Microb Pathog, 2004 Jul, 37(1), 1 - 9 Effect of W07-toxin on gut physiological response in mice; Bhattacharyya S et al.; A number of unknown secretogenic factor(s) from Vibrio cholerae have been implicated to play a role in inducing cholera-like symptoms observed in patients . The present study has been carried out on the novel W07-toxin (pI 5.2) from V . cholerae W07, an epidemic cholera strain devoid of the ctx gene . The toxin showed maximum binding to GM(1) and interacted with a 20 kDa glycoprotein present on the cell membrane of mice enterocytes in a GM(1) specific manner . The analysis of biochemical parameters in enterocytes triggered with this toxin revealed a significant increase in intracellular calcium concentration and a massive secretion of Cl(-) . However, no absorption of Na(+) was observed under the same condition . This toxin also elevated the level of cyclic adenosine 3',5'-monophosphate (cAMP) as well as protein kinase A (PKA) . Thus, the novel toxin, although distinct from cholera-toxin, showed some functional homology to it and may be one of the key players inducing electrolyte imbalance within intestinal cells in the cholera-like symptoms associated with V . cholerae W07. Vaccine, 2004 Jun 23, 22(19), 2457 - 69 Biosafety aspects of the recombinant live oral Vibrio cholerae vaccine strain CVD 103-HgR; Viret JF et al.; The development of live attenuated vaccines, allowing for the safe and effective immunisation at mucosal surfaces, is a strategy of great interest for vaccinologists . The main advantage of this approach over conventional parenteral vaccines is the induction of strong mucosal immune responses, allowing targeting of the pathogen at the initial point of contact with the host . Further advantages include the ease of administration, high acceptance by vaccines, and relatively low production costs . Finally, well-characterised, safe and immunogenic vaccine strains are well suited as vectors for the mucosal delivery of foreign vaccine antigens and of DNA vaccines . However, such vaccines, when based on or containing genetically modified organisms (GMOs), are facing new and specific regulatory hurdles, particularly regarding the potential risks for humans and the environment . In this contribution we address selected aspects of the risk assessment of live attenuated bacterial vaccines covered in the course of the registration of vaccine strain CVD 103-HgR as a recombinant live oral vaccine against cholera. J Fish Dis, 2004 Jun, 27(6), 319 - 26 Competition for attachment of aquaculture candidate probiotic and pathogenic bacteria on fish intestinal mucus; Vine NG et al.; Probiotics for aquaculture are generally only selected by their ability to produce antimicrobial metabolites; however, attachment to intestinal mucus is important in order to remain within the gut of its host . Five candidate probiotics (AP1-AP5), isolated from the clownfish, Amphiprion percula (Lacepede), were examined for their ability to attach to fish intestinal mucus and compete with two pathogens, Aeromonas hydrophila and Vibrio alginolyticus . Two different radioactive isotopes were used to quantify competition between pathogens and probionts . Attachment of the pathogens was enhanced by the presence of the candidate probiotics . However, the addition of the candidate probiotics after the pathogens resulted in reduced pathogen attachment . Only AP5 caused lower attachment success of V . alginolyticus when added before the pathogen . When AP5 was added first, the average attachment change was 41% compared with 72% when added after V . alginolyticus, suggesting that the probiotic is displaced but that enhanced attachment of the pathogen does not occur . Conversely, when V . alginolyticus was added first, followed by AP5, attachment change was 37% while AP5 had 92% attachment change when added second . This implies that the pathogen was displaced by the candidate probiotic and therefore it appeared that, based on the ability of probiont AP5 to attach to mucus, the growth of the pathogen in the digestive tract might be suppressed by the candidate probiont's presence. Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 11 - 4 {Triacylglycerol lipase activity of hemolytic Vibrio cholerae}; Telesmanich NR; The new method for the determination of the lipase activity of purified preparations of hemolysins and live V . cholerae cell has been developed . On the basis of the determination of triacylglycerollipase (lipase) activity the test for the differentiation of hemolytic and nonhemolytic V . cholerae has been proposed. Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 7 - 10 {Transition of Vibrio cholerae into the non-culturable state under the conditions of low temperature, different mineralization and illumination of artificial culture medium}; Nikoleishvili LR et al.; The importance of the combined influence of temperature, mineralization and illumination of the medium on the time of the transition of V . cholerae into the uncultivable state has been shown . The reversion of 5- to 60-day variants of uncultivable forms after the elevation of temperature to 20-22 degrees C has been obtained. Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 3 - 7 {Characterization of Vibrio cholerae O1 cultures isolated from environmental objects on the territory of the Russian Federation in 2002}; Onishchenko GG et al.; The biological properties of 46 V . cholerae O1 eltor cultures isolated in 2002 from water environment on the territory of Russia are presented . All isolated vibrios proved to be typical in their cultural, morphological, biochemical and serological properties . The atypical character of some of them was mainly linked with their phage resistance . The appearance of vibrios, sensitive to bacteriophage ctx+ and containing gene tcp in the absence gene ctx, was noted . Multilocus VNTR typing made it possible to group the cultures under study in 34 genotypes . The presence of toxin coregulated pili was found to be directly related to locus VcB . The necessity of the systematic study of the pheno- and genotypes of the isolated cultures with the aim of epidemiological surveillance of this infection is emphasized. Proteomics, 2004 May, 4(5), 1491 - 504 A proteome reference map for Vibrio cholerae El Tor; Coelho A et al.; A proteome reference map has been constructed for Vibrio cholerae El Tor, in the pI range of 4.0 to 7.0 . The map is based on two-dimensional gels (2-D) and the identification, by peptide mass fingerprint, of proteins in 94 spots, corresponding to 80 abundant proteins . Two strains are compared, strain N16961 and a Latin American El Tor strain C3294 . The consensus map contains 340 spots consistently seen with both strains grown in Luria-Bertani broth (LB) or minimal M9 medium . The results were obtained from nine gels run with 18 cm immobilized pH gradient strips and precast gels . The 2-D gels were anchored to real N16961 proteins identified by mass spectrometry . Various energy metabolism components and periplasmic ATP-binding cassette (ABC) transporter proteins were identified among the abundant proteins . Two isoforms of OmpU were found . Five operons are proposed and seven hypothetical proteins were experimentally confirmed . Comparisons are made with protein 2-D gels for a classical strain and to microarray analysis available for the N16961 El Tor strain . New results were obtained from the proteome analysis, indicating an abundance of periplasmic ABC transporter proteins not found in microarray studies. Mol Microbiol, 2004 Jun, 52(6), 1677 - 89 A distinctive dual-channel quorum-sensing system operates in Vibrio anguillarum; Croxatto A et al.; Many bacterial cells communicate using diffusible signal molecules to monitor cell population density via a process termed quorum sensing . In marine Vibrio species, the Vibrio harveyi-type LuxR protein is a key player in a quorum-sensing phosphorelay cascade, which controls the expression of virulence, symbiotic and survival genes . Previously, we characterized Vibrio anguillarum homologues of LuxR (VanT) and LuxMN (VanMN) and, in this study, we have identified homologues of LuxPQ (VanPQ) and LuxOU (VanOU) . In contrast to other Vibrio species, vanT was expressed at low cell density and showed no significant induction as the cell number increased . In addition, although the loss of VanO increased vanT expression, the loss of VanU, unexpectedly, decreased it . Both VanN and VanQ were required for repression of vanT even in a vanU mutant, suggesting an alternative route for VanNQ signal transduction other than via VanU . VanT negatively regulated its own expression by binding and repressing the vanT promoter and by binding and activating the vanOU promoter . The signal relay results in a cellular response as expression of the metalloprotease, empA, was altered similar to that of vanT in all the mutants . Consequently, the V . anguillarum quorum-sensing phosphorelay systems work differently from those of V . harveyi and may be used to limit rather than induce vanT expression. Environ Microbiol, 2004 Jul, 6(7), 760 - 3 Polylysogeny and prophage induction by secondary infection in Vibrio cholerae; Espeland EM et al.; Strains of Vibrio cholerae O1, biotypes El Tor and classical, were infected with a known temperate phage (PhiP15) and monitored over a 15-day period for prophage induction . Over the course of the experiment two morphologically and three genomically distinct virus-like particles were observed from the phage-infected El Tor strain by transmission electron microscopy and field inversion gel electrophoresis, respectively, whereas only one phage, PhiP15, was observed from the infected classical strain . In the uninfected El Tor culture one prophage was spontaneously induced after 6 days . No induction in either strain was observed after treatment with mitomycin C . Data indicate that El Tor biotypes of V . cholerae may be polylysogenic and that secondary infection can promote multiple prophage induction . These traits may be important in the transfer of genetic material among V . cholerae by providing an environmentally relevant route for multiple prophage propagation and transmission. Environ Microbiol, 2004 Jul, 6(7), 699 - 706 Occurrence and distribution of Vibrio cholerae in the coastal environment of Peru; Gil AI et al.; The occurrence and distribution of Vibrio cholerae in sea water and plankton along the coast of Peru were studied from October 1997 to June 2000, and included the 1997-98 El Nino event . Samples were collected at four sites in coastal waters off Peru at monthly intervals . Of 178 samples collected and tested, V . cholerae O1 was cultured from 10 (5.6%) samples, and V . cholerae O1 was detected by direct fluorescent antibody assay in 26 out of 159 samples tested (16.4%) . Based on the number of cholera cases reported in Peru from 1997 to 2000, a significant correlation was observed between cholera incidence and elevated sea surface temperature (SST) along the coast of Peru (P < 0.001) . From the results of this study, coastal sea water and zooplankton are concluded to be a reservoir for V . cholerae in Peru . The climate-cholera relationship observed for the 1997-98 El Nino year suggests that an early warning system for cholera risk can be established for Peru and neighbouring Latin American countries. Microbiology, 2004 Jun, 150(Pt 6), 1769 - 77 Molecular identification of Vibrio harveyi-related isolates associated with diseased aquatic organisms; Gomez-Gil B et al.; Fifty strains belonging to Vibrio harveyi, Vibrio campbellii, and the recently described Vibrio rotiferianus, were analysed using phenotypic and genomic techniques with the aim of analysing the usefulness of the different techniques for the identification of V . harveyi-related species . The species V . harveyi and V . campbellii were phenotypically indistinguishable by more than 100 phenotypic features . Thirty-nine experimental strains were phenotypically identified as V . harveyi, but FAFLP, REP-PCR, IGS-PCR and DNA-DNA hybridization proved that they in fact belong to the species V . campbellii . Similar groupings were found among all fingerprinting methodologies (except IGS-PCR) . Thirty-two experimental strains clustered with the V . campbellii type and one reference strain; seven strains clustered with the V . harveyi type and three reference strains; and the type and four reference strains of V . rotiferianus grouped together . The correlations between DNA-DNA hybridization and the genomic fingerprinting by FAFLP and (GTG)(5)-PCR were found to be above 0.68 and statistically significant, suggesting the value of the latter techniques for the reliable identification of V . harveyi-related species . The results presented indicate that strains phenotypically identified as V . harveyi are in fact V . campbellii; these findings position V . campbellii as an important species involved in diseases of reared aquatic organisms. FEMS Microbiol Lett, 2004 Jun 15, 235(2), 243 - 8 A plasmid-encoded class 1 integron carrying sat, a putative phosphoserine phosphatase gene and aadA2 from enterotoxigenic Escherichia coli O159 isolated in Japan; Ahmed AM et al.; A class 1 integron was detected in a single multidrug-resistant strain of enterotoxigenice Escherichia coli (ETEC) O159 after examination of 23 clinical E . coli isolates . This isolate was resistant to streptomycin, kanamycin, gentamicin, chloramphenicol and ampicillin . Sequencing of the class 1 integron identified three-gene cassettes . The first is the streptothricin acetyltransferase gene, sat, which confers resistance to streptothricin . The second is an ORF whose product is a putative phosphoserine phosphatase (PSP), and the last is an aminoglycoside adenyltransferase gene, aadA2, which confers resistance to streptomycin and spectinomycin . The putative PSP gene product was found to be 39%, 38%, 28%, and 27% identical to PSP gene products of Vibrio vulnificus CMCP6, V . vulnificus YJ016, Pseudomonas syringae, and P . aeruginosa, respectively . Southern-blot hybridization showed that this integron is located on a 90 kb plasmid . This is the first report identifying a putative PSP gene in an integron. Zhonghua Yu Fang Yi Xue Za Zhi, 2004 May, 38(3), 200 - 3 {Characteristics of virulence gene in Vibrio parahaemolyticus strains isolated from clinical patients and environment in Hangzhou, China}; Zhang W et al.; OBJECTIVES: To investigate the characteristics of virulence gene in Vibrio parahaemolyticus strains isolated from clinical patients and environment in Hangzhou, China . METHODS: Thermostable direct hemolysin gene (tdh) and thermostable direct hemolysin-related hemolysin gene (trh) were determined in a total of 174 strains of V . parahaemolyticus isolated from patients and environment (seafood) in Hangzhou area by PCR . RESULTS: The tdh was found in 92 out of 94 V . parahaemolyticus strains from food poisoning patients and in 33 out of 34 strains from sporadic diarrhea patients, and trh was not detected in all above clinical strains . Meanwhile the tdh was negative in all V . parahaemolyticus strains from environment, and the trh was also negative except one strain with urease activity . All strains with trh negative had no the activity of urease . CONCLUSIONS: The V . parahaemolyticus strains from food poisoning patients and sporadic diarrhea patients are tdh positive and trh negative . The V . parahaemolyticus strains with tdh negative and almost trh positive in environment might be a potential pathogen in Hangzhou. J Infect Dis, 2004 Jun 15, 189(12), 2318 - 22 Epub 2004 May 25. Incomplete correlation of serum vibriocidal antibody titer with protection from Vibrio cholerae infection in urban Bangladesh; Saha D et al.; The serum vibriocidal antibody is the only recognized predictor of protection from cholera, but no seroepidemiological data have been gathered since the emergence of Vibrio cholerae O139 . We assessed the association between the vibriocidal antibody titer and protection from cholera in an endemic setting . Although a higher baseline vibriocidal titer correlated with protection from V . cholerae O1, infection still developed in some contacts with very high titers . No association between baseline vibriocidal titer and protection from V . cholerae O139 infection was found . Our findings suggest that the vibriocidal antibody is an incomplete predictor of protection from V . cholerae infection. Protein Expr Purif, 2004 Jul, 36(1), 115 - 23 Expression, purification, and sequence analysis of catalase-1 from the soil bacterium Comamonas terrigena N3H; Zamocky M et al.; Catalases are essential components of the cellular equipment to cope with oxidative stress . We have purified and characterize herein the most abundant heme-containing catalase-1 from the soil bacterium Comamonas terrigena N3H . This oxidative stress-induced enzyme was isolated from exponential phase cells grown in the presence of peroxyacetic acid . We have used consecutive steps of hydrophobic, molecular sieve, and ion exchange chromatography to achieve a high state of purity for this metalloenzyme . The purified sample of catalase exhibited a specific catalytic activity of 55,900 U/mg, allosteric behavior in peroxidic reaction, a broad pH optimum, and a rather atypical electronic spectrum . The sample of highest purity was subjected to mass spectrometry analysis . The molecular weight of the subunit of this homodimeric protein was determined as 55,417 Da . The Qq-TOF mass analysis method allowed us to sequence short tryptic fragments of this catalase . Five such fragments with a total length of 57 amino acids together with several enzymatic properties allowed the classification of this hydroperoxidase as belonging to clade III of monofunctional catalases . The highest sequence similarity is with the catalase from Vibrio fischeri . The presented results imply the significance of this inducible enzyme in the prevention of toxic effects of oxidative stress for bacterial cells. J Bacteriol, 2004 Jun, 186(12), 4014 - 8 Cross-regulation in Vibrio parahaemolyticus: compensatory activation of polar flagellar genes by the lateral flagellar regulator LafK; Kim YK et al.; Gene organization and hierarchical regulation of the polar flagellar genes of Vibrio parahaemolyticus, Vibrio cholerae, and Pseudomonas aeruginosa appear highly similar, with one puzzling difference . Two sigma(54)-dependent regulators are required to direct different classes of intermediate flagellar gene expression in V . cholerae and P . aeruginosa, whereas the V . parahaemolyticus homolog of one of these regulators, FlaK, appears dispensable . Here we demonstrate that there is compensatory activation of polar flagellar genes by the lateral flagellar regulator LafK. J Bacteriol, 2004 Jun, 186(12), 3873 - 81 Vibrio fischeri LuxS and AinS: comparative study of two signal synthases; Lupp C et al.; Vibrio fischeri possesses two acyl-homoserine lactone quorum-sensing systems, ain and lux, both of which are involved in the regulation of luminescence gene expression and are required for persistent colonization of the squid host, Euprymna scolopes . We have previously demonstrated that the ain system induces luminescence at cell densities that precede lux system activation . Our data suggested that the ain system both relieves repression and initially induces the lux system, thereby achieving sequential induction of gene expression by these two systems . Analysis of the V . fischeri genome revealed the presence of a putative third system based on the enzyme LuxS, which catalyzes the synthesis of the Vibrio harveyi autoinducer 2 (AI-2) . In this study, we investigated the impact of V . fischeri LuxS on luminescence and colonization competence in comparison to that of the ain system . Similar to the ain system, inactivation of the AI-2 system decreased light production in culture, but not in the squid host . However, while an ainS mutant produces no detectable light in culture, a luxS mutant expressed approximately 70% of wild-type luminescence levels . A mutation in luxS alone did not compromise symbiotic competence of V . fischeri; however, levels of colonization of an ainS luxS double mutant were reduced to 50% of the already diminished level of ainS mutant colonization, suggesting that these two systems regulate colonization gene expression synergistically through a common pathway . Introduction of a luxO mutation into the luxS and ainS luxS background could relieve both luminescence and colonization defects, consistent with a model in which LuxS, like AinS, regulates gene expression through LuxO . Furthermore, while luxS transcription appeared to be constitutive and the AI-2 signal concentration did not change dramatically, our data suggest that ainS transcription is autoregulated, resulting in an over 2,000-fold increase in signal concentration as culture density increased . Taken together, these data indicate that V . fischeri LuxS affects both luminescence regulation and colonization competence; however, its quantitative contribution is small when compared to that of the AinS signal. J Bacteriol, 2004 Jun, 186(12), 3794 - 805 Quorum sensing regulates type III secretion in Vibrio harveyi and Vibrio parahaemolyticus; Henke JM et al.; In a process known as quorum sensing, bacteria communicate with one another by producing, releasing, detecting, and responding to signal molecules called autoinducers . Vibrio harveyi, a marine pathogen, uses two parallel quorum-sensing circuits, each consisting of an autoinducer-sensor pair, to control the expression of genes required for bioluminescence and a number of other target genes . Genetic screens designed to discover autoinducer-regulated targets in V . harveyi have revealed genes encoding components of a putative type III secretion (TTS) system . Using transcriptional reporter fusions and TTS protein localization studies, we show that the TTS system is indeed functional in V . harveyi and that expression of the genes encoding the secretion machinery requires an intact quorum-sensing signal transduction cascade . The newly completed genome of the closely related marine bacterium Vibrio parahaemolyticus, which is a human pathogen, shows that it possesses the genes encoding both of the V . harveyi-like quorum-sensing signaling circuits and that it also has a TTS system similar to that of V . harveyi . We show that quorum sensing regulates TTS in V . parahaemolyticus . Previous reports connecting quorum sensing to TTS in enterohemorrhagic and enteropathogenic Escherichia coli show that quorum sensing activates TTS at high cell density . Surprisingly, we find that at high cell density (in the presence of autoinducers), quorum sensing represses TTS in V . harveyi and V . parahaemolyticus. Genetika, 2004 Apr, 40(4), 445 - 53 {Role of moderate bacteriophage 139 in change in production of cholera toxin in a classic strain of Vibrio cholerae}; Eroshenko GA et al.; New data were obtained concerning cell sensitivity of pathogenic strains of cholera vibrions, which belong to the serogroup O1 of classical biovar, to the temperate bacteriophage K139, the native host of which is Vibrio cholerae O139 . Molecular-genetic and biochemical studies showed that phage 139 integrated into the chromosome of strains V . cholerae O1 can change their toxigenic properties . A change in the production of cholera toxin (CT) in lysogens is associated both with an increase in the activity of the toxR regulatory gene and with a distortion of the structure of a chromosomal DNA region that contains a copy of the operon ctxAB encoding CT biosynthesis. Mar Pollut Bull, 2004 Jun, 48(11-12), 1096 - 101 Multiplex PCR allows simultaneous detection of pathogens in ships' ballast water; Aridgides LJ et al.; There is enormous potential for global transfer of microorganisms, including pathogens, in ships' ballast water . We contend that a major advancement in the study of ballast-water microorganisms in particular, and of aquatic pathogens in general, will be expedited sample analysis, such as provided by the elegant technology of DNA microarrays . In order to use DNA microarrays, however, one must establish the appropriate conditions to bind target sequences in samples to multiple probes on the microarrays . We conducted proof-of-concept experiments to optimize simultaneous detection of multiple microorganisms using polymerase chain reaction (PCR) and Southern hybridization . We chose three target organisms, all potentially found in ballast water: a calicivirus, the bacterium Vibrio cholerae, and the photosynthetic protist Aureococcus anophagefferens . Here, we show simultaneous detection of multiple pathogens is possible, a result supporting the promising future use of microarrays for simultaneous detection of pathogens in ballast water. J Mol Microbiol Biotechnol, 2004, 7(1-2), 18 - 29 Dual flagellar systems enable motility under different circumstances; McCarter LL; Flagella are extremely effective organelles of locomotion used by a variety of bacteria and archaea . Some bacteria, including Aeromonas, Azospirillum, Rhodospirillum, and Vibrio species, possess dual flagellar systems that are suited for movement under different circumstances . Swimming in liquid is promoted by a single polar flagellum . Swarming over surfaces or in viscous environments is enabled by the production of numerous peritrichous, or lateral, flagella . The polar flagellum is produced continuously, while the lateral flagella are produced under conditions that disable polar flagellar function . Thus at times, two types of flagellar organelles are assembled simultaneously . This review focuses on the polar and lateral flagellar systems of Vibrio parahaemolyticus . Approximately 50 polar and 40 lateral flagellar genes have been identified encoding distinct structural, motor, export/assembly, and regulatory elements . The sodium motive force drives polar flagellar rotation, and the proton motive force powers lateral translocation . Polar genes are found exclusively on the large chromosome, and lateral genes reside entirely on the small chromosome of the organism . The timing of gene expression corresponds to the temporal demand for components during assembly of the organelle: RpoN and lateral- and polar-specific sigma(54)-dependent transcription factors control early/intermediate gene transcription; lateral- and polar-specific sigma(28) factors direct late flagellar gene expression . Although a different gene set encodes each flagellar system, the constituents of a central navigation system (i.e., chemotaxis signal transduction) are shared . Shokuhin Eiseigaku Zasshi, 2004 Feb, 45(1), 35 - 7 Growth kinetics of Vibrio parahaemolyticus O3:K6 under varying conditions of pH, NaCl concentration and temperature; Nishina T et al.; The growth responses of Vibrio parahamolyticus to pH, NaCl concentration and temperature changes were studied using serotype O3:K6 and other strains . Growth curves were obtained for 27 different sets of conditions, comprised of three levels of NaCl concentration, pH and temperature . The temperature, pH and NaCl concentrations most favorable for growth were in the order of 25 degrees C, 20 degrees C and 15 degrees C, pH 8, 7 and 5.8, and 1%, 3% and 7%, respectively . The bacteria grew most rapidly at 25 degrees C, at a pH of 7 or 8 in the presence of 1% or 3% NaCl, with the population (initial, ca . 2.5 log CFU/mL) reaching a level log 7 CFU/mL at 12 h . A growth predictive model using the Gompertz equation was generated from the experimental data for any combination of NaCl concentration, pH and temperature within the range used in this study. Mol Gen Mikrobiol Virusol, 2004, (2), 11 - 6 {A comparative analysis of genomes of virulent and avirulent strains of Vibrio cholerae O139}; Eroshenko GA et al.; A comparative analysis of the genome of V . cholerae O139 strains isolated in Russia's territory from patients with cholera and from the environment showed essential differences in their structures . The genome of clinical strains possessed all tested genes associated with virulence (ctxAB, zot, ace, rstC, rtxA, hap, toxR and toxT) and the at-tRS site for the CTXp phage DNA integration . As for the O139 V . cholerae chromosome strains isolated from water, 70% of the studied genes (ctxAB, zot, ace, rstC, tcpA, and toxT) and the attRS sequence were not detected in them . A lack of the key virulence genes in O139-serogroup "water" vibrios, including genes of toxin-coregulated adhesion pili . (that are receptors for the CTXp phage), and of the attachment site of the above phage are indicative of that the O139 V . cholerae strains isolated from open water sources located in different Russia's regions are epidemically negligible. Microbiol Res, 2004, 159(1), 19 - 28 Yeast DNA sequences initiating gene expression in Escherichia coli; Lewin A et al.; DNA transfer between pro- and eukaryotes occurs either during natural horizontal gene transfer or as a result of the employment of gene technology . We analysed the capacity of DNA sequences from a eukaryotic donor organism (Saccharomyces cerevisiae) to serve as promoter region in a prokaryotic recipient (Escherichia coli) by creating fusions between promoterless luxAB genes from Vibrio harveyi and random DNA sequences from S . cerevisiae and measuring the luminescence of transformed E . coli . Fifty-four out of 100 randomly analysed S . cerevisiae DNA sequences caused considerable gene expression in E . coli . Determination of transcription start sites within six selected yeast sequences in E . coli confirmed the existence of bacterial -10 and -35 consensus sequences at appropriate distances upstream from transcription initiation sites . Our results demonstrate that the probability of transcription of transferred eukaryotic DNA in bacteria is extremely high and does not require the insertion of the transferred DNA behind a promoter of the recipient genome. Microbes Infect, 2004 Jun, 6(7), 676 - 85 Association of adherence and motility in interleukin 8 induction in human intestinal epithelial cells by Vibrio cholerae; Sarkar M et al.; Interleukin 8 (IL-8) mRNA expression in Vibrio cholerae-infected human intestinal epithelial cells Int407 was determined by quantitative real-time RT-PCR and secretion measured by ELISA . Incubation of Int407 with V . cholerae O395 resulted in increased IL-8 mRNA expression as early as within 2 h of infection . Kinetics of IL-8 secretion reached a peak at about 8 h (780 pg/ml) and decreased thereafter . Induction of IL-8 was significantly high among various toxin-producing strains of V . cholerae belonging to serovar O1, O139 and non-O1 compared to non-toxinogenic strains . Induction of IL-8 was maximum in V . cholerae O395, required live cells and was dependent on de novo protein synthesis . The bacterial culture supernatant and crude cell envelope showed IL-8 stimulating activity . Infection of the monolayer with V . cholerae O395 cheY4 null mutant (O395YN), defective in adherence and motility, resulted in highly reduced levels of IL-8 expression, while hyperadherent and hypermotile mutant (O395Y) with the cheY4 gene duplicated also showed very high IL-8 expression . Another hyperadherent icmF insertion mutant (O395F) with reduced motility showed almost half the amount of IL-8 expression compared to O395Y . These results clearly indicate that both motility and adherence to intestinal epithelial cells are possible triggering factors contributing to IL-8 mRNA expression by V . cholerae. Biochem Biophys Res Commun, 2004 Jun 18, 319(1), 207 - 13 Bright fluorescence of a novel protein from Vibrio vulnificus depends on NADPH and the expression of this protein is regulated by a LysR-type regulatory gene; Chang CC et al.; A blue fluorescent protein, BfgV, belonging to the short-chain dehydrogenase/reductase superfamily, was found in a non-bioluminescent pathogen Vibrio vulnificus CKM-1 . This protein has fluorescence spectra with two excitation peaks at 283 and 352 nm, and one emission peak at 456 nm . BfgV fluoresces through effectively augmenting the intrinsic fluorescence of NADPH bound to it . Escherichia coli transformants expressing this protein can emit eye-detectable fluorescence . A LysR-type transcriptional regulator gene bfgR was found at the vicinal upstream region of bfgV in CKM-1 genome . The clues that products of bfgR can specifically bind to bfgR-bfgV intergenic promoter region and the deletion of bfgR significantly decreases the expression of bfgV reveal bfgR is a repressor gene of bfgV in V . vulnificus CKM-1. FEMS Microbiol Lett, 2004 Jun 1, 235(1), 199 - 207 Transcriptional analysis and operon structure of the tagA-orf2-orf3-mop-tagD region on the Vibrio pathogenicity island in epidemic V . cholerae; Zhang D et al.; The Vibrio pathogenicity island (VPI) in epidemic Vibrio cholerae is an essential virulence gene cluster . The VPI can excise from the chromosome and form extrachromosomal circular excision products . The VPI is 41.2-kb in size and encodes 29 potential proteins, several of which have no known function and whose regulation is not well understood . To determine the transcriptional organization of the tagA-orf2-orf3-mop-tagD region located at the 5'-(left) end of the VPI, we used reverse-transcriptase-PCR (RT-PCR), Northern blot analysis and DNA sequencing . RT-PCR primers were designed to transcribe and amplify regions spanning two or more open reading frames so as to establish the transcriptional organization . RT-PCR and Northern blot results demonstrated that the tagA-tagD region is transcribed as a polycistronic message and organized into several potential operons including tagA-orf2, orf3-mop, orf3-mop-tagD and tagD alone . Transcriptional lacZ fusions supported the existence of a promoter upstream of orf3 that was toxT-dependent . Interestingly, our data suggests that the orf3 promoter can drive the expression of either a long transcript (orf3-mop-tagD) or a short transcript (orf3-mop) without tagD . Our data also suggests that tagD can be expressed from two different promoters and that tagD is either transcribed alone or co-expressed with orf3-mop under certain conditions . These studies provide new insight into the genetic structure, transcriptional organization and regulation of a cluster of virulence genes on the VPI of epidemic V . cholerae. J Food Prot, 2004 May, 67(5), 1005 - 8 Real-time PCR detection of the thermostable direct hemolysin and thermolabile hemolysin genes in a Vibrio parahaemolyticus cultured from mussels and mussel homogenate associated with a foodborne outbreak; Davis CR et al.; Molecular methods have become vital epidemiological tools in the detection and characterization of bacteria associated with a foodborne outbreak . We used both culture and real-time PCR to detect a Vibrio parahaemolyticus isolate associated with a foodborne outbreak . The outbreak occurred in July 2002 in Polk County, Florida, and originated at a Chinese buffet, with one person being hospitalized . The hospital isolated V . parahaemolyticus from the patient but destroyed the sample after diagnosis . From an onsite visit of the restaurant, food samples that possibly contributed to the outbreak were collected and sent to the Florida Department of Health, Tampa Branch Laboratory . Crab legs, crabsticks, and mussel samples were homogenized and incubated according to the Food and Drug Administration Bacteriological Analytical Manual culture protocol . Three sets of primers and a TaqMan probe were designed to target the tdh, trh, and tlh genes and used for real-time PCR . This study was successful in isolating V . parahaemolyticus from a mussel sample and detecting two of its genes (tdh and tlh) in food and pure culture by real-time PCR. J Bacteriol, 2004 Jun, 186(11), 3304 - 12 Isolation and characterization of rpoS from a pathogenic bacterium, Vibrio vulnificus: role of sigmaS in survival of exponential-phase cells under oxidative stress; Park KJ et al.; A gene homologous to rpoS was cloned from a fatal human pathogen, Vibrio vulnificus . The functional role of rpoS in V . vulnificus was accessed by using an rpoS knockout mutant strain . This mutant was impaired in terms of the ability to survive under oxidative stress, nutrient starvation, UV irradiation, or acidic conditions . The increased susceptibility of the V . vulnificus mutant in the exponential phase to H2O2 was attributed to the reduced activity of hydroperoxidase I (HPI) . Although sigmaS synthesis was induced and HPI activity reached the maximal level in the stationary phase, the mutant in the stationary phase showed the same susceptibility to H2O2 as the wild-type strain in the stationary phase . In addition, HPII activity, which is known to be controlled by sigmaS in Escherichia coli, was not detectable in V . vulnificus strains under the conditions tested . The mutant in the exponential phase complemented with multiple copies of either the rpoS or katG gene of V . vulnificus recovered both resistance to H2O2 and HPI activity compared with the control strain . Expression of the katG gene encoding HPI in V . vulnificus was monitored by using a katG::luxAB transcriptional fusion . The expression of this gene was significantly reduced by deletion of sigmaS in both the early exponential and late stationary phases . Thus, sigmaS is necessary for increased synthesis and activity of HPI, and sigmaS is required for exponentially growing V . vulnificus to develop the ability to survive in the presence of H2O2. Vaccine, 2004 Jun 2, 22(17-18), 2137 - 45 Experimental immunisation and protection of guinea pigs with Vibrio cholerae toxoid and mucinases, neuraminidase and proteinase; Stewart-Tull DE et al.; As measured by fluid accumulation in ileal loops, Vibrio cholerae mucinase complex, with or without toxoid, protected guinea pigs from challenges with V . cholerae live organisms and enterotoxin . The neuraminidase and proteinases of the complex were combined in modified oil emulsion or aluminum hydroxide adjuvants and the resultant vaccines given by the parenteral or oral routes . There was little difference between the two types of adjuvant . Control of stomach acidity improved oral vaccination . Animals injected intramuscularly (i.m.) with toxoid-containing vaccines were protected from challenge with cholera toxin (CT) whereas those given oral doses were not . Toxoid plus killed V . cholerae cells elicited a more effective protection against toxin challenge than killed V . cholerae cells alone . Vaccines containing mucinases, with or without toxoid, protected the animals from a live V . cholerae challenge . The anti-mucinase immune response may prevent adhesion of the V . cholerae cells and hence reduce delivery of toxin to receptors . These mucinases, neuraminidase and proteinases, may be useful components of acellular, toxoided cholera vaccines for human immunisation. Genes Cells, 2004 May, 9(5), 479 - 95 Identification of cryptochrome DASH from vertebrates; Daiyasu H et al.; A new type of cryptochrome, CRY-DASH, has been recently identified . The CRY-DASH proteins constitute the fifth subfamily of the photolyase/cryptochrome family . CRY-DASHs have been identified from Synechocystis sp . PCC 6803, Vibrio cholerae, and Arabidopsis thaliana . The Synechocystis CRY-DASH was the first cryptochrome identified from bacteria, and its biochemical features and tertiary structure have been extensively investigated . To determine how broadly the subfamily is distributed within living organisms, we searched for new CRY-DASH candidates within several databases . We found five sequences as new CRY-DASH candidates, which are derived from four marine bacteria and Neurospora crassa . We also found many CRY-DASH candidates from the EST databases, which included sequences from fish and amphibians . We cloned and sequenced the cDNAs of the zebrafish and Xenopus laevis candidates, based on the EST sequences . The proteins encoded by the two genes were purified and characterized . Both proteins contained folate and flavin cofactors, and have a weak DNA photolyase activity . A phylogenetic analysis revealed that the seven candidates actually belong to the new type of cryptochrome subfamily . This is the first report of the CRY-DASH members from vertebrates and fungi. FEMS Immunol Med Microbiol, 2004 Jun 1, 41(2), 169 - 76 Regulation of proinflammatory mediator production in RAW264.7 macrophage by Vibrio vulnificus luxS and smcR; Shin NR et al.; Vibrio vulnificus causes fatal septicemia in human hosts, which is the consequence of raw shellfish consumption . The mortality following septicemia is dependent on the in vivo production of inflammatory mediators, including tumor necrosis factor-alpha (TNFalpha) . The present study was set up to investigate the association of quorum sensing in V . vulnificus with the host immune response . The effect of quorum sensing on cytotoxicity and the production of proinflammatory mediators was examined using the murine macrophage cell-line RAW264.7 . Cytotoxicity was determined by measuring lactate dehydrogenase release in the culture medium . Extracellular products from luxS- and smcR-deficient mutants exhibited weak cytotoxic effects on RAW264.7 cells . The production of the proinflammatory cytokines TNFalpha, IL-1beta and IL-6 was measured with real-time PCR and ELISA, and production was measured with Griess reagents . Mutation of both luxS and smcR delayed the transcription of TNFalpha, IL-1beta and IL-6 genes . Also, levels of both TNFalpha and nitric oxide induced by luxS- and smcR-deficient mutants were significantly lower than those induced by parent strains . These results suggest that quorum sensing could be involved in the modulation of TNFalpha and nitric oxide produced from host cells by regulating virulence factors, and that V . vulnificus facilitates its host's mortality and bacterial survival by enhancing virulence on host cells. Fish Shellfish Immunol, 2004 Jul, 17(1), 41 - 51 The immune stimulatory effect of sodium alginate on the white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus; Cheng W et al.; The total haemocyte count (THC), differential haemocyte count (DHC), phenoloxidase activity, respiratory burst (release of superoxide anion), superoxide dismutase activity, phagocytic activity and clearance efficiency to the pathogen Vibrio alginolyticus were measured when the white shrimp Litopenaeus vannamei (9.4-11.3 g) were injected individually with sodium alginate at 10, 20 or 50 microg g(-1) . No significant differences in THC, DHC and superoxide dismutase activity were observed among the shrimp injected with saline and those injected with sodium alginate at 10, 20 or 50 microg g(-1) . However, L . vannamei injected with sodium alginate at 20 microg g(-1)increased its phenoloxidase activity and respiratory burst after 2 days and one day, respectively . L . vannamei injected with sodium alginate at 50 microg g(-1)maintained a higher phagocytic activity and clearance efficiency to V . alginolyticus after 4 days . In another experiment, L . vannamei which had been injected with sodium alginate, were challenged with V . alginolyticus at 2x10(5)colony-forming units (CFU) shrimp(-1)and then placed in seawater of 34 per thousand . The survival of shrimp that received sodium alginate at either dose was significantly higher than that of control shrimp at the termination of the experiment (6 days after the challenge) . It is therefore concluded that L . vannamei received sodium alginate at 10 microg g(-1)or more and increased its immune ability and resistance from V . alginolyticus infection. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 2004 Apr, 21(2), 255 - 8 {Cloning and expression of immunoadjuvant molecule--CTB gene}; Wang T et al.; We cloned cholera toxin subunit B gene from 569B and M045 strain of Vibrio cholerae with polymerase chain reaction, constructed recombinant plasmid pCTB, and transformed pCTB into the prokaryotic cell strain JM109 . The indentification was made by means of restriction enzyme analysis, polymerase chain reaction, DNA sequencing, SDS-polyacrylamine gel electrophoresis analysis and Western blot . The results indicate that we have amplified cholera toxin subunit B gene of 376 bp from Vibrio cholerae and hve constructed the recombinant plasmid pCTB, and we have affained the object amied at successful expression of 12 KD in the prokaryotic cell strain. Int J Syst Evol Microbiol, 2004 May, 54(Pt 3), 951 - 4 Propionispora hippei sp . nov., a novel Gram-negative, spore-forming anaerobe that produces propionic acid; Abou-Zeid DM et al.; A Gram-negative, spore-forming anaerobe, KS(T), was isolated from an enrichment culture that was set up for anaerobic degradation of the aliphatic polyester poly(propylene adipate) . The strain had the cellular organization of Sporomusa, vibrio-shaped cells and terminal round spores, and fermented sugars and sugar alcohols to propionic and acetic acid . Based on the morphological and physiological features as well as on a 16S rRNA gene similarity of 98 %, it was grouped with Propionispora vibrioides . A relatively low DNA-DNA hybridization value with the type strain of this species (47 %), and differences in substrate utilization and spore morphology, suggested that the strain should be classified in a separate species, Propionispora hippei sp . nov., with KS(T) as the type strain (=DSM 15287(T)=ATCC BAA-665(T)). Int J Syst Evol Microbiol, 2004 May, 54(Pt 3), 843 - 6 Vibrio gallicus sp . nov., isolated from the gut of the French abalone Haliotis tuberculata; Sawabe T et al.; Five alginolytic, facultatively anaerobic, non-motile bacteria were isolated from the gut of the abalone Haliotis tuberculata . Phylogenetic analyses based on 16S rDNA data indicated that these strains are related to Vibrio wodanis, Vibrio salmonicida, Vibrio logei and Vibrio fischeri (but with <97 % 16S rRNA gene sequence similarity) . DNA-DNA hybridization and fluorescence amplified fragment length polymorphism fingerprinting demonstrated that the five strains constituted a single species that was different from all currently known vibrios . The name Vibrio gallicus sp . nov . (type strain, CIP 107863(T)=LMG 21878(T)=HT2-1(T); DNA G+C content, 43.6-44.3 mol%) is proposed for this novel taxon . Several phenotypic features were disclosed that discriminated V . gallicus from other Vibrio species: V . gallicus can be differentiated from Vibrio halioticoli on the basis of four traits (beta-galactosidase test and assimilation of three carbon compounds) and from Vibrio superstes by 16 traits. Biosens Bioelectron, 2004 Jul 15, 19(12), 1711 - 22 On line biomonitors used as a tool for toxicity reduction evaluation of in situ groundwater remediation techniques; Kuster E et al.; Success of groundwater remediation is typically controlled via snapshot analysis of selected chemical substances or physical parameters . Biological parameters, i.e . ecotoxicological assays, are rarely employed . Hence the aim of the study was to develop a bioassay tool, which allows an on line monitoring of contaminated groundwater, as well as a toxicity reduction evaluation (TRE) of different remediation techniques in parallel and may furthermore be used as an additional tool for process control to supervise remediation techniques in a real time mode . Parallel testing of groundwater remediation techniques was accomplished for short and long time periods, by using the energy dependent luminescence of the bacterium Vibrio fischeri as biological monitoring parameter . One data point every hour for each remediation technique was generated by an automated biomonitor . The bacteria proved to be highly sensitive to the contaminated groundwater and the biomonitor showed a long standing time despite the highly corrosive groundwater present in Bitterfeld, Germany . The bacterial biomonitor is demonstrated to be a valuable tool for remediation success evaluation . Dose response relationships were generated for the six quantitatively dominant groundwater contaminants (2-chlortoluene, 1,2- and 1,4-dichlorobenzene, monochlorobenzene, ethylenbenzene and benzene) . The concentrations of individual volatile organic chemicals (VOCs) could not explain the observed effects in the bacteria . An expected mixture toxicity was calculated for the six components using the concept of concentration addition . The calculated EC(50) for the mixture was still one order of magnitude lower than the observed EC(50) of the actual groundwater . The results pointed out that chemical analysis of the six most quantitative substances alone was not able to explain the effects observed with the bacteria . Thus chemical analysis alone may not be an adequate tool for remediation success evaluation in terms of toxicity reduction. Microbiology, 2004 May, 150(Pt 5), 1283 - 90 Influences of temperature, salinity and starvation on the motility and chemotactic response of Vibrio anguillarum; Larsen MH et al.; The role of growth factors for the motility and chemotaxis of the fish pathogen Vibrio anguillarum was determined . Cells of V . anguillarum were chemotactic to serine in the temperature range 5-25 degrees C and in 0.8-2.7 % NaCl . The chemotactic response was significantly higher at 25 degrees C than at 5 or 15 degrees C . Growth in medium with 1.5 % NaCl gave a higher response than growth with 3 % NaCl; when the salinity of the chemotaxis buffer was raised, the chemotactic response was reduced . The role of starvation was also studied; V . anguillarum showed a high chemotactic response after starvation for 2 and 8 days . Motility and chemotaxis are important virulence factors for this bacterium . Not only was the ability to perform chemotactic motility maintained after starvation, but also it was shown that starvation does not interfere with the ability of the organism to cause infection in rainbow trout after a bath challenge . The swimming speed was reduced at lower temperatures . Within the range of salinity and starvation studied, the motile cells swam with the same velocity, indicating that V . anguillarum under all the examined conditions has a functional flagellum and rotates it with constant speed . Phenamil, a specific inhibitor of Na(+)-driven flagella, reduced the motility of both starved and non-starved cells of V . anguillarum indicating that, in both cases, a Na(+) motive force drives the flagellum. Pediatr Infect Dis J, 2004 May, 23(5), 475 - 6 Otitis media associated with Vibrio alginolyticus in a child with pressure-equalizing tubes; Feingold MH et al.; Vibrio alginolyticus is an unusual cause of otitis media . Infection usually occurs in the presence of a chronically perforated eardrum or patent pressure-equalizing tube . Infection with V . alginolyticus can occur after even mild, brief exposure to seawater, and the interval between exposure to seawater and onset of clinical infection can be prolonged. Fish Shellfish Immunol, 2004 Feb, 16(2), 151 - 61 The immune response of the white shrimp Litopenaeus vannamei and its susceptibility to Vibrio infection in relation with the moult cycle; Liu CH et al.; The white shrimp Litopenaeus vannamei (8.0-14.4 g) was examined for haemocyte count, phenoloxidase activity, respiratory burst (release of superoxide anion), phagocytic activity, and clearance efficiency to the pathogen Vibrio alginolyticus in relation with moult cycle (postmoult, A, B; intermoult, C; premoult, D(0)/D(1)D(2)/D(3)) . Granular cells were the highest at C and D(0)/D(1)stage, and the lowest at A stage . Hyaline cells and THC (total haemocyte count) were higher at C stage, but lower at postmoult stages . Phenoloxidase activity was the highest at C stage, and the lowest at A stage . Respiratory burst was lower at A stage . Phagocytic activity of shrimps against V . alginolyticus decreased significantly at postmoult and premoult stages . Additionally, the clearance efficiency of shrimps to V . alginolyticus was significantly lower for shrimps at A stage than those at C stage . In another experiment, L . vannamei at different moult stages were injected with tryptic soy broth (TSB)-grown V . alginolyticus (1x10(5)cfu shrimp(-1)) and then held in 34% seawater . After 10 h, the mortality of V . alginolyticus-injected shrimps was significantly higher for shrimps at postmoult stage than those at intermoult stage . Over 48-120 h, the mortality of V . alginolyticus-injected shrimps was 50.0%, 33.3% and 40.0% at postmoult, intermoult and premoult stage, respectively . It is concluded that L . vannamei showed a decrease in resistance at A stage through a reduction of its haemocyte count, phenoloxidase activity, respiratory burst, phagocytic activity and clearance efficiency against V . alginolyticus. Fish Shellfish Immunol, 2004 Feb, 16(2), 93 - 105 Efficacy of a bivalent vaccine against eel diseases caused by Vibrio vulnificus after its administration by four different routes; Esteve-Gassent MD et al.; Vulnivaccine, a vaccine against vibriosis caused by Vibrio vulnificus serovar E (formerly biotype 2), confers acceptable levels of protection to eels after its administration by prolonged immersion in three doses . Recently, a new pathogenic serovar, named serovar A, has been isolated from vaccinated eels in a Spanish freshwater eel farm . The main objective of this work was to design a bivalent vaccine, and to study its effectiveness against the two pathogenic serovars . With this aim, eels weighing around 20 g were immunised with the bivalent vaccine by oral and anal intubation, intraperitoneal injection (i.p.) and prolonged immersion . The overall results indicated that: (i) the new vaccine delivered by oral and anal intubation induced protection levels higher than 80%, to that achieved after i.p . vaccination; (ii) oral and anal vaccination induced a significant systemic and mucosal immune response; (iii) the protection after vaccination by whichever routes was related to antibody titres in plasma; (iv) mucosal and systemic compartments showed different kinetics of antibody production; (v) evidence for passive transfer of antibodies from plasma to gut mucus were found after i.p . and anal vaccination, and finally, (vi) vaccination did not enhance the production of lysozyme, in plasma or mucus . In conclusion, this new vaccine is effective in protecting eels against vibriosis caused by the two eel-pathogenic serovars of V . vulnificus, the oral delivery system is a promising way which may be used in intensive culture facilities during the whole growth period of eels. Fish Shellfish Immunol, 2004 Mar, 16(3), 407 - 14 Oral vaccination of African catfish with Vibrio anguillarum O2: effect on antigen uptake and immune response by absorption enhancers in lag time coated pellets; Vervarcke S et al.; The impact on antigen uptake and antibody response by the addition of absorption enhancers to Vibrio anguillarum O2 antigen was studied in oral vaccination trials of African catfish (Clarias gariepinus) . Oral vaccination was achieved by feeding lag time coated pellets . The lag time coat prevents premature release of the encapsulated vaccine in the tank, before ingestion of the pellets by the fish . To monitor the antigen uptake, a competitive ELISA was used . The antibody response was measured using an indirect ELISA . Feeding of bacterin-layered pellets without absorption enhancers resulted in a rather low antigen uptake and antibody levels . The addition of absorption enhancers such as sodium salicylate, sodium caprate and vitamin E TPGS increased the serum antigen levels and specific antibody levels in the systemic circulation . Skin mucus antibody levels were higher after oral vaccination compared to the IP and control group . The addition of absorption enhancers in the oral groups further increased the antibody levels obtained in the skin mucus. Fish Shellfish Immunol, 2004 Mar, 16(3), 321 - 34 Effect of ammonia on the immune response of white shrimp Litopenaeus vannamei and its susceptibility to Vibrio alginolyticus; Liu CH et al.; Growth of Vibrio alginolyticus was not affected by TSB medium containing ammonia-N concentration in the range of 0-20 mg l(-1) . White shrimp Litopenaeus vannamei (7-12 g in the intermolt stage) were challenged with V . alginolyticus, which had been incubated for 24 h in the TSB medium containing different concentrations of ammonia-N (0, 1, 5 . 10 and 20 mg l(-1)) . There was no significant difference in cumulative mortality for shrimp incubated in the TSB medium containing 0, 1, 5, 10 and 20 mg l(-1)ammonia-N after 120 h of challenge . The shrimps were challenged with V . alginolyticus previously incubated in the TSB medium for 24 h, then placed in water containing concentrations of ammonia-N at 0.01 mg l(-1)(control), 1.10, 5.24, 11.10 and 21.60 mg l(-1) . Mortality of shrimp in 5.24, 11.10 and 21.60 mg l(-1)was significantly higher than those in the control solution (0.01 mg l(-1)) after 48-168 h . Shrimps which had been exposed to control, 1.10, 5.24, 11.10 and 21.60 mg l(-1)ammonia-N for 7 days were examined for THC (total haemocyte count), granular cells, hyaline cells, phenoloxidase activity, release of superoxide anion, superoxide dismutase (SOD) activity, phagocytic activity and clearance efficiency to V . alginolyticus . No significant difference in THC, hyaline cells and granular cells were observed among shrimps at different ammonia-N concentrations . Phenoloxidase activity however, decreased when the shrimps were exposed to 5.24 mg l(-1)ammonia-N and greater after 7 days . The release of superoxide anion increased significantly, whereas SOD activity decreased significantly at 21.60 mg l(-1)ammonia-N . With shrimps exposed to 11.21 and 21.22 mg l(-1)ammonia-N for 7 days, phagocytic activity and clearance efficiency to V . alginolyticus significantly decreased . It is therefore suggested that ammonia in water caused a depression in the immune response and an increase in mortality of L . vannamei from the V . alginolyticus infection. Fish Shellfish Immunol, 2004 Mar, 16(3), 295 - 306 The immune response of Taiwan abalone Haliotis diversicolor supertexta and its susceptibility to Vibrio parahaemolyticus at different salinity levels; Cheng W et al.; Addition of NaCl at 2.5% to 3.5% to tryptic soy broth (TSB) significantly increased the growth of Vibrio parahaemolyticus . Taiwan abalone Haliotis diversicolor supertexta held in 30 per thousand seawater were injected with V . parahaemolyticus grown in TSB containing NaCl at 0.5, 1.5, 2.5, 3.5 and 4.5% at a dose of 1.6 x 10(5)colony-forming units (cfu) abalone(-1) . After 48 h, the cumulative mortality was significantly higher for the abalone challenged with V . parahaemolyticus grown in 2.5% than those grown in 0.5 and 1.5% NaCl . In other experiments, abalones held in 30 per thousand seawater were injected with TSB-grown V . parahaemolyticus (1.6 x 10(5)cfu abalone(-1)), and then transferred to 20, 25, 30 and 35 per thousand seawater . All abalones held in 20 per thousand were killed in 48 h . The mortality of V . parahaemolyticus-injected abalone held in 30 per thousand was significantly lower over 24-120 h . Abalone held in 30 per thousand seawater and then transferred to 20, 25, 30 and 35 per thousand were examined for THC (total haemocyte count), phenoloxidase activity, respiratory burst, phagocytic activity and clearance efficiency of V . parahemolyticus after 24 and 72 h . The THC increased directly related with salinity levels . Phenoloxidase activity, respiratory burst, phagocytic activity and clearance efficiency of V . parahaemolyticus decreased significantly for the abalone in 20, 25 and 35 per thousand . It is concluded that the abalone transferred from 30 per thousand to 20, 25 and 35 per thousand had reduced immune ability and decreased resistance against V . parahaemolyticus infection. J Antimicrob Chemother, 2004 Jun, 53(6), 947 - 51 Epub 2004 Apr 29. New aminoglycoside acetyltransferase gene, aac(3)-Id, in a class 1 integron from a multiresistant strain of Vibrio fluvialis isolated from an infant aged 6 months; Ahmed AM et al.; OBJECTIVES: To characterize the molecular basis of antibiotic resistance in a multidrug-resistant clinical isolate of Vibrio fluvialis H-08942 . PATIENT AND METHODS: V . fluvialis H-08942 was isolated from a hospitalized infant aged 6 months suffering from cholera-like diarrhoea in India in 2002 . The broth microdilution method was used to determine the MICs of a range of antibiotics for this strain . PCR, DNA sequencing, Southern hybridization, cloning and expression were used to characterize the molecular basis of antibiotic resistances . RESULTS: V . fluvialis H-08942 showed resistance to chloramphenicol, streptomycin, spectinomycin, co-trimoxazole, ampicillin, furazolidone, nalidixic acid and gentamicin . A class 1 integron that contains a novel aminoglycoside acetyltransferase gene, aac(3)-Id, and aminoglycoside adenyltransferase gene, aadA7, was characterized . The aac(3)-Id gene product was found to share 50%, 45% and 44% identity to AAC(3)-Ic, AAC(3)-Ia, and AAC(3)-Ib, respectively . Both aac(3)-Id and aadA7 genes were cloned and expressed in Escherichia coli . Phylogenetic analysis suggested that the aac(3)-Id represents a fourth evolutionary lineage in the aminoglycoside acetyltransferase genes . Southern hybridization showed that this integron is located in the chromosome . CONCLUSIONS: In this study we identified a new type of aminoglycoside acetyltransferase gene, aac(3)-Id . In addition, this is the first report of identification of antibiotic resistance genes and a class 1 integron in V . fluvialis. Bioessays, 2004 May, 26(5), 463 - 8 Phylogeny of gamma-proteobacteria: resolution of one branch of the universal tree? Brown JR, Volker C. The reconstruction of bacterial evolutionary relationships has proven to be a daunting task because variable mutation rates and horizontal gene transfer (HGT) among species can cause grave incongruities between phylogenetic trees based on single genes . Recently, a highly robust phylogenetic tree was constructed for 13 gamma-proteobacteria using the combined alignments of 205 conserved orthologous proteins.1 Only two proteins had incongruent tree topologies, which were attributed to HGT between Pseudomonas species and Vibrio cholerae or enterics . While the evolutionary relationships among these species appears to be resolved, further analysis suggests that HGT events with other bacterial partners likely occurred; this alters the implicit assumption of gamma-proteobacteria monophyly . Thus, any thorough reconstruction of bacterial evolution must not only choose a suitable set of molecular markers but also strive to reduce potential bias in the selection of species . Int J Food Microbiol, 2004 Apr 15, 92(2), 207 - 15 Susceptibility of Vibrio parahaemolyticus to various environmental stresses after cold shock treatment; Lin C et al.; Vibrio parahaemolyticus was subjected to cold shock treatment at 20 or 15 degrees C for 2 or 4 h . The effect of cold shock on the survival of V . parahaemolyticus subjected to subsequent low temperature (5 and -18 degrees C) and other adverse conditions (47 degrees C, 6 ppm crystal violet, 1000 ppm H(2)O(2), 25 mM acetic acid and 25 mM lactic acid) was investigated . Regardless of the cold shock treatment, survival of V . parahaemolyticus increased when stored at 5 or -18 degrees C, while no increase in survival was noted for cells cold shocked in the presence of chloramphenicol . Cold shock treatment under the conditions tested, in general, enabled V . parahaemolyticus cells to survive better following subsequent challenge by crystal violet, while the cold-shocked organism was more susceptible to high temperature (47 degrees C), H(2)O(2) and organic acids (lactic and acetic acid) than the non-shocked cells . Furthermore, the temperature and time of the cold shock treatment affected the cold shock response of V . parahaemolyticus. FEMS Microbiol Lett, 2004 May 1, 234(1), 163 - 7 Significance of Na+ in the fish pathogen, Vibrio anguillarum, under energy depleted condition; Fujiwara-Nagata E et al.; Vibrio anguillarum kills various kinds of fish over salinities ranging from seawater to freshwater . In this study, we investigated the role of Na(+) in V . anguillarum, especially under energy-depleted conditions such as in natural seawater . V . angustum S14, which is a typical marine vibrio, was used for comparison . V . anguillarum only required Na(+) for starvation-survival, but in contrast, V . angustum S14 always required Na(+) for both growth and starvation-survival . In marine vibrios, Na(+) is used in the Na(+)-dependent respiratory chain that produces the sodium motive force (SMF) across the cell membrane . It has been considered that marine vibrios always need a SMF produced by Na(+), however in the case of V . anguillarum, the SMF is not required for growth, but becomes more important for starvation-survival. Dis Aquat Organ, 2004 Mar 10, 58(2-3), 223 - 30 Pathogenicity testing of shellfish hatchery bacterial isolates on Pacific oyster Crassostrea gigas larvae; Estes RM et al.; Bacterial diseases are a major cause of larval mortality in shellfish hatcheries . Even with proper sanitation measures, bacterial pathogens cannot be eliminated in all cases . The pathogenicity of bacteria isolated from Pacific Northwest shellfish hatcheries to Pacific oyster Crassostrea gigas larvae was investigated . We found 3 highly pathogenic strains and 1 mildly pathogenic strain among 33 isolates tested . These strains appear to be members of the genus Vibrio . Although there have been many studies of bivalve bacterial pathogens, a standard method to assess bacterial pathogenicity in bivalve larvae is needed . Thus, we developed 2 methods using either 15 ml conical tubes or tissue culture plates that were employed for rapidly screening bacterial strains for pathogenicity to Pacific oyster larvae . The tissue culture plates worked well for screening both mildly pathogenic strains and LD50 (lethal dose) assays . This method allowed for non-intrusive and non-destructive observation of the oyster larvae with a dissecting microscope . The LD50 for the 3 highly pathogenic strains ranged between 1.6 and 3.6 x 10(4) colony forming units (CFU) ml(-1) after 24 h and between 3.2 x 102 and 1.9 x 10(3) CFU ml(-1) after 48 h. Dis Aquat Organ, 2004 Mar 10, 58(2-3), 143 - 50 Phylogenetic study and identification of Vibrio splendidus-related strains based on gyrB gene sequences; Le Roux F et al.; Different strains related to Vibrio splendidus have been associated with infection of aquatic animals . An epidemiological study of V . splendidus strains associated with Crassostrea gigas mortalities demonstrated genetic diversity within this group and suggested its polyphyletic nature . Recently 4 species, V . lentus, V . chagasii, V . pomeroyi and V . kanaloae, phenotypically related to V . splendidus, have been described, although biochemical methods do not clearly discriminate species within this group . Here, we propose a polyphasic approach to investigate their taxonomic relationships . Phylogenetic analysis of V . splendidus-related strains was carried out using the nucleotide sequences of 16S ribosomal DNA (16S rDNA) and gyrase B subunit (gyrB) genes . Species delineation based on 16S rDNA-sequencing is limited because of divergence between cistrons, roughly equivalent to divergence between strains . Despite a high level of sequence similarity, strains were separated into 2 clades . In the phylogenetic tree constructed on the basis of gyrB gene sequences, strains were separated into 5 independent clusters containing V . splendidus, V . lentus, V . chagasii-type strains and a putative new genomic species . This phylogenetic grouping was almost congruent with that based on DNA-DNA hybridisation analysis . V . pomeroyi, V . kanaloae and V . tasmaniensis-type strains clustered together in a fifth clade . The gyrB gene-sequencing approach is discussed as an alternative for investigating the taxonomy of Vibrio species. Microbiol Immunol, 2004, 48(4), 319 - 27 Emergence and serovar transition of Vibrio parahaemolyticus pandemic strains isolated during a diarrhea outbreak in Vietnam between 1997 and 1999; Chowdhury A et al.; We characterized 523 Vibrio parahaemolyticus strains isolated during a survey of diarrhea patients in Khanh Hoa province, Vietnam between 1997 and 1999 . Forty-nine percent of the strains were judged to belong to the pandemic strains that emerged around 1996 and spread to many countries . These strains were positive in the GS-PCR assay and carried the tdh gene . The ORF8 of the f237 phage genome, a possible marker of the pandemic clone, was absent in 10% of these strains . Eleven O: K serovars were detected among the pandemic strains and the strains representing all 11 serovars of pandemic strains were shown to be closely related regardless of the ORF8 genotype using arbitrarily primed PCR and pulsed field gel electrophoresis analyses . It was clear that a transition of major serovars occurred among the pandemic strains represented by the emergence of O3: K6 in 1997, O4: K68 in 1998, and O1: K25 in 1998 and 1999. Microbiol Immunol, 2004, 48(4), 313 - 8 Cytotoxicity and enterotoxicity of the thermostable direct hemolysin-deletion mutants of Vibrio parahaemolyticus; Park KS et al.; The thermostable direct hemolysin (TDH) has been proposed to be a major virulence factor of Vibrio parahaemolyticus . We have recently completed the genome sequence of a TDH-producing V . parahaemolyticus strain, RIMD2210633 . In this study, we constructed tdh-deletion mutants from the sequenced strain by homologous recombination and analyzed their phenotypes . Although the deletion of both copies of tdh completely abolished the hemolytic activity of the wild-type strain, the deletion did not affect the cytotoxicity to HeLa cells . Enterotoxicity, assayed by the rabbit ileal loop test, was lowered by tdh deletion, but the mutant still showed partial fluid accumulation in rabbit intestine . These results indicate that the cytotoxicity and enterotoxicity of TDH-producing V . parahaemolyticus are not explained by TDH alone, and suggest that an unknown virulence factor(s) could be involved in these pathogenic activities. Microbiol Immunol, 2004, 48(4), 243 - 50 A major Li(+) extrusion system NhaB of Pseudomonas aeruginosa : comparison with the major Na(+) extrusion system NhaP; Kuroda T et al.; A gene encoding a Li(+) extrusion system was cloned from the chromosomal DNA of Pseudomonas aeruginosa and expressed in Escherichia coli cells . The gene enabled growth of E . coli KNabc cells, which were unable to grow in the presence of 10 mM LiCl or 0.1 M NaCl because of the lack of major Na(+) (Li(+))/H(+) antiporters . We detected Li(+)/H(+) and Na(+)/H(+) antiport activities in membrane vesicles prepared from E . coli KNabc cells that harbored a plasmid carrying the cloned gene . Activity of this antiporter was pH-dependent with an optimal pH activity between pH 7.5 and 8.5 . These properties indicate that this antiporter is different from NhaP, an Na(+)/H(+) antiporter from P . aeruginosa that we reported previously, and that is rather specific to Na(+) but it cannot extrude Li(+) effectively . The gene was sequenced and an open reading frame (ORF) was identified . The amino acid sequence deduced from the ORF showed homology (about 60% identity and 90% similarity) with that of the NhaB Na(+)/H(+) antiporters of E . coli and Vibrio parahaemolyticus . Thus, we designated the antiporter as NhaB of P . aeruginosa . E . coli KNabc carrying the nhaB gene from P . aeruginosa was able to grow in the presence of 10 to 50 mM LiCl, although KNabc carrying nhaP was unable to grow in these conditions . The antiport activity of NhaB from P . aeruginosa was produced in E . coli and showed apparent Km values for Li(+) and Na(+) of 2.0 mM and 1.3 mM, respectively . The antiport activity was inhibited by amiloride with a Ki value for Li(+) and Na(+) of 0.03 mM and 0.04 mM, respectively. Microbiol Immunol, 2004, 48(4), 229 - 35 Virulence properties of rough and smooth strains of Vibrio cholerae O1; Islam MS et al.; A comparative study was carried out to see the differences in pathogenicity of rough and smooth strains . A total of 10 strains including 5 each of rough and smooth strains of Vibrio cholerae O1 were tested and found positive for toxin production by enzyme-linked immunosorbent assay (ELISA) in Richardson's and AKI media . All the smooth and rough strains, except one, showed a titre of 1: 10 and 1: 100 in Richardson's and AKI media, respectively . Both types of strains produced enterotoxin in rabbit ileal loop (RIL) . The differences in multiplication abilities of smooth and rough strains in RIL were statistically significant (P <0.05) . However, these differences in multiplying abilities did not influence the adherence potential or enterotoxin production as there was no significant difference (P >0.05) between these properties . This study demonstrated that the rough strains are equally pathogenic and as important as smooth strains. Arch Environ Contam Toxicol, 2004 Feb, 46(2), 176 - 82 Effect of pH on arsenate and arsenite toxicity to luminescent bacteria (Vibrio fischeri); Fulladosa E et al.; Arsenic is an abundant metalloid and a dangerous pollutant when in solution under the arsenate or arsenite forms-As(V) and As(III), respectively . Since its biological effects are expected to depend on the oxidation state and on speciation, effect of pH on either As(V) or As(III) speciation and resulting toxicity was investigated using the Microtox bioassay based on change in light emission by the luminescent bacteria Vibrio fischeri . Within a 5.0-8.0 pH range, EC50 values for As(V) were found to decrease as pH became basic, reflecting an increase in toxicity; whereas in the case of As(III), EC50 values were almost unchanged within a 6.0-8.0 pH range and lowered only at pH 9.0 . HAsO42- and H2AsO3-were found to be the most toxic species . A statistical approach based on testing the null hypothesis of additive toxicity revealed an antagonistic effect between the arsenate chemical species . At low concentrations, As(V) was regularly found to be more toxic than As(III), independent of the pH value . Conversely, at high concentrations, the toxicity of both As(III) and As(V) was found to chiefly depend on pH, as a consequence of the strong influence of this parameter on the chemical speciation. Kansenshogaku Zasshi, 2004 Feb, 78(2), 83 - 9 {A basic study of Vibrio vulnificus infection: serotyping and drug sensitivity test of environment-derived strains and human clinical isolates}; Oonaka K et al.; In attempt to elucidate the route and source of Vibrio vulnificus infection . serotyping and drug sensitivity tests of environment-derived strains and human clinical isolates were performed . 1) Serotyping of isolates from the two types of source were determined . Of environment-derived strains, 72.5% were classified into 18 types, and O7 was the most frequent type, accounting for 73.1%, and the second frequent type was O4, accounting for 6.1% . Of human clinical isolates, 87.1% were classified into eight types, and O4 was the most frequent, accounting for 73.5%, and O7 was the secondly most frequent, accounting for 12.9% . 2) Serotypes were investigated by regions . In eastern Japan, 69.2% were classified into 18 types, and O7 and O4 accounting for 44.6% and 5.7%, respectively . In western Japan, 64.8% were classified into eight types, and O7 was the most frequent, accounting for 20.4%, and secondly frequent type was O4, accounting for 11.1% . 3) Regarding the relationship between biotypes and serotypes, environment-derived biotype-I strains were widely distributed in the serotypes, but most biotype-I human clinical isolates were distributed in serotypes O1-O7, showing a difference between the two types of sources . However, many biotype-II strains from the two types of sources included in the serotype O7 group . 4) Drug sensitivity was compared based on MIC90 between strains from the two types of sources . Environment-derived strains were sensitive to ABPC, PIPC, CPZ, CTX, LMOX, MEPM, GM, EM, TC, DOXY, MINO, CP, NA and CPFX, but some strains were resistant to CER, CET, CTX, CMZ, KM and LCM . Human clinical isolates were sensitive to EM, TC, DOXY, MINO, CP, NA and CPFX, but some strains were resistant to ABPC, PIPC, CER, CET, CPZ, CTX, CMZ, LMOX, MEPM, KM, GM, AMK and LCM. Environ Toxicol, 2004 Jun, 19(3), 191 - 7 Anatoxin-a toxin in the cyanobacterium Planktothrix rubescens from a fishing pond in northern Italy; Viaggiu E et al.; A heavy algal bloom occurring in a fishing pond in northern Italy full of Salmo trutta was examined for algae taxonomy and toxic production . The dominant algal species (98%) was identified as the cyanobacterium Planktothrix rubescens (D.C . ex GOMONT) Komarek Anagnostidis, based on morphological examination, and it was revealed to be toxic in mouse and Vibrio fischeri bioassays . The toxin was identified as anatoxin-a using high-performance liquid chromatography and confirmed using liquid chromatography-mass spectrometry (LC-MS) . The mouse bioassay gave signs of poisoning, as previously reported for anatoxin-a . The LC-MS confirmed the presence of an anatoxin-a peak at m/z 166 (M+H+) . The content of toxin in the field population was estimated at 12.13 microg/g of fresh cells . The bloom was sustained by the very high N/P ratio in the water . This is the first report in Italy of an anatoxin-a-producing Planktothrix rubescens population . Clin Infect Dis, 2004 Apr 15, 38 Suppl 3, S203 - 11 Survey of physician diagnostic practices for patients with acute diarrhea: clinical and public health implications; Hennessy TW et al.; To understand physician practices regarding the diagnosis of acute diarrheal diseases, we conducted a survey, in 1996, of 2839 physicians in Connecticut, Georgia, Minnesota, Oregon, and California . Bacterial stool culture was requested for samples from the last patient seen for acute diarrhea by 784 (44%; 95% confidence interval, 42%-46%) of 1783 physicians . Physicians were more likely to request a culture for persons with acquired immune deficiency syndrome, bloody stools, travel to a developing country, diarrhea for >3 days, intravenous rehydration, or fever . Substantial geographic and specialty differences in culture-request practices were observed . Twenty-eight percent of physicians did not know whether stool culture included testing for Escherichia coli O157:H7; 40% did not know whether Yersinia or Vibrio species were included . These variabilities suggest a need for clinical diagnostic guidelines for diarrhea . Many physicians could benefit from education to improve their knowledge about tests included in routine stool examinations. Cell Mol Life Sci, 2004 Apr, 61(7-8), 961 - 72 Involvement of penaeidins in defense reactions of the shrimp Litopenaeus stylirostris to a pathogenic vibrio; Munoz M et al.; The present study reports for the first time the involvement of an antimicrobial peptide in the defense reactions of a shrimp infected by a pathogenic Vibrio, Vibrio penaeicida . New members of the penaeidin family were characterized in the shrimp Litopenaeus stylirostris by RT-PCR and RACE-PCR from hemocyte total RNAs, and by mass spectrometry detection and immunolocalization of mature peptides in shrimp hemocytes . In infected shrimps, bacteria and penaeidin distribution colocalized in the gills and the lymphoid organ that represented the main infected sites . Moreover, the shrimp immune response to infection involved massive hemocyte recruitment to infection sites where released penaeidin may participate in the isolation and elimination of the bacteria, We show that the ability of the shrimps to circumvent shrimp infections is closely related to a recovery phase based on the hematopoietic process. Toxicol Appl Pharmacol, 2004 May 1, 196(3), 319 - 26 Lysophospholipase inhibition by organophosphorus toxicants; Quistad GB et al.; Lysophospholipases (LysoPLAs) are a large family of enzymes for removing lysophospholipids from cell membranes . Potent inhibitors are needed to define the importance of LysoPLAs as targets for toxicants and potential therapeutics . This study considers organophosphorus (OP) inhibitors with emphasis on mouse brain total LysoPLA activity relative to the mipafox-sensitive neuropathy target esterase (NTE)-LysoPLA recently established as 17% of the total activity and important in the action of OP delayed toxicants . The most potent inhibitors of total LysoPLA in mouse brain are isopropyl dodecylphosphonofluoridate (also for LysoPLA of Vibrio bacteria), ethyl octylphosphonofluoridate (EOPF), and two alkyl-benzodioxaphosphorin 2-oxides (BDPOs){(S)-octyl and dodecyl} (IC50 2-8 nM) . OP inhibitors acting in vitro and in vivo differentiate a more sensitive portion but not a distinct NTE-LysoPLA compared with total LysoPLA activity . For 10 active inhibitors, NTE-LysoPLA is 17-fold more sensitive than total LysoPLA, but structure-activity comparisons give a good correlation (r(2) = 0.94) of IC50 values, suggesting active site structural similarity or identity . In mice 4 h after intraperitoneal treatment with discriminating doses, EOPF, tribufos (a plant defoliant), and dodecanesulfonyl fluoride inhibit 41-57% of the total brain LysoPLA and 85-99% of the NTE-LysoPLA activity . Total LysoPLA as well as NTE-LysoPLA is decreased in activity in Nte(+/-)-haploinsufficient mice compared to their Nte(+/+) littermates . The lysolecithin level of spinal cord but not brain is elevated significantly following EOPF treatment (3 mg/kg), thereby focusing attention on localized rather than general alterations in lysophospholipid metabolism in OP-induced hyperactivity and toxicity. Biochim Biophys Acta, 2004 Apr 16, 1678(1), 7 - 13 Cloning and characterization of the lipase and lipase activator protein from Vibrio vulnificus CKM-1; Su JH et al.; The gene (lipA) encoding the extracellular lipase and its downstream gene (lipB) from Vibrio vulnificus CKM-1 were cloned and sequenced . Nucleotide sequence analysis and alignments of amino acid sequences suggest that Lip Ais a member of bacterial lipase family I.1 and that LipB is a lipase activator of LipA . The active LipA was produced in recombinant Escherichia coli cells only in the presence of the lipB . In the hydrolysis of p-nitrophenyl esters and triacylglycerols, using the reactivated LipA, the optimum chain lengths for the acyl moiety on the substrate were C(14) for ester hydrolysis and C(10) to C(12) for triacylglycerol hydrolysis. Environ Pollut, 1992, 76(3), 245 - 9 Effects of temperature and pH on conjugal transfer of zinc-resistant plasmids residing in Gram-negative bacteria isolated from industrial effluents; Hasnain S et al.; Two zinc (Zn)-resistant strains, AnZn-1 and AnZn-2, which were resistant to ZnSO(4) up to 12.5 mg ml(-1) were isolated from industrial effluents . Both were Gram-negative with motile cells . They exhibited tolerance to Ba(2+), Ni(+), Co(2+) Mn(2+), Cu(2+), Fe(2+), Ni(2+), Cd(2+), kanamycin, chloramphenicol, ampillicin and tetracycline, but were sensitive to Hg(2+) and streptomycin . For AnZn-1 and AnZn-2, the optimum pH for growth was 7 . Both were facultative anaerobes and had cytochrome oxidase and urease enzymes, while catalase was present only in AnZn-2 . Both strains had the ability to hydrolyse gelatin, reduce nitrate, and yield acid from arabinose and rhamnose . The two strains shared maximum characters with Vibrionaceae . Each strain carries a single Zn-resistant conjugative plasmid . The plasmid residing in AnZn-1 (pSH1211) displayed a lower level of resistance than the plasmid of AnZn-2 (pSH1212) . Both required a minimum of 24 h for mating and showed highest transfer frequency at 25 degrees C . pSH1211 preferred pH 7 and pSH1212 pH 8.5 for their transfer . Both plasmids, when allowed to mate with Escherichia coli at 25 degrees C, alkaline pH values of 8-8.5 (pSH1211) of pH 7.5 (pSH1212), showed increased transfer frequency. Blood Coagul Fibrinolysis, 2004 Mar, 15(2), 169 - 78 A secreted metallo protease from Aeromonas hydrophila exhibits prothrombin activator activity; Keller T et al.; Detection, purification, and partial characterization of a protease from Aeromonas hydrophila capable of cleaving prothrombin into active thrombin is described . The protease has been characterized with respect to enzymatic characteristics such as optimum reaction conditions for prothrombin activation, usage of additional substrates, as well as sensitivity against inhibitors . The protease activity can reversibly be inhibited by Me2+ chelating agents like ethylenediamine tetraacetic acid . The enzyme exhibits a pI value of 4.4 and can withstand temperatures up to 55 degrees C without loss of activity . With respect to prothrombin the enzyme exhibits a K(M) value of 1.47 micromol/l and a vmax value of 1.66 mol/min per mol enzyme . Amino terminal sequence analysis as well as mass spectrometry of fragments obtained by trypsin digest showed identity to a recently described elastase type protease from the same organism and homology to known proteases from other procaryotes (e.g . Aeromonas caviae, Vibrio proteolytica, Pseudomonas aeruginosa). J Bacteriol, 2004 May, 186(9), 2906 - 8 Correlation between osmolarity and luminescence of symbiotic Vibrio fischeri strain ES114; Stabb EV et al.; Vibrio fischeri isolates from Euprymna scolopes are dim in culture but bright in the host . We found the luminescence of V . fischeri to be correlated with external osmolarity both in culture and in this symbiosis . Luminescence enhancement by osmolarity was independent of the lux promoter and unaffected by autoinducers or the level of lux expression, but the addition of an aldehyde substrate for luciferase raised the luminescence of cells grown at high and low osmolarities to the same high level . V . fischeri culture media have lower osmolarities than are typical in seawater or in cephalopods, partially accounting for the bacterium's low light output in culture. J Bacteriol, 2004 May, 186(9), 2636 - 45 Formation of SXT tandem arrays and SXT-R391 hybrids; Burrus V et al.; SXT is an integrative and conjugative element (ICE) isolated from Vibrio cholerae . This approximately 100-kb ICE encodes resistance to multiple antibiotics and integrates site specifically into the chromosome . SXT excises from the chromosome to form a circular but nonreplicative extrachromosomal molecule that is required for its transfer . Here we found that a significant fraction of freshly isolated SXT exconjugants contained tandem SXT arrays . There was heterogeneity in the size of the SXT arrays detected in single exconjugant colonies . Some arrays consisted of more than five SXTs arranged in tandem . These extended arrays were unstable and did not persist during serial passages . The mechanism accounting for the generation of SXT arrays is unknown; however, array formation was not dependent upon recA and appeared to depend on conjugative transfer . While such arrays did not alter the transfer frequency of wild-type SXT, they partially complemented the transfer deficiency of a Deltaxis SXT mutant, which is ordinarily unable to generate the extrachromosomal intermediate required for SXT transfer . Exconjugants derived from donor strains that harbored tandem arrays of SXT and R391, an SXT-related element, contained functional hybrid elements that arose from recA-independent recombination between the two ICEs . Thus, arrays of SXT-related elements promote the creation of novel ICEs. J Bacteriol, 2004 May, 186(9), 2558 - 66 The bchU gene of Chlorobium tepidum encodes the c-20 methyltransferase in bacteriochlorophyll c biosynthesis; Maresca JA et al.; Bacteriochlorophylls (BChls) c and d, two of the major light-harvesting pigments in photosynthetic green sulfur bacteria, differ only by the presence of a methyl group at the C-20 methine bridge position in BChl c . A gene potentially encoding the C-20 methyltransferase, bchU, was identified by comparative analysis of the Chlorobium tepidum and Chloroflexus aurantiacus genome sequences . Homologs of this gene were amplified and sequenced from Chlorobium phaeobacteroides strain 1549, Chlorobium vibrioforme strain 8327d, and C . vibrioforme strain 8327c, which produce BChls e, d, and c, respectively . A single nucleotide insertion in the bchU gene of C . vibrioforme strain 8327d was found to cause a premature, in-frame stop codon and thus the formation of a truncated, nonfunctional gene product . The spontaneous mutant of this strain that produces BChl c (strain 8327c) has a second frameshift mutation that restores the correct reading frame in bchU . The bchU gene was inactivated in C . tepidum, a BChl c-producing species, and the resulting mutant produced only BChl d . Growth rate measurements showed that BChl c- and d-producing strains of the same organism (C . tepidum or C . vibrioforme) have similar growth rates at high and intermediate light intensities but that strains producing BChl c grow faster than those with BChl d at low light intensities . Thus, the bchU gene encodes the C-20 methyltransferase for BChl c biosynthesis in Chlorobium species, and methylation at the C-20 position to produce BChl c rather than BChl d confers a significant competitive advantage to green sulfur bacteria living at limiting red and near-infrared light intensities. Chem Res Toxicol, 2004 Apr, 17(4), 545 - 54 Assessment and modeling of the toxicity of organic chemicals to Chlorella vulgaris: development of a novel database; Cronin MT et al.; This study reports a database of toxicity values for 91 compounds assessed in a novel, rapid, and economical 15 min algal toxicity test . The toxicity data were measured using the unicellular green alga Chlorella vulgaris in an assay that determined the disappearance of fluorescein diacetate . The chemicals tested covered a wide range of physicochemical properties and mechanisms of action . Quantitative activity-activity relationships with the toxicity of the chemicals to other species (Tetrahymena pyriformis, Vibrio fischeri, and Pimephales promelas) showed strong relationships, although some differences resulting from different protocols were established . Quantitative structure-activity relationships (QSARs) were determined using linear {multiple linear regression (MLR)} and nonlinear {k-nearest neighbors (KNN)} methods . Three descriptors, accounting for hydrophobicity, electrophilicity, and a function of molecular size corrected for the presence of heteroatoms, were found to be important to model toxicity . The predictivity of MLR was compared to KNN using leave-one-out cross-validation and the simulation of an external test set . MLR demonstrated greater stability in validation . The results of this study showed that method selection in QSAR is task-dependent and it is inappropriate to resort to more complicated but less transparent methods, unless there are clear indications (e.g., inability of MLR to deal with the data set) for the need of such methods. Bacteriol Virusol Parazitol Epidemiol, 2002 Jul-Dec, 47(3-4), 119 - 24 {Factors influencing the capacity of cellular substrate adherence of vibrio cholerae O1 and non O1 strains}; Israil AM et al.; Bacterial adherence to eukariotic cells represents an important step of tissue colonization and is mediated by specific molecules called adhesins . Bacterial adherence to cellular substrate is a very complex process consisting in specific interactions between the surface of host cell and bacterial cell surface respectively . Adherence to cellular substrate confers selective advantages to bacterial cells, as: rapid growth rate by shorter lag period and protection against antibodies and lysozime . Adherence and colonization of small bowel represent the early steps of cholera infection (1, 2) . The purposes of this study were to characterize the adherence ability of 46 Vibrio cholerae O1 and non O1 strains with different sources of isolation (acute diarrhea, water sources) to HEp-2 cell; to determine the influence of different factors (culture media, bacterial culture growth phase, proteolytic enzymes, carbohydrates and polyvalent agglutinant anti V . cholerae O1 serum) on the bacterial adherence capacity . Adherence capacity was assayed using the qualitative Cravioto's method . The adherence ability was appreciated by semi quantitative ("+", "++" and "+++") and quantitative assays . The adherence pattern of the tested strains was predominantly a diffuse one . The agar medium proved to be the most appropriate for the early and maximal expression of adhesion molecules, by comparison with nutritive broth and alkaline peptone water . Manose in different concentrations (1% and 3%) inhibited the adherence ability, demonstrating the role of manose-sensitive haemagglutinating fimbriae (MSHA) in mediating the adherence of V . cholerae strains to cellular substrate . Trypsine has no notable effect on the adherence ability, suggesting that the major V . cholerae adhesion molecules are not essentially of protein nature, so that the afimbrial adhesins could also play an important role in bacterial adhesion to eukariotic cells . The agglutinant polyvalent anti-V . cholerae O1 serum had the most significant inhibitory effect on the adherence ability, which was completely abolished in the presence of sub-agglutinant dilutions of serum titer (1/60-1/120) and partially reduced at titers ranging from 1/240 to 1/920 . This inhibitory effect could be explained by bacterial agglutination, but also by the specific blocking of some surface structure implicated in the adherence process (i.e . lipopolysaccharides, as demonstrated by the inhibitory effect of sub-agglutinant serum titers) . The inhibitory effect of polyvalent anti-V . cholerae O1 serum was limited to O1, but was not evident for the non O1 serogroups, demonstrating that the serum antibodies are acting on serogroup specific antigenic fractions. J Mol Biol, 2004 Apr 30, 338(3), 585 - 96 The crystal structure of the periplasmic domain of the type II secretion system protein EpsM from Vibrio cholerae: the simplest version of the ferredoxin fold; Abendroth J et al.; The terminal branch of the general secretion pathway (Gsp or type II secretion system) is used by several pathogenic bacteria for the secretion of their virulence factors across the outer membrane . In these secretion systems, a complex of 12-15 Gsp proteins spans from the pore in the outer membrane via several associated signal or energy-transducing proteins in the inner membrane to a regulating ATPase in the cytosol . The human pathogen Vibrio cholerae uses such a system, called the Eps system, for the export of the cholera toxin and other virulence factors from its periplasm into the lumen of the gastrointestinal tract of the host . Here, we report the atomic structure of the periplasmic domain of the EpsM protein from V.cholerae, which is a part of the interface between the regulating part and the rest of the Eps system . The crystal structure was determined by Se-Met MAD phasing and the model was refined to 1.7A resolution . The monomer consists of two alphabetabeta-subdomains forming a sandwich of two alpha-helices and a four-stranded antiparallel beta-sheet . In the dimer, a deep cleft with a polar rim and a hydrophobic bottom made by conserved residues is located between the monomers . This cleft contains an extra electron density suggesting that this region might serve as a binding site of an unknown ligand or part of a protein partner . Unexpectedly, the fold of the periplasmic domain of EpsM is an undescribed circular permutation of the ferredoxin fold. J Virol Methods, 2004 Jun 15, 118(2), 95 - 100 The use of degenerate-primed random amplification of polymorphic DNA (DP-RAPD) for strain-typing and inferring the genetic similarity among closely related viruses; Comeau AM et al.; Often it is necessary to distinguish among strains of closely related viruses, as well as infer the genetic relatedness within large groups of viruses . Current methods for strain-typing viruses are time-consuming, require significant quantities of extracted DNA, and/or may require a priori genetic information . In this study we modified random amplification of polymorphic DNA (RAPD) by using a degenerate primer to produce unique and reproducible banding patterns from viral genomes . In the degenerate-primed RAPD analysis (DP-RAPD), a selection of algal virus and bacteriophage strains were profiled that encompassed an array of genome sizes and virus families . Closely related viruses (e.g . strains infecting Micromonas pusilla) generated similar, yet unique DP-RAPD patterns that could be readily distinguished from viruses within the same family (Phycodnaviridae) infecting a Chlorella-like alga . As well, marine vibriophage from the families Myoviridae, Siphoviridae, and Podoviridae showed high diversity and were distinct from coliphage and cyanophage . Contamination of host DNA, even at levels above those that would normally be encountered, did not interfere with the viral patterns . These findings describe a rapid, PCR-based tool for strain-typing viral isolates that allows inferences to be made on genetic relatedness within groups of closely related viruses. Water Sci Technol, 2004, 49(4), 273 - 7 Effect of advanced oxidation processes on the toxicity of municipal landfill leachates; Slomczynska B et al.; The aim of the present study was to assess the effect of advanced oxidation processes (AOPs) (oxidation ozone and peroxide/ozone) on the toxicity of leachates from municipal landfill for Warsaw, Poland, using a battery of tests . AOPs used to pre-treat leachates were carried out in laboratory conditions after their coagulation with the use of FeCl3 . The effects of the pre-treatment of leachates using the method of coagulation with FeCl3 depended on the concentration of organic compounds and with optimal conditions of the process ranged from 40 to 70% . Further pre-treatment of the leachates after coagulation, involving the use of oxidation with 03 and H2O2/O3, did not cause significant decrease of leachate toxicity . The data of this study demonstrated the usefulness of the battery of tests using Daphnia magna, Artemia franciscana, Scenedesmus quadricauda and Vibrio fischeri for the toxicity evaluation of raw and pre-treated leachates. Mikrobiologiia, 2004 Jan-Feb, 73(1), 80 - 8 {Ectothiorhodosinus mongolicum gen . nov., sp . nov.,--a new purple sulfur bacterium from soda lake in Mongolia}; Gorlenko VM et al.; A new purple sulfur bacterium (strain M9) was isolated from the steppe soda Lake Dzun Uldziin Nur (pH 9.4; mineralization, 3.3%) situated in southeastern Mongolia . Individual cells appear as vibrios 0.3-0.5 x 0.7-1 micron in size . The dividing cells often do not separate from each other, forming an almost closed ring . The internal photosynthetic membranes are represented by concentric lamellae lining the cell wall . Photosynthetic pigments are bacteriochlorophyll a and carotenoids of the spirilloxanthin series . The main carotenoid (> 96%) is spirilloxanthin . Two typical light-harvesting complexes (LH1 and LH2) are present in the membranes in a 1:1 ratio . The bacterium is an anaerobe and facultative photoorganoheterotroph . Photolithoautotrophic growth on sulfide is scarce . Thiosulfate is utilized as an electron donor only in the presence of organic matter . Globules of elemental sulfur are formed as an intermediary product of sulfide and thiosulfate oxidation and are deposited outside the cells . The end product of oxidation is sulfate . In the presence of sulfide and carbonates, acetate, lactate, malate, pyruvate, propionate, succinate, and fumarate are used as the additional sources of carbon in anoxygenic photosynthesis . Vitamin are not required . The bacterium is an alkaliphile the pH optimum is at 8.3-9.1, the pH range is 7.6-10.1 . The optimum NaCl concentration in the medium is 1 to 7%; the range is 0.5 to 0.9% . The optimum carbonate content in the medium is 2%; the range is 1 to 10% . The best growth occurs at 30-35 degrees C . The DNA G + C content is 57.5 mol% . According to the results of analysis of the 16S rRNA gene sequences, the new isolate M9 belongs to the phylogenetic cluster containing representatives of the family Ectothiorhodospiraceae within the class "Gammaproteobacteria." In this class, the new isolate forms a new branch, which occupies an intermediate position between the representatives of the genera Ectothiorhodospira and Thiorhodospira . Based on the phenotypic and genetic characteristics, the new purple sulfurbacterium was assigned to a new species of a new genus of the family Ectothiorhodospiraceae, Ectothiorhodosinus mongolicum gen . nov., sp . nov. Microbiology, 2004 Apr, 150(Pt 4), 911 - 20 Isolation of Vibrio alginolyticus sodium-driven flagellar motor complex composed of PomA and PomB solubilized by sucrose monocaprate; Yakushi T et al.; The polar flagella of Vibrio alginolyticus have sodium-driven motors, and four membrane proteins, PomA, PomB, MotX and MotY, are essential for torque generation of the motor . PomA and PomB are believed to form a sodium-conducting channel . This paper reports the purification of the motor complex by using sucrose monocaprate, a non-ionic detergent, to solubilize the complex . Plasmid pKJ301, which encodes intact PomA, and PomB tagged with a C-terminal hexahistidine that does not interfere with PomB function, was constructed . The membrane fraction of cells transformed with pKJ301 was solubilized with sucrose monocaprate, and the solubilized materials were applied to a Ni-NTA column . The imidazole eluate contained both PomA and PomB, which were further purified by anion-exchange chromatography . Gel-filtration chromatography was used to investigate the apparent molecular size of the complex; the PomA/PomB complex was eluted as approx . 900 kDa and PomB alone was eluted as approx . 260 kDa . These findings suggest that the motor complex may have a larger structure than previously assumed. Commun Dis Intell, 2004, 28(1), 6 - 68 Australia's notifiable diseases status, 2002: Annual report of the National Notifiable Diseases Surveillance System; Yohannes K et al.; There were 57 infectious diseases notifiable at the national level in Australia in 2002 . States and territories reported 100,278 cases of infectious diseases to the National Notifiable Diseases Surveillance System (NNDSS), a fall of 4 per cent compared to the number of notifications in 2001 . In 2002, the most frequently notified diseases were, sexually transmitted infections (31,929 reports, 32% of total notifications), gastrointestinal infections (26,708 reports, 27% of total notifications) and bloodborne infections (23,741, 24%) . There were 11,711 (12% of total) cases of vaccine preventable diseases, 3,052 (3% of total) cases of vectorborne diseases, 1,155 (1% of total) cases of zoonotic infections, two cases of quarantinable diseases (Vibrio cholerae O1) and 1,980 cases of other bacterial diseases, notified to NNDSS . Compared to 2001, notifications of sexually transmitted infections increased by 16 per cent and gastrointestinal infections by 2 per cent while bloodborne infections fell by 18 per cent . The number of notifications of chlamydial infection and Q fever were the highest since 1991 and 1995 respectively . By contrast, the number of notification for hepatitis A and measles were the lowest since 1991 . For other notifiable diseases, the number of notifications was within the range of the five years between 1997 and 2002 (range = five-year mean plus or minus two standard deviations) . This report also includes 2002 summary data on communicable diseases from other surveillance systems including the Laboratory Virology and Serology Reporting Scheme and sentinel general practitioner schemes. J Clin Microbiol, 2004 Apr, 42(4), 1657 - 65 Identification of a protein biomarker unique to the pandemic O3:K6 clone of Vibrio parahaemolyticus; Williams TL et al.; The present method of characterizing Vibrio parahaemolyticus strains involves serotyping or detection methods based on assessment of the presence or absence of genes thought to be markers of an organism's pathogenicity . It is unclear whether these assays detect all pathogenic V . parahaemolyticus strains since a clear correlation between the presence of a particular gene and the organism's pathogenicity has not yet been observed . We have described a proteomics-based method to distinguish individual V . parahaemolyticus strains on the basis of their protein profiles and identified a specific protein that is characteristic of the pandemic O3:K6 strain and its clonal derivatives . In the pandemic clone of V . parahaemolyticus, a histone-like DNA-binding protein, HU-alpha, has a C-terminal amino acid sequence different from those of other strains of V . parahaemolyticus . Upon further study, it was discovered that the gene encoding this protein has a 16-kbp insert at the 3' terminus of the open reading frame for this protein . By using the protein sequence of the unique biomarker for the pandemic clone of V . parahaemolyticus, it was possible to rationally design specific PCR-based probes and assays that permit the rapid and precise identification of pandemic strains of V . parahaemolyticus. Anal Bioanal Chem, 1996 Mar, 354(5-6), 676 - 80 Assessing toxicity and mobilisation of impregnation salts at a contaminated site; Andersen S et al.; Severe soil contamination is often encountered at wood-impregnation plants due to spills, dripping and deposition of sludge associated with dissolved salts of copper, chromium and arsenic (CCA) . Soil samples from a CCA-plant in southern Norway were analysed via a factorial extraction design to investigate mobilisation of contaminated soils . Various concentrations of organic acids, sea-salts, and pH showed that contaminants were not stable, and could be mobilised to the aqueous phase . To further investigate mobilisation of impregnation salts, soil solution collectors were installed at various depths at the site . Concentrations varied considerably . Hydrological changes revealed elevated levels of dissolved salts, which agree with the factorial experiment . Soil chemical processes (not total solid-phase concentrations) dominated the mobilisation and subsequent leaching . Soil solutions were tested for changes in toxicity by chemical analysis and degree of inhibition of luminescence in Vibrio fisheri (Microtox) . Changes in toxicity corresponded to changes in soil solution. J Biol Chem, 2004 Jun 11, 279(24), 25143 - 8 Epub 2004 Apr 05. A cellular metalloproteinase activates Vibrio cholerae pro-cytolysin; Valeva A et al.; Many strains of Vibrio cholerae produce a cytolysin (VCC) that forms oligomeric transmembrane pores in animal cells . The molecule is secreted as a procytolysin (pro-VCC) of 79 kDa that must be cleaved at the N terminus to generate the active 65-kDa toxin . Processing can occur in solution, and previous studies have described the action of mature VCC thus generated . However, little is known about the properties of pro-VCC itself . In this study, it is shown that pro-VCC exist as a monomer in solution and binds as a monomer to eukaryotic cells . Bound pro-VCC can then be activated either by exogenous, extracellular, or by endogenous, cell-bound proteases . In both cases, cleavage generates the 65-kDa VCC that oligomerizes to form transmembrane pores . A wide variety of exogenous proteinases can mediate activation . In contrast, the activating cellular protease is selectively inhibited by the hydroxamate inhibitor TAPI, and thus probable candidates are members of the ADAM-metalloproteinase family . Furin, MMP-2, MMP-9, and serine proteinases were excluded . Cells over-expressing ADAM-17, also known as tumor necrosis factor alpha converting enzyme, displayed increased activation of VCC, and knockout cells lacking ADAM-17 had a markedly decreased capacity to cleave the protoxin . The possibility is raised that pro-VCC is targeted to membrane sites that selectively contain or are accessible to cellular ADAM-metalloproteinases . Although many microbial toxins are activated by furin, this is the first evidence for processing by a cellular metalloproteinase . We identified ADAM-17 as a potent activator of pro-VCC. Appl Environ Microbiol, 2004 Apr, 70(4), 1964 - 72 Phenotypic and genotypic characteristics and epidemiological significance of ctx+ strains of Vibrio cholerae isolated from seafood in Malaysia; Chen CH et al.; Of 97 strains of Vibrio cholerae isolated from various seafoods in Malaysia in 1998 and 1999, 20 strains carried the ctx gene and produced cholera toxin . Fourteen, one, and five of these toxigenic strains belonged to the O139, O1 Ogawa, and rough serotypes, respectively . The rough strains had the rfb gene of the O1 serotype . The toxigenic strains varied in their biochemical characteristics, the amount of cholera toxin produced, their antibiograms, and the presence or absence of the pTLC plasmid sequence . DNA fingerprinting analysis by arbitrarily primed PCR, ribotyping, and a pulsed-field gel electrophoresis method classified the toxigenic strains into 3, 7, and 10 types, respectively . The relatedness of these toxigenic strains to clinical strains isolated in other countries and from international travelers was examined by using a dendrogram constructed from the pulsed-field gel electrophoresis profiles . The results of the examination of the antibiogram and the possession of the toxin-linked cryptic plasmid were consistent with the dendrogram-based relatedness: the O139 strains isolated from Malaysian seafoods could be separated into two groups that appear to have been introduced from the Bengal area independently . The rough strains of Malaysian seafood origin formed one group and belonged to a cluster unique to the Thailand-Malaysia-Laos region, and this group may have persisted in this area for a long period . The single O1 Ogawa strain detected in Malaysian seafood appears to have an origin and route of introduction different from those of the O139 and the rough strains. Pathol Biol (Paris), 2004 Apr, 52(3), 127 - 30 {Microbial polysaccharides of marine origin and their potential in human therapeutics}; Colliec-Jouault S et al.; Bacterial polysaccharides offer fascinating potential applications for the pharmaceutical industry . Although many known marine bacteria produce exopolysaccharides (EPS), continuation in looking for new polysaccharide-producing micro-organisms is promising . Marine bacteria, isolated from deep-sea hydrothermal vents, have demonstrated their ability to produce in aerobic conditions, unusual EPS . With the aim of discovering biological activities, EPS presenting different structural features were studied . An EPS secreted by Vibrio diabolicus was evaluated on the restoration of bone integrity in experimental model and was demonstrated to be a strong bone-healing material . Another EPS produc