Mol Gen Mikrobiol Virusol, 1989 Jun, (6), 3 - 6 {RecA-independence of the process of amplification caused by the RS-1 sequence of Vibrio cholerae}; Fil'kova SL et al.; The processes of amplification and deamplification of a plasmid DNA segment flanked by direct repeats of RSI-sequence of Vibrio cholerae and carrying the structural genes of cholera toxin inside the recombinant plasmid in E . coli cells have been studied . These processes determined by RSI-sequences are shown to take place independent of the RecA-system of E . coli cells. Int J Dermatol, 1989 Jun, 28(5), 311 - 6 Vibrio vulnificus infection in Hawaii; Nip-Sakamoto CJ et al.; A life-threatening Vibrio vulnificus infection occurred in a 52-year-old Korean woman with hepatic cirrhosis . Four days after ingesting raw crab, the patient presented to the hospital with nausea, vomiting, fever, hypotension, and hemorrhagic blistering of the left foot . Vibrio vulnificus was recovered from both her blood and a foot wound. Tokushima J Exp Med, 1989 Jun, 36(1-2), 41 - 5 Effects of food preservatives and local anesthetics on synthesis of outer membrane proteins in Vibrio parahaemolyticus; Koga T et al.; The effects of seven food preservatives including salicylate, benzoate, dehydroacetate, sorbate and propionate, and two local anesthetics, phenethyl alcohol and procaine, on the synthesis of major outer membrane proteins in Vibrio parahaemolyticus strain 3283-61 were investigated . The synthesis of the outer membrane proteins a, b, c and e was remarkably inhibited by all the food preservatives except propionate and two additional outer membrane proteins with respective molecular weights of about 35,000 (protein b') and 32,000 (protein c') were induced . Similar effects were encountered by the local anesthetics except that the synthesis of protein e was only weakly depressed. Southeast Asian J Trop Med Public Health, 1989 Jun, 20(2), 201 - 5 Immune responses following killed whole vibrio-B subunit oral cholera vaccine in human volunteers; Pitisuttithum P et al.; Immunogenicity of killed whole vibrio and B subunit oral cholera vaccines in American and Thai volunteers were analysed in terms of significant rise of antibody titre . Three doses of 2 x 10(11) killed vibrios and 5 mg of cholera toxin B subunit were given at two-week intervals . There were no differences in the percent of volunteers with significant rise of serum immunoglobulin G and secretory immunoglobulin A (sIgA) to cholera toxin . However, the percent with significant rises of serum antibody to whole cell V . cholerae Inaba measured by vibriocidal titre and serum immunoglobulin G, and secretory immunoglobulin A to lipopolysaccharide (LPS) measured by ELISA in American volunteers were significantly different from those in Thai volunteers (89% VS 45%, 68% VS 9% and 53% VS 0%, respectively) (p less than 0.05). Appl Environ Microbiol, 1989 Jun, 55(6), 1400 - 5 Protection of rainbow trout against vibriosis and furunculosis by the use of attenuated strains of Vibrio anguillarum; Norqvist A et al.; The fish pathogen Vibrio anguillarum causes a lethal infection in rainbow trout (Salmo gairdneri) . Three different avirulent mutants, constructed by transposon insertion mutagenesis (VAN20 and VAN70) or as antibiotic-resistant mutants (VAN1000), were isolated by screening 200 individual isolated mutants for avirulence . When used as live vaccines, all three avirulent mutants were able to induce protective immunity against the homologous as well as a heterologous strain of V . anguillarum . When VAN1000 was used, protective immunity could be recorded 1 week after bath vaccination with 10(7) bacteria per ml of water for 30 min . A single-dose immunization was effective for at least 12 weeks . Western immunoblotting showed that strains of V . anguillarum have antigenic determinants in common with Aeromonas strains . Therefore, we tested and confirmed that VAN1000 also was able to induce protective immunity against challenge with Aeromonas salmonicida. J Bacteriol, 1989 Jun, 171(6), 3549 - 52 Requirement for autoinducer in transcriptional negative autoregulation of the Vibrio fischeri luxR gene in Escherichia coli; Dunlap PV et al.; The effect of a mutation in luxI (autoinducer synthetase gene) on transcription of luxR in the cloned Vibrio fischeri lux system (luxR, luxICDABE) was examined in Escherichia coli . For the luxI mutant, transcription from the luxR promoter (monitored with beta-galactosidase levels from a luxR::lacZ fusion, with LuxR supplied in trans) decreased fivefold, to levels of the luxI+ strain, only in the presence of added autoinducer . The results demonstrate that, as has been shown at the translational level, autoinducer is required for negative autoregulation of luxR at the transcriptional level. Int J Biol Macromol, 1989 Jun, 11(3), 172 - 6 Furazolidone-induced interstrand cross-links in Vibrio cholerae DNA . Study of conformational change by circular dichroism; Chatterjee SN et al.; Vibrio cholerae DNA bearing furazolidone-induced interstand cross-links show a change in the characteristic circular dichroism spectra of the DNA itself in dilute buffer . The change in c.d . spectra was characterized by a shift of the positive band around 272 nm to lower wavelength and a loss of ellipticity of the negative band around 242 nm, and is similar to that exhibited by mitomycin linked Vibrio cholerae DNA under identical conditions and is suggestive of a conformational change of DNA bearing such cross-links . Both furazolidone-induced and mitomycin-induced cross-linking of Vibrio cholerae DNA inhibited the salt-induced conformation change, i.e . increase in winding angle of DNA, the percentage inhibition being greater for mitomycin-linked DNA. J Bioenerg Biomembr, 1989 Jun, 21(3), 347 - 57 A study on Na+ -coupled oxidative phosphorylation: ATP formation supported by artificially imposed delta pNa and delta pK in Vibrio alginolyticus cells; Dibrov PA et al.; Addition of Na+ to the K+-loaded Vibrio alginolyticus cells, creating a 250-fold Na+ gradient, is shown to induce a transient increase in the intracellular ATP concentration, which is abolished by the Na+/H+ antiporter, monensin . The delta pNa-supported ATP synthesis requires an additional driving force supplied by endogenous respiration or, alternatively, by a K+ gradient (high {K+} inside) . In the former case, ATP formation is resistant to the protonophorous uncoupler . Dicyclohexylcarbodiimide and diethylstilbestrol, but not vanadate, completely inhibit Na+ pulse-induced ATP formation . The data agree with the assumption that Na+ -ATP-synthase is involved in oxidative phosphorylation in V alginolyticus . Interrelation of H+ and Na+ cycles in bacteria is discussed. BMJ, 1989 May 20, 298(6684), 1353 - 6 Oral rehydration formula containing alanine and glucose for treatment of diarrhoea: a controlled trial; Patra FC et al.; OBJECTIVE--To determine whether adding L-alanine to the glucose based oral rehydration solution recommended by the World Health Organisation would improve its efficacy in treating acute diarrhoea . DESIGN--Randomised double blind controlled trial of oral rehydration solution containing L-alanine and glucose . SETTING--Inpatient service of a hospital treating diarrhoea . PATIENTS--97 Male patients aged 6-59 years admitted to the hospital with acute and severe dehydration due to diarrhoea associated with Vibrio cholerae or enterotoxigenic Escherichia coli . Forty nine received the standard glucose based oral rehydration solution (control group) and 48 this solution with alanine added (study group) . INTERVENTIONS--All of the patients received rapid intravenous acetate solution for the initial four hours after admission, which fully corrected the signs of dehydration . They were then admitted to the study and randomised . Immediately after the intravenous treatment oral rehydration treatment was started . All of the patients received oral tetracycline for 48 hours, starting 24 hours after start of the study . If signs of dehydration reappeared during oral treatment patients were given rapid intravenous acetate solution until they were fully corrected and then continued to take the assigned oral rehydration solution . END POINT--Passage of the last watery stool . MEASUREMENTS and MAIN RESULTS--The median stool output/kg body weight during the initial 24 hours of oral rehydration treatment and until diarrhoea stopped was reduced in the study group compared with the control group from 309 ml to 196 ml and from 393 ml to 236 ml respectively . Intake of oral rehydration solution and intravenous acetate solution was reduced from 455 ml to 308 ml and from 616 ml to 425 ml respectively . Two patients in the study group compared with 18 patients in the control group required unscheduled rapid intravenous acetate solution to correct signs of dehydration during oral rehydration treatment . CONCLUSION--Oral rehydration solution containing L-alanine was considerably better than standard oral rehydration solution at reducing the severity of symptoms and the need for fluid of male patients with diarrhoea associated with V cholerae and enterotoxigenic E coli. Mikrobiol Zh, 1989 May-Jun, 51(3), 56 - 60 {Monoclonal antibodies to antigens of cholera vibrios of the Ogava serovar}; Burlakhova OS et al.; A set of hybrids is obtained synthesizing monoclonal antibodies to the surface antigenic determinants of the choleric vibrio of the Ogava serovar . The antigenic structure of vibrios of the typical and atypical strains isolated from a man and from environmental objects is studied using a collection of highly specific immunoglobulins . The high-specific diagnostic preparations for identification of the 01-group cholera agent serovar may be created on their base. J Biochem (Tokyo), 1989 May, 105(5), 723 - 7 Differential exposure of the major gangliosides in rabbit thymocytes to Vibrio cholerae neuraminidase; Iwamori M et al.; Neuraminidase treatment of lymphocytes is known to cause changes of cellular responses in several biological phenomena, but the molecules modified on the cell surface by neuraminidase are not known in detail . Rabbit thymocytes, which contain tissue-characteristic gangliosides, were treated with Vibrio cholerae neuraminidase, and the susceptibility of the cell surface sialic acid residues was examined . The amount of sialic acid released from the thymocytes at the highest level was 42.4 nmol per 1 X 10(9) cells, among which 26.5% was from gangliosides . Ninety-three percent of the VI3NeuGc-nLc6Cer, 84% of the IV3NeuGc-nLc4Cer, and 50% of the II3NA2-LacCer in the thymocytes was hydrolyzed to nLc6Cer, nLc4Cer, and LacCer, respectively, but II3NA-LacCer was completely cryptic . Also, among the molecular species of II3NA2-LacCer, C20:0- to C24:0-containing, but not C16:0- to C18:0-containing molecules, were susceptible to neuraminidase . After neuraminidase treatment, nLc4Cer and nLc6Cer became the major glycosphingolipids, and a 15-fold increase of radioactivity incorporated into the glycosphingolipids was observed by the galactose oxidase-sodium borotritide procedure, suggesting that the beta-galactose of the glycosphingolipids produced by neuraminidase treatment is accessibly to the several ligands which are functionally associated with lymphocytes. Indian J Med Res, 1989 May, 89, 128 - 31 Virulence of Vibrio vulnificus strains isolated from seafoods; Malathi GR et al.; A study was carried out on the relation between virulence of V . vulnificus assessed by LD50 in mice and other markers . V . vulnificus strains isolated from seafoods had LD50 ranging from 10(3)-10(7) for mice . Strains showing LD50 10(6) or less were generally haemolytic and settled after boiling . Resistance to bactericidal action of serum and ability to take up congo red was shown by all the strains. Indian J Med Res, 1989 May, 89, 117 - 20 Neutralisation of the new cholera toxin by antiserum against crude enterotoxin of cholera toxin gene-positive Vibrio cholerae 01 in rabbit ileal loop model; Saha S et al.; The enterotoxicity of the new cholera toxin (NCT) prepared from cholera toxin gene-negative (CT-) V . cholerae 01 strains isolated from human diarrhoeal and environmental sources was assayed in rabbit ileal loops and the toxin unit was calculated to be 24 micrograms of protein . The enterotoxicity of the NCT preparations were completely neutralised by the antiserum raised against the enterotoxin preparation from the CT+ V . cholerae 01 strain 569B at 1 in 16 dilution in ileal loops . The antiserum contained 1 unit of antitoxin in 85 x 10(-4) ml amount . The data indicate that the antiserum prepared against the enterotoxin of CT+ strain contains antibody against the NCT and can neutralise the toxin in vivo . The observations also suggest that CT+ strains liberate the NCT simultaneously with CT and the latter gets eliminated during the process of purification. Salud Publica Mex, 1989 May-Jun, 31(3), 314 - 25 {Prevalence of Vibrio parahaemolyticus in seafood from restaurants in the city of Mérida, Yucatán}; Franco-Monsreal J et al.; We report the prevalence of the Vibrio parahaemolyticus in raw (11.54%), insufficiently cooked (0.00%) and partially cooked with heat (4.55%) sea foods, from restaurants of the city of Merida, Yucatan, Mexico . We studied 190 samples from March to August 1987 . We isolated 12 strains (6.32%) from an equal number of samples, whose biochemical characteristics corresponded with Vibrio parahaemolyticus . We didn't find significant statistically differences when we compared the prevalences of the samples of raw and insufficiently cooked sea foods (chi 2 = 0.03, p greater than 0.05), of raw and partially cooked with boiled sea foods (chi 2 = 1.96, p greater than 0.05), and of insufficiently heated and partially cooked with heat sea foods (chi 2 = 0.24, p greater than 0.05) . When we compared the prevalences of the samples of crustaceans and mollusks, of crustaceans and fishes, and of mollusks and fishes, we didn't find significant statistically differences: chi 2 = 0.022, p greater than 0.05; chi 2 = 0.52, p greater than 0.05 and chi 2 = 1.78, p greater than 0.05, respectively. Photochem Photobiol, 1989 May, 49(5), 681 - 8 Lux C, D and E genes of the Vibrio fischeri luminescence operon code for the reductase, transferase, and synthetase enzymes involved in aldehyde biosynthesis; Boylan M et al.; The lux C, D, and E genes of the Vibrio fischeri luminescence operon code for three polypeptides of 54, 33, and 42 kDa, respectively, which are required for synthesis of the aldehyde substrate for the luminescent reaction . These polypeptides have been identified in V . fischeri and V . harveyi as well as in recombinant E . coli harboring the cloned genes by specific acylation with {3H}fatty acid, showing that they are components of a fatty acid reductase system with reductase, synthetase and transferase activities . By using glycerol in the assay and/or extraction buffer and decreasing the reducing agent, the levels of the acylation of the 54 and 42 kDa polypeptides have been greatly increased . As a consequence, it was possible to demonstrate that the 54 kDa polypeptide coded by the lux C gene has reductase activity . In a subclone missing the lux E gene, the 42 kDa polypeptide was missing and the 54 kDa polypeptide could not be acylated in vitro with tetradecanoic acid (+ATP) and only to a low level in vivo indicating that the synthetase enzyme, responsible for fatty acid activation, is coded by the lux E gene . In vitro acylation with tetradecanoyl CoA of the 33 kDa polypeptide coupled with the specific cleavage of acyl-ACP only in E . coli extracts transformed with DNA containing the lux D gene, demonstrated that the lux D gene coded for the transferase enzyme. J Med Microbiol, 1989 May, 29(1), 33 - 9 Identification of some antigenically related outer-membrane proteins of strains of Vibrio cholerae O1 and non-O1 serovars involved in intestinal adhesion and the protective role of antibodies to them; Sengupta D et al.; Outer-membrane proteins (OMPs) of Vibrio cholerae strains of O1 and non-O1 serovars were studied . Marked similarity was found in the OMP profiles of different V . cholerae O1 strains but the OMP profile of a non-O1 strain was somewhat different . Antigenic relatedness between the OMPs of different V . cholerae strains was established by enzyme-linked immunosorbent assay (ELISA) . Immunoblotting experiments demonstrated that at least two OMPs of 36 and 25-26 Kda were immunogenic and common to strains of O1 and non-O1 serovars . Antiserum raised against the outer membrane of a V . cholerae strain, and rendered specific for its OMP by absorption with lipopolysaccharide, inhibited in vitro the intestinal adhesion of the homologous and heterologous strains of V . cholerae irrespective of their biotype, serotype and serovar . Furthermore, antiserum to OMPs induced passive protection against vibrio challenge in rabbit ileal loop experiments . These results suggest that the OMPs may be useful in immunoprophylaxis against cholera. J Ky Med Assoc, 1989 May, 87(5), 219 - 22 Vibrio vulnificus sepsis after eating raw oysters; Stahr B et al.; Vibrio vulnificus, a marine vibrio found in coastal waters of the United States, may contaminate certain seafoods, particularly raw oysters . Patients with underlying liver disease are particularly susceptible to severe illness . Unexplained febrile diseases in patients who have eaten raw oysters may be caused by Vibrio vulnificus . A fatal and a nonfatal case are reported in two such patients . These patients are the first two reported cases of Vibrio vulnificus infection in Kentucky. J Histochem Cytochem, 1989 May, 37(5), 663 - 74 Immunogold labeling of luciferase in the luminous bacterium Vibrio harveyi after fast-freeze fixation and different freeze-substitution and embedding procedures; Nicolas MT et al.; We studied the ultrastructural localization of luciferase on sections of the bioluminescent bacterium Vibrio harveyi by indirect immunogold staining, using a polyclonal antiluciferase antibody and the usual control tests, after chemical fixation or fast-freeze fixation (FFF) followed by different freeze-substitution (FS) procedures and embedding in either Epon or LR White . After liquid fixation with glutaraldehyde and paraformaldehyde and LR White embedding, labeling occurred over the cytoplasm but not over the condensed nucleoid . Epon embedding almost abolished it . FFF-FS considerably improved the morphological preservation and revealed cytoplasmic "patches" with a complex ultrastructure in Epon sections . The preservation was always less good in LR White . The patches were densely labeled, even in Epon sections, after FS in acetone . However, labeling intensity was 3.7 times greater in LR White than in Epon . With both resins, labeling diminished similarly when fixative agents were present in the FS medium . The localization of luciferase in the cytoplasm and particularly in the patches is discussed. Appl Environ Microbiol, 1989 May, 55(5), 1178 - 86 Characterization of the microbial community colonizing the anal and vulvar pores of helminths from the hindgut of zebras; Mackie RI et al.; Scanning and transmission electron microscopy were used to examine the adherence and in situ morphology of the microbial community colonizing the anal and vulvar pores of the subfamily Cyathostominae (Nematoda: Strongylidae) from the colon of Burchell's zebra (Equus burchelli antiquorum) . Two different morphological types of asporogenous rod were prominent in the microbial community . One was a thin, septate, filamentous organism (0.4 to 0.5 micron by 2 to 3 microns) with blunt ends, which was more prominent at the site of attachment . The other was a larger (1.8 to 2.4 microns by 5 to 10 microns) multicellular rod with round ends in the form of a trichome . Spiral- and vibrio-shaped bacteria were also present in the thin sections . The septate filaments were shown to contain a cell spacer similar to those described in Methanospirillum hungatei . Attachment to the cuticle was by means of an amorphous electron-dense material with fibrillar appearance and not by specialized holdfast segments . Ten isolates were obtained from a habitat-simulating medium on which a homogenate from the posterior region was plated . Antibodies were raised to whole cells of five rod-shaped isolates in rabbits and fluorescein isothiocyanate labeled . Positive bright-yellow fluorescence was obtained with one of the clostridial isolates . The results are discussed with reference to other bacteria with similar morphology, the nature of this unique interrelationship between the microbial community and its parasitic host inside the equine hindgut, and the possibility of biological control of parasitic helminths. J Exp Zool, 1989 May, 250(2), 219 - 28 Bacterial action of fertilization envelope extract from eggs of the fish Cyprinus carpio and Plecoglossus altivelis; Kudo S et al.; The vitelline envelope (VE) and fertilization envelope (FE) in eggs of the fish Cyprinus carpio and Plecoglossus altivelis were purified by homogenization of eggs or embryos in 5 mM Tris-HCl buffer, pH 7.0, containing 2 mM ethylenediamine tetraacetic acid disodium salt (EDTA), except for processing of VEs in Plecoglossus eggs, and by repeated washing wih the same buffer . To extract the outermost layer material, the purified VEs and FEs were processed overnight at 4 degrees C in 5 mM Tris-HCl buffer, pH 7.0, containing 8 mM 2-mercaptoethanol, 2 mM EDTA, 0.3 M alpha-lactose, 0.3 M glucose, and 0.9% NaCl . Since extraction of the outermost layer of the VEs of Cyprinus eggs in this solution was found to be ultrastructurally incomplete, further sonication in the same buffer was necessary . The solution extracted from purified VEs or FEs was dialyzed against 5 mM Tris-HCl buffer, pH 7.0, followed by lyophilization . The extracts from the FEs from both fish species contained two kinds of lectins, one agglutinated human B-type erythrocytes and the other nonspecifically agglutinated fish spermatozoa, and both extracts had a strong bactericidal effect on Vibrio anguillarum that was isolated from diseased cultured fish, but not on Aeromonas hydrophila and Escherichia coli . The extracts of purified VEs from eggs of both fish had no bactericidal effect on the bacteria examined, nor any agglutination effect on human erythrocytes and fish spermatozoa. J Bacteriol, 1989 May, 171(5), 2293 - 302 Identification of polypeptides encoded by cloned pJM1 iron uptake DNA isolated from Vibrio anguillarum 775; Singer JT et al.; The XhoI fragment containing much of the iron uptake region of plasmid pJM1 was isolated from Vibrio anguillarum 775 and cloned into plasmid pBR322 . Plasmid-encoded polypeptides were examined in maxicells of Escherichia coli, and transposon mutagenesis was used to map insertion mutations in the structural DNA encoding the OM2 polypeptide . Tn1000 insertions that mapped within OM2 and blocked maxicell expression of OM2 resulted in the loss of ferric iron-anguibactin receptor function when plasmids containing OM2:: Tn1000 insertions were introduced into V . anguillarum cells . Two iron-regulated polypeptides were identified in maxicell polypeptide profiles of E . coli SS201 . A 20,000-dalton polypeptide was expressed in maxicells of SS201 grown under conditions of iron limitation but was barely detectable in profiles of SS201 cells that were grown under high-iron conditions . DNA encoding the 20,000-dalton polypeptide mapped downstream of and adjacent to the gene encoding OM2 . DNA sequences required for production of a 46,000-dalton polypeptide mapped 4.5 kilobases downstream of the OM2 structural gene . The 46,000-dalton polypeptide was synthesized at high levels in E . coli SS201 maxicells grown under high-iron conditions, but synthesis of the protein was severely repressed under conditions of iron limitation . Iron-regulated expression of both proteins in maxicells of SS201 was relieved upon deletion of a 4.9-kilobase SalI-XhoI fragment of pJM1 DNA, which indicated that pJM1 DNA sequences present in the deleted fragment are required for regulated expression of both proteins in E . coli . Maxicells of SS201 harboring these deletion derivatives synthesized the 20,000-dalton polypeptide at very low constitutive levels and the 46,000-dalton polypeptide at high constitutive levels, regardless of the iron concentration of the growth medium . The observed regulation of the 20,000-dalton protein suggested that it might play a role either in siderophore biosynthesis or in the functional expression of OM2 . The opposite regulatory pattern observed for the 46,000-dalton polypeptide suggested that it does not play a structural role in siderophore or OM2 biosynthesis, but the observed regulatory pattern might be expected if the 46,000-dalton protein played a negative regulatory role in siderophore biosynthesis. J Clin Microbiol, 1989 May, 27(5), 1015 - 21 Detection of piluslike structures on clinical and environmental isolates of Vibrio vulnificus; Gander RM et al.; Twenty clinical isolates of Vibrio vulnificus were compared with 10 environmental strains by using electron microscopy and agglutination assays with human erythrocytes, guinea pig erythrocytes, and Saccharomyces cerevisiae . In addition, the isolates were tested for ability to adhere to the human epithelial cell lines HEp-2 and A549 . When examined by electron microscopy, 16 (80%) of the 20 clinical isolates demonstrated the presence of piluslike structures; the composition of the bacterial populations ranged from 0 to 68% piliated cells . In contrast, only 3 (30%) of the 10 environmental isolates were piliated, with a range from 0 to 16% piliated cells . A significant association between the presence of piliated cells and the isolate source was found (P less than 0.05) . None of the 30 strains agglutinated erythrocytes or yeast cells . V . vulnificus adherence results obtained with HEp-2 cells showed 10 (50%) of 20 clinical isolates and 0 (0%) of 10 environmental isolates with averages of greater than 10 adherent bacteria per cell, demonstrating a correlation between attachment and the isolate source (P less than 0.05) . Selected strains were tested to determine whether methyl alpha-D-mannopyranoside, fructose, or alpha-L-(-)-fucose would inhibit bacterial adherence to HEp-2 cells . Multiple patterns of adherence inhibition were observed . Adherence to A549 cells showed 8 (40%) of 20 clinical isolates and 0 (0%) of 10 environmental strains with averages of greater than 10 adherent bacteria per cell . A statistical association between attachment and the isolate source was demonstrated (P less than 0.05) . These data suggest that the presence of piluslike structures and the ability to adhere to human epithelial cell lines may be more closely associated with V . vulnificus isolates from clinical specimens than with environmental strains. Exp Hematol, 1989 May, 17(4), 351 - 6 Effects of neuraminidase on the regulation of erythropoiesis: III: Characterization of carbohydrate moieties on the surface of thymic regulatory cells that interact with erythroid colony-forming cells; La Russa VF et al.; In this study we further define cell surface carbohydrate structures relevant to cellular interactions that regulate erythropoiesis . An analysis of thymocyte cell surface negativity was made using fluoresceinated poly-L-ornithine (FITC poly-L-ornithine) as a probe that binds to negatively charged sites (i.e., sialic acid residues) at the cell surface . Two distinct subpopulations are labeled, comprising both intensely as well as weakly fluorescent subpopulations of thymocytes . Prior treatment of thymocytes with Vibrio cholerae neuraminidase (VCN), which removes cell surface sialic acid residues, markedly reduced the FITC poly-L-ornithine surface labeling of these cells . Distinct enzymatic modifications of regulatory cell functions were also assessed by the ability of thymocytes to function as separate regulatory subpopulations . Confirming our previous observations, treating thymocytes with VCN impaired the enhancement activity but had little effect on thymocyte regulatory ability to suppress erythroid colony growth . In contrast, treatment of thymocytes with galactose oxidase (GAO) or beta-galactosidase (beta-GAL) removed suppressor activity either before or after VCN treatment . A further exposure of GAO-treated thymocytes to sodium borohydride or hydroxylamine, which reduce D-galactose residues, restores their suppressor function and prevents enhancement . These differential enzymatic effects on thymocyte regulatory cell functions suggest that different carbohydrate structures may be involved in helper and suppressor activities for erythroid colony formation . Sialic acid residues may be associated with certain cells that function to enhance erythropoiesis, and D-galactose residues may be associated with the suppressor subpopulation. Mol Immunol, 1989 May, 26(5), 485 - 94 Interleukin-1 induction by lipopolysaccharides: structural requirements of the 3-deoxy-D-manno-2-octulosonic acid (KDO); Haeffner-Cavaillon N et al.; We previously showed the importance of the 3-deoxy-D-manno-2-octulosonic acid (KDO) residue in endotoxins (lipopolysaccharides, LPS) for the induction of the synthesis and release of interleukin-1 (IL-1) by human monocytes . We further investigated the effect of some structural variations within the KDO molecule on IL-1 production induced by LPS . Deamination of Bordetella pertussis LPS, followed by mild anhydrous acidic methanolysis released a hexasaccharide (fragment B'), which had a terminal methyl ketoside KDO residue with a methyl-esterified carboxyl group . This fragment was unable to induce IL-1 production by human monocytes . Fragment B' could be converted into an active hexasaccharide by de-esterification (fragment B-OMe), but not by reduction of the methyl ester group . The KDO residues in the LPS of some bacterial species have been shown to be phosphorylated and we observed that these LPS were weak IL-1 inducers . Phosphorylated KDO present in Vibrio cholerae and B . pertussis LPS respond poorly in the thiobarbiturate assay (specific for KDO) . However, if these LPS were dephosphorylated with aqueous hydrofluoric acid (HF) their KDO response in this assay was increased 5.4- to 2.6-fold, respectively . In parallel, the HF-treated LPS were more potent IL-1 inducers than untreated endotoxins . These data confirm that the KDO residue(s) present in all endotoxins play(s) a major role in the signal(s) leading to IL-1 production by human monocytes, and show that IL-1 induction by LPS (1) requires a free carboxyl group in the KDO and (2) is correlated with the degree of substitution of the KDO. J Biochem (Tokyo), 1989 May, 105(5), 841 - 6 Cloning and expression of the 5'-nucleotidase gene of Vibrio parahaemolyticus in Escherichia coli and overproduction of the enzyme; Sakai Y et al.; The gene encoding the membrane-bound 5'-nucleotidase of Vibrio parahaemolyticus was cloned and expressed in Escherichia coli . Cells of E . coli harboring a plasmid, pNUT5, which carries the 5'-nucleotidase gene were able to grow on ATP as the sole source of carbon, although the original cells were not . The 5'-nucleotidase activity was detected in whole cells of E . coli harboring pNUT5 and in membrane vesicles prepared from these cells . Most properties of the 5'-nucleotidase produced in E . coli, that is, its requirements for Cl- and Mg2+, substrate specificity, and inhibition by Zn2+, were similar to those observed in V . parahaemolyticus, but some alterations in properties were observed: The 5'-nucleotidase was partially inducible in V . parahaemolyticus, but its expression in E . coli was completely constitutive . The specific activity of the 5'-nucleotidase in membrane vesicles of E . coli harboring the plasmid was 30 times that observed in whole cells, whereas the specific activities in membrane vesicles and in whole cells of V . parahaemolyticus were almost the same . A new, dense band of protein with an apparent molecular mass of 63 kDa was detected when membrane proteins of E . coli harboring the plasmid were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Biochem (Tokyo), 1989 May, 105(5), 785 - 9 Purification and characterization of membrane-bound 5'-nucleotidase of Vibrio parahaemolyticus; Itami H et al.; Membrane-bound 5'-nucleotidase from Vibrio parahaemolyticus was solubilized and purified using a nonionic detergent, heptyl-beta-D-thioglucoside, and was characterized . This enzyme required Mg2+ for activity, maximum activity being observed at 5 and 20 mM Mg2+ with AMP and ATP, respectively, as substrates . Of the divalent cations tested, Mn2+ and Co2+ were able to replace Mg2+ partially, whereas Ca2+ was ineffective . Zinc strongly inhibited the enzyme activity and Ni2+ caused partial inhibition . This enzyme required Cl- for activity, the optimal concentration being 20 mM or more . The order of effectiveness of anions was Cl- greater than Br- greater than I- approximately NO3- . Sulfate and acetate were ineffective . The optimal pH was 8.0 . The activity of the purified enzyme was stimulated by the addition of lipid to the assay mixture . This enzyme hydrolyzed all 5'-nucleotides tested, but did not hydrolyze 3'-nucleotides or ribose 5-phosphate . On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme appeared to be a single polypeptide, with a molecular weight of 72 kDa. Proc Natl Acad Sci U S A, 1989 May, 86(10), 3529 - 33 Regulation of the iron uptake system in Vibrio anguillarum: evidence for a cooperative effect between two transcriptional activators; Salinas PC et al.; We have identified a 110-kDa polypeptide that has a trans-acting regulatory activity on the expression of the pJM1 plasmid iron-uptake genes in Vibrio anguillarum . This protein is encoded by the angR gene and maps in a 3.6-kilobase-pair pJM1 DNA region located downstream of the iron transport genes . Full expression of this gene occurs under iron-limiting conditions and requires a 2.9-kilobase-pair upstream region in cis that maps within the coding region of the OM2 outer membrane protein, essential for the transport of iron into the cell cytosol . Determination of the siderophore anguibactin levels as well as analysis of specific transcripts for anguibactin biosynthetic genes demonstrated that AngR and another transcriptional activator, Taf, regulate in a synergistic fashion the level of anguibactin production by activation of transcription of the anguibactin biosynthetic genes under iron-limiting conditions. J Bacteriol, 1989 May, 171(5), 2406 - 14 Identification of a locus controlling expression of luminescence genes in Vibrio harveyi; Martin M et al.; Mutagenesis with transposon mini-Mulac was used to identify loci containing genes for bioluminescence (lux) in the marine bacterium Vibrio harveyi . Transposon insertions which resulted in a Lux- phenotype were mapped to two unlinked regions of the genome . Region I contained the luxCDABE operon which was previously shown to encode the enzymes luciferase and fatty acid reductase, which are required for light production . The other locus, region II, which was identified for the first time in this study, appeared to have a regulatory function . In Northern blot analysis of mRNA from mutants with defects in this region, no transcription from the luxCDABE operon could be detected . Strains with transposon-generated lux::lacZ gene fusions were used to analyze control of the transcription of these regions . Expression of luminescence in the wild type was strongly influenced by the density of the culture, and in strains with the lacZ indicator gene coupled to the luxCDABE operon, beta-galactosidase synthesis was density dependent . So, transcription of this operon is responsive to a density-sensing mechanism . However, beta-galactosidase synthesis in strains with lacZ fused to the region II transcriptional unit did not respond to cell density . The organization and regulation of the lux genes of V . harveyi are discussed, particularly with regard to the contrasts observed with the lux system of the fish light-organ symbiont Vibrio fischeri. Dtsch Zahnarztl Z, 1989 May, 44(5), 378 - 9 {Vital staining and phase contrast microscopy results of plaque smears}; Muller RF et al.; The information about the vitality of the plaque is limited . Only motile bacteria can be distinguished as alive . Vital staining allows to differentiate between living nonmotile rods and dead, previously motile rods . With increasing severity of the periodontal destruction, the proportion of alive respectively motile rods and vibrions increases . The vital staining shows very clearly the relation between destruction and living rods. J Gen Microbiol, 1989 May, 135 ( Pt 5), 1195 - 200 Variation in epitopes of the B subunit of El Tor and classical biotype Vibrio cholerae O1 cholera toxin; Tamplin ML et al.; Variation in epitopes of the B subunit of cholera toxin (CT-B) produced by strains of El Tor and classical biotype Vibrio cholerae O1 was examined using monoclonal antibodies prepared to V . cholerae 569B CT . CT-B epitopes were markedly conserved for V . cholerae classical biotypes . In contrast, epitope variation was observed for El Tor biotypes, which produced both a classical-like CT-B and a unique CT-B lacking at least one epitope common to 569B CT-B . The missing epitope was located outside the GM1 ganglioside-binding site . From results of the study reported here, genetic divergence is exhibited in the El Tor biotype CT-B versus classical CT-B . Furthermore, at least five unique epitopes of V . cholerae 569B CT-B can be defined. Infect Immun, 1989 Apr, 57(4), 1227 - 34 Bovine lactogenic immunity against cholera toxin-related enterotoxins and Vibrio cholerae outer membranes; Boesman-Finkelstein M et al.; The newly parturient cow secretes large quantities of immunoglobulin G1, a relatively protease- and heat-resistant immunoglobulin, in its colostrum and milk . This study establishes the feasibility of producing protective colostral immunoglobulins by immunizing pregnant cows with cholera toxin (CT), a CT-related enterotoxin from Escherichia coli, and Vibrio cholerae outer membranes (OMs) . The OMs were prepared from bacteria grown under iron-replete or iron-deficient (to simulate the in vivo environment) conditions . Immunoglobulins were purified from the colostrum of newly parturient control and immunized cows . The bovine anti-CT and anti-H-LT (CT-related heat-labile enterotoxin produced by diarrheogenic E . coli strains of human origin) antibodies were quantitated by enzyme-linked immunosorbent assays and by neutralization of toxin activity in both Y-1 adrenal cell and infant rabbit assays . The bovine anti-OM antibodies from both high-iron-grown and low-iron-grown vibrios were assessed by bacterial agglutination and by Western blot (immunoblot) analysis of polyacrylamide gel electrophoresis of high-iron-grown and low-iron-grown OMs . To test their protective effect, immunoglobulin preparations were administered orally in infant feeding formula to 6-day-old rabbits . Anti-CT and anti-OM immunoglobulins elicited statistically significant protection against diarrhea in infant rabbits challenged intraintestinally with virulent cholera vibrios. Appl Environ Microbiol, 1989 Apr, 55(4), 826 - 31 Relationships between plasmids and phenotypes of presumptive strains of Vibrio anguillarum isolated from different fish species; Wiik R et al.; On the basis of plasmid composition as well as serological and biochemical properties, 26 strains identified as Vibrio anguillarum isolated from diseased fish could be assigned to two different groups . Except for three reference strains, these++ strains were isolated from Norwegian fish . The four strains isolated from rainbow trout (Salmo gairdneri), the only strain isolated from char (Salvelinus alpinus), and three of six strains isolated from Atlantic salmon (Salmo salar) harbored a plasmid of 47 megadaltons (MDa) . Restriction endonuclease analysis showed that this plasmid and the virulence plasmid pJM1, carried by V . anguillarum strain 775, were very similar but not identical . Strains harboring the 47-MDa plasmid had nearly identical biochemical properties and were serotype O1 . Strains isolated from reared coastal cod (Gadus morhua), turbot (Scophthalmus maximus), halibut (Hippoglossus hippoglossus), free-living saithe (Pollachius virens), and partly from reared Atlantic salmon differed from strains harboring the 47-MDa virulence plasmid by not containing this plasmid, by having different biochemical traits, and by being serotype O2 . Rainbow trout which were experimentally infected with a strain isolated from cod suffering from vibriosis developed clinical symptoms similar to those in cod but quite different from those usually seen in rainbow trout. Appl Environ Microbiol, 1989 Apr, 55(4), 819 - 25 Virulence studies based on plasmid profiles of the fish pathogen Vibrio salmonicida; Wiik R et al.; Strains of Vibrio salmonicida isolated from Atlantic salmon (Salmo salar) and rainbow trout (Salmo gairdneri) suffering from cold-water vibriosis could be divided on the basis of plasmid profiles into four different categories . Of 32 strains, 19% harbored three plasmids of 24, 3.4, and 26 megadaltons (MDa), 69% harbored the 24- and 3.4-MDa plasmids but not the 2.6-MDA plasmid, and 9% harbored only the 24-MDA plasmid . The fourth category, which consisted of only one strain, harbored a plasmid of 10 MDa . In spite of different plasmid patterns, the strains of V . salmonicida were very similar with respect to biochemical reactions . The one-third of the V . salmonicida strains which were serotyped were of the same type . The 50% lethal doses, which were determined by intraperitoneal injection, ranged from 4 x 106 to 1 x 108 CFU per fish. J Med Microbiol, 1989 Apr, 28(4), 287 - 90 Bacterial growth and toxin production in ileostomy effluents; Ala Aldeen DA et al.; Escherichia coli (2), Vibrio cholerae (2) and Aeromonas sobria (1) strains were examined for their ability to grow and produce toxins in samples of ileostomy fluid . Three categories of response were observed: no detectable growth, growth without detectable toxin, and growth with detectable toxin . Clear differences were apparent between samples of ileostomy fluid obtained from different individuals and between samples obtained from the same individual at different times . The patterns of response were unique for each of the five test strains . We propose that the procedure developed forms a basis for investigating the host-parasite relationship in diarrhoeal disease. Rev Argent Microbiol, 1989 Apr-Jun, 21(2), 71 - 7 {Isolation of Vibrio cholerae non 01 from sewage in Argentina}; Corrales MT et al.; The Vibrio cholerae non 01 closely related to the classic choleric vibrio epidemic has acquired worldwide importance during the last decade, with outbreaks of diarrheas, septicemia and other disorders in humans and animals . Contaminated food and water and also liquids from sewers offer important steps in the transmission chain . Its isolation in the Atlantic and Pacific coast has led us to investigate its presence in our country, using sewage for the first study . We isolated for the first time in Argentina 27 strains of Vibrio cholerae non 01 from samples taken in the district of Berisso which discharges its waters into the River Plate. J Bacteriol, 1989 Apr, 171(4), 1870 - 8 Broad-host-range vectors for delivery of TnphoA: use in genetic analysis of secreted virulence determinants of Vibrio cholerae; Taylor RK et al.; Gene fusions between the cholera toxin structural genes and phoA, which encodes bacterial alkaline phosphatase, were identified after TnphoA mutagenesis of the cloned genes in Escherichia coli and were then mobilized into Vibrio cholerae . The activities of the hybrid proteins were detectable in V . cholerae and suggested that, like cholera toxin, they were secreted beyond the cytoplasm . To extend the utility of TnphoA to identify additional genetic export signals in V . cholerae and other gram-negative bacteria, TnphoA delivery vectors utilizing broad-host-range plasmids were developed . By using V . cholerae as a model system, insertion mutants carrying active phoA gene fusions were identified as colonies expressing alkaline phosphatase, which appeared blue on agar containing the indicator 5-bromo-4-chloro-3-indolyl phosphate . Since alkaline phosphatase is active only upon export from the cytoplasm, PhoA+ colonies resulting from the mutagenesis procedure were enriched for insertions in genes that encode secreted proteins . Insertion mutations were identified in the gene encoding a major outer membrane protein, OmpV, and in tcpA, which encodes a pilus (fimbrial) subunit . Mutant strains harboring chromosomal insertions isolated in this manner can be used to assess the role of the corresponding inactivated gene products on survival of V . cholerae in vivo . The expression of the hybrid proteins as determined by measuring alkaline phosphatase activity also allowed the convenient study of virulence gene expression. J Bacteriol, 1989 Apr, 171(4), 1825 - 34 Establishment of gene transfer systems for and construction of the genetic map of a marine Vibrio strain; Ichige A et al.; Two gene transfer systems were established for a marine bacterium, Vibrio sp . strain 60 . One was generalized transduction with a newly isolated bacteriophage, As3, and the other was conjugal gene transfer by the use of newly constructed transposon-facilitated recombination (Tfr) donors . As3 transduced various chromosomal markers at frequencies of 10(-4) to 10(-6) . Tfr donors, which were constructed by introducing transposon Tn10 into both plasmid RP4 and the chromosome, mediated the polarized transfer of chromosomal genes from the sites of Tn10 insertion on the chromosome . By means of these gene transfer systems, a genetic map of the vibrio chromosome was constructed. Biochim Biophys Acta, 1989 Mar 23, 973(3), 450 - 6 A respiratory-driven and an artificially driven ATP synthesis in mutants of Vibrio parahaemolyticus lacking H+-translocating ATPase; Sakai Y et al.; Mutants of Vibrio parahaemolyticus lacking the H+-translocating ATPase were isolated to evaluate both the role of this enzyme and the possibility of the involvement of other cation-translocating ATPase in the energy transduction in this organism . Dicyclohexylcarbodiimide-sensitive ATPase activity which represents the H+-translocating ATPase was not detected either in the membrane vesicles or in the cytosol of the mutants . Three major subunits, alpha, beta and gamma, of the H+-translocating ATPase were missing in the membranes of the mutants . Although ATP was synthesized in wild type cells when an artificial H+ gradient was imposed, little ATP was synthesized in the mutants . However, we observed a large ATP synthesis driven by the respiration not only in the wild type but also in the mutants . The respiratory-driven ATP synthesis in wild type was inhibited by an H+ conductor, carbonylcyanide m-chlorophenylhydrazone, by about 50% . On the other hand, the ATP synthesis in the mutants was not affected by the H+ conductor . Since this organism possesses a respiratory Na+ pump, Na+-coupled ATP synthesis might take place . In fact, we observed some ATP synthesis driven by an artificially imposed Na+ gradient both in the wild type and the mutant. Biochemistry, 1989 Mar 21, 28(6), 2684 - 9 Random and site-directed mutagenesis of bacterial luciferase: investigation of the aldehyde binding site; Chen LH et al.; Numerous luciferase structural gene mutants of Vibrio harveyi have been generated by random mutagenesis and phenotypically characterized {Cline, T.W., & Hastings, J.W . (1972) Biochemistry 11, 3359-3370} . All mutants selected by Cline and Hastings for altered kinetics in the bioluminescence reaction had lesions in the alpha subunit . One of these mutants, AK-20, has normal or slightly enhanced thermal stability and enhanced FMNH2 binding affinity but a much-reduced quantum yield of bioluminescence and dramatically altered stability of the aldehyde-C4a-peroxydihydroflavin-luciferase intermediate (IIA), with a different aldehyde chain length dependence from that of the wild-type luciferase . To better understand the structural aspects of the aldehyde binding site in bacterial luciferase, we have cloned the luxAB genes from the V . harveyi mutant AK-20, determined the nucleotide sequence of the entire luxA gene, and determined the mutation to be TCT----TTT, resulting in a change of serine----phenylalanine at position 227 of the alpha subunit . To confirm that this alteration caused the altered kinetic properties of AK-20, we reverted the AK-20 luxA gene by oligonucleotide-directed site-specific mutagenesis to the wild-type sequence and found that the resulting enzyme is indistinguishable from the wild-type luciferase with respect to quantum yield, FMNH2 binding affinity, and intermediate IIA decay rates with 1-octanal, 1-decanal, and 1-dodecanal . To investigate the cause of the AK-20 phenotype, i.e., whether the phenotype is due to loss of the seryl residue or to the properties of the phenylalanyl residue, we have constructed mutants with alanine, tyrosine, and tryptophan at alpha 227.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1989 Mar, 57(3), 969 - 74 Binding of cholera toxin to pig intestinal mucosa glycosphingolipids: relationship with the ABO blood group system; Bennun FR et al.; A search for compounds from intestinal mucosa of pigs carrying and not carrying blood group A-active substances (A+ and A- pigs, respectively) capable of binding cholera toxin (CT) was performed . Glycolipid extracts from a pool of pig intestinal mucosa resolved in thin-layer chromatography (TLC) revealed the presence of six to eight compounds capable of binding 125I-CT, two of them running as the ganglioside standards GM1 and GD1b . When intestinal mucosa glycolipids from single pigs were assayed by TLC for CT-binding capacity, two different patterns of labeling were observed . The main difference was at the level of compounds running below GD1b . The A+ pigs but not the A- pigs showed CT binding at this level . The major CT-binding compound detected only in A+ pigs was purified and some properties were determined . After TLC developed with different solvent systems, the purified compound bound CT and also immunoreacted with anti-A and anti-AB antisera but not with anti-B antiserum . The compound was also able to inhibit the hemagglutination of human A erythrocytes caused by anti-A antiserum, but inhibition was not observed with the B-anti-B or O (H)-Ulex europaeus lectin systems . A partial chemical characterization indicated that the active compound is a neutral glycosphingolipid containing glucose, fucose, galactose, and hexosamine . The existence of a blood group-active substance(s) able to interact with CT may help to explain the relationship between ABO blood groups and the diarrheal disease caused by infection with Vibrio cholerae. Zhonghua Yu Fang Yi Xue Za Zhi, 1989 Mar, 23(2), 71 - 3 {The first isolation of Vibrio alginolyticus from samples which caused food poisoning}; Ji SP; We report an event of food poisoning traced to eating salted shrimps . Vibrio alginolyticus was shown to be the causative agent through epidemiological investigations and etiological tests . Vibro alginolyticus can bring about human septicaemia and wound infection and it was found in the feces of patients with diarrhea, but no determination on its pathogenicity was done . From the samples of food which led to food poisoning . Vibrio alginolyticus was isolated, and for the first time it was determined as a pathogen of food poisoning. Injury, 1989 Mar, 20(2), 87 - 91 Outboard motor propeller injuries; Kutarski PW; Seven cases of injury from an outboard motor propeller are reported and the literature reviewed . The injuries are uncommon, but appear to have a high rate of morbidity and mortality . Wound contamination at the time of injury is discussed . These wounds are best treated by excision of dead tissue and delayed primary suture plus antibiotics, which should include cover against the Vibrionaceae. Pediatr Emerg Care, 1989 Mar, 5(1), 27 - 8 Vibrio fluvialis, an unusual pediatric enteric pathogen; Bellet J et al.; An infant with diarrhea was discovered to have Vibrio fluvialis, an enteric pathogen not previously reported in children in the United States . This patient had an uncharacteristically mild clinical course and did not require hospitalization or antibiotics . Clinicians treating patients with diarrhea should include this organism in the differential diagnosis if their patients or patients' contacts have eaten seafood or been near bodies of seawater. J Bacteriol, 1989 Mar, 171(3), 1288 - 93 Identification of toxS, a regulatory gene whose product enhances toxR-mediated activation of the cholera toxin promoter; Miller VL et al.; We describe the cloning of the toxS gene from Vibrio cholerae E1 Tor strain E7946 . This gene lies downstream from the toxR gene, which encodes the transcriptional activator for the cholera toxin (ctx) operon in V . cholerae . We show that ToxS acts in conjunction with ToxR to activate expression of the ctx operon in Escherichia coli . The classical strain 569B, which is attenuated for virulance but which synthesizes high levels of cholera toxin in vitro, carries a deletion of 1.2 kilobase pairs of DNA, downstream from the toxR gene, which removes toxS . We present evidence that toxS is the downstream gene in an operon with toxR. J Diarrhoeal Dis Res, 1989 Mar-Jun, 7(1-2), 13 - 7 An investigation of a cholera epidemic in Butiama village of the Mara Region, Tanzania; Killewo JZ et al.; An outbreak of cholera between 19 March and 10 April 1986 in Butiama village of the Mara Region, Tanzania, was investigated in a case-control study, to try to find out the source of infection and its mode of transmission . Sixty-seven patients, including 11 deaths occurring before 29 March, were used to describe the epidemic, but 26 of the first patients were interviewed using a questionnaire, and each was compared with age and sex-matched healthy next-door neighbours . They were questioned about likely risk factors . Though the first two patients were recorded from 2 of the 4 zones of the village, from the second day onwards they came concurrently from all the 4 zones . Also, the number of patients from each zone did not vary significantly . The number of patients reached its peak on 22 March . Females were five times more infected than males, but the case fatality rate was similar . Vibrio cholerae was not isolated from water and fish-scale samples, but a history of handling and eating fish, and attendance at social gatherings were significantly associated with the transmission of cholera . The origin of the outbreak appeared to be either multifocal or a common source with concurrent multiple exists. Exp Hematol, 1989 Mar, 17(3), 273 - 7 Neuraminidase pretreatment of donor lymphocytes and graft-versus-host disease; Stacey NH et al.; Treatment of murine CBA splenocytes with Vibrio cholerae neuraminidase (VCN) prior to engraftment into cyclophosphamide (CY) immunosuppressed Balb/c x CBA mice reduced the incidence of acute lethal graft-versus-host disease (GvHD) and delayed mortality in the later phase of the disease . In the same model, pretreatment of donor splenocytes with VCN also reduced splenomegaly . Engraftment of F1 mice with CBA cells was clearly demonstrated at day 30 after infusion . Treatment of spleen cells with VCN did not compromise their ability to protect mice against irradiation-induced lethality . Furthermore, enzyme treatment was found to have no adverse effects on helper (TH-) and B-cell activity or on suppressor (TS-)cell function in adoptive transfer assays . Therefore, whereas the mechanism of the effect of the VCN preparation remains to be established, it is clear that the treatment provides protection against GvHD. Gene, 1989 Feb 20, 75(2), 289 - 96 A plasmid vector and quantitative techniques for the study of transcription termination in Escherichia coli using bacterial luciferase; Peabody DS et al.; We have developed a plasmid expression vector for the study of transcription terminators in Escherichia coli that utilizes the lux genes coding for the enzyme luciferase of the bioluminescent marine bacterium, Vibrio harveyi . The pBR322-derived plasmid, called pHV100, contains the E . coli lac promoter, the polylinker regions from the plasmid vector pUC18, and the V . harveyi lux genes . Insertion of transcription termination sites into the polylinker region results in decreased luciferase expression . Because the bioluminescence genes are not indigenous to E . coli, their expression can be studied in virtually any host strain without the complications of background activity . This facilitates sensitive measurements of terminator efficiency in hosts containing termination factor mutations . Bioluminescence can be easily monitored with high sensitivity, using a rapid photographic technique or a more quantitative photometric assay. Gene, 1989 Feb 20, 75(2), 253 - 9 Chromosomal transfer and in vivo cloning of genes in Vibrio cholerae using RP4::mini-Mu; Srivastava R et al.; The RP4::mini-Mu replicon has been used to transfer chromosomal genes by conjugation and to clone in vivo metabolic, toxin and flagellar genes of Vibrio cholerae . RP4::mini-Mu was introduced into several strains of V . cholerae and these strains were mated with V . cholerae or Escherichia coli K-12 recipients . R'-episomes carrying the respective genes were maintained in recA recipients and were detected by complementation of auxotrophic, nontoxinogenic and aflagellate mutations in V . cholerae. Eur J Biochem, 1989 Feb 15, 179(3), 651 - 7 Synthesis of glycoconjugates derived from various lipopolysaccharides of the Vibrionaceae family; Banoub JH et al.; Conjugation of simple ketoses (such as 3-deoxy-D-manno-2-octulosonic acid and N-acetylneuraminic acid) and of various O-specific polysaccharides (from Aeromonas hydrophila and Aeromonas salmonicida) to the bifunctional spacer 1,6-hexanediamine, was achieved by reductive amination . The saccharide--1-(6-amino)-hexane alkyamines obtained were converted into the corresponding isothiocyanate derivatives and coupled to the free epsilon-amino group of lysine residues of the protein carrier bovine serum albumin . In similar manner, the aldehyde group introduced by selective periodate oxidation into the partially O-deacylated lipopolysaccharide of Vibrio anguillarum was conjugated to 1,6-hexanediamine, converted into the corresponding isothiocyanate and covalently attached to bovine serum albumin. J Bacteriol, 1989 Feb, 171(2), 731 - 6 Iron regulation of swarmer cell differentiation of Vibrio parahaemolyticus; McCarter L et al.; Vibrio parahaemolyticus has two distinct cell types, the swimmer cell and the swarmer cell, adapted for locomotion in different circumstances . The swimmer cell, produced when the bacterium is grown in liquid media, is a short rod with a single sheathed polar flagellum . The swarmer cell, produced when V . parahaemolyticus is grown on solidified media, is greatly elongated and synthesizes, in addition to the polar flagellum, numerous unsheathed lateral flagella which are responsible for translocation over surfaces . We are interested in understanding how this bacterium differentiates in response to contact with surfaces and have determined in earlier work that the polar flagellum acts as a tactile sensor which controls transcription of genes (laf) encoding the swarmer cell phenotype . Surface recognition involves sensing of forces that obstruct movement of the polar flagellum . In this report we show that a second signal, iron limitation, is also required for swarmer cell differentiation . Production of lateral flagella occurred only when polar flagellar function was perturbed and iron-limiting growth conditions were imposed . The same conditions were required to induce light production in strains of V . parahaemolyticus in which a laf gene was transcriptionally fused to the lux operon encoding the enzymes for bioluminescence . The lafA gene encoding the lateral flagellin subunit was cloned and used in Northern (RNA) blot measurements . Examination of mRNA levels revealed that transcription of lafA is dependent on growth in iron-depleted media . The control of differentiation by multiple environmental stimuli is discussed. Infect Immun, 1989 Feb, 57(2), 495 - 501 Human immune response to Vibrio cholerae O1 whole cells and isolated outer membrane antigens; Richardson K et al.; The serum immunoglobulin G (IgG) and mucosal secretory IgA (SIgA) response of human volunteers challenged with Vibrio cholerae O1 was analyzed for reactivity to V . cholerae O1 antigens by the immunoblot technique . Components of both in vitro- and in vivo (rabbit ligated ileal loop)-grown V . cholerae O1 were separated by sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis . Postchallenge serum IgG reacted uniquely with 15 antigens and with greater intensity than did prechallenge serum with at least 16 antigens . Serum IgG and SIgA reacted with antigens present in preparations from the homologous challenge strain of V . cholerae as well as antigens from strains of heterologous biotype or serotype . These heterologous antigens may represent antigens responsible for protection to rechallenge with a heterologous strain of V . cholerae . All the antigens detected by postchallenge jejunal fluid SIgA had an apparent molecular size of less than 25 kilodaltons . Serum IgG and jejunal fluid SIgA also reacted with antigens unique to in vivo-grown cells and several antigens in outer membrane preparations, suggesting that studies of protective immunity and V . cholerae O1 pathogenesis should include examination of both in vitro- and in vivo-grown V . cholerae O1 cellular antigens. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1989 Feb, 22(1), 75 - 81 {Ferric ammonium citrate as virulence-enhancing agent in the toxicity test of Vibrio cholerae and the potency assay of cholera vaccine}; Tsai MY et al.; Using ICR strain mice as experimental animals, 0.25% (w/v) ferric ammonium citrate (FAC) was a suitable substitute for 5% (w/v) mucin as a virulence-enhancing (VE) agent in the toxicity test of Vibrio cholerae and the potency assay of cholera vaccine . There was no significant lethal toxicity difference between 0.25% FAC and 5% mucin as VE agents on Vibrio cholerae in mice . By using 0.25% FAC as a VE agent in estimating the potency of cholera vaccine, the relative potency (R.P.) of the tested cholera vaccine (Inaba) to the reference cholera vaccine (Inaba) was 0.896 +/- 0.208, the variation coefficient (V.C.) was 0.232, the correlation between the immune dose and the ratio of mouse death (r2) was 0.9932; for the cholera vaccine (Ogawa), R.P . was 1.373 +/- 0.366, V.C . was 0.266 and r2 was 0.8231 . There was no significant difference between 5% mucin and 0.25% FAC used as VE agents in potency assay of cholera vaccine. Zh Mikrobiol Epidemiol Immunobiol, 1989 Feb, (2), 41 - 5 {Use of the DNA-DNA hybridization method on filters for determining the toxic potency of atypical strains of Vibrio cholerae isolated from natural sources}; Demkin VV et al.; The methods of the radioimmunoassay and the blot hybridization of restricted fragments of chromosomal DNA have been used for the characterization of V . cholerae atypical strains isolated from the natural environment . For all strains under study, the radioimmunoassay has been found to yield the most sharply defined data characterizing their atoxigenicity . The absence of the structural genes of toxin in the chromosomes of these strains has been shown by the method of blot hybridization . Some methodological simplifications of blot hybridization, having no adverse effect on the sensitivity of this method, have been tested. Pediatr Infect Dis J, 1989 Feb, 8(2), 105 - 9 Effects of undernutrition on infection with Vibrio cholerae O1 and on response to oral cholera vaccine; Glass RI et al.; The association between undernutrition and the risk of colonization and disease with Vibrio cholerae O1, concentrations of salivary IgA and the serologic response to infection and to orally administered cholera B subunit were examined prospectively in a family study in Bangladesh . Children ages 1 to 8 years who were family contacts of patients hospitalized with culture-confirmed cholera were visited within 24 hours of the hospitalization and daily for 10 days, queried for the presence of diarrhea and cultured for V . cholerae O1 . On Day 1 each child was weighed and saliva was collected to measure total IgA . On Days 1 and 21 blood was taken to assess vibriocidal and antitoxin titers, and on Days 1 and 2 B subunit or placebo was given orally as part of a trial to look for a toxin-blocking effect . Of 412 children enrolled in the study 35% (143) became infected with V . cholerae O1 and 49% (70) of these developed diarrhea . Undernutrition, defined in a child as weight less than 70% of the Harvard reference weight-for-age, was not associated with colonization, disease or the duration or severity of cholera . Moreover well-nourished children did not differ from undernourished children in their concentrations of salivary total IgA, initial serum antitoxin or vibriocidal antibodies or in their serologic response to colonization, disease or B subunit . The immune system in its response to cholera appears to be quite resistant to nutritional insults . The good antitoxin response to B subunit among undernourished children is of particular importance in considering the use of future oral cholera vaccines in areas where such undernutrition is common. J Gen Microbiol, 1989 Feb, 135 ( Pt 2), 345 - 51 Purification and some properties of a novel L-2,4-diaminobutyric acid decarboxylase from Vibrio alginolyticus; Nakao H et al.; Previous investigations have shown that members of the genus Vibrio possess a novel enzyme activity decarboxylating L-2,4-diaminobutyric acid (DABA) to 1,3-diaminopropane (DAP) . In this paper we describe the purification, by about 3600-fold, of the enzyme from V . alginolyticus . The purified enzyme was apparently homogeneous, and had a specific activity of 4200 nmol DAP min-1 (mg protein)-1 . The enzyme protein has an Mr of 450,000 +/- 20,000 and is apparently comprised of four identical subunits (Mr 109,000 +/- 1,000) . Neither 2,3-diaminopropionic acid, ornithine, lysine nor arginine served as substrates . Some properties of the enzyme were determined . Cultivation of this bacterium in the presence of added DABA brought about increased production of norspermidine (NSPD), characteristically present in this species as well as DAP, suggesting that the enzyme may be functionally implicated in the formation of DAP, a biosynthetic precursor of NSPD. Exp Hematol, 1989 Feb, 17(2), 106 - 9 Effects of in vivo neuraminidase on the regulation of erythropoiesis . II: Modulation of erythroid colony formation by thymic regulatory cells; La Russa VF et al.; Effects of the enzyme vibrio-cholerae neuraminidase (VCN) on the marrow-derived erythropoietic progenitor CFU-E and thymic regulatory cells were examined in vitro 1 and 24 h after i.v . injection of the enzyme . An in vivo enzymatic modification of bone marrow and thymic helper regulatory cell function occurs within 1 h after i.v . injection of VCN and results in suppression of both CFU-E colony formation and thymic helper cell function . These inhibitory effects of neuraminidase, however, are no longer detectable by 24 h after injection . More importantly, these inhibitory effects can be reversed by adding thymocytes from control animals to cocultures of enzymatically modified marrow or thymic regulatory cells . These findings: 1) suggest that regulatory cells from the bone marrow and thymus may be enzymatically modified in vivo in a reversible manner, suggesting a noncytotoxic effect of the enzyme on accessory cells, and 2) confirm the importance of sialic acid for the helper function but not for the suppressor function of thymocytes and CFU-E colony formation in vitro. J Bacteriol, 1989 Feb, 171(2), 1199 - 202 Regulation of luminescence by cyclic AMP in cya-like and crp-like mutants of Vibrio fischeri; Dunlap PV; Mutants of Vibrio fischeri MJ-1 (wild type) apparently deficient in adenylate cyclase (cya-like) or cyclic AMP receptor protein (crp-like) were isolated and characterized . Compared with MJ-1, the mutants produced low levels of luminescence and luciferase . Addition of cyclic AMP restored wild-type levels of luminescence and luciferase in the cya-like mutant but not in the crp-like mutant . The results are consistent with the hypothesis that in V . fischeri cyclic AMP and cyclic AMP receptor protein are required for induction of the luminescence system. Kansenshogaku Zasshi, 1989 Feb, 63(2), 156 - 61 {Two case reports of septic shock due to Vibrio vulnificus with liver cirrhosis}; Tateyama M et al.; We have recently experienced a case of Vibrio vulnificus septicemia which occurred in a patient with hepatic cirrhosis, and as we were able to give early antibiotic treatment, the patient survived . We would like to report this case here together with another case experienced 2 years ago . Case 1 was a 58-year-old male who was attending our hospital as an outpatient for hepatic cirrhosis . At 5:30 pm on August 8, 1987, he consumed abalone and giant clam and at 9 pm complained of high fever with shaking chills . He was admitted to our department as an emergency case . Cefoperazone was administered resulting in a decline of fever on the following day . During the course of treatment he fell transiently into pre-DIC, but due mainly to the administration of antibiotics his condition was subsided . Case 2 was a 53-year-old male who was under medical care in our hospital for grave hepatic cirrhosis . On October 11, 1985, he consumed sushi and two days later suffered chills and pyrexia . A blood culture revealed Vibrio vulnificus . His condition improved transiently with administration of Cefazolin, but oliguria, hypotension and ascites occurred subsequently, and finally the patient died on the 22nd day. Kansenshogaku Zasshi, 1989 Feb, 63(2), 138 - 44 Effects of salinity on the survival of non-O1 Vibrio cholerae under environments of low temperature; Uchiyama H et al.; Under low temperature conditions, the effects of survival and growth of non-O1 Vibrio cholerae were studied in laboratory microcosms . As the environmental factors, various concentrations of NaCl solution and that of mono- and bivalent cation added nutrient of polypeptone were used under several temperatures . The results of these experiments suggested an extended survival of non-O1 V . cholerae at low temperature . When a range of NaCl concentration in microcosms water was between 0.4% and 2.0% with absent of nutrient, the survival of this organism was good under the temperature from 10 degrees C to 20 degrees C . With added nutrient, when about 10 mM bivalent cation and over 0.3% NaCl co-existed, this strain was able to grow at 10 degrees C, and showed good survival . This finding suggested the possibility of survival under 10 degrees C. Biochim Biophys Acta, 1989 Jan 23, 1007(1), 84 - 90 Expression of bacterial luciferase genes from Vibrio harveyi in Bacillus subtilis and in Escherichia coli; Karp M; For gene expression and cell physiology studies of Gram-positive Bacillus subtilis, novel shuttle vectors which cause the host organism to produce light have been constructed . These vectors carry genes encoding luciferase from Vibrio harveyi, a selectable kanamycin marker and an origin of replication for Gram-negative and -positive bacteria . The effect of DNA-insert size on light production in Escherichia coli and in B . subtilis was studied by also cloning into the shuttle vector a gene whose product participates in fatty acid metabolism . B . subtilis containing lux genes was found to differ from its Gram-negative counterpart in light emission characteristics . After addition of the substrate, light emission by B . subtilis was rapid but it decayed quickly, showing biphasic kinetics . In E . coli light emission is continuous, reflecting constant regeneration of substrates for the luciferase reaction . A promoter cloning vehicle, pCSS117, was constructed by inserting a transcriptional termination loop in the upstream sequences of luciferase genes . Using this vector, the mode of action of promoters of interest can be studied in E . coli and in B . subtilis. FEMS Microbiol Lett, 1989 Jan 15, 48(2), 241 - 5 Purification of a TDH-related hemolysin produced by a Kanagawa phenomenon-negative clinical isolate of Vibrio parahaemolyticus 06: K46; Honda T et al.; A hemolysin produced by a clinical isolate of Kanagawa phenomenon-negative Vibrio parahaemolyticus 06: K46 was purified by 55% ammonium sulfate fractionation and successive column chromatographies on DEAE-cellulose, hydroxyapatite, Sepharose 4B and Mono Q . The purified hemolysin was physicochemically and immunologically identical with the Vp-TRH (V . parahaemolyticus thermostable direct hemolysin related hemolysin) recently described in V . parahaemolyticus 03: K6 (Honda et al . Infect . Immun . 56: 961-965, 1988) . This indicates that V . parahaemolyticus of Kanagawa-negative clinical isolates possessing not only 03: K6 but also different serotypes such as 06: K46 produce Vp-TRH . Production of Vp-TRH by most clinical isolates of Kanagawa-negative V . parahaemolyticus was also demonstrated . These results suggest the importance of Vp-TRH among clinical isolates of Kanagawa-negative V . parahaemolyticus. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1989 Jan, 270(3), 361 - 72 Antigenic analysis of Vibrio cholerate O1 by crossed immunoelectrophoresis; Kabir S; Antigens from Vibrio cholerae O1 were analyzed by crossed immunoelectrophoresis (CIE) using sera from immunized rabbits . Thirty different anode-migrating antigens were detected in sonicated antigen preparations of V . cholerae . These antigens were numbered in order to establish a reference precipitation pattern . Antigen no . 30 was identified as the lipopolysaccharide (LPS) antigen, because it reacted with (i) periodic acid-Schiff (PAS) reagent and (ii) the affinity-purified anti-LPS antibodies . Treatment with proteinase K demonstrated that most of the precipitation lines were due to proteins, a part of which were localised at the cell surface . The major outer membrane protein was found to be closely associated with the precipitation line due to the LPS (antigen no . 30) . The antigenicity and immunogenicity of V . cholerae cells killed by different methods (merthiolate, heat, phenol, formalin) were examined . As determined by CIE, killing with merthiolate preserved most of the major components of V . cholerae . Heat, phenol and formalin altered the antigenic mosaic of V . cholerae . These results suggested that CIE can be used to analyze several aspects of V . cholerae antigens. J Endocrinol, 1989 Jan, 120(1), 135 - 42 Stress alters immune function and disease resistance in chinook salmon (Oncorhynchus tshawytscha); Maule AG et al.; We examined the effects of acute stress on the immune system and disease resistance of juvenile chinook salmon (Oncorhynchus tshawytscha) in laboratory and clinical trials . Immune function, as measured by the ability of lymphocytes from the anterior kidney to generate specific antibody-producing cells (APC) in vitro, was depressed 4 h after stress, when plasma cortisol levels were highest . At the same time, resistance to the fish pathogen, Vibrio anguillarum, was also depressed . Compared with controls, plasma cortisol and APC of stressed fish were unchanged after 24 h, and disease resistance was enhanced as evidenced by higher survival rate and longer mean time to death of mortalities . After 7 days, even though numbers of APC were depressed, plasma cortisol concentration and disease resistance did not differ from controls . This pattern was generally the same, independent of the type of stress applied: i.e . being held out of water in a dipnet for 30 s, manipulation during hatchery operations for 4 h, or transportation for 9 h . These and earlier findings suggest that similar endocrine-immune interactions operate in the mammalian and salmonid systems during acute stress. Mutat Res, 1989 Jan-Feb, 225(1-2), 43 - 7 An adaptive response of Vibrio cholerae strain OGAWA 154 to furazolidone; Bhattacharya R; Since furazolidone is an antimicrobial drug, any possibility of its evoking an adaptive response appears to be very important . This response was studied in Vibrio cholerae cells as a model system . In order to determine this response, a dose-response relation of these cells to furazolidone and the kinetics of inactivation of the drug were studied . The study of the adaptive response of these cells to furazolidone reveals that cells treated with a low concentration of furazolidone for a particular period were 100% more resistant to the lethal effects of a subsequent challenging dose than control cultures . Variation of the challenging dose level showed better survival of adapted cells than control cells . A time-dependent response study reveals a maximum response at 15-30 min, and a gradual fall thereafter. Infect Immun, 1989 Jan, 57(1), 158 - 63 Effects of divalent cations and saccharides on Vibrio metschnikovii cytolysin-induced hemolysis of rabbit erythrocytes; Miyake M et al.; Divalent cations and polysaccharides such as inulin and dextran reversibly inhibited hemolysis of rabbit erythrocytes caused by Vibrio metschnikovii cytolysin . On the basis of the 50% inhibitory doses, the cations were divided into two groups, group I (Cd2+, Cu2+, Ni2+, Sn2+, and Zn2+) and group II (Ba2+, Ca2+, Co2+, Mg2+, Mn2+, and Sr2+) . Neither divalent cations nor polysaccharides interfered with the binding of toxins to the erythrocyte membrane . Group I cations disturbed tetramer formation of cytolysin on the cytolysin-lysed erythrocyte membrane, although group II cations and dextran did not affect the process . Erythrocytes treated with cytolysin in the presence of group II cations or dextran lysed after transfer to toxin- and inhibitor (group II cations or dextran)-free buffer at both 37 and 4 degrees C . However, erythrocytes treated in the presence of group I cations lysed at 37 degrees C but not at 4 degrees C, indicating that group I cations block the temperature-dependent lesion (tetramer)-forming step subsequent to the binding of cytolysin to the erythrocytes . The cytolysin-treated erythrocytes swelled in a colloid osmotic manner, and the swelling was preceded by the binding and the lesion-forming steps . It is also suggested that the lysis of the erythrocytes proceeds in a temperature-independent manner and that the cytolysin does not bind to the erythrocytes at 4 degrees C . These findings suggest that the sequence of V . metschnikovii cytolysin-induced hemolysis is defined by three steps: (i) a temperature-dependent binding step, (ii) a temperature-dependent lesion-forming step, and (iii) a temperature-independent lysis step. Infect Immun, 1989 Jan, 57(1), 117 - 20 Immunogenicity of two formulations of oral cholera vaccine comprised of killed whole vibrios and the B subunit of cholera toxin; Migasena S et al.; Two formulations of oral cholera vaccine were evaluated for safety and immunogenicity in adult volunteers in Thailand, an area with sporadic cholera outbreaks . One formulation consisted of 2 x 10(11) killed vibrios and 5 mg of cholera toxin B subunit, as was previously evaluated in North American volunteers, and the other consisted of 1 x 10(11) killed vibrios and 1 mg of B subunit, as was recently evaluated in a field trial in Bangladesh . Three doses of each formulation were given with citrate-bicarbonate buffer . Neither formulation had adverse effects . The formulations stimulated similar serum immunoglobulin G (IgG) and IgA responses to Vibrio cholerae lipopolysaccharide and cholera toxin and intestinal secretory IgA responses to lipopolysaccharide and toxin after three doses . The formulation containing twice the quantity of killed vibrios stimulated better vibriocidal responses, especially to Ogawa serotype . A formulation of oral vaccine containing more killed vibrios than were included in the vaccine studied in the Bangladesh field trial may provide greater protection against cholera. J Virol, 1989 Jan, 63(1), 392 - 7 Abortive replication of choleraphage phi 149 in Vibrio cholerae biotype el tor; Chowdhury R et al.; Choleraphage phi 149 adsorbed irreversibly to Vibrio cholerae biotype el tor cells, and 50% of the injected phage DNA bound to the cell membrane . Although no infectious centers were produced at any time during infection, the host macromolecular syntheses were shut off and the host DNA underwent chloramphenicol-inhibitable degradation . Synthesis of monomeric phage DNA continued similar to that observed in the permissive host . However, the concatemeric DNA intermediates produced were unstable and could not be chased to mature phage DNA . Pulse-labeling of UV-irradiated infected cells at different times during infection allowed identification of phage-specific proteins made in this nonpermissive host . Although most of the early proteins were made, only some of the late proteins were transiently synthesized. Nippon Geka Hokan, 1989 Jan 1, 58(1), 155 - 61 A case report of acute obstructive suppurative cholangitis in a non-0-1 Vibrio cholerae biliary carrier; Nishikawa M et al.; A case report is presented of a 73-year-old male who was seen with fever, jaundice, abdominal pain and central nervous system depression . He failed to respond to intensive antibiotic therapy, and subsequently acute obstructive suppurative cholangitis fully developed . Upon laparotomy, the patient's gallbladder was found to be enlarged with the bile from the gallbladder and bile duct itself containing a high pus content . Its cultured organism revealed non-0-1 Vibrio cholerae . To our knowledge, no prior case of acute obstructive suppurative cholangitis in a non-0-1 Vibrio Cholerae biliary carrier has been reported in Japan. Med Microbiol Immunol (Berl), 1989, 178(5), 279 - 87 Extracellular and surface-bound biological activities of Vibrio fluvialis, Vibrio furnissii and related species; Myatt DC et al.; Twenty-seven Vibrio strains were assessed for virulence-associated biological activities, including iron chelation, hydrolases, haemolysis and haemagglutination . All strains hydrolysed DNA, chitin, gelatin and casein, produced siderophores, and lysed red blood cells . All V . fluvialis, V . cholerae and V . mimicus strains exhibited diverse lipolytic activity distinct from more discriminate lipolysis by V . furnissii . V . furnissii manifested fibrin and mucin hydrolysis but no phosphate or esculin hydrolysis, for which V . fluvialis varied . No strains hydrolysed urea, alginate or keratin . V . fluvialis, V . furnissii and V . mimicus strains failed to exhibit the mannose-sensitive haemagglutination typical of V . cholerae . Some activities may distinguish otherwise phenotypically similar species . Species tested commonly possessed biological activities that may contribute to virulence, although there was no apparent correlation with isolation from human sources. Med Microbiol Immunol (Berl), 1989, 178(5), 245 - 53 Production of monoclonal antibodies against thermostable direct hemolysin of Vibrio parahaemolyticus and application of the antibodies for enzyme-linked immunosorbent assay; Honda T et al.; A total nine hybridoma cell lines that produced monoclonal antibodies against thermostable direct hemolysin (Vp-TDH), a possible pathogenic toxin, of Kanagawa phenomenon-positive Vibrio parahaemolyticus was isolated and characterized . These monoclonal antibodies (mAbs) were divided into a minimum of five different specificity groups, including mAbs specific to Vp-TDH and common to Vp-TDH and Vp-TRH, a Vp-TDH-related hemolysin produced by Kanagawa phenomenon-negative V . parahaemolyticus . An enzyme-linked immunosorbent assay (ELISA) using mAb-1-D, a mAb specific for Vp-TDH, was developed for specific detection of Vp-TDH . On the other hand, the ELISA using mAb-9-D, and mAb common to both Vp-TDH and Vp-TRH, could be used for detection of both Vp-TDH and Vp-TRH . Thus, by combining these two ELISAs differential detection of Vp-TDH and Vp-TRH can be performed . Hence, the two ELISAs were applied for various strains of V . parahaemolyticus and it was found that most Kanagawa phenomenon-positive and -negative clinical isolates produced Vp-TDH and Vp-TRH, respectively, but all environmental strains, that were Kanagawa phenomenon-negative, produced neither toxin. Microbiol Immunol, 1989, 33(8), 641 - 8 Sugar composition of the polysaccharide portion of lipopolysaccharides isolated from non-O1 Vibrio cholerae O2 to O41, O44, and O68; Kondo S et al.; The sugar composition of the polysaccharide portion of lipopolysaccharides (LPS) was determined for 42 serovars of non-O1 Vibrio cholerae, i.e., from serogroups O2 to O41, O44, and O68 . On the basis of their compositional sugar pattern, they were classified into 24 chemotypes . 2-Keto-3-deoxyoctonate (KDO) was totally undetectable by the conventional color test (Weissbach's reaction) under the conventional hydrolysis conditions . Instead, a kind of KDO-like substance, which was positive in the reaction but not identical to KDO, was found in serogroup O19 . Fructose, a characteristic sugar constituent of O1 V . cholerae LPS, was found in 33 serogroups but was absent from nine serogroups, approximately 20% of the members of this group so far examined. Microbiol Immunol, 1989, 33(8), 609 - 18 Hemolytic Vibrio cholerae O1 that is sensitive to Mukerjee's cholera phage IV and the phage produced by the hemolytic vibrio lysogenized with the infection of Mukerjee's cholera phage IV; Iwanaga M et al.; Strains of hemolytic Vibrio cholerae O1 (El Tor vibrio) which are sensitive to Mukerjee's cholera phage group IV were isolated from cholera patients in North-East Thailand in 1986 . Plaques of the phage on these hemolytic V . cholerae O1 were usually translucent but almost transparent on some strains, just like the plaques on non-hemolytic V . cholerae O1 (classical vibrio) . These hemolytic V . cholerae O1 were lysogenized with the infection of cholera phage IV, and the lysogenized strains produced phage different from cholera phage IV . These hemolytic strains were classified into Cured type in prophage typing of V . cholerae O1, El Tor, because they were also lysogenized with Kappa phage and were hemolytic . When Cured-type V . cholerae O1, El Tor previously isolated in various countries were examined for the sensitivity to cholera phage IV, some of the isolates were sensitive. Dev Comp Immunol, 1989 Spring, 13(2), 123 - 32 Activation of trout macrophages and production of CRP after immunization with Vibrio anguillarum; Kodama H et al.; To examine the mechanism of the protection of rainbow trout (Salmo gairdneri) against Vibrio anguillarum in the early stage of immunization, the activation of macrophages and production of C-reactive protein (CRP) were investigated . Fish immunized with formalin-killed bacteria emulsified in Freund's complete adjuvant (FCA) resisted intraperitoneal challenge with living bacteria seven and ten days after immunization . The activation of macrophages was demonstrated by a significant increase of the chemiluminescent (CL) response and phagocytic activity . These fish also showed a significant increase of the CRP level in sera . Fish immunized with V . anguillarum alone or injected with FCA, however, did not resist the challenge . Though FCA itself increased CRP level and the sera enhanced phagocytic activity, increase of CL activity was weak . These results indicated that the increase of CL activity and opsonising effect of CRP on the phagocytosis of specifically activated macrophages concern to host defense in the early stage of infection. J Hyg Epidemiol Microbiol Immunol, 1989, 33(2), 219 - 28 Serovars of Vibrio parahaemolyticus; Aldova E; Seashore water samples collected along the coastline in Bulgaria and Rumania contained in large numbers OK serovars of V . parahaemolyticus; some of these had been isolated repeatedly over an extended time period: 01 K32, 03 K30, 03 K48, 04 K37, 04 K53, 05 K17, 05 K30 . The serovar 05 K17 was virtually present in all water samples and was also isolated from a case of purulent ear infection in a child from Burgas . In contrast, strains recovered from Asian and African coastal water had different K antigens and were never identified in Europe . Two strains of V . parahaemolyticus (serovars 05 K15 and 07 K10) had positive swarming growth resembling that of V . alginolyticus . The first of these was Kanagawa-positive and was isolated from a case of severe diarrhea in Brazzaville . Vibrio parahaemolyticus isolates came from marine or brackish water specimens collected on sand banks, 3 strains were recovered from marine or brackish water in Africa . Vibrio harveyi, a sucrose-negative species important from differential diagnostic aspects, has been isolated from seashore water samples collected on coarse-sand or pebbly beaches. Eksp Onkol, 1989, 11(3), 35 - 8 {Transplantability and immunogenicity in mice of lymphoma Nk/Ly cells modified by concanavalin A and neuraminidase}; Bratus' GG et al.; The modifying effect of con A, Vibrio cholerae and non-Vibrio cholerae neuraminidase (VCN, NVCN) on the proliferative potential and immunogenicity of experimental lymphoma NK/Ly cells is shown in vitro and in vivo . The efficacy of immunization by the modified tumour cells of mice with transplantable lymphoma is evaluated . A considerable decrease in the proliferative potential of transplantability and a moderate increase in immunogenicity of Con A treated cells were determined in vitro . The high immunogenicity of VCN-modified cells was found within the limits of the studied immunologic parameters . However, the VCN-MTC immunotherapy had no advantages over Con A-MTC immunotherapy . It is found that the modifying effect of NVCN on a tumour cell is inferior to the effect of Con A and VCN. J Pediatr Gastroenterol Nutr, 1989 Jan, 8(1), 81 - 4 Turning off the diarrhea: the role of food and ORS; Molla AM et al.; Ninety-three boys aged 5 years or less who had diarrhea due to Vibrio cholerae were randomly assigned to treatment with glucose oral rehydration salt (ORS) or rice-based ORS . For the first 24 h, ORS only was given to all the patients . During the next 24 h, ORS and normal food were given . The efficacy of the two types of ORS was compared in terms of ORS intake, stool output, change in hematocrit reading, serum specific gravity, and increase in body weight . At the end of the first 24 h of treatment, a 50% reduction in ORS intake and stool output was observed in the 47 patients randomly assigned to receive rice ORS as compared with the 46 patients who received glucose ORS . During the second 24 h of treatment, a significant reduction in the stool output was noticed in the glucose ORS group, making the efficacy of glucose ORS equal to that of rice ORS . The study suggests that normal food can impart some of the superiority of "super" ORS to standard glucose ORS with regard to reduction of stool volume. Microbiol Immunol, 1989, 33(12), 1045 - 52 Chemotaxonomic study of Vibrio cholerae bio-serogroup Hakata on the basis of the sugar composition of the polysaccharide portion of its lipopolysaccharides; Kondo S et al.; A chemotaxonomic study was carried out on 31 strains of non-O1 Vibrio cholerae bio-serogroup Hakata, isolated in Japan, which possesses the Inaba antigen C of O1 V . cholerae . On the basis of the compositional sugar pattern of the polysaccharide portion of their lipopolysaccharides, the 23 strains isolated from the environment were separated into two groups, one (20 strains) containing mannose, glucose, fructose, L-glycero-D-mannoheptose, glucosamine, perosamine, quinovosamine, and an unidentified amino sugar AS, and the other (3 strains) containing two additional sugars, galactose and a trace amount of galactosamine . All of the eight strains isolated from imported seafoods belonged to the former group. Scand J Infect Dis, 1989, 21(6), 727 - 31 Vibrio vulnificus septicaemia presenting as spontaneous necrotising cellulitis in a woman with hepatic cirrhosis; Arnold M et al.; Vibrio vulnificus is a virulent marine organism commonly found in Hong Kong coastal waters which contaminates local sea-food . It may produce a primary septicaemia, often associated with secondary skin lesions, following ingestion of raw shell fish . We report a rapidly fatal case of primary V . vulnificus septicaemia in a 50-year-old housewife with post-hepatitic cirrhosis presenting as spontaneous necrotising cellulitis of the legs . V . vulnificus infection should be considered in patients with a history of liver disease with acute septicaemia and characteristic skin lesions. Scand J Infect Dis, 1989, 21(6), 721 - 6 Vibrio vulnificus infection; Chuang YC et al.; We report 3 cases of Vibrio vulnificus infections from Taiwan . Patient 1, who manifested symptoms of primary septicemia, died after 2 days . Patient 2, who had a wound infection and signs and symptoms of sepsis but negative blood cultures, responded to tobramycin and chloramphenicol plus surgical debridement, and recovered after 26 days of hospitalization . Patient 3 had secondary septicemia originating from a wound inflicted by a shrimp . Originally, the patient seemed to respond to ceftazidime and amikacin treatment along with surgical debridement, but subsequently died from adult respiratory distress syndrome (ARDS) induced by several episodes of aspiration which occurred after initial clinical improvements . We conclude that, for patients with severe wounds and evidence of V . vulnificus infection, an appropriate, powerful antibiotic, such as one of the third generation cephalosporins should be used as initial therapy unless the nature of the infection indicates other treatment. J Gen Microbiol, 1989 Jan, 135 ( Pt 1), 111 - 20 Expression and detection of different biotype-associated cell-bound haemagglutinins of Vibrio cholerae O1; Jonson G et al.; The expression of two cell-bound haemagglutinins, one sensitive to L-fucose (FSHA) and the other to D-mannose (MSHA), on Vibrio cholerae O1 strains of both the classical and the El Tor biotypes was studied by (i) agglutination of chicken and human group O erythrocytes in the presence of L-fucose or D-mannose, (ii) binding of the bacteria to L-fucose- and D-mannose-coated agarose beads, and (iii) agglutination of the bacteria by 'biotype-specific' antisera . All of the 12 classical strains studied that were isolated before 1979 gave FSHA of human O erythrocytes whereas only 6 of 17 classical strains isolated during recent epidemics expressed FSHA; a few of the classical strains expressed MSHA in addition to FSHA . All the El Tor strains gave MSHA of chicken erythrocytes and one strain also expressed FSHA . Both the cell-bound HAs were optimally expressed during the exponential phase of growth; FSHA markedly decreased during the late exponential phase while the MSHA usually persisted into the stationary phase . The expression of FSHA and MSHA correlated very well with the direct binding of vibrios to fucose- and mannose-coated agarose beads, respectively . Antiserum 'specific' for classical vibrios agglutinated classical strains expressing FSHA and also the El Tor strain exhibiting FSHA . Similarly, the anti-El Tor serum agglutinated all El Tor strains and also classical strains expressing MSHA, suggesting that the 'biotype-specific' sera were specific for the biotype-associated cell-bound HAs. Toxicon, 1989, 27(4), 439 - 64 Detection of Vibrio vulnificus cytolysin in V . vulnificus-infected mice; Gray LD et al.; Enzyme-linked immunosorbent assays using polyclonal and monoclonal antibodies specific for V . vulnificus cytolysin detected the toxin in an extract of skin lesions and in serum from mice showing local and systemic V . vulnificus disease after subcutaneous injection of the bacterium . The cytolysin also was detected in skin lesions by an indirect immunofluorescence procedure using polyclonal and monoclonal antibodies . Our findings provide direct evidence that the cytolysin is produced in vivo during the development of the disease process, and this observation is consistent with the hypothesis that the toxin is involved in the pathogenesis of V . vulnificus disease. Gastroenterol Clin Biol, 1989 Jan, 13(1), 18 - 24 {Adsorption potency of 2 clays, smectite and kaolin on bacterial enterotoxins . In vitro study in cell culture and in the intestine of newborn mice}; Brouillard MY et al.; The use of clays in the treatment of enterocolitis is justified by their ability to adsorb viruses, biliary acids and bacterial toxins secreted into the intestinal lumen . We have studied the in vitro inactivation of the LT toxins of Vibrio cholerae and E . coli, the ST toxin of ETEC and the verotoxin of EHEC . These various toxins were incubated with two types of clays, smectite and kaolin, to investigate the influence of dose, pH variations and the duration of contact of the clays with the toxins . Irrespective of their presence or absence in the supernatant, the biological activity of the toxins was assessed in cell culture and in the newborn mouse test . Both clays inactivated the LT toxin . Smectite was more efficient than kaolin as it was active immediately especially at the pH of intestinal chyme . The LT toxins were adsorbed on the clays by hydrogen bonding . This permitted the segregation of the toxins and prevented them from being fixed to the membrane receptors on the cells . The two clays were ineffective against the verotoxin of EHEC when the pH was alkaline although they were more efficient at acid pH . ST toxin of ETEC was slightly adsorbed by smectite and kaolin. Mutat Res, 1989 Jan, 210(1), 149 - 56 Lack of umuDC gene functions in Vibrio cholerae cells; Ghosh SK et al.; Attempts to identify an umuDC analog, using interspecific complementation of Escherichia coli mutants with plasmids containing a gene bank of Vibrio cholerae, were not successful . The DNA from none of the vibrio species examined including marine vibrios hybridized to E . coli umuC and umuD gene sequences . These cells are not mutable by ultraviolet (UV) light and cannot Weigle-reactivate UV-irradiated choleraphages, suggesting that vibrios are deficient in the umuDC operon . This possibility is supported by the fact that when the plasmid pKM101 carrying the mucAB genes is introduced into V . cholerae cells, they acquire the UV-mutable phenotype and UV-irradiated choleraphages can be Weigle-reactivated. J Surg Oncol, 1989 Jan, 40(1), 34 - 7 Controlled clinical trial of adjuvant immunotherapy with BCG and neuraminidase-treated autologous tumour cells in large bowel cancer; Gray BN et al.; A controlled, randomised clinical trial of immunotherapy was performed in 301 patients with stage B or C colorectal cancer . The immunotherapy treatment consisted of 18 vaccinations over a 2 year period following surgery with a combination of BCG given by scarification plus subcutaneous injection of Vibrio cholera neuraminidase (VCN)-modified autologous tumour cells . Five year follow-up has now been completed in all patients . The immunotherapy did not alter either the disease-free interval or the overall survival of patients in comparison with a control group of patients not receiving immunotherapy. Arch Microbiol, 1989, 152(6), 542 - 9 Nucleotide sequence of the Vibrio alginolyticus glnA region; Maharaj R et al.; The nucleotide sequence of a 4 kb fragment containing the Vibrio alginolyticus glnA, ntrB and ntrC genes was determined . The upstream region of the glnA gene contained tandem promoters . The upstream promoter resembled the consensus sequence for Escherichia coli sigma 70 promoters whereas the presumptive downstream promoter showed homology with nitrogen regulated promoters . Four putative NRI binding sites were located between the tandem promoters . The ntrB gene was preceded by a single presumptive NRI binding site . The ntrC gene was located 45 base pairs downstream from the ntrB gene . The V . alginolyticus ntrB and ntrC genes were able to complement ntrB, ntrC deletions in E . coli. Microbiol Immunol, 1989, 33(1), 1 - 9 Pili of Vibrio cholerae O1 biotype E1 Tor: a comparative study on adhesive and non-adhesive strains; Iwanaga M et al.; Pili were found on the cell surface of non-adhesive Vibrio cholerae O1 Biotype E1 Tor as well as the adhesive strain . Purified pili of the adhesive and non-adhesive strains were morphologically, electrophoretically, and immunologically, indistinguishable from each other . The molecular weights of both pilin (subunit protein of the pilus) were about 16,000 daltons as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . These 16 kDa pili are different from the pilus colonization factor, which is a 20.5 kDa protein, reported by Taylor et al . The 16 kDa pili of Vibrio cholerae O1 Biotype E1 Tor have hemagglutinating activity, but may have no role in colonization, because non-adhesive strains also have such pili. Arch Microbiol, 1989, 151(5), 439 - 44 A primary respiratory Na+ pump of an anaerobic bacterium: the Na+-dependent NADH:quinone oxidoreductase of Klebsiella pneumoniae; Dimroth P et al.; Membranes of Klebsiella pneumoniae, grown anaerobically on citrate, contain a NADH oxidase activity that is activated specifically by Na+ or Li+ ions and effectively inhibited by 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO) . Cytochromes b and d were present in the membranes, and the steady state reduction level of cytochrome b increased on NaCl addition . Inverted bacterial membrane vesicles accumulated Na+ ions upon NADH oxidation . Na+ uptake was completely inhibited by monensin and by HQNO and slightly stimulated by carbonylcyanide-p-trifluoromethoxy phenylhydrazone (FCCP), thus indicating the operation of a primary Na+ pump . A Triton extract of the bacterial membranes did not catalyze NADH oxidation by O2, but by ferricyanide or menadione in a Na+-independent manner . The Na+-dependent NADH oxidation by O2 was restored by adding ubiquinone-1 in micromolar concentrations . After inhibition of the terminal oxidase with KCN, ubiquinol was formed from ubiquinone-1 and NADH . The reaction was stimulated about 6-fold by 10 mM NaCl and was severely inhibited by low amounts of HQNO . Superoxide radicals were formed during electron transfer from NADH to ubiquinone-1 . These radicals disappeared by adding NaCl, but not with NaCl and HQNO . It is suggested that the superoxide radicals arise from semiquinone radicals which are formed by one electron reduction of quinone in a Na+-independent reaction sequence and then dismutase in a Na+ and HQNO sensitive reaction to quinone and quinol . The mechanism of the respiratory Na+ pump of K . pneumoniae appears to be quite similar to that of Vibrio alginolyticus. Microbiol Immunol, 1989, 33(1), 11 - 21 Characterization of putrescine production in nongrowing Vibrio parahaemolyticus cells in response to external osmolality; Yamamoto S et al.; Nongrowing Vibrio parahaemolyticus cells rapidly produced putrescine (Put) from added arginine when subjected to a low osmotic stress . This phenomenon was characterized in connection with a regulatory mechanism of the responsible enzymes, arginine decarboxylase (ADC) and agmatine ureohydrolase (AUH) . NaCl, KCl, LiCl, sucrose, and glycerol were used as solutes to prepare the resuspending media with various osmolalities . Regardless of whether the solutes were electrolytes or non-electrolytes, exposure of cells to low osmolality brought about instantaneous increases in both intra- and extracellular Put contents without significant changes in the contents of other polyamines . This acceleration in Put production was accompanied by no increases in the specific activities of ADC and AUH . On the other hand, when cells were exposed to the osmolality equivalent to 2 or 5% NaCl, all solutes except for glycerol did not cause a remarkable variation in the intracellular Put content, while the amount of Put in the medium varied depending on the solute used; sucrose and glycerol still greatly prompted Put production, as judged by high Put contents in the media, even at the osmolality equivalent to 5% NaCl . The cation efflux from cells, measured as the K+ release, was observed whenever the increase in Put production occurred . Furthermore, in vitro experiments showed that NaCl and KCl inhibited ADC to a similar extent, about 70% inhibition being observed at 200 mM . However, AUH was not affected by these compounds . These results suggest that the reduction in the concentrations of Na+ and K+ predominantly present in cells may cause the increase in activity of the preexisting ADC, which leads to the enhancement of Put production. Chin J Biotechnol, 1989, 5(1), 19 - 26 Preparation of high purity IgG and its application in screening for clones positive for Vibrio succinogenes L-asparaginase gene; Guan YQ et al.; A previous paper reported application of L-asparaginase IgG from Vibrio succinogenes in screening for positive clones . This paper describes the detailed procedure for preparation of high-purity IgG, an essential step of which involves coupling of a cell-free extract of host E . coli Y 1090 to Sepharose 4B beads and adsorption of the non-specific antigen components by passage of the polyvalent IgG through this affinity column . The IgG thus obtained exhibited improved specificity and was utilized as a radioimmunologic and horseradish peroxidase-linked immunologic probe in screening for positive clones . These IgG probes had the advantages of low background, high sensitivity, and good reliability. J Med Microbiol, 1989 Jan, 28(1), 33 - 7 Immunobiological relationships among new cholera toxins produced by CT gene-negative strains of Vibrio cholerae O1; Saha S et al.; The enterotoxic activities of partially purified new cholera toxins (NCT) prepared from the CT gene-negative strains of Vibrio cholerae serotype O1 isolated from cases of diarrhoea in man and from diverse environmental sources were assayed in the rabbit ileal-loop model and the toxin unit was calculated in micrograms of protein content . The secretory activities of one unit of homologous and heterologous enterotoxins were completely neutralised by 2.5 units of crude antiserum raised against one NCT preparation . In immunodiffusion tests, the NCT preparations from all strains tested gave precipitin bands against one antiserum showing reaction of complete identity . Thus, the present study clearly demonstrated that NCTs elaborated by CT gene-negative strains of V . cholerae O1 are immunobiologically similar. Proc Natl Acad Sci U S A, 1989 Jan, 86(2), 481 - 5 Recombinant system for overexpression of cholera toxin B subunit in Vibrio cholerae as a basis for vaccine development; Sanchez J et al.; We have constructed an overexpression system in which the gene encoding the B subunit of cholera toxin (CTB) was placed under the control of the strong tacP promoter in a wide host range plasmid . Recombinant nontoxigenic classical and E1 Tor Vibrio cholerae strains of different serotypes harboring this plasmid excreted 10- to 100-fold higher amounts of CTB than any other wild-type or recombinant strain tested and may therefore be useful killed oral vaccine strains . The manipulations to place the CTB gene under tacP also included, by design, the introduction of single enzyme restriction sites for gene fusions to the CTB amino terminus . Cloning into these sites allows construction of CTB-derived hybrid proteins carrying various putative vaccine peptide antigens. Microbiol Immunol, 1989, 33(10), 833 - 41 Sugar composition of the polysaccharide portion of lipopolysaccharides of Vibrio fluvialis, Vibrio vulnificus, and Vibrio mimicus; Iguchi T et al.; A chemotaxonomic study was carried out on Vibrio fluvialis and V . vulnificus on the basis of the sugar composition of the polysaccharide portion of their lipopolysaccharides (LPS) . A previously developed rapid method of preparing samples for compositional sugar analysis was employed . Nineteen O-serogroups of V . fluvialis were divided into 14 chemotypes while seven O-serogroups of V . vulnifucus were divided also into seven chemotypes since the polysaccharide portion of LPS of each serogroup has a different sugar composition from that of the other serogroups . Close similarities in the sugar composition of the same portion were demonstrated between serologically cross-reacting non-O1 group V . cholerae and V . fluvialis, and non-O1 V . cholerae and V . mimicus. Lab Delo, 1989, (6), 63 - 5 {A method of fixation and disinfection of microbial cells on magnetic sorbents}; Podzolkova GG et al.; The 96% ethanol has been found fairly effective for the fixation and disinfection of Yersinia pestis, Vibrio cholerae, Pseudomonas mallei, and Pseudomonas pseudomallei after their contact with magnetic polyacrylamide sorbents containing specific immunoglobulins for ligands . Such step is recommended for the quantitative immunofluorescence test employed for rapid analysis, diagnosis, and identification of the above microorganisms. Lab Delo, 1989, (