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PubMed1992

Early diagnosis of infectious keratitis with in vivo real time confocal microscopy.

The tandem scanning confocal microscope (TSM) was adapted for in vivo examination of the cornea in rabbits with experimental bacterial and fungal keratitis. Compared to slit lamp biomicroscopy, the TSM provides superior lateral and axial resolution and serial optical sectioning capability, which may be useful for identification of corneal pathogens in the early stages of infection. We used the TSM to examine normal rabbit eyes infected with bacteria (Bacillus cereus) and a filamentous fungus (Aspergillus). We also examined a human cornea removed by penetrating keratoplasty after a clinical diagnosis of amoebic keratitis. In the early stages of bacterial infection, slit lamp examination revealed a nonspecific minimal stromal haze and limbal injection indistinguishable from sterile ulcers and epithelial defects. With the TSM, bacteria were visible as highly refractile bodies in the epithelium and superficial stroma. Branching fungal hyphae were also easily identified by the TSM, as were Acanthamoeba cysts and parasites in the subepithelial stroma. Our results indicate that this technique may provide a new modality for quickly and accurately identifying the agent of corneal infection, thereby facilitating prompt and appropriate treatment.

Acanthamoeba Keratitis↗
PubMed2004

[Purification and cloning of an antifungal protein from the rice diseases controlling bacterial strain Paenibacillus polymyxa WY110].

Paenibacillus polymyxa WY110, a plant growth-promoting bacteria strain isolated from rice rhizosphere could suppress the growth of various plant pathogens effectively. With (NH4)2SO4 fractional precipitation, DEAE-Sephadex A-50 chromatography and Sephacryl S-200 chromatography followed by tracks of fraction antagonistic assay and SDS-PAGE, an antifungal protein P2 with in vitro anti-Pyricularia oxyzae activity was isolated and purified. It was showed with antagonistic activity on PDA plates that the growth of Pyricularia oryzae was inhibited by 1.5 microg of P2 protein effectively. N-terminal amino acid residues analysis showed 24 amino acid sequence: H2N-Ala-Asn-Val-Phe-Trp-Glu-Pro-Leu-Ser-Tyr-Tyr-Asn-Pro-Ser-Thr-Trp-Gln-Lys-Ala-Asp-Gly-Tyr-Ser-Asn-. Using this amino acid sequence as a target, the similarity of P2 protein was searched with BlastP program on Internet. It was showed a high homology between the P2 protein and the precursors of beta-1, 3-1, 4-glucanases from Bacillus. The beta-1, 3-1, 4-glucanase activity of P2 protein was identified with the specific substrate lichenan. According to the N-terminal partial sequence of P2 protein and the C-terminal conserved sequence of beta-1, 3-1, 4-glucanase, the primers for both terminals were synthesized. Using the genomic DNA of WY110 as the template, the full-length sequence of the gene encoding P2 was amplified by high fidelity PCR, then cloned into pMD18-T vecter. Sequence analysis showed the 72 nucleotide sequence on 5'-end matched with the known 24 amino acid sequence on N-terminal of P2 protein. The sequence (GenBank Accession Number: AF284449) was 636 bp in length encoding 212 amino acids. Comparing with a beta-1, 3-1, 4-glucanase gene (gluB) from Paenibacillus polymyxa, the sequence homology for nucleotides and deduced amino acids were 84% and 88.7% respectively. The cloning of the gene encoding P2 protein would be a new potential objective gene for plant gene engineering.

Amino Acid Sequence↗
PubMed2005

The diversity of culturable organotrophic bacteria from local solar salterns.

We isolated and cultured bacteria inhabiting solar saltern ponds in Taean-Gun, Chungnam Province, Korea. All of the isolated 64 strains were found to be moderately halophilic bacteria, growing in a salt range of 2-20 %, with an optimal concentration of 5% salt. Bacterial diversity among the isolated halophiles was evaluated via RFLP analyses of PCR-amplified 16S rDNAs, followed by phylogenetic analysis of the partial 16S rDNA sequences. The combination of restriction enzyme digestions with HaeIII, CfoI, MspI and RsaI generated 54 distinct patterns. A neighbor-joining tree of the partial 16S rDNA sequences resulted in the division of the 64 strains into 2 major groups, 45 strains of gamma-Proteobacteria (70.3%) and 19 strains of Firmicutes (29.7%). The alpha-Proteobacteria and Cytophaga-Flavobacterium-Bacterioides groups, which were repeatedly found to exist in thalassohaline environments, were not represented in our isolates. The gamma-Proteobacteria group consisted of several subgroups of the Vibrionaceae (37.5%), Pseudoalteromonadaceae (10.9%), Halomonadaceae (7.8%), Alteromonadaceae (7.8%), and Idiomarinaceae (6.3%). Members of Salinivibrio costicola (29.7%) were the most predominant species among all of the isolates, followed by Halobacillus treperi (12.5%). Additionally, three new species candidates were found, based on similarities of the 16S rDNA sequences to those of previously published species.

Bacillaceae↗
PubMed2005

[Construction of the genomic library of Halobacillus sp. D8 and isolation of the glycine betaine transporter betH gene].

Halobacillus sp. D8 is a sporing-forming, gram-positive moderately halophilic bacterium which could tolerate up to 25% (W/V) NaCl. A genomic library of Halobacillus sp. D8 was constructed using pUC18 as vector, and 9000 recombinant plasmids were obtained. By dot blot hybridization, colony PCR and DNA sequencing, the entire glycine betaine transporter betH gene was isolated from the constructed library. Inspection of the sequenced 4.3 kb DNA region revealed the presence of three ORFs. The putative ORF of betH is 1515bp long, encoding a 505-residue protein (BetH) with a calculated molecular mass of 56.1kD. Hydrophobicity plot analysis of BetH indicated a transmembrane protein containing 12 transmembrane regions. Homology searches for BetH of strain D8 in the GenBank using the BLAST program revealed significant sequence identities to other glycine betaine transporters: the putative glycine betaine transporter of O. iheyensis (64% identity), OpuD of B. subtilis (51% identity), BetH of H. trueperi (49% identity), BetL of L. monocytogenes (48% identity), BetM of M. halophilus (43% identity) and the putative glycine betaine transporter of B. halodurans (44% identity).

Amino Acid Sequence↗
PubMed2004

[Amphibacillus haojiensis sp. nov.--a novel alkaliphilic and slight halophilic bacterium from Haoji Soda Lake in Inner Mongolia Autonomous Region, China].

An alkaliphilic and slightly halophilic Gram-positive rod, designated strain F10, was isolated from Haoji soda lake in Inner Mongolia Autonomous Region, China. Strain F10 grew optimally in the presence of 3% (W/V) NaCl, pH 9.5 and 37 degrees C. It was facultatively anaerobic, motile with peritrichous flagella, sporulating, catalase- negative. The (G + C) content of the DNA was 40 mol%. The cell wall contained meso-DAP. Phylogenetic analyses based on 16S rDNA sequences comparisons indicate that strain F10 was a member of the genus Amphibacillus. DNA-DNA similarity of less than 30% with the described species of Amphibacillus supported the view that this organism represented a new species of the genus Amphibacillus. The name Amphibacillus haojiensis sp. nov. is proposed.

Bacillaceae↗
PubMedDate not supplied

[Bacillus subtilis and phenotypically similar strains producing hexaene antibiotics].

We studied metabolites synthesized by Bacillus subtilis strains, including the type strain of B. atrophaeus and phenotypically similar cultures. These metabolites were presented by polyene antibiotics with conjugated double bonds. Hexaenes from study strains inhibited the growth of phytopathogenic fungi Fusarium culmorum, F. sporotrichiella, F. oxysporum, Botrytis sorokiniana, Alternaria tenuis, and Phytophtora infestans. The degree of growth inhibition depended on the test fungus.

Anti-Bacterial Agents↗
PubMed2005

[Screening and identification of steroidal saponins-producing endophytes from Paris polyphylla var. chinensis Franch].

Two endophytic strains SS01 and SS02 with the potential for producing steroidal saponins were isolated from the underground stems of Paris polyphylla var. chinensis Franch. The TLC comparison indicated that there are 3 sports with similar R(f) between the metabolites of SS01 and the saponins of Paris polyphylla var. chinensis Franch. And there are 2 sports with similar R(f) between the metabolites of SS02 and the saponins of Paris polyphylla var. chinensis The and that biochemical SS01 Franch. characteristics of morphology, physiological belonged showed to Enterobacteriaceae and SS02 belonged to Bacillus sp.. The 16S rDNA of SS01 and SS02 were PCR and sequenced. The accessions of GenBank are AY842143 and AY842144, respectively. The two 16S rDNA phylogenetic trees were constructed by comparing with the published 16S rDNA sequences of the relative bacteria species. In the first phylogenetic tree SS01 and Cedecea davisae DSM 4568 was the closest relative with 98.9% sequence similarity, and in the second phylogenetic tree SS02 and Paenibacillus daejeonensis was the closest relative with 97.7% sequence similarity. According to the phylogenetic analysis they were identified as Cedecea davisae SS01 and Paenibacillus daejeonensis SS02, respectively.

Bacillaceae↗
PubMed2005

[Randomised placebo controlled double blind study on the effect of prebiotic oligosaccharides on intestinal flora in healthy infants].

AIM: Effects of supplementing prebiotic oligosaccharides to formula for healthy infants were studied in this placebo controlled, randomised, double blind study. METHODS: Ninety-seven infants were included into the study; among them 42 breast-fed infants, 14 infants fed formula supplemented with 0.4 g/100ml oligosaccharides (9 to 1 mixture of galacto- and fructooligosaccharides) and 13 infants fed control formula were followed-up throughout the 12-week-long study period. The groups receiving formula were compared with statistical methods, whereas data of breast-fed infants served as reference values. RESULTS: Infants fed the two formulae did not differ in nutrient intakes, growth, occurrence rate of feeding difficulties and atopic manifestations, or in calcium excretion. The intestinal flora did not differ between the two formula fed groups at the beginning of the study. In contrast, numbers of Bifidobacteriae were significantly higher in infants receiving the formula supplemented with prebiotic oligosaccharides both at the 14th day (9 x 1011 versus 5 x 1010, colony forming units/g faeces, median, p < 0.05) and 28th day (9 x 1012 versus 5 x 1010, p < 0.05) of the study. CONCLUSION: In this study, supplementation of infant formula with prebiotic oligosaccharides resulted in ameliorating the difference in intestinal flora between formula fed and breast-fed healthy infants.

Bacillaceae↗
PubMed2006

Protein expression analysis of Halobacillus dabanensis D-8T subjected to salt shock.

To investigate the mechanism of salt tolerance of gram-positive moderately halophilic bacteria, two-dimensional gel electrophoresis (2-D PAGE) was employed to achieve high resolution maps of proteins of Halobacillus dabanensis D-8T. Approximately 700 spots of proteins were identified from these 2-D PAGE maps. The majority of these proteins had molecular weights between 17.5 and 66 kDa, and most of them were distributed between the isoelectric points (pI) 4.0 and 5.9. Some protein spots were distributed in the more acidic region of the 2-D gel (pI <4.0). This pattern indicated that a number of proteins in the strain D-8T are acidic. To understand the adaptation mechanisms of moderately halophilic bacteria in response to sudden environmental changes, differential protein profiles of this strain were investigated by 2-D PAGE and Imagemaster 2D Platinum software after the cells were subjected to salt shock of 1 to 25% salinity for 5 and 50 min. Analysis showed 59 proteins with an altered level of expression as the result of the exposure to salt shock. Eighteen proteins had increased expression, 8 proteins were induced, and the expression of 33 proteins was down-regulated. Eight of the up-regulated proteins were identified using MALDI-TOF/MS and MASCOT, and were similar to proteins involved in signal transduction, proteins participating in energy metabolism pathways and proteins involved in stress.

Bacillaceae↗
PubMed2006

[Construction of the shuttle expression-secretion vector with the promoters and signal peptide-encoding sequence from Brevibacillus brevis].

The multiple and tandem promoters and signal peptide-encoding sequence of cell wall protein encoding gene was amplified from Brevibacillus brevis B15 total DNA, the PCR fragment was cloned, sequenced and analyzed, then was submitted to GenBank with a Accession No. AY956423. Another pair of primers were designed to amplify the fragment again, BamHI and Pstl sites was introduced flanking the PCR production. BamHI/Pstl digested fragment was cloned into the corresponding site of shuttle vector pP43NMK to generate the expression-secretion vector pP15MK. The inserted fragment was upstream of mpd gene and the signal peptide-encoding sequence was fused in frame with the mpd gene, which its own signal peptide-encoding sequence was deleted. The recombinant vector was transformed into Bacillus subtilis 1A751, under the control of the promoters and signal peptide from Brevibacillus brevis B15, mpd gene was continuously expressed and secreted at a high efficiency throughout the exponential growth phase and into the late stationary phase, the expression production methyl parathion hydrolase (MPH) was attached on the outside of the cell membrane. MPH activity accumulated to a maximum level of 7.79 U/mL after 48 h of cultivation at the late stationary phase, which was 8.1-fold higher than the expression level of the original Plesiomonas strain M6.

Bacillaceae↗
PubMed2006

[Two-dimensional gel electrophoresis analysis of moderately halophilic bacterium Halobacillus dabanensis D-8T under hypoosmotic shock condition].

Halobacillus dabanensis D-8T was isolated from the saline deposits of Daban lake in Xinjiang of China, and is able to grow in complex medium containing 0.5% to 25% salt. To figure out the survival mechanisms of Gram-positive moderately halophilic bacteria under hypoosmotic shock conditions, two-dimensional gel electrophoresis (2-DE) was carried out to investigate differential protein expression profiles of H. dabanensis D-8T in response to low osmotic challenge. The 2-D gels were stored as dry gels and their images were taken by ImageScanner and analyzed by Imagemaster 2D Platinum software. About 650 protein spots were detected in 2-D gel. Most of proteins were distributed in molecular mass of 17.5 - 66kDa and the range of isoelectric point 4.0 - 5.9. A total of 34 protein spots were found to alter their expression after strain D-8T was subjected to hypoosmotic shock from 20% to 0% salinity for 5 min and 50 min. Among them, the expression of 20 protein spots is up-regulated including 6 new protein spots, while that of 14 protein spots is down-regulated in answer to sudden osmotic down-shift. Protein spots of interest were excised from the gels and digested by trypsin. By means of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) and MASCOT search engine, 4 up-regulated protein spots were identified with peptide mass fingerprint, and are similar to heat shock protein DanK, rod shape-determining protein, penicillin-binding protein (PBP-1A) and 5-enolpyruvoylshikimate-3-phosphate synthase, respectively. Noticeably, PBP-1A firstly was up-regulated after shock of 5 min but disappeared after shock of 50 min. This indicated that the strain activate a minor mechanism of peptidoglycan synthesis to compensate the major synthesis mechanism for cells survival through a down-shift challenge. In addition, this paper was the first report that heat shock proteins were up-regulated in response to sudden osmotic down-shift.

Bacillaceae↗
PubMed1991

Results in the management of Bacillus endophthalmitis.

Eighteen cases of culture-proven Bacillus endophthalmitis were reviewed. The majority (13/18) were secondary to penetrating ocular trauma. Others were secondary to corneal ulcer (1), postoperative (1), and endogenous (3). Bacillus cereus was the most commonly identified species (12/18). Antibiotic sensitivities demonstrated uniform sensitivity to vancomycin and the aminoglycosides. Visual prognosis was generally poor; only two patients maintained 20/200 or better in the affected eye. Fourteen eyes were enucleated or eviscerated. The remaining two eyes had light perception vision at the last follow-up examination.

Adult↗
PubMed1987

Novel phenylalanine dehydrogenases from Sporosarcina ureae and Bacillus sphaericus. Purification and characterization.

NAD+-dependent phenylalanine dehydrogenases were purified 1,500- and 1,600-fold, and crystallized from Sporosarcina ureae SCRC-R04 and Bacillus sphaericus SCRC-R79a, respectively. The purified enzymes were homogeneous as judged by disc gel electrophoresis. The enzyme from S. ureae has a molecular weight of 305,000, while that of B. sphaericus has a molecular weight of 340,000. Each is probably composed of eight subunits identical in molecular weight. The S. ureae enzyme showed a high substrate specificity in the oxidative deamination reaction acting on L-phenylalanine, while that of B. sphaericus acted on L-phenylalanine and L-tyrosine. The enzymes had lower substrate specificities in the reductive amination reaction acting on alpha-keto acids. The Sporosarcina enzyme acted on phenylpyruvate, alpha-ketocaproate, alpha-keto-gamma-methylthiobutyrate and rho-hydroxyphenylpyruvate. The Bacillus enzyme acted on rho-hydroxyphenylpyruvate, phenylpyruvate, and alpha-keto-gamma-methylthiobutyrate. The enzyme from B. sphaericus catalyzes The enzyme from B. sphaericus catalyzes the transfer of pro-S (B) hydrogen from NADH.

Amino Acid Oxidoreductases↗
PubMed1978

A randomized prospective evaluation of orally administered antibiotics in operations on the colon.

In this randomized, prospective study of 91 patients undergoing operations on the colon, we found that neomycin, neomycin-phthalylsulfathiazole and neomycin-erythromycin base all provided adequate intestinal antisepsis. Wound infection rates for all groups were also similar and close to the national average for operations on the colon. A detailed analysis of the wound infections in our series revealed that 60 per cent of these infections were from organisms different from those found on the colonic mucosa. This evidence indicates the possible advantage of parenteral antibiotic therapy to achieve adequate tissue levels of antibiotics not presently possible with orally administered agents.

Administration, Oral↗
PubMed1987

[Effect of components of curing mixtures on the antagonistic properties of enterococci and other bacteria isolated from cured meat].

Effect of sodium chloride, glucose, sodium glutamate, potassium nitrate, sodium nitrate, sodium nitraite, ascorbic acid, sodium citrate, polyphosphates (Hamine) on antagonistic activity of enterococci and various bacteria isolated from cured meat against Bacillus cereus, Bacillus subtilis, Salmonella typhi-murium, Salmonella cholerae-suis, Staphylococcus aureus, Clostridium botulinum type A, Clostridium botulinum type B, Clostridium botulinum type E, Clostridium bifermentans, Clostridium perfringens type A, Clostridium sporogenes. It was found that some of the chemical compounds examined increased the antagonistic effect and other compounds partly or totally inhibited it. Sodium nitrate, sodium nitrite, potassium nitrate, glucose, and ascorbic acid in majority of cases increased antagonism and sodium citrate and polyphosphates restrained this phenomenon.

Animals↗
PubMedDate not supplied

[Fixation of molecular nitrogen by sulfate-reducing bacteria from petroleum strata].

The activity of nitrogen fixation by the museum strains of sulfate reducing bacteria isolated from oil deposits was studied using the acetylene technique. The mesophilic sulpfate reducing bacteria belonging to the species Desulfovibrio africanus 2372 and D. baculatus X were found to have a high nitrogenase activity. D desulfuricans subsp. aestuarii 2198 reduced acetylene at a low rate. The thermophilic sulfate reducing cultures of D. thermophilus 7, Desulfotomaculum nigrificans 781 and Dm. nigrificans subsp. salinus 435 produced only small quantities of ethylene. Apparently, nitrogen fixation by sulfate reducing bacteria can be considerable in oil strata whose temperature does not exceed 35--40 degrees C.

Bacillaceae↗