perfringens Antigens Recognized by Broiler Chickens Immune to Necrotic Enteritis▿

    OpenAIRE

    Kulkarni, R. R.; Parreira, V. R.; Sharif, S.; Prescott, J. F.

    2006-01-01

    Little is known about immunity to necrotic enteritis (NE) in chickens. A recent study of broiler chickens showed that protection against NE was associated with infection-immunization with virulent but not with avirulent Clostridium perfringens.In the current study, six secreted antigenic proteins unique to virulent C. perfringens that reacted to serum antibodies from immune birds were identified by mass spectrophotometry; three of these proteins are part of the VirR-VirS regulon.

  9. [A case of freeze-dried gas gangrene antitoxin for the treatment of Clostridium perfringens sepsis].

    Science.gov (United States)

    Yoshida, Juichiro; Nakamura, Hideki; Yamada, Shinya; Sekoguchi, Satoru; Suzuki, Takahiro; Tomatsuri, Naoya; Sato, Hideki; Okuyama, Yusuke; Kimura, Hiroyuki; Yoshida, Norimasa

    2015-02-01

    A 66-year-old man was admitted to our hospital with high fever. We diagnosed a gas-containing liver abscess and performed percutaneous abscess drainage. However, 15 hours after admission, he developed massive intravascular hemolysis and acidosis. Sepsis due to Clostridium perfringens was suspected and we treated the patient intensively with multidisciplinary approaches, including antibiotics, mechanical ventilation, and renal replacement therapy. Furthermore, we administered freeze-dried gas gangrene antitoxin. Despite intensive care, the patient died 43 hours after admission.

  10. Transcriptomic Analysis of (Group I) Clostridium botulinum ATCC 3502 Cold Shock Response

    OpenAIRE

    Dahlsten, Elias; Isokallio, Marita; Somervuo, Panu; Lindström, Miia; Korkeala, Hannu

    2014-01-01

    Profound understanding of the mechanisms foodborne pathogenic bacteria utilize in adaptation to the environmental stress they encounter during food processing and storage is of paramount importance in design of control measures. Chill temperature is a central control measure applied in minimally processed foods; however, data on the mechanisms the foodborne pathogen Clostridium botulinum activates upon cold stress are scarce. Transcriptomic analysis on the C. botulinum ATCC 3502 strain upon t...

  11. Saccharomyces boulardii Stimulates Intestinal Immunoglobulin A Immune Response to Clostridium difficile Toxin A in Mice

    Science.gov (United States)

    Qamar, Amir; Aboudola, Samer; Warny, Michel; Michetti, Pierre; Pothoulakis, Charalabos; LaMont, J. Thomas; Kelly, Ciarán P.

    2001-01-01

    Saccharomyces boulardii is a nonpathogenic yeast that protects against antibiotic-associated diarrhea and recurrent Clostridium difficile colitis. The administration of C. difficile toxoid A by gavage to S. boulardii-fed BALB/c mice caused a 1.8-fold increase in total small intestinal immunoglobulin A levels (P = 0.003) and a 4.4-fold increase in specific intestinal anti-toxin A levels (P boulardii-mediated protection against diarrheal illnesses. PMID:11254650

  12. Routine disc diffusion antimicrobial susceptibility testing of Clostridium difficile and association with PCR ribotype 027

    DEFF Research Database (Denmark)

    Holt, H M; Danielsen, T K; Justesen, U S

    2015-01-01

    Reduced susceptibility to metronidazole and vancomycin in Clostridium difficile has been reported, which emphasises the need for simple antimicrobial susceptibility testing methods. The aim of this study was to apply a published disc diffusion method and zone diameter breakpoint correlates...... the published breakpoint (difficile PCR ribotype 027 isolates had smaller zone...... diameters than non-027 isolates. The disc diffusion method is very simple and inexpensive, and the published zone diameter breakpoints will detect C. difficile isolates with reduced susceptibility to metronidazole and vancomycin....

  13. The Epidemiology and Economic Burden of Clostridium difficile Infection in Korea

    OpenAIRE

    Choi, Hyung-Yun; Park, So-Youn; Kim, Young-Ae; Yoon, Tai-Young; Choi, Joong-Myung; Choe, Bong-Keun; Ahn, So-Hee; Yoon, Seok-Jun; Lee, Ye-Rin; Oh, In-Hwan

    2015-01-01

    The prevalence of Clostridium difficile infection and the associated burden have recently increased in many countries. While the main risk factors for C. difficile infection include old age and antibiotic use, the prevalence of this infection is increasing in low-risk groups. These trends highlight the need for research on C. difficile infection. This study pointed out the prevalence and economic burden of C. difficile infection and uses the representative national data which is primarily fro...

  14. Vitamin D deficiency: A potential risk factor for Clostridium difficile infection

    OpenAIRE

    Youssef, Dima; Grant, William B; Peiris, Alan N

    2012-01-01

    Dima Youssef,1 William B Grant,2 Alan N Peiris3,41Department of Internal Medicine, Division of Infectious Diseases, 2Sunlight, Nutrition and Health Research Center, San Francisco, CA USA; 3Department of Medicine, Mountain Home VAMC, 4Department of Medicine, East Tennessee State University, Johnson City, Tennessee, USAIn the July 3, 2012 issue of the journal of Risk Management and Healthcare Policy, Martinez et al present a nice review on Clostridium difficile (C. difficile) infections.1 The d...

  15. THE POWER OF POOP: FECAL MICROBIOTA TRANSPLANTATION FOR CLOSTRIDIUM DIFFICILE INFECTION

    OpenAIRE

    EDMOND, MICHAEL B.

    2016-01-01

    The human gut is colonized with 200 to 1,000 bacterial species. Administration of antibiotics reduces the diversity of the intestinal microbiota, reduces colonization resistance, and can lead to infection with Clostridium difficile. These infections have become more prevalent and increasingly patients are experiencing multiple recurrences that are incurable with standard treatment. Although fecal microbiota transplantation (FMT) has been used for centuries in human and veterinary medicine, on...

  16. Coxofemoral luxation in a border collie as a complication of a Clostridium tetani infection.

    Science.gov (United States)

    Goldhammer, M A; Chapman, P S; Grierson, J M

    2008-03-01

    A four-month-old male, entire, border collie was presented to the Queen Mother Hospital for Animals with a two day history of muscular spasms and "Risus sardonicus". Tetanus was diagnosed, and the dog was treated with tetanus antitoxin, antibiotics and supportive therapy. Coxofemoral luxation resulted as a complication of the tetanus and was successfully managed by performing a femoral head and neck excision. This is the first report of joint luxation associated with Clostridium tetani infection in a dog.

  17. Anaerobic thermophilic culture-system

    Energy Technology Data Exchange (ETDEWEB)

    Ljungdahl, L G; Wiegel, J K.W.

    1981-04-14

    A mixed culture system of Thermoanaerobacter ethanolicus and Clostridium thermocellum is employed for anaerobic, thermophilic ethanol fermentation of cellulose. By cellulase action, monosaccharides are formed which inhibit the growth of C. thermocellum, but are fermented by T. ethanolicus. Thus, at a regulated pH-value of 7.5, this mixed culture system of micro organisms results in a cellulose fermentation with a considerably higher ethanol yield.

  18. Cloning and sequence analysis of hyaluronoglucosaminidase (nagH gene of Clostridium chauvoei

    Directory of Open Access Journals (Sweden)

    Saroj K. Dangi

    2017-09-01

    Full Text Available Aim: Blackleg disease is caused by Clostridium chauvoei in ruminants. Although virulence factors such as C. chauvoei toxin A, sialidase, and flagellin are well characterized, hyaluronidases of C. chauvoei are not characterized. The present study was aimed at cloning and sequence analysis of hyaluronoglucosaminidase (nagH gene of C. chauvoei. Materials and Methods: C. chauvoei strain ATCC 10092 was grown in ATCC 2107 media and confirmed by polymerase chain reaction (PCR using the primers specific for 16-23S rDNA spacer region. nagH gene of C. chauvoei was amplified and cloned into pRham-SUMO vector and transformed into Escherichia cloni 10G cells. The construct was then transformed into E. cloni cells. Colony PCR was carried out to screen the colonies followed by sequencing of nagH gene in the construct. Results: PCR amplification yielded nagH gene of 1143 bp product, which was cloned in prokaryotic expression system. Colony PCR, as well as sequencing of nagH gene, confirmed the presence of insert. Sequence was then subjected to BLAST analysis of NCBI, which confirmed that the sequence was indeed of nagH gene of C. chauvoei. Phylogenetic analysis of the sequence showed that it is closely related to Clostridium perfringens and Clostridium paraputrificum. Conclusion: The gene for virulence factor nagH was cloned into a prokaryotic expression vector and confirmed by sequencing.

  19. Complete genome sequence of Clostridium estertheticum DSM 8809, a microbe identified in spoiled vacuum packed beef

    Directory of Open Access Journals (Sweden)

    Zhongyi Yu

    2016-11-01

    Full Text Available Blown pack spoilage (BPS is a major issue for the beef industry. Aetiological agents of BPS involve members of a group of Clostridium species, including Clostridium estertheticum which has the ability to produce gas, mostly carbon dioxide, under anaerobic psychotrophic growth conditions. This spore-forming bacterium grows slowly under laboratory conditions, and it can take up to 3 months to produce a workable culture. These characteristics have limited the study of this commercially challenging bacterium. Consequently information on this bacterium is limited and no effective controls are currently available to confidently detect and manage this production risk. In this study the complete genome of Clostridium estertheticum DSM 8809 was determined by SMRT® sequencing. The genome consists of a circular chromosome of 4.7 Mbp along with a single plasmid carrying a potential tellurite resistance gene tehB and a Tn3-like resolvase-encoding gene tnpR. The genome sequence was searched for central metabolic pathways that would support its biochemical profile and several enzymes contributing to this phenotype were identified. Several putative antibiotic/biocide/metal resistance-encoding genes and virulence factors were also identified in the genome, a feature that requires further research. The availability of the genome sequence will provide a basic blueprint from which to develop valuable biomarkers that could support and improve the detection and control of this bacterium along the beef production chain.

  20. Pemanfaatan Serbuk Gergaji Menjadi Biobutanol dengan Hidrolisis Selulase dan Fermentasi Bakteri Clostridium Acetobutylicum

    Directory of Open Access Journals (Sweden)

    Hayuni Devina Fajariah

    2014-09-01

    Full Text Available Biobutanol adalah jenis alkohol ikatan C-4 (C4H9OH yang terbuat dari biomassa. Penelitian ini dilakukan dengan memanfaatkan limbah kayu yang dihasilkan dari proses penggergajian kayu yang mengandung selulosa (55%, hemiselulosa (14%, dan lignin (21%. Biobutanol diproduksi dengan cara hidrolisis enzim selulase dan fermentasi bakteri Clostridium acetobutylicum. Variabel pada penelitian ini adalah penambahan enzim selulase pada proses hidrolisis (penambahan enzim atau tanpa penambahan enzim, pH awal proses fermentasi (5 atau 7 dan jumlah penambahan starter bakteri Clostridium acetobutylicum (5 atau 10 ml dengan variasi lama proses fermentasi 2,4,6,8,10,12 hari. Parameter dalam penelitian ini adalah analisa kadar selulosa, gula tereduksi, dan kadar butanol. Berdasarkan hasil penetian, diketahui bahwa proses hidrolisis dengan penambahan enzim selulase, kondisi awal fermentasi pH 5 dan penambahan inokulum bakteri Clostridium acetobutylicum sebanyak 10 ml dengan lama waktu fermentasi 12 hari merupakan kondisi yang paling efektif menghasilkan kadar butanol tertinggi dari 50 gram limbah serbuk gergaji. Kadar butanol tertinggi sebesar 1,88 % dari 1 µL sampel hasil fermentasi yang diinjeksikan ke dalam kromatografi gas.

  1. DEVELOPMENT OF ENZYME-LINKAGE IMMUNOSORBENT ASSAY AGAINST TYPE B OF CLOSTRIDIUM BOTULINUM: A PRELIMINARY STUDY

    Directory of Open Access Journals (Sweden)

    S. N. Depamede

    2011-12-01

    Full Text Available Clostridium botulinum neurotoxin (BoNTs is one of the causes of economic loss in the livestock industry. This economic loss would be as a direct result when animals poisoned by BoNTs or indirectly when the livestock products are contaminated by BoNTs, which end up with the products are banned by authority. Therefore a routine surveillance of BoNTs in the farm and in livestock product processing industry is urgently needed. One of the most relatively quick and accurate methods to perform a routine detection of the presence of BoNTs is enzyme-linkage immunosorbant assay (ELISA. In this article we describe the results of the development of ELISA, using polyclonal antibodies against BoNTs-B produced locally. Antibodies were generated from six Balb/c mice with standard immunological methods. Mice were immunized three times for a period of 8 weeks with a commercial type B Clostridium botulinum toxoid at a dose of 100 ng per mouse per injection. The resulting antibody was purified by a combination of ammonium sulfate precipitation 50% (w/v technique and a protein A column method. The results of this preliminary study indicated that the developed ELISA method capable of detecting type B Clostridium botulinum toxin up to 1.0 ng/ml.

  2. Bio-assisted synthesis and characterization of nanostructured bismuth (III) sulphide using Clostridium acetobutylicum

    International Nuclear Information System (INIS)

    Kamaraj, Sathish Kumar; Venkatachalam, Ganesh; Arumugam, Palaniappan; Berchmans, Sheela

    2014-01-01

    Nanostructured bismuth (III) sulphide is synthesized at room temperature using a hydrogen sulphide producing microorganism namely Clostridium acetobutylicum. On contrary to chemical routes involving both the high and room temperature methods, the present experimental procedure involves a bio-assisted approach. This method is free from the usage of toxic and hazardous chemicals making it an environment friendly route. The synthesized bismuth sulphide is characterized using transmission electron microscope (TEM), powder X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and cyclic voltammetry (CV). From our experiments we find that bismuth sulphide produced using this bio-assisted approach exhibits a hexagonal shaped plate-like structures and is stabilized by the extracellular proteins present in the culture medium. - Graphical abstract: A green chemistry approach towards the synthesis of bismuth (III) sulphide nanostructures at room temperature using a hydrogen sulphide producing microorganism namely, Clostridium acetobutylicum is demonstrated. - Highlights: • Environmentally benign (greener) route towards synthesis of Bi 2 S 3 nanostructures. • Bio-assisted synthesis of Bi 2 S 3 at room temperature using Clostridium acetobutylicum. • Extracellular proteins in H 2 S producing microorganism as stabilizer for Bi 2 S 3 NPs. • Hexagonal platelets of Bi 2 S 3 possessing an orthorhombic crystalline structure

  3. Phospholipase C produced by Clostridium botulinum types C and D: comparison of gene, enzymatic, and biological activities with those of Clostridium perfringens alpha-toxin.

    Science.gov (United States)

    Fatmawati, Ni Nengah Dwi; Sakaguchi, Yoshihiko; Suzuki, Tomonori; Oda, Masataka; Shimizu, Kenta; Yamamoto, Yumiko; Sakurai, Jun; Matsushita, Osamu; Oguma, Keiji

    2013-01-01

    Clostridium botulinum type C and D strains recently have been found to produce PLC on egg yolk agar plates. To characterize the gene, enzymatic and biological activities of C. botulinum PLCs (Cb-PLCs), the cb-plc genes from 8 strains were sequenced, and 1 representative gene was cloned and expressed as a recombinant protein. The enzymatic and hemolytic activities of the recombinant Cb-PLC were measured and compared with those of the Clostridium perfringens alpha-toxin. Each of the eight cb-plc genes encoded a 399 amino acid residue protein preceded by a 27 residue signal peptide. The protein consists of 2 domains, the N- and C-domains, and the overall amino acid sequence identity between Cb-PLC and alpha-toxin was greater than 50%, suggesting that Cb-PLC is homologous to the alpha-toxin. The key residues in the N-domain were conserved, whereas those in the C-domain which are important in membrane interaction were different than in the alpha-toxin. As expected, Cb-PLC could hydrolyze egg yolk phospholipid, p-nitrophenylphosphorylcholine, and sphingomyelin, and also exhibited hemolytic activity;however, its activities were about 4- to over 200-fold lower than those of alpha-toxin. Although Cb-PLC showed weak enzymatic and biological activities, it is speculated that Cb-PLC might play a role in the pathogenicity of botulism or for bacterial survival.

  4. Antibiotic treatment for Clostridium difficile-associated diarrhoea in adults.

    Science.gov (United States)

    Nelson, Richard L; Suda, Katie J; Evans, Charlesnika T

    2017-03-03

    Clostridium difficile (C. difficile) is recognized as a frequent cause of antibiotic-associated diarrhoea and colitis. This review is an update of a previously published Cochrane review. The aim of this review is to investigate the efficacy and safety of antibiotic therapy for C. difficile-associated diarrhoea (CDAD), or C. difficile infection (CDI), being synonymous terms. We searched MEDLINE, EMBASE, CENTRAL and the Cochrane IBD Group Specialized Trials Register from inception to 26 January 2017. We also searched clinicaltrials.gov and clinicaltrialsregister.eu for ongoing trials. Only randomised controlled trials assessing antibiotic treatment for CDI were included in the review. Three authors independently assessed abstracts and full text articles for inclusion and extracted data. The risk of bias was independently rated by two authors. For dichotomous outcomes, we calculated the risk ratio (RR) and corresponding 95% confidence interval (95% CI). We pooled data using a fixed-effect model, except where significant heterogeneity was detected, at which time a random-effects model was used. The following outcomes were sought: sustained symptomatic cure (defined as initial symptomatic response and no recurrence of CDI), sustained bacteriologic cure, adverse reactions to the intervention, death and cost. Twenty-two studies (3215 participants) were included. The majority of studies enrolled patients with mild to moderate CDI who could tolerate oral antibiotics. Sixteen of the included studies excluded patients with severe CDI and few patients with severe CDI were included in the other six studies. Twelve different antibiotics were investigated: vancomycin, metronidazole, fusidic acid, nitazoxanide, teicoplanin, rifampin, rifaximin, bacitracin, cadazolid, LFF517, surotomycin and fidaxomicin. Most of the studies were active comparator studies comparing vancomycin with other antibiotics. One small study compared vancomycin to placebo. There were no other studies that

  5. Process engineering and scale-up of autotrophic Clostridium strain P11 syngas fermentation

    Science.gov (United States)

    Kundiyana, Dimple Kumar Aiyanna

    Scope and Method of Study. Biomass gasification followed by fermentation of syngas to ethanol is a potential process to produce bioenergy. The process is currently being researched under laboratory- and pilot-scale in an effort to optimize the process conditions and make the process feasible for commercial production of ethanol and other biofuels such as butanol and propanol. The broad research objectives for the research were to improve ethanol yields during syngas fermentation and to design a economical fermentation process. The research included four statistically designed experimental studies in serum bottles, bench-scale and pilot-scale fermentors to screen alternate fermentation media components, to determine the effect of process parameters such as pH, temperature and buffer on syngas fermentation, to determine the effect of key limiting nutrients of the acetyl-CoA pathway in a continuous series reactor design, and to scale-up the syngas fermentation in a 100-L pilot scale fermentor. Findings and Conclusions. The first experimental study identified cotton seed extract (CSE) as a feasible medium for Clostridium strain P11 fermentation. The study showed that CSE at 0.5 g L-1 can potentially replace all the standard Clostridium strain P11 fermentation media components while using a media buffer did not significantly improve the ethanol production when used in fermentation with CSE. Scale-up of the CSE fermentation in 2-L and 5-L stirred tank fermentors showed 25% increase in ethanol yield. The second experimental study showed that syngas fermentation at 32°C without buffer was associated with higher ethanol concentration and reduced lag time in switching to solventogenesis. Conducting fermentation at 40°C or by lowering incubation pH to 5.0 resulted in reduced cell growth and no production of ethanol or acetic acid. The third experiment studied the effect of three limiting nutrients, calcium pantothenate, vitamin B12 and CoCl2 on syngas fermentation. Results

  6. Microencapsulation of Clostridium difficile specific bacteriophages using microfluidic glass capillary devices for colon delivery using pH triggered release.

    Directory of Open Access Journals (Sweden)

    Gurinder K Vinner

    Full Text Available The prevalence of pathogenic bacteria acquiring multidrug antibiotic resistance is a global health threat to mankind. This has motivated a renewed interest in developing alternatives to conventional antibiotics including bacteriophages (viruses as therapeutic agents. The bacterium Clostridium difficile causes colon infection and is particularly difficult to treat with existing antibiotics; phage therapy may offer a viable alternative. The punitive environment within the gastrointestinal tract can inactivate orally delivered phages. C. difficile specific bacteriophage, myovirus CDKM9 was encapsulated in a pH responsive polymer (Eudragit® S100 with and without alginate using a flow focussing glass microcapillary device. Highly monodispersed core-shell microparticles containing phages trapped within the particle core were produced by in situ polymer curing using 4-aminobenzoic acid dissolved in the oil phase. The size of the generated microparticles could be precisely controlled in the range 80 μm to 160 μm through design of the microfluidic device geometry and by varying flow rates of the dispersed and continuous phase. In contrast to free 'naked' phages, those encapsulated within the microparticles could withstand a 3 h exposure to simulated gastric fluid at pH 2 and then underwent a subsequent pH triggered burst release at pH 7. The significance of our research is in demonstrating that C. difficile specific phage can be formulated and encapsulated in highly uniform pH responsive microparticles using a microfluidic system. The microparticles were shown to afford significant protection to the encapsulated phage upon prolonged exposure to an acid solution mimicking the human stomach environment. Phage encapsulation and subsequent release kinetics revealed that the microparticles prepared using Eudragit® S100 formulations possess pH responsive characteristics with phage release triggered in an intestinal pH range suitable for therapeutic

  7. Surface-layer protein A (SlpA is a major contributor to host-cell adherence of Clostridium difficile.

    Directory of Open Access Journals (Sweden)

    Michelle M Merrigan

    Full Text Available Clostridium difficile is a leading cause of antibiotic-associated diarrhea, and a significant etiologic agent of healthcare-associated infections. The mechanisms of attachment and host colonization of C. difficile are not well defined. We hypothesize that non-toxin bacterial factors, especially those facilitating the interaction of C. difficile with the host gut, contribute to the initiation of C. difficile infection. In this work, we optimized a completely anaerobic, quantitative, epithelial-cell adherence assay for vegetative C. difficile cells, determined adherence proficiency under multiple conditions, and investigated C. difficile surface protein variation via immunological and DNA sequencing approaches focused on Surface-Layer Protein A (SlpA. In total, thirty-six epidemic-associated and non-epidemic associated C. difficile clinical isolates were tested in this study, and displayed intra- and inter-clade differences in attachment that were unrelated to toxin production. SlpA was a major contributor to bacterial adherence, and individual subunits of the protein (varying in sequence between strains mediated host-cell attachment to different extents. Pre-treatment of host cells with crude or purified SlpA subunits, or incubation of vegetative bacteria with anti-SlpA antisera significantly reduced C. difficile attachment. SlpA-mediated adherence-interference correlated with the attachment efficiency of the strain from which the protein was derived, with maximal blockage observed when SlpA was derived from highly adherent strains. In addition, SlpA-containing preparations from a non-toxigenic strain effectively blocked adherence of a phylogenetically distant, epidemic-associated strain, and vice-versa. Taken together, these results suggest that SlpA plays a major role in C. difficile infection, and that it may represent an attractive target for interventions aimed at abrogating gut colonization by this pathogen.

  8. Heat shock and prolonged heat stress attenuate neurotoxin and sporulation gene expression in group I Clostridium botulinum strain ATCC 3502.

    Science.gov (United States)

    Selby, Katja; Mascher, Gerald; Somervuo, Panu; Lindström, Miia; Korkeala, Hannu

    2017-01-01

    Foodborne pathogenic bacteria are exposed to a number of environmental stresses during food processing, storage, and preparation, and in the human body. In order to improve the safety of food, the understanding of molecular stress response mechanisms foodborne pathogens employ is essential. Many response mechanisms that are activated during heat shock may cross-protect bacteria against other environmental stresses. To better understand the molecular mechanisms Clostridium botulinum, the causative agent of botulism, utilizes during acute heat stress and during adaptation to stressfully high temperature, the C. botulinum Group I strain ATCC 3502 was grown in continuous culture at 39°C and exposed to heat shock at 45°C, followed by prolonged heat stress at 45°C to allow adaptation of the culture to the high temperature. Growth in continuous culture was performed to exclude secondary growth phase effects or other environmental impacts on bacterial gene transcription. Changes in global gene expression profiles were studied using DNA microarray hybridization. During acute heat stress, Class I and III heat shock genes as well as members of the SOS regulon were activated. The neurotoxin gene botA and genes encoding the neurotoxin-associated proteins were suppressed throughout the study. Prolonged heat stress led to suppression of the sporulation machinery whereas genes related to chemotaxis and motility were activated. Induced expression of a large proportion of prophage genes was detected, suggesting an important role of acquired genes in the stress resistance of C. botulinum. Finally, changes in the expression of a large number of genes related to carbohydrate and amino acid metabolism indicated remodeling of the cellular metabolism.

  9. Identification of patients at high risk for Clostridium difficile infection : Development and validation of a risk prediction model in hospitalized patients treated with antibiotics

    NARCIS (Netherlands)

    van Werkhoven, C. H.; van der Tempel, J.; Jajou, R.; Thijsen, S. F T; Diepersloot, R. J A; Bonten, M. J M; Postma, D. F.; Oosterheert, J. J.

    2015-01-01

    To develop and validate a prediction model for Clostridium difficile infection (CDI) in hospitalized patients treated with systemic antibiotics, we performed a case-cohort study in a tertiary (derivation) and secondary care hospital (validation). Cases had a positive Clostridium test and were

  10. Priority of the genus name Clostridium Prazmowski 1880 (Approved Lists 1980) vs Sarcina Goodsir 1842 (Approved Lists 1980) and the creation of the illegitimate combinations Clostridium maximum (Lindner 1888) Lawson and Rainey 2016 and Clostridium ventriculi (Goodsir 1842) Lawson and Rainey 2016 that may not be used.

    Science.gov (United States)

    Tindall, B J

    2016-11-01

    In a recent publication that attempts to deal with the growing problem of taxa being added to the genus Clostridium that are outside of Clostridium (16S rRNA) group I, a solution is proposed that seeks to limit the genus Clostridium Prazmowski 1880 (Approved Lists 1980) to a small number of species 'related' to the type species, Clostridium butyricum Prazmowski 1880 (Approved Lists 1980). It has been proposed that this genus should also include members of the genus Sarcina Goodsir 1842 (Approved Lists 1980), Sarcinamaxima Lindner 1888 (Approved Lists 1980) and Sarcinaventriculi Goodsir 1842 (Approved Lists 1980), the latter being the nomenclatural type of the genus Sarcina Goodsir 1842 (Approved Lists 1980). In making proposals to treat the genus name Sarcina Goodsir 1842 (Approved Lists 1980) as a synonym of ClostridiumPrazmowski 1880 (Approved Lists 1980), reference is made to the wording of the International Code of Nomenclature of Bacteria. However, while that wording is factually correct, other parts of the Code are relevant to this issue and clearly indicate that the proposed course of action is not sanctioned by texts that have not been directly made reference to. Rather than avoiding confusion it has been contributed to, and it is necessary to document where the problems lie.

  11. Effect of Clostridium butyricum supplementation on the development of intestinal flora and the immune system of neonatal mice.

    Science.gov (United States)

    Miao, Rui-Xue; Zhu, Xin-Xin; Wan, Chao-Min; Wang, Zhi-Ling; Wen, Yang; Li, Yi-Yuan

    2018-01-01

    The objective of the present study was to examine whether Clostridium butyricum supplementation has a role in the regulation of the intestinal flora and the development of the immune system of neonatal mice. A total of 30 pregnant BALB/c mice, including their offspring, were randomly divided into three groups: In the maternal intervention group (Ba), maternal mice were treated with Clostridium butyricum from birth until weaning at postnatal day 21 (PD21) followed by administration of saline to the offspring at PD21-28; in the offspring intervention group (Ab), breast-feeding maternal mice were supplemented with saline and offspring were directly supplemented with Clostridium butyricum from PD21-28; in the both maternal and offspring intervention group (Bb), both maternal mice and offspring were supplemented with Clostridium butyricum at PD 0-21 and at PD21-28. While mice in the control group were given the same volume of normal saline. Stool samples from the offspring were collected at PD14, -21 and -28 to observe the intestinal flora by colony counts of Enterococcus spp., Enterobacter spp., Bifidobacterium spp. and Lactobacillus spp. Detection of intestinal secreted immunoglobulin A (sIgA) levels and serum cytokine (interferon-γ, and interleukin-12, -4 and -10) levels in offspring was performed to evaluate the effect on their immune system. The results revealed that compared with the control group, offspring in the Ba group displayed significantly decreased stool colony counts of Enterococcus spp. (t=3.123, Pflora balance in their offspring. However, due to insignificant effects on sIgA level and the associated cytokines, Clostridium butyricum had a limited influence on the balance of type 1 vs. type 2 T-helper cells. However, using Clostridium butyricum as an invention may be a safe method for improving the balance of intestinal flora and associated processes in offspring.

  12. Sensitive assays enable detection of serum IgG antibodies against Clostridium difficile toxin A and toxin B in healthy subjects and patients with Clostridium difficile infection.

    Science.gov (United States)

    Zhao, Xuemei; Bender, Florent; Shukla, Rajiv; Kang, John J; Caro-Aguilar, Ivette; Laterza, Omar F

    2016-04-01

    Pathogenic Clostridium difficile produces two proinflammatory exotoxins, toxin A and toxin B. Low level of serum antitoxin IgG antibodies is a risk factor for the development of primary and recurrent C. difficile infection (CDI). We developed and validated two sensitive, titer-based electrochemiluminescence assays for the detection of serum antibody levels against C. difficile toxins A and B. These assays demonstrated excellent precision. The sensitivity of the assays allowed the detection of antitoxin A and antitoxin B IgG antibodies in all tested serum samples during assay validation. The validated titer-based assays enable assessment of antitoxin A and antitoxin B IgG antibodies as potential biomarkers to identify patients with CDI at increased risk for CDI recurrence.

  13. Impact of a prevention bundle on Clostridium difficile infection rates in a hospital in the Southeastern United States.

    Science.gov (United States)

    Davis, Bionca M; Yin, Jingjing; Blomberg, Doug; Fung, Isaac Chun-Hai

    2016-12-01

    We sought to assess the impact of a multicomponent prevention program on hospital-acquired Clostridium difficile infections in a hospital in the Southeastern United States. We collected retrospective data of 140 patients from years 2009-2014 and applied the Poisson regression model for analysis. We did not find any significant associations of increased risk of Clostridium difficile infections for the preintervention group. Further studies are needed to test multifaceted bundles in hospitals with high infection rates. Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

  14. Hand hygiene for the management of a patient infected with Clostridium difficile in the presence of hospital infections

    Directory of Open Access Journals (Sweden)

    Krystyna Nowacka

    2017-06-01

    Such pathogens include Clostridium difficile. That is the most important pathogen causing diarrhea.  Stem infection of Clostridium difficile may cause serious diseases and medical conditions, particularly in the elderly, debilitated as a result of chronic diseases. The need to respect the principles of hand hygiene by medical staff is widely recognized. Causes of irregularities in the field of hand hygiene are different, for example. the intensity of the work, insufficient medical knowledge, limited access to devices for effective hand hygiene or ignorance of hand disinfection techniques.

  15. Identification and Characterization of a New Enterotoxin Produced by Clostridium perfringens Isolated from Food Poisoning Outbreaks.

    Directory of Open Access Journals (Sweden)

    Daisuke Irikura

    Full Text Available There is a strain of Clostridium perfringens, W5052, which does not produce a known enterotoxin. We herein report that the strain W5052 expressed a homologue of the iota-like toxin components sa and sb of C. spiroforme, named Clostridium perfringens iota-like enterotoxin, CPILE-a and CPILE-b, respectively, based on the results of a genome sequencing analysis and a systematic protein screening. In the nicotinamide glyco-hydrolase (NADase assay the hydrolysis activity was dose-dependently increased by the concentration of rCPILE-a, as judged by the mass spectrometry analysis. In addition, the actin monomer of the lysates of Vero and L929 cells were radiolabeled in the presence of [32P]NAD and rCPILE-a. These findings indicated that CPILE-a possesses ADP-ribosylation activity. The culture supernatant of W5052 facilitated the rounding and killing of Vero and L929 cells, but the rCPILE-a or a non-proteolyzed rCPILE-b did not. However, a trypsin-treated rCPILE-b did. Moreover, a mixture of rCPILE-a and the trypsin-treated rCPILE-b enhanced the cell rounding and killing activities, compared with that induced by the trypsin-treated rCPILE-b alone. The injection of the mixture of rCPILE-a and the trypsin-treated rCPILE-b into an ileum loop of rabbits evoked the swelling of the loop and accumulation of the fluid dose-dependently, suggesting that CPILE possesses enterotoxic activity. The evidence presented in this communication will facilitate the epidemiological, etiological, and toxicological studies of C. perfringens food poisoning, and also stimulate studies on the transfer of the toxins' gene(s among the Genus Clostridium.

  16. Monitoring Clostridium difficile infection in an acute hospital: prevalence or incidence studies?

    LENUS (Irish Health Repository)

    Lavan, A H

    2012-02-15

    BACKGROUND: Surveillance of Clostridium difficile infection (CDI) is an essential component of a CDI preventative programme. AIMS: The aim of this study was to evaluate two methods of CDI surveillance. METHODS: Prevalence of CDI, antibiotic use and associated co-morbidity was assessed weekly on two wards over 6 weeks. In addition, CDI incidence surveillance was performed on all new CDI cases over a 13-week period. Cases were assessed for CDI risk factors, disease severity, response to treatment and outcome at 6 months. RESULTS: Clostridium difficile infection prevalence was 3.5% (range 2.9-6.1%) on the medical ward and 1.1% (range 0-3.5%) on the surgical ward. Patients on the medical ward were older and more likely to be colonised with MRSA; however, recent antibiotic use was more prevalent among surgical patients. Sixty-one new CDI cases were audited. Patients were elderly (mean age 71 years) with significant co-morbidity (median age adjusted Charlson co-morbidity score 5). CDI ribotypes included 027 (29 cases) 078 (5) and 106 (4). Eight patients developed severe CDI, seven due to 027. Antibiotic use was common with 56% receiving three or more antibiotics in the preceding 8 weeks. Twenty-four patients had died at 6 months, five due to CDI. CONCLUSION: Clostridium difficile infection prevalence gives a broad overview of CDI and points to areas that require more detailed surveillance and requires little time. However, patient-based CDI incidence surveillance provides a more useful analysis of CDI risk factors, disease and outcome for planning preventative programmes and focusing antibiotic stewardship efforts.

  17. Parameters for the mathematical modelling of Clostridium difficile acquisition and transmission: a systematic review.

    Directory of Open Access Journals (Sweden)

    Eroboghene H Otete

    Full Text Available INTRODUCTION: Mathematical modelling of Clostridium difficile infection dynamics could contribute to the optimisation of strategies for its prevention and control. The objective of this systematic review was to summarise the available literature specifically identifying the quantitative parameters required for a compartmental mathematical model of Clostridium difficile transmission. METHODS: Six electronic healthcare databases were searched and all screening, data extraction and study quality assessments were undertaken in duplicate. Results were synthesised using a narrative approach. RESULTS: Fifty-four studies met the inclusion criteria. Reproduction numbers for hospital based epidemics were described in two studies with a range from 0.55 to 7. Two studies provided consistent data on incubation periods. For 62% of cases, symptoms occurred in less than 4 weeks (3-28 days after infection. Evidence on contact patterns was identified in four studies but with limited data reported for populating a mathematical model. Two studies, including one without clinically apparent donor-recipient pairs, provided information on serial intervals for household or ward contacts, showing transmission intervals of <1 week in ward based contacts compared to up to 2 months for household contacts. Eight studies reported recovery rates of between 75%-100% for patients who had been treated with either metronidazole or vancomycin. Forty-nine studies gave recurrence rates of between 3% and 49% but were limited by varying definitions of recurrence. No study was found which specifically reported force of infection or net reproduction numbers. CONCLUSIONS: There is currently scant literature overtly citing estimates of the parameters required to inform the quantitative modelling of Clostridium difficile transmission. Further high quality studies to investigate transmission parameters are required, including through review of published epidemiological studies where these

  18. Alpha toxin specific PCR for detection of toxigenic strains of Clostridium perfringens in Poultry

    Directory of Open Access Journals (Sweden)

    Malmarugan Shanmugasamy

    2012-12-01

    Full Text Available Aim : Isolation of clostridium perfirngens from necrotic enteritis cases in poultry and confirmation by alpha toxin specific PCR Materials and methods: Robertson cooked meat medium with Brain Heart Infusion broth was used for isolation of C. perfringens from intestinal contents of necrotic enteritis suspected birds. Positive cultures from perfringens agar were further confirmed by biochemical tests and subjected to alpha toxin specific PCR. Results: Twenty Clostridium perfringens isolates were isolated from intestinal contents of thirty five NE suspected birds. Out of the twenty isolates, fourteen were isolated from commercial broilers of 2 to 6 wk of age and six from commercial layers of 9 to 15 wk of age. Frequency of isolation of C. perfringens was more with Robertson cooked meat medium with BHI broth than thioglycollate broth alone. When positive cultures were streaked on to clostridial agar appreciable luxuriant growths were obtained and the selective streaking of these colonies on perfringens agar with supplements revealed rough and black colonies with sulphate reduction. The isolates produced rough and black colonies with sulphate reduction on perfringens agar, double zone haemolysis on sheep blood agar, stormy clot fermentation on milk medium and opalescence on egg yolk medium. The isolates were found negative for oxidase, catalase, liquefied gelatin, fermented glucose, maltose, lactose and sucrose except mannitol. All the fourteen isolates obtained from commercial broilers proved the alpha toxin producing strains of C. perfringens when they were subjected to alpha toxin specific PCR. Conclusion : This study revealed alpha toxin specific PCR is highly useful for detection of toxigenic strains of Clostridium perfringens in poultry [Vet. World 2012; 5(6.000: 365-368

  19. Preparing for Surgery

    Science.gov (United States)

    ... Events Advocacy For Patients About ACOG Preparing for Surgery Home For Patients Search FAQs Preparing for Surgery ... Surgery FAQ080, August 2011 PDF Format Preparing for Surgery Gynecologic Problems What is the difference between outpatient ...

  20. Induction and regeneration of autoplasts from Clostridium thermohydrosulfuricum JW102 and Thermoanaerobacter ethanolicus JW200

    Energy Technology Data Exchange (ETDEWEB)

    Peteranderl, R.; Canganella, F.; Holzenburg, A.; Wiegel, J. [Univ. of Georgia, Athens, GA (United States)

    1993-10-01

    Autolysis was induced to form stable, cell wall-free cells of Clostridium thermohydrosulfuricum JW102 and Thermoanaerobacter ethanolicus JW200, using a complex medium containing glycine (0.4% wt/vol) and/or sucrose or glycerol (10% wt/vol) at an optimum temperature of 64{degrees}C. Autoplasts of both bacteria were grown as L-phase colonies on solid medium; more than 50% of these colonies regenerated to the walled form during prolonged incubation. The removal of the cell wall was confirmed by electron microscopy.

  1. The Potential Value of Clostridium difficile Vaccine: An Economic Computer Simulation Model

    Science.gov (United States)

    Lee, Bruce Y.; Popovich, Michael J.; Tian, Ye; Bailey, Rachel R.; Ufberg, Paul J.; Wiringa, Ann E.; Muder, Robert R.

    2010-01-01

    Efforts are currently underway to develop a vaccine against Clostridium difficile infection (CDI). We developed two decision analytic Monte Carlo computer simulation models: (1) an Initial Prevention Model depicting the decision whether to administer C. difficile vaccine to patients at-risk for CDI and (2) a Recurrence Prevention Model depicting the decision whether to administer C. difficile vaccine to prevent CDI recurrence. Our results suggest that a C. difficile vaccine could be cost-effective over a wide range of C. difficile risk, vaccine costs, and vaccine efficacies especially when being used post-CDI treatment to prevent recurrent disease. PMID:20541582

  2. Faecal excretion of brush border membrane enzymes in patients with clostridium difficile diarrhoea

    Directory of Open Access Journals (Sweden)

    Katyal R

    2002-01-01

    Full Text Available PURPOSE: To look for the presence of intestinal brush border membrane (BBM enzymes in the faecal samples of patients with Clostridium difficile association. METHODS: One hundred faecal samples were investigated for C.difficile toxin (CDT. Simultaneous assays for faecal excretion of intestinal BBM enzymes viz., disaccharidases, alkaline phosphatase (AP and leucine aminopeptidase (LAP were also done. RESULTS: C.difficile toxin was detected in 25 (25% of the samples with a titre ranging from 10 to 160. No significant difference (p>0.05 was seen between the CDT positive and negative groups with any of the disaccharidases studied. However, significant increase (pC.difficile diarrhoea.

  3. Detection of Clostridium difficile in Retail Ground Meat Products in Manitoba

    Directory of Open Access Journals (Sweden)

    Monique Visser

    2012-01-01

    Full Text Available The aim of the present study was to determine whether Clostridium difficile was present in uncooked retail ground beef and ground pork products sold in Winnipeg, Manitoba. Using an alcohol treatment protocol and inoculation of cultures on C difficile Moxalactam Norfloxacin (CDMN, toxigenic C difficile was found in 6.3% of 48 meat samples. The C difficile isolates belonged to different pulsotypes, all of which had been previously isolated from the stool of Manitoba patients with C difficile disease. Because cooking of meat will not eradicate C difficile spores, this raises a concern regarding potential foodborne transmissibility of this organism.

  4. Molecular analysis of the interaction between Clostridium perfringens Enterotoxin and Claudins

    OpenAIRE

    Protze, Jonas

    2015-01-01

    Claudins are essential constituents of Tight Junctions (TJs) and responsible for maintenance of these cell-cell contacts. Binding of Clostridium perfringens Enterotoxin’s C-terminal domain (cCPE) to the extracellular loop 2 (EZS2) of claudins, especially Cld3, Cld4 and Cld6-Cld9 causes a reversible opening of TJs. Thus, a structure-function analysis of this system is relevant for biomedical application, since cCPE could be used to enhance paracellular drug uptake. Furthermore cCPE respectivel...

  5. Prospects for the Elimination of Clostridium Botulinum from Fish and Fishery Products by Irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Hobbs, G. [Torry Research Station, Ministry of Technology, Aberdeen, Scotland (United Kingdom)

    1968-07-15

    The paper first discusses the occurrence of Clostridium botulinum in fish and fishery products and their resistance to irradiation, and then considers the different possible irradiation treatments such as radappertization, radurization and combined processes. For Great Britain it is suggested that sea- frozen fish should be thawed, filleted, packaged and irradiated at 0.3 Mrads. This irradiation treatment would give a sufficient extension of the market life. To eliminate the botulism hazard a temperature of less than 5 Degree-Sign C for storage and distribution is recommended. (author)

  6. Impact of sink location on hand hygiene compliance for Clostridium difficile infection.

    Science.gov (United States)

    Zellmer, Caroline; Blakney, Rebekah; Van Hoof, Sarah; Safdar, Nasia

    2015-04-01

    Hand hygiene with soap and water after the care of a patient with Clostridium difficile infection is essential to reduce nosocomial transmission in an outbreak situation. Factors that may pose barriers to user completion of infection prevention measures, such as hand hygiene, are of interest. We undertook a quantitative study to evaluate the relationship between sink location and compliance with handwashing among health care workers and visitors in a surgical transplant unit. We found that placement of 2 more easily visible sinks in a surgical transplant unit was associated with improved adherence to handwashing. Published by Elsevier Inc.

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

    Science.gov (United States)

    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

  8. Clostridium difficile infection: Early history, diagnosis and molecular strain typing methods.

    Science.gov (United States)

    Rodriguez, C; Van Broeck, J; Taminiau, B; Delmée, M; Daube, G

    2016-08-01

    Recognised as the leading cause of nosocomial antibiotic-associated diarrhoea, the incidence of Clostridium difficile infection (CDI) remains high despite efforts to improve prevention and reduce the spread of the bacterium in healthcare settings. In the last decade, many studies have focused on the epidemiology and rapid diagnosis of CDI. In addition, different typing methods have been developed for epidemiological studies. This review explores the history of C. difficile and the current scope of the infection. The variety of available laboratory tests for CDI diagnosis and strain typing methods are also examined. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. A case of multiple recurrence of Clostridium difficile infection with severe hematochezia in an immunocompromised host.

    Science.gov (United States)

    Zhang, Xuewu; Chen, Yunbo; Gu, Silan; Zheng, Beiwen; Lv, Tao; Lou, Yinjun; Jin, Jie

    2016-12-01

    Clostridium difficile infection (CDI) is increasing in incidence and severity. Clinically, diarrhea frequently occurs, but severe hematochezia is rarely seen with CDI. We describe here a hematopoietic stem cell transplantation (HSCT) recipient who experienced life-threatening gastrointestinal bleeding due to severe CDI. Subsequent stool surveillance and molecular typing observed the patient who had two episodes of recurrence with a new strain of C. difficile distinct from the initial infection. We analyze C. difficile strains obtained from the patient, and also discuss the diagnosis and treatment of this case. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Fæcestransplantation som behandling af Clostridium difficile-infektion, colitis ulcerosa og metabolisk syndrom

    DEFF Research Database (Denmark)

    Carstensen, Jeppe West; Hansen, Axel Kornerup

    2014-01-01

    Faecal transplantation as a treatment for Clostridium difficile infection, ulcerative colitis and the metabolic syndrome Faecal transplantation as a therapeutic tool is increasingly reported in the scientific literature. Faecal transplantation is currently becoming a treatment for nosocomial......, refractory infections with C. difficile. Furthermore, faecal transplantation has been suggested as a treatment for ulcerative colitis as well as for the metabolic syndrome. In the accumulated literature faecal transplantations appear to be safe, effective and superior to current treatments. Faecal...... transplantation remains a sparsely investigated treatment, however, especially for other diagnoses than C. difficile infection....

  11. Evaluation of DNA Extraction Methods Suitable for PCR-based Detection and Genotyping of Clostridium botulinum

    DEFF Research Database (Denmark)

    Auricchio, Bruna; Anniballi, Fabrizio; Fiore, Alfonsina

    2013-01-01

    in terms of cost, time, labor, and supplies. Eleven botulinum toxin–producing clostridia strains and 25 samples (10 food, 13 clinical, and 2 environmental samples) naturally contaminated with botulinum toxin–producing clostridia were used to compare 4 DNA extraction procedures: Chelex® 100 matrix, Phenol......Sufficient quality and quantity of extracted DNA is critical to detecting and performing genotyping of Clostridium botulinum by means of PCR-based methods. An ideal extraction method has to optimize DNA yield, minimize DNA degradation, allow multiple samples to be extracted, and be efficient...

  12. Antibiotic-associated diarrhoea, Clostridium difficile, and short-chain fatty acids

    DEFF Research Database (Denmark)

    Hove, H; Tvede, M; Mortensen, P B

    1996-01-01

    BACKGROUND: It has been hypothesized that Clostridium difficile and decreased colonic production of short-chain fatty acids (SCFAs) cause the development of antibiotic-associated diarrhoea. We therefore wanted to investigate the effects of an intensive and uniform antibiotic therapy on faecal SCFAs...... concentrations. C. difficile, and extent of diarrhoea. METHODS: Fifteen liver-transplanted patients who received oral bowel flora suppression therapy (6.3 g cefuroxime, 0.6 g tobramycin, and 0.5 g nystatin three times daily) were studied for 12 days before and 12 days after discontinuation of therapy. RESULTS...

  13. Alphitobius diaperinus spp como veiculador de Clostridium perfringens em granjas avícolas do interior paulista - Brasil Alphitobius diaperinus spp as a vector of Clostridium perfringens in broiler houses in the state of São Paulo - Brazil

    Directory of Open Access Journals (Sweden)

    Juliano Vittori

    2007-06-01

    Full Text Available O besouro Alphitobius diaperinus spp (cascudinho é visto como uma importante praga da avicultura mundial. Por suas características comportamentais e hábitos biológicos que dificultam seu controle, é considerado um vetor de agentes patogênicos. O objetivo desta pesquisa foi investigar o cascudinho como possível vetor de Clostridium perfringens em granjas avícolas industriais, localizadas em diferentes regiões do interior Paulista. Através de métodos bacteriológicos convencionais, em 40 amostras analisadas, foram encontradas contagens significativas de Clostridium perfringens em todas elas. A partir dos resultados obtidos, pôde-se demonstrar o potencial deste inseto como vetor do agente responsável pela enterite necrótica.The Alphitobius diaperinus spp (lesser mealworm is considered an important world poultry plague. Due to its behavior characteristics and biological habits that make its control difficult it is considered a vector of pathogenic agents. The objective of this research was to investigate the little mealworm as possible vector of Clostridium perfringens in broiler houses, located in different parts of the state of São Paulo. Through conventional bacteriological methods, 40 samples of little mealworm collected were analyzed. Clostridium perfringens was found in all of the samples and the potential of this insect as vector of the necrotic enteritis was demonstrated.

  14. A novel regulator controls Clostridium difficile sporulation, motility and toxin production.

    Science.gov (United States)

    Edwards, Adrianne N; Tamayo, Rita; McBride, Shonna M

    2016-06-01

    Clostridium difficile is an anaerobic pathogen that forms spores which promote survival in the environment and transmission to new hosts. The regulatory pathways by which C. difficile initiates spore formation are poorly understood. We identified two factors with limited similarity to the Rap sporulation proteins of other spore-forming bacteria. In this study, we show that disruption of the gene CD3668 reduces sporulation and increases toxin production and motility. This mutant was more virulent and exhibited increased toxin gene expression in the hamster model of infection. Based on these phenotypes, we have renamed this locus rstA, for regulator of sporulation and toxins. Our data demonstrate that RstA is a bifunctional protein that upregulates sporulation through an unidentified pathway and represses motility and toxin production by influencing sigD transcription. Conserved RstA orthologs are present in other pathogenic and industrial Clostridium species and may represent a key regulatory protein controlling clostridial sporulation. © 2016 John Wiley & Sons Ltd.

  15. Biohydrogen production from a novel alkalophilic isolate Clostridium sp. IODB-O3.

    Science.gov (United States)

    Patel, Anil Kumar; Debroy, Arundhati; Sharma, Sandeep; Saini, Reetu; Mathur, Anshu; Gupta, Ravi; Tuli, Deepak Kumar

    2015-01-01

    Hydrogen producing bacteria IODB-O3 was isolated from sludge and identified as Clostridium sp. by 16S rDNA gene analysis. In this study, biohydrogen production process was developed using low-cost agro-waste. Maximum H2 was produced at 37°C and pH 8.5. Maximum H2 yield was obtained 2.54±0.2mol-H2/mol-reducing sugar from wheat straw pre-hydrolysate (WSPH) and 2.61±0.1mol-H2/mol-reducing sugar from pre-treated wheat straw enzymatic-hydrolysate (WSEH). The cumulative H2 production (ml/L), 3680±105 and 3270±100, H2 production rate (ml/L/h), 153±5 and 136±5, and specific H2 production (ml/g/h), 511±5 and 681±10 with WSPH and WSEH were obtained, respectively. Biomass pre-treatment via steam-explosion generates ample amount of WSPH which remains unutilized for bioethanol production due to non-availability of efficient C5-fermenting microorganisms. This study shows that Clostridium sp. IODB-O3 is capable of utilizing WSPH efficiently for biohydrogen production. This would lead to reduced economic constrain on the overall cellulosic ethanol process and also establish a sustainable biohydrogen production process. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Characterization of Clostridium perfringens isolated from mammals and birds from Guwahati city, India

    Directory of Open Access Journals (Sweden)

    Mafruza S Rahman

    2012-01-01

    Full Text Available Of the 102 samples collected from mammals and birds, both domestic and captive wild, 48 were found to be positive for Clostridium perfringens. Most of the mammal isolates (84.38% appeared to have been collected from clinically affected animals, while 33.33% of the bird samples were from clinically affected and 21.43% from apparently healthy birds infected with C. perfringens. Isolates revealed high sensitivity to ciprofloxacin, enrofloxacin and norfloxacin. Among the isolated C. perfringens, 30 (62.50% showed DNase production. Hemolytic activity was recorded in 14 (24.16% of the isolates and 28 (58.33% showed phospholipase C production. All the phospholipase C positive isolates revealed the presence of cpa gene encoding alpha (α toxin. Of the 102 samples collected from mammals and birds, both domestic and captive wild, 48 were found to be positive for Clostridium perfringens. Most of the mammal isolates (84.38% appeared to have been collected from clinically affected animals, while 33.33% of the bird samples were from clinically affected and 21.43% from apparently healthy birds infected with C. perfringens. Isolates revealed high sensitivity to ciprofloxacin, enrofloxacin and norfloxacin. Among the isolated C. perfringens, 30 (62.50% showed DNase production. Hemolytic activity was recorded in 14 (24.16% of the isolates and 28 (58.33% showed phospholipase C production. All the phospholipase C positive isolates revealed the presence of cpa gene encoding α toxin.

  17. Human alpha-defensin-1 protects cells from intoxication with Clostridium perfringens iota toxin.

    Science.gov (United States)

    Fischer, Stephan; Popoff, Michel R; Barth, Holger

    2018-03-01

    Iota toxin is produced by Clostridium perfringens type E strains and associated with diarrhea in cattle and lambs. This binary protein toxin comprises the enzyme component iota a (Ia), which ADP-ribosylates G-actin, and the separate transport component iota b (Ib), which delivers Ia into the cytosol of target cells. Ib binds to cell receptors and forms biologically active toxin complexes with Ia, which cause rounding of adherent cells due to the destruction of the actin cytoskeleton. Here, we report that the human peptide α-defensin-1 protects cultured cells including human colon cells from intoxication with iota toxin. In contrast, the related ß-defensin-1 had no effect, indicating a specific mode of action. The α-defensin-1 did not inhibit ADP-ribosylation of actin by Ia in vitro. Pretreatment of Ib with α-defensin-1 prior to addition of Ia prevented intoxication. Additionally, α-defensin-1 protected cells from cytotoxic effects mediated by Ib in the absence of Ia, implicating that α-defensin-1 interacts with Ib to prevent the formation of biologically active iota toxin on cells. In conclusion, the findings contribute to a better understanding of the functions of α-defensin-1 and suggest that this human peptide might be an attractive starting point to develop novel pharmacological options to treat/prevent diseases associated with iota toxin-producing Clostridium perfringens strains.

  18. A survey of traditional Iranian food products for contamination with toxigenic Clostridium botulinum

    Directory of Open Access Journals (Sweden)

    H.R. Tavakoli

    Full Text Available Summary: This study aimed to determine the rate of Clostridium botulinum contamination in some traditional Iranian food products (cheese, kashk and salted fish and evaluate the efficacy of the mouse bioassay method in detection of C. botulinum toxins in these foods. A total of 131 samples (57 cheese, 11 kashk and 63 salted fish were collected and examined to determine the rate of contamination by C. botulinum. Standard monovalent anti-toxins were used to determine the types of toxin. C. botulinum bacteria were detected in 4.58% of the examined samples (1.52% of cheese and 3.06% of salted fish samples. While no contamination was detected in the kashk samples, C. botulinum types A and E were found to be dominant in cheese and salted fish samples, respectively. These results indicate—some traditional Iranian foods may be contaminated with different types of C. botulinum, and the consumption of these products, either raw or cooked, may contribute to food-borne intoxications. Keywords: Clostridium botulinum, Botulinum toxin, Traditional foods

  19. Combination treatment of clostridium perfringens spores to freezing and/or gamma irradiation

    International Nuclear Information System (INIS)

    El-Fouly, M.Z.; El-Zawahry, Y.A.; Aziz, N.H.

    1985-01-01

    Freezing process alone caused relatively low decrease in viable count of suspended spores in minced meat while it decreased the spore numbers suspended in saline solution by more than one log cycle especially in case of the Egyptian strain. An abrupt decrease in viable counts of clostridium spores was observed by application dose of 1KGY either before or after freezing followed by gradual decrease of viable counts up to 15 KGY. The synergestic effect of combined treatment was clearly obvious for spores suspended in minced meat, which usually contains protective agents which increase the resistance of microorganisms against the separate treatment of radiation of freezing especially with spores of NCTC 8798 strain. Freezing the saline suspending medium before or after irradiation after the sensitivity of clostridium spores by only small extent and gave negative synergestic effect in some treatment. The percentages of injured spores due to the combined treatment were ranged between 15-100% of the viable counts. The percentage of injured spores tended to increase as the radiation dose levels increased

  20. Effects of anti-inflammatory drugs on fever and neutrophilia induced by Clostridium difficile toxin B

    Directory of Open Access Journals (Sweden)

    R. A. Cardoso

    1996-01-01

    Full Text Available This study investigated the ability of Clostridium difficile toxin B, isolated from the VPI 10463 strain, to induce fever and neutrophilia in rats. Intravenous injection of toxin B (0.005–0.5 μg/kg evoked a dose-dependent increase in body temperature. The febrile response to 0.5 μg/kg of the toxin started in 2.5 h, peaked at 5 h, and subsided fully within 24 h. Toxin B also induced a dosedependent neutrophilia. Pretreatment with indomethacin (2 mg/kg, i.p. did not affect the neutrophilia induced by toxin B, but significantly reduced the febrile response measured 4 to 8 h after toxin B injection. Dexamethasone (0.5 mg/ kg also markedly diminished the febrile response induced by toxin B. These results show that Clostridium difficile toxin B induced a febrile response susceptible to inhibition by dexamethasone and indomethacin. Furthermore, they suggest that prostaglandins are not involved in the neutrophilia caused by this toxin.

  1. Intra-abdominal hypertension in fulminant Clostridium difficile infection--an under-recognized treatable complication.

    Science.gov (United States)

    Oud, Lavi

    2010-09-01

    Clostridium difficile is the most common cause of nosocomial infectious diarrhea in adults, with recent reports of increased severity and case fatality. Intra-abdominal hypertension (IAH) and abdominal compartment syndrome (ACS) are increasingly recognized and treatable complications of severe illness in medical patients, and are independent predictors of mortality. Patients with severe Clostridium difficile infection (CDI) are at increased risk for IAH and ACS. However, ACS has been only rarely described in this population. We report a case of a 61 year-old morbidly obese, chronically ill, ventilator dependent patient, who developed fulminant CDI, including progressive colonic distension, acute renal failure and intra-abdominal fluid sequestration. Her clinical course worsened abruptly, with new shock, worsening hypoxic respiratory failure, increased peak airway pressures and reduced tidal volumes. Intra-abdominal pressure was 30 mm Hg. The patient was not considered a surgical candidate, was refractory to escalating non-surgical support, and died following withdrawal of life support. Although patients with fulminant CDI share many risk factors for IAH and ACS, these conditions were rarely reported in this population and are likely under recognized, as was the case with the present patient. Increased vigilance for IAH is needed in this at-risk population.

  2. Clostridium difficile in retail baskets, trolleys, conveyor belts, and plastic bags in Saudi Arabia.

    Science.gov (United States)

    Alqumber, Mohammed A

    2014-10-01

    To determine Clostridium difficile (C. difficile) prevalence on retail surfaces and shoppers plastic bags. From 20 June to 10 August 2011, in a cross-sectional epidemiological study, 17 supermarkets from 2 cities, Albaha and Altaif, Saudi Arabia were sampled. A total of 800 samples, which comprised 200 samples per surveyed surface, were studied. These included baskets, trolleys, conveyer belts, and outgoing shoppers' plastic bags. Clostridium difficile strains were isolated. The isolates were characterized using ribotyping and  polymerase chain reaction for the detection of toxin A (tcdA), toxin B (tcdB), binary toxin (cdtB), and toxin C (tcdC) genes. Susceptibility to antibiotics was determined on a Muller-Hinton agar with 5% sheep blood agar using E-tests. Overall, the C. difficile prevalence on sampled surfaces was 0.75%. The highest prevalence was found on retail baskets and trolleys, followed by plastic bags. A total of 5 different ribotypes were identified. Alterations in tcdC were detected in ribotype 027 and BT1. All the identified isolates were susceptible to vancomycin, but resistant to levofloxacin. In this study, C. difficile was present at a rate of 0.75% on supermarket surfaces. Spore disinfection of implicated surfaces may be necessary to control any community-acquired infections caused by this pathogen. 

  3. Bacillus subtilis and yeast cell wall improve the intestinal health of broilers challenged by Clostridium perfringens.

    Science.gov (United States)

    Li, Z; Wang, W; Lv, Z; Liu, D; Guo, Y

    2017-12-01

    1. The objective was to investigate the effects of Bacillus subtilis, yeast cell wall (YCW) and their combination on intestinal health of broilers challenged by Clostridium perfringens over a 21-d period. 2. Using a 5 × 2 factorial arrangement of treatments, 800 1-d-old male Cobb 500 broilers were used to study the effects of feed additives (without additive or with zinc bacitracin, B. subtilis, YCW, and the combination of B. subtilis and YCW), pathogen challenge (without or with Clostridium perfringens challenge), and their interactive effects. 3. C. perfringens infection increased intestinal lesions scores, damaged intestinal histomorphology, increased serum endotoxin concentration, cytokine mRNA expression and intestinal population of C. perfringens and Escherichia coli and decreased ileal bifidobacteria numbers. The 4 additives decreased serum endotoxin. Zinc bacitracin tended to decrease cytokine mRNA expression and the intestinal number of C. perfringens and E. coli. B. subtilis, YCW and their combination increased cytokine mRNA expression. B. subtilis and YCW decreased the number of C. perfringens and E. coli in the ileum, and their combination decreased pathogens numbers in the ileum and caecum. 4. In conclusion, B. subtilis, YCW and their combination improved the intestinal health of NE-infected broilers, and could be potential alternatives to antibiotics.

  4. Routine Treatment-Resistant Clostridium difficile Infection during Recovery from Myxedema

    Directory of Open Access Journals (Sweden)

    Jan K. Adamski

    2017-11-01

    Full Text Available Development of the extreme form of hypothyroidism defined as myxedema is very rare. Acute symptoms and their management have been described in detail previously. However, not much attention has been devoted to therapeutic challenges that are faced in the recovery phase of the treatment, especially pertaining to the gastrointestinal system. The link between myxedema and the appearance of severe Clostridium difficile infection (CDI has not been established so far. A 61-year-old woman with no significant medical record was admitted to hospital because of infected heel pressure and thyroid dysfunction. A week later, due to hypothermia, hypotension, and unconsciousness, she was transferred to the intensive care unit. The clinical picture and the results of laboratory tests confirmed diagnosis of myxedema. After the introduction of resuscitative measures and hormonal substitution, patient’s condition stabilized within 10 days. Due to concomitant sepsis, initially piperacillin/tazobactam and later cefuroxime were administered. After 20 days of antibiotic therapy, the patient developed CDI that was resistant to the routine mode of treatment. The clinical recovery was achieved only after a fecal microbiota transplantation procedure. The function of the digestive tract in myxedema is disturbed by gastric achlorydia and reduced peristalsis, which in turn can predispose the small intestine to overgrowth of bacteria. The use of antibiotics can additionally decrease the intestinal bacterial diversity, favoring the overgrowth of Clostridium difficile. The authors conclude that myxedema may increase the likelihood of a treatment-resistant form of CDI that requires the implementation of fecal microbiota transplantation.

  5. Improving isopropanol tolerance and production of Clostridium beijerinckii DSM 6423 by random mutagenesis and genome shuffling.

    Science.gov (United States)

    Gérando, H Máté de; Fayolle-Guichard, F; Rudant, L; Millah, S K; Monot, F; Ferreira, Nicolas Lopes; López-Contreras, A M

    2016-06-01

    Random mutagenesis and genome shuffling was applied to improve solvent tolerance and isopropanol/butanol/ethanol (IBE) production in the strictly anaerobic bacteria Clostridium beijerinckii DSM 6423. Following chemical mutagenesis with N-methyl-N-nitro-N-nitrosoguanidine (NTG), screening of putatively improved strains was done by submitting the mutants to toxic levels of inhibitory chemicals or by screening for their tolerance to isopropanol (>35 g/L). Suicide substrates, such as ethyl or methyl bromobutyrate or alcohol dehydrogenase inhibitors like allyl alcohol, were tested and, finally, 36 mutants were isolated. The fermentation profiles of these NTG mutant strains were characterized, and the best performing mutants were used for consecutive rounds of genome shuffling. Screening of strains with further enhancement in isopropanol tolerance at each recursive shuffling step was then used to spot additionally improved strains. Three highly tolerant strains were finally isolated and able to withstand up to 50 g/L isopropanol on plates. Even if increased tolerance to the desired end product was not always accompanied by higher production capabilities, some shuffled strains showed increased solvent titers compared to the parental strains and the original C. beijerinckii DSM 6423. This study confirms the efficiency of genome shuffling to generate improved strains toward a desired phenotype such as alcohol tolerance. This tool also offers the possibility of obtaining improved strains of Clostridium species for which targeted genetic engineering approaches have not been described yet.

  6. Attempts to identify Clostridium botulinum toxin in milk from three experimentally intoxicated Holstein cows

    Science.gov (United States)

    Moeller, R.B.; Puschner, B.; Walker, R.L.; Rocke, T.E.; Smith, S.R.; Cullor, J.S.; Ardans, A.A.

    2009-01-01

    Three adult lactating Holstein cows were injected in the subcutaneous abdominal vein with 175 ng/kg of body weight of Clostridium botulinum type C toxin (451 cow median toxic doses) to determine if this botulinum toxin crosses the blood–milk barrier. Whole blood (in sodium heparin) and clotted blood serum samples were taken at 0 min, 10 min, and 3, 6, 9, and 12 h postinoculation. Milk samples were taken at 0 min and at 3, 6, 9 and 12 h postinoculation. All samples were tested for the presence of the toxin using the mouse bioassay and immunostick ELISA test. The immunostick ELISA identified the toxin in whole blood and the mouse bioassay identified the toxin in serum at all times examined in all 3 animals. Toxin was not identified by either detection method in milk samples collected from the 3 animals. From these results, it appears that Clostridium botulinum type C toxin does not cross from the blood to the milk in detectable concentrations.

  7. Genome and Transcriptome of Clostridium phytofermentans, Catalyst for the Direct Conversion of Plant Feedstocks to Fuels.

    Directory of Open Access Journals (Sweden)

    Elsa Petit

    Full Text Available Clostridium phytofermentans was isolated from forest soil and is distinguished by its capacity to directly ferment plant cell wall polysaccharides into ethanol as the primary product, suggesting that it possesses unusual catabolic pathways. The objective of the present study was to understand the molecular mechanisms of biomass conversion to ethanol in a single organism, Clostridium phytofermentans, by analyzing its complete genome and transcriptome during growth on plant carbohydrates. The saccharolytic versatility of C. phytofermentans is reflected in a diversity of genes encoding ATP-binding cassette sugar transporters and glycoside hydrolases, many of which may have been acquired through horizontal gene transfer. These genes are frequently organized as operons that may be controlled individually by the many transcriptional regulators identified in the genome. Preferential ethanol production may be due to high levels of expression of multiple ethanol dehydrogenases and additional pathways maximizing ethanol yield. The genome also encodes three different proteinaceous bacterial microcompartments with the capacity to compartmentalize pathways that divert fermentation intermediates to various products. These characteristics make C. phytofermentans an attractive resource for improving the efficiency and speed of biomass conversion to biofuels.

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

    Science.gov (United States)

    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. © 2014 The Society for Applied Microbiology.

  9. The Incidence of Nosocomial Toxigenic Clostridium difficile Associated Diarrhea in Tehran Tertiary Medical Centers

    Directory of Open Access Journals (Sweden)

    Norakhoda Sadeghifard

    2010-10-01

    Full Text Available Clostridium difficile is the most common cause of nosocomial diarrhea. It is usually a consequence of antibiotic treatment, But sporadic cases can occur. This study was aimed to determine the frequency of the nosocomial Clostridium difficile (C. difficile associated diarrhea in Tehran University of Medical Sciences hospitals and study of antibacterial susceptibility of isolates. In this study a total of 942 stool samples from patients with nosocomial diarrhea that were hospitalized in Imam Khomeini hospital, Shariati hospital and Children clinical center were collected. The samples were cultured on a selective cycloserine cefoxitin fructose agar (CCFA and incubated in anaerobic conditions, at 37°C for 5 days. Isolates were characterized to species level by conventional biochemical tests. Bacterial cytotoxicity was assayed on tissue culture (vero. Antimicrobial sensitivity of isolated toxigenic C. difficile were investigated by kirby Beuer method (disk diffusion. Our findings show that, of the total patients, 57 toxigenic C. difficile (6.1% were isolated. Results of statistical analysis show significant differences between the rate of isolated toxigenic C. difficile and age group of patients (P

  10. Arylsulfotransferase from Clostridium innocuum-A new enzyme catalyst for sulfation of phenol-containing compounds.

    Science.gov (United States)

    Mozhaev, Vadim V; Khmelnitsky, Yuri L; Sanchez-Riera, Fernando; Maurina-Brunker, Julie; Rosson, Reinhardt A; Grund, Alan D

    2002-06-05

    Arylsulfotransferase (AST, EC 2.8.2.22), an enzyme capable of sulfating a wide range of phenol-containing compounds was purified from a Clostridium innocuum isolate (strain 554). The enzyme has a molecular weight of 320 kDa and is composed of four subunits. Unlike many mammalian and plant arylsulfotransferases, AST from Clostridium utilizes arylsulfates, including p-nitrophenyl sulfate, as sulfate donors, and is not reactive with 3-phosphoadenosine-5'-phosphosulfate (PAPS). The enzyme possesses broad substrate specificity and is active with a variety of phenols, quinones and flavonoids, but does not utilize primary and secondary alcohols and sugars as substrates. Arylsulfotransferase tolerates the presence of 10 vol% of polar cosolvents (dimethyl formamide, acetonitrile, methanol), but loses significant activity at higher solvent concentrations of 30-40 vol%. The enzyme retains high arylsulfotransferase activity in biphasic systems composed of water and nonpolar solvents, such as cyclohexane, toluene and chloroform, while in biphasic systems with more polar solvents (ethyl acetate, 2-pentanone, methyl tert-butyl ether, and butyl acetate) the enzyme activity is completely lost. High yields of AST-catalyzed sulfation were achieved in reactions with several phenols and tyrosine-containing peptides. Overall, AST studied in this work is a promising biocatalyst in organic synthesis to afford efficient sulfation of phenolic compounds under mild reaction conditions. Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 567-575, 2002.

  11. Fidaxomicin in the treatment of colitis due to Clostridium difficile: preliminary results

    Directory of Open Access Journals (Sweden)

    Francesco Cortese

    2014-12-01

    Full Text Available The incidence of Clostridium difficile infections (CDI and Clostridium difficile-Associated Diarrhea (CDAD is increasing in Canada, USA, and Europe and represents a considerable clinical problem. Both naïve and hypervirulent strains can be considered as opportunistic bacteria affecting immunocompromised, antibiotic-treated, critical, or subcritical patients with a microbiota disruption. CDI arising is strictly related to antibiotic, single or combined, and/or proton pump inhibitor treatment. CDI can cause a syndrome with systemic involvement and complex treatment, sometimes requiring surgical interventions (e.g. colectomy in fulminant colitis. Antibiotic treatment with metronidazole by mouth is the first choice and generally vancomycin is administered in case of lack of effectiveness. Fidaxomicin is a new macrocyclic antibiotic for C. difficile with microflora-sparing properties. This paper reports our initial experience in 11 patients with non-responder or relapsing CDIs. Fidaxomicin was effective in 10 cases (91%. Only one patient with an active ulcerative colitis did not respond and was treated with fecal-microbiota transplantation. In two patients diarrhea persisted, but just the ulcerative colitis one was C. difficile-related. No adverse events were experienced.http://dx.doi.org/10.7175/cmi.v8i1s.956

  12. Therapeutic Success of Rifaximin for Clostridium difficile Infection Refractory to Metronidazole and Vancomycin

    Directory of Open Access Journals (Sweden)

    George Tannous

    2010-09-01

    Full Text Available We report the case of a 46-year-old white male with confirmed Clostridium difficile infection for >4 weeks after fluoroquinolone therapy. The patient received two courses of metronidazole 500 mg three times daily (t.i.d. during which time diarrhea resolved; however, symptoms recurred 14–15 days after treatment termination. He received a 2-week course of vancomycin 125 mg four times daily, with symptoms recurring 10 days after treatment conclusion. The patient then received a pulsed tapering schedule of vancomycin with adjunctive Saccharomyces boulardii. Diarrhea recurred 12 days after treatment completion. He received rifaximin 400 mg t.i.d. while hospitalized for diarrhea-associated complications. Symptoms resolved within 24 h. The patient received a 4-week regimen of rifaximin 400 mg orally t.i.d. after discharge. No further episodes of diarrhea were reported within 6 months after treatment termination. The present case supports the potential benefit of rifaximin for the treatment of recurrent Clostridium difficile infection.

  13. Efficient dark fermentative hydrogen production from enzyme hydrolyzed rice straw by Clostridium pasteurianum (MTCC116).

    Science.gov (United States)

    Srivastava, Neha; Srivastava, Manish; Kushwaha, Deepika; Gupta, Vijai Kumar; Manikanta, Ambepu; Ramteke, P W; Mishra, P K

    2017-08-01

    In the present work, production of hydrogen via dark fermentation has been carried out using the hydrolyzed rice straw and Clostridium pasteurianum (MTCC116). The hydrolysis reaction of 1.0% alkali pretreated rice straw was performed at 70°C and 10% substrate loading via Fe 3 O 4 /Alginate nanocomposite (Fe 3 O 4 /Alginate NCs) treated thermostable crude cellulase enzyme following the previously established method. It is noticed that under the optimized conditions, at 70°C the Fe 3 O 4 /Alginate NCs treated cellulase has produced around 54.18g/L sugars as the rice straw hydrolyzate. Moreover, the efficiency of the process illustrates that using this hydrolyzate, Clostridium pasteurianum (MTCC116) could produce cumulative hydrogen of 2580ml/L in 144h with the maximum production rate of 23.96ml/L/h in 96h. In addition, maximum dry bacterial biomass of 1.02g/L and 1.51g/L was recorded after 96h and 144h, respectively with corresponding initial pH of 6.6 and 3.8, suggesting higher hydrogen production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Clostridium difficile Infections: A Global Overview of Drug Sensitivity and Resistance Mechanisms

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    Saeed S. Banawas

    2018-01-01

    Full Text Available Clostridium difficile (C. difficile is the most prevalent causative pathogen of healthcare-associated diarrhea. Notably, over the past 10 years, the number of Clostridium difficile outbreaks has increased with the rate of morbidity and mortality. The occurrence and spread of C. difficile strains that are resistant to multiple antimicrobial drugs complicate prevention as well as potential treatment options. Most C. difficile isolates are still susceptible to metronidazole and vancomycin. Incidences of C. difficile resistance to other antimicrobial drugs have also been reported. Most of the antibiotics correlated with C. difficile infection (CDI, such as ampicillin, amoxicillin, cephalosporins, clindamycin, and fluoroquinolones, continue to be associated with the highest risk for CDI. Still, the detailed mechanism of resistance to metronidazole or vancomycin is not clear. Alternation in the target sites of the antibiotics is the main mechanism of erythromycin, fluoroquinolone, and rifamycin resistance in C. difficile. In this review, different antimicrobial agents are discussed and C. difficile resistance patterns and their mechanism of survival are summarized.

  15. TcdC does not significantly repress toxin expression in Clostridium difficile 630ΔErm.

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    Dennis Bakker

    Full Text Available In the past decade, Clostridium difficile has emerged as an important gut pathogen. Symptoms of C. difficile infection range from mild diarrhea to pseudomembranous colitis, sometimes resulting in colectomy or death. The main virulence factors of C. difficile are toxin A and toxin B. Besides the genes encoding these toxins (tcdA and tcdB, the pathogenicity locus (PaLoc also contains genes encoding a sigma factor (tcdR and a putative anti-sigma factor (tcdC. The important role of TcdR as a sigma factor for toxin expression is undisputed, whereas the role of TcdC as an anti-sigma factor, inhibiting toxin expression, is currently the subject of debate. To clarify the role of TcdC in toxin expression, we generated an isogenic ClosTron-based mutant of tcdC in Clostridium difficile strain 630Δ Erm (CT::tcdC and determined the transcription levels of the PaLoc genes and the expression levels of the toxins in the wild type strain and the tcdC mutant strain. We found only minor differences in transcription levels of the PaLoc genes between the wild type and CT::tcdC strains and total toxin levels did not significantly differ either. These results suggest that in C. difficile 630Δerm TcdC is not a major regulator of toxin expression under the conditions tested.

  16. Clinical manifestations of Clostridium difficile infection in a medical center in Taiwan.

    Science.gov (United States)

    Lai, Chih-Cheng; Lin, Sheng-Hsiang; Tan, Che-Kim; Liao, Chun-Hsing; Huang, Yu-Tsung; Hsueh, Po-Ren

    2014-12-01

    To investigate the clinical characteristics of Clostridium difficile infection (CDI) at a medical center in Taiwan. Patients with CDI were identified from medical records at the National Taiwan University Hospital (Taipei, Taiwan). The following information was gathered and analyzed to better understand the clinical manifestations of CDI: age; sex; underlying immunocompromised conditions; laboratory data; in-hospital mortality; and previous use of drugs such as antimicrobial agents, steroids, and antipeptic ulcer agents. During the years 2000-2010, 122 patients were identified as having CDI. This included 92 patients with nontoxigenic CDI (i.e., positive stool culture for C. difficile but negative results for toxins A and B) and 30 patients with toxigenic CDI (i.e., positive stool culture cultures for C. difficile and positive results for toxins A and B). Of the 122 patients, 48 (39%) patients were older than 65 years and most patients acquired the CDI while in the hospital. Active cancer was the most common reason for hospitalization, followed by diabetes mellitus, and end-stage renal disease. More than 90% of the patients had received antibiotics before acquiring CDI. The results of fecal leukocyte examinations were positive in 33 (27%) patients. The overall in-hospital mortality rate was 26.2%. There were no significant differences between patients with nontoxigenic CDI and patients with toxigenic CDI. Clostridium difficile infection can develop in healthcare facilities and in community settings, especially in immunocompromised patients. Copyright © 2013. Published by Elsevier B.V.

  17. Impact of end stage kidney disease on costs and outcomes of Clostridium difficile infection.

    Science.gov (United States)

    Goyal, Abhinav; Chatterjee, Kshitij; Yadlapati, Sujani; Rangaswami, Janani

    2017-09-01

    To assess the impact of end stage kidney disease (ESKD) on the outcomes of Clostridium difficile infection (CDI), including complications of infection, length of hospital stay, overall mortality, and healthcare burden. The National Inpatient Sample (NIS) database created by the Agency of Healthcare Research and Quality (AHRQ) was used, covering the years 2009 through 2013. Manufacturer-provided sampling weights were used to produce national estimates. All-cause unadjusted in-hospital mortality was significantly higher for patients with CDI and ESKD than for patients without ESKD (11.6% vs. 7.7%, pcost of hospitalization for patients with CDI and ESKD was also significantly higher compared to the non-ESKD group (USD $35 588 vs. $23 505, in terms of the 2013 value of the USD, pClostridium difficile infection is associated with higher mortality, a longer length of stay, and a higher cost of hospitalization. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  18. Treatment of pediatric Clostridium difficile infection: a review on treatment efficacy and economic value

    Directory of Open Access Journals (Sweden)

    D'Ostroph AR

    2017-10-01

    Full Text Available Amanda R D’Ostroph,1 Tsz-Yin So2 1UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, 2Department of Pharmacy, Moses H Cone Memorial Hospital, Greensboro, NC, USA Abstract: The incidence of Clostridium difficile infection (CDI in pediatric patients continues to rise. Most of the pediatric recommendations for CDI treatment are extrapolated from the literature and guidelines for adults. The American Academy of Pediatrics recommends oral metronidazole as the first-line treatment option for an initial CDI and the first recurrence if they are mild to moderate in severity. Oral vancomycin is recommended to be used for severe CDI and the second recurrent infection. Additional pulsed regimen of oral vancomycin, which is tapered, may increase efficacy in refractory patients. However, there is lack of large studies evaluating the use of fidaxomicin in pediatrics to know whether it could be a safe and effective treatment option for difficult-to-treat patients. Fidaxomicin is associated with higher total drug costs compared to metronidazole and vancomycin, but the literature supports its use due to a lower rate of CDI recurrence, which may result in cost savings. Further studies are warranted to evaluate the use of fidaxomicin in patients <18 years old and to understand its role in the standard of care for pediatric patients with CDI. Keywords: Clostridium difficile, diarrhea, fidaxomicin, vancomycin, metronidazole, pediatrics 

  19. Hospitalization stay and costs attributable to Clostridium difficile infection: a critical review.

    Science.gov (United States)

    Gabriel, L; Beriot-Mathiot, A

    2014-09-01

    In most healthcare systems, third-party payers fund the costs for patients admitted to hospital for Clostridium difficile infection (CDI) whereas, for CDI cases arising as complications of hospitalization, not all related costs are refundable to the hospital. We therefore aimed to critically review and categorize hospital costs and length of hospital stay (LOS) attributable to Clostridium difficile infection and to investigate the economic burden associated with it. A comprehensive literature review selected papers describing the costs and LOS for hospitalized patients as outcomes of CDI, following the use of statistics to identify costs and LOS solely attributable to CDI. Twenty-four studies were selected. Estimated attributable costs, all ranges expressed in US dollars, were $6,774-$10,212 for CDI requiring admission, $2,992-$29,000 for hospital-acquired CDI, and $2,454-$12,850 where no categorization was made. The ranges for LOS values were 5-13.6, 2.7-21.3, and 2.8-17.9 days, respectively. The categorization of CDI attributable costs allows budget holders to anticipate the cost per CDI case, a perspective that should enrich the design of appropriate incentives for the various budget holders to invest in prevention so that CDI prevention is optimized globally. Copyright © 2014 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

  20. Cadaver Thanatomicrobiome Signatures: The Ubiquitous Nature of Clostridium Species in Human Decomposition

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    Gulnaz T. Javan

    2017-10-01

    Full Text Available Human thanatomicrobiome studies have established that an abundant number of putrefactive bacteria within internal organs of decaying bodies are obligate anaerobes, Clostridium spp. These microorganisms have been implicated as etiological agents in potentially life-threatening infections; notwithstanding, the scale and trajectory of these microbes after death have not been elucidated. We performed phylogenetic surveys of thanatomicrobiome signatures of cadavers’ internal organs to compare the microbial diversity between the 16S rRNA gene V4 hypervariable region and V3-4 conjoined regions from livers and spleens of 45 cadavers undergoing forensic microbiological studies. Phylogenetic analyses of 16S rRNA gene sequences revealed that the V4 region had a significantly higher mean Chao1 richness within the total microbiome data. Permutational multivariate analysis of variance statistical tests, based on unweighted UniFrac distances, demonstrated that taxa compositions were significantly different between V4 and V3-4 hypervariable regions (p < 0.001. Of note, we present the first study, using the largest cohort of criminal cases to date, that two hypervariable regions show discriminatory power for human postmortem microbial diversity. In conclusion, here we propose the impact of hypervariable region selection for the 16S rRNA gene in differentiating thanatomicrobiomic profiles to provide empirical data to explain a unique concept, the Postmortem Clostridium Effect.

  1. Retargeting the Clostridium botulinum C2 toxin to the neuronal cytosol.

    Science.gov (United States)

    Pavlik, Benjamin J; Hruska, Elizabeth J; Van Cott, Kevin E; Blum, Paul H

    2016-03-30

    Many biological toxins are known to attack specific cell types, delivering their enzymatic payloads to the cytosol. This process can be manipulated by molecular engineering of chimeric toxins. Using toxins with naturally unlinked components as a starting point is advantageous because it allows for the development of payloads separately from the binding/translocation components. Here the Clostridium botulinum C2 binding/translocation domain was retargeted to neural cell populations by deleting its non-specific binding domain and replacing it with a C. botulinum neurotoxin binding domain. This fusion protein was used to deliver fluorescently labeled payloads to Neuro-2a cells. Intracellular delivery was quantified by flow cytometry and found to be dependent on artificial enrichment of cells with the polysialoganglioside receptor GT1b. Visualization by confocal microscopy showed a dissociation of payloads from the early endosome indicating translocation of the chimeric toxin. The natural Clostridium botulinum C2 toxin was then delivered to human glioblastoma A172 and synchronized HeLa cells. In the presence of the fusion protein, native cytosolic enzymatic activity of the enzyme was observed and found to be GT1b-dependent. This retargeted toxin may enable delivery of therapeutics to peripheral neurons and be of use in addressing experimental questions about neural physiology.

  2. Binding properties of Clostridium botulinum type C progenitor toxin to mucins.

    Science.gov (United States)

    Nakamura, Toshio; Takada, Noriko; Tonozuka, Takashi; Sakano, Yoshiyuki; Oguma, Keiji; Nishikawa, Atsushi

    2007-04-01

    It has been reported that Clostridium botulinum type C 16S progenitor toxin (C16S toxin) first binds to the sialic acid on the cell surface of mucin before invading cells [A. Nishikawa, N. Uotsu, H. Arimitsu, J.C. Lee, Y. Miura, Y. Fujinaga, H. Nakada, T. Watanabe, T. Ohyama, Y. Sakano, K. Oguma, The receptor and transporter for internalization of Clostridium botulinum type C progenitor toxin into HT-29 cells, Biochem. Biophys. Res. Commun. 319 (2004) 327-333]. In this study we investigated the binding properties of the C16S toxin to glycoproteins. Although the toxin bound to membrane blotted mucin derived from the bovine submaxillary gland (BSM), which contains a lot of sialyl oligosaccharides, it did not bind to neuraminidase-treated BSM. The binding of the toxin to BSM was inhibited by N-acetylneuraminic acid, N-glycolylneuraminic acid, and sialyl oligosaccharides strongly, but was not inhibited by neutral oligosaccharides. Both sialyl alpha2-3 lactose and sialyl alpha2-6 lactose prevented binding similarly. On the other hand, the toxin also bound well to porcine gastric mucin. In this case, neutral oligosaccharides might play an important role as ligand, since galactose and lactose inhibited binding. These results suggest that the toxin is capable of recognizing a wide variety of oligosaccharide structures.

  3. Clostridium difficile from food and surface samples in a Belgian nursing home: an unlikely source of contamination.

    Science.gov (United States)

    Rodriguez, C; Korsak, N; Taminiau, B; Avesani, V; Van Broeck, J; Brach, P; Delmée, M; Daube, G

    2015-04-01

    This study investigates the contamination of foods and surfaces with Clostridium difficile in a single nursing home. C. difficile PCR-ribotype 078 was found in one food sample and in none of the tested surfaces. These results indicate that food and surfaces are an unlikely source of C. difficile infection in this setting. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. First Insights into the Genome Sequence of Clostridium thermopalmarium DSM 5974, a Butyrate-Producing Bacterium Isolated from Palm Wine.

    Science.gov (United States)

    Poehlein, Anja; Hettwer, Eva; Mohnike, Lennart; Daniel, Rolf

    2018-04-26

    Clostridium thermopalmarium is a moderate thermophilic, rod-shaped, and endospore-forming bacterium, which was isolated from palm wine in Senegal. Butyrate is produced from a broad variety of sugar substrates. Here, we present the draft genome sequence of C. thermopalmarium DSM 5974 (2.822 Mb) containing 2,665 predicted protein-encoding genes. Copyright © 2018 Poehlein et al.

  5. Syntrophic co-culture of aerobic Bacillus and anaerobic Clostridium for bio-fuels and bio-hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Jui-Jen; Ho, Cheng-Yu.; Chen, Wei-En; Huang, Chieh-Chen [Department of Life Sciences, National Chung Hsing University, Taichung (China); Chou, Chia-Hung; Lay, Jiunn-Jyi [Department of Science and Technology, National Kaohsiung First University, Kaohsiung (China)

    2008-10-15

    By using brewery yeast waste and microflora from rice straw compost, an anaerobic semi-solid bio-hydrogen-producing system has been established. For the purpose of industrialization, the major players of both aerobic and anaerobic bacterial strains in the system were isolated and their combination for an effective production of bio-hydrogen and other bio-fuels was examined in this study. The phylogenetic analysis found that four anaerobic isolates (Clostridium beijerinckii L9, Clostridium diolis Z2, Clostridium roseum Z5-1, and C. roseum W8) were highly related with each other and belongs to the cluster I clostridia family, the family that many of solvent-producing strains included. On the other hand, one of the aerobic isolates, the Bacillus thermoamylovorans strain I, shown multiple extracellular enzyme activities including lipase, protease, {alpha}-amylase, pectinase and cellulase, was suggested as a good partner for creating an anaerobic environment and pre-saccharification of substrate for those co-cultured solventogenic clostridial strain. Among these clostridial strains, though C. beijerinckii L9 do not show as many extracellular enzyme activities as Bacillus, but it performs the highest hydrogen-producing ability. The original microflora can be updated to a syntrophic bacterial co-culture system contended only with B. thermoamylovorans I and C. beijerinckii L9. The combination of aerobic Bacillus and anaerobic Clostridium may play the key role for developing the industrialized bio-fuels and bio-hydrogen-producing system from biomass. (author)

  6. Analysing the dhaT gene in Colombian Clostridium sp. (Clostridia 1,3-propanediol-producing strains

    Directory of Open Access Journals (Sweden)

    Diana Milena Quilaguy-Ayure

    2010-04-01

    Full Text Available To analyze the dhaT gene, one of the genes responsible for the 1,3-propanediol (1,3-PD production, in two native Clostridiumstrains. Materials and methods: The dhaT gene was amplified by Polimerase Chain Reaction with specific primers designed fromClostridium butyricum VPI1718 operon. Bioinformatics tools like BLASTN, ORF finder, BLASTP and ClustalW were used to determinethe identity of the sequence and to assign a function. Results: DNA amplification products were obtained from Colombian Clostridium sp.native strains (IBUN 13A and IBUN 158B and the Clostridium butyricum DSM 2478 strain, which were sequenced. According to thebioinformatics analysis of the above sequences, a high degree of similarity was found with the dhaT gene of different bacterial species. Thehighest percentage of identity was obtained with the Clostridium butyricum VPI 1718 strain. Conclusion: knowledge of the physicalstructure of the 1,3-PD operon in native strains opens the way for developing genetic and metabolic engineering strategies for improvingprocesses productivity.

  7. Different Roles Of Class-I And Class-II Clostridium-histolyticum Collagenase In Rat Pancreatic-islet Isolation

    NARCIS (Netherlands)

    Wolters, G. H. J.; Vos-Scheperkeuter, Greetje; Lin, Hun-Chi; van Schilfaarde, R

    Crude Clostridium histolyticum collagenase was purified by gel filtration and fractionated by anion exchange chromatography into class I with high collagen digestion activity (CDA) and low FALGPA (2-furanacryloyl-L-leucylglycyl-L-prolyI-L-alanine )hydrolysis activity (FHA), class II with low CDA and

  8. The mycotoxin deoxynivalenol predisposes for the development of Clostridium perfringens-induced necrotic enteritis in broiler chickens.

    Science.gov (United States)

    Antonissen, Gunther; Van Immerseel, Filip; Pasmans, Frank; 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% (Peffect 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.

  9. Comparison of Culture, Cytotoxin Assay and Two Eia Tests with Clinical Diagnosis of Clostridium difficile-Associated Diarrhea

    Directory of Open Access Journals (Sweden)

    Marilyn Binning

    1994-01-01

    Full Text Available Objective: The most common etiology of infectious diarrhea in hospitalized patients is Clostridium difficile. No single laboratory test yields a definitive diagnosis. Four methods were evaluated for their sensitivity and specificity in patients who had clinically defined C difficile-associated diarrhea.

  10. Modification of an acetone-sodium dodecyl sulfate disruption method for cellular protein extraction from neuropathogenic Clostridium botulinum

    Science.gov (United States)

    An acetone-sodium dodecyl sulfate (SDS) disruption method was used for the extraction of cellular proteins from neurotoxigenic Clostridium botulinum. The amount of protein extracted per gram of dry weight and the protein profile as revealed by polyacrylamide gel electrophoresis (PAGE) was comparabl...

  11. Clostridium botulinum group I strain genotyping by 15-locus multilocus variable-number tandem-repeat analysis

    NARCIS (Netherlands)

    Fillo, S.; Giordani, F.; Anniballi, F.; Gorgé, O.; Ramisse, V.; Vergnaud, G.; Riehm, J.M.; Scholz, H.C.; Splettstoesser, W.D.; Kieboom, J.; Olsen, J.-S.; Fenicia, L.; Lista, F.

    2011-01-01

    Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of

  12. A Multi-Center Prospective Derivation and Validation of a Clinical Prediction Tool for Severe Clostridium difficile Infection.

    LENUS (Irish Health Repository)

    Na, Xi

    2015-04-23

    Prediction of severe clinical outcomes in Clostridium difficile infection (CDI) is important to inform management decisions for optimum patient care. Currently, treatment recommendations for CDI vary based on disease severity but validated methods to predict severe disease are lacking. The aim of the study was to derive and validate a clinical prediction tool for severe outcomes in CDI.

  13. All-cause and disease-specific mortality in hospitalized patients with Clostridium difficile infection: a multicenter cohort study

    NARCIS (Netherlands)

    Hensgens, Marjolein P. M.; Goorhuis, Abraham; Dekkers, Olaf M.; van Benthem, Birgit H. B.; Kuijper, Ed J.

    2013-01-01

    Mortality among patients with Clostridium difficile infection (CDI) is high. Because of high age and multiple underlying diseases, CDI-related mortality is difficult to estimate. We estimated CDI-related mortality in an endemic situation, not influenced by outbreaks and consequently certain patients

  14. In situ detection of the Clostridium botulinum type C1 toxin gene in wetland sediments with a nested PCR assay

    Science.gov (United States)

    Williamson, Judy L.; Rocke, Tonie E.; Aiken, Judd M.

    1999-01-01

    A nested PCR was developed for detection of the Clostridium botulinum type C1 toxin gene in sediments collected from wetlands where avian botulism outbreaks had or had not occurred. The C1 toxin gene was detected in 16 of 18 sites, demonstrating both the ubiquitous distribution of C. botulinum type C in wetland sediments and the sensitivity of the detection assay.

  15. Elucidating the contributions of multiple aldehyde/alcohol dehydrogenases to butanol and ethanol production in Clostridium acetobutylicum

    OpenAIRE

    Dai, Zongjie; Dong, Hongjun; Zhang, Yanping; Li, Yin

    2016-01-01

    Ethanol and butanol biosynthesis in Clostridium acetobutylicum share common aldehyde/alcohol dehydrogenases. However, little is known about the relative contributions of these multiple dehydrogenases to ethanol and butanol production respectively. The contributions of six aldehyde/alcohol dehydrogenases of C. acetobutylicum on butanol and ethanol production were evaluated through inactivation of the corresponding genes respectively. For butanol production, the relative contributions from thes...

  16. Whole-genome sequence of Clostridium lituseburense L74, isolated from the larval gut of the rhinoceros beetle, Trypoxylus dichotomus.

    Science.gov (United States)

    Lee, Yookyung; Lim, Sooyeon; Rhee, Moon-Soo; Chang, Dong-Ho; Kim, Byoung-Chan

    2016-03-01

    Clostridium lituseburense L74 was isolated from the larval gut of the rhinoceros beetle, Trypoxylus dichotomus collected in Yeong-dong, Chuncheongbuk-do, South Korea and subjected to whole genome sequencing on HiSeq platform and annotated on RAST. The nucleotide sequence of this genome was deposited into DDBJ/EMBL/GenBank under the accession NZ_LITJ00000000.

  17. Current challenges in the treatment of severe Clostridium difficile infection: early treatment potential of fecal microbiota transplantation

    NARCIS (Netherlands)

    van Beurden, Yvette H.; Nieuwdorp, Max; van de Berg, Pablo J. E. J.; Mulder, Chris J. J.; Goorhuis, Abraham

    2017-01-01

    Fecal microbiota transplantation (FMT) is a very effective treatment for recurrent Clostridium difficile infection (CDI). Less is known about the application of FMT as a curative treatment of severe or complicated CDI. In this review, we present and discuss evidence supporting the curative use of

  18. European Society of Clinical Microbiology and Infectious Diseases: update of the treatment guidance document for Clostridium difficile infection

    NARCIS (Netherlands)

    Debast, S.B.; Bauer, M.P.; Kuijper, E.J.; et al.,

    2014-01-01

    In 2009 the first European Society of Clinical Microbiology and Infection (ESCMID) treatment guidance document for Clostridium difficile infection (CDI) was published. The guideline has been applied widely in clinical practice. In this document an update and review on the comparative effectiveness

  19. Mutant form C115H of Clostridium sporogenes methionine γ-lyase efficiently cleaves S-Alk(en)yl-l-cysteine sulfoxides to antibacterial thiosulfinates.

    Science.gov (United States)

    Kulikova, Vitalia V; Anufrieva, Natalya V; Revtovich, Svetlana V; Chernov, Alexander S; Telegin, Georgii B; Morozova, Elena A; Demidkina, Tatyana V

    2016-10-01

    Pyridoxal 5'-phosphate-dependent methionine γ-lyase (MGL) catalyzes the β-elimination reaction of S-alk(en)yl-l-cysteine sulfoxides to thiosulfinates, which possess antimicrobial activity. Partial inactivation of the enzyme in the course of the reaction occurs due to oxidation of active site cysteine 115 conserved in bacterial MGLs. In this work, the C115H mutant form of Clostridium sporogenes MGL was prepared and the steady-state kinetic parameters of the enzyme were determined. The substitution results in an increase in the catalytic efficiency of the mutant form towards S-substituted l-cysteine sulfoxides compared to the wild type enzyme. We used a sulfoxide/enzyme system to generate antibacterial activity in situ. Two-component systems composed of the mutant enzyme and three S-substituted l-cysteine sulfoxides were demonstrated to be effective against Gram-positive and Gram-negative bacteria and three clinical isolates from mice. © 2016 IUBMB Life, 68(10):830-835, 2016. © 2016 International Union of Biochemistry and Molecular Biology.

  20. Influence of MAP and Multi-layer Flexible Pouches on Clostridium Count of Smoked Kutum Fish (Rutilus frisii kutum

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    Nazanin Zand

    2016-11-01

    Full Text Available In this study the effect of different concentrations of three gas mixture (carbon dioxide, nitrogen, oxygen, and also vacuum conditions and flexible multi-layer films were evaluated on Clostridium count of smoked kutum fish (Rutilus frisii kutum at ambient condition (T= 25 0C. Ordinary condition as control packaging were compared with four types of modified atmosphere packaging: (N270%+ CO230%, (N230% + CO270%, (45%CO2+ 45%N2+10%O2 and vacuum conditions, in this project. Smoked kutum fish were packaged into 3 kinds of flexible pouches {3- layers(PET(12/AL(12/LLD(100, 4-layers (PET(12/AL(7/ PET(12/LLD(100, and 3-layer (PET(12/AL(7/LLD(100}. Packed samples were performed microbial tests (Clostridium count, in different times during 60 days, with 15 treatment ,3 run, statistical analysis and comparison of data, were done by software SAS (Ver:9/1 and Duncan’s new multiple range test, with confidence level of 95% (P <0.05 . The shelf life of Samples (according to Clostridium count were reported in 4-layers , under conditions 1,2,3 and vacuum conditions, 60,58,45,40 days, in 3-layers (AL:12, under conditions 1,2,3 were 55,50,40 days and in vacuum conditions were about 35 days, with 3- layers(AL:7, under conditions 1,2,3 and vacuum conditions 45,40,35, 30 days. Clostridium count showed that increasing CO2 concentration prolonged shelf life. During the period of this experiment Clostridium count of samples in various conditions, had significant level. According to these results could be concluded the best condition belonged to treatment under modified atmosphere CO2 70% and also 4- layer container due to the thickness (131 μ, low permeability of water vapor in this 4-layer container and anti-microbial effect of more percentage of CO2.