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1

The bc:caa3 supercomplexes from the Gram positive bacterium Bacillus subtilis respiratory chain: a megacomplex organization?  

PubMed

The respiratory chain of some prokaryotes was shown to be organized in supercomplexes. This association has been proposed to improve enzyme stability and the overall efficiency of the oxidative phosphorylation process. Here, we have revisited recent data on the supercomplexes of Bacillus subtilis respiratory chain, by means of 1D and 2D-BN-PAGE, sucrose gradient fractionation of solubilized membranes, and mass spectrometry analysis of BN-PAGE bands detected in gel for succinate and cytochrome c oxidoreductase activities. The cytochrome bc:caa3 oxygen oxidoreductase supercomplex was observed in different stoichiometries, (bc)4:(caa3)2, (bc)2:(caa3)4 and 2[(bc)2:(caa3)4], suggesting for the first time the string association model of supercomplexes in a Gram positive bacterium. In addition, the presence of a succinate:quinone oxidoreductase:nitrate reductase supercomplex was confirmed by the co-localized succinate:nitroblue tetrazolium and methylviologen:nitrate oxidoreductase activities detected in gel and corroborated by LC-MS/MS analysis. PMID:23880299

Sousa, Pedro M F; Videira, Marco A M; Santos, Filipe A S; Hood, Brian L; Conrads, Thomas P; Melo, Ana M P

2013-09-01

2

Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans  

NASA Technical Reports Server (NTRS)

A rapid, sensitive, inexpensive in situ hybridization technique, using 30-mer 16S rRNA probes, can specifically differentiate two closely related Bacillus spp., B. polymyxa and B. macerans. The 16S rRNA probes were labeled with a rhodamine derivative (Texas Red), and quantitative fluorescence measurements were made on individual bacterial cells. The microscopic fields analyzed were selected by phase-contrast microscopy, and the fluorescence imaging analyses were performed on 16 to 67 individual cells. The labeled 16S rRNA probe, POL, whose sequence was a 100% match with B. polymyxa 16S rRNA but only a 60% match with B. macerans 16S rRNA, gave quantitative fluorescence ratio measurements that were 34.8-fold higher for B. polymyxa cells than for B. macerans cells. Conversely, the labeled probe, MAC, which matched B. polymyxa 16S rRNA in 86.6% of its positions and B. macerans 16S rRNA in 100% of its positions, gave quantitative fluorescence measurements that were 59.3-fold higher in B. macerans cells than in B. polymyxa cells. Control probes, whose 16S rRNA sequence segment (P-M) was present in both B. polymyxa and B. macerans as well as a panprokaryotic probe (16S), having a 100% match with all known bacteria, hybridized equally well with both organisms. These latter hybridizations generated very high fluorescence signals, but their comparative fluorescence ratios (the differences between two organisms) were low. The control paneukaryotic probe (28S), which had less than 30% identity for both B. macerans and B. polymyxa, did not hybridize with either organism.

Jurtshuk, R. J.; Blick, M.; Bresser, J.; Fox, G. E.; Jurtshuk, P. Jr

1992-01-01

3

The acetylproteome of Gram-positive model bacterium Bacillus subtilis.  

PubMed

N(?) -lysine acetylation, a reversible and highly regulated PTM, has been shown to occur in the model Gram-negative bacteria Escherichia coli and Salmonella enterica. Here, we extend this acetylproteome analysis to Bacillus subtilis, a model Gram-positive bacterium. Through anti-acetyllysine antibody-based immunoseparation of acetylpeptides followed by nano-HPLC/MS/MS analysis, we identified 332 unique lysine-acetylated sites on 185 proteins. These proteins are mainly involved in cellular housekeeping functions such as central metabolism and protein synthesis. Fifity-nine of the lysine-acetylated proteins showed homology with lysine-acetylated proteins previously identified in E. coli, suggesting that acetylated proteins are more conserved. Notably, acetylation was found at or near the active sites predicted by Prosite signature, including SdhA, RocA, Kbl, YwjH, and YfmT, indicating that lysine acetylation may affect their activities. In 2-amino-3-ketobutyrate CoA ligase Kbl, a class II aminotransferase, a lysine residue involved in pyridoxal phosphate attachment was found to be acetylated. This data set provides evidence for the generality of lysine acetylation in eubacteria and opens opportunities to explore the consequences of acetylation modification on the molecular physiology of B. subtilis. PMID:23468065

Kim, Dooil; Yu, Byung Jo; Kim, Jung Ae; Lee, Yong-Jik; Choi, Soo-Geun; Kang, Sunghyun; Pan, Jae-Gu

2013-05-01

4

Polyhydroxyalkanoates in Gram-positive bacteria: insights from the genera Bacillus and Streptomyces  

Microsoft Academic Search

Gram-positive bacteria, notably Bacillus and Streptomyces, have been used extensively in industry. However, these microorganisms have not yet been exploited for the production of the biodegradable polymers, polyhydroxyalkanoates (PHAs). Although PHAs have many potential applications, the cost of production means that medical applications are currently the main area of use. Gram-negative bacteria, currently the only commercial source of PHAs, have

Sabeel P. Valappil; Aldo R. Boccaccini; Christopher Bucke; Ipsita Roy

2007-01-01

5

The complete genome sequence of the Gram-positive bacterium Bacillus subtilis  

Microsoft Academic Search

Bacillus subtilis is the best-characterized member of the Gram-positive bacteria. Its genome of 4,214,810 base pairs comprises 4,100 protein-coding genes. Of these protein-coding genes, 53% are represented once, while a quarter of the genome corresponds to several gene families that have been greatly expanded by gene duplication, the largest family containing 77 putative ATP-binding transport proteins. In addition, a large

F. Kunst; N. Ogasawara; I. Moszer; A. M. Albertini; G. Alloni; V. Azevedo; M. G. Bertero; P. Bessières; A. Bolotin; S. Borchert; R. Borriss; L. Boursier; A. Brans; M. Braun; S. C. Brignell; S. Bron; S. Brouillet; C. V. Bruschi; B. Caldwell; V. Capuano; N. M. Carter; S.-K. Choi; J.-J. Codani; I. F. Connerton; N. J. Cummings; R. A. Daniel; F. Denizot; K. M. Devine; A. Düsterhöft; S. D. Ehrlich; P. T. Emmerson; K. D. Entian; J. Errington; C. Fabret; E. Ferrari; D. Foulger; C. Fritz; M. Fujita; Y. Fujita; S. Fuma; A. Galizzi; N. Galleron; S.-Y. Ghim; P. Glaser; A. Goffeau; E. J. Golightly; G. Grandi; G. Guiseppi; B. J. Guy; K. Haga; J. Haiech; C. R. Harwood; A. Hénaut; H. Hilbert; S. Holsappel; S. Hosono; M.-F. Hullo; M. Itaya; L. Jones; B. Joris; D. Karamata; Y. Kasahara; M. Klaerr-Blanchard; C. Klein; Y. Kobayashi; P. Koetter; G. Koningstein; S. Krogh; M. Kumano; K. Kurita; A. Lapidus; S. Lardinois; J. Lauber; V. Lazarevic; S.-M. Lee; A. Levine; H. Liu; S. Masuda; C. Mauël; C. Médigue; N. Medina; R. P. Mellado; M. Mizuno; D. Moestl; S. Nakai; M. Noback; D. Noone; M. O'Reilly; K. Ogawa; A. Ogiwara; B. Oudega; S.-H. Park; V. Parro; T. M. Pohl; D. Portetelle; S. Porwollik; A. M. Prescott; E. Presecan; P. Pujic; B. Purnelle; G. Rapoport; M. Rieger; S. Reynolds; C. Rivolta; E. Rocha; B. Roche; M. Rose; Y. Sadaie; T. Sato; E. Scanlan; S. Schleich; R. Schroeter; F. Scoffone; J. Sekiguchi; A. Sekowska; S. J. Seror; P. Serror; B.-S. Shin; B. Soldo; A. Sorokin; E. Tacconi; T. Takagi; H. Takahashi; K. Takemaru; M. Takeuchi; A. Tamakoshi; T. Tanaka; P. Terpstra; A. Tognoni; V. Tosato; S. Uchiyama; M. Vandenbol; F. Vannier; A. Vassarotti; A. Viari; R. Wambutt; E. Wedler; H. Wedler; T. Weitzenegger; P. Winters; A. Wipat; H. Yamamoto; K. Yamane; K. Yasumoto; K. Yata; K. Yoshida; H.-F. Yoshikawa; E. Zumstein; H. Yoshikawa; A. Danchin

1997-01-01

6

The complete genome sequence of the gram-positive bacterium Bacillus subtilis  

Microsoft Academic Search

Bacillus subtilis is the best-characterized member of the Gram-positive bacteria. Its genome of 4,214,810 base pairs comprises 4,100 protein-coding genes. Of these protein-coding genes, 53% are represented once, while a quarter of the genome corresponds to several gene families that have been greatly expanded by gene duplication, the largest family containing 77 putative ATP-binding transport proteins. In addition, a large

F. Kunst; N. Ogasawara; I. Moszer; A. M. Albertini; G. Alloni; V. Azevedo; M. G. Bertero; P. Bessières; A. Bolotin; S. Borchert; R. Borriss; L. Boursier; A. Brans; M. Braun; S. C. Brignell; S. Bron; S. Brouillet; C. V. Bruschi; B. Caldwell; V. Capuano; N. M. Carter; S.-K. Choi; J.-J. Codani; I. F. Connerton; A. Danchin

1997-01-01

7

Organization of genes for tetrapyrrole biosynthesis in gram--positive bacteria.  

PubMed

Clusters of genes encoding enzymes for tetrapyrrole biosynthesis were cloned from Bacillus sphaericus, Bacillus stearothermophilus, Brevibacillus brevis and Paenibacillus macerans. The sequences of all hemX genes found, and of a 6.3 kbp hem gene cluster from P. macerans, were determined. The structure of the hem gene clusters was compared to that of other Gram-positive bacteria. The Bacillus and Brevibacillus species have a conserved organization of the genes hemAXCDBL, required for biosynthesis of uroporphyrinogen III (UroIII) from glutamyl-tRNA. In P. macerans, the hem genes for UroIII synthesis are also closely linked but their organization is different: there is no hemX gene and the gene cluster also contains genes, cysG8 and cysG(A)-hemD, encoding the enzymes required for synthesis of sirohaem from UroIII. Bacillus subtilis contains genes for three proteins, NasF, YInD and YInF, with sequence similarity to Escherichia coli CysG, which is a multi-functional protein catalysing sirohaem synthesis from UroIII. It is shown that YInF is required for sirohaem synthesis and probably catalyses the precorrin-2 to sirohaem conversion. YInD probably catalyses precorrin-2 synthesis from UroIII and NasF seems to be specific for nitrite reduction. PMID:10217486

Johansson, P; Hederstedt, L

1999-03-01

8

Genetic determinants of antimicrobial resistance in Gram positive bacteria from organic foods.  

PubMed

Bacterial biocide resistance is becoming a matter of concern. In the present study, a collection of biocide-resistant, Gram-positive bacteria from organic foods (including 11 isolates from genus Bacillus, 25 from Enterococcus and 10 from Staphylococcus) were analyzed for genes associated to biocide resistance efflux pumps and antibiotic resistance. The only qac-genes detected were qacA/B (one Bacillus cereus isolate) and smr (one B. cereus and two Staphylococcus saprophyticus isolates). Efflux pump genes efrA and efrB genes were detected in Staphylococcus (60% of isolates), Bacillus (54.54%) and Enterococcus (24%); sugE was detected in Enterococcus (20%) and in one Bacillus licheniformis; mepA was detected in Staphylococcus (60%) and in one Enterococcus isolate (which also carried mdeA), and norE gene was detected only in one Enterococcus faecium and one S. saprophyticus isolate. An amplicon for acrB efflux pump was detected in all but one isolate. When minimal inhibitory concentrations (MICs) were determined, it was found that the addition of reserpine reduced the MICs by eight fold for most of the biocides and isolates, corroborating the role of efflux pumps in biocide resistance. Erythromycin resistance gene ermB was detected in 90% of Bacillus isolates, and in one Staphylococcus, while ereA was detected only in one Bacillus and one Staphyloccus, and ereB only in one Staphylococcus. The ATP-dependent msrA gene (which confers resistance to macrolides, lincosamides and type B streptogramins) was detected in 60% of Bacillus isolates and in all staphylococci, which in addition carried msrB. The lincosamide and streptogramin A resistance gene lsa was detected in Staphylococcus (40%), Bacillus (27.27%) and Enterococcus (8%) isolates. The aminoglycoside resistance determinant aph (3_)-IIIa was detected in Staphylococcus (40%) and Bacillus (one isolate), aph(2_)-1d in Bacillus (27.27%) and Enterococcus (8%), aph(2_)-Ib in Bacillus (one isolate), and the bifunctional aac(6_)1e-aph(2_)-Ia in Staphylococcus (20%), Enterococcus (8%) and Bacillus (one isolate). Chloramphenicol resistance cat gene was detected in Enterococcus (8%) and Staphylococcus (20%), and blaZ only in Staphylococcus (20%). All other antibiotic or biocide resistance genes investigated were not detected in any isolate. Isolates carrying multiple biocide and antibiotic determinants were frequent among Bacillus (36.36%) and Staphylococcus (50%), but not Enterococcus. These results suggest that biocide and antibiotic determinants may be co-selected. PMID:24361832

Fernández-Fuentes, Miguel Angel; Abriouel, Hikmate; Ortega Morente, Elena; Pérez Pulido, Rubén; Gálvez, Antonio

2014-02-17

9

Gram-positive rhizobacterium Bacillus amyloliquefaciens FZB42 colonizes three types of plants in different patterns.  

PubMed

The colonization of three types of different plants, Zea mays, Arabidopsis thaliana, and Lemna minor, by GFP-labeled Gram-positive rhizobacterium Bacillus amyloliquefaciens FZB42 was studied in gnotobiotic systems using confocal laser scanning microscopy and electron microscopy. It was demonstrated that FZB42 was able to colonize all the plants. On one hand, similar to some Gram-negative rhizobacteria like Pseudomonas, FZB42 favored the areas such as the concavities in root surfaces and the junctions where lateral roots occurred from the primary roots; on the other hand, we clearly demonstrated that root hairs were a popular habitat to the Gram-positive rhizobacterium. FZB42 exhibited a specific colonization pattern on each of the three types of plants. On Arabidopsis, tips of primary roots were favored by FZB42 but not so on maize. On Lemna, FZB42 accumulated preferably along the grooves between epidermal cells of roots and in the concave spaces on ventral sides of fronds. The results suggested L. minor to be a promising tool for investigations on plant-microbial interaction due to a series of advantages it has. Colonization of maize and Arabidopsis roots by FZB42 was also studied in the soil system. Comparatively, higher amount of FZB42 inoculum (?10(8) CFU/ml) was required for detectable root colonization in the soil system, where the preference of FZB42 cells to root hairs were also observed. PMID:22367935

Fan, Ben; Borriss, Rainer; Bleiss, Wilfrid; Wu, Xiaoqin

2012-02-01

10

Extracellular vesicles produced by the Gram-positive bacterium Bacillus subtilis are disrupted by the lipopeptide surfactin.  

PubMed

Previously, extracellular vesicle production in Gram-positive bacteria was dismissed due to the absence of an outer membrane, where Gram-negative vesicles originate, and the difficulty in envisioning how such a process could occur through the cell wall. However, recent work has shown that Gram-positive bacteria produce extracellular vesicles and that the vesicles are biologically active. In this study, we show that Bacillus subtilis produces extracellular vesicles similar in size and morphology to other bacteria, characterized vesicles using a variety of techniques, provide evidence that these vesicles are actively produced by cells, show differences in vesicle production between strains, and identified a mechanism for such differences based on vesicle disruption. We found that in wild strains of B. subtilis, surfactin disrupted vesicles while in laboratory strains harbouring a mutation in the gene sfp, vesicles accumulated in the culture supernatant. Surfactin not only lysed B. subtilis vesicles, but also vesicles from Bacillus anthracis, indicating a mechanism that crossed species boundaries. To our knowledge, this is the first time a gene and a mechanism has been identified in the active disruption of extracellular vesicles and subsequent release of vesicular cargo in Gram-positive bacteria. We also identify a new mechanism of action for surfactin. PMID:24826903

Brown, Lisa; Kessler, Anne; Cabezas-Sanchez, Pablo; Luque-Garcia, Jose L; Casadevall, Arturo

2014-07-01

11

Volatile organic compound analysis by ion molecule reaction mass spectrometry for Gram-positive bacteria differentiation.  

PubMed

Approximately 50 % of all clinically proven infections in critically ill patients are caused by Gram-positive bacteria. The timely and appropriate treatment of these infections is vital in order to avoid negative outcomes. Hence, fast and reliable methods are needed for the early detection and identification of microorganisms. Recently, direct mass spectrometry-based analysis of volatile organic compounds emitted by microorganisms has been employed to study Gram-negative bacteria. Here, we report a feasibility study of ion molecule reaction mass spectrometry (IMR-MS) for in vitro growth detection and species differentiation of selected Gram-positive bacteria that are frequently isolated in blood culture samples, namely, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, and Staphylococcus epidermidis. Ion molecule reaction mass spectrometry was used to analyze the headspace above cultures containing Gram-positive bacteria incubated at 37 °C starting with 10(2) colony-forming units (CFU)/ml. Measurements to determine the presence of volatile organic compounds were performed 4, 8, and 24 h after incubation, respectively. The detection of microbial growth was accomplished already after 8 h in cultures containing E. faecalis. After 24 h of incubation, characteristic mass spectra were obtained for all species. Processing these mass spectra by hierarchic clustering and principal component analysis (PCA) enabled us to differentiate between bacterial species. IMR-MS in conjunction with a cumulative end-point model provides the means for rapid growth detection and differentiation of Gram-positive bacteria on the species level, typically within an analysis time of less than 3 min per sample. PMID:22782437

Dolch, M E; Hornuss, C; Klocke, C; Praun, S; Villinger, J; Denzer, W; Schelling, G; Schubert, S

2012-11-01

12

Proteomics of arsenic stress in the gram-positive organism Exiguobacterium sp. PS NCIM 5463.  

PubMed

The general responses of microorganisms to environmental onslaughts are modulated by altering the gene expression pattern to reduce damage in the cell and produce compensating stress responses. The present study attempts to unravel the response of the Gram-positive Exiguobacterium sp. PS NCIM 5463 in the presence of [As(III)] and arsenate [As(V)] using comparative proteomics via two-dimension gel electrophoresis (2-DE) coupled with identification of proteins using matrix-assisted laser desorption/ionisation (MALDI-TOF/MALDI-TOF/TOF). Out of 926 Coomassie-stained proteins, 45 were differentially expressed (p?organisms, the present study identified new proteins under arsenic stress in a Gram-positive organism, Exiguobacterium sp. PS NCIM 5463, which could elucidate the physiology of organisms under arsenic stress. PMID:24931308

Sacheti, Poonam; Patil, Rajendra; Dube, Ankita; Bhonsle, Hemangi; Thombre, Dipalee; Marathe, Sayali; Vidhate, Ravindra; Wagh, Priyanka; Kulkarni, Mahesh; Rapole, Srikanth; Gade, Wasudev

2014-08-01

13

Performance of the Verigene Gram-Positive Blood Culture Assay for Direct Detection of Gram-Positive Organisms and Resistance Markers in a Pediatric Hospital  

PubMed Central

The performance characteristics of the Verigene Gram-positive blood culture (BC-GP) assay were evaluated in pediatric patients. Concordance of the BC-GP assay was 95.8%, with significant decreases in turnaround time for identification and resistance detection. BC-GP is highly accurate and can be integrated into the routine workflow of the microbiology laboratory.

Mestas, Javier; Polanco, Claudia M.; Felsenstein, Susanna

2014-01-01

14

Modeling of rare earth element sorption to the Gram positive Bacillus subtilis bacteria surface.  

PubMed

In this study, rare earth element (REE) binding constants and site concentration on the Gram+ bacteria surfaces were quantified using a multi-site Langmuir isotherm model, along with a linear programming regression method (LPM), applied to fit experimental REE sorption data. This approach found one discrete REE binding site on the Gram+ Bacillus subtilis surface for the pH range of 2.5-4.5. Average log10 REE binding constants for a site j on these bacteria ranged from 1.08±0.04 to 1.40±0.04 for the light REE (LREE: La to Eu), and from 1.36±0.03 to 2.18±0.14 for the heavy REE (HREE: Gd to Lu) at the highest biomass concentration of 1.3 g/L of B. subtilis bacteria. Similar values were obtained for bacteria concentrations of 0.39 and 0.67 g/L indicating the independence of REE sorption constants on biomass concentration. Within the experimental pH range in this study, B. subtilis was shown to have a lower affinity for LREE (e.g. La, Ce, Pr, Nd) and a higher affinity for HREE (e.g. Tm, Yb, Lu) suggesting an enrichment of HREE on the surface of Gram+ bacteria. Total surface binding site concentrations of 6.73±0.06 to 5.67±0.06 and 5.53±0.07 to 4.54±0.03 mol/g of bacteria were observed for LREE and HREE respectively, with the exception of Y, which showed a total site concentration of 9.53±0.03, and a log K(REE,j) of 1.46±0.02 for a biomass content of 1.3 g/L. The difference in these values (e.g. a lower affinity and increased binding site concentration for LREE, and the contrary for the HREE) suggests a distinction between the LREE and HREE binding modes to the Gram+ bacteria reactive surface at low pH. This further implies that HREE may bind more than one monoprotic reactive group on the cell surface. A multisite Langmuir isotherm approach along with the LPM regression method, not requiring prior knowledge of the number or concentration of cell surface REE complexation sites, were able to distinguish between the sorption constant and binding site concentration patterns of LREE and HREE on the Gram+ B. subtilis surface. This approach quantified the enrichment of Tm, Yb and Lu on the bacteria surface and it has therefore proven to be a useful tool for the study of natural reactive sorbent materials controlling REE partitioning in the natural environment. PMID:24183437

Martinez, Raul E; Pourret, Olivier; Takahashi, Yoshio

2014-01-01

15

A pathway closely related to the (D)-tagatose pathway of gram-negative enterobacteria identified in the gram-positive bacterium Bacillus licheniformis.  

PubMed

We report the first identification of a gene cluster involved in d-tagatose catabolism in Bacillus licheniformis. The pathway is closely related to the d-tagatose pathway of the Gram-negative bacterium Klebsiella oxytoca, in contrast to the d-tagatose 6-phosphate pathway described in the Gram-positive bacterium Staphylococcus aureus. PMID:23524682

Van der Heiden, Edwige; Delmarcelle, Michaël; Lebrun, Sarah; Freichels, Régine; Brans, Alain; Vastenavond, Christian M; Galleni, Moreno; Joris, Bernard

2013-06-01

16

Development of an Organ Culture System for the Study of an Invasive Gram Positive Organism Infecting Bullfrogs, 'Rana catesbeiana'.  

National Technical Information Service (NTIS)

Recent observations on frogs afflicted with red-leg disease indicate that the clinically important bacteria associated with this disease include 13 gram negative bacteria and one gram positive bacterium. Attempts to study the role of the gram positive iso...

R. L. Amborski J. C. Glorioso G. F. Amborski

1974-01-01

17

Structure analysis of a class II transposon encoding the mercury resistance of the Gram-positive bacterium Bacillus megaterium MB1, a strain isolated from Minamata Bay, Japan  

Microsoft Academic Search

A unique transposon was found in the chromosome of Bacillus megaterium MB1, a Gram-positive bacterium isolated from mercury-polluted sediments of Minamata Bay, Japan. The transposon region of a 14.5kb DNA fragment was amplified by PCR using a single PCR primer designed from the nucleotide sequence of an inverted repeat of class II transposons. The molecular analysis revealed that the PCR-amplified

Chieh-Chen Huang; Masaru Narita; Takeshi Yamagata; Yukihiro Itoh; Ginro Endo

1999-01-01

18

Novel Fastidious, Partially Acid-Fast, Anaerobic Gram-Positive Bacillus Associated with Abscess Formation and Recovered from Multiple Medical Centers  

PubMed Central

We report a novel anaerobe causing abscess in four patients at three hospitals. In the clinical specimen, bacilli were branching, Gram positive, and acid fast. The organism grew slowly and was not identified by 16S rRNA sequencing. Our findings support the description of a new genus and species of the suborder Corynebacterineae.

Bell, M.; Bernard, K.; Lagace-Wiens, P.; Schuetz, A. N.; Hartman, B.; McQuiston, J. R.; Wilson, D.; LaSalvia, M.; Ng, B.; Richter, S.; Taege, A.

2013-01-01

19

Characterization of S. pneumoniae pneumonia-induced multiple organ dysfunction syndrome: an experimental mouse model of gram-positive sepsis.  

PubMed

Streptococcus pneumoniae, a gram-positive bacteria, is the most common cause of community-acquired pneumonia. It is a common cause of septic shock with multiple organ dysfunction syndrome (MODS) resulting in significant mortality. Gram-positive mouse models of sepsis with MODS are required to examine mechanisms of immune responses in severe sepsis. To assess whether lung infection due to S. pneumoniae in a nonventilated mouse model can induce multiple organ dysfunction. S. pneumoniae, SPN 15814 strain, harvested at log phase, was injected intratracheally in C57BL/6 mice at OD 600 between 0.35 and 0.63. A dose of bacteria at OD 600 = 0.63 conferred approximately 30% mortality in 36 h. Lung pneumonia was assessed by histology, lung myeloperoxidase activity, and lung bacterial load; intestinal epithelial barrier integrity was assessed by measuring blood-to-lumen clearance of Cr-EDTA; renal function was assessed by measuring plasma creatinine and urea; and myocardiac function was assessed using an isolated perfused mouse heart model. S. pneumoniae-induced pneumonia resulted in neutrophil infiltration into the lungs and increased lung bacterial load. Although relatively few bacteria gained access to the blood stream, the pneumonia was accompanied by increased intestinal epithelial barrier permeability, increased plasma creatinine, and decreased cardiac output and stroke volume. These data clearly show that intratracheal S. pneumoniae induced not only pneumonia but also MODS, despite the fact that few organisms gain access to the blood stream. This model can be used as a good gram-positive model of sepsis and MODS for further studies. PMID:18827750

Andonegui, Graciela; Goring, Kim; Liu, Dan; McCafferty, Donna-Marie; Winston, Brent W

2009-04-01

20

Expanding the use of a fluorogenic method to determine activity and mode of action of Bacillus thuringiensis bacteriocins against Gram-positive and Gram-negative bacteria.  

PubMed

Previously we described a rapid fluorogenic method to measure the activity of five bacteriocins produced by Mexican strains of Bacillus thuringiensis against B. cereus 183. Here we standardize this method to efficiently determine the activity of bacteriocins against both Gram-positive and Gram-negative bacteria. It was determined that the crucial parameter required to obtain reproducible results was the number of cells used in the assay, that is, ~4 × 10(8) cell/mL and ~7 × 10(8) cell/mL, respectively, for target Gram-positive and Gram-negative bacteria. Comparative analyses of the fluorogenic and traditional well-diffusion assays showed correlation coefficients of 0.88 to 0.99 and 0.83 to 0.99, respectively, for Gram-positive and Gram-negative bacteria. The fluorogenic method demonstrated that the five bacteriocins of B. thuringiensis have bacteriolytic and bacteriostatic activities against all microorganisms tested, including clinically significant bacteria such as Listeria monocytogenes, Proteus vulgaris, and Shigella flexneri reported previously to be resistant to the antimicrobials as determined using the well-diffusion protocol. These results demonstrate that the fluorogenic assay is a more sensitive, reliable, and rapid method when compared with the well-diffusion method and can easily be adapted in screening protocols for bacteriocin production by other microorganisms. PMID:22919330

de la Fuente-Salcido, Norma M; Barboza-Corona, J Eleazar; Espino Monzón, A N; Pacheco Cano, R D; Balagurusamy, N; Bideshi, Dennis K; Salcedo-Hernández, Rubén

2012-01-01

21

Expanding the Use of a Fluorogenic Method to Determine Activity and Mode of Action of Bacillus thuringiensis Bacteriocins Against Gram-Positive and Gram-Negative Bacteria  

PubMed Central

Previously we described a rapid fluorogenic method to measure the activity of five bacteriocins produced by Mexican strains of Bacillus thuringiensis against B. cereus 183. Here we standardize this method to efficiently determine the activity of bacteriocins against both Gram-positive and Gram-negative bacteria. It was determined that the crucial parameter required to obtain reproducible results was the number of cells used in the assay, that is, ~4?×?108?cell/mL and ~7?×?108?cell/mL, respectively, for target Gram-positive and Gram-negative bacteria. Comparative analyses of the fluorogenic and traditional well-diffusion assays showed correlation coefficients of 0.88 to 0.99 and 0.83 to 0.99, respectively, for Gram-positive and Gram-negative bacteria. The fluorogenic method demonstrated that the five bacteriocins of B. thuringiensis have bacteriolytic and bacteriostatic activities against all microorganisms tested, including clinically significant bacteria such as Listeria monocytogenes, Proteus vulgaris, and Shigella flexneri reported previously to be resistant to the antimicrobials as determined using the well-diffusion protocol. These results demonstrate that the fluorogenic assay is a more sensitive, reliable, and rapid method when compared with the well-diffusion method and can easily be adapted in screening protocols for bacteriocin production by other microorganisms.

de la Fuente-Salcido, Norma M.; Barboza-Corona, J. Eleazar; Espino Monzon, A. N.; Pacheco Cano, R. D.; Balagurusamy, N.; Bideshi, Dennis K.; Salcedo-Hernandez, Ruben

2012-01-01

22

Regulated RNA stability in the Gram positives  

PubMed Central

Regulation of bacterial gene expression at the post-transcriptional level has emerged as a major control mechanism, although not yet as well recognized as the mechanisms of control at the transcriptional level. In this article, we focus on regulated RNA decay in the control of gene expression in Gram-positive organisms, with a focus on Bacillus subtilis. Discovery of new ribonuclease activities in B. subtilis and other Gram-positive species, especially the dual-functioning RNase J1, which specifies both an endonuclease activity and the long-sought bacterial 5’-to-3’ exoribonuclease activity, has led to the recognition of intriguing mechanisms of gene regulation at the level of RNA decay.

Condon, Ciaran; Bechhofer, David H.

2011-01-01

23

Integrated Physical and Genetic Mapping of Bacillus cereus and Other Gram-Positive Bacteria Based on IS231A Transposition Vectors  

PubMed Central

The genome structure of Bacillus cereus is relatively complex, its DNA being modulated between a size-varying chromosome and large plasmids. To study the genetic organization of the B. cereus type strain ATCC 14579, thermosensitive transposition vectors were designed on the basis of IS231A-derived cassettes containing uncommon restriction sites. A highly preferred insertion site for IS231A was detected in the chromosome by Southern blotting and pulsed-field gel electrophoresis (PFGE) analyses of independent insertion mutants. However, once this insertional hot spot was occupied, secondary IS231A insertions occurred randomly, as demonstrated by isolation of independent B. cereus auxotrophs at a frequency of approximately 0.6%. The hot-spot site, as well as several auxotrophic mutations, were mapped by using NotI, SfiI, and AscI PFGE restriction profiles. It was confirmed by sequencing that one of the insertions, generating an Ade? phenotype, had disrupted a gene of the purine synthesis pathway. These results showed that combined PFGE and sequencing analyses of mini-IS231A insertions enable the construction of integrated physical and genetic maps of B. cereus type strain. Moreover, the presence of the ultrarare I-SceI restriction site in the mini-IS231A allowed the isolation, in double-insertion mutants, of contiguous and nonoverlapping large chromosomal fragments, convenient for direct sequencing. The system detailed in this report is therefore a powerful tool for comparative genetic studies among members of the B. cereus group (i.e., B. cereus, B. thuringiensis, B. mycoides, and B. anthracis) and could also be applied to more distantly related gram-positive bacteria.

Leonard, Catherine; Zekri, Omar; Mahillon, Jacques

1998-01-01

24

Rapid Detection of Gram-Positive Organisms by Use of the Verigene Gram-Positive Blood Culture Nucleic Acid Test and the BacT/Alert Pediatric FAN System in a Multicenter Pediatric Evaluation  

PubMed Central

Assays that expedite the reporting of organism identification and antibiotic susceptibility status in positive blood cultures can fast track interventions that improve clinical outcomes. We evaluated the Verigene Gram-positive blood culture nucleic acid test (BC-GP) in two pediatric hospitals. Positive BacT/Alert Pediatric FAN blood cultures with Gram-positive organisms were tested using the BC-GP in tandem with routine laboratory procedures. To test organisms underrepresented in the clinical blood culture evaluation, blood culture bottles were spiked with diluted organism suspensions at concentrations of 10 to 100 CFU per milliliter. A total of 249 Gram-positive bacterial isolates were recovered from 242 blood cultures. The BC-GP detected Staphylococcus aureus, methicillin-susceptible S. aureus, and methicillin-resistant S. aureus with sensitivities of 100%, 99%, and 100% and specificities of 100%, 100%, and 99.5%, respectively. The BC-GP detected Staphylococcus epidermidis, methicillin-susceptible S. epidermidis, and methicillin-resistant S. epidermidis with sensitivities of 95%, 80%, and 96%, respectively, and 100% specificity. The BC-GP correctly identified 14/15 cases of Enterococcus faecalis and Enterococcus faecium bacteremia and 9 cases of Streptococcus pneumoniae. It misidentified 5/15 clinical blood cultures with Streptococcus mitis/Streptococcus oralis and 1/3 blood cultures spiked with Streptococcus anginosus group as S. pneumoniae. The BC-GP detected a case of Streptococcus pyogenes bacteremia but failed to detect 2/3 clinical blood cultures with Streptococcus agalactiae. BC-GP's rapid accurate detection of Staphylococcus spp., E. faecium, and E. faecalis and its ability to ascertain mecA, vanA, and vanB status may expedite clinical decisions pertaining to optimal antibiotic use. False-positive S. pneumoniae results may warrant reporting of only “Streptococcus spp.” when this organism is reported by the BC-GP.

Turner, N. N.; Roundtree, S. S.; Young, S.; Brock-Haag, C. A.; Lacey, D.; Abuzaid, S.; Blecker-Shelly, D. L.; Doern, C. D.

2013-01-01

25

Rapid detection of Gram-positive organisms by use of the Verigene Gram-positive blood culture nucleic acid test and the BacT/Alert Pediatric FAN system in a multicenter pediatric evaluation.  

PubMed

Assays that expedite the reporting of organism identification and antibiotic susceptibility status in positive blood cultures can fast track interventions that improve clinical outcomes. We evaluated the Verigene Gram-positive blood culture nucleic acid test (BC-GP) in two pediatric hospitals. Positive BacT/Alert Pediatric FAN blood cultures with Gram-positive organisms were tested using the BC-GP in tandem with routine laboratory procedures. To test organisms underrepresented in the clinical blood culture evaluation, blood culture bottles were spiked with diluted organism suspensions at concentrations of 10 to 100 CFU per milliliter. A total of 249 Gram-positive bacterial isolates were recovered from 242 blood cultures. The BC-GP detected Staphylococcus aureus, methicillin-susceptible S. aureus, and methicillin-resistant S. aureus with sensitivities of 100%, 99%, and 100% and specificities of 100%, 100%, and 99.5%, respectively. The BC-GP detected Staphylococcus epidermidis, methicillin-susceptible S. epidermidis, and methicillin-resistant S. epidermidis with sensitivities of 95%, 80%, and 96%, respectively, and 100% specificity. The BC-GP correctly identified 14/15 cases of Enterococcus faecalis and Enterococcus faecium bacteremia and 9 cases of Streptococcus pneumoniae. It misidentified 5/15 clinical blood cultures with Streptococcus mitis/Streptococcus oralis and 1/3 blood cultures spiked with Streptococcus anginosus group as S. pneumoniae. The BC-GP detected a case of Streptococcus pyogenes bacteremia but failed to detect 2/3 clinical blood cultures with Streptococcus agalactiae. BC-GP's rapid accurate detection of Staphylococcus spp., E. faecium, and E. faecalis and its ability to ascertain mecA, vanA, and vanB status may expedite clinical decisions pertaining to optimal antibiotic use. False-positive S. pneumoniae results may warrant reporting of only "Streptococcus spp." when this organism is reported by the BC-GP. PMID:23966484

Sullivan, K V; Turner, N N; Roundtree, S S; Young, S; Brock-Haag, C A; Lacey, D; Abuzaid, S; Blecker-Shelly, D L; Doern, C D

2013-11-01

26

Purification and characterization of two distinct thermostable lipases from the gram-positive thermophilic bacterium Bacillus thermoleovorans ID1  

Microsoft Academic Search

The thermophilic bacterium Bacillus thermoleovorans ID-1 can hydrolyze a variety of oils such as olive oil, soybean oil, palm oil, and lard as a carbon source (1.5%, v\\/v) after 72 h of culture at 50°C. In this study, we purified to homogeneity two distinct thermostable lipases, designated BTID-A (B. thermoleovorans ID-1 lipase A) and BTID-B (B. thermoleovorans ID-1 lipase B).

Dong-Woo Lee; Hack-Woo Kim; Keun-Wook Lee; Byoung-Chan Kim; Eun-Ah Choe; Han-Seung Lee; Doo-Sik Kim; Yu-Ryang Pyun

2001-01-01

27

The influence of secretory-protein charge on late stages of secretion from the Gram-positive bacterium Bacillus subtilis.  

PubMed Central

Following their secretion across the cytoplasmic membrane, processed secretory proteins of Bacillus subtilis must fold into their native conformation prior to translocation through the cell wall and release into the culture medium. The rate and efficiency of folding are critical in determining the yields of intact secretory proteins. The B. subtilis membrane is surrounded by a thick cell wall comprising a heteropolymeric matrix of peptidoglycan and anionic polymers. The latter confer a high density of negative charge on the wall, endowing it with ion-exchange properties, and secretory proteins destined for the culture medium must traverse the wall as the last stage in the export process. To determine the influence of charge on late stages in the secretion of proteins from this bacterium, we have used sequence data from two related alpha-amylases, to engineer the net charge of AmyL, an alpha-amylase from Bacillus licheniformis that is normally secreted efficiently from B. subtilis. While AmyL has a pI of 7.0, chimaeric enzymes with pI values of 5.0 and 10.0 were produced and characterized. Despite the engineered changes to their physico-chemical properties, the chimaeric enzymes retained many of the enzymic characteristics of AmyL. We show that the positively charged protein interacts with the cell wall in a manner that influences its secretion.

Stephenson, K; Jensen, C L; J?rgensen, S T; Lakey, J H; Harwood, C R

2000-01-01

28

Susceptibility of Fibronectin to Degradation by Various Gram-Negative and Gram-Positive Oral Micro-Organisms.  

National Technical Information Service (NTIS)

The degradative effects on tritiated human plasma fibronectin (FN) of proteases associated with twenty-five Gram-negative (GN) and fifteen Gram-positive (GP) oral microbial isolates were examined. Ninety-two percent of the GN and 20% of the GP isolates de...

E. D. Pederson B. L. Lamberts I. L. Shklair

1988-01-01

29

Evaluation of a Microarray-Based Assay for Rapid Identification of Gram-Positive Organisms and Resistance Markers in Positive Blood Cultures  

PubMed Central

Rapid identification of pathogens directly from positive blood cultures can play a major role in reducing patient mortality rates. We evaluated the performance of the Verigene Gram-Positive Blood Culture (BC-GP) assay (Nanosphere Inc., Northbrook, IL) for detection of commonly isolated Gram-positive organisms as well as associated resistance markers from positive blood cultures. Positive blood cultures (VersaTREK; Trek Diagnostic Systems, Independence, OH) from 203 patients with Gram-positive organism infections were analyzed using the BC-GP assay within 12 h for the detection of 12 different organisms, including staphylococci, streptococci, and enterococci, as well as for the presence of 3 resistance markers (mecA, vanA, and vanB). Results were compared to those of routine laboratory methods for identification and susceptibility testing. For identification of organisms and detection of resistance markers in 178 monomicrobial positive blood cultures, the BC-GP assay showed 94% and 97% concordance, respectively, with routine methods. After 25 polymicrobial cultures were included, the results showed 92% and 96% agreement for identification and resistance markers, respectively, for a total of 203 positive cultures. In 6/25 polymicrobial cultures, at least 1 isolate was not detected. Concordance levels for detection of major pathogens such Staphylococcus aureus (n = 45) and enterococci (n = 19) were 98% and 95%, respectively. Agreement levels for detection of resistance markers such as mecA and vanA/B were 92% and 100%, respectively. The BC-GP assay is capable of providing rapid identification of Gram-positive cocci as well as detection of resistance markers directly from positive blood cultures at least 24 to 48 h earlier than conventional methods.

Tibbetts, Robert J.; Agotesku, Adam; Fey, Margaret; Hensley, Rhonda; Meier, Frederick A.

2013-01-01

30

Isolation of a Gram-positive poly(3-hydroxybutyrate) (PHB)-degrading bacterium from compost, and cloning and characterization of a gene encoding PHB depolymerase of Bacillus megaterium N-18-25-9.  

PubMed

A Gram-positive poly(3-hydroxybutyrate) (PHB)-degrading bacterial strain was isolated from compost. This organism, identified as Bacillus megaterium N-18-25-9, produced a clearing zone on opaque NB-PHB agar, indicating the presence of extracellular PHB depolymerase. A PHB depolymerase gene, PhaZ(Bm), of B. megaterium N-18-25-9 was cloned and sequenced, and the recombinant gene product was purified from Escherichia coli. The N-terminal half region of PhaZ(Bm) shared significant homologies with a catalytic domain of other PHB depolymerases. Although the C-terminal half region of PhaZ(Bm) showed no significant similarity with those of other PHB depolymerases, that region was necessary for the PHB depolymerase activity. Therefore, this enzyme's domain structure is unique among extracellular PHB depolymerase domain structures. The addition of PHB to the medium led to a sixfold increase in PhaZ(Bm) mRNA, while the presence of glucose repressed PhaZ(Bm) expression. The maximum activity was observed at pH 9.0 at 65 degrees C. PMID:17064368

Takaku, Hiroaki; Kimoto, Ayumi; Kodaira, Shoko; Nashimoto, Masayuki; Takagi, Masamichi

2006-11-01

31

Isolation, characterization, and biological properties of an endotoxin-like material from the gram-positive organism Listeria monocytogenes.  

PubMed Central

The bacterial component responsible for the induction of transient cold agglutinin syndrome in rabbits after intravenous injection of heat-killed Listeria monocytogenes type 4B has been purified and biologically and chemically characterized. A purified immunoglobulin M cold agglutinin was prepared from high-titer sera resulting from the immunization of rabbits with heat-killed L. monocytogenes type 4B and was subsequently used to monitor the purification of the bacterial component responsible for its induction. The bacterial component was isolated from a hot phenol-water extract of lyophilized L. monocytogenes type 4B by multiple molecular sieve chromatography. Upon chemical analysis the purified material was found to be strikingly similar in chemical composition to gram-negative lipopolysaccharide endotoxins. The material contained 15% total fatty acid (of which 50% was beta-hydroxymyristic acid), 40 to 45% neutral sugar (glucose, galactose, and rhamnose), 11.5% amino sugar, 12% uronic acid, 2.5% 2-keto-3-deoxyoctonic acid, 2% heptose, 0.87% phosphorus, and 1.6% amino acid, thereby accounting for 85 to 90% of the weight of the component. Electron micrographs of the purified material were similar to those of lipopolysaccharide preparations from gram-negative organisms. The purified material exist in aqueous solutions as large aggregates, but can be dissociated into a single smaller subunit (3.1S) by dialysis against sodium dodecyl sulfate buffer. The listerial component was toxic and pyrogenic to rabbits, producing symptoms typical of gram-negative endotoxins. Activity in the limulus lysate gelation assay and in the carbocyanine dye assay provides a further link of this material with classical gram-negative endotoxins. Images

Wexler, H; Oppenheim, J D

1979-01-01

32

Isolation of insertion elements from Gram-positive Brevibacterium, Corynebacterium and Rhodococcus strains using the Bacillus subtilis sacB gene as a positive selection marker  

Microsoft Academic Search

The sacB gene of Bacillus subtilis was successfully applied in various Arthrobacter, Brevibacterium, Corynebacterium and Rhodococcus strains for the isolation of transposable elements. Three different insertion sequence (IS) elements entrapped in sacB were isolated. The IS elements IS-Bl and IS-Cg isolated from Brevibacterium lactofermentum and Corynebacterium glutamicum, respectively, were found to be similar in size (1.45 kb) and generated target

Wolfgang Jäger; Andreas Schäfer; Jörn Kalinowski; Alfred Fühler

1995-01-01

33

Immunological crossreactivity to the catabolite control protein CcpA from Bacillus megaterium is found in many Gram-positive bacteria  

Microsoft Academic Search

The catabolite control protein CcpA from Bacillus megaterium was overproduced as a fusion protein to a 6xhis affinity tag and purified to homogeneity. Polyclonal antibodies of high affinity and specificity were raised against the purified protein. The serum did not crossreact with purified Lac repressor despite the fact that CcpA and LacI belong to the same protein family. Using this

Elke Küster; Evert J. Luesink; Willem M. de Vos; Wolfgang Hillen

1996-01-01

34

Isolation of insertion elements from gram-positive Brevibacterium, Corynebacterium and Rhodococcus strains using the Bacillus subtilis sacB gene as a positive selection marker.  

PubMed

The sacB gene of Bacillus subtilis was successfully applied in various Arthrobacter, Brevibacterium, Corynebacterium and Rhodococcus strains for the isolation of transposable elements. Three different insertion sequence (IS) elements entrapped in sacB were isolated. The IS elements IS-Bl and IS-Cg isolated from Brevibacterium lactofermentum and Corynebacterium glutamicum, respectively, were found to be similar in size (1.45 kb) and generated target duplications of 8 bp. Their inverted repeats showed homology. In contrast, the IS element IS-Rf isolated from Rhodococcus fascians was only 1.3 kb long and generated a 3-bp target duplication. IS-Cg and IS-Rf were not restricted to their original host strains, and we also found strains harbouring more than one element. PMID:7896070

Jäger, W; Schäfer, A; Kalinowski, J; Pühler, A

1995-02-01

35

The replication terminator protein of the gram-positive bacterium Bacillus subtilis functions as a polar contrahelicase in gram-negative Escherichia coli.  

PubMed

The replication terminator protein (RTP) of Bacillus subtilis is a dimer with a monomeric molecular mass of 14.5 kDa. The protein terminates DNA replication at a specific binding site. Although the protein has been crystallized and its crystal structure has been solved, the lack of an in vitro replication system in B. subtilis has been a serious impediment to the analysis of the mechanism of action of this protein. We have discovered that the protein is functional in the Gram-negative bacterium Escherichia coli in vivo and in vitro. RTP blocked replication forks initiated from a ColE1 replication origin at the cognate DNA-binding site (BS3) in a polar mode. The protein did not block rolling circle replication initiated from the pT181 origin in cell extracts of Staphylococcus aureus. RTP antagonized the helicase activity of DnaB but not that of helicase II of E. coli. Thus, RTP functioned as a polar contrahelicase blocking a helicase that participates in symmetric DNA replication but it did not impede rolling circle replication nor the action of a helicase involved in DNA repair. PMID:7972025

Kaul, S; Mohanty, B K; Sahoo, T; Patel, I; Khan, S A; Bastia, D

1994-11-01

36

Pharmacodynamic Properties of BAY 12-8039 on Gram-Positive and Gram-Negative Organisms as Demonstrated by Studies of Time-Kill Kinetics and Postantibiotic Effect  

Microsoft Academic Search

BAY 12-8039 is a novel fluoroquinolone with activity against gram-positive and gram-negative bacteria but not Pseudomo- nas aeruginosa (4). It has been suggested that the bactericidal activities of fluoroquinolones require the presence of oxygen (9), although reports of the activities of quinolones under an- aerobic conditions are conflicting. The pharmacodynamics of an antibiotic may be investigated in several ways, including

F. J. BOSWELL; J. M. ANDREWS; R. WISE

1997-01-01

37

Antimicrobial Susceptibility among Gram-Positive Organisms Collected from Pediatric Patients Globally between 2004 and 2011: Results from the Tigecycline Evaluation and Surveillance Trial  

PubMed Central

The Tigecycline Evaluation and Surveillance Trial (TEST) was designed to monitor global longitudinal changes in bacterial susceptibility to a panel of antimicrobial agents, including tigecycline. In this study, we examine susceptibility among Gram-positive isolates collected from pediatric patients globally between 2004 and 2011. A total of 9,422 Gram-positive isolates were contributed by 1,255 centers, predominantly from Europe and North America. One-third of Staphylococcus aureus isolates were methicillin resistant, peaking in prevalence in 2007. All S. aureus isolates (n = 3,614) were susceptible to linezolid, tigecycline, and vancomycin; minocycline, imipenem, and meropenem were also highly active (>92% susceptibility). Ampicillin and penicillin susceptibility increased significantly during the study period (P < 0.0001 for both). Streptococcus pneumoniae isolates (n = 3,373) were highly susceptible to vancomycin (100%), linezolid (>99%), and levofloxacin and tigecycline (both >96%); imipenem susceptibility was low (32%) in Africa while minocycline susceptibility was low in Asia-Pacific Rim (38%). Penicillin resistance occurred in one-fifth of all S. pneumoniae isolates, with penicillin susceptibility ranging from 14% in Africa to 65% in Europe. Streptococcus agalactiae isolates (n = 1,056) were highly susceptible to most antimicrobials, although only 16% were susceptible to minocycline. Enterococcus faecalis isolates (n = 1,112) were highly susceptible (>97%) to ampicillin, linezolid, penicillin, tigecycline, and vancomycin globally, but only 34% were minocycline susceptible; minocycline susceptibility decreased significantly from 2004 to 2011 (P < 0.001). Tigecycline and linezolid were highly active against Enterococcus faecium (n = 267) globally (100% and 98% susceptible, respectively). Tigecycline and linezolid were highly active against Gram-positive pathogens from pediatric patients in TEST 2004 to 2011, with vancomycin and the carbapenems performing well against most pathogens.

Dowzicky, Michael J.

2013-01-01

38

Streptococcus mutans: a new Gram-positive paradigm?  

PubMed Central

Despite the enormous contributions of the bacterial paradigms Escherichia coli and Bacillus subtilis to basic and applied research, it is well known that no single organism can be a perfect representative of all other species. However, given that some bacteria are difficult, or virtually impossible, to cultivate in the laboratory, that some are recalcitrant to genetic and molecular manipulation, and that others can be extremely dangerous to manipulate, the use of model organisms will continue to play an important role in the development of basic research. In particular, model organisms are very useful for providing a better understanding of the biology of closely related species. Here, we discuss how the lifestyle, the availability of suitable in vitro and in vivo systems, and a thorough understanding of the genetics, biochemistry and physiology of the dental pathogen Streptococcus mutans have greatly advanced our understanding of important areas in the field of bacteriology such as interspecies biofilms, competence development and stress responses. In this article, we provide an argument that places S. mutans, an organism that evolved in close association with the human host, as a novel Gram-positive model organism.

Quivey, Robert G.; Koo, Hyun; Abranches, Jacqueline

2013-01-01

39

Moderately halophilic gram-positive bacterial diversity in hypersaline environments.  

PubMed

Moderately halophilic bacteria are microorganisms that grow optimally in media containing 3%-15% (w/v) salt. They are represented by a heterogeneous group of microorganisms included in many different genera. Gram-negative moderately halophilic bacteria have been studied in more detail, but studies on gram-positive species are more scarce. Recent studies carried out by our research group on gram-positive moderate halophiles have permitted clarifying their taxonomic and phylogenetic position and describing new species. Thus, we have isolated six strains from ponds of salterns that show phenotypic and genotypic characteristics similar to those of Nesterenkonia halobia (formerly Micrococcus halobius), a moderately halophilic gram-positive coccus that was described on the basis of a single strain. Our data demonstrate quite clearly that they are members of this species and contribute to a better description of these moderately halophilic cocci. Similarly, a study of a large number of gram-positive moderately halophilic rods that were able to produce endospores led us to describe a new species, designated Bacillus salexigens. Further, isolates grouped in other three phenons, obtained by numerical taxonomy analysis and showing phenotypic features quite similar to those of this species, represent different genomovars, with very low DNA-DNA homology. Although they might represent additional new species, it will be necessary to determine new phenotypic features to differentiate them from previously described Bacillus species. We have also studied the viability of some old enrichments provided by B.E. Volcani, which were set up in 1936. We isolated 31 gram-positive motile endospore-forming rods that, according to their phenotypic characteristics, could represent a new species of the genus Bacillus. PMID:9783177

Ventosa, A; Márquez, M C; Garabito, M J; Arahal, D R

1998-08-01

40

Regulation of nitrogen metabolism in gram-positive bacteria  

Microsoft Academic Search

A search for new members of the TnrA and GlnR regulons, responsible for assimilation of nitrogen in Gram-positive bacteria,\\u000a was performed. Common regulatory signals with consensus sequences ATGTNAWWWWWWWTNACAT and TGTNAWWWWWWWTNACA were identified\\u000a for GlnR and TnrA, respectively. The structure was described and new potential members were found in Bacillus subtilis, B. licheniformis, Geobacillus kaustophilus, and Oceanobacillus iheyensis for the TnrA\\/GlnR

N. A. Doroshchuk; M. S. Gelfand; D. A. Rodionov

2006-01-01

41

Fate study of water-borne gram positive vegetative bacterial cells with Raman microscopy  

Microsoft Academic Search

We present an initial bacterial fate study of Gram positive vegetative cells suspended in water and stored at ambient room temperature via Raman spectroscopy monitoring. Two types of cells were considered for this study: vegetative cells of Bacillus cereus, Bacillus thuringiensis which contain the polyhydroxybutyric acid (PHBA) as an energy storage compound and Bacillus subtlilis cells which do not. The

Jason Guicheteau; Ashish Tripathi; Jennifer Minter; Phillip Wilcox; Steven Christesen

2010-01-01

42

Pathogenicity of anaerobic gram-positive cocci.  

PubMed Central

The pathogenicity of 20 strains of facultative or anaerobic gram-positive cocci (AGPC) was investigated by injecting them alone or mixed with other flora into mice, utilizing the subcutaneous abscess model. Abscesses induced by a mixture of two organisms were uniformly larger than those induced by single organisms. The relationships among seven AGPC strains, eight aerobes, and two Bacteroides spp. were determined by treating the infected animals with antibiotics and observing the effect of therapy directed against one or both organisms present in the abscess. A total of 70 different combinations were tested. As judged by their responses to antimicrobial therapy, facultative cocci or AGPC were relatively more important than the other species in 6 combinations, equally important in 35 combinations, and less important in 29 combinations. The AGPC most often found to be equal to or more important than the other bacteria were Peptococcus magnus, Streptococcus constellatus, and Peptostreptococcus anaerobius. Proteus mirabilis, Escherichia coli, and Staphylococcus aureus were frequently found to be of more importance than the AGPC.

Brook, I; Walker, R I

1984-01-01

43

Architecture and assembly of the Gram-positive cell wall  

PubMed Central

The bacterial cell wall is a mesh polymer of peptidoglycan – linear glycan strands crosslinked by flexible peptides – that determines cell shape and provides physical protection. While the glycan strands in thin “Gram-negative” peptidoglycan are known to run circumferentially around the cell, the architecture of the thicker “Gram-positive” form remains unclear. Using electron cryotomography, here we show that Bacillus subtilis peptidoglycan is a uniformly dense layer with a textured surface. We further show it rips circumferentially, curls and thickens at free edges, and extends longitudinally when denatured. Molecular dynamics simulations show that only atomic models based on the circumferential topology recapitulate the observed curling and thickening, in support of an “inside-to-outside” assembly process. We conclude that instead of being perpendicular to the cell surface or wrapped in coiled cables (two alternative models), the glycan strands in Gram-positive cell walls run circumferentially around the cell just as they do in Gram-negative cells. Together with providing insights into the architecture of the ultimate determinant of cell shape, this study is important because Gram-positive peptidoglycan is an important antibiotic target crucial to the viability of several important rod-shaped pathogens including Bacillus anthracis, Listeria monocytogenes, and Clostridium difficile.

Beeby, Morgan; Gumbart, James C.; Roux, Benoit; Jensen, Grant J.

2013-01-01

44

Antimicrobial Resistance in Gram-Positive Bacteria  

Microsoft Academic Search

Gram-positive bacteria are common causes of bloodstream and other infections in hospitalized patients in the United States, and the percentage of nosocomial bloodstream infections caused by antibiotic-resistant gram-positive bacteria is increasing. Methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) are of particular concern. In the United States, approximately 60% of staphylococcal infections in the intensive care unit are now caused

Louis B. Rice

2006-01-01

45

Widespread Abundance of Functional Bacterial Amyloid in Mycolata and Other Gram-Positive Bacteria?  

PubMed Central

Until recently, extracellular functional bacterial amyloid (FuBA) has been detected and characterized in only a few bacterial species, including Escherichia coli, Salmonella, and the gram-positive organism Streptomyces coelicolor. Here we probed gram-positive bacteria with conformationally specific antibodies and revealed the existence of FuBA in 12 of 14 examined mycolata species, as well as six other distantly related species examined belonging to the phyla Actinobacteria and Firmicutes. Most of the bacteria produced extracellular fimbriae, sometimes copious amounts of them, and in two cases large extracellular fibrils were also produced. In three cases, FuBA was revealed only after extensive removal of extracellular material by saponification, indicating that there is integrated attachment within the cellular envelope. Spores of species in the genera Streptomyces, Bacillus, and Nocardia were all coated with amyloids. FuBA was purified from Gordonia amarae (from the cell envelope) and Geodermatophilus obscurus, and they had the morphology, tinctorial properties, and ?-rich structure typical of amyloid. The presence of approximately 9-nm-wide amyloids in the cell envelope of G. amarae was visualized by transmission electron microscopy analysis. We conclude that amyloid is widespread among gram-positive bacteria and may in many species constitute a hitherto overlooked integral part of the spore and the cellular envelope.

Jordal, Peter Bruun; Dueholm, Morten Simonsen; Larsen, Poul; Petersen, Steen Vang; Enghild, Jan Johannes; Christiansen, Gunna; H?jrup, Peter; Nielsen, Per Halkjaer; Otzen, Daniel Erik

2009-01-01

46

[Regulation of nitrogen metabolism in gram-positive bacteria].  

PubMed

We searched for new members of the TnrA and GlnR regulons controlling assimilation of nitrogen in gram-positive bacteria. We identified the regulatory signals for these transcription factors with consensuses ATGTNAWWWWWWWTNACAT for GlnR and TGTNAWWWWWWWTNACA for TnrA. We described the structure and found new potential members for the TnrA/GlnR regulons in Bacillus subtilis, B. licheniformis, Geobacillus kaustophilus, Oceanobacillus iheyensis, for the TnrA regulon in B. halodurans and for the GlnR regulons in Lactococcus lactis, Lactobacillus plantarum, Streptococcus pyogenes, S. pneumoniae, S. mutans, S. agalactiae, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus and St. epidermidis. PMID:17086994

Doroshchuk, N A; Gel'fand, M S; Rodionov, D A

2006-01-01

47

Ethanol production in Gram-positive microbes  

DOEpatents

The subject invention concerns the transformation of Gram-positive bacteria with heterologous genes which confer upon these microbes the ability to produce ethanol as a fermentation product. Specifically exemplified is the transformation of bacteria with genes, obtainable from Zymomonas mobilis, which encode pyruvate decarboxylase and alcohol dehydrogenase. 2 figs.

Ingram, L.O.; Barbosa-Alleyne, M.D.F.

1999-06-29

48

Mechanical Consequences of Cell-Wall Turnover in the Elongation of a Gram-Positive Bacterium  

PubMed Central

A common feature of walled organisms is their exposure to osmotic forces that challenge the mechanical integrity of cells while driving elongation. Most bacteria rely on their cell wall to bear osmotic stress and determine cell shape. Wall thickness can vary greatly among species, with Gram-positive bacteria having a thicker wall than Gram-negative bacteria. How wall dimensions and mechanical properties are regulated and how they affect growth have not yet been elucidated. To investigate the regulation of wall thickness in the rod-shaped Gram-positive bacterium Bacillus subtilis, we analyzed exponentially growing cells in different media. Using transmission electron and epifluorescence microscopy, we found that wall thickness and strain were maintained even between media that yielded a threefold change in growth rate. To probe mechanisms of elongation, we developed a biophysical model of the Gram-positive wall that balances the mechanical effects of synthesis of new material and removal of old material through hydrolysis. Our results suggest that cells can vary their growth rate without changing wall thickness or strain by maintaining a constant ratio of synthesis and hydrolysis rates. Our model also indicates that steady growth requires wall turnover on the same timescale as elongation, which can be driven primarily by hydrolysis rather than insertion. This perspective of turnover-driven elongation provides mechanistic insight into previous experiments involving mutants whose growth rate was accelerated by the addition of lysozyme or autolysin. Our approach provides a general framework for deconstructing shape maintenance in cells with thick walls by integrating wall mechanics with the kinetics and regulation of synthesis and turnover.

Misra, Gaurav; Rojas, Enrique R.; Gopinathan, Ajay; Huang, Kerwyn Casey

2013-01-01

49

Bacteriocins of gram-positive bacteria.  

PubMed Central

In recent years, a group of antibacterial proteins produced by gram-positive bacteria have attracted great interest in their potential use as food preservatives and as antibacterial agents to combat certain infections due to gram-positive pathogenic bacteria. They are ribosomally synthesized peptides of 30 to less than 60 amino acids, with a narrow to wide antibacterial spectrum against gram-positive bacteria; the antibacterial property is heat stable, and a producer strain displays a degree of specific self-protection against its own antibacterial peptide. In many respects, these proteins are quite different from the colicins and other bacteriocins produced by gram-negative bacteria, yet customarily they also are grouped as bacteriocins. Although a large number of these bacteriocins (or bacteriocin-like inhibitory substances) have been reported, only a few have been studied in detail for their mode of action, amino acid sequence, genetic characteristics, and biosynthesis mechanisms. Nevertheless, in general, they appear to be translated as inactive prepeptides containing an N-terminal leader sequence and a C-terminal propeptide component. During posttranslational modifications, the leader peptide is removed. In addition, depending on the particular type, some amino acids in the propeptide components may undergo either dehydration and thioether ring formation to produce lanthionine and beta-methyl lanthionine (as in lantibiotics) or thio ester ring formation to form cystine (as in thiolbiotics). Some of these steps, as well as the translocation of the molecules through the cytoplasmic membrane and producer self-protection against the homologous bacteriocin, are mediated through specific proteins (enzymes). Limited genetic studies have shown that the structural gene for such a bacteriocin and the genes encoding proteins associated with immunity, translocation, and processing are present in a cluster in either a plasmid, the chromosome, or a transposon. Following posttranslational modification and depending on the pH, the molecules may either be released into the environment or remain bound to the cell wall. The antibacterial action against a sensitive cell of a gram-positive strain is produced principally by destabilization of membrane functions. Under certain conditions, gram-negative bacterial cells can also be sensitive to some of these molecules. By application of site-specific mutagenesis, bacteriocin variants which may differ in their antimicrobial spectrum and physicochemical characteristics can be produced. Research activity in this field has grown remarkably but sometimes with an undisciplined regard for conformity in the definition, naming, and categorization of these molecules and their genetic effectors. Some suggestions for improved standardization of nomenclature are offered.

Jack, R W; Tagg, J R; Ray, B

1995-01-01

50

New thermosensitive plasmid for gram-positive bacteria.  

PubMed Central

We isolated a replication-thermosensitive mutant of the broad-host-range replicon pWV01. The mutant pVE6002 is fully thermosensitive above 35 degrees C in both gram-negative and gram-positive bacteria. Four clustered mutations were identified in the gene encoding the replication protein of pVE6002. The thermosensitive derivative of the related plasmid pE194 carries a mutation in the analogous region but not in the same position. Derivatives of the thermosensitive plasmid convenient for cloning purposes have been constructed. The low shut-off temperature of pVE6002 makes it a useful suicide vector for bacteria which are limited in their own temperature growth range. Using pVE6002 as the delivery vector for a transposon Tn10 derivative in Bacillus subtilis, we observed transposition frequencies of about 1%. Images

Maguin, E; Duwat, P; Hege, T; Ehrlich, D; Gruss, A

1992-01-01

51

REPORT ANTIBACTERIAL ACTIVITY OF OREGANO (ORIGANUM VULGARE LINN.) AGAINST GRAM POSITIVE BACTERIA  

Microsoft Academic Search

The present investigation is focused on antibacterial potential of infusion, decoction and essential oil of oregano (Origanum vulgare) against 111 Gram-positive bacterial isolates belonging to 23 different species related to 3 genera. Infusion and essential oil exhibited antibacterial activity against Staphylococcus saprophyticus, S. aureus, Micrococcus roseus, M. kristinae, M. nishinomiyaensis, M. lylae, M. luteus, M. sedentarius, M. varians, Bacillus megaterium,

SABAHAT SAEED; PERWEEN TARIQ

52

A rapid procedure for isolating chromosomal DNA from Lactobacillus species and other Gram-positive bacteria  

Microsoft Academic Search

R. L. U L R I C H A N D T. A. H U G H E S. 2001. This study describes a rapid procedure for the isolation of genomic DNA from various Gram-positive bacteria. Species tested included Lactobacillus delbrueckii subsp. lactis ATCC 4797, Lact. acidophilus N2, Staphylococcus aureus, Staph. epidermidis, Propionibacterium jensenii P126, Bacillus pumilus and Enterococcus faecalis.

R. L. Ulrich; T. A. Hughes

2001-01-01

53

Nisin inhibits several gram-positive, mastitis-causing pathogens.  

PubMed

Organisms known to cause bovine mastitis, Enterococcus faecalis ssp. liquefaciens ATCC 27959, Staphylococcus aureus ATCC 29740, Streptococcus agalactiae ATCC 27956, Streptococcus equinus ATCC 27960, Streptococcus dysgalactiae ATCC 27957, Streptococcus uberis ATCC 27958, and the neotype Staphylococcus epidermidis ATCC 14990 were examined for their susceptibility to the small peptide antibiotic, nisin. Using a disc assay, minimum inhibitory concentrations of nisin ranged from 10 to 250 micrograms/ml among the strains. Examination of the antimicrobial effect of 50 micrograms/ml nisin in milk showed nisin inhibited all gram-positive pathogens tested. PMID:2516858

Broadbent, J R; Chou, Y C; Gillies, K; Kondo, J K

1989-12-01

54

Salusin-?, an Antimicrobially Active Peptide against Gram-Positive Bacteria.  

PubMed

Salusin-? has been detected in numerous mammalian tissues and has been shown to have various effects on the cardiovascular system. In this study, we showed that salusin-? exhibited potent antibacterial activity against Gram-positive microorganisms such as Bacillus subtilis NBRC 3513, Bacillus megaterium ATCC 19213, Staphylococcus aureus NBRC 12732, and Staphylococcus epidermidis NBRC 12933. A cytoplasmic membrane-depolarizing assay using the DiSC3(5) dye revealed that the addition of 4?nmol/mL of salusin-? caused the leakage of fluorescence dye from Staphylococcus aureus NBRC 12732. The antimicrobial potency and circular dichroism (CD) spectroscopy of five analogs related to salusin-? were examined to determine structure-function relationships in its N- and C-terminal regions. The results obtained suggest that the N-terminal sequences of the salusin-? molecule are important for the expression of the potent antimicrobial activity of this peptide. A profile corresponding to that of the ?-helix conformation was observed in the salusin-? solution. PMID:24881665

Kimura, Masahiro; Shindo, Mitsuno; Moriizumi, Toshiyuki; Tagawa, Noriko; Fujinami, Aya; Kato, Ikuo; Uchida, Yoshiki

2014-01-01

55

A Mysterious Gram-Positive Rods  

PubMed Central

We encountered a patient with a history of intravenous drug use presenting with fever, malaise and nausea who was found to have cavitary lung lesions. Unexpectedly, gram positive rods grew out on day five on multiple blood cultures, which were later identified as Mycobacterium fortuitum. The patient underwent transesophageal echocardiogram, which showed aortic and tricuspid valve vegetations. Liver biopsy demonstrated granulomatous hepatitis. Interestingly, serum alkaline phosphatase level fell with antibiotic treatment. Mycobacterium fortuitum is ubiquitous worldwide, being found in tap water, and soil. M. fortuitum is usually considered as a contaminant. Disseminated infection caused by this bacterium in an immunocompetent host is extremely rare. Most of the disseminated infections have been reported in immune-deficient patients. In immunocompetent people, M. fortuitum causes human infection primarily by direct inoculation, including localized post-traumatic and surgical wound infections, and catheter-related sepsis. Our patient, an HIV-negative intravenous drug user, had Mycobacterium fortuitum sepsis associated with infective endocarditis, septic pulmonary emboli, and granulomatous hepatitis. Interestingly, the patient admitted using tap water occasionally for mixing heroin when her sterile water ran out, which we thought was the likely source of M. fortuitum.

Natsag, Javzandulam; Min, Zaw; Hamad, Yasir; Alkhalil, Bassel; Rahman, Atiq; Williams, Richard

2012-01-01

56

Evaluation of the VITEK 2 gram positive (GP) microbial identification test card: collaborative study.  

PubMed

A collaborative study was conducted to evaluate the performance of the VITEK 2 Gram Positive (GP) identification card for use with the VITEK 2 automated microbial identification system. The GP test card is used in the identification of selected Gram positive organisms, including Listeria and Staphylococcus species. The VITEK 2 GP card is based on 43 biochemical tests measuring carbon source utilization, inhibition and resistance, and enzymatic activities. A total of 20 laboratories representing government, industry, and private testing laboratories throughout the United States participated. In this study, 720 Gram-positive inclusivity isolates were analyzed by the GP Identification method. Of the 720 well-characterized isolates, 714 were identified correctly, zero were misidentified, zero were unidentified, and six were not characterized as a Gram-positive organism by the VITEK 2 GP method. Additionally, 120 strains exclusive of Gram-positive organisms were screened by Gram stain. A total of 106 isolates were correctly excluded. Fourteen organisms were incorrectly characterized by Gram stain procedures, thus resulting in improper analysis and misidentification by VITEK GP. The VITEK 2 GP identification method is an acceptable automated method for the rapid identification of selected Gram-positive bacteria. PMID:23175976

Crowley, Erin; Bird, Patrick; Fisher, Kiel; Goetz, Katherine; Boyle, Megan; Benzinger, M Joseph; Juenger, Marc; Agin, James; Goins, David; Johnson, Ronald L

2012-01-01

57

Disinfection of gram-negative and gram-positive bacteria using D ynaJ ets® hydrodynamic cavitating jets  

Microsoft Academic Search

Cavitating jet technologies (DynaJets®) were investigated as a means of disinfection of gram-negative Escherichia coli, Klebsiellapneumoniae, Pseudomonas syringae, and Pseudomonas aeruginosa, and gram-positive Bacillus subtilis. The hydrodynamic cavitating jets were found to be very effective in reducing the concentrations of all of these species. In general, the observed rates of disinfection of gram-negative species were higher than for gram-positive species.

Gregory Loraine; Georges Chahine; Chao-Tsung Hsiao; Jin-Keun Choi; Patrick Aley

58

Type IV pili in Gram-positive bacteria.  

PubMed

Type IV pili (T4P) are surface-exposed fibers that mediate many functions in bacteria, including locomotion, adherence to host cells, DNA uptake (competence), and protein secretion and that can act as nanowires carrying electric current. T4P are composed of a polymerized protein, pilin, and their assembly apparatuses share protein homologs with type II secretion systems in eubacteria and the flagella of archaea. T4P are found throughout Gram-negative bacterial families and have been studied most extensively in certain model Gram-negative species. Recently, it was discovered that T4P systems are also widespread among Gram-positive species, in particular the clostridia. Since Gram-positive and Gram-negative bacteria have many differences in cell wall architecture and other features, it is remarkable how similar the T4P core proteins are between these organisms, yet there are many key and interesting differences to be found as well. In this review, we compare the two T4P systems and identify and discuss the features they have in common and where they differ to provide a very broad-based view of T4P systems across all eubacterial species. PMID:24006467

Melville, Stephen; Craig, Lisa

2013-09-01

59

Cystic neutrophilic granulomatous mastitis: an underappreciated pattern strongly associated with gram-positive bacilli.  

PubMed

Although granulomatous lobular mastitis is associated with gram-positive bacilli such as Corynebacterium, this association is not well known. We report 3 cases of mastitis caused by gram-positive bacilli. All 3 abscesses were suppurative with distinct enlarged cystic spaces in which rare gram-positive bacilli were identified. Two cases were also granulomatous. Cultures in all 3 cases were negative. All 3 patients recovered after biopsy and tetracycline-based therapy. Infection in the breast by gram-positive bacilli is associated with a distinct histologic pattern, including cystic spaces in the setting of neutrophilic/granulomatous inflammation that can be recognized and should prompt careful search for the organism within enlarged vacuoles. PMID:21846918

Renshaw, Andrew A; Derhagopian, Robert P; Gould, Edwin W

2011-09-01

60

Vancomycin-resistant Gram-positive bacterial endophthalmitis: epidemiology, treatment options, and outcomes  

PubMed Central

Background The purpose of this study is to evaluate the microbiological profile and treatment outcomes of vancomycin-resistant Gram-positive bacterial endophthalmitis. Medical records of all patients with Gram-positive bacterial endophthalmitis resistant to vancomycin presenting between 1 January 2005 and 31 December 2010 were reviewed in this noncomparative, consecutive, retrospective case series. Favorable outcome was defined as a best-corrected visual acuity of ?20/200. Results Out of 682 culture-positive endophthalmitis isolates, 448/682 (65.6%) were associated with Gram-positive bacteria. In vitro resistance to vancomycin was noted in 7/448 (1.56%). Three cases were posttraumatic, three were postoperative, and one was endogenous in origin. Four Bacillus isolates, two Staphylococcus isolates, and an Enterococcus isolate were resistant. Isolates resistant to vancomycin were sensitive in vitro to ciprofloxacin in 6/7 (86%) patients. Presenting visual acuity was light perception in all seven cases. Favorable outcome was achieved in only 1/7 (14.3%) cases. Conclusions Vancomycin-resistant endophthalmitis is uncommon and usually associated with poor visual outcome. Bacillus sp. is the most frequent Gram-positive bacteria resistant to vancomycin. Fluoroquinolones like ciprofloxacin may be considered as a useful alternative in vancomycin-resistant endophthalmitis.

2013-01-01

61

Longitudinal Assessment of Antimicrobial Susceptibility among Gram-Negative and Gram-Positive Organisms Collected from Italy as Part of the Tigecycline Evaluation and Surveillance Trial between 2004 and 2011  

PubMed Central

The Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) was initiated in 2004 to longitudinally monitor the activity of the broad-spectrum glycylcycline antimicrobial tigecycline, and a suite of comparator agents, against an array of clinically important bacterial pathogens worldwide. In this report, we examine the activity of tigecycline and comparators against a collection of 13,245 clinical isolates, both Gram-positive (n = 4,078 and Gram-negative (n = 9,167), collected from 27 centres in Italy between 2004 and 2011. Susceptibility was established according to Clinical Laboratory Standards Institute guidelines. Tigecycline and linezolid exhibited very good activity against Gram-positive pathogens, with MIC90s ranging from 0.06 to 0.25 mg/L and 1–4 mg/L, respectively; vancomycin and the carbapenems also showed good activity against select Gram-positive pathogens. Tigecycline was the most active agent against Gram-negative pathogens (except P. aeruginosa), with MIC90s ranging from 0.25–2 mg/L (16 mg/L for P. aeruginosa). Amikacin and the carbapenems also possessed good activity against many Gram-negative pathogens here. ESBL-positive E. coli increased in prevalence from 2004 to 2011, while ESBL-positive Klebsiella spp., vancomycin-resistant enterococci and MRSA decreased in prevalence. Linezolid, tigecycline and vancomycin susceptibility were very stable over the course of this study, while susceptibility to ampicillin, piperacillin-tazobactam, ceftriaxone and levofloxacin varied over time according to pathogen; minocycline and cefepime susceptibility among several pathogens decreased during this study.

Stefani, Stefania; Dowzicky, Michael J.

2013-01-01

62

Longitudinal Assessment of Antimicrobial Susceptibility among Gram-Negative and Gram-Positive Organisms Collected from Italy as Part of the Tigecycline Evaluation and Surveillance Trial between 2004 and 2011.  

PubMed

The Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) was initiated in 2004 to longitudinally monitor the activity of the broad-spectrum glycylcycline antimicrobial tigecycline, and a suite of comparator agents, against an array of clinically important bacterial pathogens worldwide. In this report, we examine the activity of tigecycline and comparators against a collection of 13,245 clinical isolates, both Gram-positive (n = 4,078 and Gram-negative (n = 9,167), collected from 27 centres in Italy between 2004 and 2011. Susceptibility was established according to Clinical Laboratory Standards Institute guidelines. Tigecycline and linezolid exhibited very good activity against Gram-positive pathogens, with MIC90s ranging from 0.06 to 0.25 mg/L and 1-4 mg/L, respectively; vancomycin and the carbapenems also showed good activity against select Gram-positive pathogens. Tigecycline was the most active agent against Gram-negative pathogens (except P. aeruginosa), with MIC90s ranging from 0.25-2 mg/L (16 mg/L for P. aeruginosa). Amikacin and the carbapenems also possessed good activity against many Gram-negative pathogens here. ESBL-positive E. coli increased in prevalence from 2004 to 2011, while ESBL-positive Klebsiella spp., vancomycin-resistant enterococci and MRSA decreased in prevalence. Linezolid, tigecycline and vancomycin susceptibility were very stable over the course of this study, while susceptibility to ampicillin, piperacillin-tazobactam, ceftriaxone and levofloxacin varied over time according to pathogen; minocycline and cefepime susceptibility among several pathogens decreased during this study. PMID:24287463

Stefani, Stefania; Dowzicky, Michael J

2013-01-01

63

Regulation of Apoptosis by Gram-Positive Bacteria  

PubMed Central

Apoptosis, or programmed cell death (PCD), is an important physiological mechanism, through which the human immune system regulates homeostasis and responds to diverse forms of cellular damage. PCD may also be involved in immune counteraction to microbial infection. Over the past decade, the amount of research on bacteria-induced PCD has grown tremendously, and the implications of this mechanism on immunity are being elucidated. Some pathogenic bacteria actively trigger the suicide response in critical lineages of leukocytes that orchestrate both the innate and adaptive immune responses; other bacteria proactively prevent PCD to benefit their own survival and persistence. Currently, the microbial virulence factors, which represent the keys to unlocking the suicide response in host cells, are a primary focus of this field. In this review, we discuss these bacterial “apoptosis regulatory molecules” and the apoptotic events they either trigger or prevent, the host target cells of this regulatory activity, and the possible ramifications for immunity to infection. Gram-positive pathogens including Staphylococcus, Streptococcus, Bacillus, Listeria, and Clostridia species are discussed as important agents of human infection that modulate PCD pathways in eukaryotic cells.

Ulett, Glen C.; Adderson, Elisabeth E.

2008-01-01

64

Dry and moist heat sterilisation cannot inactivate pyrogenicity of Gram positive microorganisms  

Microsoft Academic Search

In the monocytic cell line Mono Mac 6 pyrogens induce interleukin-6 secretion dose dependently. The aim of this study is to examine the interleukin-6 inducing capacity of Gram positive Staphylococcus aureus and Bacillus subtilis endospores after moist and dry heat sterilisation. Moist heat sterilisation of B. subtilis endospores for 15min at 121°C and 134°C can only reduce the interleukin-6 inducing

Lise Moesby; Erik W. Hansen; Jens D. Christensen; Catrine H. Høyer; Gitte L. Juhl; Helle B. Olsen

2005-01-01

65

Multidrug resistance in hydrocarbon-tolerant Gram-positive and Gram-negative bacteria.  

PubMed

New Gram-positive and Gram-negative bacteria were isolated from Poeni oily sludge, using enrichment procedures. The six Gram-positive strains belong to Bacillus, Lysinibacillus and Rhodococcus genera. The eight Gram-negative strains belong to Shewanella, Aeromonas, Pseudomonas and Klebsiella genera. Isolated bacterial strains were tolerant to saturated (i.e., n-hexane, n-heptane, n-decane, n-pentadecane, n-hexadecane, cyclohexane), monoaromatic (i.e., benzene, toluene, styrene, xylene isomers, ethylbenzene, propylbenzene) and polyaromatic (i.e., naphthalene, 2-methylnaphthalene, fluorene) hydrocarbons, and also resistant to different antimicrobial agents (i.e., ampicillin, kanamycin, rhodamine 6G, crystal violet, malachite green, sodium dodecyl sulfate). The presence of hydrophilic antibiotics like ampicillin or kanamycin in liquid LB-Mg medium has no effects on Gram-positive and Gram-negative bacteria resistance to toxic compounds. The results indicated that Gram-negative bacteria are less sensitive to toxic compounds than Gram-positive bacteria, except one bacteria belonging to Lysinibacillus genus. There were observed cellular and molecular modifications induced by ampicillin or kanamycin to isolated bacterial strains. Gram-negative bacteria possessed between two and four catabolic genes (alkB, alkM, alkB/alkB1, todC1, xylM, PAH dioxygenase, catechol 2,3-dioxygenase), compared with Gram-positive bacteria (except one bacteria belonging to Bacillus genus) which possessed one catabolic gene (alkB/alkB1). Transporter genes (HAE1, acrAB) were detected only in Gram-negative bacteria. PMID:21478643

Stancu, Mihaela Marilena; Grifoll, Magdalena

2011-01-01

66

Distinct localization of the complement C5b-9 complex on Gram-positive bacteria.  

PubMed

The plasma proteins of the complement system fulfil important immune defence functions, including opsonization of bacteria for phagocytosis, generation of chemo-attractants and direct bacterial killing via the Membrane Attack Complex (MAC or C5b-9). The MAC is comprised of C5b, C6, C7, C8, and multiple copies of C9 that generate lytic pores in cellular membranes. Gram-positive bacteria are protected from MAC-dependent lysis by their thick peptidoglycan layer. Paradoxically, several Gram-positive pathogens secrete small proteins that inhibit C5b-9 formation. In this study, we found that complement activation on Gram-positive bacteria in serum results in specific surface deposition of C5b-9 complexes. Immunoblotting revealed that C9 occurs in both monomeric and polymeric (SDS-stable) forms, indicating the presence of ring-structured C5b-9. Surprisingly, confocal microscopy demonstrated that C5b-9 deposition occurs at specialized regions on the bacterial cell. On Streptococcus pyogenes, C5b-9 deposits near the division septum whereas on Bacillus subtilis the complex is located at the poles. This is in contrast to C3b deposition, which occurs randomly on the bacterial surface. Altogether, these results show a previously unrecognized interaction between the C5b-9 complex and Gram-positive bacteria, which might ultimately lead to a new model of MAC assembly and functioning. PMID:23869880

Berends, Evelien T M; Dekkers, Johanna F; Nijland, Reindert; Kuipers, Annemarie; Soppe, Jasper A; van Strijp, Jos A G; Rooijakkers, Suzan H M

2013-12-01

67

An extreme-halophile archaebacterium possesses the interlock type of prephenate dehydratase characteristic of the Gram-positive eubacteria  

NASA Technical Reports Server (NTRS)

The focal point of phenylalanine biosynthesis is a dehydratase reaction which in different organisms may be prephenate dehydratase, arogenate dehydratase, or cyclohexadienyl dehydratase. Gram-positive, Gram-negative, and cyanobacterial divisions of the eubacterial kingdom exhibit different dehydratase patterns. A new extreme-halophile isolate, which grows on defined medium and is tentatively designated as Halobacterium vallismortis CH-1, possesses the interlock type of prephenate dehydratase present in Gram-positive bacteria. In addition to the conventional sensitivity to feedback inhibition by L-phenylalanine, the phenomenon of metabolic interlock was exemplified by the sensitivity of prephenate dehydratase to allosteric effects produced by extra-pathway (remote) effectors. Thus, L-tryptophan inhibited activity while L-tyrosine, L-methionine, L-leucine and L-isoleucine activated the enzyme. L-Isoleucine and L-phenylalanine were effective at micromolar levels; other effectors operated at mM levels. A regulatory mutant selected for resistance to growth inhibition caused by beta-2-thienylalanine possessed an altered prephenate dehydratase in which a phenomenon of disproportionately low activity at low enzyme concentration was abolished. Inhibition by L-tryptophan was also lost, and activation by allosteric activators was diminished. Not only was sensitivity to feedback inhibition by L-phenylalanine lost, but the mutant enzyme was now activated by this amino acid (a mutation type previously observed in Bacillus subtilis). It remains to be seen whether this type of prephenate dehydratase will prove to be characteristic of all archaebacteria or of some archaebacterial subgroup cluster.

Jensen, R. A.; d'Amato, T. A.; Hochstein, L. I.

1988-01-01

68

Multiplex Identification of Gram-Positive Bacteria and Resistance Determinants Directly from Positive Blood Culture Broths: Evaluation of an Automated Microarray-Based Nucleic Acid Test  

PubMed Central

Background A multicenter study was conducted to evaluate the diagnostic accuracy (sensitivity and specificity) of the Verigene Gram-Positive Blood Culture Test (BC-GP) test to identify 12 Gram-positive bacterial gene targets and three genetic resistance determinants directly from positive blood culture broths containing Gram-positive bacteria. Methods and Findings 1,252 blood cultures containing Gram-positive bacteria were prospectively collected and tested at five clinical centers between April, 2011 and January, 2012. An additional 387 contrived blood cultures containing uncommon targets (e.g., Listeria spp., S. lugdunensis, vanB-positive Enterococci) were included to fully evaluate the performance of the BC-GP test. Sensitivity and specificity for the 12 specific genus or species targets identified by the BC-GP test ranged from 92.6%–100% and 95.4%–100%, respectively. Identification of the mecA gene in 599 cultures containing S. aureus or S. epidermidis was 98.6% sensitive and 94.3% specific compared to cefoxitin disk method. Identification of the vanA gene in 81 cultures containing Enterococcus faecium or E. faecalis was 100% sensitive and specific. Approximately 7.5% (87/1,157) of single-organism cultures contained Gram-positive bacteria not present on the BC-GP test panel. In 95 cultures containing multiple organisms the BC-GP test was in 71.6% (68/95) agreement with culture results. Retrospective analysis of 107 separate blood cultures demonstrated that identification of methicillin resistant S. aureus and vancomycin resistant Enterococcus spp. was completed an average of 41.8 to 42.4 h earlier using the BC-GP test compared to routine culture methods. The BC-GP test was unable to assign mecA to a specific organism in cultures containing more than one Staphylococcus isolate and does not identify common blood culture contaminants such as Micrococcus, Corynebacterium, and Bacillus. Conclusions The BC-GP test is a multiplex test capable of detecting most leading causes of Gram-positive bacterial blood stream infections as well as genetic markers of methicillin and vancomycin resistance directly from positive blood cultures. Please see later in the article for the Editors' Summary

Buchan, Blake W.; Ginocchio, Christine C.; Manii, Ryhana; Cavagnolo, Robert; Pancholi, Preeti; Swyers, Lettie; Thomson, Richard B.; Anderson, Christopher; Kaul, Karen; Ledeboer, Nathan A.

2013-01-01

69

Chocolate agar, a differential medium for gram-positive cocci.  

PubMed Central

Reactions incurred on chocolate agar by gram-positive cocci were correlated with species identity. Darkening and clearing of the medium was usually associated with the species Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus simulans, and Streptococcus faecalis. Yellowing of chocolate agar was associated with alpha-hemolytic species of Streptococcus. The study demonstrated that reactions occurring on chocolate agar are useful in identifying gram-positive cocci.

Gunn, B A

1984-01-01

70

Moderately halophilic gram-positive bacterial diversity in hypersaline environments  

Microsoft Academic Search

Moderately halophilic bacteria are microorganisms that grow optimally in media containing 3%–15% (w\\/v) salt. They are represented\\u000a by a heterogeneous group of microorganisms included in many different genera. Gram-negative moderately halophilic bacteria\\u000a have been studied in more detail, but studies on gram-positive species are more scarce. Recent studies carried out by our\\u000a research group on gram-positive moderate halophiles have permitted

Antonio Ventosa; M. Carmen Márquez; María J. Garabito; David R. Arahal

1998-01-01

71

Telavancin: a new lipoglycopeptide for gram-positive infections.  

PubMed

Telavancin is a lipoglycopeptide derivative of vancomycin. Similar to vancomycin, it demonstrates activity in vitro against a variety of Gram-positive pathogens, including but not limited to methicillin-resistant Staphylococccus aureus (MRSA) and penicillin-resistant Streptococcus pneumoniae (PRSP). Modifications to vancomycin's structure expanded telavancin's spectrum of activity in vitro to include organisms such as glycopeptide-intermediate S. aureus (GISA), vancomycin-resistant S. aureus (VRSA) and vancomycin-resistant enterococci (VRE). However, the clinical implications of this are currently unknown. Similar to other glycopeptides, televancin binds to the D-alanyl-D-alanine (D-Ala-D-Ala) terminus in Gram-positive organisms, resulting in inhibition of bacterial cell wall synthesis. In addition, telavancin causes depolarization of the bacterial cell membrane and increased membrane permeability. The resulting activity in vitro is rapidly bactericidal and concentration dependent, with the ratio of area under the time concentration curve to minimum inhibitory concentration (AUC/MIC) as the best predictor of activity in animal models to date. In humans, telavancin exhibits a pharmacokinetic profile that permits once-daily intravenous administration. Doses of 7.5 and 10 mg/kg/day have been studied in clinical trials. The need for dosage adjustments based on age, gender and obesity appear unnecessary. In addition, moderate hepatic impairment does not appreciably alter the pharmacokinetics of the drug. Because telavancin is extensively cleared by the kidneys, dosage adjustments will be required in patients with moderate to severe renal impairment. Published phase II and III clinical trials have shown telavancin to be comparable to standard therapy for the treatment of complicated skin and soft tissue infections. Clinical trials in the treatment of S. aureus bacteremia and hospital-acquired pneumonia are under way. Adverse effects overall appear to be mild and reversible, with taste disturbance, foamy urine, headache, procedural site pain, nausea and vomiting being the most commonly reported. However, renal toxicity was reported more frequently with telavancin than with vancomycin in two phase III clinical trials (3% versus 1%). Prolongation of the corrected QT (QTc) interval has been more common with telavancin than comparator agents, but no clinically significant electrocardiogram (ECG) changes or cardiac abnormalities have been observed to date. Although human pregnancy data is not currently available, animal data revealed limb malformations that were possibly related to telavancin therapy. Therefore, the potential teratogenicity of this agent must be considered in women who are pregnant or may become pregnant. PMID:19436839

Smith, Winter J; Drew, Richard H

2009-03-01

72

Enhancement of Growth of Aerobic, Anaerobic, and Facultative Bacteria in Mixed Infections with Anaerobic and Facultative Gram-Positive Cocci,  

National Technical Information Service (NTIS)

Anaerobic and facultative gram-positive cocci (AFGPC) mixed with other bacteria are frequently recovered from infections at different sites in the body. Although many of these organisms are synergistic, the exact role of AFGPC in these infections and thei...

I. Brook

1988-01-01

73

Glycopeptide resistance in gram-positive cocci: a review.  

PubMed

Vancomycin-resistant enterococci (VRE) have emerged as important nosocomial pathogens in the past two decades all over the world and have seriously limited the choices available to clinicians for treating infections caused by these agents. Methicillin-resistant Staphylococcus aureus, perhaps the most notorious among the nosocomial pathogens, was till recently susceptible to vancomycin and the other glycopeptides. Emergence of vancomycin nonsusceptible strains of S. aureus has led to a worrisome scenario where the options available for treating serious infections due to these organisms are very limited and not well evaluated. Vancomycin resistance in clinically significant isolates of coagulase-negative staphylococci is also on the rise in many setups. This paper aims to highlight the genetic basis of vancomycin resistance in Enterococcus species and S. aureus. It also focuses on important considerations in detection of vancomycin resistance in these gram-positive bacteria. The problem of glycopeptide resistance in clinical isolates of coagulase-negative staphylococci and the phenomenon of vancomycin tolerance seen in some strains of Streptococcus pneumoniae has also been discussed. Finally, therapeutic options available and being developed against these pathogens have also found a mention. PMID:22778729

Sujatha, S; Praharaj, Ira

2012-01-01

74

Metabolic versatility of Gram-positive microbial isolates from contaminated river sediments.  

PubMed

Gram-positive bacteria from river sediments affected by the proximity of a petrochemical industrial site were isolated and characterized with respect to their ability to degrade a wide range of aromatic compounds. In this study we identified metabolically diverse Gram-positive bacteria capable of growth on wide range aromatic compounds in the presence of heavy metals and with the ability to accumulate biopolymers. Thirty-four isolates that were able to use 9 or more common aromatic pollutants, such as benzene, biphenyl, naphthalene etc. as a sole source of carbon and energy included members of Bacillus, Arthrobacter, Rhodococcus, Gordonia, Streptomyces, and Staphylococcus genus. Rhodococcus sp. TN105, Gordonia sp. TN103 and Arthrobacter sp. TN221 were identified as novel strains. Nine isolates were able to grow in the presence of one or more metals (mercury, cadmium, nickel) at high concentration (100mM). Seven isolates could degrade 15 different aromatic compounds and could grow in the presence of one or more heavy metals. Two of these isolates were resistant to multiple antibiotics including erythromycin and nalidixic acid. One third of isolates could accumulate at least one biopolymer. Twelve isolates (mainly Bacillus sp. and Arthrobacter sp.) accumulated polyphosphate, 3 Bacillus sp. accumulated polyhydroxybutyrate, while 4 isolates could accumulate exopolysaccharides. PMID:22421345

Narancic, Tanja; Djokic, Lidija; Kenny, Shane T; O'Connor, Kevin E; Radulovic, Vanja; Nikodinovic-Runic, Jasmina; Vasiljevic, Branka

2012-05-15

75

Biosynthesis of Auxin by the Gram-Positive Phytopathogen Rhodococcus fascians Is Controlled by Compounds Specific to Infected Plant Tissues  

Microsoft Academic Search

The role and metabolism of indole-3-acetic acid in gram-negative bacteria is well documented, but little is known about indole-3-acetic acid biosynthesis and regulation in gram-positive bacteria. The phytopathogen Rhodococcus fascians, a gram-positive organism, incites diverse developmental alterations, such as leafy galls, on a wide range of plants. Phenotypic analysis of a leafy gall suggests that auxin may play an important

Olivier Vandeputte; Sevgi Oden; Adeline Mol; Danny Vereecke; Koen Goethals; Mondher El Jaziri; Els Prinsen

2005-01-01

76

Protein secretion and surface display in Gram-positive bacteria  

PubMed Central

The cell wall peptidoglycan of Gram-positive bacteria functions as a surface organelle for the transport and assembly of proteins that interact with the environment, in particular, the tissues of an infected host. Signal peptide-bearing precursor proteins are secreted across the plasma membrane of Gram-positive bacteria. Some precursors carry C-terminal sorting signals with unique sequence motifs that are cleaved by sortase enzymes and linked to the cell wall peptidoglycan of vegetative forms or spores. The sorting signals of pilin precursors are cleaved by pilus-specific sortases, which generate covalent bonds between proteins leading to the assembly of fimbrial structures. Other precursors harbour surface (S)-layer homology domains (SLH), which fold into a three-pronged spindle structure and bind secondary cell wall polysaccharides, thereby associating with the surface of specific Gram-positive microbes. Type VII secretion is a non-canonical secretion pathway for WXG100 family proteins in mycobacteria. Gram-positive bacteria also secrete WXG100 proteins and carry unique genes that either contribute to discrete steps in secretion or represent distinctive substrates for protein transport reactions.

Schneewind, Olaf; Missiakas, Dominique M.

2012-01-01

77

Novel antimicrobial peptides that inhibit gram positive bacterial exotoxin synthesis.  

PubMed

Gram-positive bacteria, such as Staphylococcus aureus, cause serious human illnesses through combinations of surface virulence factors and secretion of exotoxins. Our prior studies using the protein synthesis inhibitor clindamycin and signal transduction inhibitors glycerol monolaurate and ?-globin and ?-globin chains of hemoglobin indicate that their abilities to inhibit exotoxin production by S. aureus are separable from abilities to inhibit growth of the organism. Additionally, our previous studies suggest that inhibition of exotoxin production, in absence of ability to kill S. aureus and normal flora lactobacilli, will prevent colonization by pathogenic S. aureus, while not interfering with lactobacilli colonization. These disparate activities may be important in development of novel anti-infective agents that do not alter normal flora. We initiated studies to explore the exotoxin-synthesis-inhibition activity of hemoglobin peptides further to develop potential agents to prevent S. aureus infections. We tested synthesized ?-globin chain peptides, synthetic variants of ?-globin chain peptides, and two human defensins for ability to inhibit exotoxin production without significantly inhibiting S. aureus growth. All of these peptides were weakly or not inhibitory to bacterial growth. However, the peptides were inhibitory to exotoxin production with increasing activity dependent on increasing numbers of positively-charged amino acids. Additionally, the peptides could be immobilized on agarose beads or have amino acid sequences scrambled and still retain exotoxin-synthesis-inhibition. The peptides are not toxic to human vaginal epithelial cells and do not inhibit growth of normal flora L. crispatus. These peptides may interfere with plasma membrane signal transduction in S. aureus due to their positive charges. PMID:24748386

Merriman, Joseph A; Nemeth, Kimberly A; Schlievert, Patrick M

2014-01-01

78

Conjugative Plasmid Transfer in Gram-Positive Bacteria  

PubMed Central

Conjugative transfer of bacterial plasmids is the most efficient way of horizontal gene spread, and it is therefore considered one of the major reasons for the increase in the number of bacteria exhibiting multiple-antibiotic resistance. Thus, conjugation and spread of antibiotic resistance represents a severe problem in antibiotic treatment, especially of immunosuppressed patients and in intensive care units. While conjugation in gram-negative bacteria has been studied in great detail over the last decades, the transfer mechanisms of antibiotic resistance plasmids in gram-positive bacteria remained obscure. In the last few years, the entire nucleotide sequences of several large conjugative plasmids from gram-positive bacteria have been determined. Sequence analyses and data bank comparisons of their putative transfer (tra) regions have revealed significant similarities to tra regions of plasmids from gram-negative bacteria with regard to the respective DNA relaxases and their targets, the origins of transfer (oriT), and putative nucleoside triphosphatases NTP-ases with homologies to type IV secretion systems. In contrast, a single gene encoding a septal DNA translocator protein is involved in plasmid transfer between micelle-forming streptomycetes. Based on these clues, we propose the existence of two fundamentally different plasmid-mediated conjugative mechanisms in gram-positive microorganisms, namely, the mechanism taking place in unicellular gram-positive bacteria, which is functionally similar to that in gram-negative bacteria, and a second type that occurs in multicellular gram-positive bacteria, which seems to be characterized by double-stranded DNA transfer.

Grohmann, Elisabeth; Muth, Gunther; Espinosa, Manuel

2003-01-01

79

Comparative in vitro activity of levofloxacin and ofloxacin against Gram-positive bacteria  

Microsoft Academic Search

The in vitro activity of levofloxacin against 506 Gram-positive bacteria was compared with those of D(?)-ofloxacin, ofloxacin, ciprofloxacin, and sparfloxacin. Levofloxacin was generally twice as active as ofloxacin against these organisms (range, 0–3 twofold dilutions). Sparfloxacin appeared to have the greatest activity overall, but for several groups of organisms minimum inhibitory concentrations (MIC90s) of this com pound were within one

George M. Eliopoulos; Christine B. Wennersten; Robert C. Moellering

1996-01-01

80

The opening of the SPP1 bacteriophage tail, a prevalent mechanism in Gram-positive-infecting siphophages.  

PubMed

The SPP1 siphophage uses its long non-contractile tail and tail tip to recognize and infect the Gram-positive bacterium Bacillus subtilis. The tail-end cap and its attached tip are the critical components for host recognition and opening of the tail tube for genome exit. In the present work, we determined the cryo-electron microscopic (cryo-EM) structure of a complex formed by the cap protein gp19.1 (Dit) and the N terminus of the downstream protein of gp19.1 in the SPP1 genome, gp21(1-552) (Tal). This complex assembles two back-to-back stacked gp19.1 ring hexamers, interacting loosely, and two gp21(1-552) trimers interacting with gp19.1 at both ends of the stack. Remarkably, one gp21(1-552) trimer displays a "closed" conformation, whereas the second is "open" delineating a central channel. The two conformational states dock nicely into the EM map of the SPP1 cap domain, respectively, before and after DNA release. Moreover, the open/closed conformations of gp19.1-gp21(1-552) are consistent with the structures of the corresponding proteins in the siphophage p2 baseplate, where the Tal protein (ORF16) attached to the ring of Dit (ORF15) was also found to adopt these two conformations. Therefore, the present contribution allowed us to revisit the SPP1 tail distal-end architectural organization. Considering the sequence conservation among Dit and the N-terminal region of Tal-like proteins in Gram-positive-infecting Siphoviridae, it also reveals the Tal opening mechanism as a hallmark of siphophages probably involved in the generation of the firing signal initiating the cascade of events that lead to phage DNA release in vivo. PMID:21622577

Goulet, Adeline; Lai-Kee-Him, Joséphine; Veesler, David; Auzat, Isabelle; Robin, Gautier; Shepherd, Dale A; Ashcroft, Alison E; Richard, Eric; Lichière, Julie; Tavares, Paulo; Cambillau, Christian; Bron, Patrick

2011-07-15

81

Vancomycin-Resistant Gram-Positive Cocci Isolated from the Saliva of Wild Songbirds  

PubMed Central

We analyzed highly vancomycin-resistant Gram-positive bacteria isolated from the saliva of migratory songbirds captured, sampled, and released from a birdbanding station in western Kansas. Individual bacterial isolates were identified by partial 16S rRNA sequencing. Most of the bacteria in this study were shown to be Staphylococcus succinus with the majority being isolated from the American Robin. Some of these bacteria were shown to carry vanA, vanB, and vanC vancomycin-resistance genes and have the ability to form biofilms. One of the van gene-carrying isolates is also coagulase positive, which is normally considered a virulence factor. Other organisms isolated included Staphylococcus saprophyticus as well as Enterococcus gallinarum. Given the wide range of the American Robin and ease of horizontal gene transfer between Gram-positive cocci, we postulate that these organisms could serve as a reservoir of vancomycin-resistance genes capable of transferring to human pathogens.

Ishihara, Shingo; Bitner, Jessica J.; Farley, Greg H.

2014-01-01

82

Which antibiotic for resistant Gram-positives, and why?  

PubMed

Increasing resistance in Gram-positive pathogens, particularly Staphylococcus aureus, and enterococcus, has become a major clinical problem, particularly in the hospital environment, causing significant morbidity and mortality in both healthy hosts and in those with underlying comorbidities. Increased resistance drives the use of empiric therapy with less well-studied and potentially more toxic agents. Resistance mechanisms for currently recommended agents are discussed, with options for therapy of resistant pathogens. For any new agent used, resistance is likely to develop, which underscores the concept that both antibiotics and antimicrobial resistance are ancient, and only by prudent use of antimicrobial agents and effective infection control measures when resistance arises, will effective agents be available to treat Gram-positive pathogens in the future. PMID:24188585

Bradley, John S

2014-01-01

83

Progress in the Development of Effective Vaccines to Prevent Selected Gram Positive Bacterial Infections  

PubMed Central

Infections due to virulent gram positive bacteria, such as Staphylococcus aureus, group B streptococci and group A streptococci remain significant causes of morbidity and mortality despite progress in antimicrobial therapy. Despite significant advances in the understanding of the pathogenesis of infection due to these organisms, there are only limited strategies to prevent infection. In this paper, we review efforts to develop safe and effective vaccines that would prevent infections due to these 3 pathogens.

Bronze, Michael S.; Dale, James B.

2010-01-01

84

Conjugative type IV secretion systems in Gram-positive bacteria  

PubMed Central

Bacterial conjugation presents the most important means to spread antibiotic resistance and virulence factors among closely and distantly related bacteria. Conjugative plasmids are the mobile genetic elements mainly responsible for this task. All the genetic information required for the horizontal transmission is encoded on the conjugative plasmids themselves. Two distinct concepts for horizontal plasmid transfer in Gram-positive bacteria exist, the most prominent one transports single stranded plasmid DNA via a multi-protein complex, termed type IV secretion system, across the Gram-positive cell envelope. Type IV secretion systems have been found in virtually all unicellular Gram-positive bacteria, whereas multicellular Streptomycetes seem to have developed a specialized system more closely related to the machinery involved in bacterial cell division and sporulation, which transports double stranded DNA from donor to recipient cells. This review intends to summarize the state of the art of prototype systems belonging to the two distinct concepts; it focuses on protein key players identified so far and gives future directions for research in this emerging field of promiscuous interbacterial transport.

Goessweiner-Mohr, Nikolaus; Arends, Karsten; Keller, Walter; Grohmann, Elisabeth

2013-01-01

85

Update on antimicrobial susceptibility rates among gram-negative and gram-positive organisms in the United States: Results from the Tigecycline Evaluation and Surveillance Trial (TEST) 2005 to 2007  

Microsoft Academic Search

Background: The Tigecycline Evaluation and Surveillance Trial (TTEST) is a global surveillance study initiated in 2004.Its goal is to assess the in vitro activity of the glycylcycline, tigecycline, and comparator antimicrobials.Objective: The aim of this study was to measure the in vitro activity of a panel of antimicrobial agents against gram-nnegative and gram-ppositive organisms collected in the United States in

Michael J. Dowzicky; Choong H. Park

2008-01-01

86

Heterologous Protein Secretion by Bacillus Species  

Microsoft Academic Search

The Gram-positive bacterium Bacillus subtilis and some of its close relatives are widely used for the industrial production of enzymes for the detergents, food, and beverage industries. The choice of these organisms is based almost exclusively on the high capacity of their secretion systems that are, under the right conditions, able to secrete proteins at grams per liter concentrations. In

Susanne Pohl; Colin R. Harwood

2010-01-01

87

Complete Genome of Bacillus megaterium Podophage Page.  

PubMed

Bacillus megaterium is a Gram-positive, spore-forming saprophytic inhabitant of diverse environments. It is a reservoir for industrial chemical production and is emerging as a model organism for studying sporulation and protein localization. Here, we introduce the complete genome of Page, a novel podophage infecting B. megaterium. PMID:24744341

Lopez, Mariana S; Hodde, Mary K; Chamakura, Karthik R; Kuty Everett, Gabriel F

2014-01-01

88

Complete Genome of Bacillus subtilis Myophage Grass  

PubMed Central

Bacillus subtilis is a ubiquitous Gram-positive model organism. Here, we describe the complete genome of B. subtilus myophage Grass. Aside from genes encoding core proteins pertinent to the life cycle of the phage, Grass has several interesting features, including an FtsK/SpoIIIE protein.

Miller, Stanton Y.; Colquhoun, Jennifer M.; Perl, Abbey L.; Chamakura, Karthik R.

2013-01-01

89

Evaluation of the Nanosphere Verigene Gram-Positive Blood Culture Assay with the VersaTREK Blood Culture System and Assessment of Possible Impact on Selected Patients  

PubMed Central

The Verigene Gram-positive blood culture (BC-GP) assay (Nanosphere, Northbrook, IL) is a molecular method for the rapid identification of Gram-positive organisms and resistance markers directly from blood culture bottles. A total of 148 VersaTREK REDOX 1 40-ml aerobic bottles demonstrating Gram-positive bacteria were tested. Results were compared with those from conventional biochemical and matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) identifications. We obtained isolates of methicillin-resistant Staphylococcus aureus (MRSA) (24), methicillin-susceptible Staphylococcus aureus (MSSA) (14), methicillin-resistant Staphylococcus epidermidis (MRSE) (17), methicillin-susceptible Staphylococcus epidermidis (MSSE) (9), other coagulase-negative staphylococci (19), Streptococcus salivarius (5), Streptococcus parasanguinis (2), Streptococcus sanguinis (1), Streptococcus cristatus (1), the Streptococcus bovis group (5), Streptococcus agalactiae (9), the Streptococcus anginosus group (1), Streptococcus pneumoniae (6), vancomycin-resistant Enterococcus faecium (VRE FCM) (16), vancomycin-susceptible Enterococcus faecalis (3), Aerococcus viridans (2), Bacillus (6), Corynebacterium (8), Lactobacillus (2), Micrococcus (2), Neisseria mucosa (1), Escherichia coli (3), Candida tropicalis (1), Propionibacterium (1), and Rothia (1). Overall agreement with the culture results was 95%. A total of 137 of 138 (99%) monomicrobial cultures were concordant. We tested 9 polymicrobial samples and found 33% agreement. A chart review of 31 patients with MRSA, MSSA, or VRE demonstrated that the Nanosphere BC-GP assay might have led to more appropriate antibiotic selection for these patients an average of 42 h earlier. Additionally, contact isolation could have been initiated an average of 37 h earlier for patients with MRSA or VRE. The BC-GP assay may have a positive impact on patient care, health care costs, and antibiotic stewardship.

Beal, Stacy G.; Ciurca, Jane; Smith, Geremy; John, Jeffrey; Lee, Francesca; Doern, Christopher D.

2013-01-01

90

Evaluation of the nanosphere verigene gram-positive blood culture assay with the VersaTREK blood culture system and assessment of possible impact on selected patients.  

PubMed

The Verigene Gram-positive blood culture (BC-GP) assay (Nanosphere, Northbrook, IL) is a molecular method for the rapid identification of Gram-positive organisms and resistance markers directly from blood culture bottles. A total of 148 VersaTREK REDOX 1 40-ml aerobic bottles demonstrating Gram-positive bacteria were tested. Results were compared with those from conventional biochemical and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) identifications. We obtained isolates of methicillin-resistant Staphylococcus aureus (MRSA) (24), methicillin-susceptible Staphylococcus aureus (MSSA) (14), methicillin-resistant Staphylococcus epidermidis (MRSE) (17), methicillin-susceptible Staphylococcus epidermidis (MSSE) (9), other coagulase-negative staphylococci (19), Streptococcus salivarius (5), Streptococcus parasanguinis (2), Streptococcus sanguinis (1), Streptococcus cristatus (1), the Streptococcus bovis group (5), Streptococcus agalactiae (9), the Streptococcus anginosus group (1), Streptococcus pneumoniae (6), vancomycin-resistant Enterococcus faecium (VRE FCM) (16), vancomycin-susceptible Enterococcus faecalis (3), Aerococcus viridans (2), Bacillus (6), Corynebacterium (8), Lactobacillus (2), Micrococcus (2), Neisseria mucosa (1), Escherichia coli (3), Candida tropicalis (1), Propionibacterium (1), and Rothia (1). Overall agreement with the culture results was 95%. A total of 137 of 138 (99%) monomicrobial cultures were concordant. We tested 9 polymicrobial samples and found 33% agreement. A chart review of 31 patients with MRSA, MSSA, or VRE demonstrated that the Nanosphere BC-GP assay might have led to more appropriate antibiotic selection for these patients an average of 42 h earlier. Additionally, contact isolation could have been initiated an average of 37 h earlier for patients with MRSA or VRE. The BC-GP assay may have a positive impact on patient care, health care costs, and antibiotic stewardship. PMID:24048531

Beal, Stacy G; Ciurca, Jane; Smith, Geremy; John, Jeffrey; Lee, Francesca; Doern, Christopher D; Gander, Rita M

2013-12-01

91

Comparative genomics of the methionine metabolism in Gram-positive bacteria: a variety of regulatory systems.  

PubMed

Regulation of the methionine biosynthesis and transport genes in bacteria is rather diverse and involves two RNA-level regulatory systems and at least three DNA-level systems. In particular, the methionine metabolism in Gram-positive bacteria was known to be controlled by the S-box and T-box mechanisms, both acting on the level of premature termination of transcription. Using comparative analysis of genes, operons and regulatory elements, we described the methionine metabolic pathway and the methionine regulons in available genomes of Gram-positive bacteria. A large number of methionine-specific RNA elements were identified. S-boxes were shown to be widely distributed in Bacillales and Clostridia, whereas methionine-specific T-boxes occurred mostly in Lactobacillales. A candidate binding signal (MET-box) for a hypothetical methionine regulator, possibly MtaR, was identified in Streptococcaceae, the only family in the Bacillus/Clostridium group of Gram-positive bacteria having neither S-boxes, nor methionine-specific T-boxes. Positional analysis of methionine-specific regulatory sites complemented by genome context analysis lead to identification of new members of the methionine regulon, both enzymes and transporters, and reconstruction of the methionine metabolism in various bacterial genomes. In particular, we found candidate transporters for methionine (MetT) and methylthioribose (MtnABC), as well as new enzymes forming the S-adenosylmethionine recycling pathway. Methionine biosynthetic enzymes in various bacterial species are quite variable. In particular, Oceanobacillus iheyensis possibly uses a homolog of the betaine-homocysteine methyltransferase bhmT gene from vertebrates to substitute missing bacterial-type methionine synthases. PMID:15215334

Rodionov, Dmitry A; Vitreschak, Alexey G; Mironov, Andrey A; Gelfand, Mikhail S

2004-01-01

92

Glycerol Monolaurate Inhibits the Effects of Gram Positive Select Agents on Eukaryotic Cells†  

PubMed Central

Many exotoxins of gram positive bacteria, such as superantigens (staphylococcal enterotoxins, toxic shock syndrome toxin-1 [TSST-1], and streptococcal pyrogenic exotoxins) and anthrax toxin are bioterrorism agents that cause diseases by immunostimulation or cytotoxicity. Glycerol monolaurate (GML), a fatty acid monoester found naturally in humans, has been reported to prevent synthesis of gram positive bacterial exotoxins. This study explored the ability of GML to inhibit the effects of exotoxins on mammalian cells and prevent rabbit lethality from TSS. GML (?10 ug/ml) inhibited superantigen (5 ug/ml) immunoproliferation, as determined by inhibition of 3H-thymidine incorporation into DNA of human peripheral blood mononuclear cells (1 × 106 cells/ml) as well as phospholipase C?1, suggesting inhibition of signal transduction. The compound (20 ug/ml) prevented superantigen (100 ug/ml) induced cytokine secretion by human vaginal epithelial cells (HVECs) as measured by ELISA. GML (250 ug) inhibited rabbit lethality due to TSST-1 administered vaginally. GML (10 ug/ml) inhibited HVEC and macrophage cytotoxicity by anthrax toxin, prevented erythrocyte lysis by purified hemolysins (staphylococcal ? and ?) and culture fluids containing streptococcal and Bacillus anthracis hemolysins, and was non-toxic to mammalian cells (up to 100 ug/ml) and rabbits (250 ug). GML stabilized mammalian cell membranes, as erythrocyte lysis was reduced in the presence of hypotonic aqueous solutions (0 to 0.05 M saline) or staphylococcal ? and ?-hemolysins when erythrocytes were pretreated with GML. GML may be useful in management of gram positive exotoxin illnesses; its action appears to be membrane stabilization with inhibition of signal transduction.

Peterson, Marnie L.; Schlievert, Patrick M.

2008-01-01

93

Speciation of Gram-positive bacteria in fresh and ambient-stored sub-tropical marine fish.  

PubMed

This study identified Gram-positive bacteria in three sub-tropical marine fish species; Pseudocaranx dentex (silver trevally), Pagrus auratus (snapper) and Mugil cephalus (sea mullet). It further elucidated the role played by fish habitat, fish body part and ambient storage on the composition of the Gram-positive bacteria. A total of 266 isolates of Gram-positive bacteria were identified by conventional biochemical methods, VITEK, PCR using genus- and species-specific primers and/or 16S rRNA gene sequencing. The isolates were found to fall into 13 genera and 30 species. In fresh fish, Staphylococcus epidermidis and Micrococcus luteus were the most frequent isolates. After ambient storage, S. epidermidis, S. xylosus and Bacillus megaterium were no longer present whereas S. warneri, B. sphaericus, Brevibacillus borstelensis, Enterococcus faecium and Streptococcus uberis increased in frequency. Micrococcus luteus and S. warneri were the most prevalent isolates from P. dentex, while E. faecium and Strep. uberis were the most frequent isolates from P. auratus and M. cephalus. With respect to different parts of the fish body, E. faecium, Strep. uberis and B. sphaericus were the most frequent isolates from the muscles, E. faecium, Strep. uberis from the gills and M. luteus from the gut. This study showed a diversity of Gram-positive bacteria in sub-tropical marine fish; however, their abundance was affected by fish habitat, fish body part and ambient storage. PMID:20110133

Al Bulushi, Ismail M; Poole, Susan E; Barlow, Robert; Deeth, Hilton C; Dykes, Gary A

2010-03-31

94

Surface antigen, SpaA, of erysipelothrix rhusiopathiae binds to Gram-positive bacterial cell surfaces.  

PubMed

In a previous study, we isolated the spaA gene encoding the surface protective antigen A, SpaA, of Erysipelothrix rhusiopathiae, and found that the N-terminal region of SpaA was responsible for protective immunity against erysipelas and that the C-terminal region contained eight repeat units consisting of 20 amino acids comprising the binding domain on the Erysipelothrix cell surface. In this study, using recombinant SpaA proteins, we showed that the repeat region bound to the cell surfaces of various Gram-positive bacterial cells, SpaA was a membrane-associated protein, this association depended on the interaction with choline residues in teichoic acid, and SpaA bound to lipoteichoic acid (LTA) of Bacillus subtilis and Staphylococcus aureus. These results showed that LTA was required for the surface association of SpaA in E. rhusiopathiae and that such an association might be common among Gram-positive bacterial cells. We suggested that an LTA-SpaA complex might have an important role in the E. rhusiopathiae infection process. PMID:10802190

Makino, S I; Yamamoto, K; Asakura, H; Shirahata, T

2000-05-15

95

Acquired inducible antimicrobial resistance in Gram-positive bacteria  

PubMed Central

A major contributor to the emergence of antibiotic resistance in Gram-positive bacterial pathogens is the expansion of acquired, inducible genetic elements. Although acquired, inducible antibiotic resistance is not new, the interest in its molecular basis has been accelerated by the widening distribution and often ‘silent’ spread of the elements responsible, the diagnostic challenges of such resistance and the mounting limitations of available agents to treat Gram-positive infections. Acquired, inducible antibiotic resistance elements belong to the accessory genome of a species and are horizontally acquired by transformation/recombination or through the transfer of mobile DNA elements. The two key, but mechanistically very different, induction mechanisms are: ribosome-sensed induction, characteristic of the macrolide–lincosamide–streptogramin B antibiotics and tetracycline resistance, leading to ribosomal modifications or efflux pump activation; and resistance by cell surface-associated sensing of ?-lactams (e.g., oxacillin), glycopeptides (e.g., vancomycin) and the polypeptide bacitracin, leading to drug inactivation or resistance due to cell wall alterations.

Chancey, Scott T; Zahner, Dorothea; Stephens, David S

2012-01-01

96

Special features of gram-positive bacterial eradication by photosensitizers.  

PubMed

Antibiotic resistance of pathogenic bacteria is a major concern and presents a special challenge for development of alternative antibacterial modalities. One of these alternative approaches is based on using the photodynamic therapy (PDT) for eradicating bacteria. Photosensitizer-induced PDT exhibits unique properties and demonstrates efficient microbe-killing effects. The efficient and irreversible antimicrobial effects of PDT are not dependent on the antibiotic susceptibility of the pathogenic bacteria to antibiotics. Gram-positive bacteria exhibit efficient binding of the photosensitizer to the bacterial barriers, leading to immediate photoinactivation of the bacteria. Photoinactivation of Gram-positive bacteria by various photosensitizers has become a high priority, since these bacteria are responsible for life-threatening infections in humans, especially in the elderly and in compromised hosts in whom they cause hospital-acquired infections. The present review concentrates on the photoinactivation of Staphylococi, Streptococci, Propionibacterium acnes, Deinococcus radiodurans, aerobic spore-forming Bacilli by various photosensitizers and by various methods described in numerous works and patents. PMID:23550546

Nitzan, Yeshayahu; Nisnevitch, Marina

2013-08-01

97

Filamentous Phage Active on the Gram-Positive Bacterium Propionibacterium freudenreichii  

PubMed Central

We present the first description of a single-stranded DNA filamentous phage able to replicate in a gram-positive bacterium. Phage B5 infects Propionibacterium freudenreichii and has a genome consisting of 5,806 bases coding for 10 putative open reading frames. The organization of the genome is very similar to the organization of the genomes of filamentous phages active on gram-negative bacteria. The putative coat protein exhibits homology with the coat proteins of phages PH75 and Pf3 active on Thermus thermophilus and Pseudomonas aeruginosa, respectively. B5 is, therefore, evolutionarily related to the filamentous phages active on gram-negative bacteria.

Chopin, Marie-Christine; Rouault, Annette; Ehrlich, S. Dusko; Gautier, Michel

2002-01-01

98

Gram-positive, catalase-positive cocci from dry cured Iberian ham and their enterotoxigenic potential.  

PubMed Central

Iberian ham is an uncooked, cured meat product ripened under natural uncontrolled conditions for 18 to 24 months. Gram-positive, catalase-positive cocci are the main microbial population in Iberian ham for most of the ripening time. Since some of these organisms are able to produce enterotoxins, adequate characterization and toxicological study are needed. For this, 1,327 gram-positive, catalase-positive cocci, isolated from Iberian hams at different stages and locations, were characterized by physiological and biochemical tests. Selected isolates were further characterized by guanine-cytosine (G+C) content and restriction enzyme analysis of genes coding for 16S rRNA. The toxigenic potential of these organisms was tested with specific DNA gene probes for staphylococcal enterotoxins A, B, C, and D and confirmed by semiquantitative sandwich enzyme immunoassay. The majority of the isolates were identified as Staphylococcus spp. and Micrococcus spp. Non-identified gram-positive, catalase-positive cocci which were moderately halophilic and showed a 42 to 52% G+C content were detected. A great variety of staphylococcal strains were found within the different species at any sampling time. Two strains of Staphylococcus xylosus, one Staphylococcus cohnii strain, and four of the non-identified organisms with 42 to 52% G+C contents hybridized with some of the DNA probes for C and D staphylococcal enterotoxin genes. S. xylosus hybridizing with C-enterotoxin probe reacted with both C and D enterotoxins in the immunological test. In addition, enterotoxin D was confirmed in the nonidentified strains. Some toxigenic organisms were isolated from the final product, posing a health hazard for the consumer.

Rodriguez, M; Nunez, F; Cordoba, J J; Bermudez, E; Asensio, M A

1996-01-01

99

Computational identification of the Spo0A-phosphate regulon that is essential for the cellular differentiation and development in Gram-positive spore-forming bacteria  

PubMed Central

Spo0A-phosphate is essential for the initiation of cellular differentiation and developmental processes in Gram-positive spore-forming bacteria. Here we combined comparative genomics with analyses of microarray expression profiles to identify the Spo0A-phosphate regulon in Bacillus subtilis. The consensus Spo0A-phosphate DNA-binding motif identified from the training set based on different computational algorithms is an 8 bp sequence, TTGTCGAA. The same motif was identified by aligning the upstream regulatory sequences of spo0A-dependent genes obtained from the expression profile of Sad67 (a constitutively active form of Spo0A) and their orthologs. After the transcription units (TUs) having putative Spo0A-phosphate binding sites were obtained, conservation of regulons among the genomes of B.subtilis, Bacillus halodurans and Bacillus anthracis, and expression profiles were employed to identify the most confident predictions. Besides genes already known to be directly under the control of Spo0A-phosphate, 276 novel members (organized in 109 TUs) of the Spo0A-phosphate regulon in B.subtilis are predicted in this study. The sensitivity and specificity of our predictions are estimated based on known sites and combinations of different types of evidence. Further characterization of the novel candidates will provide information towards understanding the role of Spo0A-phosphate in the sporulation process, as well as the entire genetic network governing cellular differentiation and developmental processes in B.subtilis.

Liu, Jiajian; Tan, Kai; Stormo, Gary D.

2003-01-01

100

Coordinate regulation of Gram-positive cell surface components  

PubMed Central

The cell surface of Gram-positive pathogens represents a complex association of glycopolymers that control cell division, homeostasis, immune evasion, tissue invasion, and resistance to antimicrobials. These glycopolymers include the peptidoglycan cell wall, wall-teichoic acids, lipoteichoic acids, and capsular polysaccharide. Disruption of individual factors often results in pleiotropic effects, making it difficult to discern regulation and function. In this review we collate recent work describing these pleiotropic phenotypes, and propose that this is due to coordinated regulation of biosynthesis or modification of these cell surface components. A better understanding of the regulatory networks that control the relative prevalence of each factor on the cell surface or their modulated functions may help facilitate the identification of new targets for antimicrobial therapy.

Hanson, Brett R.; Neely, Melody N.

2012-01-01

101

Nonsporing, anaerobic, gram-positive rods in saliva and the gingival crevice of humans.  

PubMed Central

Quantitative and qualitative examination of anaerobically isolated flora of the gingival crevice and saliva was carried out. It was found that half the organisms were anaerobes and that there were twice as many gram-positive organisms as there were gram-negative ones. Rods were predominant in the gingival crevice (60.5%) and cocci in saliva (69.1%). Of the total organisms, nonsporing, gram-positive anaerobic rods accounted for 24% in the gingival crevice and 9.7% in saliva. These organisms were characterized on the basis of the type of fatty acids produced from glucose and various biochemical reactions. They belonged to the following genera: Actinomyces, Propionibacterium, Arachnia, Lactobacillus, Eubacterium, and Bifidobacterium. Bifidobacteria were present only in saliva. Although members of the other genera were present both in the gingival crevice and saliva, there were considerable differences in the proportion of any particular organism (in relation to the total anaerobic viable count) between the two sites. The result of this study also indicates a greater than previously appreciated level of Propionibacterium and Arachnia in the human mouth.

Sanyal, B; Russell, C

1978-01-01

102

Desulfotomaculum spp. and related gram-positive sulfate-reducing bacteria in deep subsurface environments  

PubMed Central

Gram-positive spore-forming sulfate reducers and particularly members of the genus Desulfotomaculum are commonly found in the subsurface biosphere by culture based and molecular approaches. Due to their metabolic versatility and their ability to persist as endospores. Desulfotomaculum spp. are well-adapted for colonizing environments through a slow sedimentation process. Because of their ability to grow autotrophically (H2/CO2) and produce sulfide or acetate, these microorganisms may play key roles in deep lithoautotrophic microbial communities. Available data about Desulfotomaculum spp. and related species from studies carried out from deep freshwater lakes, marine sediments, oligotrophic and organic rich deep geological settings are discussed in this review.

Aullo, Thomas; Ranchou-Peyruse, Anthony; Ollivier, Bernard; Magot, Michel

2013-01-01

103

Comparative in vitro activity profile of oritavancin against recent gram-positive clinical isolates.  

PubMed

Oritavancin activity was tested against 15,764 gram-positive isolates collected from 246 hospital centers in 25 countries between 2005 and 2008. Organisms were Staphylococcus aureus (n = 9,075), coagulase-negative staphylococci (n = 1,664), Enterococcus faecalis (n = 1,738), Enterococcus faecium (n = 819), Streptococcus pyogenes (n = 959), Streptococcus agalactiae (n = 415), group C, G, and F streptococci (n = 84), and Streptococcus pneumoniae (n = 1,010). Among the evaluated staphylococci, 56.7% were resistant to oxacillin. The vancomycin resistance rate among enterococci was 21.2%. Penicillin-resistant and -intermediate rates were 14.7% and 21.4%, respectively, among S. pneumoniae isolates. Among nonpneumococcal streptococci, 18.5% were nonsusceptible to erythromycin. Oritavancin showed substantial in vitro activity against all organisms tested, regardless of resistance profile. The maximum oritavancin MIC against all staphylococci tested (n = 10,739) was 4 microg/ml; the MIC(90) against S. aureus was 0.12 microg/ml. Against E. faecalis and E. faecium, oritavancin MIC(90)s were 0.06 and 0.12, respectively. Oritavancin was active against glycopeptide-resistant enterococci, including VanA strains (n = 486), with MIC(90)s of 0.25 and 1 microg/ml against VanA E. faecium and E. faecalis, respectively. Oritavancin showed potent activity against streptococci (n = 2,468); MIC(90)s for the different streptococcal species were between 0.008 and 1 microg/ml. These data are consistent with previous studies with respect to resistance rates of gram-positive isolates and demonstrate the spectrum and in vitro activity of oritavancin against a wide variety of contemporary gram-positive pathogens, regardless of resistance to currently used drugs. The data provide a foundation for interpreting oritavancin activity and potential changes in susceptibility over time once oritavancin enters into clinical use. PMID:19738026

Arhin, Francis F; Draghi, Deborah C; Pillar, Chris M; Parr, Thomas R; Moeck, Gregory; Sahm, Daniel F

2009-11-01

104

Comparative In Vitro Activity Profile of Oritavancin against Recent Gram-Positive Clinical Isolates?  

PubMed Central

Oritavancin activity was tested against 15,764 gram-positive isolates collected from 246 hospital centers in 25 countries between 2005 and 2008. Organisms were Staphylococcus aureus (n = 9,075), coagulase-negative staphylococci (n = 1,664), Enterococcus faecalis (n = 1,738), Enterococcus faecium (n = 819), Streptococcus pyogenes (n = 959), Streptococcus agalactiae (n = 415), group C, G, and F streptococci (n = 84), and Streptococcus pneumoniae (n = 1,010). Among the evaluated staphylococci, 56.7% were resistant to oxacillin. The vancomycin resistance rate among enterococci was 21.2%. Penicillin-resistant and -intermediate rates were 14.7% and 21.4%, respectively, among S. pneumoniae isolates. Among nonpneumococcal streptococci, 18.5% were nonsusceptible to erythromycin. Oritavancin showed substantial in vitro activity against all organisms tested, regardless of resistance profile. The maximum oritavancin MIC against all staphylococci tested (n = 10,739) was 4 ?g/ml; the MIC90 against S. aureus was 0.12 ?g/ml. Against E. faecalis and E. faecium, oritavancin MIC90s were 0.06 and 0.12, respectively. Oritavancin was active against glycopeptide-resistant enterococci, including VanA strains (n = 486), with MIC90s of 0.25 and 1 ?g/ml against VanA E. faecium and E. faecalis, respectively. Oritavancin showed potent activity against streptococci (n = 2,468); MIC90s for the different streptococcal species were between 0.008 and 1 ?g/ml. These data are consistent with previous studies with respect to resistance rates of gram-positive isolates and demonstrate the spectrum and in vitro activity of oritavancin against a wide variety of contemporary gram-positive pathogens, regardless of resistance to currently used drugs. The data provide a foundation for interpreting oritavancin activity and potential changes in susceptibility over time once oritavancin enters into clinical use.

Arhin, Francis F.; Draghi, Deborah C.; Pillar, Chris M.; Parr, Thomas R.; Moeck, Gregory; Sahm, Daniel F.

2009-01-01

105

Synthesis and in vitro evaluation of substituted phenyl-piperazinyl-phenyl oxazolidinones against Gram-positive bacteria.  

PubMed

With the incidence of linezolid-resistant Enterococcus faecalis, E. faecium and Staphylococcus aureus, modification of linezolid at the 5- and/or 3-positions led to the development of a series of 3-(methoxyl-phenyl)-piperazinyl-phenyl oxazolidinone analogues. These compounds were tested in vitro against six gram-positive standard organisms (S. aureus, S. epidermidis, S. pneumoniae, S. albus, Streptococcus enteridis and S. nonhemolyticus). 5-acetylaminomethyl oxazolidinones bearing fluorine at 3'-position of phenyl ring showed activities against several gram-positive bacteria (MIC: 3.13-6.25 mug/mL). The position of methoxyl group on the phenyl ring of piperazine group affected antibacterial spectrum. 3-(4'- (para-methoxyl-phenyl)-piperazinyl)-(3'-fluoro)-phenyl-5-acetylaminomethyl oxazolidinone was found active against 5 gram-positive organisms except S. nonhemolyticus, whereas 3-(4'-(ortho-methoxyl-phenyl)-piperazinyl)-(3'-fluoro)-phenyl-5-acetylaminomethyl oxazolidinone was found active only against 2 gram-positive organisms, namely S. albus, S. enteridis. PMID:17461974

Liu, Jidong; He, Baoyuan; Yu, Aizhen; Zhou, Weicheng

2007-04-01

106

Effects of Ultraviolet Radiation on the Gram-positive marine bacterium Microbacterium maritypicum.  

PubMed

Although extensive information is available on the effect ultraviolet (UV) radiation has on Gram-negative marine bacteria, there is a scarcity of data concerning UV radiation and Gram-positive marine bacteria. The focus of this paper is on Microbacterium maritypicum, with the Gram-negative Vibrio natriegens being used as a standard of comparison. M. maritypicum exhibited growth over a NaCl range of 0-1000 mM: , with optimum growth occurring between 0 and 400 mM: NaCl. In contrast, V. natriegens grew over a NaCl span of 250-1000 mM: , with best growth being observed between 250 and 600 mM: NaCl. UV radiation experiments were done using the medium with 250 mM: NaCl. For solar (UV-A and B) radiation and log-phase cells, M. maritypicum was determined to be three times more resistant than V. natriegens. For germicidal (UV-C) radiation, the pattern of resistance of the log-phase cells to the lethal effects of the radiation was even more pronounced, with the Gram-positive bacterium being more than 12 to 13 times more resistant. Similar data to the solar and germicidal log-phase UV kill curves were obtained for stationary-phase cells of both organisms. Photoreactivation was observed for both types of cells exposed to UV-C but none for cells treated with UV-A and B. When log phase cells of M.maritypicum were grown at 0.0 and 0.6 M: NaCl and exposed to UV-C radiation, no difference in survivorship patterns was noted from that of 0.25 M: NaCl grown cells. Although this study has only focused on two marine bacteria, our results indicate that the Gram-positive M. maritypicum could have a built-in advantage for survival in some marine ecosystems. PMID:17551790

Williams, Patrick D; Eichstadt, Shaundra L; Kokjohn, Tyler A; Martin, Eugene L

2007-07-01

107

SubtiWiki-a database for the model organism Bacillus subtilis that links pathway, interaction and expression information.  

PubMed

Genome annotation and access to information from large-scale experimental approaches at the genome level are essential to improve our understanding of living cells and organisms. This is even more the case for model organisms that are the basis to study pathogens and technologically important species. We have generated SubtiWiki, a database for the Gram-positive model bacterium Bacillus subtilis (http://subtiwiki.uni-goettingen.de/). In addition to the established companion modules of SubtiWiki, SubtiPathways and SubtInteract, we have now created SubtiExpress, a third module, to visualize genome scale transcription data that are of unprecedented quality and density. Today, SubtiWiki is one of the most complete collections of knowledge on a living organism in one single resource. PMID:24178028

Michna, Raphael H; Commichau, Fabian M; Tödter, Dominik; Zschiedrich, Christopher P; Stülke, Jörg

2014-01-01

108

SubtiWiki-a database for the model organism Bacillus subtilis that links pathway, interaction and expression information  

PubMed Central

Genome annotation and access to information from large-scale experimental approaches at the genome level are essential to improve our understanding of living cells and organisms. This is even more the case for model organisms that are the basis to study pathogens and technologically important species. We have generated SubtiWiki, a database for the Gram-positive model bacterium Bacillus subtilis (http://subtiwiki.uni-goettingen.de/). In addition to the established companion modules of SubtiWiki, SubtiPathways and SubtInteract, we have now created SubtiExpress, a third module, to visualize genome scale transcription data that are of unprecedented quality and density. Today, SubtiWiki is one of the most complete collections of knowledge on a living organism in one single resource.

Michna, Raphael H.; Commichau, Fabian M.; Todter, Dominik; Zschiedrich, Christopher P.; Stulke, Jorg

2014-01-01

109

Biochemical characterization of Gram-positive and Gram-negative plant-associated bacteria with micro-Raman spectroscopy.  

PubMed

Raman spectra of Gram-positive and Gram-negative plant bacteria have been measured with micro-Raman spectrometers equipped with 785 and 514.5 nm lasers. The Gram-positive bacteria Microbacterium testaceum, Paenibacillus validus, and Clavibacter michiganensis subsp. michiganensis have strong carotenoid bands in the regions 1155-1157 cm(-1) and 1516-1522 cm(-1) that differentiate them from other tested Gram-negative bacteria. In the Raman spectrum of Gram-positive bacteria Bacillus megaterium excited with 785 nm laser, the Raman bands at 1157 and 1521 cm(-1) are weak in intensity compared to other Gram-positive bacteria, and these bands did not show significant resonance Raman enhancement in the spectrum recorded with 514.5 nm laser excitation. The Gram-positive bacteria could be separated from each other based on the bands associated with the in-phase C=C (v(1)) vibrations of the polyene chain of carotenoids. None of the Gram-negative bacteria tested had carotenoid bands. The bacteria in the genus Xanthomonas have a carotenoid-like pigment, xanthomonadin, identified in Xanthomonas axonopodis pv. dieffenbachiae, and it is a unique Raman marker for the bacteria. The representative bands for xanthomonadin were the C-C stretching (v(2)) vibrations of the polyene chain at 1135-1136 cm(-1) and the in-phase C=C (v(1)) vibrations of the polyene chain at 1529-1531 cm(-1), which were distinct from the carotenoid bands of other tested bacteria. The tyrosine peak in the region 1170-1175 cm(-1) was the only other marker present in Gram-negative bacteria that was absent in all tested Gram-positives. A strong-intensity exopolysaccharide-associated marker at 1551 cm(-1) is a distinguishable feature of Enterobacter cloacae. The Gram-negative Agrobacterium rhizogenes and Ralstonia solanacearum were differentiated from each other and other tested bacteria on the basis of presence or absence and relative intensities of peaks. The principal components analysis (PCA) of the spectra excited with 785 nm laser differentiated the various strains of bacteria based on the unique pigments these bacteria do or do not possess. Raman spectroscopy of diverse plant bacteria that are pathogenic and non-pathogenic to plants, and isolated from plants and soil, indicates the possibilities of using the method in understanding plant-bacterial interactions at the cellular level. PMID:20412629

Paret, Mathews L; Sharma, Shiv K; Green, Lisa M; Alvarez, Anne M

2010-04-01

110

Sulrfobacillus disuljidooxidans sp. nov., a New Acidophilic, Disulfide-Oxidizing, Gram-Positive, Spore-Forming Bacterium  

Microsoft Academic Search

An acidophilic, disulfide-oxidizing, mesophilic, aerobic bacterium was isolated from wastewater sludge. The new organism is a gram-positive sporulated rod. It can use elemental sulfur and pyrite as sole energy sources and grows on organic substrates such as glutamate and glucose. It also grows on the following organic sulfur substrates: oxidized and reduced glutathione, cysteine, cystine, and dithio(bis) benzothiazole and clearly

S. DUFRESNE; J. BOUSQUET; M. BOISSINOT; R. GUAY; Universitd Laval

111

Characterization of a pyridine-degrading branched Gram-positive bacterium isolated from the anoxic zone of an oil shale column  

Microsoft Academic Search

From the anoxic zone of an oil shale leachate column three pyridine-degrading bacterial strains were isolated. Two strains were Gram-negative facultative anaerobic rods and one strain was a branched Gram-positive bacterium. The branched Gram-positive strain had the best pyridine-degrading ability. This organism was aerobic, non-motile, catalase positive, oxidase negative, and had no flagellum. The G+C content of the DNA was

Sung-Taik Lee; Seung-Bong Lee; Yong-Ha Park

1991-01-01

112

Novel Bacterial Lipoprotein Structures Conserved in Low-GC Content Gram-positive Bacteria Are Recognized by Toll-like Receptor 2*  

PubMed Central

Bacterial lipoproteins/lipopeptides inducing host innate immune responses are sensed by mammalian Toll-like receptor 2 (TLR2). These bacterial lipoproteins are structurally divided into two groups, diacylated or triacylated lipoproteins, by the absence or presence of an amide-linked fatty acid. The presence of diacylated lipoproteins has been predicted in low-GC content Gram-positive bacteria and mycoplasmas based on the absence of one modification enzyme in their genomes; however, we recently determined triacylated structures in low-GC Gram-positive Staphylococcus aureus, raising questions about the actual lipoprotein structure in other low-GC content Gram-positive bacteria. Here, through intensive MS analyses, we identified a novel and unique bacterial lipoprotein structure containing an N-acyl-S-monoacyl-glyceryl-cysteine (named the lyso structure) from low-GC Gram-positive Enterococcus faecalis, Bacillus cereus, Streptococcus sanguinis, and Lactobacillus bulgaricus. Two of the purified native lyso-form lipoproteins induced proinflammatory cytokine production from mice macrophages in a TLR2-dependent and TLR1-independent manner but with a different dependence on TLR6. Additionally, two other new lipoprotein structures were identified. One is the “N-acetyl” lipoprotein structure containing N-acetyl-S-diacyl-glyceryl-cysteine, which was found in five Gram-positive bacteria, including Bacillus subtilis. The N-acetyl lipoproteins induced the proinflammatory cytokines through the TLR2/6 heterodimer. The other was identified in a mycoplasma strain and is an unusual diacyl lipoprotein structure containing two amino acids before the lipid-modified cysteine residue. Taken together, our results suggest the existence of novel TLR2-stimulating lyso and N-acetyl forms of lipoproteins that are conserved in low-GC content Gram-positive bacteria and provide clear evidence for the presence of yet to be identified key enzymes involved in the bacterial lipoprotein biosynthesis.

Kurokawa, Kenji; Ryu, Kyoung-Hwa; Ichikawa, Rie; Masuda, Akiko; Kim, Min-Su; Lee, Hanna; Chae, Jun-Ho; Shimizu, Takashi; Saitoh, Tatsuya; Kuwano, Koichi; Akira, Shizuo; Dohmae, Naoshi; Nakayama, Hiroshi; Lee, Bok Luel

2012-01-01

113

Anaerobic naphthalene degradation by Gram-positive, iron-reducing bacteria.  

PubMed

An anaerobic naphthalene-degrading culture (N49) was enriched with ferric iron as electron acceptor. A closed electron balance indicated the total oxidation of naphthalene to CO(2). In all growing cultures, the concentration of the presumed central metabolite of naphthalene degradation, 2-naphthoic acid, increased concomitantly with growth. The first metabolite of anaerobic methylnaphthalene degradation, naphthyl-2-methyl-succinic acid, was not identified in culture supernatants, which does not support a methylation to methylnaphthalene as the initial activation reaction of naphthalene, but rather a carboxylation, as proposed for other naphthalene-degrading cultures. Substrate utilization tests revealed that the culture was able to grow on 1-methyl-naphthalene, 2-methyl-naphthalene, 1-naphthoic acid or 2-naphthoic acid, whereas it did not grow on 1-naphthol, 2-naphthol, anthracene, phenanthrene, indane and indene. Terminal restriction fragment length polymorphism and 16S rRNA gene sequence analyses revealed that the microbial community of the culture was dominated by one bacterial microorganism, which was closely related (99% 16S sequence similarity) to the major organism in the iron-reducing, benzene-degrading enrichment culture BF [ISME J (2007) 1: 643; Int J Syst Evol Microbiol (2010) 60: 686]. The phylogenetic classification supports a new candidate species and genus of Gram-positive spore-forming iron-reducers that can degrade non-substituted aromatic hydrocarbons. It furthermore indicates that Gram-positive microorganisms might also play an important role in anaerobic polycyclic aromatic hydrocarbon-degradation. PMID:22066721

Kleemann, Rita; Meckenstock, Rainer U

2011-12-01

114

Thermophilic Gram-Positive Biocatalysts for Biomass Conversion to Ethanol  

SciTech Connect

Production of energy from renewable sources is receiving increased attention due to the finite nature of fossil fuels and the environmental impact associated with the continued large scale use of fossil energy sources. Biomass, a CO2-neutral abundant resource, is an attractive alternate source of energy. Biomass-derived sugars, such as glucose, xylose, and other minor sugars, can be readily fermented to fuel ethanol and commodity chemicals. Extracellular cellulases produced by fungi are commercially developed for depolymerization of cellulose in biomass to glucose for fermentation by appropriate biocatalysts in a simultaneous saccharification and fermentation (SSF) process. Due to the differences in the optimum conditions for the activity of the fungal cellulases and the growth and fermentation characteristics of the current industrial biocatalysts, SSF of cellulose is envisioned at conditions that are not optimal for the fungal cellulase activity leading to higher than required cost of cellulase in SSF. We have isolated bacterial biocatalysts whose growth and fermentation requirements match the optimum conditions for commercial fungal cellulase activity (pH 5.0 and 50 deg. C). These isolates fermented both glucose and xylose, major components of cellulose and hemicellulose, respectively, to L(+)-lactic acid. Xylose was metabolized through the pentose-phosphate pathway by these organisms as evidenced by the fermentation profile and analysis of the fermentation products of 13C1-xylose by NMR. As expected for the metabolism of xylose by the pentose-phosphate pathway, 13C-lactate accounted for more than 90% of the total 13C-labeled products. All three strains fermented crystalline cellulose to lactic acid with the addition of fungal cellulase (Spezyme CE) (SSF) at an optimum of about 10 FPU/g cellulose. These isolates also fermented cellulose and sugar cane bagasse hemicellulose acid hydrolysate simultaneously. Based on fatty acid profile and 16S rRNA sequence, these isolates cluster with Bacillus coagulans although B. coagulans type strain, ATCC 7050, failed to utilize xylose as a carbon source. For successful production of ethanol from pyruvate, both pyruvate decarboxylase (PDC) and alcohol dehydrogenase (AHD) need to be produced at optimal levels in these biocatalysts. A plasmid containing the S. ventriculi pdc gene and the adh gene from geobacillus stearothermophilus was constructed using plasmid pWH1520 that was successfully used for expression of pdc in B. megaterium. The resulting portable ethanol (PET) plasmid, pJAM423, was transformed into B. megaterium. After xylose induction, a significant fraction of cell cytoplasm was composed of the S. ventriculi PDC and G. stearothermophilus ADH proteins. In preliminary experiments, the amount of ethanol produced by b. megaterium with plasmid pJAM423 was about twice (20 mM) of the bacterium without the plasmid. These results show that the PET operon is functional in B. megaterium but high level ethanol production needs further genetic and metabolic engineering. A genetic transfer system for the second generation biocatalysts needs to be developed for transferring the plasmid pJAM423 and its derivatives for engineering these organisms for ethanol production from biomass derived sugars and cellulose to ethanol. One of the new biocatalysts, strain P4-102B was found to be transformable with plasmids and the method for introducing plasmid pJAM423 into this strain and expression of the encoded DNA is being optimized. These new second generation biocatalysts have the potential to reduce the cost of SSF by minimizing the amount of fungal cellulases, a significant cost component in the use of biomass as a renewable resource for production of fuels and chemicals.

Shanmugam, K.T.; Ingram, L.O.; Maupin-Furlow, J.A.; Preston, J.F.; Aldrich, H.C.

2003-12-01

115

Rhamnolipid-biosurfactant permeabilizing effects on gram-positive and gram-negative bacterial strains.  

PubMed

The potential of biosurfactant PS to permeabilize bacterial cells of Pseudomonas aeruginosa, Escherichia coli, and Bacillus subtilis on growing (in vivo) and resting (in vitro) cells was studied. Biosurfactant was shown to have a neutral or detrimental effect on the growth of Gram-positive strains, and this was dependent on the surfactant concentration. The growth of Gram-negative strains was not influenced by the presence of biosurfactant in the media. Cell permeabilization with biosurfactant PS was shown to be more effective with B. subtilis resting cells than with Pseudomonas aeruginosa. Scanning-electron microscopy observations showed that the biosurfactant PS did not exert a disruptive action on resting cells such that it was detrimental to the effect on growing cells of B. subtilis. Low critical micelle concentrations, tender action on nongrowing cells, and neutral effects on the growth of microbial strains at low surfactant concentrations make biosurfactant PS a potential candidate for application in different industrial fields, in environmental bioremediation, and in biomedicine. PMID:18330632

Sotirova, A V; Spasova, D I; Galabova, D N; Karpenko, E; Shulga, A

2008-06-01

116

Antibacterial activity of oregano (Origanum vulgare Linn.) against gram positive bacteria.  

PubMed

The present investigation is focused on antibacterial potential of infusion, decoction and essential oil of oregano (Origanum vulgare) against 111 Gram-positive bacterial isolates belonging to 23 different species related to 3 genera. Infusion and essential oil exhibited antibacterial activity against Staphylococcus saprophyticus, S. aureus, Micrococcus roseus, M. kristinae, M. nishinomiyaensis, M. lylae, M. luteus, M. sedentarius, M. varians, Bacillus megaterium, B. thuringiensis, B. alvei, B. circulans, B. brevis, B. coagulans, B. pumilus, B. laterosporus, B. polymyxa, B. macerans, B. subtilis, B. firmus, B. cereus and B. lichiniformis. The infusion exhibited maximum activity against B. laterosporus (17.5 mm mean zone of inhibition+/-1.5 Standard deviation) followed by B. polymyxa (17.0 mm+/-2.0 SD) and essential oil of oregano exhibited maximum activity against S. saprophyticus (16.8 mm+/-1.8 SD) followed by B. circulans (14.5 mm+/-0.5 SD). While all these tested isolates were found resistant to decoction of oregano. PMID:19783523

Saeed, Sabahat; Tariq, Perween

2009-10-01

117

Dry and moist heat sterilisation cannot inactivate pyrogenicity of Gram positive microorganisms.  

PubMed

In the monocytic cell line Mono Mac 6 pyrogens induce interleukin-6 secretion dose dependently. The aim of this study is to examine the interleukin-6 inducing capacity of Gram positive Staphylococcus aureus and Bacillus subtilis endospores after moist and dry heat sterilisation. Moist heat sterilisation of B. subtilis endospores for 15 min at 121 degrees C and 134 degrees C can only reduce the interleukin-6 inducing capacity to 57% and 63%, respectively, compared to untreated. Moist heat sterilisation of S. aureus for 60 min at 121 degrees C and 134 degrees C does not reduce the interleukin-6 inducing capacity of S. aureus. On the contrary moist heat sterilisation at 134 degrees C for 10, 20 and 40 min significantly increases the interleukin-6 inducing capacity compared to untreated S. aureus. This is confirmed in the rabbit pyrogen test. Dry heat sterilisation of B. subtilis endospores at 220 degrees C for 45 min reduces the interleukin-6 inducing capacity to 2% compared to untreated endospores. Dry heat treatment of S. aureus at 220 degrees C for 30 min only reduces the activity to 55%. However, after 250 degrees C for 30 min or 220 degrees C for 6h there is no detectable activity of S. aureus. In conclusion, neither the interleukin-6 inducing activity nor the pyrogenicity of S. aureus and endospores of B. subtilis can be inactivated by the heat sterilisation procedures described by the European Pharmacopoeia. PMID:16125917

Moesby, Lise; Hansen, Erik W; Christensen, Jens D; Høyer, Catrine H; Juhl, Gitte L; Olsen, Helle B

2005-11-01

118

[Isolation of Gram-positive bacteria from raw milk with antimicrobial residues].  

PubMed

Two hundred samples of raw milk were collected at the receiving plants located in three areas of high milk production in Zulia state, Venezuela. The CTT test and trial disk were used in order to detect the presence of antimicrobials. The positive samples were inoculated in tripticase soy broth, human blood agar and manitol salt agar in order to isolate Gram-positive bacteria. The identification of species was performed through biochemical tests. It was found that 45 samples (22.5%) of analyzed milk contained antimicrobials, and bacterial growth was obtained in 35 of them. 100 strains were isolated namely: 44 Staphylococcus, 19 Streptococcus, 17 Enterococcus, 9 Bacillus, 4 Micrococcus, 4 Corynebacterium and 3 Lactococcus. The most frequently isolated specie was S. aureus, the main producing agent of bovine mastitis in Zulia state, a microorganism frequently associated in the country to food-borne intoxications, associated to cheese processed from raw milk. It is recommended to apply control programs for the use of antibiotics. PMID:12214550

Faría Reyes, José; García Urdaneta, Aleida; Izquierdo Corser, Pedro; Allara Cagnasso, María; Valero Leal, Kutchynskaya

2002-03-01

119

Cultivation of aerobic chemoorganotrophic proteobacteria and gram-positive bacteria from a hot spring microbial mat.  

PubMed

The diversity of aerobic chemoorganotrophic bacteria inhabiting the Octopus Spring cyanobacterial mat community (Yellowstone National Park) was examined by using serial-dilution enrichment culture and a variety of enrichment conditions to cultivate the numerically significant microbial populations. The most abundant bacterial populations cultivated from dilutions to extinction were obtained from enrichment flasks which contained 9.0 x 10(2) primary producer (Synechococcus spp.) cells in the inoculum. Two isolates exhibited 16S rRNA nucleotide sequences typical of beta-proteobacteria. One of these isolates contained a 16S rRNA sequence identical to a sequence type previously observed in the mat by molecular retrieval techniques. Both are distantly related to a new sequence directly retrieved from the mat and contributed by a beta-proteobacterial community member. Phenotypically diverse gram-positive isolates genetically similar to Bacillus flavothermus were obtained from a variety of dilutions and enrichment types. These isolates exhibited identical 16S rRNA nucleotide sequences through a variable region of the molecule. Of the three unique sequences observed, only one had been previously retrieved from the mat, illustrating both the inability of the cultivation methods to describe the composition of a microbial community and the limitations of the ability of molecular retrieval techniques to describe populations which may be less abundant in microbial communities. PMID:8899976

Nold, S C; Kopczynski, E D; Ward, D M

1996-11-01

120

Antioxidant activity via DPPH, gram-positive and gram-negative antimicrobial potential in edible mushrooms.  

PubMed

Edible mushrooms (EMs) are nutritionally rich source of proteins and essential amino acids. In the present study, the antioxidant activity via 1,1-diphenyl-2-picrylhydrazyl (DPPH) and antimicrobial potential in EMs (Pleurotus ostreatus, Morchella esculenta, P. ostreatus (Black), P. ostreatus (Yellow) and Pleurotus sajor-caju) were investigated. The DPPH radical scavenging activity revealed that the significantly higher activity (66.47%) was observed in Morchella esculenta at a maximum concentration. Similarly, the dose-dependent concentrations (200, 400, 600, 800 and 1000 µg) were also used for other four EMs. Pleurotus ostreatus exhibited 36.13% activity, P. ostreatus (Black (B)) exhibited 30.64%, P. ostreatus (Yellow (Y)) exhibited 40.75% and Pleurotus sajor-caju exhibited 47.39% activity at higher concentrations. Furthermore, the antimicrobial potential were investigated for its toxicity against gram-negative bacterial strains (Escherichia coli, Pseudomonas aeroginosa, Salmonella typhi, Klebsiella pneumonia, Erwinia carotovora and Agrobacterium tumifaciens), gram-positive bacterial strains (Bacillus subtilis, Bacillus atrophaeus and Staphylococcus aureus) and a fungal strain (Candida albicans) in comparison with standard antibiotics. Antimicrobial screening revealed that the ethanol extract of P. ostreatus was active against all microorganism tested except E. coli. Maximum zone of inhibition (13 mm) was observed against fungus and A. tumifaciens. P. sajor-caju showed best activities (12.5 mm) against B. subtilis, B. atrophaeus and K. pneumonia. P. ostreatus (Y) showed best activities against P. aeroginosa (21.83 mm), B. atrophaeus (20 mm) and C. albicans (21 mm). P. ostreatus (B) exhibited best activities against C. albicans (16 mm) and slightly lower activities against all other microbes except S. typhi. M. esculenta possess maximum activities in terms of inhibition zone against all microorganisms tested except S. typhi. PMID:23095488

Ahmad, Nisar; Mahmood, Fazal; Akbar Khalil, Shahid; Zamir, Roshan; Fazal, Hina; Abbasi, Bilal Haider

2012-10-24

121

Aspects of eukaryotic-like signaling in Gram-positive cocci: a focus on virulence  

PubMed Central

Living organisms adapt to the dynamic external environment for their survival. Environmental adaptation in prokaryotes is thought to be primarily accomplished by signaling events mediated by two-component systems, consisting of histidine kinases and response regulators. However, eukaryotic-like serine/threonine kinases (STKs) have recently been described to regulate growth, antibiotic resistance and virulence of pathogenic bacteria. This article summarizes the role of STKs and their cognate phosphatases (STPs) in Gram-positive cocci that cause invasive infections in humans. Given that a large number of inhibitors to eukaryotic STKs are approved for use in humans, understanding how serine/threonine phosphorylation regulates virulence and antibiotic resistance will be beneficial for the development of novel therapeutic strategies against bacterial infections.

Burnside, Kellie; Rajagopal, Lakshmi

2011-01-01

122

Early appropriate therapy of Gram-positive bloodstream infections: the conservative use of new drugs.  

PubMed

Among the Gram-positive organisms, meticillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecium represent the biggest therapeutic hurdles. The evolution of MRSA exemplifies the genetic adaptation of an organism into a first-class multidrug-resistant pathogen. Glycopeptides such as vancomycin have been the treatment of choice for MRSA, but poor outcomes have frequently been reported, particularly among isolates with higher minimum inhibitory concentrations (MICs) within the susceptible range (< or =2 mg/L). Further more, vancomycin's limitations as an antibacterial agent include slow bactericidal activity and relatively poor tissue penetration. Inadequate dosing may, however, contribute to vancomycin's poor performance; the standard recommendation for trough concentrations of 5-10 mg/L is inadequate for serious infections such as bacteraemia and endocarditis. Trough levels of 15-20 mg/L are probably necessary; however they are often associated with increased nephrotoxicity. Despite the recent dramatic reduction in antibiotic research by pharmaceutical companies, a few compounds have been developed to treat Gram-positive infections. Quinupristin-dalfopristin, although shown to have in vitro activity against MRSA, is not approved by the US Food and Drug Administration for the treatment of MRSA, and cannot be recommended for the treatment of S. aureus bacteraemia, except under exceptional circumstances. Although linezolid and tigecycline may be useful in specific situations, they cannot be routinely recommended for the treatment of MRSA bacteraemia because of safety concerns and very limited available clinical data. Daptomycin has recently been proven to be effective and well tolerated for meticillin-sensitive S. aureus and MRSA bacteraemia, including right-sided endocarditis. PMID:19931814

Grossi, Paolo A

2009-01-01

123

Weissella confusa: an unexpected cause of vancomycin-resistant gram-positive bacteremia in immunocompromised hosts.  

PubMed

We report the first case of Weissella confusa bacteremia in an allogeneic hematopoietic stem cell transplant patient. After engraftment and discharge, the patient returned with fever and graft failure and was started on an empiric regimen of aztreonam and vancomycin. A blood culture grew an alpha-hemolytic, gram-positive coccus forming pairs and chains, originally thought to be a viridans Streptococcus and a skin contaminant. The isolation of the organism from multiple blood cultures, and the presence of vancomycin resistance prompted identification and additional susceptibility testing. The RapID(™) Str panel, which has W. confusa in its database, provided multiple incorrect identifications. The MicroScan WalkAway 96 SI, using PC-20 or -29 panels, also did not identify this bacterium, because it is not in their database. The organism was identified as W. confusa by 16S rDNA sequencing. Antibiotic susceptibility determination by Etest revealed vancomycin resistance and daptomycin susceptibility. Therapy was changed to daptomycin, and the infection resolved. Additionally, W. confusa sepsis, with multiple positive blood cultures, developed in a patient in the burn unit at our medical center. The patient's blood cultures remained positive until vancomycin was discontinued and daptomycin therapy initiated. Infections with vancomycin-resistant, gram-positive cocci are emerging among immuno compromised hosts. Under appropriate circumstances, clinicians need to request that the laboratory perform susceptibility testing and accurate identification, by nucleic acid sequencing if necessary. Sequencing of 16S rDNA is an important tool in the accurate identification of unusual pathogens. PMID:21156010

Salimnia, H; Alangaden, G J; Bharadwaj, R; Painter, T M; Chandrasekar, P H; Fairfax, M R

2011-06-01

124

Identification and characterization of cell wall-cell division gene clusters in pathogenic gram-positive cocci.  

PubMed Central

Clusters of peptidoglycan biosynthesis and cell division genes (DCW genes) were identified and sequenced in two gram-positive cocci, Staphylococcus aureus and Enterococcus faecalis. The results indicated some similarities in organization compared with previously reported bacterial DCW gene clusters, including the presence of penicillin-binding proteins at the left ends and ftsA and ftsZ cell division genes at the right ends of the clusters. However, there were also some important differences, including the absence of several genes, the comparative sizes of the div1B and ftsQ genes, and a wide range of amino acid sequence similarities when the genes of the gram-positive cocci were translated and compared to bacterial homologs.

Pucci, M J; Thanassi, J A; Discotto, L F; Kessler, R E; Dougherty, T J

1997-01-01

125

Complete Genome Sequence of Bacillus bombysepticus, a Pathogen Leading to Bombyx mori Black Chest Septicemia.  

PubMed

Bacillus bombysepticus is a Gram-positive spore-forming bacterium. Here, we announce the first complete genome sequence of this organism isolated from the cadavers of silkworm larvae that had been sick. The genome contains a single circular chromosome and a circular plasmid. Analyses of the B. bombysepticus genome will provide insights into its pathomechanisms and biology. PMID:24831136

Cheng, Tingcai; Lin, Ping; Jin, Shengkai; Wu, Yuqian; Fu, Bohua; Long, Renwen; Liu, Duolian; Guo, Youbing; Peng, Li; Xia, Qingyou

2014-01-01

126

Complete Genome Sequence of Bacillus bombysepticus, a Pathogen Leading to Bombyx mori Black Chest Septicemia  

PubMed Central

Bacillus bombysepticus is a Gram-positive spore-forming bacterium. Here, we announce the first complete genome sequence of this organism isolated from the cadavers of silkworm larvae that had been sick. The genome contains a single circular chromosome and a circular plasmid. Analyses of the B. bombysepticus genome will provide insights into its pathomechanisms and biology.

Cheng, Tingcai; Lin, Ping; Jin, Shengkai; Wu, Yuqian; Fu, Bohua; Long, Renwen; Liu, Duolian; Guo, Youbing; Peng, Li

2014-01-01

127

Antimicrobial activity of metal oxide nanoparticles against Gram-positive and Gram-negative bacteria: a comparative study  

PubMed Central

Background Nanomaterials have unique properties compared to their bulk counterparts. For this reason, nanotechnology has attracted a great deal of attention from the scientific community. Metal oxide nanomaterials like ZnO and CuO have been used industrially for several purposes, including cosmetics, paints, plastics, and textiles. A common feature that these nanoparticles exhibit is their antimicrobial behavior against pathogenic bacteria. In this report, we demonstrate the antimicrobial activity of ZnO, CuO, and Fe2O3 nanoparticles against Gram-positive and Gram-negative bacteria. Methods and results Nanosized particles of three metal oxides (ZnO, CuO, and Fe2O3) were synthesized by a sol–gel combustion route and characterized by X-ray diffraction, Fourier-transform infrared spectroscopy, and transmission electron microscopy techniques. X-ray diffraction results confirmed the single-phase formation of all three nanomaterials. The particle sizes were observed to be 18, 22, and 28 nm for ZnO, CuO, and Fe2O3, respectively. We used these nanomaterials to evaluate their antibacterial activity against both Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive (Staphylococcus aureus and Bacillus subtilis) bacteria. Conclusion Among the three metal oxide nanomaterials, ZnO showed greatest antimicrobial activity against both Gram-positive and Gram-negative bacteria used in this study. It was observed that ZnO nanoparticles have excellent bactericidal potential, while Fe2O3 nanoparticles exhibited the least bactericidal activity. The order of antibacterial activity was demonstrated to be the following: ZnO > CuO > Fe2O3.

Azam, Ameer; Ahmed, Arham S; Oves, Mohammad; Khan, Mohammad S; Habib, Sami S; Memic, Adnan

2012-01-01

128

In vitro pyrogenicity of Gram-positive bacteria--validation of the kit using fresh human whole blood.  

PubMed

The two conventional tests to detect pyrogen contaminants in injectable pharmaceutical drugs are the Rabbit Model and the Limulus amoebocyte lysate (LAL) test. To replace these models, a new system on human whole blood is developed, using the release of Interleukin 1 beta (IL1beta) after cell stimulation with gram-positive and gram negative pyrogens. The purpose of this study was to validate the ENDOSAFE-IPT kit using the quantitative ELISA enzyme immunoassay. The assay is divided into two parts: blood cell stimulation with Lipopolysaccharides (LPS) and Lipoteichoic acid (LTA) and quantitation of IL1beta using the ELISA method. In each assay, blood from a particular donor were stimulated with the Endotoxin Standard, and with a sample of a commercial antibiotic preparation (Clavulanic acid/Ticarcillin) spiked with the Endotoxin Standard. LTA from Bacillus subtilis and a sample of diphtheria toxoid were also used. At least, six assays were tested. A polynomial regression of the Endotoxin Standard series showed a correlation coefficient greater than 0.99. The spiked antibiotic sample recoveries were 50-121%. The LTA quantitation limit was 0.1 microg/ml and the range of detection of pyrogens from Gram positive diphtheria toxoid was 0.77 to 2.5 EEU/ml. The IL1beta production varied markedly between donors. However the coefficient of variation was less than 20 % intra-assay. In conclusion, the ENDOSAFE-IPT kit can be used for the quantitative and qualitative detection of pyrogens from Gram negative and Gram positive bacteria. PMID:17694641

François, C; Neveu, J; Sauvaire, D; Bonnet, P A; Tissier, M H

2006-08-01

129

Caenorhabditis elegans immune conditioning with the probiotic bacterium Lactobacillus acidophilus strain NCFM enhances gram-positive immune responses.  

PubMed

Although the immune response of Caenorhabditis elegans to microbial infections is well established, very little is known about the effects of health-promoting probiotic bacteria on evolutionarily conserved C. elegans host responses. We found that the probiotic Gram-positive bacterium Lactobacillus acidophilus NCFM is not harmful to C. elegans and that L. acidophilus NCFM is unable to colonize the C. elegans intestine. Conditioning with L. acidophilus NCFM significantly decreased the burden of a subsequent Enterococcus faecalis infection in the nematode intestine and prolonged the survival of nematodes exposed to pathogenic strains of E. faecalis and Staphylococcus aureus, including multidrug-resistant (MDR) isolates. Preexposure of nematodes to Bacillus subtilis did not provide any beneficial effects. Importantly, L. acidophilus NCFM activates key immune signaling pathways involved in C. elegans defenses against Gram-positive bacteria, including the p38 mitogen-activated protein kinase pathway (via TIR-1 and PMK-1) and the ?-catenin signaling pathway (via BAR-1). Interestingly, conditioning with L. acidophilus NCFM had a minimal effect on Gram-negative infection with Pseudomonas aeruginosa or Salmonella enterica serovar Typhimurium and had no or a negative effect on defense genes associated with Gram-negative pathogens or general stress. In conclusion, we describe a new system for the study of probiotic immune agents and our findings demonstrate that probiotic conditioning with L. acidophilus NCFM modulates specific C. elegans immunity traits. PMID:22585961

Kim, Younghoon; Mylonakis, Eleftherios

2012-07-01

130

Antimicrobial Activities of Leaf Extracts of Guava (Psidium guajava L.) on Two Gram-Negative and Gram-Positive Bacteria  

PubMed Central

Aim. To determine the antimicrobial potential of guava (Psidium guajava) leaf extracts against two gram-negative bacteria (Escherichia coli and Salmonella enteritidis) and two gram-positive bacteria (Staphylococcus aureus and Bacillus cereus) which are some of foodborne and spoilage bacteria. The guava leaves were extracted in four different solvents of increasing polarities (hexane, methanol, ethanol, and water). The efficacy of these extracts was tested against those bacteria through a well-diffusion method employing 50??L leaf-extract solution per well. According to the findings of the antibacterial assay, the methanol and ethanol extracts of the guava leaves showed inhibitory activity against gram-positive bacteria, whereas the gram-negative bacteria were resistant to all the solvent extracts. The methanol extract had an antibacterial activity with mean zones of inhibition of 8.27 and 12.3?mm, and the ethanol extract had a mean zone of inhibition of 6.11 and 11.0?mm against B. cereus and S. aureus, respectively. On the basis of the present finding, guava leaf-extract might be a good candidate in the search for a natural antimicrobial agent. This study provides scientific understanding to further determine the antimicrobial values and investigate other pharmacological properties.

Biswas, Bipul; Rogers, Kimberly; McLaughlin, Fredrick; Yadav, Anand

2013-01-01

131

Antimicrobial Activities of Leaf Extracts of Guava (Psidium guajava L.) on Two Gram-Negative and Gram-Positive Bacteria.  

PubMed

Aim. To determine the antimicrobial potential of guava (Psidium guajava) leaf extracts against two gram-negative bacteria (Escherichia coli and Salmonella enteritidis) and two gram-positive bacteria (Staphylococcus aureus and Bacillus cereus) which are some of foodborne and spoilage bacteria. The guava leaves were extracted in four different solvents of increasing polarities (hexane, methanol, ethanol, and water). The efficacy of these extracts was tested against those bacteria through a well-diffusion method employing 50? ? L leaf-extract solution per well. According to the findings of the antibacterial assay, the methanol and ethanol extracts of the guava leaves showed inhibitory activity against gram-positive bacteria, whereas the gram-negative bacteria were resistant to all the solvent extracts. The methanol extract had an antibacterial activity with mean zones of inhibition of 8.27 and 12.3?mm, and the ethanol extract had a mean zone of inhibition of 6.11 and 11.0?mm against B. cereus and S. aureus, respectively. On the basis of the present finding, guava leaf-extract might be a good candidate in the search for a natural antimicrobial agent. This study provides scientific understanding to further determine the antimicrobial values and investigate other pharmacological properties. PMID:24223039

Biswas, Bipul; Rogers, Kimberly; McLaughlin, Fredrick; Daniels, Dwayne; Yadav, Anand

2013-01-01

132

Phylogenetic diversity of Gram-positive bacteria cultured from Antarctic deep-sea sponges  

Microsoft Academic Search

Gram-positive bacteria, specifically actinobacteria and members of the order Bacillales, are well-known producers of important\\u000a secondary metabolites. Little is known about the diversity of Gram-positive bacteria associated with Antarctic deep-sea sponges.\\u000a In this study, cultivation-based approaches were applied to investigate the Gram-positive bacteria associated with the Antarctic\\u000a sponges Rossella nuda, Rossella racovitzae (Porifera: Hexactinellida), and Myxilla mollis, Homaxinella balfourensis, Radiella

Yanjuan Xin; Manmadhan Kanagasabhapathy; Dorte Janussen; Song Xue; Wei Zhang

133

Mechanisms of action of newer antibiotics for Gram-positive pathogens.  

PubMed

Certain Gram-positive bacteria, including meticillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and quinolone-resistant Streptococcus pneumoniae have achieved the status of "superbugs", in that there are few or no antibiotics available for therapy against these pathogens. Only a few classes of novel antibiotics have been introduced in the past 40 years, and all since 1999, including the streptogramin combination quinupristin/dalfopristin (Synercid), the oxazolidinone linezolid, and the lipopeptide daptomycin. This review discusses the mechanisms of antibiotic action against Gram-positive pathogens, and resistance counter-mechanisms developed by Gram-positive bacteria, with emphasis on the newer agents. PMID:15792738

Hancock, Robert Ew

2005-04-01

134

Rational Design of a Plasmid Origin That Replicates Efficiently in Both Gram-Positive and Gram-Negative Bacteria  

PubMed Central

Background Most plasmids replicate only within a particular genus or family. Methodology/Principal Findings Here we describe an engineered high copy number expression vector, pBAV1K-T5, that produces varying quantities of active reporter proteins in Escherichia coli, Acinetobacter baylyi ADP1, Agrobacterium tumefaciens, (all Gram-negative), Streptococcus pneumoniae, Leifsonia shinshuensis, Peanibacillus sp. S18-36 and Bacillus subtilis (Gram-positive). Conclusions/Significance Our results demonstrate the efficiency of pBAV1K-T5 replication in different bacterial species, thereby facilitating the study of proteins that don't fold well in E. coli and pathogens not amenable to existing genetic tools.

Bryksin, Anton V.; Matsumura, Ichiro

2010-01-01

135

Role of Encapsulation in the Pathogenesis of Anaerobic Gram-Positive Cocci.  

National Technical Information Service (NTIS)

The pathogenicity of 22 anaerobic and facultative Gram-positive cocci (AFGPC) was investigated by inoculating then into mice and determining their ability to cause subcutaneous abscesses. Only 11 heavily encapsulated isolates (> 50% of the cells were enca...

I. Brook R. I. Walker

1985-01-01

136

A Comparative Genome Analysis Identifies Distinct Sorting Pathways in Gram-Positive Bacteria  

Microsoft Academic Search

Surface proteins in gram-positive bacteria are frequently required for virulence, and many are attached to the cell wall by sortase enzymes. Bacteria frequently encode more than one sortase enzyme and an even larger number of potential sortase substrates that possess an LPXTG-type cell wall sorting signal. In order to elucidate the sorting pathways present in gram-positive bacteria, we performed a

David Comfort; Robert T. Clubb

2004-01-01

137

Structural Domain Based Multiple Instance Learning for Predicting Gram-Positive Bacterial Protein Subcellular Localization  

Microsoft Academic Search

Until recently, far few researches have been reported on Gram-positive protein subcelluar location prediction. Novel computational method is highly needed to help biologist design experiment. In this paper, we are motivated to propose a novel machine learning model for predicting Gram-positive protein subcelluar localization, as an alternative to the existing models Gpos-PLoc when the required GO annotation information is unavailable.

Suyu Mei; Wang Fei

2009-01-01

138

Protection from Lethal Gram-positive Infection by Macrophage Scavenger Receptor-dependent Phagocytosis  

Microsoft Academic Search

Infections with gram-positive bacteria are a major cause of morbidity and mortality in humans. Opsonin-dependent phagocytosis plays a major role in protection against and recovery from gram-positive infections. Inborn and acquired defects in opsonin generation and\\/or recognition by phagocytes are associated with an increased susceptibility to bacterial infections. In contrast, the physiological significance of opsonin-independent phagocytosis is unknown. Type I

Christian A. Thomas; Yongmei Li; Tatsuhiko Kodama; Hiroshi Suzuki; Samuel C. Silverstein; Joseph El Khoury

139

Antibiotic-Resistant Gram-Positive Cocci: Implications for Surgical Practice  

Microsoft Academic Search

. Gram-positive infections are causing more serious infections than ever before in surgical patients, who are increasingly\\u000a aged, ill, and debilitated. Invasive procedures disrupt natural barriers to bacterial invasion, and indwelling catheters may\\u000a act as conduits for infection. The use of broad-spectrum antibiotics selects for the emergence of resistant pathogens. Potential\\u000a sites of nosocomial gram-positive infections include the urinary tract,

Philip S. Barie

1998-01-01

140

Regulation of innate and adaptive immune responses by Gram-positive and Gram-negative bacteria  

Microsoft Academic Search

Bacteria are classified as Gram-positive or Gram-negative, depending on their cell wall structure. The role of the bacterial cell wall in immune regulation is the focus of the current work. Most Gram-positive bacteria stimulate monocytes to produce large amounts of IL-12. IL-12 induces production of IFN-? in T cells and NK cells, which, in turn, activates the bactericidal capacity of

Anna Martner

141

Transport Capabilities of Eleven Gram-positive Bacteria: Comparative Genomic Analyses  

PubMed Central

The genomes of eleven Gram-positive bacteria that are important for human health and the food industry, nine low G+C lactic acid bacteria and two high G+C Gram-positive organisms, were analyzed for their complement of genes encoding transport proteins. Thirteen to eighteen percent of their genes encode transport proteins, larger percentages than observed for most other bacteria. All of these bacteria possess channel proteins, some of which probably function to relieve osmotic stress. Amino acid uptake systems predominate over sugar and peptide cation symporters, and of the sugar uptake porters, those specific for oligosaccharides and glycosides often outnumber those for free sugars. About 10% of the total transport proteins are constituents of putative multidrug efflux pumps with Major Facilitator Superfamily (MFS)-type pumps (55%) being more prevalent than ATP-binding cassette (ABC)-type pumps (33%), which, however, usually greatly outnumber all other types. An exception to this generalization is Streptococcus thermophilus with 54% of its drug efflux pumps belonging to the ABC superfamily and 23% belonging each to the Multidrug/Oligosaccharide/Polysaccharide (MOP) superfamily and the MFS. These bacteria also display peptide efflux pumps that may function in intercellular signalling, and macromolecular efflux pumps, many of predictable specificities. Most of the bacteria analyzed have no pmf-coupled or transmembrane flow electron carriers. The one exception is Brevibacterium linens, which in addition to these carriers, also has transporters of several families not represented in the other ten bacteria examined. Comparisons with the genomes of organisms from other bacterial kingdoms revealed that lactic acid bacteria possess distinctive proportions of recognized transporter types (e.g., more porters specific for glycosides than reducing sugars). Some homologues of transporters identified had previously been identified only in Gram-negative bacteria or in eukaryotes. Our studies reveal unique characteristics of the lactic acid bacteria such as the universal presence of genes encoding mechanosensitive channels, competence systems and large numbers of sugar transporters of the phosphotransferase system. The analyses lead to important physiological predictions regarding the preferred signalling and metabolic activities of these industrially important bacteria.

Lorca, Graciela L.; Barabote, Ravi D.; Zlotopolski, Vladimir; Tran, Can; Winnen, Brit; Hvorup, Rikki N.; Stonestrom, Aaron J.; Nguyen, Elizabeth; Huang, Li-Wen; Kim, David S.; Saier, Milton H.

2007-01-01

142

Low-Molecular-Weight Protein Tyrosine Phosphatases of Bacillus subtilis  

Microsoft Academic Search

In gram-negative organisms, enzymes belonging to the low-molecular-weight protein tyrosine phosphatase (LMPTP) family are involved in the regulation of important physiological functions, including stress resistance and synthesis of the polysaccharide capsule. LMPTPs have been identified also in gram-positive bacteria, but their functions in these organisms are presently unknown. We cloned two putative LMPTPs from Bacillus subtilis, YfkJ and YwlE, which

Lucia Musumeci; Cristina Bongiorni; Lutz Tautz; Robert A. Edwards; Andrei Osterman; Marta Perego; Tomas Mustelin; Nunzio Bottini

2005-01-01

143

Development of a flow chart for identification of gram-positive anaerobic cocci in the clinical laboratory.  

PubMed

Gram-positive anaerobic cocci (GPAC) are a heterogeneous group of organisms that are isolated from clinical specimens more often than any group of anaerobic bacteria except Bacteroides species, yet many strains are still difficult or impossible to identify in the diagnostic laboratory. In this study, a total of 124 strains, including 13 reference strains of GPAC species and 111 isolates that had been recovered from clinical specimens previously and identified by 16S rRNA gene sequencing, were subjected to biochemical characterization. Based on the results, a short biochemical scheme that involves the minimum essential biochemical tests for accurate identification of clinically important GPAC was developed. PMID:17135439

Song, Yuli; Liu, Chengxu; Finegold, Sydney M

2007-02-01

144

Quinupristin-dalfopristin resistance among gram-positive bacteria in Taiwan.  

PubMed

To understand quinupristin-dalfopristin resistance among clinical isolates of gram-positive bacteria in Taiwan, where this agent is not yet available for clinical use, we evaluated 1,287 nonduplicate isolates recovered from January 1996 to December 1999 for in vitro susceptibility to quinupristin-dalfopristin and other newer antimicrobial agents. All methicillin-susceptible Staphylococcus aureus (MSSA) isolates were susceptible to quinupristin-dalfopristin. High rates of nonsusceptibility to quinupristin-dalfopristin (MICs, >/=2 microg/ml) were demonstrated for the following organisms: methicillin-resistant S. aureus (MRSA) (31%), coagulase-negative staphylococci (CoNS) (16%), Streptococcus pneumoniae (8%), viridans group streptococci (51%), vancomycin-susceptible enterococci (85%), vancomycin-resistant Enterococcus faecalis (100%), vancomycin-resistant Enterococcus faecium (66%), Leuconostoc spp. (100%), Lactobacillus spp. (50%), and Pediococcus spp. (87%). All isolates of MSSA, MRSA, S. pneumoniae, and viridans group streptococci were susceptible to vancomycin and teicoplanin. The rates of nonsusceptibility to vancomycin and teicoplanin were 5 and 7%, respectively, for CoNS, ranging from 12 and 18% for S. simulans to 0 and 0% for S. cohnii and S. auricularis. Moxifloxacin and trovafloxacin had good activities against these isolates except for ciprofloxacin-resistant vancomycin-resistant enterococci and methicillin-resistant staphylococci. In Taiwan, virginiamycin has been used in animal husbandry for more than 20 years, which may contribute to the high rates of quinupristin-dalfopristin resistance. PMID:11083643

Luh, K T; Hsueh, P R; Teng, L J; Pan, H J; Chen, Y C; Lu, J J; Wu, J J; Ho, S W

2000-12-01

145

Recovery of vancomycin-resistant gram-positive cocci from children.  

PubMed

A cross-sectional survey of vancomycin-resistant gram-positive cocci (VRGPC) in the feces of children was initiated after several bacteremic infections with these organisms occurred at our hospital. A selective medium consisting of colistin-nalidixic acid agar, 5% sheep blood, vancomycin (5 mg/liter), and amphotericin B (8 mg/liter) was developed to isolate VRGPC. A single stool specimen submitted to the clinical microbiology laboratory from each of 48 patients was inoculated onto the medium. Plates were incubated at 35 degrees C with 5% carbon dioxide and examined at 24, 48, and 72 h. Susceptibilities were determined by broth microdilution. A total of 14 isolates from 11 of 48 (22%) children were recovered. The density of growth ranged from a single colony to 2+. The VRGPC were identified as Leuconostoc lactis (n = 2), Lactobacillus confusus (n = 4), Enterococcus species (n = 5), and Lactococcus lactis (n = 3). One strain of Lactobacillus confusus was recovered from both the stool and the blood of one of these patients. The MICs of vancomycin were 4 micrograms/ml for one of the isolates, 8 micrograms/ml for four of the isolates, and more than 16 micrograms/ml for the remaining eight isolates. All isolates were susceptible to both penicillin and ampicillin. Only 1 of the 11 children had received prior treatment with vancomycin. We conclude that low concentrations of VRGPC may be common in the gastrointestinal tracts of children. PMID:2108993

Green, M; Wadowsky, R M; Barbadora, K

1990-03-01

146

Gram Positive Bacterial Superantigen Outside-In Signaling Causes Toxic Shock Syndrome  

PubMed Central

Staphylococcus aureus and Streptococcus pyogenes (group A streptococci) are gram-positive pathogens capable of producing a variety of bacterial exotoxins known as superantigens. Superantigens interact with antigen-presenting cells (APCs) and T cells to induce T cell proliferation and massive cytokine production, which leads to fever, rash, capillary leak, and subsequent hypotension, the major symptoms of toxic shock syndrome. Both S. aureus and group A streptococci colonize mucosal surfaces, including the anterior nares and vagina for S. aureus, and the oropharynx and less commonly the vagina for group A streptococci. However, due to their abilities to secrete a variety of virulence factors, the organisms can also cause illnesses from the mucosa. This review provides an updated discussion of the biochemical and structural features of one group of secreted virulence factors, the staphylococcal and group A streptococcal superantigens, and their abilities to cause toxic shock syndrome from a mucosal surface. The main focus of this review, however, is the abilities of superantigens to induce cytokines and chemokines from epithelial cells, which has been linked to a dodecapeptide region that is relatively conserved among all superantigens and is distinct from the binding sites required for interactions with APCs and T cells. This phenomenon, termed outside-in signaling, acts to recruit adaptive immune cells to the submucosa, where the superantigens can then interact with those cells to initiate the final cytokine cascades that lead to toxic shock syndrome.

Brosnahan, Amanda J.; Schlievert, Patrick M.

2011-01-01

147

Evaluation of the Verigene Gram-positive blood culture nucleic acid test for rapid detection of bacteria and resistance determinants.  

PubMed

Rapid identification of pathogens from blood cultures can decrease lengths of stay and improve patient outcomes. We evaluated the accuracy of the Verigene Gram-positive blood culture (BC-GP) nucleic acid test for investigational use only (Nanosphere, Inc., Northbrook, IL) for the identification of Gram-positive bacteria from blood cultures. The detection of resistance genes (mecA in Staphylococcus aureus and Staphylococcus epidermidis and vanA or vanB in Enterococcus faecium and Enterococcus faecalis) by the BC-GP assay also was assessed. A total of 186 positive blood cultures (in BacT/Alert FA bottles) with Gram-positive cocci observed with Gram staining were analyzed using the BC-GP assay. The BC-GP results were compared with the identification and susceptibility profiles obtained with routine methods in the clinical laboratory. Discordant results were arbitrated with additional biochemical, cefoxitin disk, and repeat BC-GP testing. The initial BC-GP organism identification was concordant with routine method results for 94.6% of the blood cultures. Only 40% of the Streptococcus pneumoniae identifications were correct. The detection of the mecA gene for 69 blood cultures with only S. aureus or S. epidermidis was concordant with susceptibility testing results. For 3 of 6 cultures with multiple Staphylococcus spp., mecA detection was reported but was correlated with oxacillin resistance in a species other than S. aureus or S. epidermidis. The detection of vanA agreed with susceptibility testing results for 45 of 46 cultures with E. faecalis or E. faecium. Comparison of the mean times to results for each organism group showed that BC-GP results were available 31 to 42 h earlier than phenotypic identifications and 41 to 50 h earlier than susceptibility results. PMID:23596240

Wojewoda, Christina M; Sercia, Linda; Navas, Maria; Tuohy, Marion; Wilson, Deborah; Hall, Geraldine S; Procop, Gary W; Richter, Sandra S

2013-07-01

148

Clinical update on linezolid in the treatment of Gram-positive bacterial infections  

PubMed Central

Gram-positive pathogens are a significant cause of morbidity and mortality in both community and health care settings. Glycopeptides have traditionally been the antibiotics of choice for multiresistant Gram-positive pathogens but there are problems with their use, including the emergence of glycopeptide-resistant strains, tissue penetration, and achieving and monitoring adequate serum levels. Newer antibiotics such as linezolid, a synthetic oxazolidinone, are available for the treatment of resistant Gram-positive bacteria. Linezolid is active against a wide range of Gram-positive bacteria and has been generally available for the treatment of Gram-positive infections since 2000. There are potential problems with linezolid use, including its bacteriostatic action and the relatively high incidence of reported adverse effects, particularly with long-term use. Long-term use may also be complicated by the development of resistance. However, linezolid has been shown to be clinically useful in the treatment of several serious infections where traditionally bacteriocidal agents have been required and many of its adverse effects are reversible on cessation. It has also been shown to be a cost-effective treatment option in several studies, with its high oral bioavailability allowing an early change from intravenous to oral formulations with consequent earlier patient discharge and lower inpatient costs.

Ager, Sally; Gould, Kate

2012-01-01

149

Vancomycin-resistant gram-positive bacteria isolated from human sources.  

PubMed Central

Recent reports of infections with vancomycin-resistant gram-positive bacteria prompted us to study vancomycin-resistant isolates from human sources to characterize the types of bacteria displaying this phenotype. Thirty-six vancomycin-resistant gram-positive isolates, 14 from clinical specimens and 22 from stool samples, were identified. These isolates were tentatively identified as Lactobacillus spp. (25 strains), Leuconostoc spp. (6 strains), and Pediococcus spp. (3 strains) on the basis of morphology and physiological tests. Two isolates of indeterminate morphology could not be unambiguously assigned to a genus. Four isolates of vancomycin-resistant lactobacilli from normally sterile body sites were considered to be clinically significant. Vancomycin-resistant gram-positive bacteria may represent an emerging class of nosocomial pathogens. Better methods for distinguishing the various genera in the clinical microbiology laboratory are needed.

Ruoff, K L; Kuritzkes, D R; Wolfson, J S; Ferraro, M J

1988-01-01

150

Interaction of Cationic Peptides with Lipoteichoic Acid and Gram-Positive Bacteria  

PubMed Central

Compounds with antiendotoxin properties have been extensively studied for their potential as therapeutic agents for sepsis attributable to gram-negative bacteria. However, with the increasing incidence of gram-positive sepsis, there is interest in identifying compounds with a broad spectrum of action against both gram-positive and gram-negative bacteria. A series of synthetic ?-helical cationic peptides related to bee melittin and silk moth cecropin have previously been shown to bind lipopolysaccharide (LPS) with high affinity, inhibit LPS-induced tumor necrosis factor alpha (TNF-?) production in vitro and in vivo, and kill gram-negative bacteria. In this study, we analyzed whether these peptides were active against gram-positive bacteria; whether they could bind to lipoteichoic acid (LTA), the major proinflammatory structure on gram-positive bacteria; and whether they could block the ability of LTA to promote the release of cytokines by the RAW 264.7 murine macrophage cell line. We found that the cationic peptides demonstrated moderate growth-inhibitory activity toward gram-positive bacteria. In addition, the peptides bound LTA with high affinity. This correlated with the ability of the peptides to block LTA-induced production of TNF and interleukin-6 by RAW 264.7 cells but did not correlate with their ability to kill the bacteria. The peptides also effectively inhibited LTA-induced TNF production in a whole human blood assay. The peptides were also able to partly block the ability of heat-killed Staphylococcus aureus, as well as soluble products of live S. aureus, to stimulate cytokine production by macrophages. Our results indicate that these cationic peptides may be useful to prevent sepsis and inflammation caused by both gram-negative and gram-positive bacteria.

Scott, Monisha G.; Gold, Michael R.; Hancock, Robert E. W.

1999-01-01

151

Complete genome sequence of the industrial bacterium Bacillus licheniformis and comparisons with closely related Bacillus species  

Microsoft Academic Search

Background  \\u000a Bacillus licheniformis is a Gram-positive, spore-forming soil bacterium that is used in the biotechnology industry to manufacture enzymes, antibiotics,\\u000a biochemicals and consumer products. This species is closely related to the well studied model organism Bacillus subtilis, and produces an assortment of extracellular enzymes that may contribute to nutrient cycling in nature.\\u000a \\u000a \\u000a \\u000a \\u000a Results  We determined the complete nucleotide sequence of the

Michael W Rey; Preethi Ramaiya; Beth A Nelson; Shari D Brody-Karpin; Elizabeth J Zaretsky; Maria Tang; Alfredo Lopez de Leon; Henry Xiang; Veronica Gusti; Ib Groth Clausen; Peter B Olsen; Michael D Rasmussen; Jens T Andersen; Per L Jørgensen; Thomas S Larsen; Alexei Sorokin; Alexander Bolotin; Alla Lapidus; Nathalie Galleron; S Dusko Ehrlich; Randy M Berka

2004-01-01

152

Paucisalibacillus globulus gen. nov., sp. nov., a Gram-positive bacterium isolated from potting soil.  

PubMed

A Gram-positive bacterium, designated B22(T), was isolated from potting soil produced in Portugal. This organism is a catalase-positive, oxidase-negative, motile, spore-forming, aerobic rod that grows optimally at 37 degrees C and pH 8.0-8.5. Optimal growth occurs in media containing 1 % (w/v) NaCl, although the organism can grow in 0-8 % NaCl. The cell wall peptidoglycan is of the A4alpha type with a cross-linkage containing d-Asp. The major respiratory quinone is menaquinone 7 and the major fatty acids are anteiso-15 : 0, anteiso-17 : 0 and iso-15 : 0. The DNA G+C content is 37.9 mol%. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain B22(T) formed a new branch within the family Bacillaceae. The novel isolate is phylogenetically closely related to members of genera of moderately halophilic bacilli and formed a coherent cluster with species of the genera Salinibacillus, Virgibacillus, Oceanobacillus and Lentibacillus, supported by bootstrap analysis at a confidence level of 71 %. Strain B22(T) exhibited 16S rRNA gene pairwise sequence similarity values of 94.7-94.3 % with members of the genus Salinibacillus, 95.1-92.8 % with members of the genus Virgibacillus, 94.7-93.2 % with members of the genus Oceanobacillus and 93.1-92.3 % with members of the genus Lentibacillus. On the basis of phylogenetic analysis and physiological and biochemical characteristics, it is proposed that strain B22(T) represents a novel species in a new genus, Paucisalibacillus globulus gen. nov., sp. nov. Strain B22(T) (=LMG 23148(T)=CIP 108857(T)) is the type strain of Paucisalibacillus globulus. PMID:16902018

Nunes, Inês; Tiago, Igor; Pires, Ana Luísa; da Costa, Milton S; Veríssimo, António

2006-08-01

153

Microarray-Based Detection of 90 Antibiotic Resistance Genes of Gram-Positive Bacteria  

Microsoft Academic Search

A disposable microarray was developed for detection of up to 90 antibiotic resistance genes in gram-positive bacteria by hybridization. Each antibiotic resistance gene is represented by two specific oligonucleotides chosen from consensus sequences of gene families, except for nine genes for which only one specific oligonucleotide could be developed. A total of 137 oligonucleotides (26 to 33 nucleotides in length

Vincent Perreten; Lorianne Vorlet-Fawer; Peter Slickers; Ralf Ehricht; Peter Kuhnert; Joachim Frey

2005-01-01

154

The Action of Lysozyme on Heat-killed Gram-positive Microorganisms  

Microsoft Academic Search

SUMMARY: The change in the Gram staining reaction which occurs when heat- killed Gram-positive Clostridium welchii and Staphylococcus albus are incubated with lysozyme is due to the removal of the ribonucleic acid component of the Gram complex, and is brought about by the hydrolysis of certain sugar linkages in polysaccharides located at the cell surface. Lysozyme, the enzyme in egg-white

M. Webb

1948-01-01

155

[Susceptibility of clinical strains of gram-positive bacteria to selected beta-lactam antibiotics].  

PubMed

The aim of the study was to determine the activity of four beta-lactam antibiotics against nosocomial strains of Gram-positive bacteria. Two antibiotics combined with beta-lactamase inhibitors: timentin (TIC/CLAV) and tazocin (PIP/TZB) and two carbapenems: imipenem and meropenem were applied. The clinical strains were isolated from patients hospitalized in surgical ward of the National Clinical Hospital No 1 in Warsaw. The strains were identified in the automatic ATB system using ID 32 STAPH, API STREP, API CORYNE and API 20 A strips. The susceptibility of isolates to antibacterial agents was determined in the automatic ATB system using ATB STAPH, ATB STREP and ATB ANA strips. The susceptibility of strains to timentin, tazocin, imipenem and meropenem was tested with disc diffusion method. 111 strains of Gram-positive bacteria were cultured. Staphylococci (49) and enterococci (44) dominated among isolated strains. 33 Staphylococcus spp. strains were identified as methicillin-resistant. The obtained results indicate a significant role of Gram-positive cocci (staphylococci and enterococci) in the aetiology of nosocomial infections. Antibiotics combined with beta-lactamase inhibitors and carbapenems demonstrate broad antibacterial spectrum against clinical strains of Gram-positive bacteria except E. faecium strains. PMID:9857616

Sawicka-Grzelak, A; Rokosz, A

1998-01-01

156

Gram-positive pathogenic bacteria induce a common early response in human monocytes  

Microsoft Academic Search

BACKGROUND: We infected freshly isolated human peripheral monocytes with live bacteria of three clinically important gram-positive bacterial species, Staphylococcus aureus, Streptococcus pneumoniae and Listeria monocytogenes and studied the ensuing early transcriptional response using expression microarrays. Thus the observed response was unbiased by signals originating from other helper and effector cells of the host and was not limited to induction by

Svetlin Tchatalbachev; Rohit Ghai; Hamid Hossain; Trinad Chakraborty

2010-01-01

157

Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria  

PubMed Central

Background The Gateway recombinatorial cloning system allows easy and rapid joining of DNA fragments. Here we report the construction and evaluation of three different Gram-positive vectors that can be used with the Multisite Gateway cloning system to rapidly produce new gene arrangements in plasmid constructs for use in a variety of Gram-positive bacteria. Results Comparison of patterns of reporter gene expression with conventionally constructed clones show that the presence of residual recombination (att) sites does not have an effect on patterns of gene expression, although overall levels of gene expression may vary. Rapid construction of these new vectors allowed vector/gene combinations to be optimized following evaluation of plasmid constructs in different bacterial cells and demonstrates the benefits of plasmid construction using Gateway cloning. Conclusion The residual att sites present after Gateway cloning did not affect patterns of promoter induction in Gram-positive bacteria and there was no evidence of differences in mRNA stability of transcripts. However overall levels of gene expression may be reduced, possibly due to some post-transcriptional event. The new vectors described here allow faster, more efficient cloning in range of Gram-positive bacteria.

Perehinec, Tania M; Qazi, Saara NA; Gaddipati, Sanyasi R; Salisbury, Vyvyan; Rees, Catherine ED; Hill, Philip J

2007-01-01

158

In vitro activity of daptomycin against Gram-positive cocci: the first multicentre study in Greece.  

PubMed

A total of 10420 Gram-positive cocci (including staphylococci, enterococci and various groups of streptococci) collected from clinically significant specimens in ten Greek hospitals during 2006--2007 were tested for their susceptibility to daptomycin. The minimum inhibitory concentration (MIC) was determined by the broth microdilution method. Daptomycin demonstrated very high activity against Enterococcus faecalis (MIC at which 50% of the isolates were inhibited (MIC50) = 1mg/L and MIC at which 90% of the isolates were inhibited (MIC90) = 1.36 mg/L), Enterococcus faecium (MIC50 = 1.36 mg/L and MIC90 = 1.90 mg/L), Streptococcus pyogenes (MIC50 = 0.12 mg/L and MIC90 = 0.50mg/L), Streptococcus agalactiae (MIC50 = 0.09 mg/L and MIC90 = 0.12 mg/L), Streptococcus pneumoniae (MIC50 = 0.24 mg/L and MIC90 = 0.5 mg/L) and viridans group streptococci (MIC50 = 0.50 mg/L and MIC90 = 0.89 mg/L). Resistance to linezolid and vancomycin for enterococci and to penicillin for streptococci appears to be independent of reduced susceptibility to daptomycin. On the other hand, daptomycin was also active against meticillin-resistant Staphylococcus aureus (MIC50 = 0.44 mg/L and MIC90 = 0.78 mg/L) and meticillin-resistant coagulase-negative staphylococci (MIC50 = 0.24 mg/L and MIC90 = 0.44 mg/L); however, 0.9% of the staphylococci tested had an MIC > 1mg/L, which is the Clinical and Laboratory Standards Institute breakpoint proposed for susceptibility. For all tested organism groups, resistance to daptomycin was not associated with glycopeptide resistance. PMID:18774268

Malli, E; Spiliopoulou, I; Kolonitsiou, F; Klapsa, D; Giannitsioti, E; Pantelidi, K; Pratti, A; Panopoulou, M; Grapsa, S; Alepopoulou, E; Neonakis, I; Frantzidou, F; Alexiou-Daniel, S; Bakola, D; Koutsia-Carouzou, C; Malamou-Lada, H; Zerva, L; Vlahaki, E; Kartali-Ktenidou, S; Anastassiou, E D; Petinaki, E

2008-12-01

159

Isolation and Characterization of Four Gram-Positive Nickel-Tolerant Microorganisms from Contaminated Sediments  

SciTech Connect

Microbial communities from riparian sediments contaminated with high levels of Ni and U were examined for metal-tolerant microorganisms. Isolation of four aerobic Ni-tolerant, Gram-positive heterotrophic bacteria indicated selection pressure from Ni. These isolates were identified as Arthrobacter oxydans NR-1, Streptomyces galbus NR-2, Streptomyces aureofaciens NR-3, and Kitasatospora cystarginea NR-4 based on partial 16S rDNA sequences. A functional gene microarray containing gene probes for functions associated with biogeochemical cycling, metal homeostasis, and organic contaminant degradation showed little overlap among the four isolates. Fifteen of the genes were detected in all four isolates with only two of these related to metal resistance, specifically to tellurium. Each of the four isolates also displayed resistance to at least one of six antibiotics tested, with resistance to kanamycin, gentamycin, and ciprofloxacin observed in at least two of the isolates. Further characterization of S. aureofaciens NR-3 and K. cystarginea NR-4 demonstrated that both isolates expressed Ni tolerance constitutively. In addition, both were able to grow in higher concentrations of Ni at pH 6 as compared with pH 7 (42.6 and 8.5 mM Ni at pH 6 and 7, respectively). Tolerance to Cd, Co, and Zn was also examined in these two isolates; a similar pH-dependent metal tolerance was observed when grown with Co and Zn. Neither isolate was tolerant to Cd. These findings suggest that Ni is exerting a selection pressure at this site for metal-resistant actinomycetes.

Van Nostrand, J. D.; Khijniak, T. V.; Gentry, T. J.; Novak, M. T.; Sowder, A. G.; Zhou, J. Z.; Bertsch, P. M.; Morris, P. J.

2007-01-01

160

Pulmonary innate immune proteins and receptors that interact with gram-positive bacterial ligands.  

PubMed

The two major gram-positive bacterial (GPB) ligands are peptidoglycan (PGN) and lipoteichoic acid (LTA). These polymeric LTA and highly organized PGN contain repeating carbohydrate moieties, which are potential targets for pattern recognition molecules. The major pattern recognition proteins and receptors, which bind GPB, either have a lectin, PGN recognition, collagen or leucine-rich repeat (LRR) domain. The soluble innate immune proteins (IIPs) that bind to PGN and LTA include pulmonary collectins surfactant-associated proteins (SP-) A and D, lectin-like pentraxins C-reactive protein (CRP) and serum amyloid P component (SAP), and sCD14. Membrane-anchored lectin or lectin-like group members include macrophage mannose receptor (MR), complement receptor 3 (CR3, or Mac-1, or integrin CD11b/CD18), scavenger receptor A (SRCL-1), lectin-like oxidized LDL receptor 1 (LOX-1), and GPI-anchored CD14. Although Toll-like receptor (TLR) 2 and 4, and CD14 contain extracellular LRR domains, only TLRs have a cytoplasmic domain for signal transduction. Three of the four recently discovered human PGN recognition proteins (PGRP) have a transmembrane domain, and hence, considered as true receptors for GPB. Since lysozyme is the only known pulmonary enzyme that can lyse bacterial cell wall PGN, other innate immune molecules appear to be responsible for signalling and enhancing the clearance of GPB infection from the lung. Interestingly, pulmonary collectins bind not only to GPB ligands but also to the receptors, CD14 and TLR, and antigen processing cells such as dentritic cells. These complex interactions appear to play major roles in linking innate and adaptive immunity, and maintaining a pathogen-free lung with minimal, or no inflammation. PMID:12396017

Palaniyar, Nades; Nadesalingam, Jeya; Reid, Kenneth B M

2002-09-01

161

Biosynthesis of Auxin by the Gram-Positive Phytopathogen Rhodococcus fascians Is Controlled by Compounds Specific to Infected Plant Tissues  

PubMed Central

The role and metabolism of indole-3-acetic acid in gram-negative bacteria is well documented, but little is known about indole-3-acetic acid biosynthesis and regulation in gram-positive bacteria. The phytopathogen Rhodococcus fascians, a gram-positive organism, incites diverse developmental alterations, such as leafy galls, on a wide range of plants. Phenotypic analysis of a leafy gall suggests that auxin may play an important role in the development of the symptoms. We show here for the first time that R. fascians produces and secretes the auxin indole-3-acetic acid. Interestingly, whereas noninfected-tobacco extracts have no effect, indole-3-acetic acid synthesis is highly induced in the presence of infected-tobacco extracts when tryptophan is not limiting. Indole-3-acetic acid production by a plasmid-free strain shows that the biosynthetic genes are located on the bacterial chromosome, although plasmid-encoded genes contribute to the kinetics and regulation of indole-3-acetic acid biosynthesis. The indole-3-acetic acid intermediates present in bacterial cells and secreted into the growth media show that the main biosynthetic route used by R. fascians is the indole-3-pyruvic acid pathway with a possible rate-limiting role for indole-3-ethanol. The relationship between indole-3-acetic acid production and the symptoms induced by R. fascians is discussed.

Vandeputte, Olivier; Oden, Sevgi; Mol, Adeline; Vereecke, Danny; Goethals, Koen; El Jaziri, Mondher; Prinsen, Els

2005-01-01

162

Biosynthesis of auxin by the gram-positive phytopathogen Rhodococcus fascians is controlled by compounds specific to infected plant tissues.  

PubMed

The role and metabolism of indole-3-acetic acid in gram-negative bacteria is well documented, but little is known about indole-3-acetic acid biosynthesis and regulation in gram-positive bacteria. The phytopathogen Rhodococcus fascians, a gram-positive organism, incites diverse developmental alterations, such as leafy galls, on a wide range of plants. Phenotypic analysis of a leafy gall suggests that auxin may play an important role in the development of the symptoms. We show here for the first time that R. fascians produces and secretes the auxin indole-3-acetic acid. Interestingly, whereas noninfected-tobacco extracts have no effect, indole-3-acetic acid synthesis is highly induced in the presence of infected-tobacco extracts when tryptophan is not limiting. Indole-3-acetic acid production by a plasmid-free strain shows that the biosynthetic genes are located on the bacterial chromosome, although plasmid-encoded genes contribute to the kinetics and regulation of indole-3-acetic acid biosynthesis. The indole-3-acetic acid intermediates present in bacterial cells and secreted into the growth media show that the main biosynthetic route used by R. fascians is the indole-3-pyruvic acid pathway with a possible rate-limiting role for indole-3-ethanol. The relationship between indole-3-acetic acid production and the symptoms induced by R. fascians is discussed. PMID:15746315

Vandeputte, Olivier; Oden, Sevgi; Mol, Adeline; Vereecke, Danny; Goethals, Koen; El Jaziri, Mondher; Prinsen, Els

2005-03-01

163

Comparative surveillance study of telavancin activity against recently collected gram-positive clinical isolates from across the United States.  

PubMed

Telavancin is an investigational, rapidly bactericidal lipoglycopeptide antibiotic that is being developed to treat serious infections caused by gram-positive bacteria. A baseline prospective surveillance study was conducted to assess telavancin activity, in comparison with other agents, against contemporary clinical isolates collected from 2004 to 2005 from across the United States. Nearly 4,000 isolates were collected, including staphylococci, enterococci, and streptococci (pneumococci, beta-hemolytic, and viridans). Telavancin had potent activity against Staphylococcus aureus and coagulase-negative staphylococci (MIC range, 0.03 to 1.0 microg/ml), independent of resistance to methicillin or to multiple agents. Telavancin activity was particularly potent against all streptococcal groups (MIC(90)s, 0.03 to 0.12 microg/ml). Telavancin had excellent activity against vancomycin-susceptible enterococci (MIC(90), 1 microg/ml) and was active against VanB strains of vancomycin-resistant enterococci (MIC(90), 2 microg/ml) but less active against VanA strains (MIC(90), 8 to 16 microg/ml). Telavancin also demonstrated activity against vancomycin-intermediate S. aureus and vancomycin-resistant S. aureus strains (MICs, 0.5 microg/ml to 1.0 microg/ml and 1.0 microg/ml to 4.0 microg/ml, respectively). These data may support the efficacy of telavancin for treatment of serious infections with a wide range of gram-positive organisms. PMID:18443115

Draghi, Deborah C; Benton, Bret M; Krause, Kevin M; Thornsberry, Clyde; Pillar, Chris; Sahm, Daniel F

2008-07-01

164

Comparative Surveillance Study of Telavancin Activity against Recently Collected Gram-Positive Clinical Isolates from across the United States ?  

PubMed Central

Telavancin is an investigational, rapidly bactericidal lipoglycopeptide antibiotic that is being developed to treat serious infections caused by gram-positive bacteria. A baseline prospective surveillance study was conducted to assess telavancin activity, in comparison with other agents, against contemporary clinical isolates collected from 2004 to 2005 from across the United States. Nearly 4,000 isolates were collected, including staphylococci, enterococci, and streptococci (pneumococci, beta-hemolytic, and viridans). Telavancin had potent activity against Staphylococcus aureus and coagulase-negative staphylococci (MIC range, 0.03 to 1.0 ?g/ml), independent of resistance to methicillin or to multiple agents. Telavancin activity was particularly potent against all streptococcal groups (MIC90s, 0.03 to 0.12 ?g/ml). Telavancin had excellent activity against vancomycin-susceptible enterococci (MIC90, 1 ?g/ml) and was active against VanB strains of vancomycin-resistant enterococci (MIC90, 2 ?g/ml) but less active against VanA strains (MIC90, 8 to 16 ?g/ml). Telavancin also demonstrated activity against vancomycin-intermediate S. aureus and vancomycin-resistant S. aureus strains (MICs, 0.5 ?g/ml to 1.0 ?g/ml and 1.0 ?g/ml to 4.0 ?g/ml, respectively). These data may support the efficacy of telavancin for treatment of serious infections with a wide range of gram-positive organisms.

Draghi, Deborah C.; Benton, Bret M.; Krause, Kevin M.; Thornsberry, Clyde; Pillar, Chris; Sahm, Daniel F.

2008-01-01

165

Atopococcus tabaci gen. nov., sp. nov., a novel Gram-positive, catalase-negative, coccus-shaped bacterium isolated from tobacco  

Microsoft Academic Search

A novel Gram-positive, aerobic, catalase-negative, coccus-shaped organism originating from tobacco was characterized using phenotypic and molecular taxonomic methods. The organism contained a cell wall murein based on L-lysine (variation A4a, type L-lysine-L-glutamic acid), synthesized long-chain cellular fatty acids of the straight-chain saturated and monounsaturated types (with C16 : 1v9, C16 : 0 and C18 : 1v9 predominating) and possessed a

Matthew D. Collins; Anna Wiernik; Enevold Falsen; Paul A. Lawson

2005-01-01

166

Reduction of Cr(VI) and bioaccumulation of chromium by gram positive and gram negative microorganisms not previously exposed to Cr-stress.  

PubMed

Resistance to Cr(VI) is usually associated with its cellular exclusion, precluding enrichment techniques for the isolation of organisms accumulating Cr(VI) via bioreduction to insoluble Cr(III). A technique was developed to screen for potential Cr(VI) reduction in approx. 2000 isolates from a coastal environment, based on the non-specific reduction of selenite and tellurite to Se0 and Te0, and reduction of tetrazolium blue to insoluble blue formazan. The most promising strains were further screened in liquid culture, giving three, which were identified by 16S rRNA sequence analysis as Bacillus pumilus, Exiguobacterium aurantiacum and Pseudomonas synxantha, all of which reduced 100 microM Cr(VI) anaerobically, without growth. The respective removal of Cr(VI) was 90% and 80% by B. pumilus and E. aurantiacum after 48 h and 80% and by P. synxantha after 192 h. With the gram positive strains Cr(VI) promoted loss of flagella and, in the case of B. pumilus, lysis of some cells, but Cr was deposited as an exocellular precipitate which was identified as containing Cr and P using energy dispersive X-ray microanalysis (EDAX). This prompted the testing of Citrobacter sp. N14 (subsequently re-assigned by 16S rRNA sequence analysis and biochemical studies as a strain of Serratia) which bioprecipitates metal cation phosphates via enzymatically-liberated phosphate. This strain reduced Cr(VI) at a rate comparable to that of P. synxantha but Cr(III) was not bioprecipitated where La(III) was removed as LaPO4, even though a similar amount of phosphate was produced in the presence of Cr(III). Since B. pumilus removed most of the Cr(VI), with the formation of cell-bound CrPO4 implicated, this suggests that this strain could have future bioprocess potential. PMID:12164635

Pattanapipitpaisal, P; Mabbett, A N; Finlay, J A; Beswick, A J; Paterson-Beedle, M; Essa, A; Wright, J; Tolley, M R; Badar, U; Ahmed, N; Hobman, J L; Brown, N L; Macaskie, L E

2002-07-01

167

Alternating electric fields combined with activated carbon for disinfection of Gram negative and Gram positive bacteria in fluidized bed electrode system.  

PubMed

Strong electric fields for disinfection of wastewaters have been employed already for several decades. An innovative approach combining low strength (7 V/cm) alternating electric fields with a granular activated carbon fluidized bed electrode (FBE) for disinfection was presented recently. For disinfection performance of FBE several pure microbial cultures were tested: Bacillus subtilis, Bacillus subtilis subsp. subtilis, Enterococcus faecalis as representatives from Gram positive bacteria and Erwinia carotovora, Pseudomonas luteola, Pseudomonas fluorescens and Escherichia coli YMc10 as representatives from Gram negative bacteria. The alternating electric field amplitude and shape were kept constant. Only the effect of alternating electric field frequency on disinfection performance was investigated. From the bacteria tested, the Gram negative strains were more susceptible and the Gram positive microorganisms were more resistant to FBE disinfection. The collected data indicate that the efficiency of disinfection is frequency and strain dependent. During 6 h of disinfection, the decrease above 2 Log units was achieved with P. luteola and E. coli at 10 kHz and at dual frequency shift keying (FSK) modulated signal with frequencies of 10 kHz and 140 kHz. FBE technology appears to offer a new way for selective bacterial disinfection, however further optimizations are needed on treatment duration, and energy input, to improve effectiveness. PMID:24012021

Racyte, Justina; Bernard, Séverine; Paulitsch-Fuchs, Astrid H; Yntema, Doekle R; Bruning, Harry; Rijnaarts, Huub H M

2013-10-15

168

The potent antimicrobial properties of cell penetrating peptide-conjugated silver nanoparticles with excellent selectivity for Gram-positive bacteria over erythrocytes  

NASA Astrophysics Data System (ADS)

Silver nanoparticles are of great interest for use as antimicrobial agents. Studies aimed at producing potent nano-silver biocides have focused on manipulation of particle size, shape, composition and surface charge. Here, we report the cell penetrating peptide catalyzed formation of antimicrobial silver nanoparticles in N,N-dimethylformamide. The novel nano-composite demonstrated a distinctly enhanced biocidal effect toward bacteria (Gram-positive Bacillus subtilis, Gram-negative Escherichia coli) and pathogenic yeast (Candida albicans), as compared to triangular and extremely small silver nanoparticles. In addition, a satisfactory biocompatibility was verified by a haemolysis test. Our results provide a paradigm in developing strategies that can maximize the silver nanoparticle application potentials while minimizing the toxic effects.Silver nanoparticles are of great interest for use as antimicrobial agents. Studies aimed at producing potent nano-silver biocides have focused on manipulation of particle size, shape, composition and surface charge. Here, we report the cell penetrating peptide catalyzed formation of antimicrobial silver nanoparticles in N,N-dimethylformamide. The novel nano-composite demonstrated a distinctly enhanced biocidal effect toward bacteria (Gram-positive Bacillus subtilis, Gram-negative Escherichia coli) and pathogenic yeast (Candida albicans), as compared to triangular and extremely small silver nanoparticles. In addition, a satisfactory biocompatibility was verified by a haemolysis test. Our results provide a paradigm in developing strategies that can maximize the silver nanoparticle application potentials while minimizing the toxic effects. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr34254a

Liu, Lihong; Yang, Jun; Xie, Jianping; Luo, Zhentao; Jiang, Jiang; Yang, Yi Yan; Liu, Shaomin

2013-04-01

169

Horizontal transfer of drug-resistant aminoacyl-transfer-RNA synthetases of anthrax and Gram-positive pathogens.  

PubMed

The screening of new antibiotics against several bacterial strains often reveals unexpected occurrences of natural drug resistance. Two examples of this involve specific inhibitors of Staphylococcus aureus isoleucyl-transfer-RNA synthetase 1 (IleRS1) and, more recently, Streptococcus pneumoniae methionyl-tRNA synthetase 1 (MetRS1). In both cases, resistance is due to the presence of a second gene that encodes another synthetase (IleRS2 or MetRS2). Here, we show that both S. pneumoniae MetRS2 and S. aureus IleRS2 have closely related homologues in the Gram-positive bacterium Bacillus anthracis, the causative agent of anthrax. Furthermore, similar to drug-resistant pathogens, strains of B. anthracis and its closest relative, B. cereus, also have wild-type ileS1 and metS1 genes. Clostridium perfringens, the causative agent of gangrene, also has two metS genes, whereas Oceanobacillus iheyensis isolated from deep-sea sediments has a single ileS2-type gene. This study shows the importance of understanding complex evolutionary networks of ancient horizontal gene transfer for the development of novel antibiotics. PMID:12792655

Brown, James R; Gentry, Daniel; Becker, Julie A; Ingraham, Karen; Holmes, David J; Stanhope, Michael J

2003-07-01

170

Sec-secretion and sortase-mediated anchoring of proteins in Gram-positive bacteria.  

PubMed

Signal peptide-driven secretion of precursor proteins directs polypeptides across the plasma membrane of bacteria. Two pathways, Sec- and SRP-dependent, converge at the SecYEG translocon to thread unfolded precursor proteins across the membrane, whereas folded preproteins are routed via the Tat secretion pathway. Gram-positive bacteria lack an outer membrane and are surrounded by a rigid layer of peptidoglycan. Interactions with their environment are mediated by proteins that are retained in the cell wall, often through covalent attachment to the peptidoglycan. In this review, we describe the mechanisms for both Sec-dependent secretion and sortase-dependent assembly of proteins in the envelope of Gram-positive bacteria. This article is part of a Special Issue entitled: Protein trafficking and secretion in bacteria. Guest Editors: Anastassios Economou and Ross Dalbey. PMID:24269844

Schneewind, Olaf; Missiakas, Dominique

2014-08-01

171

Novel Surface Display System for Proteins on Non-Genetically Modified Gram-Positive Bacteria  

PubMed Central

A novel display system is described that allows highly efficient immobilization of heterologous proteins on bacterial surfaces in applications for which the use of genetically modified bacteria is less desirable. This system is based on nonliving and non-genetically modified gram-positive bacterial cells, designated gram-positive enhancer matrix (GEM) particles, which are used as substrates to bind externally added heterologous proteins by means of a high-affinity binding domain. This binding domain, the protein anchor (PA), was derived from the Lactococcus lactis peptidoglycan hydrolase AcmA. GEM particles were typically prepared from the innocuous bacterium L. lactis, and various parameters for the optimal preparation of GEM particles and binding of PA fusion proteins were determined. The versatility and flexibility of the display and delivery technology were demonstrated by investigating enzyme immobilization and nasal vaccine applications.

Bosma, Tjibbe; Kanninga, Rolf; Neef, Jolanda; Audouy, Sandrine A. L.; van Roosmalen, Maarten L.; Steen, Anton; Buist, Girbe; Kok, Jan; Kuipers, Oscar P.; Robillard, George; Leenhouts, Kees

2006-01-01

172

Ubiquitous Detection of Gram-Positive Bacteria with Bioorthogonal Magnetofluorescent Nanoparticles  

PubMed Central

The ability to rapidly diagnose gram-positive pathogenic bacteria would have far reaching biomedical and technological applications. Here we describe the bioorthogonal modification of small molecule antibiotics (vancomycin and daptomycin), which bind to the cell wall of gram-positive bacteria. The bound antibiotics conjugates can be reacted orthogonally with tetrazine-modified nanoparticles, via an almost instantaneous cycloaddition, which subsequently renders the bacteria detectable by optical or magnetic sensing. We show that this approach is specific, selective, fast and biocompatible. Furthermore, it can be adapted to the detection of intracellular pathogens. Importantly, this strategy enables detection of entire classes of bacteria, a feat that is difficult to achieve using current antibody approaches. Compared to covalent nanoparticle conjugates, our bioorthogonal method demonstrated 1-2 orders of magnitude greater sensitivity. This bioorthogonal labeling method could ultimately be applied to a variety of other small molecules with specificity for infectious pathogens, enabling their detection and diagnosis.

Chung, Hyun Jung; Reiner, Thomas; Budin, Ghyslain; Min, Changwook; Liong, Monty; Issadore, David; Lee, Hakho; Weissleder, Ralph

2011-01-01

173

Lipoteichoic acids, phosphate-containing polymers in the envelope of gram-positive bacteria.  

PubMed

Lipoteichoic acids (LTA) are polymers of alternating units of a polyhydroxy alkane, including glycerol and ribitol, and phosphoric acid, joined to form phosphodiester units that are found in the envelope of Gram-positive bacteria. Here we review four different types of LTA that can be distinguished on the basis of their chemical structure and describe recent advances in the biosynthesis pathway for type I LTA, d-alanylated polyglycerol-phosphate linked to di-glucosyl-diacylglycerol. The physiological functions of type I LTA are discussed in the context of inhibitors that block their synthesis and of mutants with discrete synthesis defects. Research on LTA structure and function represents a large frontier that has been investigated in only few Gram-positive bacteria. PMID:24415723

Schneewind, Olaf; Missiakas, Dominique

2014-03-01

174

Isolation and Characterization of Four Gram-Positive Nickel-Tolerant Microorganisms from Contaminated Sediments  

Microsoft Academic Search

Microbial communities from riparian sediments contaminated with high levels of Ni and U were examined for metal-tolerant microorganisms.\\u000a Isolation of four aerobic Ni-tolerant, Gram-positive heterotrophic bacteria indicated selection pressure from Ni. These isolates\\u000a were identified as Arthrobacter oxydans NR-1, Streptomyces galbus NR-2, Streptomyces aureofaciens NR-3, and Kitasatospora cystarginea NR-4 based on partial 16S rDNA sequences. A functional gene microarray containing

Joy D. Van Nostrand; Tatiana V. Khijniak; Terry J. Gentry; Michelle T. Novak; Andrew G. Sowder; Jizhong Z. Zhou; Paul M. Bertsch; Pamela J. Morris

2007-01-01

175

Plasmid vectors for Gram-positive bacteria switching from high to low copy number  

Microsoft Academic Search

A set of vectors for Gram-positive bacteria was constructed with a new feature which enables the switching down of their copy number per cell. These vectors carry the replication region of pAM?1, containing a gene essential for replication, repE, and its regulator, copF The latter gene was inactivated by inserting a linker into its unique KpnI site. Since copF downregulates

Pierre Renault; Gerard Corthier; Nathalie Goupil; Christine Delorme; S. Dusko Ehrlich

1996-01-01

176

Teichoic acids and related cell-wall glycopolymers in Gram-positive physiology and host interactions  

Microsoft Academic Search

Abstract | Most Gram-positive bacteria incorporate membrane- or peptidoglycan-attached carbohydrate-based polymers into their cell envelopes. Such cell-wall glycopolymers (CWGs) often have highly variable structures and have crucial roles in protecting, connecting and controlling the major envelope constituents. Further important roles of CWGs in host-cell adhesion, inflammation and immune activation have also been described in recent years. Identifying and harnessing highly

Christopher Weidenmaier; Andreas Peschel

2008-01-01

177

Vancomycin-resistant Gram-positive cocci: risk factors for faecal carriage  

Microsoft Academic Search

This case-control study was undertaken to identify the risk factors for the gastrointestinal carriage of vancomycin-resistant, Gram-positive cocci (VRGPC) including vancomycin-resistant enterococci (VRE). Use of oral vancomycin (P = 0·003) or cephalosporins (P = 0·03) and prolonged duration of stay in the hospital (P = 0·02) were found to be the significant risk factors. Other previously suggested risk factors such

G. Gopal Rao; F. Ojo; D. Kolokithas

1997-01-01

178

Cell to cell communication by autoinducing peptides in gram-positive bacteria  

Microsoft Academic Search

While intercellular communication systems in Gram-negative bacteria are often based on homoserine lactones as signalling molecules,\\u000a it has been shown that autoinducing peptides are involved in intercellular communication in Gram-positive bacteria. Many of\\u000a these peptides are exported by dedicated systems, posttranslationally modified in various ways, and finally sensed by other\\u000a cells via membrane-located receptors that are part of two-component regulatory

Mark H. J. Sturme; Michiel Kleerebezem; Jiro Nakayama; Antoon D. L. Akkermans; Elaine E. Vaughan; Willem M. de Vos

2002-01-01

179

Genome Sequence of the Diazotrophic Gram-Positive Rhizobacterium Paenibacillus riograndensis SBR5T  

PubMed Central

Paenibacillus riograndensis SBR5T, a nitrogen-fixing Gram-positive rhizobacterium isolated from a wheat field in the south of Brazil, has a great potential for agricultural applications due to its plant growth promotion effects. Here we present the draft genome sequence of P. riograndensis SBR5T. Its 7.37-Mb genome encodes determinants of the diazotrophic lifestyle and plant growth promotion, such as nitrogen fixation, antibiotic resistance, nitrate utilization, and iron uptake.

Beneduzi, Anelise; Campos, Samanta; Ambrosini, Adriana; de Souza, Rocheli; Granada, Camille; Costa, Pedro; Arruda, Leticia; Moreira, Fernanda; Vargas, Luciano K.; Weiss, Vinicius; Tieppo, Eduardo; Faoro, Helisson; de Souza, Emanuel M.; Pedrosa, Fabio O.; Passaglia, Luciane M. P.

2011-01-01

180

Drosophila Toll is activated by Gram-positive bacteria through a circulating peptidoglycan recognition protein  

Microsoft Academic Search

Microbial infection activates two distinct intracellular signalling cascades in the immune-responsive fat body of Drosophila. Gram-positive bacteria and fungi predominantly induce the Toll signalling pathway, whereas Gram-negative bacteria activate the Imd pathway. Loss-of-function mutants in either pathway reduce the resistance to corresponding infections. Genetic screens have identified a range of genes involved in these intracellular signalling cascades, but how they

Tatiana Michel; Jean-Marc Reichhart; Jules A. Hoffmann; Julien Royet

2001-01-01

181

On-Probe Sample Pretreatment for Direct Analysis of Lipids in Gram-Positive Bacterial Cells by Matrix-Assisted Laser Desorption Ionization Mass Spectrometry  

Microsoft Academic Search

On-probe sample pretreatment using trifluoroacetic acid as an additional reagent enabled the direct detection of phospholipids in whole bacteria by means of matrix-assisted laser desorption ionization mass spectrometry for not only gram-negative organisms but also gram-positive ones with a thicker peptidoglycan layer. Matrix-assisted laser desorption ionization mass spectrome- try (MALDI-MS) has become a powerful method to charac- terize lipid components

Yasuyuki Ishida; Kuniyuki Kitagawa; Akihito Nakayama; Hajime Ohtani

2005-01-01

182

Cell lysis and DNA extraction of gram-positive and gram-negative bacteria from whole blood in a disposable microfluidic chip  

Microsoft Academic Search

Sepsis caused by gram positive and gram negative bacteria is the leading cause of death in noncoronary ICUs and the tenth leading cause of death in the United States. We have developed a microfluidic sample preparation platform for rapid on-chip detection of infectious organisms for point-of-care diagnostics. The microfluidic chips are made of a robust thermoplastic and can be easily

Madhumita Mahalanabis; Hussam Al-Muayad; M. Dominika Kulinski; Dave Altmanb; Catherine M. Klapperichac

2009-01-01

183

Isolation and Characterization of Microsphaera multipartita gen. nov., sp. nov., a Polysaccharide-Accumulating Gram-Positive Bacterium from Activated Sludge  

Microsoft Academic Search

A new gram-positive bacterium was isolated from activated sludge acclimated with sugar-containing syn- thetic wastewater. This organism, designated strain Y-104T (T = type strain), was a coccus-shaped, aerobic chemoorganotroph that had a strictly respiratory type of metabolism with oxygen as the terminal electron acceptor. This strain accumulates large amounts of polysaccharide in its cells. Strain Y-104T has the following chemotaxonomic

YUKIHIKO YOSHIM; AURA HIRAISHI; KAZUNORI NAKAMUFU

184

Platelet and Neutrophil Responses to Gram Positive Pathogens in Patients with Bacteremic Infection  

PubMed Central

Background Many Gram-positive pathogens aggregate and activate platelets in vitro and this has been proposed to contribute to virulence. Platelets can also form complexes with neutrophils but little is however known about platelet and platelet-neutrophil responses in bacterial infection. Methodology/Principal Findings We added isolates of Gram-positive bacteria from 38 patients with a bacteremic infection to blood drawn from the same patient. Aggregometry and flow cytometry were used to assess platelet aggregation and to quantify activation of platelets, neutrophils, and platelet-neutrophils complexes (PNCs) induced by the bacteria. Fifteen healthy persons served as controls. Most isolates of Staphylococcus aureus, beta hemolytic streptococci, and Enterococcus faecalis induced aggregation of platelets from their respective hosts, whereas pneumococci failed to do so. S. aureus isolates induced platelet aggregation more rapidly in patients than in controls, whereas platelet activation by S. aureus was lower in patients than in controls. PNCs were more abundant in baseline samples from patients than in healthy controls and most bacterial isolates induced additional PNC formation and neutrophil activation. Conclusion/Significance We have demonstrated for the first time that bacteria isolated from patients with Gram-positive bacteremia can induce platelet activation and aggregation, PNC formation, and neutrophil activation in the same infected host. This underlines the significance of these interactions during infection, which could be a target for future therapies in sepsis.

Johansson, Daniel; Shannon, Oonagh; Rasmussen, Magnus

2011-01-01

185

Multiple Responses of Gram-Positive and Gram-Negative Bacteria to Mixture of Hydrocarbons  

PubMed Central

Most of our knowledge about pollutants and the way they are biodegraded in the environment has previously been shaped by laboratory studies using hydrocarbon-degrading bacterial strains isolated from polluted sites. In present study Gram-positive (Mycobacterium sp. IBBPo1, Oerskovia sp. IBBPo2, Corynebacterium sp. IBBPo3) and Gram-negative (Chryseomonas sp. IBBPo7, Pseudomonas sp. IBBPo10, Burkholderia sp. IBBPo12) bacteria, isolated from oily sludge, were found to be able to tolerate pure and mixture of saturated hydrocarbons, as well as pure and mixture of monoaromatic and polyaromatic hydrocarbons. Isolated Gram-negative bacteria were more tolerant to mixture of saturated (n-hexane, n-hexadecane, cyclohexane), monoaromatic (benzene, toluene, ethylbenzene) and polyaromatic (naphthalene, 2-methylnaphthalene, fluorene) hydrocarbons than Gram-positive bacteria. There were observed cellular and molecular modifications induced by mixture of saturated, monoaromatic and polyaromatic hydrocarbons to Gram-positive and Gram-negative bacteria. These modifications differ from one strain to another and even for the same bacterial strain, according to the nature of hydrophobic substrate.

Marilena Lazaroaie, Mihaela

2010-01-01

186

A Comparative Genome Analysis Identifies Distinct Sorting Pathways in Gram-Positive Bacteria  

PubMed Central

Surface proteins in gram-positive bacteria are frequently required for virulence, and many are attached to the cell wall by sortase enzymes. Bacteria frequently encode more than one sortase enzyme and an even larger number of potential sortase substrates that possess an LPXTG-type cell wall sorting signal. In order to elucidate the sorting pathways present in gram-positive bacteria, we performed a comparative analysis of 72 sequenced microbial genomes. We show that sortase enzymes can be partitioned into five distinct subfamilies based upon their primary sequences and that most of their substrates can be predicted by making a few conservative assumptions. Most bacteria encode sortases from two or more subfamilies, which are predicted to function nonredundantly in sorting proteins to the cell surface. Only ?20% of sortase-related proteins are most closely related to the well-characterized Staphylococcus aureus SrtA protein, but nonetheless, these proteins are responsible for anchoring the majority of surface proteins in gram-positive bacteria. In contrast, most sortase-like proteins are predicted to play a more specialized role, with each anchoring far fewer proteins that contain unusual sequence motifs. The functional sortase-substrate linkage predictions are available online (http://www.doe-mbi.ucla.edu/Services/Sortase/) in a searchable database.

Comfort, David; Clubb, Robert T.

2004-01-01

187

Benfang Lei's research on heme acquisition in Gram-positive pathogens and bacterial pathogenesis  

PubMed Central

Benfang Lei’s laboratory conducts research on pathogenesis of human pathogen Group A Streptococcus (GAS) and horse pathogen Streptococcus equi (S. equi). His current research focuses on heme acquisition in Gram-positive pathogens and molecular mechanism of GAS and S. equi pathogenesis. Heme is an important source of essential iron for bacterial pathogens. Benfang Lei and colleagues identified the first cell surface heme-binding protein in Gram-positive pathogens and the heme acquisition system in GAS, demonstrated direct heme transfer from one protein to another, demonstrated an experimental pathway of heme acquisition by the Staphylococcus aureus Isd system, elucidated the activated heme transfer mechanism, and obtained evidence for a chemical mechanism of direct axial ligand displacement during the Shp-to-HtsA heme transfer reaction. These findings have considerably contributed to the progress that has been made over recent years in understanding the heme acquisition process in Gram-positive pathogens. Pathogenesis of GAS is mediated by an abundance of extracellular proteins, and pathogenic role and functional mechanism are not known for many of these virulence factors. Lei laboratory identified a secreted protein of GAS as a CovRS-regulated virulence factor that is a protective antigen and is critical for GAS spreading in the skin and systemic dissemination. These studies may lead to development of novel strategies to prevent and treat GAS infections.

Lei, Benfang

2010-01-01

188

Electrochemical classification of gram-negative and gram-positive bacteria.  

PubMed Central

Intestinal bacteria were classified as gram-positive or gram-negative by an electrode system with a basal plane pyrolytic graphite electrode and a porous nitrocellulose membrane filter to trap bacteria. When the potential of the graphite electrode was run in the range of 0 to 1.0 V versus the saturated calomel electrode (SCE), gram-positive bacteria gave peak currents at 0.65 to 0.69 V versus the SCE. The peak potentials of gram-negative bacteria were 0.70 to 0.74 V versus the SCE. Gram-negative bacteria and gram-positive bacteria were also classified based on the ratio of the second peak current to the first peak current when the potential cycle was repeated twice. The numbers of cells on the membrane filter were determined from the peak currents. It was found that the peak currents result from the electrochemical oxidation of coenzyme A in the cells of Escherichia coli and Lactobacillus acidophilus. Images

Matsunaga, T; Nakajima, T

1985-01-01

189

Surface Proteins of Gram-Positive Pathogens: Using Crystallography to Uncover Novel Features in Drug and Vaccine Candidates  

NASA Astrophysics Data System (ADS)

Proteins displayed on the cell surfaces of pathogenic organisms are the front-line troops of bacterial attack, playing critical roles in colonization, infection and virulence. Although such proteins can often be recognized from genome sequence data, through characteristic sequence motifs, their functions are often unknown. One such group of surface proteins is attached to the cell surface of Gram-positive pathogens through the action of sortase enzymes. Some of these proteins are now known to form pili: long filamentous structures that mediate attachment to human cells. Crystallographic analyses of these and other cell surface proteins have uncovered novel features in their structure, assembly and stability, including the presence of inter- and intramolecular isopeptide crosslinks. This improved understanding of structures on the bacterial cell surface offers opportunities for the development of some new drug targets and for novel approaches to vaccine design.

Baker, Edward N.; Proft, Thomas; Kang, Haejoo

190

[MRSA/MRSE-VISA/GISA/VRSA-PRP-VRE: current gram positive problem bacteria and mechanism of resistance, prevalence and clinical consequences].  

PubMed

The raising frequency of occurrence of multi-resistant Gram-positive pathogens has led to difficult to treat infections not only in hospitals but also in outpatient settings. Methicillin-resistant Staphylococcus aureus, the glycopeptide-susceptibility of which one cannot be sure of any more, vancomycin-resistant enterococci and penicillin-resistant pneumococci are on the top of the list of Gram-positive problem organisms nowadays. Clinical microbiology laboratories are faced with the challenge of accurately detecting emerging antibiotic resistance, which might be difficult and purpose of reporting data should be twofold: First to report adequately identified, relevant organisms as well as their susceptibility profiles to clinicians in context with adequate patient management and secondly to report continuously epidemiological data in form of statistics to the community in general and within a given setting as could be a specified hospital in particular. For reliable detection, laboratories may need to employ special screening- and susceptibility testing methods. Certain of these tests are highly specific, while others may require additional confirmatory testing for definite results. This article reports on current resistance mechanisms of gram-positive pathogens and subsequently resulting consequences for clinicians. Moreover, the frequency of occurrence in Austria in general as well as at the Vienna General Hospital in particular will be referred to. PMID:12764866

Apfalter, Petra

2003-01-01

191

Differential Expression of Two Paralogous Genes of Bacillus subtilis Encoding Single-Stranded DNA Binding Protein  

Microsoft Academic Search

The Bacillus subtilis genome comprises two paralogous single-stranded DNA binding protein (SSB) genes, ssb and ywpH, which show distinct expression patterns. The main ssb gene is strongly expressed during exponential growth and is coregulated with genes encoding the ribosomal proteins S6 and S18. The gene organization rpsF-ssb-rpsR as observed in B. subtilis is found in many gram-positive as well as

Cordula Lindner; Reindert Nijland; Mariska van Hartskamp; Sierd Bron; Leendert W. Hamoen; Oscar P. Kuipers

2004-01-01

192

Homologous Recombination in Low dC + dG Gram-Positive Bacteria  

Microsoft Academic Search

Homologous recombination is a process involved in the maintenance of chromosome integrity,\\u000a in shaping the evolution of pathogens, in the resistance to antibiotic treatment, and profoundly affecting\\u000a evolution. In low dC + dG Gram-positive bacteria genetic recombination of a non-replicative\\u000a \\u000a homologous DNA, which enters into the cell via transduction or conjugation, proceeds mainly by the\\u000a double-strand break repair machinery, and this process

Humberto Sanchez; Begoña Carrasco; Silvia Ayora; Juan C. Alonso

193

Structure-Activity Relationships of 3,3?-Phenylmethylene-bis-4-hydroxycoumarins: Selective and Potent Inhibitors of Gram-Positive Bacteria  

PubMed Central

Dicoumarols and coumarin derivatives have shown a variety of pharmaceutical activities and have been found to be potent inhibitor for the NAD(P)H-dependent flavoproteins. In this report, dicoumarol and its derivatives containing the substituted benzene ring at the methylenebis position were synthesized and evaluated for their antibacterial activity against gram-positive bacteria: Staphylococcus aureus and Bacillus subtilis, and gram-negative bacteria: Escherichia coli and Klebsiella sp. The results showed that the synthesized dicoumarols affect cell growth but are selective against gram-positive over gram-negative bacterial cells. However, for most derivatives, the substitution of steric bulky benzene group on the methylenebis position appears to decrease in the efficacy of antibacterial effect. This finding is roughly described by the predicted poorer docked structure of the derivatives to a homology model of S. aureus flavoprotein. 3D-QSAR study highlighted structural features around the substituted benzene ring of dicoumarols as the antibacterial activity. CoMFA and CoMSIA contour maps support the idea that steric repulsion at the para position could diminish the antibacterial activity. The results of this study provide a better understanding of the molecular basis for the antibacterial activity of dicoumarols.

Chavasiri, Warinthorn

2013-01-01

194

Agreement Between Deoxyribonucleic Acid Base Composition and Taxometric Classification of Gram-Positive Cocci.  

PubMed

Silvestri, L. G. (Università Statale, Milan, Italy), and L. R. Hill. Agreement between deoxyribonucleic acid base composition and taxometric classification of gram-positive cocci. J. Bacteriol. 90:136-140. 1965.-It had been previously proposed, from taxometric analyses, that gram-positive, catalase-positive cocci be divided into two subgroups. Thirteen strains, representative of both subgroups, were examined for deoxyribonucleic acid (DNA) base composition, determined from melting temperatures. Per cent GC (guanine + cytosine/total bases) values fell into two groups: 30.8 to 36.5% GC and 69 to 75% GC. Strains with low per cent GC values belonged to the Staphylococcus aureus-S. saprophyticus-S. lactis taxometric subgroups, and those with high per cent GC values belonged to the S. roseus-S. afermentans subgroup. The hypothetical nature of any classification is emphasized, and, in the present work, the hypothesis derived from taxometric analyses of division into two subgroups is confirmed by the study of DNA base ratios. The two subgroups correspond, respectively, to the genera Staphylococcus and Micrococcus. PMID:16562008

Silvestri, L G; Hill, L R

1965-07-01

195

Bioengineered nisin A derivatives with enhanced activity against both Gram positive and Gram negative pathogens.  

PubMed

Nisin is a bacteriocin widely utilized in more than 50 countries as a safe and natural antibacterial food preservative. It is the most extensively studied bacteriocin, having undergone decades of bioengineering with a view to improving function and physicochemical properties. The discovery of novel nisin variants with enhanced activity against clinical and foodborne pathogens has recently been described. We screened a randomized bank of nisin A producers and identified a variant with a serine to glycine change at position 29 (S29G), with enhanced efficacy against S. aureus SA113. Using a site-saturation mutagenesis approach we generated three more derivatives (S29A, S29D and S29E) with enhanced activity against a range of Gram positive drug resistant clinical, veterinary and food pathogens. In addition, a number of the nisin S29 derivatives displayed superior antimicrobial activity to nisin A when assessed against a range of Gram negative food-associated pathogens, including E. coli, Salmonella enterica serovar Typhimurium and Cronobacter sakazakii. This is the first report of derivatives of nisin, or indeed any lantibiotic, with enhanced antimicrobial activity against both Gram positive and Gram negative bacteria. PMID:23056510

Field, Des; Begley, Maire; O'Connor, Paula M; Daly, Karen M; Hugenholtz, Floor; Cotter, Paul D; Hill, Colin; Ross, R Paul

2012-01-01

196

Gene homogeneity for aminoglycoside-modifying enzymes in gram-positive cocci.  

PubMed Central

Aminoglycoside-resistant strains of Staphylococcus and Enterococcus, approximately 500 of each, were screened by dot blot hybridization for the presence of genes encoding aminoglycoside-modifying enzymes. The MICs of various aminoglycosides for the strains were determined, and the enzyme contents of the cells were inferred from the resistance phenotypes. The agreements (in percent) of the hybridization results with the deduced enzyme contents for Staphylococcus and Enterococcus species were, respectively, 80 and 87.6 for ANT(6) (aminoglycoside nucleotidyltransferase), 99.8 and 100 for both APH(3') (aminoglycoside phosphotransferase) and APH(2")-AAC(6') (aminoglycoside acetyltransferase), and 100 and 100 for ANT(4'). The weak correlation obtained with the probe for ANT(6) was due to the fact that gram-positive cocci can also be streptomycin resistant by synthesis of APH(3") or ANT(3")(9) and by ribosomal mutation. The remaining probes appeared to be specific: they hybridized with all the resistant clinical isolates but not with the susceptible strains. These results indicate that, except for streptomycin, nucleic acid hybridization is a valid approach for the detection and characterization of aminoglycoside resistance in gram-positive cocci.

Ounissi, H; Derlot, E; Carlier, C; Courvalin, P

1990-01-01

197

Overview of resistant gram-positive pathogens in the surgical patient.  

PubMed

Staphylococci and enterococci are the most common pathogens in surgical-site and bloodstream infections. The emergence of drug resistance among these gram-positive bacteria thus poses a substantial threat to patients with surgical infections. Resistance to methicillin/oxacillin is frequently observed in Staphylococcus aureus isolates and is often accompanied by multidrug resistance. Vancomycin is usually the treatment of choice for infections caused by methicillin-resistant S. aureus (MRSA), so the recent appearance of S. aureus isolated with intermediate sensitivity to vancomycin is cause for concern. Vancomycin resistance has already appeared in most species of enterococci. Infections caused by vancomycin-resistant enterococci (VRE) are associated with increased mortality compared to infections caused by vancomycin-sensitive isolates. Measures for preventing vancomycin resistance include reducing the use of vancomycin and other agents that appear to be associated with VRE, including third-generation cephalosporins and anti-anaerobic drugs. Third-generation cephalosporins have also been implicated in the increased prevalence of MRSA infections. Prudent use of existing antibiotics is an essential strategy for combating the rising tide of drug-resistant gram-positive pathogens. PMID:12594908

Rapp, R P

2000-01-01

198

Fluorescence studies of gram-positive and gram-negative bacteria  

NASA Astrophysics Data System (ADS)

Autofluorescence is a relatively unexplored technique for identification. It is nondestructive, noncontact, fast, and has the potential to be integrated in small handheld devices. On the other hand, the autofluorescent signal is sometimes very week, or it can be overwhelmed by the emission of a surrounding medium. We are exploring the possibility to develop an optical method for identification of the Gram-type of bacterial cultures based on the autofluorescence. We have enhanced the detectivity of a standard fluorimeter using combination of bandpass and long pass filters. In this particular study, we are investigating if the previously observed difference in the autofluorescent spectra of Gram-positive and Gram-negative bacteria is dependent on the age of the culture. We have selected two types of bacteria, Kocuria rhizophila and Alcagenes faecalis, and we have monitored in equal time intervals of their development the autofluorescence spectra. The stages of development were monitored separately by measuring the turbidity and creating a growth curve. The goal of this study is to find out if the previously observed difference in the autofluorescence spectra of Gram-positive and Gram-negative bacteria is dependent on the stage of the development of the bacterial culture.

Blust, Brittni

2012-02-01

199

Predominant Gram-Positive Bacteria in Human Feces: Numbers, Variety, and Persistence  

PubMed Central

The predominant gram-positive bacteria in 47 fecal specimens from 10 healthy men were studied by microscopic and cultural counts, by the characterization and tentative identification of isolates, and by the use of fluorescein isothiocyanate (FITC)-conjugated globulins prepared using some of the isolates. Gram-positive bacteria averaged 1010.5±0.4(sd/g (wet weight) of feces with significant variation from host to host. Characterization of 865 isolates, all strict anaerobes and carbohydrate fermenters, showed 12 to 39 distinguishable strains from each host and indicated that some strains were present the full period of about 18 months. Sixty percent of the isolates belonged to one of five types, tentatively identified with five species—Bifidobacterium adolescentis, Eubacterium aerofaciens, E. rectale, Peptostreptococcus productus, and Ruminococcus bromii. There was distinct host idiosyncrasy in the pattern of estimated counts of these five types. Certain strains resembling B. adolescentis, E. aerofaciens, and P. productus, distinguished with FITC conjugates, were resident in their hosts for many months. In direct smears each strain constituted about 1% of the total bacteria.

Gossling, Jennifer; Slack, John M.

1974-01-01

200

Surface-conjugated antimicrobial peptide leucocin a displays high binding to pathogenic gram-positive bacteria.  

PubMed

Leucocin A, a representative class IIa bacteriocin, is a ribosomally synthesized antimicrobial peptide (AMP) that displays potent activity against specific gram-positive bacteria. The antibacterial activity of such peptides is preceded by the binding event that can be utilized for studying specific peptide-bacteria interactions. In this study, 37-residue Leucocin A (LeuA) was synthesized using solid-phase peptide synthesis and covalently immobilized on gold substrates from either the N- or C-terminal. Both the peptide monolayers on gold substrates were incubated separately with five strains of gram-positive bacteria and displayed differential binding to different strains with highest binding to pathogenic Listeria monocytogenes . The C-terminally immobilized LeuA showed higher bacterial binding compared to the N-terminally attached LeuA. The full length immobilized LeuA (37-residue) was active as well as displayed higher bacterial binding (73 ± 6 bacteria/100 ?m(2)) compared to 24-residue inactive LeuA fragment (40 ± 8 bacteria/100 ?m(2)) from the C-terminal region. The high and specific bacterial binding ability of LeuA functionalized surfaces support the potential use of class IIa bacteriocins in antimicrobial peptide-based diagnostic platforms. PMID:24359454

Etayash, Hashem; Norman, Lana; Thundat, Thomas; Stiles, Michael; Kaur, Kamaljit

2014-01-22

201

NClassG+: A classifier for non-classically secreted Gram-positive bacterial proteins  

PubMed Central

Background Most predictive methods currently available for the identification of protein secretion mechanisms have focused on classically secreted proteins. In fact, only two methods have been reported for predicting non-classically secreted proteins of Gram-positive bacteria. This study describes the implementation of a sequence-based classifier, denoted as NClassG+, for identifying non-classically secreted Gram-positive bacterial proteins. Results Several feature-based classifiers were trained using different sequence transformation vectors (frequencies, dipeptides, physicochemical factors and PSSM) and Support Vector Machines (SVMs) with Linear, Polynomial and Gaussian kernel functions. Nested k-fold cross-validation (CV) was applied to select the best models, using the inner CV loop to tune the model parameters and the outer CV group to compute the error. The parameters and Kernel functions and the combinations between all possible feature vectors were optimized using grid search. Conclusions The final model was tested against an independent set not previously seen by the model, obtaining better predictive performance compared to SecretomeP V2.0 and SecretPV2.0 for the identification of non-classically secreted proteins. NClassG+ is freely available on the web at http://www.biolisi.unal.edu.co/web-servers/nclassgpositive/

2011-01-01

202

Bioengineered Nisin A Derivatives with Enhanced Activity against Both Gram Positive and Gram Negative Pathogens  

PubMed Central

Nisin is a bacteriocin widely utilized in more than 50 countries as a safe and natural antibacterial food preservative. It is the most extensively studied bacteriocin, having undergone decades of bioengineering with a view to improving function and physicochemical properties. The discovery of novel nisin variants with enhanced activity against clinical and foodborne pathogens has recently been described. We screened a randomized bank of nisin A producers and identified a variant with a serine to glycine change at position 29 (S29G), with enhanced efficacy against S. aureus SA113. Using a site-saturation mutagenesis approach we generated three more derivatives (S29A, S29D and S29E) with enhanced activity against a range of Gram positive drug resistant clinical, veterinary and food pathogens. In addition, a number of the nisin S29 derivatives displayed superior antimicrobial activity to nisin A when assessed against a range of Gram negative food-associated pathogens, including E. coli, Salmonella enterica serovar Typhimurium and Cronobacter sakazakii. This is the first report of derivatives of nisin, or indeed any lantibiotic, with enhanced antimicrobial activity against both Gram positive and Gram negative bacteria.

Field, Des; Begley, Maire; O'Connor, Paula M.; Daly, Karen M.; Hugenholtz, Floor; Cotter, Paul D.; Hill, Colin; Ross, R. Paul

2012-01-01

203

Acylation of SC4 dodecapeptide increases bactericidal potency against Gram-positive bacteria, including drug-resistant strains.  

PubMed Central

We have conjugated dodecyl and octadecyl fatty acids to the N-terminus of SC4, a potently bactericidal, helix-forming peptide 12-mer (KLFKRHLKWKII), and examined the bactericidal activities of the resultant SC4 'peptide-amphiphile' molecules. SC4 peptide-amphiphiles showed up to a 30-fold increase in bactericidal activity against Gram-positive strains (Staphylococcus aureus, Streptococcus pyogenes and Bacillus anthracis), including S. aureus strains resistant to conventional antibiotics, but little or no increase in bactericidal activity against Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa). Fatty acid conjugation improved endotoxin (lipopolysaccharide) neutralization by 3- to 6-fold. Although acylation somewhat increased lysis of human erythrocytes, it did not increase lysis of endothelial cells, and the haemolytic effects occurred at concentrations 10- to 100-fold higher than those required for bacterial cell lysis. For insight into the mechanism of action of SC4 peptide-amphiphiles, CD, NMR and fluorescence spectroscopy studies were performed in micelle and liposome models of eukaryotic and bacterial cell membranes. CD indicated that SC4 peptide-amphiphiles had the strongest helical tendencies in liposomes mimicking bacterial membranes, and strong membrane integration of the SC4 peptide-amphiphiles was observed using tryptophan fluorescence spectroscopy under these conditions; results that correlated with the increased bactericidal activities of SC4 peptide-amphiphiles. NMR structural analysis in micelles demonstrated that the two-thirds of the peptide closest to the fatty acid tail exhibited a helical conformation, with the positively-charged side of the amphipathic helix interacting more with the model membrane surface. These results indicate that conjugation of a fatty acid chain to the SC4 peptide enhances membrane interactions, stabilizes helical structure in the membrane-bound state and increases bactericidal potency.

Lockwood, Nathan A; Haseman, Judith R; Tirrell, Matthew V; Mayo, Kevin H

2004-01-01

204

Isolation and characterization of the solvent-tolerant Bacillus cereus strain R1.  

PubMed

Toluene-tolerant gram-positive bacteria were isolated and identified to belong to the genus Bacillus. They grew in a medium containing yeast extract and in the presence of a separate phase of toluene or other hydrocarbons, but not when aliphatic alcohols were present. The isolate Bacillus cereus R1 did not metabolise or transform toluene. Toluene accumulation in its cells was rapid, unless the organism was supplied with glucose as energy source. In bacteria adapted to toluene, the amount of toluene accumulating in cells was one-half that in nonadapted bacteria. Valinomycin (K+ ionophore) and o-vanadate (ATPase inhibitor) as inhibitors of energy metabolism partly counteracted the effect of glucose as energy source. These results suggest the presence of an efflux mechanism for toluene in strain R1. The nature of this mechanism and its function in a solvent-tolerant gram-positive strain are discussed. PMID:16233268

Matsumoto, Michiaki; de Bont, Jan A M; Isken, Sonja

2002-01-01

205

The potent antimicrobial properties of cell penetrating peptide-conjugated silver nanoparticles with excellent selectivity for gram-positive bacteria over erythrocytes.  

PubMed

Silver nanoparticles are of great interest for use as antimicrobial agents. Studies aimed at producing potent nano-silver biocides have focused on manipulation of particle size, shape, composition and surface charge. Here, we report the cell penetrating peptide catalyzed formation of antimicrobial silver nanoparticles in N,N-dimethylformamide. The novel nano-composite demonstrated a distinctly enhanced biocidal effect toward bacteria (gram-positive Bacillus subtilis, gram-negative Escherichia coli) and pathogenic yeast (Candida albicans), as compared to triangular and extremely small silver nanoparticles. In addition, a satisfactory biocompatibility was verified by a haemolysis test. Our results provide a paradigm in developing strategies that can maximize the silver nanoparticle application potentials while minimizing the toxic effects. PMID:23525222

Liu, Lihong; Yang, Jun; Xie, Jianping; Luo, Zhentao; Jiang, Jiang; Yang, Yi Yan; Liu, Shaomin

2013-05-01

206

Genomic and Enzymatic Results Show Bacillus cellulosilyticus Uses a Novel Set of LPXTA Carbohydrases to Hydrolyze Polysaccharides  

PubMed Central

Background Alkaliphilic Bacillus species are intrinsically interesting due to the bioenergetic problems posed by growth at high pH and high salt. Three alkaline cellulases have been cloned, sequenced and expressed from Bacillus cellulosilyticus N-4 (Bcell) making it an excellent target for genomic sequencing and mining of biomass-degrading enzymes. Methodology/Principal Findings The genome of Bcell is a single chromosome of 4.7 Mb with no plasmids present and three large phage insertions. The most unusual feature of the genome is the presence of 23 LPXTA membrane anchor proteins; 17 of these are annotated as involved in polysaccharide degradation. These two values are significantly higher than seen in any other Bacillus species. This high number of membrane anchor proteins is seen only in pathogenic Gram-positive organisms such as Listeria monocytogenes or Staphylococcus aureus. Bcell also possesses four sortase D subfamily 4 enzymes that incorporate LPXTA-bearing proteins into the cell wall; three of these are closely related to each other and unique to Bcell. Cell fractionation and enzymatic assay of Bcell cultures show that the majority of polysaccharide degradation is associated with the cell wall LPXTA-enzymes, an unusual feature in Gram-positive aerobes. Genomic analysis and growth studies both strongly argue against Bcell being a truly cellulolytic organism, in spite of its name. Preliminary results suggest that fungal mycelia may be the natural substrate for this organism. Conclusions/Significance Bacillus cellulosilyticus N-4, in spite of its name, does not possess any of the genes necessary for crystalline cellulose degradation, demonstrating the risk of classifying microorganisms without the benefit of genomic analysis. Bcell is the first Gram-positive aerobic organism shown to use predominantly cell-bound, non-cellulosomal enzymes for polysaccharide degradation. The LPXTA-sortase system utilized by Bcell may have applications both in anchoring cellulases and other biomass-degrading enzymes to Bcell itself and in anchoring proteins other Gram-positive organisms.

Mead, David; Drinkwater, Colleen; Brumm, Phillip J.

2013-01-01

207

Identification of pili on the surface of Finegoldia magna - A Gram-positive anaerobic cocci.  

PubMed

Pili have only been discovered in the major Gram-positive pathogens in the past decade and they have been found to play an important role in colonisation and virulence. Pili have been shown to have many important functions including attachment to host tissues, mediating bacterial aggregation, biofilm formation and binding to proteins in the extracellular matrix. In this study, sortase-dependent pili have been found to be expressed on the surface of Finegoldia magna ALB8. F. magna is a Gram-positive anaerobic coccus that, primarily, is a commensal of the skin and mucous membranes, but has also been isolated from various clinical infection sites and is associated with soft-tissue abscesses, wound infections and bone and prosthetic joint infections. In this study, F. magna ALB8 was found to harbour three sortases at the pilus locus, two of which bear high similarity to class C sortases in Streptococcus pneumoniae. Two putative sortase-dependent pili proteins were found in the locus, with one being identified as the major pilus subunit, Fmp1 (F. magna pilus subunit 1), due to its high similarity to other major pilus proteins in prominent Gram-positive pathogens. The presence of sortase-dependent pili was confirmed experimentally through recombinant production of Fmp1 and production of antiserum. The Fmp1 antiserum was used in Western blot to show the presence of a high molecular weight protein ladder, characteristic of the presence of pili, in trypsin released cell wall surface proteins from F. magna. The presence of sortase-dependent pili was visually confirmed by transmission electron microscopy, which showed the binding of gold labelled anti-Fmp1 to individual pilus proteins along the pilus. Furthermore, pili could also be found to bind and interact with keratinocytes in the epidermal layer of human skin, suggesting an adhesive role for pili on F. magna. Our work represents the first description of pilus structures in F. magna. This discovery further elucidates F. magna physiology and allows for additional analysis of host-bacterial interactions in future studies. PMID:24685556

Murphy, Elizabeth C; Janulczyk, Robert; Karlsson, Christofer; Mörgelin, Matthias; Frick, Inga-Maria

2014-06-01

208

Ceftaroline activity tested against uncommonly isolated Gram-positive pathogens: report from the SENTRY Antimicrobial Surveillance Program (2008-2011).  

PubMed

Ceftaroline was tested against 1859 clinically significant Gram-positive organisms from uncommonly isolated species. The organisms (31 species/groups) were collected from 133 medical centres worldwide over a 4-year period (2008-2011). Coagulase-negative staphylococci were generally susceptible to ceftaroline, with MIC50 values (minimum inhibitory concentration required to inhibit 50% of the isolates) of 0.06-0.5mg/L. Ceftaroline was active against Micrococcus spp. [minimum inhibitory concentration required to inhibit 90% of the isolates (MIC90)=0.06 mg/L], but showed more limited potency versus some Corynebacterium spp. and Listeria monocytogenes isolates. Ceftaroline was active against all ?-haemolytic streptococci and viridans group streptococcal species/groups listed, with MIC50 and MIC90 values ranging from ? 0.015 mg/L to 0.03 mg/L and from ? 0.015 mg/L to 0.5mg/L, respectively. Based on these in vitro findings, ceftaroline may have a potential role in the treatment of infections caused by these rarer species as guided by reference MIC test results. PMID:24342717

Sader, Helio S; Jones, Ronald N; Stilwell, Matthew G; Flamm, Robert K

2014-03-01

209

The Clavibacter michiganensis subspecies: molecular investigation of gram-positive bacterial plant pathogens.  

PubMed

Clavibacter michiganensis subspecies are actinomycete plant pathogens residing mainly in the xylem vessels that infect economically important host plants. In the Clavibacter subspecies michiganensis and sepedonicus, infecting tomato and potato, respectively, essential factors for disease induction are plasmid encoded and loss of the virulence plasmids converts these biotrophic pathogens into endophytes. The genes responsible for successful colonization of the host plant, including evasion/suppression of plant defense reactions, are chromosomally encoded. Several serine proteases seem to be involved in colonization. They are secreted by Clavibacter, but their targets remain unknown. A type 3 secretion system (T3SS) translocating effectors into the plant cells is absent in these gram-positive pathogens. With the development of the modern 'omics technologies for RNA and proteins based on the known genome sequences, a new phase in the investigation of the mechanisms of plant pathogenicity has begun to allow the genome-wide investigation of the Clavibacter-host interaction. PMID:21438679

Eichenlaub, Rudolf; Gartemann, Karl-Heinz

2011-01-01

210

Activation and manipulation of host responses by a Gram-positive bacterium.  

PubMed

The interaction between tomato plants and Clavibacter michiganensis subsp. michiganensis (Cmm) represents a model pathosystem to study the interplay between the virulence determinants of a Gram-positive bacterium and the attempt of a crop plant to counteract pathogen invasion. To investigate plant responses activated during this compatible interaction, we recently analyzed gene expression profiles of tomato stems infected with Cmm. This analysis revealed activation of basal defense responses that are typically observed upon plant perception of pathogen-associated molecular patterns. In addition, Cmm infection upregulated the expression of host genes related to ethylene synthesis and response. Further analysis of tomato plants impaired in ethylene perception and production demonstrated an important role for ethylene in the development of disease symptoms. Here we discuss possible molecular strategies used by the plant to recognize Cmm infection and possible mechanisms employed by the pathogen to interfere with the activation of plant defense responses and promote disease. PMID:19704516

Balaji, Vasudevan; Sessa, Guido

2008-10-01

211

Sonodynamic Excitation of Rose Bengal for Eradication of Gram-Positive and Gram-Negative Bacteria  

PubMed Central

Photodynamic antimicrobial chemotherapy based on photosensitizers activated by illumination is limited by poor penetration of visible light through skin and tissues. In order to overcome this problem, Rose Bengal was excited in the dark by 28?kHz ultrasound and was applied for inactivation of bacteria. It is demonstrated, for the first time, that the sonodynamic technique is effective for eradication of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli. The net sonodynamic effect was calculated as a 3-4 log10 reduction in bacteria concentration, depending on the cell and the Rose Bengal concentration and the treatment time. Sonodynamic treatment may become a novel and effective form of antimicrobial therapy and can be used for low-temperature sterilization of medical instruments and surgical accessories.

Nakonechny, Faina; Nisnevitch, Michael; Nitzan, Yeshayahu; Nisnevitch, Marina

2013-01-01

212

Di-alkylated paromomycin derivatives: targeting the membranes of gram positive pathogens that cause skin infections.  

PubMed

A collection of paromomycin-based di-alkylated cationic amphiphiles differing in the lengths of their aliphatic chain residues were designed, synthesized, and evaluated against 14 Gram positive pathogens that are known to cause skin infections. Paromomycin derivatives that were di-alkylated with C7 and C8 linear aliphatic chains had improved antimicrobial activities relative to the parent aminoglycoside as well as to the clinically used membrane-targeting antibiotic gramicidin D; several novel derivatives were at least 16-fold more potent than the parent aminoglycoside paromomycin. Comparison between a di-alkylated and a mono-alkylated paromomycin indicated that the di-alkylation strategy leads to both an improvement in antimicrobial activity and to a dramatic reduction in undesired red blood cell hemolysis caused by many aminoglycoside-based cationic amphiphiles. Scanning electron microscopy provided evidence for cell surface damage by the reported di-alkylated paromomycins. PMID:23602621

Berkov-Zrihen, Yifat; Herzog, Ido M; Feldman, Mark; Sonn-Segev, Adar; Roichman, Yael; Fridman, Micha

2013-06-15

213

The immune response after stimulation with wall components of gram-positive bacteria and fungi.  

PubMed

Although several components of the microbial wall of gram-positive bacteria and fungi possess immunostimulatory properties, their pathogenetic role remains incompletely evaluated. The purpose of this study was to assess the basic immune status of patients susceptible to infections and their capability for cytokine production after stimulation with wall components of gram-positive bacteria and fungi. We measured serum cytokine levels as well as cytokine production after ex vivo lipoteichoic acid (LTA) and mannan stimulation of whole blood. The blood was taken from 10 healthy volunteers, 10 patients with end-stage renal disease (ESRD), 10 patients with diabetes mellitus (DM), and 10 patients on their 2nd day of stay in the Intensive Care Unit (ICU), who suffered from non septic systemic inflammatory response syndrome (SIRS) and had an APACHE II score ?25. We used 1 ?g/ml LTA and 100 ?g/ml mannan for an incubation period of 8 h to stimulate 100 ?l aliquots of whole blood. All patient groups had higher baseline values of TNF-?, IL-6, IL-1?, and IL-10 compared to the control group, but only for ICU patients the difference was statistically significant. The ratio IL-10/IL-6 was found 0.33, 0.22, and 0.96 in healthy persons, ESRD, and DM patients respectively, and 1.32 in ICU patients. In all examined groups, the levels of cytokines significantly increased after stimulation by LTA and mannan, although in severely ill patients this change was considerably smaller, possibly reflecting a state of monocytes' depression and relative hyporesponsiveness. No significant differences between the LTA and the mannan stimulation were observed. PMID:24440200

Tsigou, Evdoxia; Stavros, Aloizos; Pavlos, Myrianthefs; Stavros, Gourgiotis; Athanassios, Tsakris; George, Baltopoulos

2014-01-01

214

sRNAdb: A small non-coding RNA database for gram-positive bacteria  

PubMed Central

Background The class of small non-coding RNA molecules (sRNA) regulates gene expression by different mechanisms and enables bacteria to mount a physiological response due to adaptation to the environment or infection. Over the last decades the number of sRNAs has been increasing rapidly. Several databases like Rfam or fRNAdb were extended to include sRNAs as a class of its own. Furthermore new specialized databases like sRNAMap (gram-negative bacteria only) and sRNATarBase (target prediction) were established. To the best of the authors’ knowledge no database focusing on sRNAs from gram-positive bacteria is publicly available so far. Description In order to understand sRNA’s functional and phylogenetic relationships we have developed sRNAdb and provide tools for data analysis and visualization. The data compiled in our database is assembled from experiments as well as from bioinformatics analyses. The software enables comparison and visualization of gene loci surrounding the sRNAs of interest. To accomplish this, we use a client–server based approach. Offline versions of the database including analyses and visualization tools can easily be installed locally on the user’s computer. This feature facilitates customized local addition of unpublished sRNA candidates and related information such as promoters or terminators using tab-delimited files. Conclusion sRNAdb allows a user-friendly and comprehensive comparative analysis of sRNAs from available sequenced gram-positive prokaryotic replicons. Offline versions including analysis and visualization tools facilitate complex user specific bioinformatics analyses.

2012-01-01

215

Atopococcus tabaci gen. nov., sp. nov., a novel Gram-positive, catalase-negative, coccus-shaped bacterium isolated from tobacco.  

PubMed

A novel Gram-positive, aerobic, catalase-negative, coccus-shaped organism originating from tobacco was characterized using phenotypic and molecular taxonomic methods. The organism contained a cell wall murein based on L-lysine (variation A4alpha, type L-lysine-L-glutamic acid), synthesized long-chain cellular fatty acids of the straight-chain saturated and monounsaturated types (with C(16:1)omega9, C(16:0) and C(18:1)omega9 predominating) and possessed a DNA G+C content of 46 mol%. Based on morphological, biochemical and chemical characteristics, the coccus-shaped organism did not conform to any presently recognized taxon. Comparative 16S rRNA gene sequencing studies confirmed the distinctiveness of the unknown coccus, with the bacterium displaying sequence divergence values of greater than 7% with other recognized Gram-positive taxa. Treeing analysis reinforced its distinctiveness, with the unidentified organism forming a relatively long subline branching at the periphery of an rRNA gene sequence cluster which encompasses the genera Alloiococcus, Allofustis, Alkalibacterium, Atopostipes, Dolosigranulum and Marinilactibacillus. Based on phenotypic and molecular phylogenetic evidence, it is proposed that the unknown organism from tobacco be classified as a new genus and species, Atopococcus tabaci gen. nov., sp. nov. The type strain of Atopococcus tabaci is CCUG 48253(T) (=CIP 108502(T)). PMID:16014503

Collins, Matthew D; Wiernik, Anna; Falsen, Enevold; Lawson, Paul A

2005-07-01

216

Identification and characterization of a new xylanase from Gram-positive bacteria isolated from termite gut (Reticulitermes santonensis).  

PubMed

Termites are world champions at digesting lignocellulosic compounds, thanks to cooperation between their own enzymes and exogenous enzymes from microorganisms. Prokaryotic cells are responsible for a large part of this lignocellulolytic activity. Bacterial enzyme activities have been demonstrated in the higher and the lower termite gut. From five clones of Gram-positive bacteria isolated and identified in a previous work, we constructed a genomic DNA library and performed functional screening for alpha-amylase, beta-glucosidase, and xylanase activities. One candidate, Xyl8B8, showed xylanase activity. Sequence analysis of the genomic insert revealed five complete ORFs on the cloned DNA (5746bp). Among the encoded proteins were a putative endo-1,4-beta-xylanase (XylB8) belonging to glycoside hydrolase family 11 (GH11). On the basis of sequence analyses, genomic DNA organization, and phylogenetic analysis, the insert was shown to come from an actinobacterium. The mature xylanase (mXylB8) was expressed in Escherichia coli and purified by affinity chromatography and detected by zymogram analysis after renaturing. It showed maximal xylanase activity in sodium acetate buffer, pH 5.0 at 55 °C. Its activity was increased by reducing agents and decreased by Cu(2+), some detergents, and chelating agents. Its substrate specificity appeared limited to xylan. PMID:22487213

Mattéotti, Christel; Bauwens, Julien; Brasseur, Catherine; Tarayre, Cédric; Thonart, Philippe; Destain, Jacqueline; Francis, Frédéric; Haubruge, Eric; De Pauw, Edwin; Portetelle, Daniel; Vandenbol, Micheline

2012-06-01

217

DNA Repair and Genome Maintenance in Bacillus subtilis  

PubMed Central

Summary: From microbes to multicellular eukaryotic organisms, all cells contain pathways responsible for genome maintenance. DNA replication allows for the faithful duplication of the genome, whereas DNA repair pathways preserve DNA integrity in response to damage originating from endogenous and exogenous sources. The basic pathways important for DNA replication and repair are often conserved throughout biology. In bacteria, high-fidelity repair is balanced with low-fidelity repair and mutagenesis. Such a balance is important for maintaining viability while providing an opportunity for the advantageous selection of mutations when faced with a changing environment. Over the last decade, studies of DNA repair pathways in bacteria have demonstrated considerable differences between Gram-positive and Gram-negative organisms. Here we review and discuss the DNA repair, genome maintenance, and DNA damage checkpoint pathways of the Gram-positive bacterium Bacillus subtilis. We present their molecular mechanisms and compare the functions and regulation of several pathways with known information on other organisms. We also discuss DNA repair during different growth phases and the developmental program of sporulation. In summary, we present a review of the function, regulation, and molecular mechanisms of DNA repair and mutagenesis in Gram-positive bacteria, with a strong emphasis on B. subtilis.

Lenhart, Justin S.; Schroeder, Jeremy W.; Walsh, Brian W.

2012-01-01

218

Fructose Utilization in Lactococcus lactis as a Model for Low-GC Gram-Positive Bacteria: Its Regulator, Signal, and DNA-Binding Site  

PubMed Central

In addition to its role as carbon and energy source, fructose metabolism was reported to affect other cellular processes, such as biofilm formation by streptococci and bacterial pathogenicity in plants. Fructose genes encoding a 1-phosphofructokinase and a phosphotransferase system (PTS) fructose-specific enzyme IIABC component reside commonly in a gene cluster with a DeoR family regulator in various gram-positive bacteria. We present a comprehensive study of fructose metabolism in Lactococcus lactis, including a systematic study of fru mutants, global messenger analysis, and a molecular characterization of its regulation. The fru operon is regulated at the transcriptional level by both FruR and CcpA and at the metabolic level by inducer exclusion. The FruR effector is fructose-1-phosphate (F1P), as shown by combined analysis of transcription and measurements of the intracellular F1P pools in mutants either unable to produce this metabolite or accumulating it. The regulation of the fru operon by FruR requires four adjacent 10-bp direct repeats. The well-conserved organization of the fru promoter region in various low-GC gram-positive bacteria, including CRE boxes as well as the newly defined FruR motif, suggests that the regulation scheme defined in L. lactis could be applied to these bacteria. Transcriptome profiling of fruR and fruC mutants revealed that the effect of F1P and FruR regulation is limited to the fru operon in L. lactis. This result is enforced by the fact that no other targets for FruR were found in the available low-GC gram-positive bacteria genomes, suggesting that additional phenotypical effects due to fructose metabolism do not rely directly on FruR control, but rather on metabolism.

Barriere, Charlotte; Veiga-da-Cunha, Maria; Pons, Nicolas; Guedon, Eric; van Hijum, Sacha A. F. T.; Kok, Jan; Kuipers, Oscar P.; Ehrlich, Dusko S.; Renault, Pierre

2005-01-01

219

Bacillus algicola sp. nov., a novel filamentous organism isolated from brown alga Fucus evanescens.  

PubMed

A slightly yellowish, Gram-positive, filamentous with 'cross-like' branching, aerobic, spore-forming bacterium was isolated from enrichment culture during degradation of the thallus of the brown alga Fucus evanescens. The bacterium studied was chemoorganotrophic, tolerant to 3% NaCl, alkalitolerant, and alginolytic. The predominant cellular fatty acid was ai15:0 which accounted more than 65% of total fatty acids, while i14:0, il5:0 i16:0, and ai17:0 made up 25%. DNA base composition was 37 mol% GC. Phylogenetic analysis of 16S rDNA gene revealed that this isolate was a member of the genus Bacillus, with no close relatives at the species level (16S rRNA gene sequence similarity less 97%). On the basis of the significant differences demonstrated in the phenotypic and chemotaxonomic characteristics, it is suggested that the bacterium be classified as a novel species; the name Bacillus algicola sp. nov. is proposed. The type strain is KMM 3737T (= CIP 107850T). PMID:15214635

Ivanova, Elena P; Alexeeva, Yulia A; Zhukova, Natalia V; Gorshkova, Natalia M; Buljan, Vlado; Nicolau, Dan V; Mikhailov, Valery V; Christen, Richard

2004-05-01

220

Global mRNA decay analysis at single nucleotide resolution reveals segmental and positional degradation patterns in a Gram-positive bacterium  

PubMed Central

Background Recent years have shown a marked increase in the use of next-generation sequencing technologies for quantification of gene expression (RNA sequencing, RNA-Seq). The expression level of a gene is a function of both its rate of transcription and RNA decay, and the influence of mRNA decay rates on gene expression in genome-wide studies of Gram-positive bacteria is under-investigated. Results In this work, we employed RNA-Seq in a genome-wide determination of mRNA half-lives in the Gram-positive bacterium Bacillus cereus. By utilizing a newly developed normalization protocol, RNA-Seq was used successfully to determine global mRNA decay rates at the single nucleotide level. The analysis revealed positional degradation patterns, with mRNAs being degraded from both ends of the molecule, indicating that both 5' to 3' and 3' to 5' directions of RNA decay are present in B. cereus. Other operons showed segmental degradation patterns where specific ORFs within polycistrons were degraded at variable rates, underlining the importance of RNA processing in gene regulation. We determined the half-lives for more than 2,700 ORFs in B. cereus ATCC 10987, ranging from less than one minute to more than fifteen minutes, and showed that mRNA decay rate correlates globally with mRNA expression level, GC content, and functional class of the ORF. Conclusions To our knowledge, this study presents the first global analysis of mRNA decay in a bacterium at single nucleotide resolution. We provide a proof of principle for using RNA-Seq in bacterial mRNA decay analysis, revealing RNA processing patterns at the single nucleotide level.

2012-01-01

221

Revised mechanism of D-alanine incorporation into cell wall polymers in Gram-positive bacteria.  

PubMed

Teichoic acids (TAs) are important for growth, biofilm formation, adhesion and virulence of Gram-positive bacterial pathogens. The chemical structures of the TAs vary between bacteria, though they typically consist of zwitterionic polymers that are anchored to either the peptidoglycan layer as in the case of wall teichoic acid (WTA) or the cell membrane and named lipoteichoic acid (LTA). The polymers are modified with D-alanines and a lack of this decoration leads to increased susceptibility to cationic antimicrobial peptides. Four proteins, DltA-D, are essential for the incorporation of d-alanines into cell wall polymers and it has been established that DltA transfers D-alanines in the cytoplasm of the cell onto the carrier protein DltC. However, two conflicting models have been proposed for the remainder of the mechanism. Using a cellular protein localization and membrane topology analysis, we show here that DltC does not traverse the membrane and that DltD is anchored to the outside of the cell. These data are in agreement with the originally proposed model for D-alanine incorporation through a process that has been proposed to proceed via a D-alanine undecaprenyl phosphate membrane intermediate. Furthermore, we found that WTA isolated from a Staphylococcus aureus strain lacking LTA contains only a small amount of D-alanine, indicating that LTA has a role, either direct or indirect, in the efficient D-alanine incorporation into WTA in living cells. PMID:23858088

Reichmann, Nathalie T; Cassona, Carolina Picarra; Gründling, Angelika

2013-09-01

222

Population biology of Gram-positive pathogens: high-risk clones for dissemination of antibiotic resistance  

PubMed Central

Infections caused by multi-resistant Gram positive bacteria represent a major health burden in the community as well as in hospitalized patients. Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium are well-known pathogens of hospitalized patients, frequently linked with resistance against multiple antibiotics, compromising effective therapy. Streptococcus pneumoniae and Streptococcus pyogenes are important pathogens in the community and S. aureus has recently emerged as an important community-acquired pathogen. Population genetic studies reveal that recombination prevails as a driving force of genetic diversity in E. faecium, E. faecalis, S. pneumoniae, and S. pyogenes and thus, these species are weakly clonal. Although recombination has a relatively modest role driving the genetic variation of the core genome of S. aureus, the horizontal acquistion of resistance and virulence genes plays a key role in the emergence of new clinically relevant clones in this species. In this review we discuss the population genetics of E. faecium, E. faecalis, S. pneumoniae, S. pyogenes, and S. aureus. Knowledge of the population structure of these pathogens is not only highly relevant for (molecular) epidemiological research but also for identifying the genetic variation that underlies changes in clinical behaviour, to improve our understanding of the pathogenic behaviour of particular clones and to identify novel targets for vaccines or immunotherapy.

Willems, Rob J. L.; Hanage, William P; Bessen, Debra E.; Feil, Edward J.

2011-01-01

223

Mechanistic antimicrobial approach of extracellularly synthesized silver nanoparticles against gram positive and gram negative bacteria.  

PubMed

The development of eco-friendly and reliable processes for the synthesis of nanoparticles has attracted considerable interest in nanotechnology. In this study, an extracellular enzyme system of a newly isolated microorganism, Exiguobacterium sp. KNU1, was used for the reduction of AgNO? solutions to silver nanoparticles (AgNPs). The extracellularly biosynthesized AgNPs were characterized by UV-vis spectroscopy, Fourier transform infra-red spectroscopy and transmission electron microscopy. The AgNPs were approximately 30 nm (range 5-50 nm) in size, well-dispersed and spherical. The AgNPs were evaluated for their antimicrobial effects on different gram negative and gram positive bacteria using the minimum inhibitory concentration method. Reasonable antimicrobial activity against Salmonella typhimurium, Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus was observed. The morphological changes occurred in all the microorganisms tested. In particular, E. coli exhibited DNA fragmentation after being treated with the AgNPs. Finally, the mechanism for their bactericidal activity was proposed according to the results of scanning electron microscopy and single cell gel electrophoresis. PMID:23867968

Tamboli, Dhawal P; Lee, Dae Sung

2013-09-15

224

Mannopeptimycins, a novel class of glycopeptide antibiotics active against gram-positive bacteria.  

PubMed

Mannopeptimycins alpha-epsilon, novel glycopeptides with activity against methicillin-resistant staphylococci and vancomycin-resistant enterococci, are purified from the fermentation broth of a strain of Streptomyces hygroscopicus, LL-AC98, and their structures characterized using spectroscopic analyses and chemical methods. The SAR data of the natural and synthetic esters demonstrate that the presence of hydrophobic groups near the terminal mannosyl moiety is critical for antibacterial potency. Scalable syntheses of 4,6-cyclic acetals and ketals on this moiety are used to produce significant quantities of the respective mannopeptimycin derivatives. These acetal and ketal derivatives exhibit potent activities against susceptible and resistant Gram-positive bacteria in both in vitro and in vivo experiments, comparable with or exceeding the activity of vancomycin. Studies on the mechanism of action suggest that the mannopeptimycins interfere with the late stages of bacterial cell wall biosynthesis. It is believed that these antibiotics inhibit the transglycosylation by binding to the transglycosylase substrate, lipid II. PMID:15702316

He, Haiyin

2005-06-01

225

Combination of Pantothenamides with Vanin Inhibitors as a Novel Antibiotic Strategy against Gram-Positive Bacteria  

PubMed Central

The emergence of resistance against current antibiotics calls for the development of new compounds to treat infectious diseases. Synthetic pantothenamides are pantothenate analogs that possess broad-spectrum antibacterial activity in vitro in minimal media. Pantothenamides were shown to be substrates of the bacterial coenzyme A (CoA) biosynthetic pathway, causing cellular CoA depletion and interference with fatty acid synthesis. In spite of their potential use and selectivity for bacterial metabolic routes, these compounds have never made it to the clinic. In the present study, we show that pantothenamides are not active as antibiotics in the presence of serum, and we found that they were hydrolyzed by ubiquitous pantetheinases of the vanin family. To address this further, we synthesized a series of pantetheinase inhibitors based on a pantothenate scaffold that inhibited serum pantetheinase activity in the nanomolar range. Mass spectrometric analysis showed that addition of these pantetheinase inhibitors prevented hydrolysis of pantothenamides by serum. We found that combinations of these novel pantetheinase inhibitors and prototypic pantothenamides like N5-Pan and N7-Pan exerted antimicrobial activity in vitro, particularly against Gram-positive bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, and Streptococcus pyogenes) even in the presence of serum. These results indicate that pantothenamides, when protected against degradation by host pantetheinases, are potentially useful antimicrobial agents.

Jansen, Patrick A. M.; Hermkens, Pedro H. H.; Zeeuwen, Patrick L. J. M.; Botman, Peter N. M.; Blaauw, Richard H.; Burghout, Peter; van Galen, Peter M.; Mouton, Johan W.; Rutjes, Floris P. J. T.

2013-01-01

226

Characterization of unusual alkaliphilic gram-positive bacteria isolated from degraded brown alga thalluses.  

PubMed

Two orange-pigmented Gram-positive, aerobic bacteria were isolated from enrichment culture during degradation of brown alga Fucus evanescens thalluses. In this work, atomic force microscopy (AFM) has been used to study the cell morphology. The non-contact mode imaging revealed unusual irregular coccoid shape of cells, possessing a single flagellum. Bacteria produced carotenoid pigments, were chemo-organotrophic, alkaliphilic and halo-tolerant growing well on nutrient media containing up to 15% NaCl. Growth temperature ranged from 5 to 45 degrees C. The DNA base compositions were 48 mol% G + C and the level of DNA similarity of two strains was conspecific (98%). A comparative phylogenetic analysis of 16S rRNA gene sequences revealed that the strain KMM 3738 tightly clustered with recently described Planococcus maritimus (99.9% 16S rRNA gene sequence similarity). DNA-DNA hybridisation experiments revealed that DNA from the KMM 3738 showed 12-15% and 16-35% of genetic relatedness with the DNA of type strains of the genera Planomicrobium and Planococcus, respectively, and 87% with DNA from Planococcus maritimus, indicating that new isolates belong to the later species. PMID:17100323

Ivanova, E P; Wright, J P; Lysenko, A M; Zhukova, N V; Alexeeva, Y V; Buljan, V; Kalinovskaya, N I; Nicolau, D V; Christen, R; Mikhailov, V V

2006-01-01

227

Population biology of Gram-positive pathogens: high-risk clones for dissemination of antibiotic resistance.  

PubMed

Infections caused by multiresistant Gram-positive bacteria represent a major health burden in the community as well as in hospitalized patients. Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium are well-known pathogens of hospitalized patients, frequently linked with resistance against multiple antibiotics, compromising effective therapy. Streptococcus pneumoniae and Streptococcus pyogenes are important pathogens in the community and S. aureus has recently emerged as an important community-acquired pathogen. Population genetic studies reveal that recombination prevails as a driving force of genetic diversity in E. faecium, E. faecalis, S. pneumoniae and S. pyogenes, and thus, these species are weakly clonal. Although recombination has a relatively modest role driving the genetic variation of the core genome of S. aureus, the horizontal acquisition of resistance and virulence genes plays a key role in the emergence of new clinically relevant clones in this species. In this review, we discuss the population genetics of E. faecium, E. faecalis, S. pneumoniae, S. pyogenes and S. aureus. Knowledge of the population structure of these pathogens is not only highly relevant for (molecular) epidemiological research but also for identifying the genetic variation that underlies changes in clinical behaviour, to improve our understanding of the pathogenic behaviour of particular clones and to identify novel targets for vaccines or immunotherapy. PMID:21658083

Willems, Rob J L; Hanage, William P; Bessen, Debra E; Feil, Edward J

2011-09-01

228

Gene Targeting in the Gram-Positive Bacterium Lactococcus lactis, Using Various Delta Ribozymes  

PubMed Central

The trans-acting antigenomic delta ribozyme, isolated from the human hepatitis delta virus, was shown to be highly stable and active in vitro, as well as in mammalian cell lines. However, the stability and gene-targeting competence of this small ribozyme have not been studied previously in bacterial cells. In this paper we describe the use of two variants of the trans-acting antigenomic delta ribozyme targeting the abundant EF-Tu mRNA in the industrially important gram-positive bacterium Lactococcus lactis. These two delta ribozyme variants were expressed at significant levels and were shown to be highly stable in vivo. The half-life of the EF-Tu mRNA was slightly but consistently reduced in the presence of the classical delta ribozymes (7 to 13%). In contrast, delta ribozymes harboring a specific on/off riboswitch (SOFA-delta ribozymes) targeting the same sites on the EF-Tu mRNA considerably reduced the half-life of this mRNA (22 to 47%). The rates of catalysis of the SOFA-delta ribozymes in L. lactis were similar to the rates determined in vitro, showing that this new generation of delta ribozymes was highly efficient in these bacterial cells. Clearly, SOFA-delta ribozymes appear to be an ideal means for development of gene inactivation systems in bacteria.

Fiola, Karine; Perreault, Jean-Pierre; Cousineau, Benoit

2006-01-01

229

Sortases and the Art of Anchoring Proteins to the Envelopes of Gram-Positive Bacteria  

PubMed Central

The cell wall envelopes of gram-positive bacteria represent a surface organelle that not only functions as a cytoskeletal element but also promotes interactions between bacteria and their environment. Cell wall peptidoglycan is covalently and noncovalently decorated with teichoic acids, polysaccharides, and proteins. The sum of these molecular decorations provides bacterial envelopes with species- and strain-specific properties that are ultimately responsible for bacterial virulence, interactions with host immune systems, and the development of disease symptoms or successful outcomes of infections. Surface proteins typically carry two topogenic sequences, i.e., N-terminal signal peptides and C-terminal sorting signals. Sortases catalyze a transpeptidation reaction by first cleaving a surface protein substrate at the cell wall sorting signal. The resulting acyl enzyme intermediates between sortases and their substrates are then resolved by the nucleophilic attack of amino groups, typically provided by the cell wall cross bridges of peptidoglycan precursors. The surface protein linked to peptidoglycan is then incorporated into the envelope and displayed on the microbial surface. This review focuses on the mechanisms of surface protein anchoring to the cell wall envelope by sortases and the role that these enzymes play in bacterial physiology and pathogenesis.

Marraffini, Luciano A.; DeDent, Andrea C.; Schneewind, Olaf

2006-01-01

230

Copper Acquisition Is Mediated by YcnJ and Regulated by YcnK and CsoR in Bacillus subtilis  

Microsoft Academic Search

Copper is an essential cofactor for many enzymes, and at over a threshold level, it is toxic for all organisms. To understand the mechanisms underlying copper homeostasis of the gram-positive bacterium Bacillus subtilis, we have performed microarray studies under copper-limiting conditions. These studies revealed that the ycnJ gene encodes a protein that plays an important role in copper metabolism, as

Shashi Chillappagari; Marcus Miethke; Hein Trip; Oscar P. Kuipers; Mohamed A. Marahiel

2009-01-01

231

Salirhabdus euzebyi gen. nov., sp. nov., a Gram-positive, halotolerant bacterium isolated from a sea salt evaporation pond.  

PubMed

A low-G+C, Gram-positive bacterium, designated CVS-14(T), was recovered from a sea salt evaporation pond on the island of Sal in the Cape Verde Archipelago. This organism was catalase- and oxidase-positive. Cells were motile, spore-forming aerobic rods, with an optimum growth temperature of about 35-40 degrees C and optimum pH between 7.0 and 8.5. Optimal growth occurred in media containing 4-6 % (w/v) NaCl, although the organism was able to grow in medium without added NaCl and in medium containing 16 % NaCl. The cell-wall peptidoglycan was of A1 gamma type and the major respiratory quinone was menaquinone 7 (MK-7). Major fatty acids were iso-15 : 0, anteiso-15 : 0, iso-17 : 0 and anteiso-17 : 0. The DNA G+C content was 37.0 mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain CVS-14(T) formed a distinct new branch within the radiation of the moderately halophilic bacilli group, forming a separate lineage from species of the genera Salinibacillus, Paucisalibacillus, Oceanobacillus, Lentibacillus and Virgibacillus. Strain CVS-14(T) showed 16S rRNA gene pairwise similarity values of approximately 95 % with species of the genus Salinibacillus. On the basis of morphological, physiological, chemotaxonomic and phylogenetic characteristics, strain CVS-14(T) is considered to represent a novel species in a new genus, for which the name Salirhabdus euzebyi gen. nov., sp. nov. is proposed. The type strain is CVS-14(T) (=LMG 22839(T)=CIP 108577(T)). PMID:17625195

Albuquerque, Luciana; Tiago, Igor; Rainey, Fred A; Taborda, Marco; Nobre, M Fernanda; Veríssimo, António; da Costa, Milton S

2007-07-01

232

Cellular fatty acid composition as an adjunct to the identification of asporogenous, aerobic gram-positive rods.  

PubMed Central

Cellular fatty acid (CFA) compositions of 561 asporogenous, aerobic gram-positive rods were analyzed by gas-liquid chromatography as an adjunct to their identification when grown on blood agar at 35 degrees C. The organisms could be divided into two groups. In the first group (branched-chain type), which included coryneform CDC groups A-3, A-4, and A-5; some strains of B-1 and B-3; "Corynebacterium aquaticum"; Brevibacterium liquefaciens; Rothia dentocariosa; and Listeria spp., the rods had sizable quantities of antiesopentadecanoic (Ca15:0) and anteisoheptadecanoic (Ca17:0) acids. Other species with these types of CFA included B. acetylicum, which contained large amounts of isotridecanoic (Ci13:0) and anteisotridecanoic (Ca13:0) acids. CFAs useful for distinguishing among Jonesia denitrificans, Oerskovia spp., some strains of CDC groups B-1 and B-3, Kurthia spp., and Propionibacterium avidum were hexadecanoic (C 16:0) acid, isopentadecanoic (Ci15:0) acid, and Ca15:0). The second group (straight-chained type), which included Actinomyces pyogenes; Arcanobacterium haemolyticum; C. bovis; C. cystitidis; C. diphtheriae; C. flavescens, "C. gentalium"; C. jeikeium; C. kutscheri; C. matruchotii; C .minutissimum; C. mycetoides; C. pilosum; C. pseudodiphtheriticum; "C. pseudogenitalium"; C. pseudotuberculosis; C. renale; CDC groups 1, 2, ANF-1, D-2, E, F-1, F-2, G-1, G-2, and I-2; C. striatum; "C. tuberculostearicum"; C. ulcerans; C. vitarumen; C. xerosis; and Erysipelothrix rhusiopathiae, was typified by significant quantities of hexadecanoic (C16:0) and oleic acids (C18:cis9), with differences in the amounts of linoleic acid (C18:2), stearic acid (C18:0), an unnamed peak (equivalent chain length, 14.966), and small quantities of other known saturated and unsaturated fatty acids. CFA composition of these organisms was sufficiently discriminatory to assist in classification but could not be used as the sole means of identification.

Bernard, K A; Bellefeuille, M; Ewan, E P

1991-01-01

233

Identification of a new family of enzymes with potential O-acetylpeptidoglycan esterase activity in both Gram-positive and Gram-negative bacteria  

PubMed Central

Background The metabolism of the rigid bacterial cell wall heteropolymer peptidoglycan is a dynamic process requiring continuous biosynthesis and maintenance involving the coordination of both lytic and synthetic enzymes. The O-acetylation of peptidoglycan has been proposed to provide one level of control on these activities as this modification inhibits the action of the major endogenous lytic enzymes, the lytic transglycosylases. The O-acetylation of peptidoglycan also inhibits the activity of the lysozymes which serve as the first line of defense of host cells against the invasion of bacterial pathogens. Despite this central importance, there is a dearth of information regarding peptidoglycan O-acetylation and nothing has previously been reported on its de-acetylation. Results Homology searches of the genome databases have permitted this first report on the identification of a potential family of O-Acetylpeptidoglycan esterases (Ape). These proteins encoded in the genomes of a variety of both Gram-negative and Gram-positive bacteria, including a number of important human pathogens such as species of Neisseria, Helicobacter, Campylobacter, and Bacillus anthracis, have been organized into three families based on amino acid sequence similarities with family 1 being further divided into three sub-families. The genes encoding these proteins are shown to be clustered with Peptidoglycan O-acetyltransferases (Pat) and in some cases, together with other genes involved in cell wall metabolism. Representative bacteria that encode the Ape proteins were experimentally shown to produce O-acetylated peptidoglycan. Conclusion The hypothetical proteins encoded by the pat and ape genes have been organized into families based on sequence similarities. The Pat proteins have sequence similarity to Pseudomonas aeruginosa AlgI, an integral membrane protein known to participate in the O-acetylation of the exopolysaccaride, alginate. As none of the bacteria that harbor the pat genes produce alginate, we propose that the Pat proteins serve to O-acetylate peptidoglycan which is known to be a maturation event occurring in the periplasm. The Ape sequences have amino acid sequence similarity to the CAZy CE 3 carbohydrate esterases, a family previously known to be composed of only O-acetylxylan esterases. They are predicted to contain the ?/? hydrolase fold associated with the GDSL and TesA hydrolases and they possess the signature motifs associated with the catalytic residues of the CE3 esterases. Specific signature sequence motifs were identified for the Ape proteins which led to their organization into distinct families. We propose that by expressing both Pat and Ape enzymes, bacteria would be able to obtain a high level of localized control over the degradation of peptidoglycan through the attachment and removal of O-linked acetate. This would facilitate the efficient insertion of pores and flagella, localize spore formation, and control the level of general peptidoglycan turnover.

Weadge, Joel T; Pfeffer, John M; Clarke, Anthony J

2005-01-01

234

Thinking about Bacillus subtilis as a multicellular organism.  

PubMed

Initial attempts to use colony morphogenesis as a tool to investigate bacterial multicellularity were limited by the fact that laboratory strains often have lost many of their developmental properties. Recent advances in elucidating the molecular mechanisms underlying colony morphogenesis have been made possible through the use of undomesticated strains. In particular, Bacillus subtilis has proven to be a remarkable model system to study colony morphogenesis because of its well-characterized developmental features. Genetic screens that analyze mutants defective in colony morphology have led to the discovery of an intricate regulatory network that controls the production of an extracellular matrix. This matrix is essential for the development of complex colony architecture characterized by aerial projections that serve as preferential sites for sporulation. While much progress has been made, the challenge for future studies will be to determine the underlying mechanisms that regulate development such that differentiation occurs in a spatially and temporally organized manner. PMID:17977783

Aguilar, Claudio; Vlamakis, Hera; Losick, Richard; Kolter, Roberto

2007-12-01

235

Horizontal spread of mer operons among Gram-positive bacteria in natural environments  

Microsoft Academic Search

Horizontal dissemination of the genes responsible for resistance to toxic pollutants may play a key role in the adaptation of bacterial populations to environmental contaminants. However, the frequency and extent of gene dissemination in natural environments is not known. A natural horizontal spread of two distinct mercury resistance (mer) operon variants, which occurred amongst diverse Bacillus and related species over

E. S. Bogdanova; I. A. Bass; L. S. Minakhin; M. A. Petrova; S. Z. Mindlin; A. A. Volodin; E. S. Kalyaeva; J. M. Tiedje; J. L. Hobman; N. L. Brown; V. G. Nikiforov

1998-01-01

236

Antibiotic susceptibilities of Gram-positive anaerobic cocci: results of a sentinel study in England and Wales  

Microsoft Academic Search

Objective: A sentinel study was carried out to determine the antimicrobial susceptibilities of Gram-positive anaerobic cocci (GPAC) freshly isolated from clinical material in diagnostic laboratories in England and Wales. Methods: A total of 113 GPAC isolates consisting predominantly of current or former members of the genus Peptostreptococcus was obtained from 17 sentinel laboratories in England and one in Wales. Minimum

J. S. Brazier; T. E. Morris; M. Gal; B. I. Duerden

2003-01-01

237

Dustborne and airborne gram-positive and gram-negative bacteria in high versus low ERMI homes  

EPA Science Inventory

The study aimed at investigating Gram-positive and Gram-negative bacteria in moldy and non-moldy homes, as defined by the home's Environmental Relative Moldiness Index (ERMI) value. The ERMI values were determined from floor dust samples in 2010 and 2011 and homes were classified...

238

Primers for overlooked nirK, qnorB, and nosZ genes of thermophilic Gram-positive denitrifiers.  

PubMed

Although efforts have been made the past few years, knowledge on genomic and phenotypic diversity and occurrence of the denitrification ability in Gram-positive bacteria are still fragmentary. Many environmental monitoring approaches have used nir, nor, and nos genes as marker genes for detection of denitrification or denitrifying bacteria. However, primers used in these methods often fail to detect the genes in specific bacterial taxa, such as Gram-positive denitrifiers. In this study, novel primer sets specifically targeting nirK, qnorB, and nosZ genes of the Firmicute genus Geobacillus were developed by genomic mining and tested in parallel with commonly used primers on a set of phylogenetically closely related denitrifying geobacilli. Novel nirK and qnorB sequences were recovered from all strains tested, whereas nosZ was detected in part of the strain set, which was in agreement with observed phenotypes. Interspecies and modest intraspecies variations in amplified fragment length polymorphism (AFLP) patterns were observed, verifying presence of genomic variation within the strain set. Our study shows that closely related Gram-positive denitrifiers may differ in denitrification phenotype and genotype. But foremost, novel primers targeting very divergent nirK, qnorB, and nosZ gene sequences of Gram-positive denitrifiers, are now available for cultivation-independent environmental surveys. PMID:24784780

Verbaendert, Ines; Hoefman, Sven; Boeckx, Pascal; Boon, Nico; De Vos, Paul

2014-07-01

239

In Vitro Activities of a New Lipopeptide, HMR 1043, against Susceptible and Resistant Gram-Positive Isolates  

Microsoft Academic Search

The purpose of this study was to compare the activity of HMR 1043 with those of daptomycin and teicoplanin against gram-positive isolates. Susceptibility tests were performed for 52 strains, 26 parental strains, including staphylococcal, streptococcal, enterococcal, and listerial strains, and 26 HMR 1043-resistant mutants obtained from parental strains by using the Szybalski method. Agar dilution and disk diffusion susceptibility tests

Pascale Bemer; Marie-Emmanuelle Juvin; Andre Bryskier; Henri Drugeon

2003-01-01

240

Comparative Surveillance Study of Telavancin Activity against Recently Collected Gram-Positive Clinical Isolates from across the United States  

Microsoft Academic Search

Telavancin is an investigational, rapidly bactericidal lipoglycopeptide antibiotic that is being developed to treat serious infections caused by gram-positive bacteria. A baseline prospective surveillance study was conducted to assess telavancin activity, in comparison with other agents, against contemporary clinical isolates collected from 2004 to 2005 from across the United States. Nearly 4,000 isolates were collected, including staphylococci, enterococci, and streptococci

Deborah C. Draghi; Bret M. Benton; Kevin M. Krause; Clyde Thornsberry; Chris Pillar; Daniel F. Sahm

2008-01-01

241

Lipoteichoic acid preparations of gram-positive bacteria induce interleukin-12 through a CD14-dependent pathway.  

PubMed Central

Interleukin 12 (IL-12) strongly augments gamma interferon production by natural killer (NK) and T cells. IL-12 also promotes effective cell-mediated immune responses, which are particularly important against intracellular bacteria such as Listeria monocytogenes. While the lipopolysaccharide (LPS) of gram-negative bacteria induces monocyte production of IL-12, the relevant gram-positive components which induce IL-12 production are uncharacterized. We used the human monocytic cell line THP-1 to study IL-12 induction by gram-positive bacteria. Muramyl dipeptides as well as the major muramyl tetrapeptide component of Streptococcus pneumoniae were inactive for inducing IL-12. In contrast, lipoteichoic acid (LTA), a predominant surface glycolipid of gram-positive bacteria, potently induced IL-12 p40 gene expression. A competitive LPS antagonist, Rhodobacter sphaeroides LPS, inhibited LTA-induced IL-12 production, suggesting a common pathway for LPS and LTA in IL-12 activation. Pretreatment of cells with anti-CD14 monoclonal antibody blocked both LPS and LTA induction of IL-12 p40 expression. LTA also induced Thl development in naive CD4 T cells by an IL-12-dependent mechanism, indicating direct induction of physiologic levels of IL-12. Together, these results show that LTA is a potent surface structure of gram-positive bacteria which induces IL-12 in monocytes through a CD14-mediated pathway.

Cleveland, M G; Gorham, J D; Murphy, T L; Tuomanen, E; Murphy, K M

1996-01-01

242

Quantification of Gram-positive bacteria: adaptation and evaluation of a preparation strategy using high amounts of clinical tissue  

PubMed Central

Background A preparation method for quantification of bacteria in tissues is obligatory to reduce tissue mass, concentrate the target, purify, remove inhibitory substances and to achieve constant target recovery rates. No preparation method has been available until now for a high mass of tissue applicable for routine use and analytical veterinary diagnostics. Results This study describes an easy-to-use tissue preparation protocol to quantify Gram-positive bacteria from a large volume of tissue matrix. A previously published sample preparation method (Matrix-Lysis) from food science was successfully adapted for clinical use on tissues from pigs, including cerebrum, spinal cord, lung, liver, ileum, colon, caecum, kidney and muscle tissue. This tissue preparation method now permits quantification of pathogens from 5 g of organic matrix, which is a 20–200 fold increase by weight compared to other methods. It is based on solubilization of the sample matrix with either a chaotrope plus detergent or divalent salts as solubilization agents. The method was designed as a modular system, offering the possibility to change lysis buffers, according to tissue solubilization characteristics and the intended detection method (molecular or culture). Using Listeria monocytogenes as model organism, viable cell quantification or DNA extraction and quantitative real-time PCR were performed after Matrix-Lysis to determine recovery rates and detection limit (LOD). The adapted Matrix-Lysis protocol resulted in high recovery rates (mean value: 76%?±?39%) for all tested organs, except kidney, and recovery was constant over 5 log scales for all tested buffer systems. The LOD for Matrix-Lysis with subsequent plate count method (PCM) was as low as 1 CFU/5 g, while for qPCR based detection the LOD was 102 bacterial cell equivalents (BCE)/5 g for two buffer systems. Conclusions This tissue preparation is inexpensive and can be easily used for routine and analytical veterinary diagnostics. Inoculation studies or hazard assessments can profit from this tissue preparation method and it is anticipated that this study will be a valuable source for further research on tissue preparation strategies.

2014-01-01

243

Plantazolicin, a Novel Microcin B17/Streptolysin S-Like Natural Product from Bacillus amyloliquefaciens FZB42 ?  

PubMed Central

Here we report on a novel thiazole/oxazole-modified microcin (TOMM) from Bacillus amyloliquefaciens FZB42, a Gram-positive soil bacterium. This organism is well known for stimulating plant growth and biosynthesizing complex small molecules that suppress the growth of bacterial and fungal plant pathogens. Like microcin B17 and streptolysin S, the TOMM from B. amyloliquefaciens FZB42 undergoes extensive posttranslational modification to become a bioactive natural product. Our data show that the modified peptide bears a molecular mass of 1,335 Da and displays antibacterial activity toward closely related Gram-positive bacteria. A cluster of 12 genes that covers ?10 kb is essential for the production, modification, export, and self-immunity of this natural product. We have named this compound plantazolicin (PZN), based on the association of several producing organisms with plants and the incorporation of azole heterocycles, which derive from Cys, Ser, and Thr residues of the precursor peptide.

Scholz, Romy; Molohon, Katie J.; Nachtigall, Jonny; Vater, Joachim; Markley, Andrew L.; Sussmuth, Roderich D.; Mitchell, Douglas A.; Borriss, Rainer

2011-01-01

244

Evaluation of the Andromas matrix-assisted laser desorption ionization-time of flight mass spectrometry system for identification of aerobically growing Gram-positive bacilli.  

PubMed

Matrix-associated laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a rapid and simple microbial identification method. Previous reports using the Biotyper system suggested that this technique requires a preliminary extraction step to identify Gram-positive rods (GPRs), a technical issue that may limit the routine use of this technique to identify pathogenic GPRs in the clinical setting. We tested the accuracy of the MALDI-TOF MS Andromas strategy to identify a set of 659 GPR isolates representing 16 bacterial genera and 72 species by the direct colony method. This bacterial collection included 40 C. diphtheriae, 13 C. pseudotuberculosis, 19 C. ulcerans, and 270 other Corynebacterium isolates, 32 L. monocytogenes and 24 other Listeria isolates, 46 Nocardia, 75 Actinomyces, 18 Actinobaculum, 11 Propionibacterium acnes, 18 Propionibacterium avidum, 30 Lactobacillus, 21 Bacillus, 2 Rhodococcus equi, 2 Erysipelothrix rhusiopathiae, and 38 other GPR isolates, all identified by reference techniques. Totals of 98.5% and 1.2% of non-Listeria GPR isolates were identified to the species or genus level, respectively. Except for L. grayi isolates that were identified to the species level, all other Listeria isolates were identified to the genus level because of highly similar spectra. These data demonstrate that rapid identification of pathogenic GPRs can be obtained without an extraction step by MALDI-TOF mass spectrometry. PMID:22692743

Farfour, E; Leto, J; Barritault, M; Barberis, C; Meyer, J; Dauphin, B; Le Guern, A-S; Leflèche, A; Badell, E; Guiso, N; Leclercq, A; Le Monnier, A; Lecuit, M; Rodriguez-Nava, V; Bergeron, E; Raymond, J; Vimont, S; Bille, E; Carbonnelle, E; Guet-Revillet, H; Lécuyer, H; Beretti, J-L; Vay, C; Berche, P; Ferroni, A; Nassif, X; Join-Lambert, O

2012-08-01

245

The Terminally Redundant, Nonpermuted Genome of Listeria Bacteriophage A511: a Model for the SPO1-Like Myoviruses of Gram-Positive Bacteria? †  

PubMed Central

Only little information on a particular class of myoviruses, the SPO1-like bacteriophages infecting low-G+C-content, gram-positive host bacteria (Firmicutes), is available. We present the genome analysis and molecular characterization of the large, virulent, broad-host-range Listeria phage A511. A511 contains a unit (informational) genome of 134,494 bp, encompassing 190 putative open reading frames (ORFs) and 16 tRNA genes, organized in a modular fashion common among the Caudovirales. Electron microscopy, enzymatic fragmentation analyses, and sequencing revealed that the A511 DNA molecule contains linear terminal repeats of a total of 3,125 bp, encompassing nine small putative ORFs. This particular genome structure explains why A511 is unable to perform general transduction. A511 features significant sequence homologies to Listeria phage P100 and other morphologically related phages infecting Firmicutes such as Staphylococcus phage K and Lactobacillus phage LP65. Equivalent but more-extensive terminal repeats also exist in phages P100 (?6 kb) and K (?20 kb). High-resolution electron microscopy revealed, for the first time, the presence of long tail fibers organized in a sixfold symmetry in these viruses. Mass spectrometry-based peptide fingerprinting permitted assignment of individual proteins to A511 structural components. On the basis of the data available for A511 and relatives, we propose that SPO1-like myoviruses are characterized by (i) their infection of gram-positive, low-G+C-content bacteria; (ii) a wide host range within the host bacterial genus and a strictly virulent lifestyle; (iii) similar morphology, sequence relatedness, and collinearity of the phage genome organization; and (iv) large double-stranded DNA genomes featuring nonpermuted terminal repeats of various sizes.

Klumpp, Jochen; Dorscht, Julia; Lurz, Rudi; Bielmann, Regula; Wieland, Matthias; Zimmer, Markus; Calendar, Richard; Loessner, Martin J.

2008-01-01

246

The terminally redundant, nonpermuted genome of Listeria bacteriophage A511: a model for the SPO1-like myoviruses of gram-positive bacteria.  

PubMed

Only little information on a particular class of myoviruses, the SPO1-like bacteriophages infecting low-G+C-content, gram-positive host bacteria (Firmicutes), is available. We present the genome analysis and molecular characterization of the large, virulent, broad-host-range Listeria phage A511. A511 contains a unit (informational) genome of 134,494 bp, encompassing 190 putative open reading frames (ORFs) and 16 tRNA genes, organized in a modular fashion common among the Caudovirales. Electron microscopy, enzymatic fragmentation analyses, and sequencing revealed that the A511 DNA molecule contains linear terminal repeats of a total of 3,125 bp, encompassing nine small putative ORFs. This particular genome structure explains why A511 is unable to perform general transduction. A511 features significant sequence homologies to Listeria phage P100 and other morphologically related phages infecting Firmicutes such as Staphylococcus phage K and Lactobacillus phage LP65. Equivalent but more-extensive terminal repeats also exist in phages P100 (approximately 6 kb) and K (approximately 20 kb). High-resolution electron microscopy revealed, for the first time, the presence of long tail fibers organized in a sixfold symmetry in these viruses. Mass spectrometry-based peptide fingerprinting permitted assignment of individual proteins to A511 structural components. On the basis of the data available for A511 and relatives, we propose that SPO1-like myoviruses are characterized by (i) their infection of gram-positive, low-G+C-content bacteria; (ii) a wide host range within the host bacterial genus and a strictly virulent lifestyle; (iii) similar morphology, sequence relatedness, and collinearity of the phage genome organization; and (iv) large double-stranded DNA genomes featuring nonpermuted terminal repeats of various sizes. PMID:18567664

Klumpp, Jochen; Dorscht, Julia; Lurz, Rudi; Bielmann, Regula; Wieland, Matthias; Zimmer, Markus; Calendar, Richard; Loessner, Martin J

2008-09-01

247

Intersection of the stringent response and the CodY regulon in low GC Gram-positive bacteria.  

PubMed

Bacteria adapt efficiently to a wide range of nutritional environments. Therefore, they possess overlapping regulatory systems that detect intracellular pools of key metabolites. In low GC Gram-positive bacteria, two global regulators, the stringent response and the CodY repressor, respond to an intracellular decrease in amino acid content. Amino acid limitation leads to rapid synthesis of the alarmones pppGpp and ppGpp through the stringent response and inactivates the CodY repressor. Two cofactors, branched chain amino acids (BCAA) and GTP, are ligands for CodY and facilitate binding to the target DNA. Because (p)ppGpp synthesis and accumulation evidentially reduce the intracellular GTP pool, CodY is released from the DNA, and transcription of target genes is altered. Here, we focus on this intimate link between the stringent response and CodY regulation in different Gram-positive species. PMID:24462007

Geiger, Tobias; Wolz, Christiane

2014-03-01

248

Identification of lactic acid bacteria and Gram-positive catalase-positive cocci isolated from naturally fermented sausage (sucuk).  

PubMed

The aim of the study was to identify lactic acid bacteria and Gram-positive catalase-positive cocci isolated from Turkish dry fermented sausage (sucuk) produced by 7 different manufacturers without using starter culture. A total of 129 isolates of lactic acid bacteria were identified phenotypically. Lactobacillus plantarum was the dominant species (45.7%) followed by L. curvatus (10.9%) and L. fermentum (9.3%). Pediococcus isolates were identified as P. pentosaceus and P. acidilactici. All the isolates of gram-positive and catalase-positive cocci (123 isolates) were classified as Staphylococcus except for 1 isolate assigned to Kocuria rosea. The species isolated most often were S. xylosus (41.5%) and S. saprophyticus (28.5%). Four isolates were identified as S. equorum (3.3%), 1 isolate was assigned to S. carnosus (0.8%). PMID:19019118

Kaban, G; Kaya, M

2008-10-01

249

In vitro activities of a new lipopeptide, HMR 1043, against susceptible and resistant gram-positive isolates.  

PubMed

The purpose of this study was to compare the activity of HMR 1043 with those of daptomycin and teicoplanin against gram-positive isolates. Susceptibility tests were performed for 52 strains, 26 parental strains, including staphylococcal, streptococcal, enterococcal, and listerial strains, and 26 HMR 1043-resistant mutants obtained from parental strains by using the Szybalski method. Agar dilution and disk diffusion susceptibility tests were performed by the procedures outlined by the NCCLS. HMR 1043 demonstrated good activity against susceptible and resistant gram-positive bacteria. The activity of HMR 1043 in vitro was less influenced by the presence of calcium ions than that of daptomycin. Susceptibility test breakpoints were not defined because of the poor correlation coefficients obtained with the different disks tested. PMID:12937020

Bemer, Pascale; Juvin, Marie-Emmanuelle; Bryskier, Andre; Drugeon, Henri

2003-09-01

250

In Vitro Activities of a New Lipopeptide, HMR 1043, against Susceptible and Resistant Gram-Positive Isolates  

PubMed Central

The purpose of this study was to compare the activity of HMR 1043 with those of daptomycin and teicoplanin against gram-positive isolates. Susceptibility tests were performed for 52 strains, 26 parental strains, including staphylococcal, streptococcal, enterococcal, and listerial strains, and 26 HMR 1043-resistant mutants obtained from parental strains by using the Szybalski method. Agar dilution and disk diffusion susceptibility tests were performed by the procedures outlined by the NCCLS. HMR 1043 demonstrated good activity against susceptible and resistant gram-positive bacteria. The activity of HMR 1043 in vitro was less influenced by the presence of calcium ions than that of daptomycin. Susceptibility test breakpoints were not defined because of the poor correlation coefficients obtained with the different disks tested.

Bemer, Pascale; Juvin, Marie-Emmanuelle; Bryskier, Andre; Drugeon, Henri

2003-01-01

251

In Vitro Activity of Ozenoxacin against Quinolone-Susceptible and Quinolone-Resistant Gram-Positive Bacteria  

PubMed Central

In vitro activity of ozenoxacin, a novel nonfluorinated topical (L. D. Saravolatz and J. Leggett, Clin. Infect. Dis. 37:1210–1215, 2003) quinolone, was compared with the activities of other quinolones against well-characterized quinolone-susceptible and quinolone-resistant Gram-positive bacteria. Ozenoxacin was 3-fold to 321-fold more active than other quinolones. Ozenoxacin could represent a first-in-class nonfluorinated quinolone for the topical treatment of a broad range of dermatological infections.

Lopez, Y.; Tato, M.; Espinal, P.; Garcia-Alonso, F.; Gargallo-Viola, D.; Canton, R.

2013-01-01

252

In Vitro Activities of Linezolid against Important Gram-Positive Bacterial Pathogens Including Vancomycin-Resistant Enterococci  

Microsoft Academic Search

The emergence of resistance in gram-positive bacteria has necessitated a search for new antimicrobial agents. Linezolid is an oxazolidinone, a new class of antibacterial agents with enhanced activity against pathogens. We compared the activity of linezolid to those of other antimicrobial agents against 3,945 clinical isolates. Linezolid demonstrated potent activity against all isolates tested. For all vancomycin-susceptible enterococci, staphylococci, and

GARY A. NOSKIN; FARIDA SIDDIQUI; VALENTINA STOSOR; DONNA HACEK; LANCE R. PETERSON

1999-01-01

253

Comparison of the D-Glutamate-Adding Enzymes from Selected Gram-Positive and Gram-Negative Bacteria  

Microsoft Academic Search

The biochemical properties of the D-glutamate-adding enzymes (MurD) from Escherichia coli, Haemophilus influenzae, Enterococcus faecalis, and Staphylococcus aureus were investigated to detect any differences in the activity of this enzyme between gram-positive and gram-negative bacteria. The genes (murD) that encode these enzymes were cloned into pMAL-c2 fusion vector and overexpressed as maltose-binding protein-MurD fusion proteins. Each fusion protein was purified

ANN W. WALSH; PAUL J. FALK; JANE THANASSI; LINDA DISCOTTO; MICHAEL J. PUCCI; HSU-TSO HO

1999-01-01

254

Role of toll-like receptor 4 in gram-positive and gram-negative pneumonia in mice  

Microsoft Academic Search

To determine the role of Toll-like receptor 4 (TLR4) in the immune response to pneumonia, C3H\\/HeJ mice (which display a mutant nonfunctional TLR4) and C3H\\/HeN wild-type mice were intranasally infected with either Streptococcus pneumoniae (a common gram-positive respiratory pathogen) or Klebsiella pneumoniae (a common gram-negative respiratory pathogen). In cases of pneumococcal pneumonia, TLR4 mutant mice showed a reduced survival only

Judith Branger; Sylvia Knapp; Sebastiaan Weijer; Jaklien C. Leemans; Pater de J. M; Peter Speelman; Sandrine Florquin; Tom van der Poll

2004-01-01

255

Sustained generation of electricity by the spore-forming, Gram-positive, Desulfitobacterium hafniense strain DCB2  

Microsoft Academic Search

Desulfitobacterium hafniense strain DCB2 gen- erates electricity in microbial fuel cells (MFCs) when humic acids or the humate analog anthraquinone-2,6- disulfonate (AQDS) is added as an electron-carrying mediator. When utilizing formate as fuel, the Gram- positive, spore-forming bacterium generated up to 400 mW\\/m2 of cathode surface area in a single-chamber MFC with a platinum-containing air-fed cathode. Hydro- gen, lactate, pyruvate,

C. E. Milliken; H. D. May

2007-01-01

256

Evaluation of the VITEK 2 System for Identification and Antimicrobial Susceptibility Testing of Medically Relevant Gram-Positive Cocci  

Microsoft Academic Search

A study was conducted to evaluate the new VITEK 2 system (bioMérieux) for identification and antibiotic susceptibility testing of gram-positive cocci. Clinical isolates of Staphylococcus aureus (n 100), coagulase- negative staphylococci (CNS) (n 100), Enterococcus spp. (n 89), Streptococcus agalactiae (n 29), and Streptococcus pneumoniae (n 66) were examined with the ID-GPC identification card and with the AST-P515 (for staphylococci),

Marco Ligozzi; Cinzia Bernini; Maria Grazia Bonora; Maria de Fatima; Jessica Zuliani; Roberta Fontana

2002-01-01

257

Spectrum and potency of dalbavancin tested against 3322 Gram-positive cocci isolated in the United States Surveillance Program (2004)  

Microsoft Academic Search

Dalbavancin is an injectable, next generation lipoglycopeptide with an extended serum elimination half-life. Once-weekly dosing has been successful for treatment of skin and skin structure (SSSI) and catheter-related bloodstream infections (CR-BSI). Concurrent with clinical trails, dalbavancin resistance surveillance was initiated in 2003, and results are reported here for the 2004 United States (USA) component. A total of 3322 Gram-positive cocci

Ronald N. Jones; Matthew G. Stilwell; Helio S. Sader; Thomas R. Fritsche; Beth P. Goldstein

2006-01-01

258

Dustborne and airborne Gram-positive and Gram-negative bacteria in high versus low ERMI homes.  

PubMed

The study aimed at investigating Gram-positive and Gram-negative bacteria in moldy and non-moldy homes, as defined by the home's Environmental Relative Moldiness Index (ERMI) value. The ERMI values were determined from floor dust samples in 2010 and 2011 and homes were classified into low (<5) and high (>5) ERMI groups based on the average ERMI values as well as 2011 ERMI values. Dust and air samples were collected from the homes in 2011 and all samples were analyzed for Gram-positive and Gram-negative bacteria using QPCR assays, endotoxin by the LAL assay, and N-acetyl-muramic acid using HPLC. In addition, air samples were analyzed for culturable bacteria. When average ERMI values were considered, the concentration and load of Gram-positive bacteria determined with QPCR in house dust, but not air, were significantly greater in high ERMI homes than in low ERMI homes. Furthermore, the concentration of endotoxin, but not muramic acid, in the dust was significantly greater in high ERMI than in low ERMI homes. In contrast, when ERMI values of 2011 were considered, Gram-negative bacteria determined with QPCR in air, endotoxin in air, and muramic acid in dust were significantly greater in high ERMI homes. The results suggest that both short-term and long-term mold contamination in homes could be linked with the bacterial concentrations in house dust, however, only the current mold status was associated with bacterial concentrations in air. Although correlations were found between endotoxin and Gram-negative bacteria as well as between muramic acid and Gram-positive bacteria in the entire data set, diverging associations were observed between the different measures of bacteria and the home moldiness. It is likely that concentrations of cells obtained by QPCR and concentrations of cell wall components are not equivalent and represent too broad categories to understand the bacterial composition and sources of the home microbiota. PMID:24642096

Adhikari, Atin; Kettleson, Eric M; Vesper, Stephen; Kumar, Sudhir; Popham, David L; Schaffer, Christopher; Indugula, Reshmi; Chatterjee, Kanistha; Allam, Karteek K; Grinshpun, Sergey A; Reponen, Tiina

2014-06-01

259

Meso-substituted cationic porphyrins as efficient photosensitizers of gram-positive and gram-negative bacteria  

Microsoft Academic Search

Previous studies on the photosensitization of bacterial cells with different neutral or negatively charged porphyrins and phthalocyanines have demonstrated that, although Gram-positive bacteria are efficiently photoinactivated, Gram-negatrive bacteria become photosensitive only after modification of the permeability of their outer membrane.The results described in this paper show that two meso-substituted cationic porphyrins, namely tetra(4N-methyl-pyridyl) porphine tetraiodide and the tetra(4N,N,N,-trimethyl-anilinium) porphine, efficiently

Michèle Merchat; Giulio Bertolini; Paolo Giacomini; Angeles Villaneuva; Giulio Jori

1996-01-01

260

Coexistence of CD14Dependent and Independent Pathways for Stimulation of Human Monocytes by Gram-Positive Bacteria  

Microsoft Academic Search

The cell wall is a key inflammatory agent of gram-positive bacteria. Possible receptors mediating cell wall-induced inflammation include CD14 and platelet-activating factor (PAF) receptor. To delineate the conditions under which these various receptors might be used, human monocytic THP-1 cells and heparinized whole human blood were stimulated with lipopolysaccharide (LPS), intact Streptococcus pneumoniae bacteria, or purified pneumococcal cell wall. THP-1

ANJE CAUWELS; ELAINE WAN; MICHAELA LEISMANN; ELAINE TUOMANEN

1997-01-01

261

Improved Pattern for Genome-Based Screening Identifies Novel Cell Wall-Attached Proteins in Gram-Positive Bacteria  

Microsoft Academic Search

With a large number of sequenced microbial genomes available, tools for identifying groups or classes of proteins have become increasingly important. Here we present an improved pattern for the identification of cell wall-attached proteins (CWPs), a group of proteins with diverse and important functions in gram-positive bacteria. This tripartite pattern is based on analysis of 65 previously described cell wall-attached

ROBERT JANULCZYK; MAGNUS RASMUSSEN

2001-01-01

262

Com parative genomics of the methionine metabolism in Gram-positive bacteria: a variety of regulatory systems  

Microsoft Academic Search

Regulation of the methionine biosynthesis and trans- port genes in bacteria is rather diverse and involves two RNA-level regulatory systems and at least three DNA-level systems. In particular, the methionine metabolism in Gram-positive bacteria was known to be controlled by the S-box and T-box mechanisms, both acting on the level of premature termination of transcription. Using comparative analysis of genes,

Dmitry A. Rodionov; Alexey G. Vitreschak; Andrey A. Mironov; Mikhail S. Gelfand

263

Effects of cell structure of gram-positive and gram-negative bacteria based on their dielectric properties  

Microsoft Academic Search

Current procedures for the identification and characterization of microorganisms are based on complex and time-consuming protocols. Here, we hypothesize that the distinct cellular composition of microorganisms could be identified by dielectric spectroscopy. Gram-positive and Gram-negative bacterial strain permittivity measurement was performed using a network analyzer and a coaxial probe in the frequency range from 50 MHz to 300 MHz. All

Katja Dahlke; Christiane Geyer; Stephan Dees; Marko Helbig; Jurgen Sachs; Francesco Scotto di Clemente; Matthias Hein; Werner Alois Kaiser; Ingrid Hilger

2012-01-01

264

Studies on the aminopeptidase test for the distinction of gram-negative from gram-positive bacteria  

Microsoft Academic Search

The aminopeptidase test was performed with representatives of gram-negative, gram-positive, and gram-variable bacteria. All gram-negative bacteria tested gave a positive test reaction with L-alanine-4-nitroanilide as test substrate. Representatives of the coryneform bacteria and some streptococci showed aminopeptidase activity after prolonged reaction times. A correlation between aminopeptidase activity and distinct interpeptide bridge composition in the peptidoglycan of many strains was demonstrated.

G. Cerny

1978-01-01

265

[Evaluation of API Coryne System, version 2.0, for diphteroid gram-positive rods identification with clinical relevance].  

PubMed

The ability of the API Coryne system, version 2.0, to identify 178 strains of gram-positive rods was evaluated. Seventy eight isolates belonged to genus Corynebacterium and one hundred to related genera, all strains were isolated from clinical samples at the Laboratory of Bacteriology, Hospital de Clínicas José de San Martin (UBA) between 1995 and 2004. The isolates were identified according to von Graevenitz and Funke's scheme. One hundred and sixty two out of 178 strains (91%) were correctly identified at genus and species level (IC95 = 85.6-94.6), in 44 of them (24.7%) additional tests were needed to final identification. Sixteen strains (9%) were not correctly identified (IC95 = 5.4-14.4); none of the 178 strains remained unidentified. The API Coryne system, version 2.0, is useful to identify the majority of Cory-nebacterium species with clinical relevance: Corynebacterium jeikeium, Corynebacterium urealyticum, Corynebacterium striatum, Corynebacterium pseudodiphtheriticum, Corynebacterium amycolatum and related species such as Arcanobacterium haemolyticum, Dermabacter hominis, Listeria monocytogenes, among others. Nevertheless for yellow-pigmented diphteroid gram-positive rods (Aureobacterium spp., Leifsonia aquatica, Microbacterium spp. and Cellulomonas spp.) and for acid fast gram-positive rods (Rhodococcus, Gordonia, Tsukamurella and Nocardia) the identification usefulness the system is limited. PMID:17370571

Almuzara, M N; De Mier, C; Rodríguez, C R; Famiglietti, A M R; Vay, C A

2006-01-01

266

Probing the relationship between Gram-negative and Gram-positive S1 proteins by sequence analysis  

PubMed Central

Escherichia coli ribosomal protein S1 is required for the translation initiation of messenger RNAs, in particular when their Shine–Dalgarno sequence is degenerated. Closely related forms of the protein, composed of the same number of domains (six), are found in all Gram-negative bacteria. More distant proteins, generally formed of fewer domains, have been identified, by sequence similarities, in Gram-positive bacteria and are also termed ‘S1 proteins’. However in the absence of functional information, it is generally difficult to ascertain their relationship with Gram-negative S1. In this article, we report the solution structure of the fourth and sixth domains of the E. coli protein S1 and show that it is possible to characterize their ?-barrel by a consensus sequence that allows a precise identification of all domains in Gram-negative and Gram-positive S1 proteins. In addition, we show that it is possible to discriminate between five domain types corresponding to the domains 1, 2, 3, 4–5 and 6 of E. coli S1 on the basis of their sequence. This enabled us to identify the nature of the domains present in Gram-positive proteins and, subsequently, to probe the filiations between all forms of S1.

Salah, Philippe; Bisaglia, Marco; Aliprandi, Pascale; Uzan, Marc; Sizun, Christina; Bontems, Francois

2009-01-01

267

Surface multiheme c-type cytochromes from Thermincola potens: Implications for dissimilatory metal reduction by Gram-positive bacteria  

NASA Astrophysics Data System (ADS)

Almost nothing is known about the mechanisms of dissimilatory metal reduction by Gram-positive bacteria, although they have been shown to be the dominant species in some environments. Thermincola potens strain JR was isolated from the anode of a microbial fuel cell inoculated with anaerobic digester sludge and operated at 55 °C. Preliminary characterization revealed that T. potens coupled acetate oxidation to the reduction of hydrous ferric oxides (HFO) or the humic substances analog, anthraquinone-2,6-disulfonate (AQDS). The genome of T. potens was recently sequenced, and the abundance of multiheme c-type cytochromes (MHCs) is unusual for a Gram-positive bacterium. We present evidence from trypsin shaving LC-MS/MS experiments and surface-enhanced Raman spectroscopy (SERS) that indicates the expression of a number of MHCs during T. potens growth on either HFO or AQDS and that several MHCs are localized to the cell wall or cell surface of T. potens. Furthermore, one of the MHCs can be extracted from cells with low pH or denaturants suggesting a loose association with the cell wall or cell surface. Electron microscopy does not reveal an S-layer, and the precipitation of silver metal on the cell surface is inhibited by cyanide, supporting the involvement of surface-localized redox-active heme proteins in dissimilatory metal reduction. These results are the first direct evidence for cell-wall associated cytochromes and MHC involvement in conducting electrons across the cell envelope of a Gram-positive bacterium.

Carlson, H. K.; Iavarone, A. T.; Gorur, A.; Yeo, B. S.; Tran, R.; Melnyk, R. A.; Mathies, R. A.; Auer, M.; Coates, J. D.

2011-12-01

268

Surface multiheme c-type cytochromes from Thermincola potens and implications for respiratory metal reduction by Gram-positive bacteria  

PubMed Central

Almost nothing is known about the mechanisms of dissimilatory metal reduction by Gram-positive bacteria, although they may be the dominant species in some environments. Thermincola potens strain JR was isolated from the anode of a microbial fuel cell inoculated with anaerobic digester sludge and operated at 55 °C. Preliminary characterization revealed that T. potens coupled acetate oxidation to the reduction of hydrous ferric oxides (HFO) or anthraquinone-2,6-disulfonate (AQDS), an analog of the redox active components of humic substances. The genome of T. potens was recently sequenced, and the abundance of multiheme c-type cytochromes (MHCs) is unusual for a Gram-positive bacterium. We present evidence from trypsin-shaving LC-MS/MS experiments and surface-enhanced Raman spectroscopy (SERS) that indicates the expression of a number of MHCs during T. potens growth on either HFO or AQDS, and that several MHCs are localized to the cell wall or cell surface. Furthermore, one of the MHCs can be extracted from cells with low pH or denaturants, suggesting a loose association with the cell wall or cell surface. Electron microscopy does not reveal an S-layer, and the precipitation of silver metal on the cell surface is inhibited by cyanide, supporting the involvement of surface-localized redox-active heme proteins in dissimilatory metal reduction. These results provide unique direct evidence for cell wall-associated cytochromes and support MHC involvement in conducting electrons across the cell envelope of a Gram-positive bacterium.

Carlson, Hans K.; Iavarone, Anthony T.; Gorur, Amita; Yeo, Boon Siang; Tran, Rosalie; Melnyk, Ryan A.; Mathies, Richard A.; Auer, Manfred; Coates, John D.

2012-01-01

269

Determination of the gram-positive bacterial content of soils and sediments by analysis of teichoic acid components  

NASA Technical Reports Server (NTRS)

Many gram-positive bacteria form substituted polymers of glycerol and ribitol phosphate esters known as teichoic acids. Utilizing the relative specificity of cold concentrated hydrofluoric acid in the hydrolysis of polyphosphate esters it proved possible to quantitatively assay the teichoic acid-derived glycerol and ribitol from gram-positive bacteria added to various soils and sediments. The lipids are first removed from the soils or sediments with a one phase chloroform-methanol extraction and the lipid extracted residue is hydrolyzed with cold concentrated hydrofluoric acid. To achieve maximum recovery of the teichoic acid ribitol, a second acid hydrolysis of the aqueous extract is required. The glycerol and ribitol are then acetylated after neutralization and analyzed by capillary gas-liquid chromatography. This technique together with measures of the total phospholipid, the phospholipid fatty acid, the muramic acid and the hydroxy fatty acids of the lipopolysaccharide lipid A of the gram-negative bacteria makes it possible to describe the community structure environmental samples. The proportion of gram-positive bacteria measured as the teichoic acid glycerol and ribitol is higher in soils than in sediments and increases with depth in both.

Gehron, M. J.; Davis, J. D.; Smith, G. A.; White, D. C.

1984-01-01

270

Development of a Treatment Algorithm for Streptococci and Enterococci from Positive Blood Cultures Identified with the Verigene Gram-Positive Blood Culture Assay  

PubMed Central

Seventy-eight blood cultures with a Gram stain result of Gram-positive cocci in pairs and/or chains were evaluated with the Nanosphere Verigene Gram-positive blood culture (BC-GP) assay. The overall concordance of the assay with culture was 89.7% (70/78 cultures), allowing for the development of a targeted treatment algorithm.

Alby, Kevin; Daniels, Lindsay M.; Weber, David J.

2013-01-01

271

Competitive adsorption of metal cations onto two gram positive bacteria: testing the chemical equilibrium model  

NASA Astrophysics Data System (ADS)

In order to test the ability of a surface complexation approach to account for metal-bacteria interactions in near surface fluid-rock systems, we have conducted experiments that measure the extent of adsorption in mixed metal, mixed bacteria systems. This study tests the surface complexation approach by comparing estimated extents of adsorption based on surface complexation modeling to those we observed in the experimental systems. The batch adsorption experiments involved Ca, Cd, Cu, and Pb adsorption onto the surfaces of 2 g positive bacteria: Bacillus subtilis and Bacillus licheniformis. Three types of experiments were performed: 1. Single metal (Ca, Cu, Pb) adsorption onto a mixture of B. licheniformis and B. subtilis; 2. mixed metal (Cd, Cu, and Pb; Ca and Cd) adsorption onto either B. subtilis or B. licheniformis; and 3. mixed or single metal adsorption onto B. subtilis and B. licheniformis. %Independent of the experimental results, and based on the site specific stability constants for Ca, Cd, Cu, and Pb interactions with the carboxyl and phosphate sites on B. licheniformis and B. subtilis determined by Fein et al. (1997), by Daughney et al. (1998) and in this study, we estimate the extent of adsorption that is expected in the above experimental systems. Competitive cation adsorption experiments in both single and double bacteria systems exhibit little adsorption at pH values less than 4. With increasing pH above 4.0, the extent of Ca, Cu, Pb and Cd adsorption also increases due to the increased deprotonation of bacterial surface functional groups. In all cases studied, the estimated adsorption behavior is in excellent agreement with the observations, with only slight differences that were within the uncertainties of the estimation and experimental procedures. Therefore, the results indicate that the use of chemical equilibrium modeling of aqueous metal adsorption onto bacterial surfaces yields accurate predictions of the distribution of metals in complex multicomponent systems.

Fowle, David A.; Fein, Jeremy B.

1999-10-01

272

Draft Genome Sequence of Bacillus thuringiensis NBIN-866 with High Nematocidal Activity  

PubMed Central

Bacillus thuringiensis NBIN-866, a Gram-positive bacterium, was isolated from soil in China. We announce here the draft genome sequence of strain B. thuringiensis NBIN-866, which possesses highly nematocidal factors, such as proteins and small molecular peptides.

Zhou, Ronghua; Fu, Guiping; Zhang, Wei; Min, Yong; Tian, Yuxi; Huang, Daye; Wang, Kaimei; Wan, Zhongyi; Yao, Jingwu; Yang, Ziwen

2014-01-01

273

Bacillus marcorestinctum sp. nov., a novel soil acylhomoserine lactone quorum-sensing signal quenching bacterium.  

PubMed

A Gram-positive, facultatively anaerobic, endospore-forming and rod-shaped bacterium was isolated from soil samples and designated strain LQQ. This organism strongly quenches the acylhomoserine lactone quorum-sensing signal. The LQQ strain exhibits phenotypic characteristics consistent with its classification in the genus Bacillus. It is positive in catalase and no special growth factor is needed. It uses glucose as sole carbon source. The DNA G + C content is 39.8 mol %. The closest relatives based on the 16S rRNA gene sequence are Bacillus anthracis, Bacillus thuringiensis, and Brevibacillus brevis (syn. Bacillus brevis) with the similarity of 96.5%. The DNA-DNA hybridization data indicates a low level of genomic relatedness with the relative type strains of Bacillus thuringiensis (6.1%), Bacillus anthracis (10.5%) and Brevibacillus brevis (8.7%). On the basis of the phenotypic and phylogenetic data together with the genomic distinctiveness, the LQQ strain represents a novel species of the genus Bacillus, for which the name Bacillus marcorestinctum sp. nov. is proposed. The type strain is LQQ(T). PMID:20386651

Han, Yan; Chen, Fang; Li, Nuo; Zhu, Bo; Li, Xianzhen

2010-01-01

274

Secretory phospholipase A2 in dromedary tears: a host defense against staphylococci and other gram-positive bacteria.  

PubMed

The best known physiologic function of secreted phospholipase A2 (sPLA2) group IIA (sPLA2-IIA) is defense against bacterial infection through hydrolytic degradation of bacterial membrane phospholipids. In fact, sPLA2-IIA effectively kills Gram-positive bacteria and to a lesser extent Gram-negative bacteria and is considered a major component of the eye's innate immune defense system. The antibacterial properties of sPLA2 have been demonstrated in rabbit and human tears. In this report, we have analyzed the bactericidal activity of dromedary tears and the subsequently purified sPLA2 on several Gram-positive bacteria. Our results showed that the sPLA2 displays a potent bactericidal activity against all the tested bacteria particularly against the Staphylococcus strains when tested in the ionic environment of tears. There is a synergic action of the sPLA2 with lysozyme when added to the bacteria culture prior to sPLA2. Interestingly, lysozyme purified from dromedary tears showed a significant bactericidal activity against Listeria monocytogene and Staphylococcus epidermidis, whereas the one purified from human tears displayed no activity against these two strains. We have also demonstrated that Ca(2+) is crucial for the activity of dromedary tear sPLA2 and to a less extent Mg(2+) ions. Given the presence of sPLA2 in tears and intestinal secretions, this enzyme may play a substantial role in innate mucosal and systemic bactericidal defenses against Gram-positive bacteria. PMID:23344945

Ben Bacha, Abir; Abid, Islem

2013-03-01

275

Comparison of the d-Glutamate-Adding Enzymes from Selected Gram-Positive and Gram-Negative Bacteria  

PubMed Central

The biochemical properties of the d-glutamate-adding enzymes (MurD) from Escherichia coli, Haemophilus influenzae, Enterococcus faecalis, and Staphylococcus aureus were investigated to detect any differences in the activity of this enzyme between gram-positive and gram-negative bacteria. The genes (murD) that encode these enzymes were cloned into pMAL-c2 fusion vector and overexpressed as maltose-binding protein–MurD fusion proteins. Each fusion protein was purified to homogeneity by affinity to amylose resin. Proteolytic treatments of the fusion proteins with factor Xa regenerated the individual MurD proteins. It was found that these fusion proteins retain d-glutamate-adding activity and have Km and Vmax values similar to those of the regenerated MurDs, except for the H. influenzae enzyme. Substrate inhibition by UDP-N-acetylmuramyl-l-alanine, the acceptor substrate, was observed at concentrations greater than 15 and 30 ?M for E. coli and H. influenzae MurD, respectively. Such substrate inhibition was not observed with the E. faecalis and S. aureus enzymes, up to a substrate concentration of 1 to 2 mM. In addition, the two MurDs of gram-negative origin were shown to require monocations such as NH4+ and/or K+, but not Na+, for optimal activity, while anions such as Cl? and SO42? had no effect on the enzyme activities. The activities of the two MurDs of gram-positive origin, on the other hand, were not affected by any of the ions tested. All four enzymes required Mg2+ for the ligase activity and exhibited optimal activities around pH 8. These differences observed between the gram-positive and gram-negative MurDs indicated that the two gram-negative bacteria may apply a more stringent regulation of cell wall biosynthesis at the early stage of peptidoglycan biosynthesis pathway than do the two gram-positive bacteria. Therefore, the MurD-catalyzed reaction may constitute a fine-tuning step necessary for the gram-negative bacteria to optimally maintain its relatively thin yet essential cell wall structure during all stages of growth.

Walsh, Ann W.; Falk, Paul J.; Thanassi, Jane; Discotto, Linda; Pucci, Michael J.; Ho, Hsu-Tso

1999-01-01

276

Molecular modeling of Gram-positive bacteria peptidoglycan layer, selected glycopeptide antibiotics and vancomycin derivatives modified with sugar moieties.  

PubMed

Proper understanding of the mechanisms of binding to Gram-positive bacteria cell wall layers-especially to the peptidoglycan (PG) layer, seems to be crucial for proper development of new drug candidates which are effective against these bacteria. In this work we have constructed two different models of the Gram-positive bacteria PG layer: the layered and the scaffold models. PG conformational changes during geometry optimization, models relaxation, and molecular dynamics were described and discussed. We have found that the border surface of both PG layer models differs from the surface located away from the edge of models and the chains formed by disaccharide units prefer helix-like conformation. This curling of PG chains significantly affects the shape of antibiotic-accessible surface and the process is thus crucial for new drug development. Glycopeptide antibiotics effective against Gram-positive bacteria, such as vancomycin and its semisynthetic derivatives-oritavancin and telavancin, bind to d-alanyl-d-alanine stem termini on the peptidoglycan precursors of the cell wall. This binding inhibits cross-linking between the peptides and subsequently prevents cell wall synthesis. In this study some of the aspects of conformational freedom of vancomycin and restrictions from the modifications of vancomycin structure introduced into oritavancin and telavancin and five other vancomycin derivatives (with addition of 2-acetamido-2-deoxy-?-d-galactopyranosylamine, 2-acetamido-2-deoxy-?-d-glucopyranosylamine, 1-amine-1-deoxy-d-glucitol, 2-amino-2-deoxy-d-galactitol, or 2-amino-2-deoxy-d-glucitol to the C-terminal amino acid group in the vancomycin) are presented and discussed. The resulting molecular dynamics trajectories, root mean square deviation changes of aglycon and saccharide moieties as well as a comparative study of possible interactions with cyclic and chain forms of modified groups have been carried out, measured, and analyzed. Energetically advantageous conformations show close similarity to the structures known from the experimental data, but the diversity of others suggest very high conformational freedom of all modeled antibiotics and vancomycin derivatives. Alditol derivatives move closer to the peptidoglycan chain more easily but they also form intramolecular interactions more frequently than their homologous cyclic forms. One of the proposed derivatives seems to be a promising agent which is efficient in treatment of infections caused by Gram-positive bacteria. PMID:24685455

Slusarz, Rafa?; Szulc, Monika; Madaj, Janusz

2014-05-01

277

Sustained generation of electricity by the spore-forming, Gram-positive, Desulfitobacterium hafniense strain DCB2  

Microsoft Academic Search

Desulfitobacterium hafniense strain DCB2 generates electricity in microbial fuel cells (MFCs) when humic acids or the humate analog anthraquinone-2,6-disulfonate\\u000a (AQDS) is added as an electron-carrying mediator. When utilizing formate as fuel, the Gram-positive, spore-forming bacterium\\u000a generated up to 400 mW\\/m2 of cathode surface area in a single-chamber MFC with a platinum-containing air-fed cathode. Hydrogen, lactate, pyruvate,\\u000a and ethanol supported electricity generation,

C. E. Milliken; H. D. May

2007-01-01

278

Bactericidal effects induced by laser irradiation and haematoporphyrin against gram-positive and gram-negative microorganisms.  

PubMed

Laser irradiation of tissues treated in vivo with haematoporphyrin (Hpr) is known to result in a cytocidal effect, reportedly more pronounced in tumour tissues. To ascertain whether this cytocidal phenomenon can occur not only in eukaryotic but also in prokaryotic cells, the authors devised a model system in vitro consisting of bacterial cultures in liquid and in solid media. Bacteria photosensitized with Hpr were subsequently exposed to laser beams and to daylight; then normal microbiological techniques were used to determine whether any bactericidal effect occurred. Satisfactory results were achieved, particularly against Gram-positive microorganisms; a partial inhibition of Gram-negative microorganisms was also observed. PMID:3522156

Martinetto, P; Gariglio, M; Lombard, G F; Fiscella, B; Boggio, F

1986-01-01

279

Discriminatory antibacterial effects of calix[n]arene capped silver nanoparticles with regard to gram positive and gram negative bacteria.  

PubMed

Silver nanoparticles capped with nine different sulphonated calix[n]arenes were tested for their anti-bacterial effects against B. subtilis and E. coli at an apparent concentration of 100 nM in calix[n]arene. The results show the para-sulphonato-calix[n]arenes are active against Gram positive bacteria and the derivatives having sulphonate groups at both para and alkyl terminal positions are active against Gram negative bacteria. The calix[6]arene derivative with only O-alkyl sulphonate groups shows bactericidal activity. PMID:23831853

Boudebbouze, Samira; Coleman, Anthony W; Tauran, Yannick; Mkaouar, Hela; Perret, Florent; Garnier, Alexandrine; Brioude, Arnaud; Kim, Beomjoon; Maguin, Emmanuelle; Rhimi, Moez

2013-08-18

280

In Vitro Antimicrobial Activities of Novel Anilinouracils Which Selectively Inhibit DNA Polymerase III of Gram-Positive Bacteria  

Microsoft Academic Search

The 6-anilinouracils are novel dGTP analogs that selectively inhibit the replication-specific DNA polymerase III of gram-positive eubacteria. Two specific derivatives, IMAU (6-(3*-iodo-4*-methylanilino)uracil) and EMAU (6-(3*-ethyl-4*-methylanilino)uracil), were substituted with either a hydroxybutyl (HB) or a methoxy- butyl (MB) group at their N3 positions to produce four agents: HB-EMAU, MB-EMAU, HB-IMAU, and MB-IMAU. These four new agents inhibited Staphylococcus aureus, coagulase-negative staphylococci,

JENNIFER S. DALY; THEODORE J. GIEHL; NEAL C. BROWN; CHENGXIN ZHI; GEORGE E. WRIGHT; RICHARD T. ELLISON III

2000-01-01

281

Detection of Bacillus anthracis spores: comparison of quantum dot and organic dye labeling agents  

Microsoft Academic Search

Organic dyes are routinely used in labeling assays and sensing of biological molecules. Semiconductor quantum dots (QDs) have attractive features, particularly good resistance to photobleaching and narrow emission bands, which make them potential replacements for organic dyes. Using a previously identified synthetic peptide, a QD, and the R-phycoerythrin (RPE) dye, we have examined various labeling strategies for detecting Bacillus anthracis

William C. Schumacher; Andrew J. Phipps; Prabir K. Dutta

2009-01-01

282

STUDIES ON A BACTERICIDAL AGENT EXTRACTED FROM A SOIL BACILLUS  

PubMed Central

A Gram-positive, spore-bearing, aerobic bacillus, capable of lyzing the living cells of many Gram-positive microbial species, has been isolated from soil. Cultures of this soil bacillus in peptone media release during autolysis a soluble agent which exerts a bactericidal effect on all the Gram-positive microorganisms so far tested, and inactivates their glucose dehydrogenases. It also inhibits the growth of the susceptible species in culture media. Several of the Gram-positive species undergo lysis when incubated with the bactericidal agent. It appears however, that lysis is only a secondary process, due to the autolytic enzymes of the susceptible cells, and that it follows upon some other primary injury caused by the active agent. The bactericidal agent is ineffective against all the Gram-negative bacilli so far tested.

Dubos, Rene J.

1939-01-01

283

Identification and characterization of UDP-N-acetylenolpyruvylglucosamine reductase (MurB) from the Gram-positive pathogen Streptococcus pneumoniae.  

PubMed Central

The UDP-N-acetylenolpyruvylglucosamine reductase (MurB) from a Gram-positive pathogen, Streptococcus pneumoniae, was identified and characterized. The enzyme from S. pneumoniae shows 31% identity with the MurB protein from Escherichia coli, and contains the catalytic residues, substrate-binding residues and FAD-binding motif identified previously in the E. coli protein. The gene was cloned into the pET28a+ expression vector, and the 34.5 kDa protein that it encodes was overexpressed in E. coli strain BL21(DE3) to 30% of total cell protein. The majority of the protein was found to be insoluble. A variety of methods were used to increase the amount of soluble protein to 10%. This was then purified to near homogeneity in a two-step process. The absorption spectrum of the purified protein indicated it to be a flavoprotein, like its E. coli homologue, with a characteristic absorption at 463 nm. The enzyme was shown to be active, reducing UDP-N-acetylglucosamine enolpyruvate with the concomitant oxidation of NADPH, and was characterized kinetically with respect to its two substrates. The enzyme showed properties similar to those of its E. coli counterpart, being activated by univalent cations and being subject to substrate inhibition. The characterization of an important cell wall biosynthesis enzyme from a Gram-positive pathogen provides a good starting point for the discovery of antibacterial agents against MurB.

Sylvester, D R; Alvarez, E; Patel, A; Ratnam, K; Kallender, H; Wallis, N G

2001-01-01

284

A super-family of transcriptional activators regulates bacteriophage packaging and lysis in Gram-positive bacteria  

PubMed Central

The propagation of bacteriophages and other mobile genetic elements requires exploitation of the phage mechanisms involved in virion assembly and DNA packaging. Here, we identified and characterized four different families of phage-encoded proteins that function as activators required for transcription of the late operons (morphogenetic and lysis genes) in a large group of phages infecting Gram-positive bacteria. These regulators constitute a super-family of proteins, here named late transcriptional regulators (Ltr), which share common structural, biochemical and functional characteristics and are unique to this group of phages. They are all small basic proteins, encoded by genes present at the end of the early gene cluster in their respective phage genomes and expressed under cI repressor control. To control expression of the late operon, the Ltr proteins bind to a DNA repeat region situated upstream of the terS gene, activating its transcription. This involves the C-terminal part of the Ltr proteins, which control specificity for the DNA repeat region. Finally, we show that the Ltr proteins are the only phage-encoded proteins required for the activation of the packaging and lysis modules. In summary, we provide evidence that phage packaging and lysis is a conserved mechanism in Siphoviridae infecting a wide variety of Gram-positive bacteria.

Quiles-Puchalt, Nuria; Tormo-Mas, Maria Angeles; Campoy, Susana; Toledo-Arana, Alejandro; Monedero, Vicente; Lasa, Inigo; Novick, Richard P.; Christie, Gail E.; Penades, Jose R.

2013-01-01

285

A super-family of transcriptional activators regulates bacteriophage packaging and lysis in Gram-positive bacteria.  

PubMed

The propagation of bacteriophages and other mobile genetic elements requires exploitation of the phage mechanisms involved in virion assembly and DNA packaging. Here, we identified and characterized four different families of phage-encoded proteins that function as activators required for transcription of the late operons (morphogenetic and lysis genes) in a large group of phages infecting Gram-positive bacteria. These regulators constitute a super-family of proteins, here named late transcriptional regulators (Ltr), which share common structural, biochemical and functional characteristics and are unique to this group of phages. They are all small basic proteins, encoded by genes present at the end of the early gene cluster in their respective phage genomes and expressed under cI repressor control. To control expression of the late operon, the Ltr proteins bind to a DNA repeat region situated upstream of the terS gene, activating its transcription. This involves the C-terminal part of the Ltr proteins, which control specificity for the DNA repeat region. Finally, we show that the Ltr proteins are the only phage-encoded proteins required for the activation of the packaging and lysis modules. In summary, we provide evidence that phage packaging and lysis is a conserved mechanism in Siphoviridae infecting a wide variety of Gram-positive bacteria. PMID:23771138

Quiles-Puchalt, Nuria; Tormo-Más, María Ángeles; Campoy, Susana; Toledo-Arana, Alejandro; Monedero, Vicente; Lasa, Iñigo; Novick, Richard P; Christie, Gail E; Penadés, José R

2013-08-01

286

Gram-negative and Gram-Positive Bacterial Infections Give Rise to a Different Metabolic Response in a Mouse Model  

PubMed Central

Metabolomics has become an important tool to study host-pathogen interactions and to discover potential novel therapeutic targets. In an attempt to develop a better understanding of the process of pathogenesis and the associated host response we have used a quantitative 1H NMR approach to study the metabolic response to different bacterial infections. Here we describe that metabolic changes found in serum of mice that were infected with Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli and Pseudomonas aeruginosa can distinguish between infections caused by Gram-positive and Gram-negative bacterial strains. By combining the results of the mouse study with those of bacterial footprinting culture experiments, bacterially secreted metabolites could be identified as potential bacterium-specific biomarkers for P. aeruginosa infections but not for the other strains. Multivariate statistical analysis revealed correlations between metabolic, cytokine and physiological responses. In TLR4 and TLR2 knockout mice, host-response pathway correlated metabolites could be identified and allowed us for the first time to distinguish between bacterial- and host-induced metabolic changes. Since Gram-positive and Gram-negative bacteria activate different receptor pathways in the host, our results suggest that it may become possible in the future to use a metabolomics approach to improve on current clinical microbiology diagnostic methods.

2012-01-01

287

Targeting agr- and agr-Like Quorum Sensing Systems for Development of Common Therapeutics to Treat Multiple Gram-Positive Bacterial Infections  

PubMed Central

Invasive infection by the Gram-positive pathogen Staphylococcus aureus is controlled by a four gene operon, agr that encodes a quorum sensing system for the regulation of virulence. While agr has been well studied in S. aureus, the contribution of agr homologues and analogues in other Gram-positive pathogens is just beginning to be understood. Intriguingly, other significant human pathogens, including Clostridium perfringens, Listeria monocytogenes, and Enterococcus faecalis contain agr or analogues linked to virulence. Moreover, other significant human Gram-positive pathogens use peptide based quorum sensing systems to establish or maintain infection. The potential for commonality in aspects of these signaling systems across different species raises the prospect of identifying therapeutics that could target multiple pathogens. Here, we review the status of research into these agr homologues, analogues, and other peptide based quorum sensing systems in Gram-positive pathogens as well as the potential for identifying common pathways and signaling mechanisms for therapeutic discovery.

Gray, Brian; Hall, Pamela; Gresham, Hattie

2013-01-01

288

Bacillus cereus and related species.  

PubMed Central

Bacillus cereus is a gram-positive aerobic or facultatively anaerobic spore-forming rod. It is a cause of food poisoning, which is frequently associated with the consumption of rice-based dishes. The organism produces an emetic or diarrheal syndrome induced by an emetic toxin and enterotoxin, respectively. Other toxins are produced during growth, including phospholipases, proteases, and hemolysins, one of which, cereolysin, is a thiol-activated hemolysin. These toxins may contribute to the pathogenicity of B. cereus in nongastrointestinal disease. B. cereus isolated from clinical material other than feces or vomitus was commonly dismissed as a contaminant, but increasingly it is being recognized as a species with pathogenic potential. It is now recognized as an infrequent cause of serious nongastrointestinal infection, particularly in drug addicts, the immunosuppressed, neonates, and postsurgical patients, especially when prosthetic implants such as ventricular shunts are inserted. Ocular infections are the commonest types of severe infection, including endophthalmitis, panophthalmitis, and keratitis, usually with the characteristic formation of corneal ring abscesses. Even with prompt surgical and antimicrobial agent treatment, enucleation of the eye and blindness are common sequelae. Septicemia, meningitis, endocarditis, osteomyelitis, and surgical and traumatic wound infections are other manifestations of severe disease. B. cereus produces beta-lactamases, unlike Bacillus anthracis, and so is resistant to beta-lactam antibiotics; it is usually susceptible to treatment with clindamycin, vancomycin, gentamicin, chloramphenicol, and erythromycin. Simultaneous therapy via multiple routes may be required.

Drobniewski, F A

1993-01-01

289

Daptomycin Activity against Uncommonly Isolated Streptococcal and Other Gram-Positive Species Groups  

PubMed Central

A total of 1,356 clinical isolates were tested against daptomycin by broth microdilution methods. Daptomycin was active against seven groups of viridans group streptococci (MIC50 and MIC90 values ranging from ?0.06 and ?0.06 ?g/ml [Streptococcus bovis and Streptococcus dysgalactiae] to 0.5 and 1 ?g/ml [Streptococcus mitis, Streptococcus oralis, and Streptococcus parasanguinis], respectively), beta-hemolytic streptococci serogroups C, F, and G (MIC50 and MIC90, ?0.06 to 0.25 and 0.12 to 0.25 ?g/ml, respectively), Corynebacterium spp. (MIC50 and MIC90, ?0.06 and 0.12 ?g/ml, respectively), and Micrococcus spp. (MIC50 and MIC90, ?0.06 and 0.25 ?g/ml, respectively). Listeria monocytogenes exhibited higher daptomycin MICs (MIC50 and MIC90, 2 and 4 ?g/ml, respectively) than other tested organisms.

Flamm, Robert K.; Farrell, David J.; Jones, Ronald N.

2013-01-01

290

In vitro activities of peptide deformylase inhibitors against gram-positive pathogens.  

PubMed

The activities of six peptide deformylase (PDF) inhibitors against 107 respiratory tract pathogens were studied and compared to those of ciprofloxacin and amoxicillin-clavulanate. Against Streptococcus pneumoniae, BB-83698 and BB-83815 were the most active PDF inhibitors (MIC at which 90% of the organisms tested were inhibited [MIC(90)], 0.25 microg/ml). Five of the agents showed similar activity against Moraxella catarrhalis (MIC(90), 0.12 microg/ml). All PDF inhibitors were less active against Haemophilus influenzae; BB-3497 was the most active agent (MIC(90), 2 microg/ml). Five agents were studied against Chlamydia spp. and showed activity similar to that of ciprofloxacin (MIC, 0.5 to 4 microg/ml). This study demonstrates that PDF inhibitors have the potential to be developed for the treatment of respiratory tract infections. PMID:11897602

Wise, R; Andrews, J M; Ashby, J

2002-04-01

291

Daptomycin activity against uncommonly isolated streptococcal and other gram-positive species groups.  

PubMed

A total of 1,356 clinical isolates were tested against daptomycin by broth microdilution methods. Daptomycin was active against seven groups of viridans group streptococci (MIC50 and MIC90 values ranging from ?0.06 and ?0.06 ?g/ml [Streptococcus bovis and Streptococcus dysgalactiae] to 0.5 and 1 ?g/ml [Streptococcus mitis, Streptococcus oralis, and Streptococcus parasanguinis], respectively), beta-hemolytic streptococci serogroups C, F, and G (MIC50 and MIC90, ?0.06 to 0.25 and 0.12 to 0.25 ?g/ml, respectively), Corynebacterium spp. (MIC50 and MIC90, ?0.06 and 0.12 ?g/ml, respectively), and Micrococcus spp. (MIC50 and MIC90, ?0.06 and 0.25 ?g/ml, respectively). Listeria monocytogenes exhibited higher daptomycin MICs (MIC50 and MIC90, 2 and 4 ?g/ml, respectively) than other tested organisms. PMID:24080651

Sader, Helio S; Flamm, Robert K; Farrell, David J; Jones, Ronald N

2013-12-01

292

Isolation and Characterization of Four Gram-PositiveNickel-Tolerant Microorganisms from Contaminated Riparian Sediments  

SciTech Connect

Microbial communities from riparian sediments contaminatedwith high levels of Ni and U were examined for metal-tolerantmicroorganisms. Isolation of four aerobic Ni-tolerant, Gram-positiveheterotrophic bacteria indicated selection pressure from Ni. Theseisolates were identified as Arthrobacter oxydans NR-1, Streptomycesgalbus NR-2, Streptomyces aureofaciens NR-3, and Kitasatosporacystarginea NR-4 based on partial 16S rDNA sequences. A functional genemicroarray containing gene probes for functions associated withbiogeochemical cycling, metal homeostasis, and organic contaminantdegradation showed little overlap among the four isolates. Fifteen of thegenes were detected in all four isolates with only two of these relatedto metal resistance, specifically to tellurium. Each of the four isolatesalso displayed resistance to at least one of six antibiotics tested, withresistance to kanamycin, gentamycin, and ciprofloxacin observed in atleast two of the isolates. Further characterization of S. aureofaciensNR-3 and K. cystarginea NR-4 demonstrated that both isolates expressed Nitolerance constitutively. In addition, both were able to grow in higherconcentrations of Ni at pH 6 as compared to pH 7 (42.6 and 8.5 mM Ni atpH 6 and 7, respectively). Tolerance to Cd, Co, and Zn was also examinedin these two isolates; a similar pH-dependent metal tolerance wasobserved when grown with Co and Zn. Neither isolate was tolerant to Cd.These findings suggest that Ni is exerting a selection pressure at thissite for metal-resistant actinomycetes.

Van Nostrand, Joy D.; Khijniak, Tatiana V.; Gentry, Terry J.; Novak, Michelle T.; Sowder, Andrew G.; Zhou, Jizhong Z.; Bertsch, PaulM.; Morris, Pamela J.

2006-08-30

293

Production of equol from daidzein by gram-positive rod-shaped bacterium isolated from rat intestine.  

PubMed

Isoflavones (mainly daidzein and genistin) belong to the flavonoid group of compounds and are classified as phytoestrogens. In the intestine, daidzin is converted to daidzein by beta-glucosidase, and then daidzein is converted to O-desmethylangolensin (O-DMA) or equol via dihydrodaidzein by enzymes of intestinal bacteria. We isolated, for the first time, an anaerobic gram-positive rod-shaped strain capable of producing equol from daidzein. Its 16S rDNA gene sequence (1428 bp) showed 99% similarity with that of the human intestinal bacterium SNU-Julong 732 (AY310748) and 93% similarity with that of Eggerthella lenta ATCC 25559(T) (AF292375). This strain converted daidzein to equol via dihydrodaidzein in an equol-assay medium anaerobically. The addition of butyric acid and arginine increased the conversion ratio of daidzein to equol 4.7- and 4.5-fold, respectively. PMID:17046543

Minamida, Kimiko; Tanaka, Michiko; Abe, Ayumi; Sone, Teruo; Tomita, Fusao; Hara, Hiroshi; Asano, Kozo

2006-09-01

294

Plants used in Guatemala for the treatment of respiratory diseases. 1. Screening of 68 plants against gram-positive bacteria.  

PubMed

Respiratory ailments are important causes of morbidity and mortality in developing countries. Ethnobotanical surveys and literature reviews conducted in Guatemala during 1986-88 showed that 234 plants from 75 families, most of them of American origin, have been used for the treatment of respiratory ailments. Three Gram-positive bacteria causing respiratory infections (Staphylococcus aureus, Streptococcus pneumoniae and Streptococcus pyogenes) were used to screen 68 of the most commonly used plants for activity. Twenty-eight of these (41.2%) inhibited the growth of one or more of the bacteria tested. Staphylococcus aureus was inhibited by 18 of the plant extracts, while 7 extracts were effective against Streptococcus pyogenes. Plants of American origin which exhibited antibacterial activity were: Gnaphalium viscosum, Lippia alba, Lippia dulcis, Physalis philadelphica, Satureja brownei, Solanum nigrescens and Tagetes lucida. These preliminary in vitro results provide scientific basis for the use of these plants against bacterial respiratory infections. PMID:2023428

Caceres, A; Alvarez, A V; Ovando, A E; Samayoa, B E

1991-02-01

295

Functionalized magnetic iron oxide (Fe3O4) nanoparticles for capturing gram-positive and gram-negative bacteria.  

PubMed

The development of nanotechnology in biology and medicine has raised the need for conjugation of nanoparticles (NPs) to biomolecules. In this study, magnetic and functionalized magnetic iron oxide nanoparticles were synthesized and used as affinity probes to capture Gram-positive/negative bacteria. The morphology and properties of the magnetic NPs were examined by transmission electron microscopy, Fourier transform infrared spectroscopy, and zeta potential measurements. Furthermore, this study investigated the interaction between functionalized magnetic nanoparticles and Gram positive/negative bacteria. The positively and negatively charged magnetic nanoparticles include functionalities of Fe3O4, SiO2, TiO2, ZrO2, poly ethyleneimine (PEI) and poly acrylic acid. Their capture efficiencies for bacteria were investigated based on factors such as zeta potential, concentration and pH value. PEI particles carry a positive charge over a range of pH values from 3 to 10, and the particles were found to be an excellent candidate for capturing bacteria over such pH range. Since the binding force is mainly electrostatic, the architecture and orientation of the functional groups on the NP surface are not critical. Finally the captured bacteria were analyzed using matrix-assisted laser desorption/ionization mass spectrometry. The minimum detection limit was 10(4) CFU/mL and the analysis time was reduced to be less than 1 hour. In addition, the detection limit could be reduced to an extremely low concentration of 50 CFU/mL when captured bacteria were cultivated. PMID:25016643

Reddy, P Muralidhar; Chang, Kai-Chih; Liu, Zhen-Jun; Chen, Cheng-Tung; Ho, Yen-Peng

2014-08-01

296

Detection of Human Intestinal Catalase-Negative, Gram-Positive Cocci by rRNA-Targeted Reverse Transcription-PCR? †  

PubMed Central

An analytical system based on rRNA-targeted reverse transcription-quantitative PCR (RT-qPCR) for enumeration of catalase-negative, Gram-positive cocci was established. Subgroup- or species-specific primer sets targeting 16S or 23S rRNA from Enterococcus, Streptococcus, and Lactococcus were newly developed. The RT-qPCR method using these primers together with the previously reported primer sets specific for the Enterococcus genus, the Streptococcus genus, and several Streptococcus species was found to be able to quantify the target populations with detection limits of 103 to 104 cells per gram feces, which was more than 100 times as sensitive as the qPCR method (106 to 108 cells per gram feces). The RT-qPCR analysis of fecal samples from 24 healthy adult volunteers using the genus-specific primer sets revealed that Enterococcus and Streptococcus were present as intestinal commensals at population levels of log10 6.2 ± 1.4 and 7.5 ± 0.9 per gram feces (mean ± standard deviation [SD]), respectively. Detailed investigation using species- or subgroup-specific primer sets revealed that the volunteers harbored unique Enterococcus species, including the E. avium subgroup, the E. faecium subgroup, E. faecalis, the E. casseliflavus subgroup, and E. caccae, while the dominant human intestinal Streptococcus species was found to be S. salivarius. Various Lactococcus species, such as L. lactis subsp. lactis or L. lactis subsp. cremoris, L. garvieae, L. piscium, and L. plantarum, were also detected but at a lower population level (log10 4.6 ± 1.2 per gram feces) and prevalence (33%). These results suggest that the RT-qPCR method enables the accurate and sensitive enumeration of human intestinal subdominant but still important populations, such as Gram-positive cocci.

Kubota, Hiroyuki; Tsuji, Hirokazu; Matsuda, Kazunori; Kurakawa, Takashi; Asahara, Takashi; Nomoto, Koji

2010-01-01

297

Differential mode of antimicrobial actions of arginine-rich and lysine-rich histones against Gram-positive Staphylococcus aureus.  

PubMed

We previously reported the activities and modes of action of arginine (Arg)-rich histones H3 and H4 against Gram-negative bacteria. In the present study, we investigated the properties of the Arg-rich histones against Gram-positive bacteria in comparison with those of lysine (Lys)-rich histone H2B. In a standard microdilution assay, calf thymus histones H2B, H3, and H4 showed growth inhibitory activity against Staphylococcus aureus with minimum effective concentration values of 4.0, 4.0, and 5.6 ?M, respectively. Laser confocal microscopic analyses revealed that both the Arg-rich and Lys-rich histones associated with the surface of S. aureus. However, while the morphology of S. aureus treated with histone H2B appeared intact, those treated with the histones H3 and H4 closely resembled each other, and the cells were blurred. Electrophoretic mobility shift assay results revealed these histones have binding affinity to lipoteichoic acid (LTA), one of major cell surface components of Gram-positive bacteria. Scanning electron microscopic analyses demonstrated that while histone H2B elicited no obvious changes in cell morphology, histones H3 and H4 disrupted the cell membrane structure with bleb formation in a manner similar to general antimicrobial peptides. Consequently, our results suggest that bacterial cell surface LTA initially attracts both the Arg- and Lys-rich histones, but the modes of antimicrobial action of these histones are different; the former involves cell membrane disruption and the latter involves the cell integrity disruption. PMID:23932939

Morita, Shuu; Tagai, Chihiro; Shiraishi, Takayuki; Miyaji, Kazuyuki; Iwamuro, Shawichi

2013-10-01

298

Size-dependent antimicrobial properties of CuO nanoparticles against Gram-positive and -negative bacterial strains  

PubMed Central

Background CuO is one of the most important transition metal oxides due to its captivating properties. It is used in various technological applications such as high critical temperature superconductors, gas sensors, in photoconductive applications, and so on. Recently, it has been used as an antimicrobial agent against various bacterial species. Here we synthesized different sized CuO nanoparticles and explored the size-dependent antibacterial activity of each CuO nanoparticles preparation. Methods CuO nanoparticles were synthesized using a gel combustion method. In this approach, cupric nitrate trihydrate and citric acid were dissolved in distilled water with a molar ratio of 1:1. The resulting solution was stirred at 100°C, until gel was formed. The gel was allowed to burn at 200°C to obtain amorphous powder, which was further annealed at different temperatures to obtain different size CuO nanoparticles. We then tested the antibacterial properties using well diffusion, minimum inhibitory concentration, and minimum bactericidal concentration methods. Results XRD spectra confirmed the formation of single phase CuO nanoparticles. Crystallite size was found to increase with an increase in annealing temperature due to atomic diffusion. A minimum crystallite size of 20 nm was observed in the case of CuO nanoparticles annealed at 400°C. Transmission electron microscopy results corroborate well with XRD results. All CuO nanoparticles exhibited inhibitory effects against both Gram-positive and -negative bacteria. The size of the particles was correlated with its antibacterial activity. Conclusion The antibacterial activity of CuO nanoparticles was found to be size-dependent. In addition, the highly stable minimum-sized monodispersed copper oxide nanoparticles synthesized during this study demonstrated a significant increase in antibacterial activities against both Gram-positive and -negative bacterial strains.

Azam, Ameer; Ahmed, Arham S; Oves, M; Khan, MS; Memic, Adnan

2012-01-01

299

Molecular, technological and safety characterization of Gram-positive catalase-positive cocci from slightly fermented sausages.  

PubMed

The population of Gram-positive catalase-positive cocci from slightly fermented sausages was characterized at species and strain level by molecular techniques and some technological and hygienic aspects were also considered. Staphylococcus xylosus was the predominant species (80.8%) followed by Staphylococcus warneri (8.3%), Staphylococcus epidermidis (5.8%) Staphylococcus carnosus (4.6%), and Kocuria varians (0.4%). Proteolytic activity was observed in 23% of the isolates. The species with the highest percentage of proteolytic strains was S. warneri. Lipolytic activity was found in 45.8% of the isolates and S. xylosus was the species with the highest percentage of lipolytic isolates. Biogenic amine production was not widely distributed (only 14.6% of the isolates). Tyramine was the most intense amine produced, although by only 4.6% of the isolates. Phenylethylamine was more frequently detected (10.8% of isolates) but at lower levels. Some strains also produced putrescine (3.3%), cadaverine (2.9%), histamine (1.3%) and tryptamine (0.4%). All isolates were susceptible to linezolid and vancomicin and over 70% were resistant to penicillin G, ampicillin and sulphonamides. Most of the mecA+ strains (only 4.6% of isolates) also displayed resistance to multiple antibiotics. A reduced enterotoxigenic potential was found. Only 3.3% of isolates showed staphylococcal enterotoxins genes, all identified as entC gene. The combination of RAPD-PCR and plasmid profiling allowed the discrimination of 208 different profiles among the 240 Gram-positive catalase-positive cocci characterized, indicating a great genetic variability. PMID:16297478

Martín, B; Garriga, M; Hugas, M; Bover-Cid, S; Veciana-Nogués, M T; Aymerich, T

2006-03-15

300

[The cytomorphological evaluation of the effect of Bacillus intermedius RNAse on the organs of experimental animals].  

PubMed

Cytomorphological changes in organs of experimental animals after RNAse Bacillus intermedius treatment have been studied. It has been found that RNAse stimulates elements of lympho- and homopies of thymus and spleen and increases stromal reaction and functional activity of liver. The caused effects don't depend on catalytic enzyme activity. PMID:1302511

Nekhoroshkova, Z M; Kurinenko, B M; Khusnullina, K T; Karpova, S I; Bulgakova, R Sh

1992-01-01

301

Draft genome sequence of Bacillus oceanisediminis 2691.  

PubMed

Bacillus oceanisediminis 2691 is an aerobic, Gram-positive, spore-forming, and moderately halophilic bacterium that was isolated from marine sediment of the Yellow Sea coast of South Korea. Here, we report the draft genome sequence of B. oceanisediminis 2691 that may have an important role in the bioremediation of marine sediment. PMID:23105082

Lee, Yong-Jik; Lee, Sang-Jae; Jeong, Haeyoung; Kim, Hyun Ju; Ryu, Naeun; Kim, Byoung-Chan; Lee, Han-Seung; Lee, Dong-Woo; Lee, Sang Jun

2012-11-01

302

Draft Genome Sequence of Bacillus oceanisediminis 2691  

PubMed Central

Bacillus oceanisediminis 2691 is an aerobic, Gram-positive, spore-forming, and moderately halophilic bacterium that was isolated from marine sediment of the Yellow Sea coast of South Korea. Here, we report the draft genome sequence of B. oceanisediminis 2691 that may have an important role in the bioremediation of marine sediment.

Lee, Yong-Jik; Lee, Sang-Jae; Jeong, Haeyoung; Kim, Hyun Ju; Ryu, Naeun; Kim, Byoung-Chan; Lee, Han-Seung

2012-01-01

303

Complete Genome of Bacillus thuringiensis Myophage Spock  

PubMed Central

Bacillus thuringiensis is a Gram-positive, sporulating soil microbe with valuable pesticide-producing properties. The study of bacteriophages of B. thuringiensis could provide new biotechnological tools for the use of this bacterium. Here, we present the complete annotated genome of Spock, a myophage of B. thuringiensis, and describe its features.

Maroun, Justin W.; Whitcher, Kelvin J.; Chamakura, Karthik R.

2013-01-01

304

In vitro activity of tigecycline (GAR-936) tested against 11,859 recent clinical isolates associated with community-acquired respiratory tract and gram-positive cutaneous infections.  

PubMed

Tigecycline is a novel 9-t-butylglycylamido derivative of minocycline that has demonstrated activity against a variety of bacterial pathogens, including resistant isolates, during preclinical studies. In vitro activities of tigecycline and comparators were tested against 11,859 recent (2000 and 2002) bacterial strains recovered from patients in 29 countries with community-acquired respiratory tract disease (3,317 gram-positive and -negative strains) and skin and soft tissue infections (8,542 gram-positive strains). All oxacillin-susceptible and -resistant Staphylococcus aureus (5,077 strains; tigecycline MIC(90), 0.5 microg/mL) and coagulase-negative staphylococci (1,432 strains; MIC(90), 0.5 microg/mL), penicillin-susceptible and -resistant Streptococcus pneumoniae (1,585 strains; MIC(90), < or =0.25 microg/mL), viridans group streptococci (212 strains; MIC(90), < or =0.25-0.5 microg/mL), vancomycin-susceptible and -resistant enterococci (1,416 strains; MIC(90), 0.25-0.5 microg/mL), beta-haemolytic streptococci (405 strains; MIC(90), < or =0.25 microg/mL), beta-lactamase positive and negative Haemophilus influenzae (1,220 strains; MIC(90), 1 microg/mL), Moraxella catarrhalis (495 strains; MIC(90), 0.25 microg/mL), and Neisseria meningitidis (17 strains; MIC(90), < or =0.12 microg/mL) were inhibited by 2 microg/mL or less of tigecycline. Whereas potency of tetracycline and doxycycline markedly dropped in various resistant organism subsets, tigecycline was unaffected with an overall MIC(90) of 0.5 microg/mL. These findings confirm that tigecycline maintains a truly broad spectrum like the tetracycline class while enhancing potency. It also incorporates stability to the commonly occurring tetracycline resistance mechanisms, making it an attractive candidate for continued clinical development against pathogens causing serious community-acquired respiratory tract infections, as well as cutaneous infections. PMID:15246511

Fritsche, Thomas R; Kirby, Jeffrey T; Jones, Ronald N

2004-07-01

305

Crystal Structure of 3-Chlorocatechol 1,2-dioxygenase Key Enzyme of a New Modified Ortho-pathway from the Gram-positive Rhodococcus opacus 1CP Grown on 2-chlorophenol  

Microsoft Academic Search

The crystal structure of the 3-chlorocatechol 1,2-dioxygenase from the Gram-positive bacterium Rhodococcus opacus (erythropolis) 1CP, a Fe(III) ion-containing enzyme specialized in the aerobic biodegradation of 3-chloro- and methyl-substituted catechols, has been solved by molecular replacement techniques using the coordinates of 4-chlorocatechol 1,2-dioxygenase from the same organism (PDB code 1S9A) as a starting model and refined at 1.9 Å resolution (Rfree 21.9%;

Marta Ferraroni; Marina P. Kolomytseva; Inna P. Solyanikova; Andrea Scozzafava; Ludmila A. Golovleva; Fabrizio Briganti

2006-01-01

306

Phylogenetic analysis of Butyrivibrio strains reveals three distinct groups of species within the Clostridium subphylum of the gram-positive bacteria.  

PubMed

The phylogenetic positions of 40 Butyrivibrio strains were determined by performing a comparative sequence analysis of the 16S rRNA genes of these organisms. We found that all of the strains which we studied belong to cluster XIVa (M. D. Collins, P. A. Lawson, A. Willems, J. J. Cordoba, J. Fernandez=Garayzabal, P. Garcia, J. Cai, H. Hippe, and J. A. E. Farrow, Int. J. Syst. Bacteriol. 44:812-826, 1994) of the Clostridium subphylum of the gram-positive bacteria, which also includes several Clostridium, Coprococcus, Eubacterium, and Ruminococcus species. We also found that the Butyrivibrio strains which we examined were genotypically heterogeneous and exhibited 12 distinct rRNA sequence types. The 12 rRNA sequence types formed three distinct lineages in cluster XIVa, which were separate from each other and from all other species belonging to this cluster. One lineage consisted of strains which exhibited a single rRNA type and corresponded to the species Butyrivibrio crossotus. The second lineage consisted of 12 strains designated Butyrivibrio fibrisolvens which exhibited seven distinct rRNA sequence types. The type strain of B. fibrisolvens was a member of this lineage, but its position was peripheral. The third lineage comprised 26 B. fibrisolvens strains which exhibited four distinct rRNA sequence types. Tree topology and sequence divergence considerations indicated that the three lineages correspond to three separate genera and that the genus Butyrivibrio should be restricted to the group that contains the type strain of B. fibrisolvens. PMID:8573495

Willems, A; Amat-Marco, M; Collins, M D

1996-01-01

307

Transcriptional Organization and Posttranscriptional Regulation of the Bacillus subtilis BranchedChain Amino Acid Biosynthesis Genes  

Microsoft Academic Search

In Bacillus subtilis, the genes of the branched-chain amino acids biosynthetic pathway are organized in three genetic loci: the ilvBHC-leuABCD (ilv-leu) operon, ilvA, and ilvD. These genes, as well as ybgE, encoding a branched-chain amino acid aminotransferase, were recently demonstrated to represent direct targets of the global transcriptional regulator CodY. In the present study, the transcriptional organization and posttran- scriptional

Ulrike Mader; Susanne Hennig; Michael Hecker; Georg Homuth

2004-01-01

308

Efficacy of the new lantibiotic NAI-107 in experimental infections induced by multidrug-resistant Gram-positive pathogens.  

PubMed

NAI-107 is a novel lantibiotic active against Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA), glycopeptide-intermediate S. aureus (GISA), and vancomycin-resistant enterococci (VRE). The aim of this study was to evaluate the in vivo efficacy of NAI-107 in animal models of severe infection. In acute lethal infections induced with a penicillin-intermediate Streptococcus pneumoniae strain in immunocompetent mice, or with MRSA, GISA, and VRE strains in neutropenic mice, the 50% effective dose (ED(50)) values of NAI-107 were comparable or lower than those of reference compounds, irrespective of the strain and immune status (0.51 to 14.2 mg/kg of body weight for intravenous [i.v.] NAI-107, 5.1 to 22.4 for oral linezolid, and 22.4 for subcutaneous [s.c.] vancomycin). In the granuloma pouch model induced in rats with a MRSA strain, intravenous NAI-107 showed a dose-proportional bactericidal activity that, at a single 40-mg/kg dose, compared with 2 20-mg/kg doses at a 12-h or 24-h interval, caused a 3-log(10)-CFU/ml reduction of viable MRSA in exudates that persisted for more than 72 h. Rat endocarditis was induced with a MRSA strain and treated for five consecutive days. In a first experiment, using 5, 10, or 20 mg/kg/day, and in a second experiment, when 10 mg/kg at 12-h intervals was compared to 20 mg/kg/day, intravenous NAI-107 was effective in reducing the bacterial load in heart vegetations in a dose-proportional manner. Trough plasma levels, as determined on days 2 and 5, were several times higher than the NAI-107 minimal bactericidal concentration (MBC). NAI-107 binding to rat and human serum ranges between 93% and 98.6%. The rapid bactericidal activity of NAI-107 observed in vitro was thus confirmed by the efficacy in several models of experimental infection induced by Gram-positive pathogens, supporting further investigation of the compound. PMID:21220527

Jabés, Daniela; Brunati, Cristina; Candiani, GianPaolo; Riva, Simona; Romanó, Gabriella; Donadio, Stefano

2011-04-01

309

Unravelling a vicious circle: animal feed marketed in Costa Rica contains irregular concentrations of tetracyclines and abundant oxytetracycline-resistant Gram-positive bacteria.  

PubMed

Diverse tetracyclines are used to prevent and control bacterial infections in livestock and farmed fish. These drugs are administered through the diet, but farmers seldom check whether feed contains antibiotic-resistant bacteria that may colonise their crops or transfer their resistance traits to species of veterinary relevance. To examine whether antibiotic dosage defines the abundance of antibiotic-resistant bacteria in animal feed, we determined the concentration of parental compounds and epimers of oxytetracycline (OTC), doxycycline, tetracycline and chlortetracycline, as well as the abundance and resistance level of OTC-resistant bacteria in samples of fish (n = 21), poultry (n = 21), swine (n = 21), and shrimp feed (n = 21) marketed in Costa Rica. Fish feed contained the highest amounts of tetracyclines (119-8365 mg kg(-1)) and the largest proportion of bacteria resistant to 10 ?g ml(-1) (1.8-92.4%) or 100 ?g ml(-1) of OTC (12.5-63.8%). Poultry (78-438 mg kg(-1)) and swine (41-1076 mg kg(-1)) feed had intermediate concentrations of tetracyclines and OTC-resistant bacteria (0.2-66% and 0.3-49%, respectively), whereas shrimp feed showed the lowest amounts of tetracyclines (21.5-50.3 mg kg(-1)), no OTC and no culturable OTC-resistant bacteria. In line with these results, the MIC50 of OTC for 150 isolates from fish and poultry feed was > 256 µg ml(-1), while that of 150 bacteria isolated from swine feed was 192 µg ml(-1). Phenotypic tests, fatty acid profiles and proteotypic analyses by matrix-assisted laser desorption/ionisation-time of flight mass-spectroscopy revealed that most OTC-resistant isolates were Gram-positive bacteria of low G+C% content from the genera Staphylococcus and Bacillus. Clear correlations between OTC dosage and feed colonisation with OTC-resistant bacteria were seen in medicated feed for fish (r = 0.179-0.651). Nonetheless, some unmedicated feed for fish, swine and poultry contained large populations of OTC-resistant bacteria, suggesting that raw materials and manufacturing processes may also influence carriage of OTC-resistant bacteria in animal feed. PMID:24660748

Granados-Chinchilla, Fabio; Alfaro, Margarita; Chavarría, Guadalupe; Rodríguez, César

2014-06-01

310

Lagging strand replication of rolling-circle plasmids: Specific recognition of the ssoA-type origins in different gram-positive bacteria  

PubMed Central

Many bacterial plasmids replicate by a rolling-circle mechanism that involves the generation of single-stranded DNA (ssDNA) intermediates. Replication of the lagging strand of such plasmids initiates from their single strand origin (sso). Many different types of ssos have been identified. One group of ssos, termed ssoA, which have conserved sequence and structural features, function efficiently only in their natural hosts in vivo. To study the host specificity of sso sequences, we have analyzed the functions of two closely related ssoAs belonging to the staphylococcal plasmid pE194 and the streptococcal plasmid pLS1 in Staphylococcus aureus. The pLS1 ssoA functioned poorly in vivo in S. aureus as evidenced by accumulation of high levels of ssDNA but supported efficient replication in vitro in staphylococcal extracts. These results suggest that one or more host factors that are present in sufficient quantities in S. aureus cell-free extracts may be limiting in vivo. Mapping of the initiation points of lagging strand synthesis in vivo and in vitro showed that DNA synthesis initiates from specific sites within the pLS1 ssoA. These results demonstrate that specific initiation of replication can occur from the pLS1 ssoA in S. aureus although it plays a minimal role in lagging strand synthesis in vivo. Therefore, the poor functionality of the pLS1 in vivo in a nonnative host is caused by the low efficiency rather than a lack of specificity of the initiation process. We also have identified ssDNA promoters and mapped the primer RNAs synthesized by the S. aureus and Bacillus subtilis RNA polymerases from the pE194 and pLS1 ssoAs. The S. aureus RNA polymerase bound more efficiently to the native pE194 ssoA as compared with the pLS1 ssoA, suggesting that the strength of RNA polymerase–ssoA interaction may play a major role in the functionality of the ssoA sequences in Gram-positive bacteria.

Kramer, M. Gabriela; Espinosa, Manuel; Misra, Tapan K.; Khan, Saleem A.

1998-01-01

311

Bacillus anthracis genome organization in light of whole transcriptome sequencing  

SciTech Connect

Emerging knowledge of whole prokaryotic transcriptomes could validate a number of theoretical concepts introduced in the early days of genomics. What are the rules connecting gene expression levels with sequence determinants such as quantitative scores of promoters and terminators? Are translation efficiency measures, e.g. codon adaptation index and RBS score related to gene expression? We used the whole transcriptome shotgun sequencing of a bacterial pathogen Bacillus anthracis to assess correlation of gene expression level with promoter, terminator and RBS scores, codon adaptation index, as well as with a new measure of gene translational efficiency, average translation speed. We compared computational predictions of operon topologies with the transcript borders inferred from RNA-Seq reads. Transcriptome mapping may also improve existing gene annotation. Upon assessment of accuracy of current annotation of protein-coding genes in the B. anthracis genome we have shown that the transcriptome data indicate existence of more than a hundred genes missing in the annotation though predicted by an ab initio gene finder. Interestingly, we observed that many pseudogenes possess not only a sequence with detectable coding potential but also promoters that maintain transcriptional activity.

Martin, Jeffrey; Zhu, Wenhan; Passalacqua, Karla D.; Bergman, Nicholas; Borodovsky, Mark

2010-03-22

312

Identification of a ligand on the Wip1 bacteriophage highly specific for a receptor on Bacillus anthracis.  

PubMed

Tectiviridae is a family of tailless bacteriophages with Gram-negative and Gram-positive hosts. The family model PRD1 and its close relatives all infect a broad range of enterobacteria by recognizing a plasmid-encoded conjugal transfer complex as a receptor. In contrast, tectiviruses with Gram-positive hosts are highly specific to only a few hosts within the same bacterial species. The cellular determinants that account for the observed specificity remain unknown. Here we present the genome sequence of Wip1, a tectivirus that infects the pathogen Bacillus anthracis. The Wip1 genome is related to other tectiviruses with Gram-positive hosts, notably, AP50, but displays some interesting differences in its genome organization. We identified Wip1 candidate genes for the viral spike complex, the structure located at the capsid vertices and involved in host receptor binding. Phage adsorption and inhibition tests were combined with immunofluorescence microscopy to show that the Wip1 gene product p23 is a receptor binding protein. His-p23 also formed a stable complex with p24, a Wip1 protein of unknown function, suggesting that the latter is involved with p23 in host cell recognition. The narrow host range of phage Wip1 and the identification of p23 as a receptor binding protein offer a new range of suitable tools for the rapid identification of B. anthracis. PMID:23893110

Kan, Sherry; Fornelos, Nadine; Schuch, Raymond; Fischetti, Vincent A

2013-10-01

313

Identification of a Ligand on the Wip1 Bacteriophage Highly Specific for a Receptor on Bacillus anthracis  

PubMed Central

Tectiviridae is a family of tailless bacteriophages with Gram-negative and Gram-positive hosts. The family model PRD1 and its close relatives all infect a broad range of enterobacteria by recognizing a plasmid-encoded conjugal transfer complex as a receptor. In contrast, tectiviruses with Gram-positive hosts are highly specific to only a few hosts within the same bacterial species. The cellular determinants that account for the observed specificity remain unknown. Here we present the genome sequence of Wip1, a tectivirus that infects the pathogen Bacillus anthracis. The Wip1 genome is related to other tectiviruses with Gram-positive hosts, notably, AP50, but displays some interesting differences in its genome organization. We identified Wip1 candidate genes for the viral spike complex, the structure located at the capsid vertices and involved in host receptor binding. Phage adsorption and inhibition tests were combined with immunofluorescence microscopy to show that the Wip1 gene product p23 is a receptor binding protein. His-p23 also formed a stable complex with p24, a Wip1 protein of unknown function, suggesting that the latter is involved with p23 in host cell recognition. The narrow host range of phage Wip1 and the identification of p23 as a receptor binding protein offer a new range of suitable tools for the rapid identification of B. anthracis.

Kan, Sherry; Fornelos, Nadine; Schuch, Raymond

2013-01-01

314

Complete genome sequence of the industrial bacterium Bacillus licheniformis and comparisons with closely related Bacillus species  

PubMed Central

Background Bacillus licheniformis is a Gram-positive, spore-forming soil bacterium that is used in the biotechnology industry to manufacture enzymes, antibiotics, biochemicals and consumer products. This species is closely related to the well studied model organism Bacillus subtilis, and produces an assortment of extracellular enzymes that may contribute to nutrient cycling in nature. Results We determined the complete nucleotide sequence of the B. licheniformis ATCC 14580 genome which comprises a circular chromosome of 4,222,336 base-pairs (bp) containing 4,208 predicted protein-coding genes with an average size of 873 bp, seven rRNA operons, and 72 tRNA genes. The B. licheniformis chromosome contains large regions that are colinear with the genomes of B. subtilis and Bacillus halodurans, and approximately 80% of the predicted B. licheniformis coding sequences have B. subtilis orthologs. Conclusions Despite the unmistakable organizational similarities between the B. licheniformis and B. subtilis genomes, there are notable differences in the numbers and locations of prophages, transposable elements and a number of extracellular enzymes and secondary metabolic pathway operons that distinguish these species. Differences include a region of more than 80 kilobases (kb) that comprises a cluster of polyketide synthase genes and a second operon of 38 kb encoding plipastatin synthase enzymes that are absent in the B. licheniformis genome. The availability of a completed genome sequence for B. licheniformis should facilitate the design and construction of improved industrial strains and allow for comparative genomics and evolutionary studies within this group of Bacillaceae.

Rey, Michael W; Ramaiya, Preethi; Nelson, Beth A; Brody-Karpin, Shari D; Zaretsky, Elizabeth J; Tang, Maria; de Leon, Alfredo Lopez; Xiang, Henry; Gusti, Veronica; Clausen, Ib Groth; Olsen, Peter B; Rasmussen, Michael D; Andersen, Jens T; J?rgensen, Per L; Larsen, Thomas S; Sorokin, Alexei; Bolotin, Alexander; Lapidus, Alla; Galleron, Nathalie; Ehrlich, S Dusko; Berka, Randy M

2004-01-01

315

Sustained generation of electricity by the spore-forming, Gram-positive, Desulfitobacterium hafniense strain DCB2.  

PubMed

Desulfitobacterium hafniense strain DCB2 generates electricity in microbial fuel cells (MFCs) when humic acids or the humate analog anthraquinone-2,6-disulfonate (AQDS) is added as an electron-carrying mediator. When utilizing formate as fuel, the Gram-positive, spore-forming bacterium generated up to 400 mW/m2 of cathode surface area in a single-chamber MFC with a platinum-containing air-fed cathode. Hydrogen, lactate, pyruvate, and ethanol supported electricity generation, but acetate, propionate, and butyrate did not. Scanning electron microscopy indicated that strain DCB2 colonized the surface of a current-generating anode but not of an unconnected electrode. The electricity was recovered fully within minutes after the exchange of the medium in the anode chamber and within a week after an exposure of a colonized anode to 90 degrees C for 20 min. Of the six strains of Desulfitobacteria tested, all of which would reduce AQDS, only D. hafniense strain DCB2 continued to reduce AQDS and generate electricity for more than 24 h, indicating that reduction of the humate analog alone is insufficient to sustain electrode reduction. PMID:17031638

Milliken, C E; May, H D

2007-01-01

316

Presence and dissemination of the multiresistance gene cfr in Gram-positive and Gram-negative bacteria.  

PubMed

The emergence of the multiresistance gene cfr in staphylococci is of global concern. In addition to conferring resistance to phenicols, lincosamides, pleuromutilins, streptogramin A antibiotics and selected 16-membered macrolides, the cfr gene also confers resistance to the oxazolidinone linezolid. Linezolid is a last-resort antimicrobial agent for the treatment of serious infections in humans caused by resistant Gram-positive bacteria. The cfr gene is often located on plasmids and several cfr-carrying plasmids have been described, which differ in their structure, their size and the presence of additional resistance genes. These plasmids are important vehicles that promote the spread of the cfr gene not only among bacteria of the same species, but also among those of different species and genera. Moreover, the cfr gene has been identified in close proximity to different insertion sequences, which most probably also play an important role in its dissemination. This review summarizes current knowledge on the genetic environment of the multiresistance gene cfr with particular reference to mobile genetic elements and co-located resistance genes that may support its emergence. PMID:23543608

Shen, Jianzhong; Wang, Yang; Schwarz, Stefan

2013-08-01

317

PorA Represents the Major Cell Wall Channel of the Gram-Positive Bacterium Corynebacterium glutamicum  

PubMed Central

The cell wall of the gram-positive bacterium Corynebacterium glutamicum contains a channel (porin) for the passage of hydrophilic solutes. The channel-forming polypeptide PorA is a 45-amino-acid acidic polypeptide with an excess of four negatively charged amino acids, which is encoded by the 138-bp gene porA. porA was deleted from the chromosome of C.glutamicum wild-type strain ATCC 13032 to obtain mutant ATCC 13032?porA. Southern blot analysis demonstrated that porA was deleted. Lipid bilayer experiments revealed that PorA was not present in the cell wall of the mutant strain. Searches within the known chromosome of C. glutamicum by using National Center for Biotechnology Information BLAST and reverse transcription-PCR showed that no other PorA-like protein is encoded on the chromosome or is expressed in the deletion strain. The porA deletion strain exhibited slower growth and longer growth times than the C. glutamicum wild-type strain. Experiments with different antibiotics revealed that the susceptibility of the mutant strain was much lower than that of the wild-type C. glutamicum strain. The results presented here suggest that PorA represents a major hydrophilic pathway through the cell wall and that C. glutamicum contains cell wall channels which are not related to PorA.

Costa-Riu, Noelia; Burkovski, Andreas; Kramer, Reinhard; Benz, Roland

2003-01-01

318

The molecular switch that activates the cell wall anchoring step of pilus assembly in gram-positive bacteria.  

PubMed

Cell surface pili in gram-positive bacteria orchestrate the colonization of host tissues, evasion of immunity, and the development of biofilms. Recent work revealed that pilus assembly is a biphasic process wherein pilus polymerization is catalyzed by a pilus-specific sortase followed by cell wall anchoring of the pilus that is promoted by the housekeeping sortase. Here, we present molecular genetic and biochemical studies of a heterotrimeric pilus in Corynebacterium diphtheriae, uncovering the molecular switch that terminates pilus polymerization in favor of cell wall anchoring. The prototype pilus contains a major pilin (SpaA) forming the shaft, a tip pilin (SpaC), and another minor pilin (SpaB). Cells lacking SpaB form pilus fibers, but they are largely secreted in the medium, a phenotype also observed when cells lack the housekeeping sortase. Furthermore, the average pilus length is greatly increased in the absence of SpaB. Remarkably, a SpaB mutant that lacks the cell wall sorting signal but contains a critical lysine residue is incorporated in the pilus. However, the resulting pili fail to anchor to the cell wall. We propose that a specific minor pilin acts as the terminal subunit in pilus assembly. Cell wall anchoring ensues when the pilus polymer assembled on the pilus-specific sortase is transferred to the minor pilin presented by the housekeeping sortase via lysine-mediated transpeptidation. PMID:18779588

Mandlik, Anjali; Das, Asis; Ton-That, Hung

2008-09-16

319

Significantly higher procalcitonin levels could differentiate Gram-negative sepsis from Gram-positive and fungal sepsis.  

PubMed

Procalcitonin (PCT) levels can distinguish between infectious and non-infectious systemic inflammatory response. However, there are some differences between Gram-negative (G-), Gram-positive (G+), and fungal bloodstream infections, particularly in different cytokine profiles, severity and mortality. The aim of current study was to examine whether PCT levels can serve as a distinguishing mark between G+, G-, and fungal sepsis as well. One hundred and sixty-six septic patients with positive blood cultures were examined on C-reactive protein (CRP) and PCT on the same date of blood culture evaluation. The median (interquartile range, IQR) of CRP and PCT in G+, G-, and fungal cohorts and comparison of measured values between groups were made using the Kruskal-Wallis test with subsequent Bonferroni's corrections, with p < 0.05. In 83/166 (50 %) of blood cultures, G+ microbes, 78/166 (47 %) G- rods, and 5/166 (3 %) fungi were detected. PCT concentrations (ng/ml) were significantly higher in G- compared to other cohorts: 8.90 (1.88; 32.60) in G-, 0.73 (0.22; 3.40) in G+, and 0.58 (0.35; 0.73) in fungi (p < 0.00001). CRP concentrations did not differ significantly in groups. Significantly higher PCT levels could differentiate G- sepsis from G+ and fungemia. In contrast to CRP, PCT is a good discriminative biomarker in different bloodstream infections. PMID:22644264

Brodská, Helena; Malí?ková, Karin; Adámková, Václava; Benáková, Hana; Š?astná, Markéta Marková; Zima, Tomáš

2013-08-01

320

Antimicrobial photodynamic efficiency of novel cationic porphyrins towards periodontal Gram-positive and Gram-negative pathogenic bacteria.  

PubMed

The Gram-negative Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum are major causative agents of aggressive periodontal disease. Due to increase in the number of antibiotic-resistant bacteria, antimicrobial Photodynamic therapy (aPDT) seems to be a plausible alternative. In this work, photosensitization was performed on Gram-positive and Gram-negative bacteria in pure culture using new-age cationic porphyrins, namely mesoimidazolium-substituted porphyrin derivative (ImP) and pyridinium-substituted porphyrin derivative (PyP). The photophysical properties of both the sensitizers including absorption, fluorescence emission, quantum yields of the triplet excited states and singlet oxygen generation efficiencies were evaluated in the context of aPDT application. The studied porphyrins exhibited high ability to accumulate into bacterial cells with complete penetration into early stage biofilms. As compared with ImP, PyP was found to be more effective for photoinactivation of bacterial strains associated with periodontitis, without any signs of dark toxicity, owing to its high photocytotoxicity. PMID:24164211

Prasanth, Chandra Sekhar; Karunakaran, Suneesh C; Paul, Albish K; Kussovski, Vesselin; Mantareva, Vanya; Ramaiah, Danaboyina; Selvaraj, Leslie; Angelov, Ivan; Avramov, Latchezar; Nandakumar, Krishnankutty; Subhash, Narayanan

2014-01-01

321

Transcriptional profiling of Gram-positive Arthrobacter in the phyllosphere: induction of pollutant degradation genes by natural plant phenolic compounds.  

PubMed

Arthrobacter chlorophenolicus?A6 is a Gram-positive, 4-chlorophenol-degrading soil bacterium that was recently shown to be an effective colonizer of plant leaf surfaces. The genetic basis for this phyllosphere competency is unknown. In this paper, we describe the genome-wide expression profile of A.?chlorophenolicus on leaves of common bean (Phaseolus vulgaris) compared with growth on agar surfaces. In phyllosphere-grown cells, we found elevated expression of several genes known to contribute to epiphytic fitness, for example those involved in nutrient acquisition, attachment, stress response and horizontal gene transfer. A surprising result was the leaf-induced expression of a subset of the so-called cph genes for the degradation of 4-chlorophenol. This subset encodes the conversion of the phenolic compound hydroquinone to 3-oxoadipate, and was shown to be induced not only by 4-chlorophenol but also hydroquinone, its glycosylated derivative arbutin, and phenol. Small amounts of hydroquinone, but not arbutin or phenol, were detected in leaf surface washes of P.?vulgaris by gas chromatography-mass spectrometry. Our findings illustrate the utility of genomics approaches for exploration and improved understanding of a microbial habitat. Also, they highlight the potential for phyllosphere-based priming of bacteria to stimulate pollutant degradation, which holds promise for the application of phylloremediation. PMID:24373130

Scheublin, Tanja R; Deusch, Simon; Moreno-Forero, Silvia K; Müller, Jochen A; van der Meer, Jan Roelof; Leveau, Johan H J

2014-07-01

322

Differential Targeting of the E-Cadherin/?-Catenin Complex by Gram-Positive Probiotic Lactobacilli Improves Epithelial Barrier Function  

PubMed Central

The intestinal ecosystem is balanced by dynamic interactions between resident and incoming microbes, the gastrointestinal barrier, and the mucosal immune system. However, in the context of inflammatory bowel diseases (IBD), where the integrity of the gastrointestinal barrier is compromised, resident microbes contribute to the development and perpetuation of inflammation and disease. Probiotic bacteria have been shown to exert beneficial effects, e.g., enhancing epithelial barrier integrity. However, the mechanisms underlying these beneficial effects are only poorly understood. Here, we comparatively investigated the effects of four probiotic lactobacilli, namely, Lactobacillus acidophilus, L. fermentum, L. gasseri, and L. rhamnosus, in a T84 cell epithelial barrier model. Results of DNA microarray experiments indicating that lactobacilli modulate the regulation of genes encoding in particular adherence junction proteins such as E-cadherin and ?-catenin were confirmed by quantitative reverse transcription-PCR (qRT-PCR). Furthermore, we show that epithelial barrier function is modulated by Gram-positive probiotic lactobacilli via their effect on adherence junction protein expression and complex formation. In addition, incubation with lactobacilli differentially influences the phosphorylation of adherence junction proteins and the abundance of protein kinase C (PKC) isoforms such as PKC? that thereby positively modulates epithelial barrier function. Further insight into the underlying molecular mechanisms triggered by these probiotics might also foster the development of novel strategies for the treatment of gastrointestinal diseases (e.g., IBD).

Hummel, Stephanie; Veltman, Katharina; Cichon, Christoph; Sonnenborn, Ulrich

2012-01-01

323

Use of Enzyme Tests in Characterization and Identification of Aerobic and Facultatively Anaerobic Gram-Positive Cocci  

PubMed Central

The contribution of enzyme tests to the accurate and rapid routine identification of gram-positive cocci is introduced. The current taxonomy of the genera of aerobic and facultatively anaerobic cocci based on genotypic and phenotypic characterization is reviewed. The clinical and economic importance of members of these taxa is briefly summarized. Tables summarizing test schemes and kits available for the identification of staphylococci, enterococci, and streptococci on the basis of general requirements, number of tests, number of taxa, test classes, and completion times are discussed. Enzyme tests included in each scheme are compared on the basis of their synthetic moiety. The current understanding of the activity of enzymes important for classification and identification of the major groups, methods of testing, and relevance to the ease and speed of identification are reviewed. Publications describing the use of different identification kits are listed, and overall identification successes and problems are discussed. The relationships between the results of conventional biochemical and rapid enzyme tests are described and considered. The use of synthetic substrates for the detection of glycosidases and peptidases is reviewed, and the advantages of fluorogenic synthetic moieties are discussed. The relevance of enzyme tests to accurate and meaningful rapid routine identification is discussed.

Bascomb, Shoshana; Manafi, Mammad

1998-01-01

324

Rapid Discrimination of Gram-Positive and Gram-Negative Bacteria in Liquid Samples by Using NaOH-Sodium Dodecyl Sulfate Solution and Flow Cytometry  

PubMed Central

Background For precise diagnosis of urinary tract infections (UTI), and selection of the appropriate prescriptions for their treatment, we explored a simple and rapid method of discriminating gram-positive and gram-negative bacteria in liquid samples. Methodology/Principal Findings We employed the NaOH-sodium dodecyl sulfate (SDS) solution conventionally used for plasmid extraction from Escherichia coli and the automated urine particle analyzer UF-1000i (Sysmex Corporation) for our novel method. The NaOH-SDS solution was used to determine differences in the cell wall structures between gram-positive and gram-negative bacteria, since the tolerance to such chemicals reflects the thickness and structural differences of bacterial cell walls. The UF-1000i instrument was used as a quantitative bacterial counter. We found that gram-negative bacteria, including E. coli, in liquid culture could easily be lysed by direct addition of equal volumes of NaOH-SDS solution. In contrast, Enterococcus faecalis, which is a gram-positive bacterium, could not be completely lysed by the solution. We then optimized the reaction time of the NaOH-SDS treatment at room temperature by using 3 gram-positive and 4 gram-negative bacterial strains and determined that the optimum reaction time was 5 min. Finally, in order to evaluate the generalizability of this method, we treated 8 gram-positive strains and 8 gram-negative strains, or 4 gram-positive and 4 gram-negative strains incubated in voluntary urine from healthy volunteers in the same way and demonstrated that all the gram-positive bacteria were discriminated quantitatively from gram negative bacteria using this method. Conclusions/Significance Using our new method, we could easily discriminate gram-positive and gram-negative bacteria in liquid culture media within 10 min. This simple and rapid method may be useful for determining the treatment course of patients with UTIs, especially for those without a prior history of UTIs. The method may be easily applied in order to obtain additional information for clinical prescriptions from bacteriuria.

Wada, Atsushi; Kono, Mari; Kawauchi, Sawako; Takagi, Yuri; Morikawa, Takashi; Funakoshi, Kunihiro

2012-01-01

325

Aerobic, Selenium-Utilizing Bacillus Isolated from Seeds of Astragalus crotalariae  

PubMed Central

Bacillus sp. strain SS, an aerobic, gram-positive sporeformer, was isolated from seeds of Astragalus crotalariae, a selenium-accumulating plant. This bacillus grew in a nutrient broth (containing beef extract and peptone) if the medium was supplemented with high concentrations of selenium. Concentrations of Na2SeO3 that supported growth ranged from 3 to 100 mM. After 24 h of growth, the culture developed a deep red color characteristic of elemental selenium. When selenium was provided in the form of selenate, the pattern of growth showed a prolonged lag period, from 24 to 48 h. Final growth remained below that of cells cultured in the presence of selenite, and only a light red color developed. Concentrations of selenate below 40 mM failed to support growth. Tellurate, though not tellurite, could replace selenite, but only over a narrow concentration range, 5 to 10 mM. By 24 h, the typical black color of elemental tellurium developed. Bacillus sp. strain SS grew also in brain heart infusion broth and Trypticase soy broth (BBL Microbiology Systems, Cockeysville, Md.) without the addition of selenium or tellurium compounds. When added to these media, 50 mM selenite was tolerated and metabolized by the organism. The crucial distinction between this bacillus and other selenium-tolerant organisms (e.g., Salmonella) remains: under certain conditions, growth requirements of Bacillus sp. strain SS are fulfilled by selenium (and tellurium) compounds. Images

Lindblow-Kull, Carole; Shrift, Alex; Gherna, Robert L.

1982-01-01

326

Antibacterial Activity of Sphingoid Bases and Fatty Acids against Gram-Positive and Gram-Negative Bacteria  

PubMed Central

There is growing evidence that the role of lipids in innate immunity is more important than previously realized. How lipids interact with bacteria to achieve a level of protection, however, is still poorly understood. To begin to address the mechanisms of antibacterial activity, we determined MICs and minimum bactericidal concentrations (MBCs) of lipids common to the skin and oral cavity—the sphingoid bases d-sphingosine, phytosphingosine, and dihydrosphingosine and the fatty acids sapienic acid and lauric acid—against four Gram-negative bacteria and seven Gram-positive bacteria. Exact Kruskal-Wallis tests of these values showed differences among lipid treatments (P < 0.0001) for each bacterial species except Serratia marcescens and Pseudomonas aeruginosa. d-Sphingosine (MBC range, 0.3 to 19.6 ?g/ml), dihydrosphingosine (MBC range, 0.6 to 39.1 ?g/ml), and phytosphingosine (MBC range, 3.3 to 62.5 ?g/ml) were active against all bacteria except S. marcescens and P. aeruginosa (MBC > 500 ?g/ml). Sapienic acid (MBC range, 31.3 to 375.0 ?g/ml) was active against Streptococcus sanguinis, Streptococcus mitis, and Fusobacterium nucleatum but not active against Escherichia coli, Staphylococcus aureus, S. marcescens, P. aeruginosa, Corynebacterium bovis, Corynebacterium striatum, and Corynebacterium jeikeium (MBC > 500 ?g/ml). Lauric acid (MBC range, 6.8 to 375.0 ?g/ml) was active against all bacteria except E. coli, S. marcescens, and P. aeruginosa (MBC > 500 ?g/ml). Complete killing was achieved as early as 0.5 h for some lipids but took as long as 24 h for others. Hence, sphingoid bases and fatty acids have different antibacterial activities and may have potential for prophylactic or therapeutic intervention in infection.

Fischer, Carol L.; Drake, David R.; Dawson, Deborah V.; Blanchette, Derek R.; Brogden, Kim A.

2012-01-01

327

Antibacterial activity of sphingoid bases and fatty acids against Gram-positive and Gram-negative bacteria.  

PubMed

There is growing evidence that the role of lipids in innate immunity is more important than previously realized. How lipids interact with bacteria to achieve a level of protection, however, is still poorly understood. To begin to address the mechanisms of antibacterial activity, we determined MICs and minimum bactericidal concentrations (MBCs) of lipids common to the skin and oral cavity--the sphingoid bases D-sphingosine, phytosphingosine, and dihydrosphingosine and the fatty acids sapienic acid and lauric acid--against four Gram-negative bacteria and seven Gram-positive bacteria. Exact Kruskal-Wallis tests of these values showed differences among lipid treatments (P < 0.0001) for each bacterial species except Serratia marcescens and Pseudomonas aeruginosa. D-sphingosine (MBC range, 0.3 to 19.6 ?g/ml), dihydrosphingosine (MBC range, 0.6 to 39.1 ?g/ml), and phytosphingosine (MBC range, 3.3 to 62.5 ?g/ml) were active against all bacteria except S. marcescens and P. aeruginosa (MBC > 500 ?g/ml). Sapienic acid (MBC range, 31.3 to 375.0 ?g/ml) was active against Streptococcus sanguinis, Streptococcus mitis, and Fusobacterium nucleatum but not active against Escherichia coli, Staphylococcus aureus, S. marcescens, P. aeruginosa, Corynebacterium bovis, Corynebacterium striatum, and Corynebacterium jeikeium (MBC > 500 ?g/ml). Lauric acid (MBC range, 6.8 to 375.0 ?g/ml) was active against all bacteria except E. coli, S. marcescens, and P. aeruginosa (MBC > 500 ?g/ml). Complete killing was achieved as early as 0.5 h for some lipids but took as long as 24 h for others. Hence, sphingoid bases and fatty acids have different antibacterial activities and may have potential for prophylactic or therapeutic intervention in infection. PMID:22155833

Fischer, Carol L; Drake, David R; Dawson, Deborah V; Blanchette, Derek R; Brogden, Kim A; Wertz, Philip W

2012-03-01

328

A Novel p-Nitrophenol Degradation Gene Cluster from a Gram-Positive Bacterium, Rhodococcus opacus SAO101  

PubMed Central

p-Nitrophenol (4-NP) is recognized as an environmental contaminant; it is used primarily for manufacturing medicines and pesticides. To date, several 4-NP-degrading bacteria have been isolated; however, the genetic information remains very limited. In this study, a novel 4-NP degradation gene cluster from a gram-positive bacterium, Rhodococcus opacus SAO101, was identified and characterized. The deduced amino acid sequences of npcB, npcA, and npcC showed identity with phenol 2-hydroxylase component B (reductase, PheA2) of Geobacillus thermoglucosidasius A7 (32%), with 2,4,6-trichlorophenol monooxygenase (TcpA) of Ralstonia eutropha JMP134 (44%), and with hydroxyquinol 1,2-dioxygenase (ORF2) of Arthrobacter sp. strain BA-5-17 (76%), respectively. The npcB, npcA, and npcC genes were cloned into pET-17b to construct the respective expression vectors pETnpcB, pETnpcA, and pETnpcC. Conversion of 4-NP was observed when a mixture of crude cell extracts of Escherichia coli containing pETnpcB and pETnpcA was used in the experiment. The mixture converted 4-NP to hydroxyquinol and also converted 4-nitrocatechol (4-NCA) to hydroxyquinol. Furthermore, the crude cell extract of E. coli containing pETnpcC converted hydroxyquinol to maleylacetate. These results suggested that npcB and npcA encode the two-component 4-NP/4-NCA monooxygenase and that npcC encodes hydroxyquinol 1,2-dioxygenase. The npcA and npcC mutant strains, SDA1 and SDC1, completely lost the ability to grow on 4-NP as the sole carbon source. These results clearly indicated that the cloned npc genes play an essential role in 4-NP mineralization in R. opacus SAO101.

Kitagawa, Wataru; Kimura, Nobutada; Kamagata, Yoichi

2004-01-01

329

Real-Time PCR quantification of PAH-ring hydroxylating dioxygenase (PAH-RHD ?) genes from Gram positive and Gram negative bacteria in soil and sediment samples  

Microsoft Academic Search

Real-Time PCR based assays were developed to quantify Gram positive (GP) and Gram negative (GN) bacterial populations that are capable of degrading the polycyclic aromatic hydrocarbons (PAH) in soil and sediment samples with contrasting contamination levels. These specific and sensitive Real-Time PCR assays were based on the quantification of the copy number of the gene that encodes the alpha subunit

Aurélie Cébron; Marie-Paule Norini; Thierry Beguiristain; Corinne Leyval

2008-01-01

330

Interaction between Penicillin Clindamycin or Metronidazole and Gentamicin Against Species of Clostridia and Anaerobic and Facultatively Anaerobic Gram-Positive cocci.  

National Technical Information Service (NTIS)

Seven anaerobic and facultative Gram-positive cocci and 12 clostridial species were tested for in-vitro and in-vivo susceptibilities to penicillin, clindamycin, and metronidazole, used singly or in combination with gentamicin. The in-vitro tests consisted...

I. Brook R. I. Walker

1985-01-01

331

In vivo efficacy of trovafloxacin (CP-99,219), a new quinolone with extended activities against gram-positive pathogens, Streptococcus pneumoniae, and Bacteroides fragilis.  

PubMed Central

The interesting in vitro antimicrobial activity and pharmacokinetics of the new quinolone trovafloxacin (CP-99,219) warranted further studies to determine its in vivo efficacy in models of infectious disease. The significance of the pharmacokinetic and in vitro antimicrobial profiles of trovafloxacin was shown through efficacy in a series of animal infection models by employing primarily oral therapy. Against acute infections, trovafloxacin was consistently more effective than temafloxacin, ciprofloxacin, and ofloxacin against Streptococcus pneumoniae and other gram-positive pathogens while maintaining activity comparable to that of ciprofloxacin against gram-negative organisms. In a model of murine pneumonia, trovafloxacin was more efficacious than temafloxacin, while ciprofloxacin failed against S. pneumoniae (50% protective doses, 2.1, 29.5, and >100 mg/kg, respectively). In addition to its inherent in vitro potency advantage against S. pneumoniae, these data were supported by a pharmacokinetic study that showed levels of trovafloxacin in pulmonary tissue of S. pneumoniae-infected CF1 mice to be considerably greater than those of temafloxacin and ciprofloxacin (twice the maximum drug concentration in serum; two to three times the half-life, and three to six times the area under the concentration-time curve). Against localized mixed anaerobic infections, trovafloxacin was the only agent to effectively reduce the numbers of recoverable CFU of Bacteroides fragilis ( >1,000-fold), Staphylococcus aureus (1,000-fold), and Escherichia coli ( >100-fold) compared with ciprofloxacin, vancomycin, metronidazole, clindamycin, cefoxitin, and ceftriaxone. The in vitro and in vivo antimicrobial activities of trovafloxacin and its pharmacokinetics in laboratory animals provide support for the ongoing and planned human phase II and III clinical trials.

Girard, A E; Girard, D; Gootz, T D; Faiella, J A; Cimochowski, C R

1995-01-01

332

Identification, Evolution, and Essentiality of the Mevalonate Pathway for Isopentenyl Diphosphate Biosynthesis in Gram-Positive Cocci  

Microsoft Academic Search

The mevalonate pathway and the glyceraldehyde 3-phosphate (GAP)-pyruvate pathway are alternative routes for the biosynthesis of the central isoprenoid precursor, isopentenyl diphosphate. Genomic analysis revealed that the staphylococci, streptococci, and enterococci possess genes predicted to encode all of the enzymes of the mevalonate pathway and not the GAP-pyruvate pathway, unlike Bacillus subtilis and most gram-negative bacteria studied, which possess only

E. IMOGEN WILDING; JAMES R. BROWN; ALEXANDER P. BRYANT; ALISON F. CHALKER; DAVID J. HOLMES; KAREN A. INGRAHAM; SERBAN IORDANESCU; CHI Y. SO; MARTIN ROSENBERG; MICHAEL N. GWYNN

2000-01-01

333

Alginase enzyme production by Bacillus circulans.  

PubMed

Stream and soil samples were screened for microorganisms that would use alginate from mucoid Pseudomonas aeruginosa as the sole carbon and energy source. A pure culture containing large aerobic rods was isolated. The cells were about 0.8 by 2.5 microns in size, had lateral or peritrichous flagella, had a negative Gram stain reaction, and produced spores on sporulation medium. Purified DNA was approximately 46 mol% G+C as measured by thermal denaturation. From these and other biochemical tests, the organism was identified as Bacillus circulans. The enzyme activity that degraded alginate appeared in the culture medium. Upon gel filtration, alginase activity eluted as a single peak at a position corresponding to a protein of 40,000 daltons. Activity recovered from this one-step, partial purification showed apparent endomannuronidase specificity. Like other alginases previously reported, the enzyme appeared likely to be a lyase (or eliminase). However, no Bacillus species or other gram-positive bacteria have heretofore been reported to produce extracellular enzymes with alginase activity. Several other B. circulans strains from the American Type Culture Collection also appeared to have inducible extracellular alginase activity. PMID:6426387

Hansen, J B; Doubet, R S; Ram, J

1984-04-01

334

Alginase enzyme production by Bacillus circulans.  

PubMed Central

Stream and soil samples were screened for microorganisms that would use alginate from mucoid Pseudomonas aeruginosa as the sole carbon and energy source. A pure culture containing large aerobic rods was isolated. The cells were about 0.8 by 2.5 microns in size, had lateral or peritrichous flagella, had a negative Gram stain reaction, and produced spores on sporulation medium. Purified DNA was approximately 46 mol% G+C as measured by thermal denaturation. From these and other biochemical tests, the organism was identified as Bacillus circulans. The enzyme activity that degraded alginate appeared in the culture medium. Upon gel filtration, alginase activity eluted as a single peak at a position corresponding to a protein of 40,000 daltons. Activity recovered from this one-step, partial purification showed apparent endomannuronidase specificity. Like other alginases previously reported, the enzyme appeared likely to be a lyase (or eliminase). However, no Bacillus species or other gram-positive bacteria have heretofore been reported to produce extracellular enzymes with alginase activity. Several other B. circulans strains from the American Type Culture Collection also appeared to have inducible extracellular alginase activity. Images

Hansen, J B; Doubet, R S; Ram, J

1984-01-01

335

Long-term fertilization of organic manure led to the succession of Bacillus community in an alluvial-aquic soil  

NASA Astrophysics Data System (ADS)

Long-term fertilization inevitably influences soil physic-chemical and biological properties. Our previous studies with a long-term fertilization experiment on an alluvial-aquic have revealed that specific Bacillus spp. was observed in organic manure-fertilized soils. The current study investigated the effects of long-term fertilization on the succession of Bacillus community in soils and their functions. The experiment included three fertilizer treatments: organic manure (OM), mineral fertilizers (NPK) and the control (without fertilizers). The results showed that long-term application of chemical fertilizers didn't increase the quantity of soil microbial population as much as organic fertilizers did, but it played an important role in maintaining the diversity and community structure of indigenous Bacilli. Correspondingly, long-term application of organic manure significantly increased the quantity while significantly decreased the diversity of Bacilli community. The ratio of Bacilli/bacteria was more constant in OM treatment than NPK indicating the stability of the response to long-term organic fertilizers. PCR-DGGE and clone library revealed the succession of Bacillus community after long-term application of organic manure and the dominant Bacillus spp occurred in the treatmen OM was Bacillus asahii. Our results also proved that Bacillus asahii was not derived from exogenous organic manure, but one of indigenous bacteria in the soil. Bacillus asahii was induced by the substrate after the application of organic manure, and gradually evolved into dominant Bacillus after 4 to 5 years. With an enzyme assay test of pure species and a soil incubation experiment, we came to a preliminary judgment, that the dominant Bacillus asahii didn't significantly influence the decomposition rate of cellulose and protein in the soil, but it promoted the decomposition of lipids, and could also improve the transformation process from fresh organic matter to humus. Applied organic manure led to the succession of soil microbial community, as a response, the changed microbial community and their activities influenced the turnover of exogenous and native soil organic matter, as well as the residuals of decomposition and microbial metabolisms.

Chen, Ruirui; Lin, Xiangui; Feng, Youzhi; Hu, Junli; Wang, Ruirui

2014-05-01

336

Complete Genome Sequence of Bacillus subtilis Strain PY79  

PubMed Central

Bacillus subtilis is a Gram-positive soil-dwelling and endospore-forming bacterium in the phylum Firmicutes. B. subtilis strain PY79 is a prototrophic laboratory strain that has been highly used for studying a wide variety of cellular pathways. Here, we announce the complete whole-genome sequence of B. subtilis PY79.

Schroeder, Jeremy W.

2013-01-01

337

Pharmacodynamics of TD-1792, a novel glycopeptide-cephalosporin heterodimer antibiotic used against Gram-positive bacteria, in a neutropenic murine thigh model.  

PubMed

TD-1792 is a novel glycopeptide-cephalosporin heterodimer investigational antibiotic that displays potent bactericidal effects against clinically relevant Gram-positive organisms in vitro. The present studies evaluated the in vivo pharmacokinetics (PK) and pharmacodynamics (PD) of TD-1792 in the neutropenic murine thigh infection animal model. TD-1792, dosed subcutaneously (SC), produced dose-dependent reduction in the thigh bacterial burden of several organisms, including methicillin-susceptible and -resistant strains of Staphylococcus aureus and Staphylococcus epidermidis (MSSA, MRSA, MSSE, MRSE, respectively), penicillin-susceptible strains of Streptococcus pneumoniae (PSSP), Streptococcus pyogenes, and vancomycin-intermediate-susceptible Staphylococcus aureus (VISA). In single-dose efficacy studies, the 1-log(10) CFU kill effective dose (ED(1-log kill)) estimates for TD-1792 ranged from 0.049 to 2.55 mg/kg of body weight administered SC, and the bacterial burden was reduced by up to 3 log(10) CFU/g from pretreatment values. Against S. aureus ATCC 33591 (MRSA), the total 24-h log(10) stasis dose (ED(stasis)) and ED(1-logkill) doses for TD-1792 were 0.53 and 1.11 mg/kg/24 h, respectively, compared to 23.4 and 54.6 mg/kg/24 h for vancomycin, indicating that TD-1762 is 44- to 49-fold more potent than vancomycin. PK-PD analysis of data from single-dose and dose-fractionation studies for MRSA (ATCC 33591) demonstrated that the total-drug 24-h area under the concentration-time curve-to-MIC ratio (AUC/MIC ratio) was the best predictor of efficacy (r(2) = 0.826) compared to total-drug maximum plasma concentration of drug-to-MIC ratio (Cmax/MIC ratio; r(2) = 0.715) and percent time that the total-drug plasma drug concentration remains above the MIC (%Time>MIC; r(2) = 0.749). The magnitudes of the total-drug AUC/MIC ratios associated with net bacterial stasis, a 1-log(10) CFU reduction from baseline and near maximal effect, were 21.1, 37.2, and 51.8, respectively. PK-PD targets based on such data represent useful inputs for analyses to support dose selection decisions for clinical studies of patients. PMID:22155835

Hegde, Sharath S; Okusanya, Olanrewaju O; Skinner, Robert; Shaw, Jeng-Pyng; Obedencio, Glenmar; Ambrose, Paul G; Blais, Johanne; Bhavnani, Sujata M

2012-03-01

338

The essential YycFG two-component system of Bacillus subtilis.  

PubMed

The YycFG two-component system, highly conserved in the low G+C gram positives, is essential for cell viability in most organisms in which it has been studied. The system is organized within an operon that includes at least one but often three to four other genes. Products of two of these genes, yycH and yycI, have been shown to have a regulatory role on this two-component system. Immunofluorescent studies identified YycG kinase localization at the cell division sites consistent with its role in regulating cell divisional processes. The essential nature and operon organization of this system commanded special requirements in studying this system genetically. This chapter presents methods utilized in identifying the regulatory circuit that controls the activity of the YycG kinase in Bacillus subtilis. Most aspects of our approaches are applicable to other two-component systems in B. subtilis and the gram positives. Some are limited to essential systems, such as the YycFG system. PMID:17628151

Szurmant, Hendrik; Fukushima, Tatsuya; Hoch, James A

2007-01-01

339

[Evaluation of rapid genotype assay for the identification of gram-positive cocci from blood cultures and detection of mecA and van genes].  

PubMed

Rapid and accurate identification of bacterial pathogens grown in blood cultures of patients with sepsis is crucial for prompt initiation of appropriate therapy in order to decrease related morbidity and mortality rates. Although current automated blood culture systems led to a significant improvement in bacterial detection time, more rapid identification systems are still needed to optimise the establishment of treatment. Novel genotype technology which is developed for the rapid diagnosis of sepsis, is a molecular genetic assay based on DNA multiplex amplification with biotinylated primers followed by hybridization to membrane bound probes. The aim of this study was to evaluate the performance of "Genotype® BC gram-positive? test for the identification of gram-positive cocci grown in blood cultures and rapid detection of mecA and van genes. This test uses DNA.STRIP® technology which includes a panel of probes for identification of 17 gram-positive bacterial species and is able to determinate the methicillin and vancomycin resistance mediating genes (mecA and vanA, vanB, vanC1, vanC2/C3) simultaneously, in a single test run. A total of 55 positive blood cultures from BACTECTM Plus/F (Becton Dickinson, USA) aerobic and pediatric blood culture vials were included in the study. The isolates which exhibit gram-positive coccus morphology by Gram staining were identified by Genotype ® BC gram-positive test (Hain Life Science, Germany). All of the samples were also identified with the use of Phoenix PMIC/ID Panel (Becton Dickinson, USA) and antibiotic susceptibilities were determined. Of the 55 blood culture isolates, 17 were identified as Staphylococcus epidermidis [all were methicillin-resistant (MR)], 9 were S.aureus (one was MR), 18 were S.hominis (10 were MR), 4 were E.faecalis, 3 were E. faecium (one was vanconycin-resistant), 2 were S.saprophyticus (one was MR), 1 was S.warneri and 1 was S.haemolyticus, by Phoenix automated system. Genotype® BC gram-positive test results revealed consistency with Phoenix system regarding bacterial identification in 46 (83.6%) of the samples. The two bacteria identified as S.saprophyticus by the Phoenix system could not be identified by the Genotype® BC test since this species were not included in the identification panel of the system, however, mecA gene were detected in these two samples by Genotype® BC test. Genotype® BC test detected mecA gene in five samples which were not detected as methicillin resistant by the Phoenix system. Besides polymicrobial growth was determined in five samples by Genotype ® BC test, but not by the automated system. One E.faecium isolate with vanA gene was correctly identified by Genotype® BC test. In conclusion, Genotype® BC gram-positive test is a fast and reliable test for the identification of the most important gram-positive pathogens and mecA and van genes directly from positive blood culture bottles. This test was also found superior than the automated Phoenix system regarding the detection of polymicrobial growth. These data indicated that, routine use of DNA strip technology-based assay would be useful for clinical diagnosis in patients with sepsis. PMID:22090289

Gülhan, Bar??; Atmaca, Selahattin; Ozekinci, Tuncer; Suay, Adnan

2011-10-01

340

Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp. RN2  

PubMed Central

The aim of this study was to characterize the organic solvent and detergent tolerant properties of recombinant lipase isolated from thermotolerant Bacillus sp. RN2 (Lip-SBRN2). The isolation of the lipase-coding gene was achieved by the use of inverse and direct PCR. The complete DNA sequencing of the gene revealed that the lip-SBRN2 gene contains 576 nucleotides which corresponded to 192 deduced amino acids. The purified enzyme was homogeneous with the estimated molecular mass of 19 kDa as determined by SDS-PAGE and gel filtration. The Lip-SBRN2 was stable in a pH range of 9–11 and temperature range of 45–60 °C. The enzyme was a non metallo-monomeric protein and was active against pNP-caprylate (C8) and pNP-laurate (C12) and coconut oil. The Lip-SBRN2 exhibited a high level of activity in the presence of 108% benzene, 102.4% diethylether and 112% SDS. It is anticipated that the organic solvent and detergent tolerant enzyme secreted by Bacillus sp. RN2 will be applicable as catalysts for reaction in the presence of organic solvents and detergents.

Kanjanavas, Pornpimon; Khuchareontaworn, Sintawee; Khawsak, Paisarn; Pakpitcharoen, Arda; Pothivejkul, Khajeenart; Santiwatanakul, Somchai; Matsui, Kenji; Kajiwara, Tadahiko; Chansiri, Kosum

2010-01-01

341

Comparison of Three Commercial Rapid Identification Systems for the Unusual Gram-Positive Cocci Dolosigranulum pigrum, Ignavigranum ruoffiae, and Facklamia Species  

Microsoft Academic Search

We evaluated three rapid identification systems—The Biomerieux rapid ID 32 STREP (ID32), the BBL Crystal rapid gram-positive identification (Crystal), and the Remel IDS RapID STR (IDS) systems—for their ability to identify 7 strains of Alloiococcus otitidis, 27 strains of Dolosigranulum pigrum, 3 strains of Ignavigranum ruoffiae, and 18 strains of 4 different Facklamia species. Since none of these six species

Leslye L. LaClaire; Richard R. Facklam; Brian D. Plikaytis; David Goldblatt; Carl E. Frasch; Christine Blondeau; Michael J. Bybel; G. Scott Giebink; Ingileif Jonsdottir; Helena Käyhty; Helle Bossen Konradsen; Dace V. Madore; Moon H. Nahm; Cheryl A. Schulman; Patricia F. Holder; Tamar Lezhava; Cheryl M. Elie; George M. Carlone

2000-01-01

342

In Vitro Activities of Daptomycin, Vancomycin, Quinupristin- Dalfopristin, Linezolid, and Five Other Antimicrobials against 307 Gram-Positive Anaerobic and 31 Corynebacterium Clinical Isolates  

Microsoft Academic Search

The activities of daptomycin, a cyclic lipopeptide, and eight other agents were determined against 338 strains of gram-positive anaerobic bacteria and corynebacteria by the NCCLS reference agar dilution method with supplemented brucella agar for the anaerobes and Mueller-Hinton agar for the corynebacteria. The dapto- mycin MICs determined on Ca2-supplemented (50 mg\\/liter) brucella agar plates were one- to fourfold lower than

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

2003-01-01

343

Detection of heavy metal ion resistance genes in Gram-positive and Gram-negative bacteria isolated from a lead-contaminated site  

Microsoft Academic Search

Resistance to a range of heavy metal ions wasdetermined for lead-resistant and other bacteria whichhad been isolated from a battery-manufacturing sitecontaminated with high concentrations of lead. Several Gram-positive (belonging to the genera Arthrobacter and Corynebacterium) andGram-negative (Alcaligenes species) isolateswere resistant to lead, mercury, cadmium, cobalt,zinc and copper, although the levels of resistance tothe different metal ions were specific for eachisolate.

Suzana Trajanovska; Margaret L. Britz; Mrinal Bhave

1997-01-01

344

In vitro activity of XF-73, a novel antibacterial agent, against antibiotic-sensitive and -resistant Gram-positive and Gram-negative bacterial species  

Microsoft Academic Search

The antibacterial activity of XF-73, a dicationic porphyrin drug, was investigated against a range of Gram-positive and Gram-negative bacteria with known antibiotic resistance profiles, including resistance to cell wall synthesis, protein synthesis, and DNA and RNA synthesis inhibitors as well as cell membrane-active antibiotics. Antibiotic-sensitive strains for each of the bacterial species tested were also included for comparison purposes. XF-73

David J. Farrell; Marion Robbins; William Rhys-Williams; William G. Love

2010-01-01

345

Lactobacillus plantarum gene clusters encoding putative cell-surface protein complexes for carbohydrate utilization are conserved in specific gram-positive bacteria  

Microsoft Academic Search

BACKGROUND: Genomes of gram-positive bacteria encode many putative cell-surface proteins, of which the majority has no known function. From the rapidly increasing number of available genome sequences it has become apparent that many cell-surface proteins are conserved, and frequently encoded in gene clusters or operons, suggesting common functions, and interactions of multiple components. RESULTS: A novel gene cluster encoding exclusively

Roland Siezen; Jos Boekhorst; Lidia Muscariello; Douwe Molenaar; Bernadet Renckens; Michiel Kleerebezem

2006-01-01

346

In vitro Activity of Monoclonal and Recombinant Yeast Killer Toxin-like Antibodies Against Antibiotic-resistant Gram-positive Cocci  

Microsoft Academic Search

Background: Monoclonal (mAbKT) and recom- binant single-chain (scFvKT) anti-idiotypic anti- bodies were produced to represent the internal image of a yeast killer toxin (KT) characterized by a wide spectrum of antimicrobial activity, including Gram-positive cocci. Pathogenic eukaryotic and prokaryotic microorganisms, such as Candida albi- cans, Pneumocystis carinii, and a multidrug-resistant strain of Mycobacterium tuberculosis, presenting spe- cific, although yet undefined,

S. Conti; W. Magliani; S. Arseni; E. Dieci; A. Salati; P. E. Varaldo; L. Polonelli

2000-01-01

347

Kineococcus aurantiacus gen. nov., sp. nov., a New Aerobic, Gram-Positive, Motile Coccus with rneso-Diaminopimelic Acid and Arabinogalactan in the Cell Wall  

Microsoft Academic Search

A new aerobic, gram-positive, motile coccus isolated from soil is described. Strain RA 333= (T = type strain) has the following characteristics: menaquinone MK-9(H2); G+C content of DNA of 73.9 mol%; meso- diaminopimelic acid, glutamic acid, and alanine in a molar ratio of ca. 1:1:2 (type Aly); and arabinose and galactose in the cell wall. The two sugars are contained

AKIRA YOKOTA; TOMOHIKO TAMURA; TADASHI NISHII; TORU HASEGAWA

348

Comparison between the photocatalytic inactivation of Gram-positive E. faecalis and Gram-negative E. coli faecal contamination indicator microorganisms  

Microsoft Academic Search

Photocatalytic inactivation of two different faecal contamination indicator microorganisms, the Gram-negative Escherichia coli and the Gram-positive Enterococcus faecalis, has been studied using TiO2 in suspension and immobilized onto the reactor wall. The effect of the main variables of the photocatalytic process on the disinfection efficiency has been analyzed using deionized water and a simulated effluent of wastewater treatment plant (WTP).

Rafael van Grieken; Javier Marugán; Cristina Pablos; Laura Furones; Ainhoa López

2010-01-01

349

Photoinactivation of bacteria. Use of a cationic water-soluble zinc phthalocyanine to photoinactivate both Gram-negative and Gram-positive bacteria  

Microsoft Academic Search

The photosensitization of microorganisms is potentially useful for sterilization and for the treatment of certain bacterial diseases. Unit now, any broad spectrum approach has been inhibited because, although Gram-positive bacteria can be photoinactivated by a range of photosensitizer, Gram-negative bacteria have not usually been susceptible to photosensitized destruction.In the present work, it has been shown that the Gram-negative bacteria Escheria

Andrew Minnock; David I. Vernon; Jack Schofield; John Griffiths; J. Howard Parish; Stanley B. Brown

1996-01-01

350

Obtaining and characterization of DNA-containing micromummies of yeasts and gram-positive bacteria with enhanced cell wall permeability: Application in PCR  

Microsoft Academic Search

The procedure of obtaining DNA-containing cell envelopes (“micromummies”) of bacteria, yeasts, and fungi using chaotropic\\u000a salts has been developed previously and the possibility of their direct application in PCR has been demonstrated. The fine\\u000a structure of micromummies has been studied by electron microscopic methods. This work has demonstrated that additional treatment\\u000a of micromummies of yeasts and gram-positive bacteria with proteinase

V. N. Danilevich; V. I. Duda; N. E. Suzina; E. V. Grishin

2007-01-01

351

Activity of daptomycin against susceptible and multidrug-resistant Gram-positive pathogens collected in the SECURE study (Europe) during 2000-2001  

Microsoft Academic Search

Antibiotic resistance was prevalent in Gram-positive pathogens collected from 40 sites in 15 European countries during 2000-2001. Among Staphylococcus aureus, 27.3% of all isolates submitted were resistant to oxacillin and ranged from 0% of isolates from the Netherlands to 36.9% of isolates from Portugal. The overall prevalence of vancomycin-resistant Enterococcus faecium was 25.1%, with Italy submitting the largest percentage of

Ian A. Critchley; Deborah C. Draghi; Daniel F. Sahm; Clyde Thornsberry; Mark E. Jones; James A. Karlowsky

352

Differential Roles of TLR2 and TLR4 in Recognition of Gram-Negative and Gram-Positive Bacterial Cell Wall Components  

Microsoft Academic Search

Toll-like receptor (TLR) 2 and TLR4 are implicated in the recognition of various bacterial cell wall components, such as lipopolysaccharide (LPS). To investigate in vivo roles of TLR2, we generated TLR2-deficient mice. In contrast to LPS unresponsiveness in TLR4-deficient mice, TLR2-deficient mice responded to LPS to the same extent as wild-type mice. TLR2-deficient macrophages were hyporesponsive to several Gram-positive bacterial

Osamu Takeuchi; Katsuaki Hoshino; Taro Kawai; Hideki Sanjo; Haruhiko Takada; Tomohiko Ogawa; Kiyoshi Takeda; Shizuo Akira

1999-01-01

353

Synthesis of well-dispersed silver nanorods of different aspect ratios and their antimicrobial properties against gram positive and negative bacterial strains  

PubMed Central

In the present contribution, we describe the synthesis of highly dispersed silver nanorods (NRs) of different aspect ratios using a chemical route. The shape and size of the synthesized NRs were characterized by Transmission Electron Microscopy (TEM) and UV-visible spectroscopy. Longitudinal and transverse absorptions bands confirm the rod type structure. The experimentally recorded UV-visible spectra of NRs solutions were fitted by using an expression of the extinction coefficient for rod like nano structures under the dipole approximation. Simulated and experimentally observed UV-visible spectra were compared to determine the aspect ratios (R) of NRs. The average values of R for NR1, NR2 and NR3 solutions are estimated to be 3.0?±?0.1, 1.8?±?0.1 and 1.2?±?0.1, respectively. These values are in good agreement with those obtained by TEM micrographs. The silver NRs of known aspect ratios are used to study antimicrobial activities against B. subtilis (gram positive) and E. coli (gram negative) microbes. We observed that the NRs of intermediate aspect ratio (R?=?1.8) have greater antimicrobial effect against both, B. subtilis (gram positive) and E. coli (gram negative). The NRs of aspect ratio, R?=?3.0 has better antimicrobial activities against gram positive than on the gram negative.

2013-01-01

354

A Disulfide Bond-Containing Alkaline Phosphatase Triggers a BdbC-Dependent Secretion Stress Response in Bacillus subtilis  

Microsoft Academic Search

The gram-positive bacterium Bacillus subtilis secretes high levels of proteins into its environment. Most of these secretory proteins are exported from the cytoplasm in an unfolded state and have to fold efficiently after membrane translocation. As previously shown for ?-amylases of Bacillus species, inefficient posttranslocational protein folding is potentially detrimental and stressful. In B. subtilis, this so-called secretion stress is

Elise Darmon; Ronald Dorenbos; Jochen Meens; Roland Freudl; Haike Antelmann; Michael Hecker; Oscar P. Kuipers; Sierd Bron; Wim J. Quax; Jean-Yves F. Dubois; Jan Maarten van Dijl

2006-01-01

355

Display of proteins on Bacillus subtilis endospores  

Microsoft Academic Search

The targeting and anchoring of heterologous proteins and peptides to the outer surface of bacteriophages and cells is becoming\\u000a increasingly important, and has been employed as a tool for fundamental and applied research in microbiology, molecular biology,\\u000a vaccinology, and biotechnology. Less known are endospores or spores produced by some Gram-positive species. Spores of Bacillus subtilis are surrounded by a spore

Junehyung Kim; Wolfgang Schumann

2009-01-01

356

Microbe forensics: Oxygen and hydrogen stable isotope ratios in Bacillus subtilis cells and spores  

PubMed Central

Bacillus subtilis, a Gram-positive, endospore-forming soil bacterium, was grown in media made with water of varying oxygen (?18O) and hydrogen (?D) stable isotope ratios. Logarithmically growing cells and spores were each harvested from the cultures and their ?18O and ?D values determined. Oxygen and hydrogen stable isotope ratios of organic matter were linearly related with those of the media water. We used the relationships determined in these experiments to calculate the effective whole-cell fractionation factors between water and organic matter for B. subtilis. We then predicted the ?18O and ?D values of spores produced in nutritionally identical media and local water sources for five different locations around the United States. Each of the measured ?18O and ?D values of the spores matched the predicted values within a 95% confidence interval, indicating that stable isotope ratio analyses may be a powerful tool for tracing the geographic point-of-origin for microbial products.

Kreuzer-Martin, Helen W.; Lott, Michael J.; Dorigan, Janet; Ehleringer, James R.

2003-01-01

357

Molecular study of a squalene cyclase homolog gene in Bacillus subtilis  

NASA Astrophysics Data System (ADS)

Polycyclic triterpenoids such as hopanes and steranes are formed by enzymatic cyclization of linear isoprenoid precursors by squalene cyclases and oxidosqualene cyclases. Due to their amazing preservation potential, polycyclic triterpenoids have been used to indicate the source of organic matter in oils and sediments for decades, although many cannot be attributed to known organisms and genes. To bridge the gap between the genomic database and the geochemical record, we are using molecular tools to study the expression, intracellular localization, and products of a squalene cyclase homolog found in Bacillus subtilis, a Gram-positive soil bacterium. We find that the gene is expressed during sporulation and is localized to the spore coat. Our results may help to understand the source of some previously unassigned natural products, and they may also provide clues to the physiological role of triterpenoids in the Bacillales.

Bosak, T.; Pearson, A.; Losick, R.

2005-12-01

358

Protein secretion in Bacillus species.  

PubMed Central

Bacilli secrete numerous proteins into the environment. Many of the secretory proteins, their export signals, and their processing steps during secretion have been characterized in detail. In contrast, the molecular mechanisms of protein secretion have been relatively poorly characterized. However, several components of the protein secretion machinery have been identified and cloned recently, which is likely to lead to rapid expansion of the knowledge of the protein secretion mechanism in Bacillus species. Comparison of the presently known export components of Bacillus species with those of Escherichia coli suggests that the mechanism of protein translocation across the cytoplasmic membrane is conserved among gram-negative and gram-positive bacteria differences are found in steps preceding and following the translocation process. Many of the secretory proteins of bacilli are produced industrially, but several problems have been encountered in the production of Bacillus heterologous secretory proteins. In the final section we discuss these problems and point out some possibilities to overcome them.

Simonen, M; Palva, I

1993-01-01

359

Comparison of Three Commercial Rapid Identification Systems for the Unusual Gram-Positive Cocci Dolosigranulum pigrum, Ignavigranum ruoffiae, and Facklamia Species  

PubMed Central

We evaluated three rapid identification systems—The Biomerieux rapid ID 32 STREP (ID32), the BBL Crystal rapid gram-positive identification (Crystal), and the Remel IDS RapID STR (IDS) systems—for their ability to identify 7 strains of Alloiococcus otitidis, 27 strains of Dolosigranulum pigrum, 3 strains of Ignavigranum ruoffiae, and 18 strains of 4 different Facklamia species. Since none of these six species of gram-positive cocci are included in the identification databases for these systems, the correct identification for the strains tested should be “unacceptable ID” for the ID32 and Crystal systems or “no choice” for the IDS system. The ID32 system identified all 27 strains of D. pigrum, 6 of 18 Facklamia species, and 2 of 3 cultures of I. ruoffiae as “unacceptable ID.” The Crystal system identified 10 of 27 D. pigrum, 2 of 18 Facklamia species, and 2 of 3 I. ruoffiae strains as “unacceptable ID.” The IDS system identified only 1 culture of D. pigrum as “no choice,” but it also identified 2 cultures of D. pigrum as a “questionable microcode” and 19 cultures of D. pigrum as an “inadequate ID, E. faecalis 90%, S. intermedius 9%.” A total of 2 of the 18 cultures of Facklamia and all 3 of the I. ruoffiae cultures were correctly identified as “no choice.” The most common misidentifications of Facklamia species by the ID32 and IDS systems were as various Streptococcus species and as Gemella species. In the Crystal system, the most common erroneous identification was Micrococcus luteus. These data indicate the need for the commercial manufacturers of these products to update their databases to include newly described species of gram-positive cocci.

LaClaire, Leslye L.; Facklam, Richard R.

2000-01-01

360

Systematic review of membrane components of gram-positive bacteria responsible as pyrogens for inducing human monocyte/macrophage cytokine release.  

PubMed

Fifty years after the elucidation of lipopolysaccharides (LPS, endotoxin) as the principal structure of Gram-negative bacteria activating the human immune system, its Gram-positive counterpart is still under debate. Pyrogen tests based on the human monocyte activation have been validated for LPS detection as an alternative to the rabbit test and, increasingly, the limulus amebocyte lysate test. For full replacement, international validations with non-endotoxin pyrogens are in preparation. Following evidence-based medicine approaches, a systematic review of existing evidence as to the structural nature of the Gram-positive pyrogen was undertaken. For the three major constituents suggested, i.e., peptidoglycan, lipoteichoic acids (LTA), and bacterial lipoproteins (LP), the questions to be answered and a search strategy for relevant literature was developed, starting in MedLine. The evaluation was based on the Koch-Dale criteria for a mediator of an effect. A total of 380 articles for peptidoglycan, 391 for LP, and 285 for LTA were retrieved of which 12, 8, and 24, respectively, fulfilled inclusion criteria. The compiled data suggest that for peptidoglycan two Koch-Dale criteria are fulfilled, four for LTA, and two for bacterial LP. In conclusion, based on the best currently available evidence, LTA is the only substance that fulfills all criteria. LTA has been isolated from a large number of bacteria, results in cytokine release patterns inducible also with synthetic LTA. Reduction in bacterial cytokine induction with an inhibitor for LTA was shown. However, this systematic review cannot exclude the possibility that other stimulatory compounds complement or substitute for LTA in being the counterpart to LPS in some Gram-positive bacteria. PMID:22529809

Rockel, Christoph; Hartung, Thomas

2012-01-01

361

Comparison of three commercial rapid identification systems for the unusual gram-positive cocci Dolosigranulum pigrum, Ignavigranum ruoffiae, and Facklamia species.  

PubMed

We evaluated three rapid identification systems-The Biomerieux rapid ID 32 STREP (ID32), the BBL Crystal rapid gram-positive identification (Crystal), and the Remel IDS RapID STR (IDS) systems-for their ability to identify 7 strains of Alloiococcus otitidis, 27 strains of Dolosigranulum pigrum, 3 strains of Ignavigranum ruoffiae, and 18 strains of 4 different Facklamia species. Since none of these six species of gram-positive cocci are included in the identification databases for these systems, the correct identification for the strains tested should be "unacceptable ID" for the ID32 and Crystal systems or "no choice" for the IDS system. The ID32 system identified all 27 strains of D. pigrum, 6 of 18 Facklamia species, and 2 of 3 cultures of I. ruoffiae as "unacceptable ID." The Crystal system identified 10 of 27 D. pigrum, 2 of 18 Facklamia species, and 2 of 3 I. ruoffiae strains as "unacceptable ID." The IDS system identified only 1 culture of D. pigrum as "no choice," but it also identified 2 cultures of D. pigrum as a "questionable microcode" and 19 cultures of D. pigrum as an "inadequate ID, E. faecalis 90%, S. intermedius 9%." A total of 2 of the 18 cultures of Facklamia and all 3 of the I. ruoffiae cultures were correctly identified as "no choice." The most common misidentifications of Facklamia species by the ID32 and IDS systems were as various Streptococcus species and as Gemella species. In the Crystal system, the most common erroneous identification was Micrococcus luteus. These data indicate the need for the commercial manufacturers of these products to update their databases to include newly described species of gram-positive cocci. PMID:10834950

LaClaire, L L; Facklam, R R

2000-06-01

362

Rapid identification of Gram-positive pathogens and their resistance genes from positive blood culture broth using a multiplex tandem RT-PCR assay.  

PubMed

The early initiation of targeted antibiotic therapy in patients with bacteraemia and septic shock impacts favourably on outcomes. Rapid methods are therefore increasingly employed for bacterial identification directly from positive blood culture bottles, but with variable success. We evaluated the performance of the Gram Positive 12 multiplex tandem PCR (MT-PCR) assay (AusDiagnostics; catalogue no. 6202, version 07) containing targets for the identification of staphylococci including Staphylococcus aureus, streptococci including Streptococcus pneumoniae, enterococci including Enterococcus faecalis and Enterococcus faecium and their common antibiotic resistance genes (mecA, vanA, vanB). A total of 673 aerobic and anaerobic blood culture broths demonstrating Gram-positive cocci on microscopy were analysed in parallel with traditional phenotypic methods. Amplification of the internal control was inhibited in 79/673 (11.7?%) samples; however, MT-PCR identification was in concordance with phenotypic identification to the genus level in 96.6?% (537/556) of the remaining monomicrobial specimens and to the species level, where applicable, in 100?% (172/172) of samples. MT-PCR identification for 94.7?% (36/38) of polymicrobial samples matched traditional phenotypic identification. Meticillin and vancomycin susceptibility results determined by MT-PCR in blood culture broths demonstrated complete agreement with those determined by phenotypic methods in all 143 Staphylococcus aureus isolates and eight E. faecium isolates, respectively. Gram-positive pathogens and their key antibiotic resistance markers were reliably identified with the MT-PCR assay within 3 h of a positive blood culture result. PMID:23139396

Hazelton, Briony J; Thomas, Lee C; Unver, Tuba; Iredell, Jonathan R

2013-02-01

363

Impact of results of a rapid Staphylococcus aureus diagnostic test on prescribing of antibiotics for patients with clustered gram-positive cocci in blood cultures.  

PubMed

In tropical northern Australia, approximately 20% of Staphylococcus aureus bacteremia is caused by methicillin-resistant Staphylococcus aureus (MRSA). We prospectively evaluated the impact on clinician antibiotic prescribing of the results obtained from performing the GeneXpert MRSA/SA test on 151 positive blood cultures with clustered gram-positive cocci. The GeneXpert result led to earlier appropriate prescription of vancomycin for 54% of patients with MRSA; 25% of patients avoided vancomycin, and 16% of patients had all antibiotics ceased. PMID:22493335

Davies, Jane; Gordon, Claire L; Tong, Steven Y C; Baird, Robert W; Davis, Joshua S

2012-06-01

364

Physico-Chemical-Managed Killing of Penicillin-Resistant Static and Growing Gram-Positive and Gram-Negative Vegetative Bacteria  

NASA Technical Reports Server (NTRS)

Systems and methods for the use of compounds from the Hofmeister series coupled with specific pH and temperature to provide rapid physico-chemical-managed killing of penicillin-resistant static and growing Gram-positive and Gram-negative vegetative bacteria. The systems and methods represent the more general physico-chemical enhancement of susceptibility for a wide range of pathological macromolecular targets to clinical management by establishing the reactivity of those targets to topically applied drugs or anti-toxins.

Richmond, Robert Chaffee (Inventor); Schramm, Jr., Harry F. (Inventor); Defalco, Francis G. (Inventor); Farris, III, Alex F. (Inventor)

2012-01-01

365

Propeptide of Bacillus subtilis amylase enhances extracellular production of human interferon-? in Bacillus subtilis  

Microsoft Academic Search

The Gram-positive bacterium, Bacillus subtilis and related species are widely used industrially as hosts for producing enzymes. These species possess a high potential to\\u000a produce secreted proteins into the culture medium. Nevertheless, the secretion of heterologous proteins by these species is\\u000a frequently inefficient. In this study, the human interferon-?2b (hIFN-?2b) was used as a heterologous model protein, to investigate\\u000a the

Hiroshi Kakeshita; Yasushi Kageyama; Katsutoshi Ara; Katsuya Ozaki; Kouji Nakamura

2011-01-01

366

The RepA_N replicons of Gram-positive bacteria: a family of broadly distributed but narrow host range plasmids  

PubMed Central

The pheromone-responsive conjugative plasmids of Enterococcus faecalis and the multi-resistance plasmids pSK1 and pSK41 of Staphylococcus aureus are among the best studied plasmids native to Gram-positive bacteria. Although these plasmids seem largely restricted to their native hosts, protein sequence comparison of their replication initiator proteins indicates that they are clearly related. Homology searches indicate that these replicons are representatives of a large family of plasmids and a few phage that are widespread among the low G+C Gram-positive bacteria. We propose to name this family the RepA_N family of replicons after the annotated conserved domain that the initiator protein contains. Detailed sequence comparisons indicate that the initiator protein phylogeny is largely congruent with that of the host, suggesting that the replicons have evolved along with their current hosts and that intergeneric transfer has been rare. However, related proteins were identified on chromosomal regions bearing characteristics indicative of ICE elements, and the phylogeny of these proteins displayed evidence of more frequent intergeneric transfer. Comparison of stability determinants associated with the RepA_N replicons suggests that they have a modular evolution as has been observed in other plasmid families.

Weaver, Keith E.; Kwong, Stephen M.; Firth, Neville; Francia, Maria Victoria

2009-01-01

367

The bacterial cytoskeleton and its putative role in membrane vesicle formation observed in a Gram-positive bacterium producing starch-degrading enzymes.  

PubMed

Bacteria may possess various kinds of cytoskeleton. In general, bacterial cytoskeletons may play a role in the control and preservation of the cell shape. Such functions become especially evident when the bacteria do not possess a true wall and are nevertheless elongated (e.g. Mycoplasma spp.) or under extreme cultivation conditions whereby loss of the entire bacterial cell wall takes place. Bacterial cytoskeletons may control and preserve the cell shape only if a number of preconditions are fulfilled. They should be present not only transiently, but permanently, they should be located as a lining close to the inner face of the cytoplasmic membrane, enclosing the entire cytoplasm, and they should comprise structural elements (fibrils) crossing the inner volume of the cell in order to provide the necessary stability for the lining. Complete loss of the cell wall layers had earlier been observed to occur during extensive production of bacterial starch-degrading enzymes in an optimized fermentation process by a Gram-positive bacterium. Even under these conditions, the cells had maintained their elongated shape and full viability. Which of the various kinds of bacterial cytoskeleton might have been responsible for shape preservation? Only one of them, the primary or basic cytoskeleton turns out to fulfil the necessary preconditions listed above. Its structural features now provided a first insight into a possible mechanism of formation of membrane blebs and vesicles as observed in the Gram-positive eubacterium Thermoanaerobacterium thermosulfurogenes EM1, and the putative role of the cytoskeletal web in this process. PMID:15153765

Mayer, Frank; Gottschalk, Gerhard

2003-01-01

368

In vivo metabolism of 2,2 prime -diaminopimelic acid from gram-positive and gram-negative bacterial cells by ruminal microorganisms and ruminants and its use as a marker of bacterial biomass  

SciTech Connect

Cells of Bacillus megaterium GW1 and Escherichia coli W7-M5 were specifically radiolabeled with 2,2{prime}-diamino (G-{sup 3}H) pimelic acid (({sup 3}H)DAP) as models of gram-positive and gram-negative bacteria, respectively. Two experiments were conducted to study the in vivo metabolism of 2,2{prime}-diaminopimelic acid (DAP) in sheep. In experiment 1, cells of ({sup 3}H)DAP-labeled B. megaterium GW1 were infused into the rumen of one sheep and the radiolabel was traced within microbial samples, digesta, and the whole animal. Bacterially bound ({sup 3}H)DAP was extensively metabolized, primarily (up to 70% after 8 h) via decarboxylation to ({sup 3}H)lysine by both ruminal protozoa and ruminal bacteria. Recovery of infused radiolabel in urine and feces was low (42% after 96 h) and perhaps indicative of further metabolism by the host animal. In experiment 2, ({sup 3}H)DAP-labeled B. megaterium GW1 was infused into the rumens of three sheep and ({sup 3}H)DAP-labeled E. coli W7-W5 was infused into the rumen of another sheep. The radioactivity contents of these mutant bacteria were insufficient to use as tracers, but the metabolism of DAP was monitored in the total, free, and peptidyl forms. Free DAP, as a proportion of total DPA in duodenal digesta, varied from 0 to 9.5%, whereas peptidyl DAP accounted for 8.3 to 99.2%.

Masson, H.A.; Denholm, A.M.; Ling, J.R. (Univ. College of Wales (United Kingdom))

1991-06-01

369

Assessing the interactions of a natural antibacterial clay with model Gram-positive and Gram-negative human pathogens  

NASA Astrophysics Data System (ADS)

The emergence of antibiotic resistant bacteria and increasing accumulations of antibiotics in reclaimed water, drive the quest for new natural antimicrobials. We are studying the antibacterial mechanism(s) of clays that have shown an ability to destroy bacteria or significantly inhibit their growth. One possible mode of action is from soluble transition metal species, particularly reduced Fe, capable of generating deleterious oxygen radical species. Yet another possibility is related to membrane damage as a consequence of physical or electrostatic interaction between clay and bacteria. Both mechanisms could combine to produce cell death. This study addresses a natural antibacterial clay from the NW Amazon basin, South America (AMZ clay). Clay mineralogy is composed of disordered kaolinite (28.9%), halloysite (17.8%) illite (12%) and smectite (16.7%). Mean particle size is 1.6?m and total and specific surface area 278.82 and 51.23 m2/g respectively. The pH of a suspension (200mg/ml) is 4.1 and its Eh is 361mV after 24h of equilibration. The ionic strength of the water in equilibrium with the clay after 24 h. is 6 x10-4M. These conditions, affect the element solubility, speciation, and interactions between clay and bacteria. Standard microbiological methods were used to assess the viability of two model bacteria (Escherichia coli and Bacillus subtilis) after incubation with clay at 37 degC for 24 hrs. A threefold reduction in bacterial viability was observed upon treatment with AMZ clay. We separated the cells from the clay using Nycodenz gradient media and observed the mounts under the TEM and SEM. Results showed several membrane anomalies and structural changes that were not observed in the control cells. Additionally, clay minerals appeared in some places attached to cell walls. Experiments showed that exchanging AMZ clay with KCl caused loss of antibacterial property. Among the exchangeable -and potentially toxic- ions we measured Al+3, Cu+2, Zn+2, Ba+2 and Co+2. Besides being toxic at high concentrations, these species affect the electrophoretic interactions between clay and bacteria surfaces. Additionally, the cation exchange neutralizes the clay surface charge thus modifying further the behavior of particles in suspension. Therefore, we evaluated the clay and bacteria zeta potential (?) as an index for possible electrostatic forces and modeled the total interactions using DLVO theory. We suspended the particles in water equilibrated with clay (leachate). Results show that at pH 4, the ? of clays is -14 mV while it is -3mV for bacteria. The divalent ions and trivalent Aluminum, present in the AMZ leachate, compress the thickness of the double layer (hydration shell) thus decreasing electrostatic repulsion and allowing particles to come closer. The proximity of particles increases the probability of attractive forces to bind clays and cells. In summary, results indicate that a process other than simple chemical transfer from clay to bacteria is operating. The electrostatic attraction and physical proximity may enhance the toxic action of metals and interfere with the membrane properties or processes.

Londono, S. C.; Williams, L. B.

2013-12-01

370

Bacteriophages that infect Bacillus bacteria (Anthrax)  

US Patent & Trademark Office Database

The invention provides bacteriophages that infect Bacillus bacteria, including Bacillus anthracis, and compositions containing the bacteriophages. The invention also provides methods for using the bacteriophages of the invention to prevent and treat infection of an organism by Bacillus bacteria. Methods and materials to decontaminate a surface or an organism that is contaminated with Bacillus bacteria or Bacillus spores is also provided.

2008-05-20

371

The Arthromitus stage of Bacillus cereus: intestinal symbionts of animals.  

PubMed

In the guts of more than 25 species of arthropods we observed filaments containing refractile inclusions previously discovered and named "Arthromitus" in 1849 by Joseph Leidy [Leidy, J. (1849) Proc. Acad. Nat. Sci. Philadelphia 4, 225-233]. We cultivated these microbes from boiled intestines of 10 different species of surface-cleaned soil insects and isopod crustaceans. Literature review and these observations lead us to conclude that Arthromitus are spore-forming, variably motile, cultivable bacilli. As long rod-shaped bacteria, they lose their flagella, attach by fibers or fuzz to the intestinal epithelium, grow filamentously, and sporulate from their distal ends. When these organisms are incubated in culture, their life history stages are accelerated by light and inhibited by anoxia. Characterization of new Arthromitus isolates from digestive tracts of common sow bugs (Porcellio scaber), roaches (Gromphodorhina portentosa, Blaberus giganteus) and termites (Cryptotermes brevis, Kalotermes flavicollis) identifies these flagellated, spore-forming symbionts as a Bacillus sp. Complete sequencing of the 16S rRNA gene from four isolates (two sow bug, one hissing roach, one death's head roach) confirms these as the low-G+C Gram-positive eubacterium Bacillus cereus. We suggest that B. cereus and its close relatives, easily isolated from soil and grown on nutrient agar, enjoy filamentous growth in moist nutrient-rich intestines of healthy arthropods and similar habitats. PMID:9448315

Margulis, L; Jorgensen, J Z; Dolan, S; Kolchinsky, R; Rainey, F A; Lo, S C

1998-02-01

372

The Arthromitus stage of Bacillus cereus: intestinal symbionts of animals  

NASA Technical Reports Server (NTRS)

In the guts of more than 25 species of arthropods we observed filaments containing refractile inclusions previously discovered and named "Arthromitus" in 1849 by Joseph Leidy [Leidy, J. (1849) Proc. Acad. Nat. Sci. Philadelphia 4, 225-233]. We cultivated these microbes from boiled intestines of 10 different species of surface-cleaned soil insects and isopod crustaceans. Literature review and these observations lead us to conclude that Arthromitus are spore-forming, variably motile, cultivable bacilli. As long rod-shaped bacteria, they lose their flagella, attach by fibers or fuzz to the intestinal epithelium, grow filamentously, and sporulate from their distal ends. When these organisms are incubated in culture, their life history stages are accelerated by light and inhibited by anoxia. Characterization of new Arthromitus isolates from digestive tracts of common sow bugs (Porcellio scaber), roaches (Gromphodorhina portentosa, Blaberus giganteus) and termites (Cryptotermes brevis, Kalotermes flavicollis) identifies these flagellated, spore-forming symbionts as a Bacillus sp. Complete sequencing of the 16S rRNA gene from four isolates (two sow bug, one hissing roach, one death's head roach) confirms these as the low-G+C Gram-positive eubacterium Bacillus cereus. We suggest that B. cereus and its close relatives, easily isolated from soil and grown on nutrient agar, enjoy filamentous growth in moist nutrient-rich intestines of healthy arthropods and similar habitats.

Margulis, L.; Jorgensen, J. Z.; Dolan, S.; Kolchinsky, R.; Rainey, F. A.; Lo, S. C.

1998-01-01

373

Bacillus cereus var. toyoi promotes growth, affects the histological organization and microbiota of the intestinal mucosa in rainbow trout fingerlings.  

PubMed

In this preliminary study, we evaluated the effects of a gram-positive soil bacteria Bacillus cereus var. toyoi on the growth performance, digestive enzyme activities, intestinal morphology, and microbiota in rainbow trout Oncorhynchus mykiss fingerlings. Trout were maintained in a recirculation system and fed 2 diets: 1) a commercial trout feed deprived of the probiotic and 2) the same diet but with the spores of the probiotic bacteria dissolved in fish oil during the manufacturing of the feed (final concentration = 2 × 10(4) cfu/g). Each diet was tested in three 400-L cylindroconical tanks (125 fish per tank; initial density = 1.3 kg/m(3); 13.2°C) for a period of 93 d. The probiotic-supplemented diet promoted growth, and the final mean BW and standard length in fish fed the probiotic were 3.4% and 2.1%, respectively, which was greater than the control group (P < 0.05). Fish fed the probiotic showed a more homogeneous distribution in the final BW, with a greater frequency of individuals around the modal of the normal distribution of the population. This result is of practical importance because homogenous production lots can improve rearing practices, reducing hierarchical dominance situations arising from individuals of larger sizes. In addition, the probiotic-supplemented diet increased the level of leukocyte infiltration in the lamina propria of the intestinal mucosa, the number of goblet cells (P < 0.010), and villi height (P < 0.001) but did not affect villi width. The administration of the probiotic changed the intestinal microbiota as indicated by 16S rDNA PCR-restriction fragment length polymorphism. In this sense, fish fed the probiotic formed a well-defined cluster composed of 1 super clade, whereas compared control fish had a greater degree of diversity in their gut microbiota. These changes in gut microbiota did not affect the specific activity of selected pancreatic and intestinal digestive enzymes. These results indicate that the inclusion of the probiotic bacteria in trout feeds could be beneficial for the host by enhancing its intestinal innate immune function and promoting growth. PMID:23508031

Gisbert, E; Castillo, M; Skalli, A; Andree, K B; Badiola, I

2013-06-01

374

Antibacterial effect (in vitro) of Moringa oleifera and Annona muricata against Gram positive and Gram negative bacteria.  

PubMed

Antibacterial effects of aqueous and ethanolic extracts of seeds of moringa (Moringa oleifera) and pods of soursop (Annona muricata) in the concentration of 1:5 and 1:10 in volumes 50, 100, 150 and 200 microL were examined against Staphylococcus aureus, Vibrio cholerae, Escherichia coli (isolated from the organism and the aquatic environment) and Salmonella Enteritidis. Antibacterial activity (inhibition halo > 13 mm) against S. aureus, V. cholerae and E. coli isolated from the whiteleg shrimp, Litopenaeus vannmaei, was detected in aqueous and ethanolic extracts of moringa. E. coli isolated from tilapiafish, Oreochromis niloticus, was sensitive to the ethanolic extract of moringa. The aqueous extracts of soursop showed an antibacterial effect against S. aureus and V. cholerae, but the antibacterial activity by the ethanol extracts of this plant was not demonstrated. PMID:20602021

Viera, Gustavo Hitzschky Fernandes; Mourão, Jozeanne Alves; Angelo, Angela Maria; Costa, Renata Albuquerque; Vieira, Regine Helena Silva dos Fernandes

2010-01-01

375

Organization and evolution of the cotG and cotH genes of Bacillus subtilis.  

PubMed

The cotG and cotH genes of Bacillus subtilis encode two previously characterized spore coat proteins. The two genes are adjacent on the chromosome and divergently transcribed by ?(K), a sporulation-specific ? factor of the RNA polymerase. We report evidence that the cotH promoter maps 812 bp upstream of the beginning of its coding region and that the divergent cotG gene is entirely contained between the promoter and the coding part of cotH. A bioinformatic analysis of all entirely sequenced prokaryotic genomes showed that such chromosomal organization is not common in spore-forming bacilli. Indeed, CotG is present only in B. subtilis, B. amyloliquefaciens, and B. atrophaeus and in two Geobacillus strains. When present, cotG always encodes a modular protein composed of tandem repeats and is always close to but divergently transcribed with respect to cotH. Bioinformatic and phylogenic data suggest that such genomic organizations have a common evolutionary origin and that the modular structure of the extant cotG genes is the outcome of multiple rounds of gene elongation events of an ancestral minigene. PMID:21984783

Giglio, Rosa; Fani, Renato; Isticato, Rachele; De Felice, Maurilio; Ricca, Ezio; Baccigalupi, Loredana

2011-12-01

376

Direct screening of recombinants in gram-positive bacteria using the secreted staphylococcal nuclease as a reporter.  

PubMed

A system for direct screening of recombinant clones in Lactococcus lactis, based on secretion of the staphylococcal nuclease (SNase) in the organism, was developed. The nuc gene (encoding SNase) was cloned on both rolling-circle and theta-replicating plasmids. L. lactis strains containing these nuc+ plasmids secrete SNase and are readily detectable by a simple plate test. A multicloning site (MCS) was introduced just after the cleavage site between leader peptide and the mature SNase, without affecting nuclease activity. Cloning foreign DNA fragments into any site of the MCS interrupts nuc and thus results in nuc mutant clones which are easily distinguished fron nuc+ clones on plates. The utility of this system for L. lactis was demonstrated by cloning an antibiotic resistance marker and Escherichia coli chromosomal DNA fragments into the MCS of the nucMCS cassette. Both cloning vectors containing the nucMCS cassette were also introduced into Streptococcus salivarius subsp. thermophilus, in which direct screening of nuc mutant recombinant clones was also achieved. The potential uses of nuc as a secretion reporter system are discussed. PMID:8051029

Le Loir, Y; Gruss, A; Ehrlich, S D; Langella, P

1994-08-01

377

Modulation of Connexin Expression and Gap Junction Communication in Astrocytes by the Gram-Positive Bacterium S. aureus  

PubMed Central

Gap junctions establish direct intercellular conduits between adjacent cells and are formed by the hexameric organization of protein subunits called connexins (Cx). It is unknown whether the proinflammatory milieu that ensues during CNS infection with S. aureus, one of the main etiologic agents of brain abscess in humans, is capable of eliciting regional changes in astrocyte homocellular gap junction communication (GJC) and, by extension, influencing neuron homeostasis at sites distant from the primary focus of infection. Here we investigated the effects of S. aureus and its cell wall product peptidoglycan (PGN) on Cx43, Cx30, and Cx26 expression, the main Cx isoforms found in astrocytes. Both bacterial stimuli led to a time-dependent decrease in Cx43 and Cx30 expression; however, Cx26 levels were elevated following bacterial exposure. Functional examination of dye coupling, as revealed by single-cell microinjections of Lucifer yellow, demonstrated that both S. aureus and PGN inhibited astrocyte GJC. Inhibition of protein synthesis with cyclohexamide (CHX) revealed that S. aureus directly modulates, in part, Cx43 and Cx30 expression, whereas Cx26 levels appear to be regulated by a factor(s) that requires de novo protein production; however, CHX did not alter the inhibitory effects of S. aureus on astrocyte GJC. The p38 MAPK inhibitor SB202190 was capable of partially restoring the S. aureus-mediated decrease in astrocyte GJC to that of unstimulated cells, suggesting the involvement of p38 MAPK-dependent pathway(s). These findings could have important implications for limiting the long-term detrimental effects of abscess formation in the brain which may include seizures and cognitive deficits.

ESEN, NILUFER; SHUFFIELD, DEBBIE; MOHSIN, MD. SYED; KIELIAN, TAMMY

2008-01-01

378

Toxin-Antitoxin Genes of the Gram-Positive Pathogen Streptococcus pneumoniae: So Few and Yet So Many  

PubMed Central

Summary: Pneumococcal infections cause up to 2 million deaths annually and raise a large economic burden and thus constitute an important threat to mankind. Because of the increase in the antibiotic resistance of Streptococcus pneumoniae clinical isolates, there is an urgent need to find new antimicrobial approaches to triumph over pneumococcal infections. Toxin-antitoxin (TA) systems (TAS), which are present in most living bacteria but not in eukaryotes, have been proposed as an effective strategy to combat bacterial infections. Type II TAS comprise a stable toxin and a labile antitoxin that form an innocuous TA complex under normal conditions. Under stress conditions, TA synthesis will be triggered, resulting in the degradation of the labile antitoxin and the release of the toxin protein, which would poison the host cells. The three functional chromosomal TAS from S. pneumoniae that have been studied as well as their molecular characteristics are discussed in detail in this review. Furthermore, a meticulous bioinformatics search has been performed for 48 pneumococcal genomes that are found in public databases, and more putative TAS, homologous to well-characterized ones, have been revealed. Strikingly, several unusual putative TAS, in terms of components and genetic organizations previously not envisaged, have been discovered and are further discussed. Previously, we reported a novel finding in which a unique pneumococcal DNA signature, the BOX element, affected the regulation of the pneumococcal yefM-yoeB TAS. This BOX element has also been found in some of the other pneumococcal TAS. In this review, we also discuss possible relationships between some of the pneumococcal TAS with pathogenicity, competence, biofilm formation, persistence, and an interesting phenomenon called bistability.

Chan, Wai Ting; Moreno-Cordoba, Inma

2012-01-01

379

Antibacterial activity of silver-doped hydroxyapatite nanoparticles against gram-positive and gram-negative bacteria  

PubMed Central

Ag-doped nanocrystalline hydroxyapatite nanoparticles (Ag:HAp-NPs) (Ca10-xAgx(PO4)6(OH)2, xAg?=?0.05, 0.2, and 0.3) with antibacterial properties are of great interest in the development of new products. Coprecipitation method is a promising route for obtaining nanocrystalline Ag:HAp with antibacterial properties. X-ray diffraction identified HAp as an unique crystalline phase in each sample. The calculated lattice constants of a?=?b?=?9.435 Å, c?=?6.876 Å for xAg?=?0.05, a?=?b?=?9.443 Å, c?=?6.875 Å for xAg?=?0.2, and a?=?b?=?9.445 Å, c?=?6.877 Å for xAg?=?0.3 are in good agreement with the standard of a?=?b?=?9.418 Å, c?=?6.884 Å (space group P63/m). The Fourier transform infrared and Raman spectra of the sintered HAp show the absorption bands characteristic to hydroxyapatite. The Ag:HAp nanoparticles are evaluated for their antibacterial activity against Staphylococcus aureus, Klebsiella pneumoniae, Providencia stuartii, Citrobacter freundii and Serratia marcescens. The results showed that the antibacterial activity of these materials, regardless of the sample types, was greatest against S. aureus, K. pneumoniae, P. stuartii, and C. freundii. The results of qualitative antibacterial tests revealed that the tested Ag:HAp-NPs had an important inhibitory activity on P. stuartii and C. freundii. The absorbance values measured at 490 nm of the P. stuartii and C. freundii in the presence of Ag:HAp-NPs decreased compared with those of organic solvent used (DMSO) for all the samples (xAg?=?0.05, 0.2, and 0.3). Antibacterial activity increased with the increase of xAg in the samples. The Ag:HAp-NP concentration had little influence on the bacterial growth (P. stuartii).

2012-01-01

380

Localization and Interactions of Teichoic Acid Synthetic Enzymes in Bacillus subtilis  

Microsoft Academic Search

The thick wall of gram-positive bacteria is a polymer meshwork composed predominantly of peptidoglycan (PG) and teichoic acids, both of which have a critical function in maintenance of the structural integrity and the shape of the cell. In Bacillus subtilis 168 the major teichoic acid is covalently coupled to PG and is known as wall teichoic acid (WTA). Recently, PG

Rut Carballido-López; Alex Formstone; Dirk-Jan Scheffers; Philippe Noirot; Jeffery Errington

2008-01-01

381

Draft Genome Sequence of Bacillus thuringiensis NBIN-866 with High Nematocidal Activity.  

PubMed

Bacillus thuringiensis NBIN-866, a Gram-positive bacterium, was isolated from soil in China. We announce here the draft genome sequence of strain B. thuringiensis NBIN-866, which possesses highly nematocidal factors, such as proteins and small molecular peptides. PMID:24855295

Liu, Xiaoyan; Zhou, Ronghua; Fu, Guiping; Zhang, Wei; Min, Yong; Tian, Yuxi; Huang, Daye; Wang, Kaimei; Wan, Zhongyi; Yao, Jingwu; Yang, Ziwen

2014-01-01

382

Myeloid Cell Sirtuin-1 Expression Does Not Alter Host Immune Responses to Gram-Negative Endotoxemia or Gram-Positive Bacterial Infection  

PubMed Central

The role of sirtuin-1 (SIRT1) in innate immunity, and in particular the influence of SIRT1 on antimicrobial defense against infection, has yet to be reported but is important to define since SIRT1 inhibitors are being investigated as therapeutic agents in the treatment of cancer, Huntington’s disease, and autoimmune diseases. Given the therapeutic potential of SIRT1 suppression, we sought to characterize the role of SIRT1 in host defense. Utilizing both pharmacologic methods and a genetic knockout, we demonstrate that SIRT1 expression has little influence on macrophage and neutrophil antimicrobial functions. Myeloid SIRT1 expression does not change mortality in gram-negative toxin-induced shock or gram-positive bacteremia, suggesting that therapeutic suppression of SIRT1 may be done safely without suppression of myeloid cell-specific immune responses to severe bacterial infections.

Crotty Alexander, Laura E.; Marsh, Brenda J.; Timmer, Anjuli M.; Lin, Ann E.; Zainabadi, Kayvan; Czopik, Agnieszka; Guarente, Leonard; Nizet, Victor

2013-01-01

383

A new hybrid bacteriocin, Ent35-MccV, displays antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria  

PubMed Central

Bacteriocins and microcins are ribosomally synthesized antimicrobial peptides that are usually active against phylogenetically related bacteria. Thus, bacteriocins are active against Gram-positive while microcins are active against Gram-negative bacteria. The narrow spectrum of action generally displayed by bacteriocins from lactic acid bacteria represents an important limitation for the application of these peptides as clinical drugs or as food biopreservatives. The present study describes the design and expression of a novel recombinant hybrid peptide combining enterocin CRL35 and microcin V named Ent35–MccV. The chimerical bacteriocin displayed antimicrobial activity against enterohemorrhagic Escherichia coli and Listeria monocytogenes clinical isolates, among other pathogenic bacteria. Therefore, Ent35–MccV may find important applications in food or pharmaceutical industries.

Acuna, Leonardo; Picariello, Gianluca; Sesma, Fernando; Morero, Roberto D.; Bellomio, Augusto

2012-01-01

384

Gram-positive bacterial lipoglycans based on a glycosylated diacylglycerol lipid anchor are microbe-associated molecular patterns recognized by TLR2.  

PubMed

Innate immune recognition is the first line of host defense against invading microorganisms. It is a based on the detection, by pattern recognition receptors (PRRs), of invariant molecular signatures that are unique to microorganisms. TLR2 is a PRR that plays a major role in the detection of Gram-positive bacteria by recognizing cell envelope lipid-linked polymers, also called macroamphiphiles, such as lipoproteins, lipoteichoic acids and mycobacterial lipoglycans. These microbe-associated molecular patterns (MAMPs) display a structure based on a lipid anchor, being either an acylated cysteine, a glycosylated diacylglycerol or a mannosyl-phosphatidylinositol respectively, and having in common a diacylglyceryl moiety. A fourth class of macroamphiphile, namely lipoglycans, whose lipid anchor is made, as for lipoteichoic acids, of a glycosylated diacylglycerol unit rather than a mannosyl-phosphatidylinositol, is found in Gram-positive bacteria and produced by certain Actinobacteria, including Micrococcus luteus, Stomatococcus mucilaginosus and Corynebacterium glutamicum. We report here that these alternative lipoglycans are also recognized by TLR2 and that they stimulate TLR2-dependant cytokine production, including IL-8, TNF-? and IL-6, and cell surface co-stimulatory molecule CD40 expression by a human macrophage cell line. However, they differ by their co-receptor requirement and the magnitude of the innate immune response they elicit. M. luteus and S. mucilaginosus lipoglycans require TLR1 for recognition by TLR2 and induce stronger responses than C. glutamicum lipoglycan, sensing of which by TLR2 is dependent on TLR6. These results expand the repertoire of MAMPs recognized by TLR2 to lipoglycans based on a glycosylated diacylglycerol lipid anchor and reinforce the paradigm that macroamphiphiles based on such an anchor, including lipoteichoic acids and alternative lipoglycans, induce TLR2-dependant innate immune responses. PMID:24278450

Blanc, Landry; Castanier, Romain; Mishra, Arun K; Ray, Aurélie; Besra, Gurdyal S; Sutcliffe, Iain; Vercellone, Alain; Nigou, Jérôme

2013-01-01

385

Accuracy and potential usefulness of triplex real-time PCR for improving antibiotic treatment of patients with blood cultures showing clustered gram-positive cocci on direct smears.  

PubMed

Bacterial identification and antibiotic susceptibility testing currently require 48 h when a first blood culture (BC) is positive for clustered gram-positive cocci on direct smear examination (DSE). Meanwhile, antibiotic treatment is often inadequate, reducing the chances of effective treatment or creating unnecessary selective pressure. A new real-time PCR (RT-PCR) technique that differentiates Staphylococcus aureus from coagulase-negative staphylococci (CoNS) and detects methicillin resistance in 90 min in BC bottles could help solve these problems. BC bottles from 410 patients with gram-positive cocci on DSE were processed by current methods, and patients' treatments were prospectively recorded. The RT-PCR assay was performed on aliquots of these BCs, which had been kept frozen. For the 121 patients who had true bacteremia, we established whether the faster availability of RT-PCR results could have led to the initiation of treatments different from those actually given. RT-PCR sensitivity and specificity were 100% for differentiating between S. aureus and CoNS and detecting methicillin resistance with two manufacturers' BC bottles. For 31/86 (36%) of the S. aureus-infected patients and for 8/35 (23%) of the CoNS-infected patients who either had suboptimal or nonoptimal treatment or were untreated 48 h after positivity was detected, the early availability of RT-PCR results could have allowed more effective treatment. Unnecessary glycopeptide treatments could have been avoided for 28 additional patients. The use of RT-PCR would increase treatment effectiveness in patients with staphylococcal bacteremia and reduce the selective pressure created by glycopeptides. PMID:18417663

Ruimy, Raymond; Dos-Santos, Marie; Raskine, Laurent; Bert, Frédéric; Masson, René; Elbaz, Sandrine; Bonnal, Christine; Lucet, Jean-Christophe; Lefort, Agnès; Fantin, Bruno; Wolff, Michel; Hornstein, Michele; Andremont, Antoine

2008-06-01

386

Bacillus rhizosphaerae sp. nov., an novel diazotrophic bacterium isolated from sugarcane rhizosphere soil  

Microsoft Academic Search

A Gram-positive, non-pigmented, rod-shaped, diazotrophic bacterial strain, designated SC-N012T, was isolated from rhizosphere soil of sugarcane and was subjected to a polyphasic taxonomic study. The strain exhibited\\u000a phenotypic properties that included chemotaxonomic characteristics consistent with its classification in the genus Bacillus. Sequence analysis of the 16S rRNA gene of SC-N012T revealed the closest match (98.9% pair wise similarity) with Bacillus

Munusamy Madhaiyan; Selvaraj Poonguzhali; Jung-Sook Lee; Keun-Chul Lee; Kuppusamy Hari

387

Identification of Tn 4430 , a transposon of Bacillus thuringiensis functional in Escherichia coli  

Microsoft Academic Search

The mobile genetic element Tn4430, originating from the gram-positive bacterium, Bacillus thuringiensis, and previously described as the Th-sequence, is the first transposon isolated from the genus Bacillus. In the present work a gene (APH-III) conferring resistance to kanamycin was inserted into this 4.2 kb transposon. Transposition experiments showed that Tn4430OAPH-III could transpose in the gram-negative host Escherichia coli when its

Didier Lereclus; Jacques Mahillon; Ghislaine Menou; Marguerite-M. Lecadet

1986-01-01

388

Risk assessment and ecological effects of transgenic Bacillus thuringiensis crops on non-target organisms.  

PubMed

The application of recombinant DNA technology has resulted in many insect-resistant varieties by genetic engineering (GE). Crops expressing Cry toxins derived from Bacillus thuringiensis (Bt) have been planted worldwide, and are an effective tool for pest control. However, one ecological concern regarding the potential effects of insect-resistant GE plants on non-target organisms (NTOs) has been continually debated. In the present study, we briefly summarize the data regarding the development and commercial use of transgenic Bt varieties, elaborate on the procedure and methods for assessing the non-target effects of insect-resistant GE plants, and synthetically analyze the related research results, mostly those published between 2005 and 2010. A mass of laboratory and field studies have shown that the currently available Bt crops have no direct detrimental effects on NTOs due to their narrow spectrum of activity, and Bt crops are increasing the abundance of some beneficial insects and improving the natural control of specific pests. The use of Bt crops, such as Bt maize and Bt cotton, results in significant reductions of insecticide application and clear benefits on the environment and farmer health. Consequently, Bt crops can be a useful component of integrated pest management systems to protect the crop from targeted pests. PMID:21564541

Yu, Hui-Lin; Li, Yun-He; Wu, Kong-Ming

2011-07-01

389

Co-production of surfactin and a novel bacteriocin by Bacillus subtilis subsp. subtilis H4 isolated from Bikalga, an African alkaline Hibiscus sabdariffa seed fermented condiment.  

PubMed

Bikalga is a Hibiscus sabdariffa seed fermented condiment widely consumed in Burkina Faso and neighboring countries. The fermentation is dominated by Bacillus subtilis group species. Ten B. subtilis subsp. subtilis (six isolates) and Bacillus licheniformis (four isolates) isolated from traditional Bikalga were examined for their antimicrobial activity against a panel of 36 indicator organisms including Gram-positive and Gram-negative bacteria and yeasts. The Bacillus spp. isolates showed variable inhibitory abilities depending on the method used. Both Gram-positive and Gram-negative bacteria were inhibited in the agar spot assay while only Gram-positive pathogens were inhibited in the agar well diffusion assay. Cell free supernatants (CFS) of pure cultures of 3 B. subtilis subsp. subtilis (G2, H4 and F1) strains inhibited growth of Listeria monocytogenes, Micrococcus luteus, Staphylococcus aureus and Bacillus cereus, while CFS of 2 B. licheniformis (E3 and F9) strains only inhibited M. luteus. The antimicrobial substance(s) produced by B. subtilis subsp. subtilis H4 was further characterized. The antimicrobial substance(s) produced by H4 was detected from mid-exponential growth phase. The activity was sensitive to protease and trypsin, but resistant to the proteolytic action of proteinase K and papain. Treatment with ?-amylase and lipase II resulted in a complete loss of antimicrobial effect, indicating that a sugar moiety and lipid moiety are necessary for the activity. Treatment with mercapto-ethanol resulted in a significant loss, indicative of the presence of disulfide bridges. The antimicrobial activity of H4 was heat resistant and active at pH3-10. PCR detection of yiwB, sboA, spoX, albA and spaS, etnS genes and genes coding for surfactins and plipastatins (fengycins) indicated a potential for subtilosin, subtilin and lipopeptide production, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was carried out and a single band of approximately 4kDa had antimicrobial activity. Ultra high performance liquid chromatography-time of flight mass spectrometry (UHPLC-TOFMS) analysis of the 4kDa band allowed identification of surfactin and a protein with a monoisotopic mass of 3346.59Da, which is dissimilar in size to subtilosin and subtilin. Surfactin is a cyclic lipoheptapeptide, which contains a ?-hydroxy fatty acid, but no di-sulfide bridges or sugar residues. The complete loss of activity upon amylase treatment indicates that surfactin was not responsible for the observed antimicrobial effect. However, it cannot completely be ruled out that surfactin acts synergistically with the detected protein, though further investigations are needed to confirm this. PMID:23466466

Compaoré, Clarisse S; Nielsen, Dennis S; Ouoba, Labia I I; Berner, Torben S; Nielsen, Kristian F; Sawadogo-Lingani, Hagrétou; Diawara, Bréhima; Ouédraogo, Georges A; Jakobsen, Mogens; Thorsen, Line

2013-04-01

390

Expression of low endotoxin 3-O-sulfotransferase in Bacillus subtilis and Bacillus megaterium.  

PubMed

A key enzyme for the biosynthesis and bioengineering of heparin, 3-O-sulfotransferase-1 (3-OST-1), was expressed and purified in Gram-positive Bacillus subtilis and Bacillus megaterium. Western blotting, protein sequence analysis, and enzyme activity measurement confirmed the expression. The enzymatic activity of 3-OST-1 expressed in Bacillus species were found to be similar to those found when expressed in Escherichia coli. The endotoxin level in 3-OST-1 from B. subtilis and B. megaterium were 10(4)-10(5)-fold lower than that of the E. coli-expressed 3-OST-1, which makes the Bacillus expression system of particular interest for the generation of pharmaceutical grade raw heparin from nonanimal sources. PMID:23912211

Wang, Wenya; Englaender, Jacob A; Xu, Peng; Mehta, Krunal K; Suwan, Jiraporn; Dordick, Jonathan S; Zhang, Fuming; Yuan, Qipeng; Linhardt, Robert J; Koffas, Mattheos

2013-10-01

391

The complete genome sequence of Bacillus licheniformis DSM13, an organism with great industrial potential  

Microsoft Academic Search

The genome of Bacillus licheniformis DSM13 consists of a single chromosome that has a size of 4,222,748 base pairs. The average G+C ratio is 46.2%. 4,286 open reading frames, 72 tRNA genes, 7 rRNA operons and 20 transposase genes were identified. The genome shows a marked co-linearity with Bacillus subtilis but contains defined inserted regions that can be identified at

Birgit Veith; Christina Herzberg; Silke Steckel; Jörg Feesche; Karl Heinz Maurer; Petra Ehrenreich; S. Baumer; Anke Henne; Heiko Liesegang; Rainer Merkl; Armin Ehrenreich; Gerhard Gottschalk

2004-01-01

392

Increased stability of an esterase from Bacillus stearothermophilus in ionic liquids as compared to organic solvents  

Microsoft Academic Search

Three different ionic liquids, 1-butyl-3-methyl imidazolium bis[(trifluoromethyl)sulfonyl]amide [BMIM][BTA], 1-butyl-3-methyl imidazolium hexafluorophosphate [BMIM][PF6] and 1-butyl-3-methyl imidazolium tetrafluoroborate [BMIM][BF4] were used as reaction media for the transesterification of 1-phenylethanol catalysed by esterases from Bacillus subtilis and Bacillus stearothermophilus. No transesterification activity could be detected in the ionic liquids when the lyophilised powder of the esterases was used as biocatalysts. By immobilising the

Mattias Persson; Uwe T. Bornscheuer

2003-01-01

393

Genome of a Gut Strain of Bacillus subtilis  

PubMed Central

Bacillus subtilis is a Gram-positive, rod-shaped, spore-forming bacterium. We present the genome sequence of an undomesticated strain, BSP1, isolated from poultry. The sequence of the BSP1 genome supports the view that B. subtilis has a biphasic lifestyle, cycling between the soil and the animal gastrointestinal tract, and it provides molecular-level insight into the adaptation of B. subtilis to life under laboratory conditions.

Serra, Claudia R.; Lapointe, Thomas; Pereira-Leal, Jose B.; Potot, Sebastien; Fickers, Patrick; Perkins, John B.; Wyss, Markus; Henriques, Adriano O.

2013-01-01

394

Meta-analysis of Randomized Controlled Trials of Vancomycin for the Treatment of Patients With Gram-Positive Infections: Focus on the Study Design  

PubMed Central

Objective To study the effectiveness and safety of vancomycin compared with that of other antibiotics for the treatment of gram-positive infections. Methods Major electronic databases were searched. Data from published randomized controlled trials (January 1, 1950, to September 15, 2011) were pooled using a meta-analytic method. Results Fifty-three trials comparing vancomycin with linezolid, daptomycin, quinupristin-dalfopristin, tigecycline, ceftaroline, ceftobiprole, telavancin, teicoplanin, iclaprim, and dalbavancin were included in the meta-analysis. Individual antibiotics were as effective as vancomycin, except for linezolid, which was more effective than vancomycin for the treatment of skin and soft tissue infections (odds ratio [OR], 1.61; 95% confidence interval [CI], 1.07-2.43). Comparators were as effective as vancomycin in the intent-to-treat population (OR, 1.08; 95% CI, 0.98-1.18) but were more effective in the clinically evaluable population (OR, 1.14; 95% CI, 1.02-1.27) when all infections were pooled. When available data from all trials were pooled, no differences were noted when patients with febrile neutropenia (OR, 1.07; 95% CI, 0.82-1.39), pneumonia (OR, 1.10; 95% CI, 0.87-1.37), bacteremia (OR, 1.05; 95% CI, 0.76-1.45), and skin and soft tissue infections (OR, 1.11; 95% CI, 0.89-1.39) were studied. Comparators were more effective in open-label (OR, 1.28; 95% CI, 1.08-1.50) but not in double-blind trials (OR, 1.04; 95% CI, 0.90-1.20). Total adverse events attributed to studied antibiotics (OR, 1.07; 95% CI, 0.90-1.28) and patients withdrawn from trials (OR, 0.86; 95% CI, 0.68-1.09) were similar in the compared groups. Mortality was not different between vancomycin and comparator antibiotics when all trials were included in the analysis (OR, 1.09; 95% CI, 0.96-1.23). Comparators were associated with higher mortality in open-label (OR, 1.27; 95% CI, 1.05-1.54) but not double-blind trials (OR, 0.96; 95% CI, 0.80-1.14). Conclusion On the basis mainly of data from open-label trials, vancomycin is a treatment choice that is as effective as other available antibiotics for patients with gram-positive infections. Study design seems to make a major contribution to the outcome.

Vardakas, Konstantinos Z.; Mavros, Michael N.; Roussos, Nikolaos; Falagas, Matthew E.

2012-01-01

395

Bacillus axarquiensis sp. nov. and Bacillus malacitensis sp. nov., isolated from river-mouth sediments in southern Spain  

Microsoft Academic Search

Two Gram-positive, rod-shaped, endospore-forming bacteria (strains CR-119T and CR-95T) were isolated from brackish sediments in the mouth of the river Velez in Malaga, southern Spain, and subjected to a polyphasic taxonomic study. Phenotypic tests showed that these strains were related to other Bacillus species at a similarity level of less than 87?6 %. Both strains are halotolerant, aerobic, chemoheterotrophic, motile

Cristina Ruiz-Garcia; Emilia Quesada; Fernando Martinez-Checa; Inmaculada Llamas; Maria C. Urdaci; Victoria Bejar

2005-01-01

396

Rapid identification and antimicrobial susceptibility testing of Gram-positive cocci in blood cultures by direct inoculation into the BD Phoenix system.  

PubMed

Rapid identification and antimicrobial susceptibility testing (AST) of the causative agent(s) of bloodstream infections are essential for the selection of appropriate antimicrobial therapy. To speed up the iden