Vibriosis

Zh Mikrobiol Epidemiol Immunobiol, 1987 Nov, (11), 41 - 4
{Systemic immunity of experimental animals immunized with cholera vaccines}; Genova IuG et al.; The levels of antitoxic and vibriocidal antibodies in the sera of suckling rabbits after their parenteral immunization with cholera vaccine, cholera toxoid and a combination of cholera vaccine and toxoid were examined . Cholera vaccine induces intensive production of vibriocidal antibodies, and cholera toxoid, of antitoxic antibodies . The parenteral administration of the serum of rabbits immunized with cholera toxoid neutralized the action of cholera toxin in the small intestine of suckling rabbits . The complex preparation combines the properties of the corpuscular vaccine and the toxoid, inducing the production of both vibriocidal and antitoxic antibodies.

Appl Environ Microbiol, 1987 Nov, 53(11), 2696 - 8
Hemolytic activity of and lethal toxin production by environmental strains of Vibrio parahaemolyticus; Sarkar BL et al.; Repeated subculturing of Kanagawa-negative strains of Vibrio parahaemolyticus on Wagatsuma agar induced the production of a hemolysin which was not the thermostable direct hemolysin . Crude hemolysin exhibited a 30 to 40% lethal toxicity in mice after intraperitoneal injection . A 21-kilodalton protein band was observed with all the environmental isolates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Results suggested that a certain percentage of environmental strains of V . parahaemolyticus is responsible for pathogenesis.

Zh Mikrobiol Epidemiol Immunobiol, 1987 Nov, (11), 60 - 3
{Potential use of serological methods in detecting cholera toxin}; Emdina IA et al.; In the study of 50 Vibrio cholerae museum strains, 45 of them producing cholerigenic effect in suckling rabbits, cholera toxin, determined by means of the passive immune hemolysis (PIH) test, has been detected in the supernatant of the culture fluid of only two strains: V . cholerae 569 B, a well-known producer of cholera toxin, and V . cholerae (eltor) 1310, from whose population a toxigenic variant has been obtained by selection . To study the capacity of V . cholerae for producing toxin in vitro, in six cholerigenic strains, besides the supernatant of their culture fluids, also protein fractions, cell lysates and membrane fractions have been studied in the PIH test . In all these strains cholera toxin has been detected only in membrane fractions, which should be taken into consideration in the serological evaluation of the toxigenicity of V . cholerae.

J Gen Microbiol, 1987 Nov, 133 ( Pt 11), 3265 - 70
Reversal by cyclic AMP of the urea-induced inhibition of synthesis of a catabolite-repressible enzyme in Vibrio cholerae; Chakravarti D et al.; Low concentrations of urea, which did not inhibit the synthesis of the catabolite nonrepressible enzyme alkaline phosphatase in Vibrio cholerae, or markedly affect its overall growth, specifically inhibited the expression of the tryptophanase operon in a temperature-dependent manner . However, in contrast to what is found in Escherichia coli, this urea-induced inhibition of tryptophanase synthesis in V . cholerae could be almost completely relieved by exogenously added cyclic AMP . The possible mechanism of the process is discussed.

Proc Soc Exp Biol Med, 1987 Nov, 186(2), 174 - 82
Synthesis of protein in intestinal cells exposed to cholera toxin; Peterson JW et al.; The mechanism by which cyclic adenosine monophosphate (AMP), formed by intestinal epithelial cells in response to cholera toxin, ultimately results in alterations in water and electrolyte transport is poorly understood . Several studies have indicated that inhibitors of transcription or translation block much of the transport of ions and water in the intestine and edema formation in tissue elicited by cholera toxin . Data presented in this study confirmed the inhibitory effects of cycloheximide on cholera toxin-induced fluid accumulation in the rabbit intestinal loop model . Neither cycloheximide nor actinomycin D altered the amount of cyclic AMP that accumulated in intestinal cells and Chinese hamster ovary cells exposed to cholera toxin . An increase in {3H}leucine incorporation was readily demonstrable in intestinal epithelial cells from rabbits challenged with Vibrio cholerae . Similarly, intestinal epithelial cells incubated with cholera toxin for 4 hr synthesized substantially more protein than controls as determined by relative incorporation of {35S}methionine . Most of the new protein synthesized in response to cholera toxin was membrane associated and of high molecular weight . The possible significance of the toxin-induced protein relative to cholera pathogenesis was discussed.

Microb Pathog, 1987 Nov, 3(5), 365 - 75
Immunoglobulins in bile and serum of the rabbit associated with protection after Vibrio cholerae infection and vaccination; Rijpkema SG et al.; Previous studies have shown that cholera, as well as protective immunity against infection with Vibrio cholerae, can be induced in the rabbit . This protection is long-lasting (up to 30 months) and is characterized on challenge by rapid, symptom-free disappearance of V . cholerae from the intestine; we therefore believe this to be vibriocidal protection . In this study, we analysed the humoral and secretory immune response against various subcellular V . cholerae components in vibriocidally protected, non-vibriocidally protected, and unprotected animals . Only vibriocidal protection was found to be associated with high levels of biliary IgA directed against lipopolysaccharide O antigen . We did not find such a correlation between either type of protection and response in serum . Therefore, anti-lipopolysaccharide antibodies are essential in protection against experimental infection with V . cholerae.

J Wildl Dis, 1987 Oct, 23(4), 666 - 8
Vibrio damsela infection in a stranded leatherback turtle (Dermochelys coriacea); Obendorf DL et al.; Necropsy of a stranded adult leatherback turtle (Dermochelys coriacea) determined that the animal died as a result of valvular endocarditis and septicemia . Vibrio damsela was isolated from the endocardial thrombus . The route of entry for infection probably was through the gastrointestinal tract.

J Gen Microbiol, 1987 Oct, 133 ( Pt 10), 2859 - 64
Mechanism of haemolysis by Vibrio vulnificus haemolysin; Yamanaka H et al.; The haemolytic action of Vibrio vulnificus haemolysin (VVH) was compared to that of streptolysin O (SLO) . Both were cholesterol-binding haemolysins, but differed in the release of haemoglobin (Hb) . In the first step of haemolysis, the haemolysins were temperature-independently bound to the cholesterol site on the target erythrocyte membrane . This was followed by the rapid release of K+, which is an intra-erythrocyte marker . Hb was then released, in different ways . In the case of VVH, Hb was released slowly after a relatively long lag, whereas with SLO, Hb was released as rapidly as K+ . Haemolysis by VVH was inhibited by the addition of 30 mM-dextran 4 (mean Mr 4000), which is considered to be an effective colloid-osmotic protectant . The results therefore indicated that haemolysis by VVH (like that by Escherichia coli alpha-haemolysin and Staphylococcus aureus alpha-toxin) was caused by a colloid-osmotic mechanism . Both K+ and Hb release caused by VVH proceeded temperature-dependently, and the membrane fluidity of liposomes prepared with lipids extracted from sheep red blood cell membranes increased above 20 degrees C . These results suggest that the temperature-dependence of the haemolysis by VVH is due to the requirement for an increase in the membrane fluidity during the formation of a transmembrane pore.

J Gen Microbiol, 1987 Oct, 133 ( Pt 10), 2853 - 7
Variability of haemolysin(s) produced by Vibrio vulnificus; Okada K et al.; The peptide composition and antigenic cross-reactivity of partially purified and concentrated haemolysins of 16 strains of Vibrio vulnificus were examined by SDS-PAGE and immunoblotting analysis, using a monoclonal antibody (MAb), 6F8D, raised against the haemolysin . All strains produced a common peptide of 36 kDa and the MAb reacted with this peptide . In some strains, larger molecules, including a 56 kDa peptide, were produced, but the MAb did not react with this peptide . The haemolytic activity of the strains was effectively neutralized by the MAb, except in the case of strains producing the 56 kDa peptide . These findings indicate that the 36 kDa haemolysin is common to all 16 strains and that V . vulnificus can produce a second haemolysin which differs in molecular mass and antigenicity.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1987 Oct, 266(3-4), 552 - 62
Protective immunity against Vibrio cholerae infection in the rabbit; Guinee PA et al.; The DIC model (Duodenal Inoculation with ligation of the Cecum in rabbits) was employed to study experimentally induced cholera and the related protective immunity . Duodenal inoculation (DI) without ligation of the cecum with live V . cholerae organisms did not cause any disease symptom but induced protection against subsequent challenges with homologous and heterologous organisms for up to 24 months . After 30 months this protective immunity began to decrease . A similar protective immunity could be induced by administration of the A- B+ derivative CVD101 of V . cholerae strain 395 . This type of experiment can only be done successfully with conventional, healthy rabbits held under low stress conditions . A so-called specific pathogen-free rabbit breed was found to be entirely unsuitable . Duodenal inoculation with heat- or merthiolate-inactivated V . cholerae for a prolonged period of time by means of an intestinal osmotic minipump did not induce protection . Injection of heat-inactivated V . cholerae material into the Peyer's patches sometimes led to protection, suggesting that a thermostable antigen, possibly lipopolysaccharide, is one of the major protective antigens . Duodenal administration of a combination of inactivated V . cholerae serotypes Ogawa and Inaba cells and 1 mg B subunit of the V . cholerae enterotoxin by up to three inoculations protected only 3 out of 12 rabbits against challenge . The results obtained on the rabbit model are discussed in relation to the efficacy of this vaccine in human volunteers and in a recent field test.

J Gen Microbiol, 1987 Oct, 133 ( Pt 10), 2751 - 7
Properties of the membrane-bound 5'-nucleotidase and utilization of extracellular ATP in Vibrio parahaemolyticus; Sakai Y et al.; Vibrio parahaemolyticus utilized ATP, ADP or AMP as the sole source of carbon . About three times higher activity of membrane-bound 5'-nucleotidase was observed in cells grown in the presence of these nucleotides than in their absence: and therefore the enzyme seems to be inducible . Since the 5'-nucleotidase activity could be measured with whole cells, the active site of this enzyme appears to be outwardly oriented . Both Mg2+ and Cl- were required for activity . Among the divalent cations tested, Mn2+ and Co2+ could replace Mg2+ to some extent, whereas Zn2+ strongly inhibited activity . Among the anions tested, Br-, I- and NO3- could replace Cl-, but SO4(2-) and CH3COO- could not . When cells were grown with ATP, Cl- was indispensable and Zn2+ strongly inhibited growth . Therefore, it is concluded that extracellular ATP and other 5'-nucleotides are cleaved by the membrane-bound 5'-nucleotidase outside the cells and that the adenosine produced is then utilized.

Infect Immun, 1987 Sep, 55(9), 2093 - 102
In vivo adherence and colonization of Vibrio cholerae strains that differ in hemagglutinating activity and motility; Teppema JS et al.; A scanning electron microscopic study was carried out to compare the in vivo pathogenicity of two strains of Vibrio cholerae in an adult rabbit ligated-gut test model . V . cholerae C5 (serotype Ogawa, biotype El Tor), a motile strain possessing hemagglutinating activity in vitro, and C21 (serotype Ogawa, classical biotype), a nonmotile strain possessing no hemagglutinating activity, were tested . Tissue samples from small intestinal loops were examined 3, 6, 9, and 12 h postinoculation . Contradictory to most published data, neither hemagglutinating activity nor motility appeared to be essential prerequisites for the pathogenesis of cholera in the experimental animal model used: nonmotile hemagglutinin-negative strain C21 adhered to and colonized the small intestine at least to the same extent as did motile hemagglutinin-positive strain C5 . Maximum colonization was seen at 9 h postinoculation for both strains . C5 and C21 vibrios caused comparable damage to the villi of the small intestine . The villous epithelium showed only mild changes during the first 9 h postinoculation . However, after 12 h the epithelium was seriously damaged concomitant with a decrease in the number of vibrios . Many villi showed partial or total denudation, owing to repelled epithelium, leaving a bare basal lamina with only some to moderate numbers of vibrios attached . Since similar changes were induced by pure cholera enterotoxin, these changes were likely the result of excessive fluid accumulation . From this study it is concluded that, at least in the animal model used, factors other than hemagglutinating activity and motility may also play a role in the association of V . cholerae with the small intestinal surface.

J Appl Bacteriol, 1987 Sep, 63(3), 255 - 60
Quantitative measurement of endotoxin in rainbow trout (Salmo gairdneri) serum by the chromogenic substrate method; Kodama H et al.; The amount of endotoxin in serum collected from normal rainbow trout (Salmo gairdneri) and trout inoculated with viable Vibrio anguillarum or lipopolysaccharide (LPS) extracted from bacteria was determined by the chromogenic substrate method . The mean values of endotoxin in four different groups of normal rainbow trout sera ranged from 31.9 to 65.3 pg/ml . When fish were inoculated with viable bacteria (1 x 10(8}, they became septicaemic and a large amount of endotoxin ng/ml) was detected in the sera . In fish inoculated with a smaller number of bacteria the amount of endotoxin was several times higher than that of normal fish in spite of failure of bacterial isolation . Although the endotoxin level in serum increased rapidly (greater than 100 ng/ml) after intraperitoneal inoculation with purified V . anguillarum LPS (540 micrograms), no fish died during the experiment . The high level of endotoxin in normal rainbow trout and the resistance of trout to endotoxin are in striking contrast to those of mammalian and avian species.

Infect Dis Clin North Am, 1987 Sep, 1(3), 687 - 707
Localized and systemic infection due to Vibrio species; Hill MK et al.; Many important species have been added to the genus Vibrio in the past several years . Vibrios have been associated with a wide variety of clinical syndromes ranging from mild gastroenteritis to life-threatening cellulitis . Most Vibrio infections follow consumption of raw shellfish or exposure to sea water . Although much has been learned about these organisms in the past several years, additional information is needed concerning pathogenesis, diagnosis, treatment, and prevention of Vibrio infections.

Mikrobiologiia, 1987 Sep-Oct, 56(5), 860 - 4
{Importance of Bdellovibrio in regulating microbial cenoses and self-purification processes in domestic sewage}; Lambina VA et al.; The bacterial parasite Bdellovibrio was directly proved to be involved in the regulation of microbial cenoses and in the self-purification of domestic waste waters . The incidence of heterotrophs, Gram-negative bacteria, E . coli and Bdellovibrio was followed up in dynamics in the microecological system of waste waters for ten days . In control experiments, bdellovibrions were removed using pteridine as a vibriostatic agent . In the absence of bdellovibrions, the cell number of the studied microorganisms did not increase after reaching a stationary level . In the control, the total incidence of heterotrophs decreased 1355 times, that of Gram-negative bacteria fell down 527 times, and that of E . coli cells dropped 3419 times due to the interaction between the host bacteria and Bdellovibrio . The variations in the number of interacting cells were characteristic of a two-component parasite-host system.

Genetika, 1987 Sep, 23(9), 1581 - 7
{Localization of structural genes of Vibrio cholerae cholerae toxin using the recombinant plasmid RP4omega elt}; Evdokimova NM et al.; The recombinant plasmid RP4 omega elt carrying Escherichia coli heat-labile enterotoxin elt genes with 70-80% homology with genes vct of Vibrio cholerae has been constructed . We used this plasmid to determine localization of the cholerae toxin genes vct on the map of Vibrio cholerae cholerae . Two types of the donors were revealed in matings of 10 strains of V . cholerae cholerae 569B/RP4 omega elt with the polyauxotrophic recipients RV31 and RV175: some strains had enhanced frequency of mobilization of ilv-1 and lys-6 markers, the others--of trp-1 . Our data suggest that structural vct genes are located within two regions of V . cholerae cholerae 569B chromosome: trp-1 and ilv-1--lys-6.

Acta Trop, 1987 Sep, 44(3), 273 - 82
Mechanism of cell invasion by Trypanosoma cruzi: importance of sialidase activity; de Titto EH et al.; The sialidase activity of trypomastigotes of Trypanosoma cruzi and its relationship to the ability of different stocks of the organism to infect cultured cells was examined . Sialidase activity in lysates of trypomastigotes was confirmed and shown to be present in organisms of four different stocks of T . cruzi . In addition, sialidase activity was detected in sera of mice acutely infected with organisms of each of the stocks of T . cruzi examined . Erythrocytes from these mice were agglutinated by peanut lectin, suggesting sialidase activity in vivo . Treatment of normal mouse peritoneal macrophages with sera from acutely infected mice resulted in an increased capacity of the cells to internalize blood trypomastigotes . IgM or IgG antibodies specific to T . cruzi were not detected in the sera displaying sialidase activity . Treatment of parasites and/or normal mouse macrophages with Vibrio cholerae neuraminidase, however, had little effect in the rate of internalization of parasites . Treatment of L 929 mouse fibroblasts with neuraminidase reduced significantly the rate of infection of the cells with blood trypomastigotes . Anti-sialidase activity developed and was detected in sera of infected mice and humans, suggesting that the neuraminidase activity of the parasite may play a significant role in the invasion of host cells only during the initial phase of the infection.

Antimicrob Agents Chemother, 1987 Sep, 31(9), 1446 - 9
New tetracycline resistance determinant on R plasmids from Vibrio anguillarum; Aoki T et al.; Two classes of tetracycline resistance determinants on R plasmids were detected in Vibrio anguillarum strains isolated from ayu (sweat fish; Plecoglossus altivelis) farms in Japan . Tetracycline resistance genes categorized as class B were prevalent from 1973 to 1977; however, a new tetracycline resistance gene, which was not classified into tetracycline resistance determinant class A, B, C, or D, has been prevalent since 1981.

Biochim Biophys Acta, 1987 Aug 12, 893(1), 43 - 8
Na+/adenosine co-transport in Vibrio parahaemolyticus; Sakai Y et al.; Adenosine transport in Vibrio parahaemolyticus was studied . Na+ greatly stimulated adenosine uptake . Addition of adenosine to a cell suspension under anaerobic conditions elicited Na+ uptake, and the Na+ uptake was inhibited by monensin, an Na+ ionophore . Imposition of an electrochemical potential of Na+ or a membrane potential in energy-depleted cells elicited adenosine uptake . Therefore, adenosine transport in this organism was concluded to proceed by an Na+/adenosine co-transport mechanism . The Na+/adenosine co-transport system was induced when cells were grown in the presence of adenosine, and repressed by glucose . Although Na+ uptake elicited by adenosine was reduced by glucose, it was enhanced by methyl alpha-glucoside, which reduced the intracellular ATP level . Thus, the effects of glucose and the glucoside on the Na+/adenosine co-transport system did not seem to be due to inducer exclusion, but to be related to the intracellular ATP level.

Biochemistry, 1987 Aug 11, 26(16), 4917 - 21
Polypeptide folding and dimerization in bacterial luciferase occur by a concerted mechanism in vivo; Waddle JJ et al.; Bacterial luciferase is a heterodimeric enzyme comprising two nonidentical but homologous subunits, alpha and beta, encoded by adjacent genes, luxA and luxB . The two genes from Vibrio harveyi were separated and expressed from separate plasmids in Escherichia coli . If both plasmids were present within the same E . coli cell, the level of accumulation of active dimeric luciferase was not dramatically less than within cells containing the intact luxAB sequences . Cells carrying the individual plasmids accumulated large amounts of individual subunits, as evidenced by two-dimensional polyacrylamide gel electrophoresis . Mixing of a lysate of cells carrying the luxA gene with a lysate of cells carrying the luxB gene resulted in formation of very low levels of active heterodimeric luciferase . However, denaturation of the mixed lysates with urea followed by renaturation resulted in formation of large amounts of active luciferase . These observations demonstrate that the two subunits, alpha and beta, if allowed to fold independently in vivo, fold into structures that do not interact to form active heterodimeric luciferase . The encounter complex formed between the two subunits must be an intermediate structure on the pathway to formation of active heterodimeric luciferase.

Infect Immun, 1987 Aug, 55(8), 1936 - 9
Activation of the plasma kallikrein-kinin system by Vibrio vulnificus protease; Miyoshi N et al.; Vibrio vulnificus protease enhanced hypodermic vascular permeability when injected into the dorsal skin of a guinea pig . Enhancement of permeability was observed within 2 min, and the permeability-enhancing reaction terminated at about 10 min postinjection . The permeability-enhancing reaction was greatly augmented by simultaneous injection of a kininase II inhibitor, whereas the reaction was inhibited by soybean trypsin inhibitor, a well-known inhibitor of plasma kallikrein . Furthermore, in vitro activation of plasma prekallikrein to kallikrein by V . vulnificus protease was observed . These results indicate that V . vulnificus protease enhances vascular permeability through activation of the plasma kallikrein-kinin system which generates bradykinin, factor in edema formation.

J Med Microbiol, 1987 Aug, 24(1), 29 - 40
Immunological responses of rabbits to various somatic and secreted antigens of Vibrio cholerae after intra-duodenal inoculation; Kabir S; The immunological responses of rabbits after intra-duodenal immunisation with live Vibrio cholerae organisms were studied in various body fluids . Serum, bile and intestinal samples were collected from rabbits at different times (1-8 weeks) after immunisation . Three different extracts from the small intestine were prepared . At first the small intestine was washed with saline (method A) . Later, ultrasonic lysates were prepared from epithelial cells separated with citrate (method B) and of mucosal tissue above the muscularis layer (method C) . All had agglutinating activities against V . cholerae strains belonging to both biotypes (classical and el tor) and both serotypes (Ogawa and Inaba) . Levels in intestinal extracts, sera and bile of antibodies to somatic (lipopolysaccharide and cell-surface proteins) and secreted (cholera toxin and neuraminidase) antigens of V . cholerae were determined by an enzyme-linked immunosorbent assay . All contained antibodies to these antigens; although both IgA and IgG were present, IgA predominated . Serum and bile samples contained mainly IgG and IgA respectively . Immunoblotting studies demonstrated that the antisera contained antibodies to most cell-surface proteins and to cholera toxin . Cell-surface proteins appeared to be the major cross-reacting somatic antigens of V . cholerae.

Mol Gen Genet, 1987 Aug, 209(1), 175 - 8
Inducible reactivation of UV-irradiated cholera phage e5 in Vibrio cholerae MAK757; Basu R et al.; The survival of UV-irradiated cholera phage e5 was found to increase when the host cells, Vibrio cholerae MAK757, were exposed to a low dose of UV irradiation before phage infection (Weigle reactivation), indicating the existence of a UV-inducible DNA repair pathway (SOS repair) in V . cholerae MAK757 . The induction signal generated by UV irradiation was transient in nature and lasted about 20-30 min at 37 degrees C . Maximal Weigle reactivation of the phage was obtained when the host cells were irradiated with a UV dose of 16 J/m2 . V . cholerae MAK757 was also found to possess efficient photoreactivation and host cell reactivation of UV-damaged DNA in phage e5.

J Gen Microbiol, 1987 Aug, 133 ( Pt 8), 2279 - 84
Monoclonal antibodies against the haemolysin of Vibrio vulnificus; Okada K et al.; The extracellular haemolysin produced by Vibrio vulnificus strain FCC was partially purified from the culture supernate by sequential ammonium sulphate precipitation, gel filtration with Sepharose 4B, and DEAE-Sephacel ion-exchange column chromatography . Using this semi-purified haemolysin as the antigen, several monoclonal antibodies (MAbs) were established; they were all of the IgG2b class with lambda light chains . One representative MAb, 6F8D, completely neutralized the haemolytic activity and mouse lethal activity of extracellular toxin(s) . In immunoblotting analysis of the peptides of the semi-purified haemolysin separated by SDS-PAGE, this MAb reacted, in particular, with a 36 kDa peptide . These findings suggest that the haemolysin is probably identical to the lethal toxin in the culture supernate of V . vulnificus strain FCC, which contained the 36 kDa peptide.

Immunology, 1987 Aug, 61(4), 543 - 7
Specific antibodies to cholera toxin in rabbit milk are protective against Vibrio cholerae-induced intestinal secretion; Yoshiyama Y et al.; Breast feeding helps to protect the nursing infant against infectious diarrhoeas, but the relative importance of antibodies compared with other components present in milk is unsettled . In order to aid in resolving this issue we evaluated the ability of milk, collected from rabbits not immunized or immunized enterally during pregnancy with toxinogenic, live Vibrio cholerae, to inhibit water secretion induced by V . cholerae in rat ileal loops . Non-immune milk was not inhibitory, whereas immune milk was . The inhibitory component of the immune milk was immunoglobulin by virtue of its molecular weight and absorption by an anti-rat immunoglobulin immunosorbent . In addition, the inhibitory antibodies were principally antibodies to cholera toxin because they could be removed from the milk by a cholera toxin immunosorbent but were only partially removed by incubation with whole V . cholerae . Thus, in rabbit milk, we could implicate specific antibodies in protection against intestinal water secretion induced by V . cholerae.

Gut, 1987 Aug, 28(8), 1029 - 32
Single dose tetracycline in cholera; Islam MR; A randomised clinical trial was carried out to explore the efficacy of single dose tetracycline therapy in cholera . One hundred and eighteen adult patients were assigned to receive either tetracycline in a single 1 g, or a single 2 g dose, or tetracycline 500 mg every six hours four times, or no antibiotics as controls . The means of total liquid stool volumes after treatment were lower in the single 1 g dose group (168.0 +/- 20.9 ml/kg), in single 2 g dose group (229.5 +/- 45.6 ml/kg), and multiple dose group (214 +/- 28.5 ml/kg), than in the control group (499.1 +/- 56.5 ml/kg) (p less than 0.05) . Similarly, the means of durations of diarrhoea and intravenous fluid requirements were significantly lower in the single dose and multiple dose tetracycline groups, than in the controls (p less than 0.05) . The mean durations of excretion of Vibrio cholerae were significantly shortened from 3.9 +/- 0.2 days in the control group to 1.9 +/- 0.2 days in single 1 g dose, to 2.2 +/- 0.4 days in single 2 g dose and 1.3 +/- 0.1 days in multiple dose groups, respectively (p less than 0.05) . Three patients in the single 1 g dose group and two patients in single 2 g dose group had clinical relapses with excretion of V cholerae during the relapses, but this was not significantly more frequent than that in the multiple dose group (p greater than 0.05) . These findings suggest that although multiple dose tetracycline therapy remains the best choice, a single dose of either 1 g or 2 g tetracycline appears to be a reasonable alternative for the treatment of cholera as an adjunct to rehydration therapy.

J Bacteriol, 1987 Aug, 169(8), 3785 - 91
Translocation of Vibrio harveyi N,N'-diacetylchitobiase to the outer membrane of Escherichia coli; Jannatipour M et al.; The gene encoding N,N'-diacetylchitobiase (chitobiase) of the chitinolytic marine bacterium Vibrio harveyi has been isolated . While expression of the chitobiase gene (chb) was inducible by N,N'-diacetylchitobiose in V . harveyi, it was expressed constitutively when cloned in Escherichia coli, suggesting that controlling elements are not closely linked to chb . Chitobiase was found in the membrane fraction of E . coli cells containing plasmids with the cloned V . harveyi chb gene . When membranes of such cells were separated on Osborn gradients, chitobiase activity was found mainly in the outer membrane band . Translocation of the enzyme to the outer membrane was accompanied by cleavage of a signal peptide . A fusion protein, in which 22 amino acids from the amino terminus of prechitobiase were replaced with 21 amino acids from the pUC19 lacZ amino terminus, was not processed, and 99% of the activity was located in the cytoplasmic fraction . A homology to six amino acids surrounding the lipoprotein processing and modification site was found near the amino terminus of prechitobiase.

J Bacteriol, 1987 Aug, 169(8), 3441 - 9
Phosphate regulation of gene expression in Vibrio parahaemolyticus; McCarter LL et al.; The synthesis of a major outer membrane protein, OmpP, in Vibrio parahaemolyticus was induced by growth in media deficient in phosphate . The gene, ompP, encoding this protein was cloned . Synthesis of OmpP in Escherichia coli was regulated by the availability of phosphate, and this control required the function of pho regulatory genes of E . coli . Analysis of gene fusion strains constructed by mutagenesis with transposon mini-Mulux revealed that ompP was transcriptionally regulated in V . parahaemolyticus . Impaired growth of a strain with an ompP defect was observed in media which contained large linear polyphosphates as the phosphate source . This and other evidence suggested that OmpP functions as a porin channel for the entry of phosphate into the cell . A number of other proteins or activities were induced by phosphate limitation including hemolysin, phospholipase C, and phosphatase activities . A regulatory locus controlling expression of phosphate-regulated genes was identified and cloned . This regulatory locus cloned from V . parahaemolyticus was shown to complement E . coli strains with defects in pho regulatory genes.

Jpn J Med Sci Biol, 1987 Aug, 40(4), 153 - 7
Vibrio fluvialis: a new serogroup (19) possessing the Inaba factor antigen of Vibrio cholerae O1; Shimada T et al.; A serogroup of Vibrio fluvialis possessing the C (Inaba) antigen but not the B (Ogawa) nor A antigen of V . cholerae O1 is described . The O-antigen of this serogroup was identical with that of bioserogroup 1875-variant of a marine Vibrio species . As the O-antigen of this serogroup was not agglutinated by any of O-antisera for the 18 serogroups of V . fluvialis already recognized, it was designated O-serogroup 19 of this species.

Biochem Biophys Res Commun, 1987 Jul 15, 146(1), 101 - 6
Purification of the yellow fluorescent protein from Vibrio fischeri and identity of the flavin chromophore; Macheroux P et al.; A low molecular weight protein (approximately 25,000 D) exhibiting a yellow fluorescence emission peaking at approximately 540 nm was isolated from Vibrio fischeri (strain Y-1) and purified to apparent homogeneity . FMN is the chromophore, but it exhibits marked red shifts in both the absorption (lambda max = 380, 460 nm) and the fluorescence emission . When added to purified luciferase from the same strain, which itself catalyzes an emission of blue-green light (lambda max approximately 495 nm), this protein induces a bright yellow luminescence (lambda max approximately 540 nm); this corresponds to the emission of the Y-1 strain in vivo . This yellow bioluminescence emission is thus ascribed to the interaction of these two proteins, and to the excitation of the singlet FMN bound to this fluorescent protein.

J Gen Microbiol, 1987 Jul, 133 ( Pt 7), 1861 - 70
The role of osmotic effects in haloadaptation of Vibrio costicola; Adams R et al.; Growth rates of Vibrio costicola showed a broad optimum between 0.8 and 1.5 M-NaCl, and there was no growth above 3.3 M-NaCl in a peptone-based medium . The minimum requirement of 0.5 M-NaCl for growth in NaCl alone was reduced to 0.3 M-NaCl when the total solute concentration was raised to 0.5 to 1.0 M equivalent with sucrose or glycerol . Compared with equivalent NaCl concentrations, higher concentrations of sucrose were more inhibitory to growth, whereas glycerol had less effect . Increasing the medium NaCl concentration suddenly by 2- or 3-fold with either a constant starting, or final, salt concentration showed that, after the shift-up, the lag in growth, the rate of growth, and the inhibition of phospholipid synthesis depended both on the final NaCl concentration and the magnitude of the shift in salinity . The time-courses of phospholipid synthesis following a 2- or 3-fold shift-up in NaCl or sucrose media were very similar and exhibited a relative increase in phosphatidylglycerol synthesis over that of phosphatidylethanolamine . This 'switch-over' was not seen following shift-up in glycerol media when there was also a stimulation, rather than inhibition, of phospholipid synthesis . It is concluded that during phenotypic haloadaptation of V . costicola, osmotic effects play a significant part in the sensing of and response to raised external salinity.

Appl Environ Microbiol, 1987 Jul, 53(7), 1556 - 9
Use of sodium dodecyl sulfate-polymyxin B-sucrose medium for isolation of Vibrio vulnificus from shellfish; Bryant RG et al.; The differential and selective sodium dodecyl sulfate-polymyxin B-sucrose medium (SPS) of Kitaura et al . (T . Kitaura, S . Doke, I . Azuma, M . Imaida, K . Miyano, K . Harada, and E . Yabuuchii, FEMS Microbiol . Lett . 17:205-209, 1983), which highlights alkylsulfatase activity, was evaluated for its potential use in the direct isolation and enumeration of Vibrio vulnificus from shellfish . V . vulnificus was detected by this method in six of nine shellfish samples collected from diverse geographic locales during the summer of 1986 . Direct enumeration of V . vulnificus at 7.0 X 10(2) to 2.2 X 10(4) CFU/g of shellfish was achieved on SPS agar . All sample results were confirmed in parallel examinations by using conventional glucose-salt-Teepol (Shell Oil Co.) broth and alkaline peptone water enrichment with plating onto thiosulfate-citrate-bile salts-sucrose agar . Additionally, alkylsulfatase activity was evaluated in vitro for 97 strains representing 14 Vibrio spp . V . vulnificus and Vibrio cholerae-01 were the only species consistently found to possess this activity . The range of plating efficiencies for random V . vulnificus strains analyzed on SPS was 11 to 74% (mean, 39%) . The use of SPS shows great promise for the study of shellfish and other environmental sources for V . vulnificus.

Food Addit Contam, 1987 Jul-Sep, 4(3), 291 - 6
Oxytetracycline residues in rainbow trout analysed by a rapid HPLC method; Nordlander I et al.; A rapid and sensitive HPLC method was developed for the determination of oxytetracycline in fish tissues (muscle and liver) based on a clean-up and concentration procedure on Sep-Pak C18 . At a coastal fishfarm rainbow trout (Salmo gairdneri) suffering from Vibrio anguillarum were treated with 75 mg oxytetracycline per kg fish and day for ten days . Oxytetracycline residues above the limit of determination (0.005 micrograms/g) were found in fish 82 days after treatment . The recoveries from spiked tissues were about 60% and 70% for muscle and liver, respectively.

Res Vet Sci, 1987 Jul, 43(1), 78 - 84
Detection of fish antibody against protein antigen of Aeromonas salmonicida by enzyme-linked immunosorbent assay using biotin-avidin system; Kodama H et al.; Antibody against Aeromonas salmonicida was detected in sera from immunised or experimentally infected rainbow trout by enzyme-linked immunosorbent assay (ELISA) using the biotin-avidin system . The ELISA titre correlated well with the agglutinin titres of the sera, but the ELISA was found to be more sensitive than the agglutination test . When the rainbow trout serum was separated by column chromatography, antibody activity (determined by ELISA and agglutination test) was detected in the IgM fractions . Minimum cross reaction was observed in the ELISA system between antigen prepared from A salmonicida and antibodies against Vibrio species and other species of Aeromonas . The specificity of the ELISA was also confirmed by inhibition test . Immunisation of rainbow trout with a virulent strain of A salmonicida provided good protection, though no correlation was observed between the protection and the ELISA titres of sera.

Trop Geogr Med, 1987 Jul, 39(3), 271 - 5
Vibriocidal titre in cholera cases and contacts: its value in assessing endemicity of or susceptibility to cholera; Khan MU et al.; Vibriocidal antibody titre in excess of 1:40 occurred within two weeks of cholera infection, both in severe hospitalized cases, contact cases and in asymptomatic infected contacts . These levels, considered to be indicative of protection, persisted for six months or longer in more than half of the subjects irrespective of presence and severity of symptoms . Approximately 40% of infected family contacts had similar titres implying recent infection and subsequent protection . The use of antibiotics to treat acute cases, and whether infection was due to antibiotic resistant or sensitive Vibrio cholerae had no effect on the response of vibriocidal titre . Endemicity of cholera was higher than previously observed in Dhaka . Screening populations to obtain positive titre rates permits retrospective assessment of cholera infection and provides an indicator of future susceptibility.

J Gen Microbiol, 1987 Jul, 133 ( Pt 7), 1783 - 91
Purification and characterization of an elastolytic protease of Vibrio vulnificus; Kothary MH et al.; Large amounts of a highly purified, extracellular elastolytic protease of Vibrio vulnificus were obtained by sequential ammonium sulphate precipitation and hydrophobic interaction chromatography with phenyl-Sepharose CL-4B . The protease had an Mr of about 50,500 (estimated by SDS-PAGE), a pI of 5.7, and a temperature optimum range of 55 to 60 degrees C . The pH optimum and the results of inactivation studies suggested that the enzyme was a neutral metalloprotease . The protease had about 429 amino acid residues, and the first 20 amino-terminal amino acid residues were Ala-Gln-Ala-Asn-Gly-Thr-Gly-Pro-Gly-Gly-Asn-Ser-Lys-Thr-Gly-Arg-Tyr-Glu- Phe-Gly . The purified protease was toxic for mice (about 1.5 mg kg-1 and 4.5 mg kg-1, intraperitoneal and intravenous LD50 values, respectively), and subcutaneous injection of the enzyme elicited rapid and extensive dermonecrosis.

Appl Environ Microbiol, 1987 Jul, 53(7), 1714 - 5
Marine bacteria which produce tetrodotoxin; Simidu U et al.; A number of type strains of marine bacteria, including members of the family Vibrionaceae, were cultured and examined for tetrodotoxin productivity by high-performance liquid chromatography and gas chromatography-mass spectrometry . Most of the Vibrionaceae strains produced tetrodotoxin, anhydrotetrodotoxin, or both.

J Neurochem, 1987 Jul, 49(1), 201 - 7
Effect of exogenous gangliosides on amino acid uptake and Na+, K+-ATPase activity in superior cervical and nodose ganglia of rats; Nagata Y et al.; The effects of some gangliosides on active uptake of nonmetabolizable alpha-aminoisobutyric acid (AIB) and Na+, K+-ATPase and Ca2+, Mg2+-ATPase activities in superior cervical ganglia (SCG) and nodose ganglia (NG) excised from adult rats were examined during aerobic incubation at 37 degrees C for 2 h . In NG, amino acid uptake was greatly accelerated with the addition of galactosyl-N-acetylgalactosaminyl-{N-acetylneuraminyl}-galactosylgluc osyl ceramide (GM1) (85%) and also with N-acetylgalactosaminyl-{N-acetylneuraminyl}-galactosylglucosyl ceramide (GM2) or {N-acetylneuraminyl}-galactosyl-N-acetylgalactosaminyl-{N-acetyl- neuraminyl}-galactosylglucosyl ceramide (GD1a) (43% each) compared with a nonaddition control at a 5 nM concentration . Under identical conditions, Na+, K+-ATPase activity was strongly stimulated with GM1 (180%) and GD1a (93%), whereas Ca2+, Mg2+-ATPase activity showed no change . In SCG, on the other hand, AIB uptake was apparently inhibited (-27%) by addition of GM1, with a slight decrease in Na+, K+-ATPase but no change in Ca2+, Mg2+-ATPase activity in the tissue . Both asialo-GM1, in which N-acetylneuraminic acid is deficient, and Forssman glycolipid, which is not present in nervous tissue, failed to produce any significant increase in both SCG and NG not only in amino acid uptake, but also in Na+, K+-ATPase activity . A kinetic study of active AIB uptake showed that GM1 ganglioside produced an increase in Km with no change in Vmax in SCG, whereas it caused a decrease in Km with a slight increase in Vmax in NG . Treatment of NG and SCG with neuraminidase from Vibrio cholerae, an enzyme that split off sialic acid from polysialoganglioside, leaving GM1 intact, caused little inhibition of the amino acid uptake.(ABSTRACT TRUNCATED AT 250 WORDS)

Zh Mikrobiol Epidemiol Immunobiol, 1987 Jul, (7), 28 - 31
{Factors in the spread of diseases caused by nonagglutinating vibrios studied by using the data from strain serotyping}; Zaidenov AM et al.; The method of the serotyping of strains was used for the epidemiological evaluation of the role of different factors in the transfer of infective agents in 147 cases of diseases and carrier state, caused by NAG vibrios, in Karakalpakia . Out of 150 NAG vibrio strains, 136 strains were serotyped and classified with 26 serovars . The strains were found to belong mostly to serovars 47, 37, 5 and 6 (42.0%) . Most of the infected persons (58.5%) used water from open water bodies for household purposes . The role of water factor in the spread of infection was confirmed by a wide spectrum of serologic variants, a low index of the focal outbreaks (1.02), and sporadic pattern of infection . No group morbidity or toxinfection-type outbreaks were recorded.

Plasmid, 1987 Jul, 18(1), 1 - 7
Characterization and restriction analysis of the P sex factor and the cryptic plasmid of Vibrio cholerae strain V58; Bartowsky EJ et al.; The P plasmid of Vibrio cholerae is a derepressed sex factor restricted to V . cholerae and has been shown to express surface exclusion . We have isolated the plasmids of strain V58 and have found that in addition to P, two further cryptic plasmids are also present . P has a size of 68 kb as determined by both electron microscopy and restriction endonuclease analysis . These other plasmids are 34 and 4.7 kb in size . Restriction maps of P and the larger cryptic plasmid have been determined . It has been demonstrated that P differs from the standard Inc group test plasmids and also expresses a surface exclusion system . The ability of the type Inc plasmids to be transferred to V . cholerae by either liquid or filter matings and the stability of these plasmids in V . cholerae have also been examined.

Biochim Biophys Acta, 1987 Jun 22, 924(3), 393 - 402
Vibrio cholerae metalloproteinase degrades intestinal mucin and facilitates enterotoxin-induced secretion from rat intestine; Crowther RS et al.; Mucin secretion in situ from rat intestinal loops was promoted more effectively by dialysed crude cholera filtrate than by an equivalent amount of purified enterotoxin . The filtrate could be rendered inactive by incubation with mixed gangliosides or passage through a GM1-affinity column, which indicated that the secretory action of the filtrate depended upon its enterotoxin component . In an effort to explain the greater potency of the filtrate, we established the presence of a metalloproteinase in the filtrate and demonstrated that this enzyme was capable of degrading purified rat intestinal mucin . Sufficient degradation occurred to cause a substantial decrease in viscosity (57% in 120 min) . Biochemical analysis of the mucin before and after exposure to filtrate revealed a rise in the combined percentage of serine, threonine and proline (53-58%), suggesting that poorly glycosylated areas (which are less abundant in these amino acids) were being partly removed from the mucin . The carbohydrate composition was essentially unaltered . Inhibition of the filtrate metalloproteinase by Zincov and alpha 2-macroglobulin significantly (P less than 0.005) reduced the ability of cholera filtrate to degrade mucin or to stimulate mucin secretion from rat intestinal slices in vitro . Purified cholera enterotoxin added to enterotoxin-depleted filtrate was a more potent secretagogue (secretory stimulant) in intestinal loops than an equivalent amount of enterotoxin alone . We therefore propose that mucin secretion induced by cholera filtrate is caused by cholera enterotoxin, but that degradation of the protective epithelial mucus layer by a constituent metalloproteinase may assist the toxin by allowing increased access to mucosal GM1 receptor sites.

Appl Environ Microbiol, 1987 Jun, 53(6), 1349 - 51
Virulent strains of Vibrio vulnificus isolated from estuaries of the United States West Coast; Kaysner CA et al.; Vibrio vulnificus was isolated from United States West Coast estuaries at a low frequency (5.9%) from 529 samples of water, shellfish, and sediment . Four strains tested with iron-treated mice had 50% lethal dose values ranging from 7.6 to 360 CFU, compared with a 50% lethal dose of 4.9 CFU for a clinical isolate that caused the death of a septicemic patient . The presence of this pathogen may be a hazard to users of marine beaches and consumers of raw shellfish on the West Coast, especially to persons most susceptible to V . vulnificus septicemia . Species-specific antiflagellar serum and a gene probe for cytotoxin-hemolysin production were useful for screening these environmental isolates.

Appl Environ Microbiol, 1987 Jun, 53(6), 1344 - 8
Incidence of Vibrio cholerae from estuaries of the United States West Coast; Kaysner CA et al.; The incidence of Vibrio cholerae in shellfish, sediment, and waters of California, Oregon, and Washington was determined during the summer of 1984 . Samples from 24 distinct estuaries were analyzed qualitatively . V . cholerae non-O1 was found in 23 estuaries and in 44.6% of the 529 samples examined . V . cholerae O1 Inaba was isolated from water samples in Morro Bay, Calif . Vibrio mimicus was found in 2.3% of the samples . Cholera enterotoxin was not found in cell-free filtrates of the 100 isolates tested in the Y-1 mouse adrenal cell assay, but heat-labile cytotoxic activity was observed with 3% of the isolates.

J Med Microbiol, 1987 Jun, 23(4), 331 - 4
Phage-induced change of toxigenesis in Vibrio cholerae; Siddiqui KA et al.; A temperate phage coding for constitutive hypertoxigenicity has been constructed in Vibrio cholerae strain 569B and used to lysogenise the low-toxin-producing strain MAK 757; 18% of lysogens showed 10-100-fold increase in toxin production . This property was also transmitted at low frequency to second generation lysogens . Thus temperate phage can increase toxin production in a low-toxin-producing strain.

Ann Emerg Med, 1987 Jun, 16(6), 643 - 9
Bacteriology of the marine environment: implications for clinical therapy; Auerbach PS et al.; Ocean water and tissue samples were obtained from a variety of sources with phylogenetic and geographic diversity . Purified bacterial colonies were isolated and identification procedures were performed . A total of 67 isolates were recovered . Thirty-eight isolates belonged to the genus Vibrio and included six species . Twenty-four non-fermentative bacteria and four Gram-positive isolates were recovered . Antibiotic susceptibility testing showed that while the non-fermentative marine bacteria generally were susceptible to the antibiotics tested, marine Vibrio species were relatively resistant to a wide variety of antimicrobials . Antibiotics effective against all species included imipenem, trimethoprim/sulfamethoxazole, and chloramphenicol . Further recommendations for treatment are based on sensitivity in culture . Some isolates failed to grow in the medium used for susceptibility testing . Because commercial test kits may not yield accurate identifications of bacteria, the acquisition of antimicrobial susceptibility data gains added importance.

Infect Immun, 1987 Jun, 55(6), 1529 - 32
Transcription of cholera toxin operon in wild-type and mutant strains of Vibrio cholerae; Mishra L et al.; mRNA for cholera toxin (CT) was measured in wild-type, hypertoxinogenic, and hypotoxinogenic strains of the classical biotype . The amount of CT-specific mRNA was directly related to the amount of CT that the strains could produce . Estimates of the size of the mRNA for CT in individual experiments varied between approximately 900 and 1,100 nucleotides.

Biull Eksp Biol Med, 1987 Jun, 103(6), 717 - 9
{Isolation of the R'his plasmids of Vibrio cholerae}; Rusina OIu et al.; V . cholerae strain VT5104 capable of donor activity in conjugation has been constructed by the genetic technique based on plasmid RP4::Mucts62 integration into V . cholerae chromosome due to plasmid homology with Mucts62 inserted into the chromosome . The gene for histidine synthesis has been mobilized and transferred into the recipient cells from VT5104 donor . The conjugants obtained are able to efficiently transfer his+ gene included into the plasmid structure in conjugation with eltor recipient . Thus, the constructed strain VT5104 generates R' plasmids carrying V . cholerae chromosomal genes.

Southeast Asian J Trop Med Public Health, 1987 Jun, 18(2), 142 - 8
Oral vaccine against cholera prepared from Vibrio cholerae antigen(s); Chaicumpa W et al.; Albino rats aged 7-8 weeks old purchased from the National Laboratory Animal Centre, Salaya, Nakhon Pathom, were found to be a good animal model for the study on immunogenicity of V . cholerae antigens . Seventy-two rats were fasted for 15 hours before feeding each one with 1 ml of 5% NaHCO3 to reduce gastric acidity prior to immunization . They were divided into 9 groups of 8 rats and immunized orally with 2 ml, each, of the V . cholerae antigens dissolved or suspended in Cassamino acid as follows: group 1 (control): Cassamino acid (Ca) alone; group 2 (control): 2.5% formalinized sheep red blood cells (F-SRBC); group 3: 1,000 micrograms of lipopolysaccharide (LPS); group 4: 100 micrograms of procholeragenoid (P); group 5: 80 haemagglutinating units of cell-bound haemagglutinin (CHA) adsorbed onto the surface of F-SRBC (CH-SRBC); group 6: 500 micrograms of LPS + 50 micrograms of P; group 7: CH-SRBC + 50 micrograms of P; group 8: combined vaccine formula 1 consisted of 500 micrograms of LPS, CH-SRBC and 50 micrograms of P and group 9: combined vaccine formula 2 consisted of 1,000 micrograms of LPS, CH-SRBC and 100 micrograms of P . The immunization was repeated once more 14 days later . Five days, thereafter, the rats were killed and their jejuni were removed for cryostat sectioning . Antibody producing cells against LPS (anti-LPS cells), P (anti-CT cells) and CHA (anti-CHA cells) in the intestinal lamina propria were enumerated by double antibody sandwich method of immunofluorescence using pure LPS, cholera toxin (CT) and pure CHA as the antigens in the assay, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Zh Mikrobiol Epidemiol Immunobiol, 1987 Jun, (6), 3 - 6
{Use of a new method for restoring the type properties of atypical Vibrio cholerae}; Pasternak NA et al.; Subcultures with a number of signs characteristic of epidemically significant strains have been isolated from cholera vibrios, nonpathogenic and atypical in a number of properties, by a new in vitro method developed by the authors . This method makes it possible to increase the virulence of poorly agglutinating cultures of V . cholerae O1 and their agglutinability with cholera antisera.

Mol Cell Biochem, 1987 Jun, 75(2), 103 - 11
Structural and immunochemical characterization of the O-haptens of Brucella abortus lipopolysaccharides from strains 19 and 2308; Wu AM et al.; The O-haptens of the major fraction (f5A) of B . abortus (Strains 2308 and 19) membrane bound smooth lipopolysaccharide (sLPS) were prepared by hydrolysis of f5A native sLPS in 1% acetic acid at 100 degrees C for 2 h . After hydrolysis, O-haptens were separated from Lipid A-protein complex by centrifugation, and from small fragments by ultrafiltration of molecular weight cut-off (MWCO) 1.0 X 10(3) . These carbohydrate haptens were identified by precipitin-inhibition assay and further fractionated by both membrane filtration and dialysis . The size distributions of carbohydrate haptens of endotoxins (f5A) ranged from oligosaccharides up to polysacchandes of 1.0 X 10(4) MWCO . Three major fractions of MWCO 8.0-10.0 X 10(3), 3.5-5.0 X 10(3) and less than 1.0 X 10(3) from both strains 2308 and 19 contained more than 85% of the total immunoactive materials . These fractions of haptens were subjected to composition, proton and 13C NMR analysis and were found to be a homopolymer of alpha 1----2 linked, 4,5-dideoxy-4-formamido-D-mannose (N-formylperosamine), which is identical to O-haptens of B . abortus strain 119.3 and Yersinia enterocolitica serotype 0:9 and similar to Vibrio cholerae 569B (INABA) . Fractions of these haptens exhibited similar inhibitory reactivities in a precipitin-inhibition assay as expressed as mumoles of monosaccharide of anhydro-N-formyl perosamine . They were about 480 times as active as Me alpha----DMan or DMan.

J Bacteriol, 1987 Jun, 169(6), 2685 - 90
Expression and regulation of a Vibrio alginolyticus sucrose utilization system cloned in Escherichia coli; Scholle RR et al.; A halotolerant collagenolytic Vibrio alginolyticus strain isolated from salted hides had intracellular sucrase activity and did not secret sucrase into the medium . The strain actively transported sucrose by a sucrose-inducible, Na+-independent process . A 10.4-kilobase DNA fragment of V . alginolyticus DNA was cloned into Escherichia coli . The recombinant E . coli(pVS100) could utilize sucrose as a sole carbon source . In contrast to V . alginolyticus, the recombinant E . coli produced both intra- and extracellular sucrase activities . Up to 20% of the total sucrase activity was in the supernatant . Sucrase synthesis in E . coli(pVS100) was inducible and was subject to glucose repression, which was relieved by cyclic AMP . Sucrose was actively transported by a sucrose-inducible, Na+-independent system in E . coli(pVS100) . Sucrose uptake was inhibited by the addition of a proton conductor . The maximum velocity and apparent Km values of sucrose uptake for the V . alginolyticus strain and E . coli(pVS100) were 130 nmol/mg of protein per min and 50 microM and 6 nmol/mg of protein per min and 275 microM, respectively.

Vaccine, 1987 Jun, 5(2), 83 - 7
Involvement of cell envelope components in the pathogenesis of Vibrio cholerae: targets for cholera vaccine development; Manning PA; Parenteral cholera vaccines have for some years been removed from the WHO recommendations . With a greater understanding of the immunology of gut infections has come the realization that oral vaccines are the most promising approach . There are several possible approaches: killed vaccines, attenuated Vibrio cholerae and true recombinants in which V . cholerae protective antigens are introduced into suitable carrier strains . This latter approach has perhaps greater potential as a carrier strain can be chosen/developed which most effectively stimulates the local immune response in the gut . This necessitates the identification of the important protective antigens . These antigens are thought to be components of the cell envelope which are involved in adhesion and colonization.

J Clin Lab Immunol, 1987 Jun, 23(2), 91 - 4
Contradictory responses in induction of delayed type hypersensitivity in orally immunized mice; Tsuru S et al.; Induction of delayed type hypersensitivity (DTH) in mice fed with sheep red blood cells (SRBC) and live Vibrio cholerae (V . cholerae) both forcedly and ad lib was comparatively investigated . Although suppression of DTH to SRBC or V . cholerae was induced in mice fed forcedly for 1 or 2 weeks, mice fed ad lib could produce the positive footpad reactions to antigens . Furthermore, the suppression of DTH in forcedly fed mice showed an antigenic specificity . These observations indicated that the induction of unresponsiveness to DTH in orally immunized mice was markedly influenced by the oral administration.

J Natl Cancer Inst, 1987 Jun, 78(6), 1169 - 75
Epiglycanin-immunoreactive glycoproteins in mouse fetal tissues and fetal cells in culture; Beppu M et al.; Fetal tissues from time-pregnant female A/J mice of 16- and 19-day pregnancies and from neonates 1 day after birth, as well as from fetal cells in culture, absorbed significant amounts of anti-epiglycanin antibody . Detergent-solubilized glycoproteins, with epiglycanin activity, from fetal tissues and cells were separated by polyacrylamide gel electrophoresis, and the protein bands electroblotted onto a nitrocellulose gel . After the antigens were labeled with rabbit anti-epiglycanin antiserum and {125I}epiglycanin, autoradiography revealed two major bands containing the antigenic determinant at Mr 90,000 and 82,000 . Bands of similar molecular weights, but with no demonstrated immunologic cross-reactivity, were observed by fluorography, if intact cells prior to solubilization were labeled by galactose oxidase followed by sodium borotritiide . Immunoreactive epiglycanin activity could be destroyed by Pronase, endo-N-acetyl-alpha-D-galactosaminidase (Diplococcus pneumoniae), or periodate oxidation . Activity was enhanced with neuraminidase . The spleen, liver, or erythrocytes from adult A/J mice did not possess the antigen, but incubation of adult spleen or liver with neuraminidase (Vibrio cholerae) exposed the epitope.

Anal Biochem, 1987 May 15, 163(1), 123 - 35
Sialyltransferases as specific cell surface probes of terminal and penultimate saccharide structures on living cells; Whiteheart SW et al.; Rat liver beta-galactoside alpha-2,6-sialyltransferase and Vibrio cholerae sialidase were used, in conjunction with CMP-N-acetyl-{3H}neuraminic acid, to probe the glycoconjugate distribution, sialylation state, and level of penultimate Gal beta 1-4GlcNAc residues on the surfaces of murine thymic lymphocytes . We report a detailed characterization of this sialyltransferase-mediated labeling system . Exogenous sialylation of intact cells is dependent on transferase, sugar nucleotide donor, cell number, and incubation time . Additionally, we have demonstrated that the system labeling the cell surface is noncytotoxic and nonmetabolic and is interacting with the entire cell population . Analysis of the exosialylated structures indicates that the sialyltransferase specifically produces an alpha 2-6 linkage on N-linked oligosaccharides . Using this labeling system, we have probed the cell surface saccharide structures of murine thymocytes and demonstrated that most Gal beta 1-4GlcNAc residues are sialylated in the native state . However, one antigen, T200 (Ly-5), is strikingly undersialylated when compared to other cell surface glycoproteins (e.g., Thy 1.2) . Upon analysis of exogenously sialylated oligosaccharides, labeled sialic acid was found almost exclusively on monosialylated structures with the remainder on bisialylated oligosaccharides . This suggests that the purified sialyltransferase is very precise in its recognition of oligosaccharides present on the surface of living thymic lymphocytes . This paper illustrates the combined uses of specific glycosidases and glycosyltransferases and how they can be employed in the detailed study of selected cell surface saccharide structures on living nucleated cells.

FEBS Lett, 1987 May 11, 215(2), 335 - 8
Conjugation-dependent recovery of the Na+ pump in a mutant of Vibrio alginolyticus lacking three subunits of the Na+ pump; Tokuda H et al.; The Na+ pump-deficient mutant, Nap1, of Vibrio alginolyticus was found to lack three subunits of Na+-dependent NADH:quinone oxidoreductase complex . Although a spontaneous Na+ pump positive revertant did not appear from Nap1, transconjugants that recovered both the Na+ pump activity and the subunits were isolated from Nap1 conjugated with the wild type . Moreover, the wild type was found to contain two different sizes of plasmids . These results suggest the possibility that the Na+ pump is encoded by a plasmid.

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1987 May, 20(2), 140 - 7
{Contamination of seafood by Vibrio parahaemolyticus in Taiwan}; Fang SW et al.; From October 1984 to September 1985, a total of 770 seafood samples, collected from the retail markets of 8 coastal cities in Taiwan, were tested for the contamination of Vibrio parahaemolyticus . The results showed that 352 samples (45.7%) were contaminated by V . parahaemolyticus . The detection frequencies of various samples were as follows: 40.0% fish samples, 22.3% fish fillets, 44.4% shrimps and 47.8% crabs of the crustacea group, 68.7% bivalve shellfish and 31.9% non-bivalve shellfish of the mollusca group . Bivalve shellfish samples showed the highest detection frequency and counts . By the analysis of variance, cities and sample items affected the detection frequency and counts of V . parahaemolyticus . Positive statistical correlation was found between the counts of V . parahaemolyticus and temperature variation in some cities, whereas the detection frequency of V . parahaemolyticus was not relative to the temperature variation in each city . 182 isolates from different types of seafood reacted with K antisera . Of those serotypes, K17 showed the highest detection frequency in the serological test . All of our isolates showed negative reaction for the Kanagawa test.

Appl Environ Microbiol, 1987 May, 53(5), 1203 - 5
Evaluation of the multitest medium for rapid presumptive identification of Vibrio cholerae from environmental sources; Nair GB et al.; The multitest V . cholerae medium (VC medium) for rapid presumptive identification of Vibrio cholerae was evaluated . On the basis of reactions in the VC medium, 379 strains recovered during a yearlong ecological study in Calcutta were presumptively identified as V . cholerae . Further phenotypic characterization of these strains revealed that the reactions of 371 (97.9%) isolates were consistent with that of V . cholerae . False-positive reactions were exhibited by eight (2.1%) strains, three of which were identified as Vibrio fluvialis biotype 1 . By slightly varying the basic formulation of the VC medium, we could eliminate some false-positive reactions . On the basis of the present evaluation, we recommend the routine use of the VC medium.

Appl Environ Microbiol, 1987 May, 53(5), 1181 - 2
Elevated temperature method for recovery of Vibrio cholerae from oysters (Crassostrea gigas); DePaola A et al.; Of 222 Vibrio cholerae isolates from diverse clinical and environmental sources, 219 produced visible growth in alkaline peptone broth when incubated overnight at 42 degrees C . In field trials conducted to compare enrichment at incubation temperatures of 42 and 35 degrees C, significantly higher rates of isolation (P less than 0.05) and recovery (P less than 0.01) of V . cholerae from oysters were observed at 42 degrees C.

J Clin Microbiol, 1987 May, 25(5), 900 - 6
Aeromonas veronii, a new ornithine decarboxylase-positive species that may cause diarrhea; Hickman-Brenner FW et al.; In 1983, the vernacular name Enteric Group 77 was coined for a group of strains that had been referred to our laboratory as "possible Vibrio cholerae except for gas production." By DNA-DNA hybridization (hydroxyapatite, 32P), 8 of 10 strains of Enteric Group 77 were very highly related to the labeled strain 1169-83 (74 to 100% at 60 degrees C and 75 to 100% at 75 degrees C; percent divergence, 0.0 to 2.5) . Type strains of six other Aeromonas species were 45 to 66% related (60 degrees C) to strain 1169-83, but type strains of 27 Vibrio species were only 2 to 6% related . The name Aeromonas veronii is proposed for the highly related group of nine strains formerly known as Enteric Group 77 . The type strain is designated as ATCC 35604 (CDC 1169-83) . Strains of A . veronii grew well at 36 degrees C and had positive reactions at this temperature for indole, methyl red, Voges-Proskauer, citrate, lysine and ornithine decarboxylases, DNase, lipase, and motility; the strains had negative reactions for arginine decarboxylase, H2S, urea, and malonate . The following sugars were fermented: D-glucose (acid and gas), cellobiose (seven of nine strains), D-galactose, maltose, D-mannitol, D-mannose, alpha-methyl-D-glucoside (eight of nine strains), salicin, sucrose, and trehalose . The following sugars were not fermented: adonitol, L-arabinose, D-arabitol, dulcitol, erythritol, myo-inositol, lactose, raffinose, L-rhamnose, D-sorbitol, and D-xylose . The positive ornithine decarboxylase reaction differentiates A . veronii from other Aeromonas species . The antibiogram of A . veronii is typical of other Aeromonas strains (resistance to ampicillin and carbenicillin and susceptibility to most other agents) . A . veronii strains were isolated from three clinical sources: respiratory secretions of four victims of drowning or near drowning in fresh water (probably not clinically significant); infected wounds of two patients previously exposed to fresh water (unknown clinical significance); and stools from three patients with diarrhea (probably clinically significant).

J Virol, 1987 May, 61(5), 1407 - 15
Inhibition of reovirus type 3 binding to host cells by sialylated glycoproteins is mediated through the viral attachment protein; Pacitti AF et al.; The interaction of mammalian reoviruses with sialylated glycoproteins was studied and found to be highly serotype specific in that attachment of type 3 Dearing reovirus to murine L cell receptors could be strongly inhibited by bovine submaxillary mucin (BSM), fetuin, and alpha 1 acid glycoprotein, albeit at different efficiencies, whereas attachment of type 1 Lang reovirus was inhibited only by fetuin . We subsequently demonstrated, by using reassortants between type 3 and 1 reoviruses, that inhibition of reovirus attachment to cell receptors was specified by the viral attachment protein gene S1 . Using a solid-phase binding assay, we further demonstrated that the ability of reovirus type 3 or reassortant 1HA3 and the inability of reovirus type 1 or reassortant 3HA1 to bind avidly to BSM was a property of the viral S1 genome segment and required the presence of sialic acid residues on BSM oligosaccharides . Taken together, these results demonstrated that there is a serotype-specific difference in the ability of the reovirus attachment protein, sigma 1, to interact with sialylated oligosaccharides of glycoproteins . Interaction of reovirus type 3 with sialylated oligosaccharides of BSM is dramatically affected by the degree of O-acetylation of their sialic acid residues, as indicated by the findings that chemical removal of O-acetyl groups stimulated reovirus type 3 attachment to BSM, whereas preferential removal of residues lacking or possessing reduced amounts of O-acetyl groups per sialic acid molecule with Vibrio cholerae sialidase abolished binding . We also demonstrated that BSM was 10 times more potent in inhibiting attachment of infectious reovirus to L cells than was V . cholerae-treated BSM . The results are consistent with the hypothesis that sialylated oligosaccharides on host cells or erythrocytes may act as binding sites or components of binding sites for type 3 reovirus through a specific interaction with the virus attachment protein.

J Gen Virol, 1987 May, 68 ( Pt 5), 1411 - 6
Characterization of Vibrio eltor typing phages: properties of the Eltor phage e4; Chattopadhyay S et al.; Biophysical characteristics of Vibrio eltor phage e4, a key phage in the Vibrio cholerae typing scheme were studied . This icosahedral phage was found to contain 12 structural polypeptides with mol . wt . ranging from 25,000 to 120,000 . One of these polypeptides of mol . wt . 50,000 accounted for most of the structural proteins present and was probably the major phage capsid protein . The phage genome comprised a single linear, double-stranded DNA molecule, 69.2 kbp in length (45.6 X 10(6) mol . wt.) as determined by electron microscopy and restriction fragment analyses . The G + C content was 34.6% . Electron microscopy data indicated that unlike the DNAs of other cholera phages, phage e4 DNA is not circularly permuted . Adsorption under normal conditions was biphasic with rate constants of 1.02 X 10(-9)/ml/min up to 60% adsorption and 3 X 10(-10)/ml/min thereafter . Intracellular phage multiplication was characterized by a latent period of 27 min . The burst size was approximately 100 phage particles per infected cell.

J Bacteriol, 1987 May, 169(5), 2318 - 21
Characterization of Vibrio fischeri rRNA operons and subcloning of a ribosomal DNA promoter; Amikam D et al.; Analysis of rRNA genes in Vibrio fischeri indicates the presence of eight rRNA gene sets in this organism . It was found that the genes for 5S rRNA, 16S rRNA, and 23S rRNA are organized in operons in the following order: 5' end 16S rRNA 23S RNA 5S rRNA 3' end . Although the operons are homologous, they are not identical with regard to cleavage sites for various restriction endonucleases . A DNA library was constructed, and three ribosomal DNA clones were obtained . One of these clones contained an entire rRNA operon and was used as a source for subcloning . The promoter region which leads to plasmid instability was successfully subcloned into pHG165 . The terminator region was subcloned into pBR322.

Infect Immun, 1987 May, 55(5), 1090 - 3
Enterotoxicity of El Tor-like hemolysin of non-O1 Vibrio cholerae; Ichinose Y et al.; The enterotoxicity of an El Tor-like hemolysin purified from non-O1 Vibrio cholerae was investigated . Fluid accumulation was induced by injection of purified hemolysin into the ligated intestinal loops in adult rabbits (De test), intraintestinal administration in infant rabbits (Dutta test), and oral inoculation in suckling mice . The accumulated fluid was invariably mucous and bloody, and a histological change in the mucosa was observed . These results suggest that the hemolysin is an enterotoxic factor that is responsible for non-O1 V . cholerae gastroenteritis.

Infect Immun, 1987 May, 55(5), 1116 - 20
Protective efficacy in humans of killed whole-vibrio oral cholera vaccine with and without the B subunit of cholera toxin; Black RE et al.; Natural protection from cholera is associated with local intestinal antibacterial and antitoxic antibodies, which appear to act synergistically . Although current parenteral cholera vaccines offer insufficient protection, new vaccines administered orally have more promise . Killed Vibrio cholerae, alone or given with the B subunit of cholera toxin, was evaluated in adult volunteers . Vaccinees, who received three doses of either vaccine, and unvaccinated controls ingested 10(6) V . cholerae organisms to determine the protective efficacy of the vaccines . The combination vaccine provided 64% protection, and the whole vibrio vaccine given alone provided 56% protection . In addition, illnesses in vaccines were milder than those in controls, and both vaccines gave complete protection against more severe disease . This substantial level of protection against a dose of V . cholerae that caused cholera in nearly 90% of controls suggests that these vaccines might provide at least as high a level of protection if given to the population of an endemic area . Indeed, a field efficacy trial is underway in Bangladesh, and preliminary data indicate a protective efficacy of 85% for a killed whole vibrio plus B subunit vaccine similar to that tested in volunteers and an efficacy of 58% for the killed whole vibrio vaccine alone . Thus, the studies in human volunteers were successful in predicting the substantial protection afforded by the vaccines in a cholera endemic area.

J Infect Dis, 1987 May, 155(5), 979 - 84
Randomized controlled trial of berberine sulfate therapy for diarrhea due to enterotoxigenic Escherichia coli and Vibrio cholerae; Rabbani GH et al.; To evaluate the antisecretory activity of berberine sulfate (BS), we studied 165 adult patients with acute diarrhea due to enterotoxigenic Escherichia coli (ETEC) and Vibrio cholerae in randomized controlled trials . In patients with ETEC diarrhea who received 400 mg of BS in a single oral dose, the mean stool volumes were significantly less than those of the controls during three consecutive 8-hr periods after treatment (P less than .05) . At 24 hr after treatment, significantly more patients who were treated with BS and had ETEC diarrhea stopped having diarrhea as compared with the controls (42% vs 20%, P less than .05) . In patients with cholera who received 400 mg of BS, the mean 8-hr stool volume during the second 8-hr period after treatment declined to 2.22 liters, which was significantly less than the 2.79 liters found in the controls (P less than .05) . However, patients with cholera who received 1200 mg of BS plus tetracycline did not have significant reduction in stool output compared with patients who received tetracycline alone . No side effects of BS were noted . These results indicated that BS is an effective and safe antisecretory drug for ETEC diarrhea, whereas the activity against cholera is slight and not additive with tetracycline.

Gan To Kagaku Ryoho, 1987 May, 14(5 Pt 1), 1240 - 5
{Lymphocyte cytotoxicity against autologous tumor cells and the effect of interferon-beta on their activities . (1) Evaluation by modification of target tumor cells}; Ezaki K et al.; IFN is known to enhance NK activity against cultured cell lines such as K562, but not against frozen autologous tumor cells . In order to obtain increased NK cytotoxicity using IFN-beta, various modifications were performed on autologous tumor cells . IFN-beta induced more enhanced NK cytotoxicity of normal lymphocytes when frozen tumor target cells were cultured for 4-5 days in the medium, or when these cells were treated with Vibrio cholerae neuraminidase (VCN) . However, in an autologous setting, IFN-beta did not enhance NK cytotoxicity against either cultured autologous tumor cells or VCN-treated tumor cells . Also, IFN-beta did not enhance cytotoxic T cell activity against autologous tumor cells induced by mixed lymphocyte-tumor cell culture, although IFN-beta was able to induce enhancement of allospecific cytotoxic T cells mediated by mixed lymphocyte culture.

Zh Mikrobiol Epidemiol Immunobiol, 1987 May, (5), 24 - 8
{Genetic determination of the vibriocinogenicity trait in Vibrio cholerae of the El Tor biotype}; Badalova IM et al.; In two different strains of cholera vibrios two recA-dependent plasmids, pVib I (1.9-2.2 Md) and pVib II (5.2-5.8 Md), have been detected . These plasmids determine the synthesis of vibriocin, coagulase and fibrinolysin, which has been established by the cotransformation of the DNA of plasmids pVib I and pBR322 and by the transfer mobilization with the use of plasmid RP4.

Proc Natl Acad Sci U S A, 1987 May, 84(9), 2833 - 7
Use of phoA gene fusions to identify a pilus colonization factor coordinately regulated with cholera toxin; Taylor RK et al.; The transposon TnphoA was used to generate fusions between phoA, the gene for alkaline phosphatase (PhoA), and genes encoding proteins that are secreted by Vibrio cholerae . One of the PhoA+ mutants isolated showed a dramatic reduction in its ability to colonize the intestines of suckling mice . This mutant no longer produced a 20.5-kDa protein (TcpA) that we show is the major subunit of a V . cholerae pilus . Amino-terminal sequence analysis of the TcpA pilus subunit showed that it shares amino acid homology with the pilins produced by several other pathogenic bacteria . The TcpA pilus was coordinately expressed with cholera toxin under various culture conditions, and this effect appeared to be dependent on the transcriptional activator encoded by the toxR gene . We conclude that the toxR gene plays a central role in the transcriptional regulation of multiple virulence genes of V . cholerae.

J Med Microbiol, 1987 May, 23(3), 227 - 32
Studies on the Vibrio cholerae mucinase complex . II . Specific neuraminidase activity measured histochemically in a goblet cell assay; Ollar RA et al.; The activity of neuraminidase prepared from the mucinase complex of Vibrio cholerae was measured by a new, semi-quantitative goblet-cell assay . The counts of normal, alcianophilic, sialomucin-containing goblet cells (purple-stained) and neutral mucosubstance-containing goblet cells (magenta-stained) in serial sections of ileum were compared before and after neuraminidase treatment . The procedure provides a more natural assessment of the action of V . cholerae neuraminidase on the viscous intestinal mucus.

Biochem Biophys Res Commun, 1987 Apr 14, 144(1), 382 - 6
A novel mechanism for utilization of extracellular AMP in Vibrio parahaemolyticus; Sakai Y et al.; Vibrio parahaemolyticus could grow with AMP, ADP or ATP as the sole source of carbon . In the presence of Cl-, a membrane-bound Cl(-)-dependent 5'-nucleotidase seemed to hydrolyze the nucleotides extracellularly, and then the cells took up the resulting adenosine . In the absence of Cl-, although no significant dephosphorylation of the nucleotides occurred, the cells could still grow with AMP, but not with ADP or ATP . Moreover, in the presence of Cl-, Zn2+ inhibited the 5'-nucleotidase, and inhibited growth of the cells with ADP or ATP, but not with AMP, as the carbon source . V . parahaemolyticus was unable to grow with adenine or ribose 5-phosphate . These results suggested that the cells might have an AMP transport system . In fact, Na+ uptake was observed on addition of AMP to a cell suspension in the absence of Cl-, indicating Na+-AMP cotransport.

J Infect Dis, 1987 Apr, 155(4), 716 - 23
Nonlipopolysaccharide protective antigens shared by classical and El Tor biotypes of Vibrio cholerae; Sharma DP et al.; The prophylactic significance of the nonlipopolysaccharide (non-LPS) antigens of Vibrio cholerae was investigated further with use of the infant mouse cholera model . Of 16 strains examined to date, 12-including eight recent field isolates of both biotypes and both common serotypes-express common non-LPS protective antigens . The exceptional strains are four old isolates of El Tor biotype, the outer membrane proteins of which are not atypical when analyzed by immunoblotting . The protective activities of antibodies to the shared non-LPS components correlated with their capacities to inhibit the in vitro attachment of Vibrio organisms to isolated murine enterocytes.

Antibiot Med Biotekhnol, 1987 Apr, 32(4), 275 - 9
{Seasonal fluctuations in the antibiotic sensitivity of Vibrio cholerae}; Andrusenko IT et al.; Sensitivity of group 01 Vibrio cholerae to 6 antibiotics including tetracycline, ampicillin, benzylpenicillin, polymyxin M, erythromycin and novobiocin was studied during various seasons within 3 years . The antibiotic sensitivity of the cultures was assayed with the method of two-fold dilutions in solid medium AGV . The "clonal sensitivity" of the populations of the same strains was estimated by the original method developed by the authors . There were observed seasonal biorhythms in manifestation of the physiological functions by Vibrio cholerae reflected in altered "clonal sensitivity" of the populations to the antibiotics, altered multiplication intensity during various periods and seasonal dissociation with respect to the cultural and morphological features in the strains with altered properties.

Antibiot Med Biotekhnol, 1987 Apr, 32(4), 272 - 5
{Comparative antibiotic sensitivity of NAG vibrios}; Ganin VS et al.; Six hundred and forty eight NAG vibrio strains isolated at various periods from patients and carriers and from environmental objects such as surface of water reservoirs and sewage were studied with respect to their sensitivity to 14 antibiotics with the method of serial dilutions in solid media . Irrespective of the isolation place, object and time, the NAG vibrios were highly resistant to penicillins and polymyxin M . At the same time they were highly sensitive to gentamicin (MIC 1-2 micrograms/ml), levomycetin (MIC 0.5-1 micrograms/ml) and tetracyclines (MIC 0.25-1 micrograms/ml) . Study of the recipient capacity of NAG vibrios with respect to R plasmids showed that they could be recipients of exogenic R plasmids of various incompatibility groups.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1987 Apr, 264(1-2), 235 - 45
The development and application of a bacteriocinogenotyping scheme for Vibrio cholerae non-group O-1 strains; Israil AM et al.; 423 Vibrio cholerae non-group O-1 strains were tested for vibriocin production using a homologous set of 7 indicator strains . Using this scheme, 334 (79.4%) strains proved to be typable . The strains isolated from 7 different geographical areas of Romania exhibited 16 different patterns of vibriocin production . The presence of a predominant pattern per year and per region was revealed.

Infect Immun, 1987 Apr, 55(4), 942 - 6
Cell-associated hemagglutinin-deficient mutant of Vibrio cholerae; Finn TM et al.; Cell-associated hemagglutinin-negative mutants were derived from cholera enterotoxin-negative Vibrio cholerae JBK70 by Tn5 mutagenesis . One of the mutants identified, SB001, was characterized in greater detail . Its ability to colonize ilea of adult rabbits was determined by feeding approximately 10(8) V . cholerae to each animal . At 17 h after feeding, the numbers of viable vibrios in the ilea were determined . There was a significant, 4 log, decrease in the ability of the hemagglutinin-negative mutant to colonize ileal tissue compared with the parent strain JBK70 . In addition, the higher levels of colonization attained by JBK70 and the wild-type parent of JBK70, N16961, were associated with intestinal fluid accumulation and death . Rabbits immunized orally with approximately 10(8) SB001, when challenged 3 weeks later with either homologous biotype and serotype El Tor Inaba N16961 or heterologous Classical Ogawa 395, were protected to the same extent as those animals immunized with either the challenge strain or JBK70 . This was evidenced by decreases in both the number of animals showing detectable colonization and the level of colonization achieved . A hemagglutinin-negative mutant of V . cholerae may therefore be of potential use as a live oral vaccine against cholera.

FEBS Lett, 1987 Mar 9, 213(1), 81 - 4
The protein receptor for cholerabacteriophage phi 149; Ray R et al.; Choleraphage phi 149 receptor activity was found in the outer membrane (OM) protein of Vibrio cholerae 154 . Receptor protein for phage phi 149 was separated from trypsin-treated OM on a Sephadex G-100 column . Of the three peaks obtained, phage receptor activity was noted only in peak II . SDS-PAGE showed that the Mr of the protein was 35,000 . The protein was heat-labile and protease-sensitive . The specificity of this protein as choleraphage phi 149 receptor was investigated by carrying out a protection experiment by anti-protein (peak II) rabbit sera.

Eur J Biochem, 1987 Mar 2, 163(2), 407 - 16
Structural characterization of gangliosides from murine T lymphocytes; Muthing J et al.; Mouse spleen cells were prepared from CBA/J mice, and T lymphocytes were selectively stimulated with the T cell mitogen concanavalin A and further propagated in the presence of the T cell growth factor interleukin-2 . The T cells were metabolically labeled with D-{1-14C}galactose and D{1-14C}glucosamine, and the gangliosides were extracted and purified by DEAE-Sepharose column chromatography . Carbohydrate backbone structures of the asialogangliosides, prepared by mild acid hydrolysis, were determined by high-performance liquid chromatography, treatment with exoglycosidases and immunostaining . Monosialylated gangliosides were isolated by gradient elution from DEAE-Sepharose and further separated by preparative high-performance thin-layer chromatography in two solvent systems . Isolated fractions were characterized by preparation of asialogangliosides by mild acid hydrolysis, the action of Vibrio cholerae neuraminidase, and fast-atombombardment mass spectrometry . The following structures were identified: IVNeuAc-GgOse4Cer; IVNeuGc-GgOse4Cer; IVNeuAc-GgOse5Cer; and IVNeu-Gc-GgOse5Cer . The latter two gangliosides were not detected on B lymphoblasts and may be T-cell-specific structures . All gangliosides were heterogeneous in their ceramide moieties, being substituted with C16:0, C24:0, and C24:1 fatty acids . A preliminary study of several other mouse strains showed no strain-specific genetic variations in the T cell gangliosides . The possible role of these gangliosides is discussed.

Am J Trop Med Hyg, 1987 Mar, 36(2), 393 - 7
Non-01 Vibrio cholerae infections in Cancun, Mexico; Finch MJ et al.; To determine the role of Vibrio cholerae as a cause of diarrheal illness in Cancun, Mexico, an investigation was conducted in July and August 1983 . Although toxigenic V . cholerae 01 were not found, non-01 V . cholerae were isolated from 22 (16%) of 134 stools from persons with diarrheal illness and none of 22 stools from well persons; 58 (92%) of 63 sewage samples; 12 (86%) of 14 untreated well water samples; a home storage tank for treated water; and 5 (21%) of 24 samples of raw seafood . None of the V . cholerae isolates from patients were toxigenic . The illness occurred mainly in small children, and were characterized principally by diarrhea and abdominal pain . No patient was seriously ill, and all recovered without sequelae . Seven different serotypes of non-01 V . cholerae were isolated from the stool specimens, and Smith serotype 12 accounted for 10 (46%) of the 22 isolates . A matched-pair case-control study found that cases were more likely than controls to have eaten home prepared gelatin (P = 0.03, OR = 5/0) and seafood (P = 0.06, OR = 4/0).

Gastroenterology, 1987 Mar, 92(3), 796 - 9
Vibrio vulnificus infection after raw oyster ingestion in a patient with liver disease and acquired immune deficiency syndrome-related complex; Chin KP et al.; Sepsis, peritonitis, and gastroenteritis developed in a 45-yr-old homosexual man 1 day after ingestion of raw oysters . The patient had chronic active hepatitis and cirrhosis with hepatitis B virus and delta-infection . He also had persistent generalized lymphadenopathy associated with HTLV-III antibody positivity . Vibrio vulnificus was isolated from the patient's blood and peritoneal fluid as well as from the same batch of oysters at the restaurant where the patient had visited . To our knowledge, this is the first report relating direct microbiologic and clinical evidence that the infection is acquired through the gastrointestinal tract by consuming raw seafood containing the pathogen . This is also the first reported case of peritonitis associated with sepsis and gastroenteritis from this organism . Patients with liver disease and other immunocompromised states should be warned about such life-threatening infections and complications associated with the consumption of raw oysters or other undercooked seafoods.

Southeast Asian J Trop Med Public Health, 1987 Mar, 18(1), 33 - 8
Antibodies against Vibrio cholerae lipopolysaccharide, cell-bound haemagglutinin and toxin in the intestinal fluid during convalescence; Chongsa-Nguan M et al.; Specific antibodies to V . cholerae lipopolysaccharide (LPS), cell-bound haemagglutinin (CHA) and toxin (CT) in the intestinal lavages of healthy Thais and Thai cholera patients during the convalescence period were determined by enzyme-linked immunosorbent assay . Only IgM and IgA specific antibodies were detectable in the specimens . All of the persons who were just recovered from cholera had IgA anti-CT and IgA anti-LPS and 82.4% had IgA anti-CHA . The IgA anti-CT, anti-LPS and anti-CHA were detected also in the gut fluids of 70.6%, 94.1% and 88.2%, respectively, of the healthy controls . The mean levels of the IgA antibodies of all specificities between the two groups of individuals were not different . However, the IgM anti-CT and IgM anti-LPS of the cholera patients increased during the convalescence period . The levels, therefore, were significantly higher than those of the controls . The ratios of IgA anti-CT: IgM anti-CT and IgA anti-LPS: IgM anti-LPS among the patients were 2.93:1 and 2.02:1, respectively while those of the controls were 10:1 and 34:1, respectively . IgA antibodies predominated in the lavages of both groups of the individuals.

Ann Inst Pasteur Microbiol, 1987 Mar-Apr, 138(2), 223 - 34
{Assay of thermolabile enterotoxins of Escherichia coli and Vibrio cholerae by inhibition of erythroadsorption on the GM1 ganglioside}; Germani Y et al.; A GM1 erythroassay (GERYDO) for heat-labile Escherichia coli enterotoxin (LT) and cholera toxin (CT) is described . This assay was developed for use in poorly equipped laboratories in developing countries . It uses GM1-coated polystyrene plates and is based on the competition between the toxin to be assayed and CT covalently bound to sheep red blood cells . GERYDO can detect 0.9 or 0.5 ng of CT per ml depending on the method of sensitization of erythrocytes . Good quantitative and qualitative correlation with the enzyme-linked immunosorbent assay and the Vero cell test was observed.

J Appl Bacteriol, 1987 Mar, 62(3), 279 - 87
Ampicillin-dextrin agar medium for the enumeration of Aeromonas species in water by membrane filtration; Havelaar AH et al.; Published selective media were evaluated for the isolation of Aeromonas spp . from environmental samples by membrane filtration . Satisfactory recoveries were obtained only with mA agar (Rippey & Cabelli) and dextrin-fuchsin-sulphite agar (Schubert), but neither was sufficiently selective . The positive aspects of these two media were combined in a new medium, ampicillin-dextrin agar . Recovery from pure cultures and environmental samples was optimal at an ampicillin concentration of 10 mg/l and incubation for 24 h at 30 degrees C under aerobic conditions, and specificity was high (i.e . confirmation rate usually greater than 90%, no false negative colonies encountered) . The medium can also be used for isolation of Aeromonas spp . from sea water provided that the vibriostatic agent 0/129 is added at 50 mg/l.

Appl Environ Microbiol, 1987 Mar, 53(3), 603 - 5
Effects of chitin and its soluble derivatives on survival of Vibrio cholerae O1 at low temperature; Amako K et al.; Chitin concentrations greater than 0.04% (wt/wt) protected cholera vibrios against killing at low temperature . This protective effect was detected with both the soluble form of chitin, glycol chitin, and the insoluble particulate form of chitin . Some amino acids or peptides also showed the same protective effect.

Br J Haematol, 1987 Mar, 65(3), 351 - 5
Decreased sialic acid content of erythrocytes in patients with aplastic anaemia; Yoshida M et al.; The sialic acid content of erythrocytes from patients with aplastic anaemia was determined and compared with those in patients with several haematological disorders and healthy individuals . The sialic acid was released enzymatically with Vibrio cholerae sialidase and quantitated by the thiobarbituric acid method (Aminoff, 1961) . The sialic acid content of normal erythrocytes was 538 +/- 31 nmol/ml of packed erythrocytes . That of erythrocytes from patients with aplastic anaemia was 480 +/- 35 nmol/ml of packed erythrocytes, which was significantly lower than normal (P less than 0.01) . In contrast, erythrocytes from patients with myeloproliferative disorders showed significantly (P less than 0.05) higher sialic acid contents (564 +/- 45 nmol/ml of packed erythrocytes) . These results suggest that some membrane changes occur in erythrocytes in patients with these disorders.

J Bacteriol, 1987 Mar, 169(3), 1352 - 7
Evolutionary origin of pathogenic determinants in enterotoxigenic Escherichia coli and Vibrio cholerae O1; Yamamoto T et al.; Three families of the evolutionarily related pathogenic determinants in enterotoxigenic Escherichia coli and Vibrio cholerae O1, a family of cholera enterotoxin (CT) and heat-labile enterotoxin (LT) including CT, LTh, and LTp, a family of heat-stable enterotoxin I (STI) including STIa and STIb, and a family of K88 enteroadhesion fimbriae including K88ab, K88ac, and K88ad were analyzed for synonymous (silent) nucleotide substitutions by using the gene nucleotide sequences of earlier reports and the LTp gene nucleotide sequence presented in this paper . The data suggested that the divergences between LT and CT and between STIa and STIb occurred in the remote past, whereas those between LTh and LTp and between members of the K88 family occurred very recently . We concluded that the LT gene is a foreign gene that has been acquired by E . coli to form an enteropathogen . This provides evolutionary evidence of species-to-species transfer of pathogenic determinants in procaryotes.

J Bacteriol, 1987 Mar, 169(3), 1037 - 45
Transient entry of enterotoxin subunits into the periplasm occurs during their secretion from Vibrio cholerae; Hirst TR et al.; Cholera toxin and heat-labile enterotoxin (LT) are structurally similar oligomeric proteins which are capable of being efficiently secreted from Vibrio cholerae . Here we report that these proteins transiently enter the periplasm of V . cholerae as they traverse the cell envelope to reach the extracellular milieu . Pulse-chase experiments on V . cholerae TRH7000 harboring an LT-encoding plasmid revealed that radiolabeled LT A and B subunits entered the periplasm rapidly, followed by their slow efflux (half-time, 13 min) into the medium . LT B-subunit efflux from the periplasm was calculated to be at a rate of ca . 170 monomers per min per cell (which is equivalent to 34 assembled LT holotoxin molecules per min per cell) . These values were estimated to be sufficient to account for the increase in extracellular enterotoxin concentration during exponential cell growth . Thus, all enterotoxin subunits which are secreted into the medium can be assumed to be channelled via the periplasm . These findings led to an improved model of the pathway of toxin secretion by V . cholerae.

Proc Natl Acad Sci U S A, 1987 Mar, 84(6), 1709 - 12
Role of sialic acid in synaptosomal transport of amino acid transmitters; Zaleska MM et al.; Active, high-affinity, sodium-dependent uptake of gamma-aminobutyric acid and of the acidic amino acid D-aspartate was inhibited by pretreatment of synaptosomes with neuraminidase from Vibrio cholerae . Inhibition was of a noncompetitive type and was related to the amount of sialic acid released . The maximum accumulation ratios of both amino acids (intracellular {amino acid}/extracellular {amino acid}) remained largely unaltered . Treatment with neuraminidase affected neither the synaptosomal energy levels nor the concentration of internal potassium . It is suggested that the gamma-aminobutyric acid and acidic amino acid transporters are glycosylated and that sialic acid is involved in the operation of the carrier proteins directly and not through modification of driving forces responsible for amino acid uptake.

J Med Microbiol, 1987 Feb, 23(1), 9 - 18
Preparation and immunogenicity of a bivalent cell-surface protein-polysaccharide conjugate of Vibrio cholerae; Kabir S; Alkali-treated lipopolysaccharides (LPS) from Ogawa and Inaba serotypes of Vibrio cholerae were chemically coupled to cell-surface proteins of V . cholerae . The reaction product was eluted in the void volume when fractionated on a column of Sephacryl S-300 . The material did not enter the gel when subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate (SDS-PAGE) . The bivalent protein-polysaccharide conjugate was nonpyrogenic, as determined by the Limulus lysate assay . It was immunogenic and elicited, in rabbits, antibodies against both intact LPS and cell surface proteins, as determined by enzyme linked immunosorbent assay . LPS from Ogawa serotype was resolved into two major bands by SDS-PAGE and that from the Inaba serotype into one major band . Immunoblotting studies indicated that antisera to the protein-polysaccharide conjugate contained antibodies to the major LPS fractions from both serotypes . Antisera to the protein-polysaccharide conjugate tested by crossed-immunoelectrophoresis produced immunoprecipitation with whole-cell sonicates of both biotypes and serotypes of V . cholerae . Such antisera also possessed agglutinating and complement-mediated bactericidal activities towards V . cholerae strains of both biotypes and serotypes . These results suggest that a bivalent cell-surface protein-polysaccharide conjugate of V . cholerae could be developed as a nonpyrogenic vaccine against cholera.

Diagn Microbiol Infect Dis, 1987 Feb, 6(2), 109 - 17
Clinical and ecological characteristics of Vibrio vulnificus in the northeastern United States; Tilton RC et al.; Multiple seawater sites in the northeastern United States, particularly Long Island Sound, and shellfish from Long Island Sound were sampled from April to November for 3 successive yr, 1983-1985 . Hospitals in coastal and metropolitan areas of Connecticut were surveyed for the same 3-yr period, Vibrio vulnificus can be found in these waters during the summer months . The appearance of these virulent bacteria in both seawater and shellfish are a function of the water temperature; no V . vulnificus could be isolated until the temperature was approximately 17 degrees C . Although the risk of infection is small, as shown by isolation of this organism from patients, certain high-risk groups exist . Consumption of raw shell fish during the summer months should be discouraged in people with liver disease or patients on immunosuppressive therapy.

J Infect Dis, 1987 Feb, 155(2), 236 - 41
Mouse skin damage caused by cytolysin from Vibrio vulnificus and by V . vulnificus infection; Gray LD et al.; Light and electron microscopy of mouse skin damage caused by intradermal infection with a virulent strain of Vibrio vulnificus and by a single intradermal injection of the cytolytic toxin produced by the bacterium revealed similar structural alterations . The epidermis was intact; however, the infection and toxin produced acute cellulitis characterized by extensive extracellular edema; disorganization of collagen bundles; large accumulations of cell debris and plasma proteins; damaged or necrotic fat cells, capillary endothelial cells, and muscle cells; and mild inflammatory cell infiltration . The virulent strain of V . vulnificus produced a capsule and was resistant to phagocytosis in vivo, whereas a weakly virulent strain of the bacterium did not produce a capsule and was readily phagocytized and digested . Factors that may be important in the pathogenesis of V . vulnificus wound infections include a capsule that inhibits phagocytosis and an extracellular cytolytic toxin that is responsible, at least in part, for the severe tissue damage characteristic of such wound infections.

Infect Immun, 1987 Feb, 55(2), 477 - 81
Determinants of the immunogenicity of live virulent and mutant Vibrio cholerae O1 in rabbit intestine; Pierce NF et al.; Determinants of the capacity of live Vibrio cholerae O1 isolates to evoke specific immune responses in intestinal mucosa were studied in rabbits, using mucosal immunoglobulin A (IgA) antitoxin as the measured immune response . Antitoxin responses were evoked mostly by the primary inoculation and were dose dependent; secondary-type responses were modest and occurred only when the booster inoculum was large, i.e., 10(10) CFU . The efficiency of mucosal immunization correlated closely with the mucosal colonizing capacity of the infecting strain and was otherwise independent of toxin genotype (A+ B+ or A- B+) or whether the strain was motile or nonmotile . Live bacteria evoked mucosal antitoxin more efficiently than did purified cholera toxin . Prior immunization with a nontoxinogenic (A- B-) V . cholera strain interfered significantly with the induction of mucosal antitoxin by subsequent immunization with its fully toxinogenic (A+ B+) parent . These results demonstrate the marked efficiency with which live V . cholerae stimulate a specific enteric mucosal secretory IgA response . They support the view that mucosal colonization aids efficient delivery of bacterial antigens to responsive subepithelial lymphoid tissue . This might occur by transfer of colonizing bacteria through M cells into Peyer patches or by efficient delivery of secreted toxin to M cells by mucosa-associated organisms . Preexisting antibacterial immunity interferes with colonization, which may prevent efficient antigenic stimulation and which may explain the relatively weak response to booster immunization . The same process may also limit the efficacy of hybrid enteric bacterial vaccines when there is preexisting mucosal immunity to the carrier organism due to either natural exposure or prior immunization with another vaccine that uses the same carrier.

Zh Mikrobiol Epidemiol Immunobiol, 1987 Feb, (2), 15 - 8
{Lecithinase activity of Vibrio cholerae and of vibrios not agglutinated by O-cholera serum}; Nabiev EG et al.; The lecithinase activities of 187 V . cholerae strains differing in their virulence and 70 cultures of NAG vibrios of different origin varied in all experimental groups . The level of lecithinase activity in the groups of V . cholerae strains having low virulence, or no virulence at all, was considerably higher than in virulent strains . NAG vibrios isolated from diarrhea patients and from samples of lake and pond water did not differ in the activity of this enzyme.

Infect Immun, 1987 Feb, 55(2), 451 - 4
Comparative study of Vibrio cholerae non-O1 protease and soluble hemagglutinin with those of Vibrio cholerae O1; Honda T et al.; Protease and soluble hemagglutinating activities produced by a non-O1 Vibrio cholerae strain isolated from a patient with diarrhea were compared with similar activities produced by V . cholerae O1 . The soluble protease activities were indistinguishable in heat stability, immunodiffusion, inhibition by antiserum, and electrophoretic analysis . On the other hand, the soluble hemagglutinating activities of both strains were not completely identical . The hemagglutinating activity of the non-O1 V . cholerae strain was not inhibited by Zincov; it was more sensitive to inhibition by normal serum, and it had an unusual pattern of heat stability . Heating at 100 degrees C resulted in some recovery of activity of a sample previously inactivated by heating at 60 degrees C.

Appl Environ Microbiol, 1987 Feb, 53(2), 466 - 9
Salt tolerance of lactose-grown Vibrio parahaemolyticus carrying Escherichia coli lac genes; Datta AR et al.; Lac- strains of Vibrio parahaemolyticus were converted to Lac+ on receiving a hybrid plasmid containing the lactose utilization genes of Escherichia coli K-12 . A V . parahaemolyticus strain containing this hybrid plasmid exhibited optimal growth rates on glucose and other carbon sources in the presence of 0.2 to 0.4 M NaCl . Growth of the same strain on lactose was inhibited at similar concentrations of NaCl . The altered growth rate responses in lactose medium appeared to be attributable to effects of NaCl on the activity of lactose permease, and possibly on that of beta-galactosidase, rather than on the levels of these enzymes in V . parahaemolyticus cells.

Life Sci, 1987 Jan 5, 40(1), 55 - 62
Trace amounts of ganglioside GM1 in human milk inhibit enterotoxins from Vibrio cholerae and Escherichia coli; Laegreid A et al.; Gangliosides were isolated from human milk fat and purified by silica gel column chromatography and high performance liquid chromatography (HPLC) . Low amounts of the ganglioside GM1, detected by high performance thin layer chromatography (HPTLC)-immunoassay, were found in all fractions with enterotoxin-inhibitory activity, while fractions without GM1 were inactive . It is concluded that GM1 is responsible for enterotoxin-inhibitory activity in the ganglioside fraction from human milk.

Comp Biochem Physiol B, 1987, 86(1), 31 - 6
Effect of opsonization on oxidative metabolism of plaice (Pleuronectes platessa L.) neutrophils; Nash KA et al.; The effect of serum opsonization on Vibrio alginolyticus (heat-killed)-stimulated chemiluminescence (CL) by plaice kidney- and peritoneal exudate-derived neutrophils was investigated . Peritoneal neutrophils only recognized heat-labile and kidney neutrophils only heat-stable opsonic activity in normal serum . Specific antibody did not show opsonic activity nor any synergism with the normal serum opsonins for either neutrophil population . Evidence was found for the production, by plaice neutrophils, of H2O2, O2-, OH . and two or more, as yet unidentified, reactive oxygen species (ROS).

Appl Environ Microbiol, 1987 Jan, 53(1), 193 - 5
Identification of environmental Vibrio vulnificus isolates with a DNA probe for the cytotoxin-hemolysin gene; Morris JG Jr et al.; We screened 44 lactose-positive Vibrio strains isolated from the marine environment for homology with a 3.2-kilobase DNA fragment encoding the Vibrio vulnificus cytotoxin-hemolysin gene . All 29 marine isolates identified as V . vulnificus on the basis of numerical taxonomy and DNA-DNA hybridization studies hybridized with the cytotoxin gene probe, as did all V . vulnificus reference strains . Homologous gene sequences were identified in no other lactose-positive marine vibrio isolates nor in 10 other Vibrio species.

Biokhimiia, 1987 Jan, 52(1), 15 - 23
{The role of Na+ ions in the respiration, formation of the membrane potential and movement of the alkali-resistant marine bacterium Vibrio alginolyticus}; Dibrov PA et al.; Subbacterial vesicles capable of generating delta psi during NADH oxidation were obtained . The oxidation of NADH was stimulated by Na+ and inhibited by 2-heptyl-4-oxyquinoline-N-oxide (HQNO) in submicromolar concentrations . The generation of delta psi was inhibited by HQNO in low concentrations, cyanide, gramicidine D and carbonyl cyanide-m-chlorophenylhydrazone (CCCP) in combination with monensine . At the same time, in the absence of monensine CCCP influenced the delta psi generation in a much lesser degree . In subbacterial vesicles delta psi generation coupled with NADH oxidation necessitated Na+ . Experiments with intact cells of V . alginolyticus revealed that cell motility depends on Na+, is sensitive to CCCP + monensine as well as to arsenate + HQNO, cyanide or anaerobiosis . In the absence of arsenate, the inhibition of respiration partly decreased the rate of bacterial movement . In the presence of HQNO and arsenate, NaCl addition to K+-loaded cells led to the monensine preventing restoration of the cell motility during a few minutes . However, no stimulating effect was observed in the case of artificial delta pH formation as a result of acidification of the medium (from pH 8.6 to pH 6.5) . The experimental results suggest that delta mu Na+ generated by the respiratory chain and by the arsenate-sensitive enzymatic system (presumably, glycolysis and Na+-ATPase) can be utilized by the Na+-driven molecular motor responsible for the motility of V . alginolyticus cells.

Strahlenther Onkol, 1987 Jan, 163(1), 43 - 9
{Effect of neuraminidase and x-rays (2 Gy and 8 Gy) on microvilli and membrane invaginations of Ehrlich ascites tumor cells in monolayer culture}; Laudenbach G et al.; A monolayer culture (Eagle basal medium plus 10% of fetal calf serum) of Ehrlich ascites tumor cells was exposed to X-radiation with 2 Gy and 8 Gy and treated with Vibrio cholerae neuraminidase alone or combined with sublethal X-ray irradiation (2 Gy) . Pictures of the Ehrlich ascites tumor cells taken with the electron microscope were investigated in order to find out any cell surface modifications due to membrane invaginations and microvilli . The results showed that the rate of microvilli as well as that of membrane invaginations became higher with the increasing X-ray dose (2 Gy; 8 Gy) . Following to neuraminidase treatment there was a considerable augmentation of membrane invaginations as compared to control cells, whereas the number of microvilli was slightly reduced . As it has been already described before, the influence of neuraminidase produced an increased endocytosis activity and a strengthening of the cytoskeleton . Combined treatment with neuraminidase and sublethal X-radiation (2 Gy) caused a higher rate of membrane invaginations than each method alone; the number of microvilli was slightly increased by combined treatment . The conclusion is drawn that these structure modifications are due to reparation processes induced by radiation on the one hand and to an enzymic action of neuraminidase on the cell surface on the other hand.

Microbiol Immunol, 1987, 31(8), 763 - 70
Oral immunization in adult mice to live and heat-killed Vibrio cholerae; Fujisawa H et al.; Effects of systemic and intestinal local immune responses in mice fed ad libitum and forcedly with live or heat-killed Vibrio cholerae on the elimination of vibrios from the intestine were investigated . In mice fed with live vibrios, ad libitum feeding could induce potential delayed-type hypersensitivity and rapid production of vibriocidal antibody in the serum whereas forcedly feeding suppressed the delayed-type hypersensitivity and retarded the antibody production . In contrast, when killed microorganisms were used as antigens, significant delayed-type hypersensitivity and rapid response of vibriocidal antibody were induced in forcedly fed mice although ad libitum feeding suppressed the induction of the delayed-type hypersensitivity and retarded the production of vibriocidal antibody . The elimination of vibrios from the intestine of mice was promoted in both mice groups fed ad libitum and forcedly with live vibrios but not with killed microorganisms . Total IgA in the intestinal contents of mice fed with live vibrios both ad libitum and forcedly were higher than those of mice fed with killed antigens . In addition, when the extracts of intestinal contents were absorbed by live antigens, IgA contents in mice fed with live vibrios were reduced more markedly than those in mice immunized orally by the feeding with killed antigens . These findings suggested that the elimination of vibrios from the organ was closely related to local IgA antibody response to heat-labile substance of live Vibrio cholerae.

Microbiol Immunol, 1987, 31(8), 753 - 61
Quantitative evaluation of colonizing ability of Vibrio cholerae O1; Nakasone N et al.; A new method to evaluate the adhesive ability of Vibrio cholerae O1 was proposed . Broth cultured V . cholerae O1 and a piece of formalin-fixed rabbit intestinal wall were incubated together in KRT buffer and the number of adhered organisms was counted under a scanning electron microscope . This method was much less laborious than other methods that have been used so far, and most significantly, constant results were obtained in repeated experiments . The adhesive properties of toxigenic V . cholerae O1 evaluated by this method correlated well with its observed experimental pathogenicity.

Dev Comp Immunol, 1987 Summer, 11(3), 529 - 37
The specificity of Atlantic salmon antibodies made against the fish pathogen Vibrio salmonicida, establishing the surface protein VS-P1 as the dominating antigen; Espelid S et al.; The specificity of salmon (Salmo salar) antibodies made against the fish pathogen Vibrio salmonicida was studied . Salmon immunized with V . salmonicida emulisified in Freund's complete adjuvant produced antibodies which preferentially bound to a 40 KD outer surface molecule (VS-P1) . Moreover, the bulk of the antibodies were specific for one particular epitope on VS-P1, defined by a mouse monoclonal antibody - as detected with a blocking ELISA . The data imply that salmon B cells mainly (90%) recognize one particular determinant on V . salmonicida and thus express a limited repertoire of antibody specificities against this pathogen.

Trans R Soc Trop Med Hyg, 1987, 81(2), 230 - 2
Epidemiological characteristics of an institutional outbreak of cholera; Goh KT et al.; An outbreak of cholera caused by Vibrio cholerae 01, biotype El Tor, serotype Inaba, phage type 4, occurred in an institution for the aged in Singapore in August and September 1984 . 96 inmates were infected (21 symptomatic and 75 asymptomatic) and 5 died . The index case was a 72-year-old male inmate who continued to assist in food preparation in the kitchen from the time of onset of diarrhoea until he was seriously ill and hospitalized 4 days later . Another kitchen helper was found to have asymptomatic V . cholerae 01 infection . The infection rate for males was significantly higher than that for females (P less than 0.025), associated with the use of unsanitary toilets . The main mode of transmission was through food contaminated by the 2 kitchen helpers who probably accounted for most of the infections, while poor personal hygiene of the inmates helped to sustain person-to-person spread . The outbreak was confined within the institution as the result of the prompt and effective implementation of control measures.

Trop Geogr Med, 1987 Jan, 39(1), 9 - 14
Breast-feeding children in the household as a risk factor for cholera in rural Bangladesh: an hypothesis; Riley LW et al.; Vibrio cholerae 01 produces symptomatic and asymptomatic infections . In this study, we investigated a cholera epidemic in northern Bangladesh to specifically search for risks of developing symptomatic infection . A case-control study in six villages found that cases were more likely than controls to have in their family a child who was still breast-feeding and who had been asymptomatic during the epidemic . Among 24 case-control pairs with cholera-like diarrhea as cases, there were 11 discordant for the presence of such a child in the family, in 9 of them, the child was in the case-family (relative risk = 4.5, p = 0.033) . Among 13 case-control pairs with laboratory-confirmed cholera as cases, there were 7 discordant for the presence of a breast-feeding child, and in 6 of them, the child was in the case-family (relative risk = 6, p = 0.06) . Breast-feeding children in this area are usually kept naked, and defecate onto a cloth pad held against their buttocks by a family member who may be repeatedly exposed to the soiled cloth . Symptomatic infection with V . cholerae may depend on exposures to situations that augment the ingested dose of V . cholerae, and these findings led us to hypothesize that breast-feeding children, if infected, may play a substantial role (attributable risk = 55%) in facilitating such transmission in rural Bangladesh.

Eksp Onkol, 1987, 9(2), 36 - 40
{Modulating effect of immunotherapy with modified tumor cells in transplantable lymphoma}; Bratus' GG et al.; Vibrio cholerici neuraminidase was studied on CBA mice for its effect on the strain NK/Ly lymphoid cells oncogenicity in intraabdominal transplantation . The 30% decrease in transplantability of cells treated by the enzyme in the dose of 50 U/ml was established . The modulating effect of specific active immunotherapy by modified cells in lymphoma was also studied . A pronounced stimulation of immunity T-cell links and proliferation in immunocompetent organs, an increase in the number of large granular lymphocytes in blood, specific antibody formation suppression were established . The problem of neuraminidase nonspecific effect on the tumour cells is under discussion.

Ann Rech Vet, 1987, 18(1), 43 - 6
In vitro and in vivo study of an antimicrobial activity displayed by the redmouth disease agent, Yersinia ruckeri; Michel C et al.; A soluble antimicrobial product has been detected in Yersinia ruckeri cultures . The contract droplets technique and the double agar layer method allowed to assess its bacteriostatic effect and the high susceptibility of other bacterial fish pathogens like Vibrio and Aeromonas . But it did not inhibit other strains of Y ruckeri . No correlation was found between the antimicrobial activity and the virulence of the strains . As mixed infections with A salmonicida and Y ruckeri could be obtained experimentally, the factor does not seem to confer any competitive advantage in fish tissues colonization.

Biochem Cell Biol, 1987 Jan, 65(1), 19 - 26
Structural elucidation of the O-specific polysaccharide of the phenol-phase soluble lipopolysaccharide of Vibrio anguillarum; Banoub JH et al.; The structure of the O-specific polysaccharide of the phenol-soluble cellular lipopolysaccharide of Vibrio anguillarum has been investigated . The studies involved the use of methylation analysis, partial hydrolysis with 48% hydrogen fluoride, Smith degradation, oxidation with chromium trioxide, and comprehensive proton and carbon-13 nuclear magnetic resonance studies, in which one- and two-dimensional experiments were carried out . As a result of these studies it is proposed that the O-specific polysaccharide of Vibrio anguillarum is composed of a regular heteropolymer, i.e., a main chain of (1----4)-linked 3-acetamido-3,6-dideoxy-beta-L-glucose residues alternately substituted through O-2 with side chain residues of 2-acetamido-2,6-dideoxy-alpha-D-glucose, which seem to be substituted through either O-3 or O-4 with propionyl groups (R), as in the following structure . (Formula: see text)

Med Microbiol Immunol (Berl), 1987, 176(2), 89 - 97
Assay for heat-labile Escherichia coli enterotoxin using sandwich erythroimmunoassay; Germani Y et al.; We investigated the possibility of using a sheep erythrocyte-antibody conjugate as reagent in a sandwich erythroimmunoassay (SERIA) procedure to detect and titrate Escherichia coli heat-labile enterotoxin (LT) with the naked eye . In this assay, which is based on the immunological similarity between Vibrio cholerae toxin (CT) and LT, rabbit anti-CT IgG was used as immunosorbent, and sheep erythrocytes, sensitized with the rabbit anti-CT antibodies, were used as indicator . The sensitivity of the test was demonstrated by a comparative study using an enzyme-linked immunosorbent assay (ELISA) . The results obtained by SERIA with 130 samples correlated well with those of a Vero cell assay and a GM1-ELISA . The test is easy, relatively cheap and as sensitive as other standard techniques; it is particularly suited for field laboratories, especially in tropical countries, and a large number of strains may be examined daily.

J Infect Dis, 1987 Jan, 155(1), 79 - 85
B subunit-whole cell and whole cell-only oral vaccines against cholera: studies on reactogenicity and immunogenicity; Clemens JD et al.; We conducted a randomized trial among persons in rural Bangladesh to evaluate the side effects and immunogenicity of orally administered B subunit-killed whole cell (BS-WC) and killed whole cell-only (WC) cholera vaccines and a killed Escherichia coli strain K12 placebo proposed for field testing . Three doses of BS-WC, WC, E . coli, or a control agent were given with antacid to 1,257 women (aged greater than 15 years) and children (aged to to 15 years) . The four groups exhibited no statistically significant differences in occurrence of symptoms after each dose, and rises in titers of vibriocidal (VC) antibodies to Inaba and Ogawa were twofold higher for vaccinees than for controls (P less than .001) . Half of the persons with fourfold or greater VC responses to WC responded after the first dose; many additional patients, particularly young children, responded after subsequent doses . In contrast, 89% of persons who responded to BS-WC with twofold or greater rises in titer of IgG antibodies to cholera toxin did so after the first dose . After the third dose, vaccinees exhibited a fivefold higher rise in titer than did controls (P less than .001); a dose-to-dose booster effect was most evident in young children.

J Bacteriol, 1987 Jan, 169(1), 247 - 53
Expression of bioluminescence by Escherichia coli containing recombinant Vibrio harveyi DNA; Miyamoto C et al.; When isogenic strains of Escherichia coli, RR1 (rec+) and HB101 (recA), were transformed with mapped recombinant plasmids known to contain Vibrio harveyi luciferase genes and large regions of DNA flanking on both sides, a small percentage (0.005%) of the colonies expressed high levels of luminescence (up to 10(12) quanta s-1 ml-1) in the absence of added aldehyde . The altered ability to express light was found to be due to a mutation in the host and not to an alteration in the recombinant DNA . When these bright colonies were cured of plasmid, they could be retransformed with cloned V . harveyi gene fragments in cis and in trans to yield luminescent colonies at 100% frequency . The maximum length of V . harveyi DNA required to produce light-emitting E . coli was shorter (6.3 kilobase pairs) than that required for expression of the V . fischeri system in E . coli . Cell extracts from bright clones contained wild-type levels of activity for the heteropolymeric (alpha beta) luciferase; fatty acid labeling revealed the presence of the three acylated polypeptides of the fatty acid reductase system which is involved in aldehyde biosynthesis for the luminescence reaction . The increased light emission in the mutant bacteria appeared to arise in part from production of higher levels of polycistronic mRNAs coding for luciferase.

Trans R Soc Trop Med Hyg, 1987, 81(5), 876 - 8
Characterization of Vibrio cholerae 01 recently isolated in Bangladesh; Nakasone N et al.; 91 strains of Vibrio cholerae O1, isolated in Bangladesh in January 1986, were examined for their biological behaviour and sensitivity to 6 antimicrobial agents . Biotyping indicated that 60 of the isolates belonged to the classical biotype and 31 to the El Tor biotype . 21 El Tor strains revealed beta-haemolysis on blood agar plates, but only 8 showed complete haemolysis in broth . Serotyping indicated 79 Ogawa, 10 Inaba, and 2 Hikojima . Phage typing showed that all classical vibrios belonged to Mukerjee's phage type 1 . El Tor vibrios were classified into 4 groups: one strain each in type 1 and type 5, 19 in type 4, and 10 in an untypable group . Prophage typing of El Tor vibrios identified 14 strains of Ubol type, 16 of cured Celebes type, and one of original Celebes type . No strain was resistant to tetracycline, minocycline, chloramphenicol, streptomycin, amoxicillin or nalidixic acid . The classical vibrios differed from those isolated before 1973 in toxin production pattern.

Int J Immunopharmacol, 1987, 9(7), 841 - 50
Tumor therapy of neoplastic diseases with tumor cells and neuraminidase: experimental studies on chessboard vaccination in transplantation tumors; Sedlacek HH et al.; The therapeutic effect of intradermal (i.d.) injection of tumor cells mixed with VCN on growth of spontaneous metastases in transplantable tumors in mice (3-Lewis lung adenocarcinoma; B16-melanoma) and rats (R-3230 mammary adenocarcinoma) was investigated . Intradermal injection was done in a chessboard-like manner; increasing numbers (10(5), 10(6) and 10(7)) of Mitomycin-treated tumor cells (M-TC) were each mixed with increasing amounts (10, 50 and 100 mU) of Vibrio cholerae Neuraminidase (VCN) . These different mixtures were injected i.d . at different sites one day after resection of the primary tumor graft to mice and rats, suffering from minimal residual disease . The therapeutic effect of this so-called chessboard vaccination on minimal residual disease was compared to that of the subcutaneous or i.d . injection of VCN-treated M-TC (10(5), 10(6), 10(7) or 10(8) cells) or of single mixtures of M-TC and VCN . The results show that compared to VCN-treated M-TC or single mixtures of M-TC and VCN, chessboard vaccination is the only procedure that is therapeutically effective on metastasation of Lewis lung adenocarcinoma . The therapeutic effect could be abrogated by heat-inactivation of VCN . Incomplete chessboard vaccinations (10(5), 10(6), 10(7) tumor cells, each mixed with 5 mU VCN only) were likewise ineffective . However, treatment with incomplete chessboard vaccinations in combination with a low dose of cyclophosphamide (which is not immunosuppressive, but partly inhibits tumor growth) had a synergistic therapeutic effect on minimal residual disease of Lewis lung adenocarcinoma . In contrast, growth of metastases of B16-melanoma and R-3230 adenocarcinoma could not significantly be influenced by any of those treatments . The DTH response of tumor bearing animals against i.d . applied tumor cells was neither significantly enhanced by the admixture of enzymatically active VCN nor did the DTH response seem to be predictive for a tumor-therapeutic effect . Thomsen-Friedenreich antigens could serologically be detected on untreated cells of Lewis lung adenocarcinoma, B16-melanoma and R-3230 adenocarcinoma . Exposure of Thomsen-Friedenreich antigens after treatment with VCN was enhanced on cells of all tumors except Lewis lung adenocarcinoma . As chessboard vaccination only proved to be successful in Lewis lung adenocarcinoma, but not in the other tumors, it can be concluded that the exposure of Thomsen-Friedenreich antigen plays no decisive role in tumor therapy with tumor cells and VCN . Chessboard vaccination was tolerated without any side effects . Tumor enhancement was not observed.(ABSTRACT TRUNCATED AT 400 WORDS)

Dev Comp Immunol, 1987 Summer, 11(3), 539 - 49
Vibrio anguillarum antigen stimulates mitogenesis and polyclonal activation of salmonid lymphocytes; Yui MA et al.; An antigen preparation of Vibrio anguillarum, a salmonid pathogen, acts as a potent in vitro mitogenic stimulator of splenic and pronephric (anterior kidney) lymphocytes from coho salmon (Oncorhynchus kisutch), chinook salmon (O . tshawytscha) and rainbow trout (Salmo gairdneri) . This antigen (VA) is comparable in its mitogenic activity to Concanavalin A (Con A), Escherichia coli lipopolysaccharide (LPS), and phytohemagglutinin (PHA) . VA gives peak mitogenic responses in coho five days after initiation of cell culture . VA also appears to be a nonspecific polyclonal activator as determined by the generation of plaque forming cells to trinitrophenyl (TNP) and fluorescein (FI) haptenic determinants . Chemical characterization is limited, but it appears that Vibrio LPS could be responsible for these activities.

Microbiol Immunol, 1987, 31(1), 13 - 25
Purification and characterization of Vibrio vulnificus protease; Miyoshi N et al.; A protease was purified from a strain of Vibrio vulnificus isolated from the blood of a septicemic human . The vibrio was cultured in bacto peptone-yeast extract medium, and the protease was purified by a purification procedure including ultrafiltration of the culture supernatant with an Amicon YM 5 membrane, diethylaminoethyl-Sephacel column chromatography, Sephacryl S-200 column chromatography and fast protein liquid chromatography on Mono Q column . The protease preparation revealed homogeneity on polyacrylamide gel electrophoresis and about 30,000-fold purification was achieved, with a yield of about 30% . The isoelectric point of the purified V . vulnificus protease was about 5.80 and its molecular weight was ca . 45,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis . The optimum pH of the protease activity was 8.0 . The V . vulnificus protease was inhibited by a metalloprotease inhibitor and zinc ion and/or ferrous ion were essential for its enzyme activity . No cysteine residue was detected in the V . vulnificus protease . The protease had caseinolytic, elastolytic and collagenolytic activities.

Trans R Soc Trop Med Hyg, 1987, 81(1), 120 - 3
Association between O blood group and occurrence and severity of diarrhoea due to Escherichia coli; Black RE et al.; During studies of diarrhoea due to Escherichia coli in 316 adult volunteers, ABO and Rh blood group determinations were done to look for differences in the occurrence or severity of illness in association with certain blood groups . In studies using heat-labile enterotoxin-producing E . coli, volunteers with O blood group had a significantly higher attack rate for diarrhoea than persons with other blood groups . In contrast, in studies with enteropathogenic or heat-stable enterotoxin-producing E . coli, no association was found between occurrence of diarrhoea and ABO group . These studies, and previous studies finding a similarly increased susceptibility to cholera in persons with O blood group, suggest that the mechanism of increased risk involves an interaction between blood group substances and the similar enterotoxin produced by E . coli and Vibrio cholerae.

Microbiol Immunol, 1987, 31(5), 393 - 401
Inhibitory effect of capsular antigen of Vibrio vulnificus on bactericidal activity of human serum; Shinoda S et al.; Opaque (Op) and translucent (Tr) colonial variants were isolated from Vibrio vulnificus strains . Op-type variants were more resistant than the isogenic Tr-type variants, but the survival rate of the Op-type variants varied with the strains . Antisera were prepared by immunizing rabbit with whole cells of Op and Tr variants of some strains, in which the difference of the sensitivity between Op and Tr cells was remarkable . Then agglutination tests with their living and heat-killed cells were carried out . The results suggested the presence of capsular antigen in Op cells and its absence in Tr cells, with the exception of the existence of a slight amount of capsular material in Tr variants of strain L-180 . The thin capsular layer of Tr cells of strain L-180 was also demonstrated electron microscopically, but the layer was thinner than that of the isogenic Op cells . Results of determination of sugar content in the extracted capsular fraction also showed that Op to Tr transformation was due to loss of capsular antigen of the cells . These results confirmed the morphological studies previously reported which suggested the prevention of host defense system by the capsular material of the vibrio.

Gene, 1987, 53(1), 31 - 40
Extracellular proteins of Vibrio cholerae: molecular cloning, nucleotide sequence and characterization of the deoxyribonuclease (DNase) together with its periplasmic localization in Escherichia coli K-12; Focareta T et al.; The gene encoding the extracellular DNase of Vibrio cholerae was cloned into Escherichia coli K-12 . A maximal coding region of 1.2 kb and a minimal region of 0.6 kb were determined by transposon mutagenesis and deletion analysis . The nucleotide sequence of this region contained a single open reading frame of 690 bp corresponding to a protein of Mr 26,389 with a typical N-terminal signal sequence of 18 aa which, when removed, would give a mature protein of Mr 24,163 . This is in good agreement with the size of 24 kDa, calculated directly by Coomassie blue staining following sodium dodecyl sulphate-polyacrylamide gel electrophoresis and indirectly via a DNA-hydrolysis assay . The protein is located in the periplasmic space of E . coli K-12 unlike in V . cholerae where it is excreted into the extracellular medium . The introduction of the DNase gene into a periplasmic (tolA) leaky mutant of E . coli K-12 facilitates the release of the protein, further confirming the periplasmic location.

Chemotherapy, 1987, 33(6), 428 - 36
Comparison of prophylactic tetracycline and clioquinol in a rabbit model of intestinal infection with Vibrio cholerae and Escherichia coli; Sack DA et al.; The ability of tetracycline and clioquinol to prevent intestinal colonization of Vibrio cholerae and Escherichia coli was tested in a rabbit model . In the model 10(10) bacteria are given via oro-gastric tube following intravenous cimetidine and oral sodium bicarbonate and prior to intraperitoneal tincture of opium . Eighteen hours after challenge the rabbits are sacrificed, and the numbers of the challenge strain remaining in the jejunum and ileum are determined . Tetracycline interrupted the intestinal colonization of V . cholerae and E . coli . Clioquinol however, had minimal effect on the colonization process . Our studies demonstrate the efficacy of prophylactic tetracycline but do not support the use of clioquinol to prevent intestinal infection due to these organisms . This rabbit model may also be useful to study the efficacy of other antibiotics against these bacterial infections.

Bull Soc Pathol Exot Filiales, 1987, 80(5), 761 - 7
{An epidemic of diarrhea caused by Vibrio parahaemolyticus occurring in Abidjan in 1985}; Dosso M et al.; The authors report the epidemiological results of an epidemic about 84 choleriform syndrome in Abidjan (Ivory Coast), June 1985.

Microbiol Immunol, 1987, 31(7), 683 - 9
Stimulation of macrophages by lipopolysaccharide of Vibrio parahaemolyticus; Bandekar JR et al.; The effect of lipopolysaccharide (LPS) from Vibrio parahaemolyticus on the biochemical and phagocytic activities of murine peritoneal macrophages was determined . Intraperitoneal treatment with different doses (0.5-25 micrograms) of V . parahaemolyticus LPS markedly increased the cellular RNA content as well as lysosomal enzyme activities of peritoneal macrophages . The treatment also stimulated the phagocytic activities of macrophages . These observations suggest that V . parahaemolyticus LPS causes stimulation of murine peritoneal macrophages.

Microbios, 1987, 52(211), 87 - 96
A protein preparation from Mycobacterium kansasii culture filtrate has biological activity similar to that of hCG and cholera toxin in human cell lines; Affronti LF et al.; A protein preparation, from culture filtrates from a strain of Mycobacterium kansasii (MK precipitate), which cross reacts with antibodies to human chorionic gonadotropin (hCG) beta subunit and antibodies to cholera toxin beta subunit, has been isolated . A tissue culture assay was used to detect the ability of this preparation to affect the antiviral activity of interferons and to visualize changes in cell shape and cell-cell contact caused by this preparation . A cholera toxin containing precipitate (CT precipitate) from cultures of Vibrio cholerae and commercially prepared hCG were used for comparison . It was found that MK, CT, and hCG may cause reduction or apparent enhancement of interferon antiviral activity or may have no effect . The effect that is observed depends on which test protein the cells are treated with, the type of interferon used, and which cell line is employed in the assay . All three protein preparations had visible effects on the shapes of the cells and the way the cells interacted to form a monolayer.

Gene, 1987, 55(2-3), 197 - 204
A physical map of the chromosomal region determining O-antigen biosynthesis in Vibrio cholerae O1; Ward HM et al.; We have previously described the cosmid cloning of the genes determining the biosynthesis of the Inaba and Ogawa O-antigens of the lipopolysaccharides of Vibrio cholerae O1 (Manning et al., 1986) . By Southern hybridization analysis of chromosomal and cosmid DNA, and heteroduplex analysis between the clones we have been able to precisely define the region of contiguous chromosomal DNA in the vicinity of the O-antigen-encoding region . These data and comparison of end points of clones and of deletion derivatives demonstrate that at least 16 kb of a 19-kb SstI fragment is required to encode O-antigen biosynthesis . Expression of O-antigen is independent of the orientation of this SstI fragment with respect to cloning vectors suggesting that its regulatory region has been cloned intact . No detectable differences were observed in the restriction patterns of the Inaba and Ogawa coding regions implying that only minor changes are involved when serotype conversion (Inaba to Ogawa or vice versa) occurs . Bhaskaran {Ind . J . Med . Res . 47 (1959) 253-260} originally defined this region associated with O-antigen biosynthesis oag; however, to be consistent with other organisms {Hitchcock et al., J . Bacteriol . 166 (1986) 699-705}, it is suggested this be changed to rfb.

Biokhimiia, 1987 Jan, 52(1), 8 - 14
{Similarity of Vibrio alginolyticus, V . cholerae and other Vibrio species with respect to the structure of their flagellar apparatus and ribosomal 5S-RNA}; Bakeeva LE et al.; Electron microscopic analysis of basal bodies of the flagella Vibrio alginolyticus revealed a structure composed of four discs . The diameters of two discs localized in the cytoplasmic membrane appeared to be twice as little as those of the other two discs . In this respect the basal body of V . alginolyticus resembles that of V . cholerae . The 5S sequence of ribosomal RNA from V . alginolyticus appeared to be similar to those of V . cholerae, V . harveyi and some other vibrios . Comparison of 5S-RNA sequence culminated in a dendrogram of evolutionary relationships of various bacterial species, suggesting that V . alginolyticus is a typical representative of the Vibrionacea family . The data obtained are discussed in terms of the role of Na+ energy metabolism in living cells.

Infect Immun, 1987 Jan, 55(1), 269 - 72
Correlation between virulence and colony morphology in Vibrio vulnificus; Simpson LM et al.; Of 38 isolates of Vibrio vulnificus examined, all avirulent strains produced only translucent colonies . All virulent strains, with the exception of biogroup 2 (eel pathogens), exhibited both opaque and translucent colonies . Isogenic morphotypes were examined for a variety of phenotypic and virulence traits . Only the ability to utilize transferrin-bound iron and the presence of a surface polysaccharide were found to correlate with colony opacity and virulence.

Biochem Biophys Res Commun, 1986 Dec 30, 141(3), 1249 - 53
Induction of phage-like particles from a pathogenic strain of Vibrio parahaemolyticus by mitomycin C; Ohnishi T et al.; A rapid drop of absorbance of cell suspensions due to cell lysis was observed midway in cell growth of a pathogenic strain, Vibrio parahaemolyticus WP1, when the cells were incubated in a growth medium containing mitomycin C . We succeeded in isolating phage-like particles from the cell lysate, but not from that of the non-pathogenic strain, WP28 . The purified particle has a clear-cut envelope and is hexagonal in shape with no tail . The size of particles is about 65 millimicrons on the diagonal and about 250 S . The particles contained one kind of protein of 45,000 dalton.

N Engl J Med, 1986 Dec 18, 315(25), 1591 - 5
The acidosis of cholera . Contributions of hyperproteinemia, lactic acidemia, and hyperphosphatemia to an increased serum anion gap; Wang F et al.; To study the metabolic acidosis that occurs during the diarrhea of cholera, we examined the serum anion gap in 21 patients with hypovolemic shock due to Vibrio cholerae infection . Measurements of serum electrolytes, as well as divalent cations and the anionic contributions of serum proteins, lactate, phosphate, and serum creatinine, were made at the time of admission, after rehydration, and during convalescence . At the time of admission, the mean serum concentration of sodium was 134.8 mmol (meq) per liter, that of chloride was 103.2 mmol per liter, and that of bicarbonate was 11.4 mmol per liter; the mean anion gap was 20.2 mmol per liter . The mean serum creatinine concentration was 2.48 mg per deciliter . The low serum bicarbonate level and the high serum anion gap were corrected by rehydration . The increased serum anion gap was caused by hyperproteinemia, lactic acidemia, and hyperphosphatemia, with anionic contributions to the rise in anion gap estimated as protein, 5.5 meq per liter; lactate, 2.5 meq per liter; and phosphate, 2.5 meq per liter . The hyperproteinemia was attributed to dehydration, the lactic acidemia to shock, and the hyperphosphatemia to acidosis and transient renal failure . The mean concentrations of serum calcium and magnesium were slightly elevated but did not affect the increased anion gap . These results indicate that severe cholera causes acidosis with relatively little change in serum chloride but an increased serum anion gap . The acidosis is more profound than would be expected on the basis of stool losses of bicarbonate, because of superimposed lactic acidemia and renal failure.

Appl Environ Microbiol, 1986 Dec, 52(6), 1299 - 304
Rapid serological identification of Vibrio vulnificus by anti-H coagglutination; Simonson J et al.; Staphylococcus aureus Cowan 1 cells were armed with anti-flagellar (anti-H) antibody produced in rabbits immunized with flagellar core protein prepared from Vibrio vulnificus . This reagent was assessed by coagglutination for its capacity to agglutinate and identify V . vulnificus . A species-specific H antigen is expressed in the core proteins of the polar flagella of V . vulnificus . Of 435 V . vulnificus isolates identified bacteriologically, 432 (99.3%) were agglutinated in the slide test within 2 min after the addition of the anti-V . vulnificus H coagglutination reagent . Other than Vibrio pelagius, the reagent did not agglutinate 19 heterologous Vibrio spp . tested, including 290 V . cholerae, 22 V . mimicus, 395 V . parahaemolyticus, and 16 V . fluvialis isolates recovered from seafood and the marine environment . The serological resolution of the coagglutination reaction was enhanced if the organism under test was suspended in 0.1 M Tris buffer-0.1 mM EDTA-1.0% Triton X-100 (TET) for 24 h before serological examination . The TET buffer also increased the sensitivity of the coagglutination reaction 100-fold over that for isolates suspended in 0.3% formalinized phosphate-buffered saline before testing . The anti-H coagglutination test is a rapid, serologically specific, and inexpensive procedure for identifying V . vulnificus one step beyond primary isolation.

J Bacteriol, 1986 Dec, 168(3), 1476 - 8
Spiral conformation of Vibrio cholerae as determined by scanning electron microscopy of elongated cells induced by cephalexin treatment; Konishi H et al.; The elongated cells of Vibrio spp . induced by cephalexin treatment were examined by scanning electron microscopy . The results showed that Vibrio cholerae has a twisted cell body and a right-handed spiral conformation and that the cell bodies of V . parahaemolyticus and V . alginolyticus are straight rather than curved.

Arch Pathol Lab Med, 1986 Dec, 110(12), 1182 - 3
Vibrio cholerae non-01 cellulitis; Gelbart SM et al.; A 46-year-old man presented with pain and fever and a postphlebitic ulcer on his left leg . The wound was suppurative and open at the margins, but there was little underlying fasciitis and no apparent muscle or blood vessel involvement . Three separate wound cultures were obtained at two-day intervals, and all showed only Vibrio cholerae non-01 . The patient was successfully treated with cefazolin sodium . This marks the second documented case of V cholerae non-01 type alone as a causative agent of cellulitis, and the first case where no saltwater origin could be demonstrated.

Southeast Asian J Trop Med Public Health, 1986 Dec, 17(4), 558 - 66
Vibriocidal antibody and antibodies to Vibrio cholerae lipopolysaccharide, cell-bound haemagglutinin and toxin in Thai population; Chongsa-nguan M et al.; Vibriocidal antibody and antibodies to Vibrio cholerae lipopolysaccharide (anti-LPS), cell-bound haemagglutinin (anti-CHA) and toxin (anti-CT) were determined in Thai individuals of various age groups who lived in areas with high (H) and low (L) cholera endemicity . The enzyme-linked immunosorbent assay (ELISA) was performed to detect levels of class specific anti-LPS, anti-CHA and anti-CT . It was found that Thai individuals acquired the vibriocidal antibody early in life . Fifty percent of individuals aged 5 to 15 years old had detectable titre while more than 80% of adults had titres ranged from 1:5 to 1:125 or higher . Thai adults who lived in area with high cholera endemicity had significantly higher vibriocidal antibody levels than their counterparts who lived in area with low cholera endemicity . Lipopolysaccharide was not the only antigen responsible for stimulating the vibriocidal antibody production . Adult levels of all classes of anti-CHA from L were higher than those of H while the anti-LPS in the forms of total immunoglobulins, IgG and IgA were similar but IgM of L was higher than that of H . The levels of all classes of anti-CT from H seemed to increase with age except at the school age (5 years to 15 years old) when there were marked decreases of all antibody classes . Total immunoglobulin and IgM anti-CT at adult age of H and L were not different, although IgG anti-CT of L was higher than that of H and IgA anti-CT of H was higher than that of L.

Mol Gen Genet, 1986 Dec, 205(3), 494 - 500
Nucleotide sequence of ompV, the gene for a major Vibrio cholerae outer membrane protein; Pohlner J et al.; The nucleotide sequence of the ompV gene of Vibrio cholerae was determined . The product of the gene is a 28,000 dalton protein which, after the removal of a 19 amino acid signal sequence, produces a mature outer membrane protein of 26,000 daltons . The cleavage site was determined by amino-terminal amino acid sequencing of the purified mature protein . The DNA upstream of the gene shows the presence of a typical promoter region as judged from the Escherichia coli consensus information; however, the Shine-Dalgarno sequence is associated with a region capable of forming a secondary structure in the mRNA . The formation of this structure would inhibit binding of the mRNA to the ribosome and reduce translation . It is proposed that this structure is recognized by a positive activator in V . cholerae and because of its absence in E . coli ompV is poorly expressed . The distribution of rare codons within ompV suggests that they may serve to slow down the translation of particular domains such that the nascent polypeptide has an opportunity to take up its conformation without interference from the later formed regions . Such a mechanism could aid localization of the protein if export were by a contranslational secretion system.

J Med Microbiol, 1986 Dec, 22(4), 325 - 33
Studies on the Vibrio cholerae mucinase complex . I . Enzymic activities associated with the complex; Stewart-Tull DE et al.; Mucinase enzymes were isolated and partially purified from the culture fluid of Vibrio cholerae grown in proteose peptone-colostrum medium . The mucinase complex contained neuraminidase, endo-beta-N-acetylhexosaminidase, nicotinamide-adenine-dinucleotidase and proteinases . Traces of phospholipase activity were detected but the complex lacked aldolase activity.

Mol Gen Genet, 1986 Dec, 205(3), 501 - 6
Serological properties and processing in Escherichia coli K12 of OmpV fusion proteins of Vibrio cholerae; Pohlner J et al.; Fusion proteins comprising the amino-terminal 99 amino acids of the bacteriophage MS2 replicase and various portions of OmpV a major outer membrane protein of Vibrio cholerae were expressed in Escherichia coli K12 . These fusions were expressed under the control of the PL promoter of bacteriophage lambda, and expression was controlled using a cIts repressor . Fusions occurring within the secretory signal sequence of OmpV gave rise to the production of mature OmpV . The efficiency, however, decreased with progressive deletion of the signal sequence within the fusions . The reactivity of various OmpV fusions with antisera raised against purified OmpV and whole bacteria demonstrated the existence of two antigenic domains: one present in the denatured form and another in the membrane-associated form of OmpV . These domains correspond to markedly hydrophilic regions of the protein as would be predicted for surface-exposed epitopes.

J Neurochem, 1986 Dec, 47(6), 1720 - 7
A ganglioside species (GD1 alpha) migrates at a slow rate and CMP-sialic acid severalfold faster in Xenopus sciatic nerve: fluorographic demonstration; Igarashi M et al.; The ninth dorsal root ganglion of adult Xenopus laevis was labeled with N-acetyl-D-{6-3H}mannosamine, and intraaxonal migration of gangliosides was examined by analysis of the chloroform/methanol extract of each of 5-mm consecutive nerve segments by TLC coupled with fluorography . A unique disialoganglioside (GD1 alpha), which amounted to up to 83% of the total ganglioside in this nerve, migrated at 1-2 mm/day at 15 degrees C . This contrasts with the rapid transport of other ganglioside species previously reported in the optic systems of goldfish, rabbits, chickens, and rats . Fluorographic analysis also revealed a trichloroacetic acid-soluble substance migrating at a velocity of approximately 8 mm/day at 15 degrees C . The substance was considered to be CMP-sialic acid on the basis of observations that it comigrates with authentic CMP-N-acetylneuraminic acid in TLC developed with two different solvent systems, it is very labile to weak acid but resistant to neuraminidase from Vibrio cholerae, it is converted to N-acetylmannosamine when treated first with weak acid and subsequently with N-acetylneuraminic acid aldolase, and it has a beta-sialosyl group in its structure . Because CMP-sialic acid is believed to be the sole sialosyl donor in the cells, its migration in axons toward terminals, together with the previous demonstration of sialyltransferase activity in the synaptosomal plasma membrane, strongly supports the possibility that sialosylation of gangliosides and probably of other sialoglycoproteins is not confined to the Golgi apparatus, but can also occur after the compounds are committed to axonal transport.

J Immunol, 1986 Nov 15, 137(10), 3216 - 23
Binding and mitogenic properties of a galactosyl-specific lectin from the tunicate Didemnum candidum for murine thymocytes and splenocytes; Vasta GR et al.; The affinity-purified major galactosyl-specific lectin (DCL-I) from the tunicate Didemnum candidum binds and induces blastogenesis in untreated and Vibrio cholerae neuraminidase (VCN)-treated murine splenocytes . However, it only binds and stimulates murine thymocytes when they have been pretreated with VCN . Binding and stimulation of VCN-treated splenocytes and thymocytes is specifically inhibited by D-galactose . Binding of lectin to splenocytes is highly increased by the VCN treatment, but the cells are stimulated to the same extent in both untreated and VCN-treated cells . This suggests that although additional DCL-I carbohydrate acceptors are uncovered by VCN treatment of the splenocytes, the ones responsible for triggering blastogenesis are already exposed on the untreated cells . On the contrary, cleavage of terminal sialic acids from the thymocyte cell surface is required to expose DCL-I carbohydrate acceptors in order to obtain a mitogenic response . Our results suggest that the distribution of cell surface galactosyl carbohydrate moieties to which DCL-I binds is different in thymocytes and splenocytes.

Can J Microbiol, 1986 Nov, 32(11), 889 - 91
Role of chitin in the survival of Vibrio parahaemolyticus at different temperatures; Karunasagar I et al.; Washed cells of Vibrio parahaemolyticus declined in numbers when incubated in phosphate-buffered saline for 6 h at different temperatures . Addition of chitin flakes to phosphate-buffered saline not only helped the organism to survive, but also resulted in an increase in cell numbers, particularly at 10 degrees C . The effect of chitin could not be simulated by N-acetylglucosamine, yeast extract, starch, or casein.

J Appl Bacteriol, 1986 Nov, 61(5), 469 - 80
A probability matrix for the identification of species of Vibrio and related genera; Bryant TN et al.; A matrix for the probabilistic identification of species of Vibrio and related genera has been constructed using the data from 1091 strains collected throughout the world and classified . Thirty-eight phenons are included in the matrix, 31 of these represent previously identified species or biovars and seven represent phenons which could not be identified and may represent new species . The identification matrix incorporates 81 characters although a subset of 30 tests can be used to distinguish the 38 phenons from each other . The additional 51 tests were included to assist the identification of some strains for which the initial 30 tests were inadequate . No significant cluster overlap was found at the 5% level and the identification score for the Hypothetical Median Organism of each cluster exceeded 0.9999 in all cases.

J Appl Bacteriol, 1986 Nov, 61(5), 437 - 67
Numerical classification of species of Vibrio and related genera; Bryant TN et al.; Data from 1091 strains of the family Vibrionaceae collected in five different studies have been merged into a single data matrix and analysed in a taxonomic study . A set of 142 characters was selected to compare these data . Seventy-nine characters were common to all studies, but data for the other 63 characters were incomplete . Cultures of 90 strains, examined in more than one of the original studies, were used to estimate test error and inter-study variability . The data from these replicate strains also allowed the problem of merging data from different studies to be assessed . Taxonomic resemblance was estimated on the basis of 111 characters using the SSM coefficient and UPGMA clustering . A taxonomic analysis based on 999 strains, which included most of the major species of the family Vibrionaceae, gave 59 clusters and 44 unclustered strains . A table of properties of these phenons was produced . The results showed that data obtained from studies carried out at different times and in different locations, but using standard techniques, could be combined and used to provide useful taxonomic information.

Appl Environ Microbiol, 1986 Nov, 52(5), 1218 - 20
Demonstration of a plasmid-borne gene encoding a thermostable direct hemolysin in Vibrio cholerae non-O1 strains; Honda T et al.; Of 15 Vibrio cholerae non-O1 strains, 2 produced a hemolysin termed NAG-rTDH, which is very similar to the thermostable direct hemolysin of V . parahaemolyticus . These two strains contained DNA sequences which are homologous to a DNA probe for the V . parahaemolyticus thermostable direct hemolysin gene . A probe-positive 9-kilobase HindIII fragment was cloned from a plasmid of a V . cholerae non-O1 strain into plasmid pBR322, and the resulting Escherichia coli clones produced intracellular NAG-rTDH.

Appl Environ Microbiol, 1986 Nov, 52(5), 1209 - 11
Bioluminescence in a strain of the human pathogenic bacterium Vibrio vulnificus; Oliver JD et al.; We report the existence of a bioluminescent strain of the human pathogen Vibrio vulnificus . The isolate was obtained from blood following a fatal wound infection and thus represents the first description of an infection caused by a luminescent bacterium.

J Clin Microbiol, 1986 Nov, 24(5), 744 - 8
GM1 erythroimmunoassay for detection and titration of Escherichia coli heat-labile enterotoxin; Germani Y et al.; A GM1 ganglioside erythroimmunoassay for the detection of heat-labile Escherichia coli enterotoxin (LT) was developed for use in poorly equipped laboratories in developing countries . This assay is based on the immunological similarity between Vibrio cholerae toxin and LT and uses cholera toxin antiserum and sheep anti-rabbit immunoglobulin covalently coupled to sheep erythrocytes as conjugate . This assay has the following advantages over other currently available techniques: the reagents it uses are stable, in particular, tanned and sensitized sheep erythrocytes; GM1 ganglioside is commercially available; erythro-adsorption can be read with the naked eye; the test can be completed in 1 day; and as little as 4 ng of V . cholerae toxin or LT per ml can be detected accurately . The GM1 ganglioside erythroimmunoassay showed good quantitative and qualitative correlation with the Vero cell assay and the conventional GM1 enzyme-linked immunosorbent assay . The GM1 ganglioside erythroimmunoassay was somewhat less sensitive than the GM1 enzyme-linked immunosorbent assay but more sensitive than the Vero cell assay . Results obtained for 12 LT-positive and 138 LT-negative E . coli strains correlated with results obtained with GM1 enzyme-linked immunosorbent and Vero cell assays.

Infect Immun, 1986 Nov, 54(2), 415 - 20
Hemolysin production and cloning of two hemolysin determinants from classical Vibrio cholerae; Richardson K et al.; The hemolytic activity of 20 classical and 3 El Tor strains of V . cholerae O1 was examined phenotypically and genetically . The El Tor strains lysed bovine, chicken, human, rabbit, and sheep erythrocytes (RBCs), while the classical strains lysed only chicken and rabbit RBCs . The assay was done with RBCs in Tris-NaCl buffer, since phosphate-buffered saline was found to inhibit hemolytic activity . Hemolytic activity in culture supernatants from El Tor strains was more sensitive to heat inactivation than that in supernatants from the classical strain 395 . A gene library of strain 395 was examined for hemolytic activity, and two distinct hemolytic clones were identified . One clone appeared identical to the previously cloned hemolysin structural gene from El Tor V . cholerae, while the other did not hybridize to the El Tor hemolysin probe, had a unique restriction enzyme digestion pattern, and encoded a hemolysin whose activity differed from that of the El Tor hemolysin clones . We suggest that the hemolysin specified by the determinant originally cloned from an El Tor vibrio be designated hemolysin I and the second hemolysin, cloned from the classical vibrio, be designated hemolysin II.

J Hosp Infect, 1986 Nov, 8(3), 275 - 82
An outbreak of nosocomial cholera in a rural Bangladesh hospital; Ryder RW et al.; At a time of year when Vibrio cholerae infection accounted for over 50% of admissions to a rural Bangladeshi diarrhoea treatment centre, 29% of 48 patients hospitalized with non-cholera diarrhoea developed nosocomial V . cholerae infection . During an investigation of the 8-week outbreak, only the severity of the non-cholera diarrhoea which prompted hospital admission emerged as an important risk factor for nosocomial infection; food, intravenous solutions, oral rehydration fluid, patient attendants and hospital personnel could not be implicated as transmission sources . Patients receiving antibiotics while hospitalized did not develop nosocomial infection . Antecedent non-cholera diarrhoea may represent an important risk factor in some cases of V . cholerae infection occurring in persons who reside in cholera-endemic areas where rates of non-cholera diarrhoea are also high.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Oct, (10), 59 - 62
{Experimental research on cholera vaccines . Local immunity}; Manakhilov RB et al.; Parenteral immunization of rabbits with cholera vaccine decreased the number of Vibrio cholerae adhering to the mucous membrane of the small intestine . Cholera toxoid and the complex preparation ensure protection from the local action of cholera toxin on the ligated loop of the rabbit intestine, while cholera vaccine produces no effect under the same conditions . The use of three preparations under study leads to the decrease of exudative reaction to the introduction of live V . cholerae, the effectiveness of these vaccines growing in the following order: cholera vaccine, cholera toxoid, the complex preparation.

J Trop Med Hyg, 1986 Oct, 89(5), 269 - 76
The characterization of Vibrio cholerae isolated in Kenya in 1983; Ichinose Y et al.; A total of 245 strains of Vibrio cholerae 01 and two strains of V . cholerae non-01 were isolated and collected from diarrhoeal patients in Homa Bay District Hospital and the other medical facilities in Nyanza Province, Kenya in 1983 . The majority of V . cholerae 01 tested were Ogawa type (with the exception of nine Inaba type), biotype E1 Tor (except one untypable strain) and Celebes original type (except one cured type) . Haemolytic activity to sheep red blood cells was detected in 75.5% of isolates . Out of 245 strains of V . cholerae 01, 184 were resistant to tetracycline, streptomycin and ampicillin . All were sensitive to chloramphenicol and nalidixic acid . Only one strain of V . cholerae 01 was sensitive to all five antimicrobial agents tested . An environmental cholera survey was done after the cholera outbreak subsided . Twenty strains of V . cholerae non-01 were isolated from water samples in Nyanza Province but none of V . cholerae 01 was isolated.

Antimicrob Agents Chemother, 1986 Oct, 30(4), 622 - 3
Antibiotic resistance patterns of Vibrio mimicus isolated from human and environmental sources in Bangladesh; Chowdhury MA et al.; Twenty-five environmental and 19 clinical strains of Vibrio mimicus were tested for antibiotic susceptibility patterns . Environmental strains were resistant to streptomycin, kanamycin, and trimethoprim-sulfamethoxazole; clinical strains were susceptible . Environmental strains showed variable resistance to ampicillin (44%), but clinical strains were susceptible . All strains tested were susceptible to chloramphenicol and gentamicin.

Appl Environ Microbiol, 1986 Oct, 52(4), 875 - 9
Comparative study of expression of hemagglutinins, hemolysins, and enterotoxins by clinical and environmental isolates of non-O1 Vibrio cholerae in relation to their enteropathogenicity; Datta-Roy K et al.; A comparative study was undertaken of clinical and environmental isolates of non-O1 Vibrio cholerae with respect to their hemagglutinating, hemolytic, enterotoxigenic, and enteropathogenic activities . Cell-associated hemagglutinin titers of the clinical and environmental isolates did not differ much, although the clinical isolates displayed higher cell-free hemagglutinin titers compared with those of environmental isolates . Culture supernatants of 61.5% (24 of 39) of clinical isolates showed hemolytic activity (greater than or equal to 10% lysis of rabbit erythrocytes), while only 33.3% (10 to 30) of the environmental group had such activity . Furthermore, hemolytic activities of the clinical isolates showed a good correlation with their cell-associated hemagglutinin titers which was not true for the environmental group . Culture supernatants of 45.8% (11 of 25) of the clinical and 20% (2 of 10) of the environmental isolates exhibited enterotoxigenic activity in the rabbit ileal loop assay . Such activity was mediated mainly by cholera toxin-like substances, although some of the isolates produced fluid-accumulating factors unrelated to cholera toxin . Experimental animal studies demonstrated that the enteropathogenic potential of the environmental isolates was significantly lower than that of the clinical group . Further analysis of our data showed that phenotypic expression of cholera toxin-like products by the non-O1 V . cholerae isolates was accompanied by their enteropathogenicity . The latter effect was also noted with some of the cholera toxin-negative isolates, particularly in those having high hemagglutinating and hemolytic titers.

Appl Environ Microbiol, 1986 Oct, 52(4), 824 - 31
Behavior of Vibrio cholerae in hot foods; Makukutu CA et al.; Four food types held hot at 45 to 60 degrees C were deliberately contaminated with O1 and non-O1 Vibrio cholerae strains . These organisms were assayed for survival and recovery from the foods within 1 h of the time the food was kept hot . The results showed no growth of V . cholerae non-O1 on thiosulfate-citrate bile-sucrose agar plates after 24 h of incubation at 37 degrees C for food held hot at 50 to 60 degrees C . Growth was low for V . cholerae O1 and was not achieved in some instances in which foods were held at either 55 or 60 degrees C after 40 or 60 min of from the time the food was kept hot . Both organisms, however, were recovered equally from all food types held at all temperatures after 48 h of incubation . When incubated for an additional 24 h, the organisms grew to unusually small-sized colonies, measuring 0.1 to 0.3 mm in diameter, on the same agar plates that were negative for growth after an initial 24 h of incubation . It was concluded that V . cholerae survived the period of time held at hot temperatures . Although the organisms were not recovered from some foods when held at some of the temperatures and times after 24 h of incubation, they remained viable . An incubation period of 48 h at 37 degrees C was found to be appropriate for the recovery of V . cholerae from hot foods.

Appl Environ Microbiol, 1986 Oct, 52(4), 794 - 801
Phospholipid ester-linked fatty acid profile changes during nutrient deprivation of Vibrio cholerae: increases in the trans/cis ratio and proportions of cyclopropyl fatty acids; Guckert JB et al.; The phospholipid ester-linked fatty acids of 0-day-, 7-day-, and 30-day-starved cultures of Vibrio cholerae were compared . Statistically significant trends were noted in the fatty acid profiles as the cells starved . The amount of the cis-monoenoic fatty acids declined (e.g., 16:1 omega 7c: 0 day, 39%; 7 day, 18%; 30 day, 11%) . In contrast, the saturated fatty acids, the cyclopropyl derivatives of the cis-monoenoic fatty acids, and trans-monoenoic fatty acids increased during starvation . For instance, the amounts of 16:1 omega 7t were: 0 day, 1%; 7 day, 13%; 30 day, 17%; which increased the trans/cis ratio for 16:1 omega 7 from 0.02 (0 day) to 0.70 (7 day) to 1.56 (30 day) . This may be due to the reported high turnover rates of cis-monoenoic fatty acids of membrane phospholipids and the availability of enzymes for the metabolism of these isomers . During starvation-induced phospholipid loss, the cis-monoenoic fatty acids would, therefore, be preferentially utilized . The ability to either synthesize trans-monoenoic acids (which are not easily metabolized by bacteria) or modify the more volatile cis-monoenoic acids to their cyclopropyl derivatives may be a survival mechanism which helps maintain a functional (although structurally altered) membrane during starvation-induced lipid utilization . In addition, a trans/cis fatty acid ratio significantly greater than that reported for most bacterial cultures and environmental samples (less than 0.1) may be used as a starvation or stress lipid index . Such a ratio could help determine the nutritional status of ultramicrobacteria and other reported dormant cells in natural aquatic environments.(ABSTRACT TRUNCATED AT 250 WORDS)

Zh Mikrobiol Epidemiol Immunobiol, 1986 Oct, (10), 96 - 9
{Effect of intestinal microflora on the pathogenesis of cholera intoxication}; Nazarova LS et al.; Germ-free minipigs, previously treated with bacteroids, develop cholinergic reaction after the intragastric administration of Vibrio cholerae exotoxin . The intensity of this reaction, disturbances in homeostasis, and the character of morphological changes depend on the dose of choleragen, the bacteroid strain, and the presence of the concomitant (Escherichia coli) and residual microflora in the intestine.

Microb Pathog, 1986 Oct, 1(5), 425 - 32
Comparison of the nucleotide sequences of the genes for the thermostable direct hemolysin and the thermolabile hemolysin from Vibrio parahaemolyticus; Taniguchi H et al.; The nucleotide sequences of genes encoding the thermostable direct (TSD) hemolysin and the thermolabile (TL) hemolysin of Vibrio parahaemolyticus were determined . From the nucleotide sequence of the TSD hemolysin gene, it was revealed that the preprotein and the mature protein consisted of 189 amino acids and 165 amino acids, and that the molecular weights were 21.1 kDa or 18.5 kDa, respectively . Our data regarding TSD hemolysin were in complete agreement with previously published data . From the nucleotide sequence of the TL hemolysin gene, it was revealed that the preprotein and the mature protein consisted of 418 amino acids and 398 amino acids, and that the molecular weights were 47.5 kDa and 45.3 kDa, respectively . The GC content of the TSD hemolysin gene was 35.6%, while that of the TL hemolysin gene was 47.6% which is almost the same as that of V . parahaemolyticus genome . Maxicell analysis revealed that the molecular weights of the proteins encoded by the TSD hemolysin gene were 22.0 and 19.5 kDa, and that of the protein encoded by the TL hemolysin gene was 45.5 kDa, and that the promoters of these two hemolysin genes of V . parahaemolyticus were functional in Escherichia coli.

Microbiologia, 1986 Oct, 2(2), 121 - 8
{DNA-DNA homology and phenotypic characterization of environmental strains of Vibrio parahaemolyticus and Vibrio pelagius}; Pujalte MJ et al.; A taxonomic study by means of DNA homology and the most recent phenetic criteria has been carried out in 11 environmental strains of Vibrio parahaemolyticus and 4 of Vibrio pelagius, previously ascribed to the respective phenons in a numerical taxonomy study . The strains of V . parahaemolyticus showed genetic homogeneity, a guanine plus cytosine content (G + C) of 46% and 61% homology with V . parahaemolyticus type strain ATTC 17802 . They exhibited phenetic atypicalities but could be ascribed to the species V . parahaemolyticus . The strains of V . pelagius behaved similarly in regard to the phenotype, but were genetically heterogeneous; they showed a % G + C of 44-46 and were assigned to V . splendidus-V . pelagius homology group.

J Gen Microbiol, 1986 Oct, 132 ( Pt 10), 2949 - 52
Evidence for the role of a siderophore in promoting Vibrio anguillarum infections; Wolf MK et al.; Vibrio anguillarum strain 775 harbouring the virulence plasmid pJM1 produces a plasmid-mediated siderophore that can crossfeed siderophore-deficient, receptor-proficient mutants of V . anguillarum in vitro . Experimental infections of salmonid fishes with mixtures consisting of the wild-type strain and a siderophore-deficient, receptor-proficient mutant resulted in recovery of both the wild-type and the mutant strain, while in infections with mixtures consisting of the wild-type strain and a siderophore-deficient, receptor-deficient mutant only the wild-type strain could be recovered . These results suggest that the V . anguillarum plasmid-mediated siderophore is produced in vivo in a diffusible form and that it is an important factor of virulence.

Arch Microbiol, 1986 Oct, 146(1), 30 - 4
Temperature and oxygen regulated expression of a glutamine synthetase gene from Vibrio alginolyticus cloned in Escherichia coli; Maharaj R et al.; Glutamine synthetase (GS) synthesis in Vibrio alginolyticus was regulated by temperature, oxygen and nitrogen levels . A GS gene, glnA from V . alginolyticus was cloned on a 5.67 kb insert in the recombinant plasmid pRM210, which enabled Escherichia coli glnA, ntrB, ntrC deletion mutants to utilize (NH4)2SO4 as a sole source of nitrogen . The V . alginolyticus glnA gene was expressed from a regulatory region contained within the cloned fragment . V . alginolyticus glnA expression from pRM210 was subject to regulation by temperature, oxygen and nitrogen levels . GS specific activity in an E . coli wild-type strain was not affected by temperature or oxygen . pRM211 was a deletion derivative of pRM210 and GS production by pRM211 was not regulated by temperature, oxygen or nitrogen levels in E . coli.

Appl Environ Microbiol, 1986 Oct, 52(4), 788 - 93
Effect of nutrient deprivation on lipid, carbohydrate, DNA, RNA, and protein levels in Vibrio cholerae; Hood MA et al.; The response of Vibrio cholerae to low nutrient levels was determined by measuring the concentrations of lipids, carbohydrates, DNA, RNA, and proteins over a 30-day starvation period . Ultrastructural integrity was observed by transmission electron microscopy . Total lipids and carbohydrates declined rapidly within the first 7 days, while DNA and protein exhibited a more constant decline over the 30 days of starvation . In contrast, RNA showed little decrease upon starvation . Although neutral lipids were lost, the percentage of neutral lipids did not decline as rapidly as the phospholipids . Detectable levels of poly-beta-hydroxybutyrate disappeared completely by 7 days . Carbohydrate profiles revealed the relative loss of the five-carbon sugar ribose and N-acetylglucosamine and a relative increase in the total six-carbon sugars, especially glucose . Morphologically, ribosomes appeared to exhibit no structural change, while inclusion bodies and mesosomelike structures disappeared completely, and cell wall and membrane integrity was lost . The data suggest that V . cholerae differs somewhat from other marine vibrios in its response to low nutrients but shares some characteristics in common with them . The data also suggest that certain lipids and carbohydrates may provide the endogenous energy sources needed for dormancy preparation and cell maintenance under nutrient starvation.

J Gen Microbiol, 1986 Sep, 132 ( Pt 9), 2601 - 3
Metabolic reactions responsible for glucose stimulation of alkaline phosphatase in Vibrio cholerae; Mitra S et al.; Alkaline phosphatase activity in Vibrio cholerae strain 569B grown in low-phosphate medium was stimulated if glucose or glycerol was used as the carbon source . No such stimulation was observed, however, if tricarboxylic acid cycle intermediates like succinate or citrate were used . Experiments using specific enzyme inhibitors strongly indicated that the metabolic reactions of the glycolytic pathway from glyceraldehyde 3-phosphate to 2-phosphoglycerate play a key role in the stimulation process.

Appl Environ Microbiol, 1986 Sep, 52(3), 586 - 8
Colonization of the gut of the blue crab (Callinectes sapidus) by Vibrio cholerae; Huq A et al.; Attachment of Vibrio cholerae to the mucosal surface of the intestine is considered to be an important virulence characteristic . Vibrio cholerae, an autochthonous member of brackish water and estuarine bacterial communities, also attaches to crustacea, a significant factor in multiplication and survival of V . cholerae in nature . The ability of V . cholerae to attach to the gut wall of the blue crab (Callinectes sapidus) was examined, and attachment was observed only in the hindgut and not the midgut of crabs, confirming a requirement for chitin in the attachment of V . cholerae to invertebrate and zooplankton surfaces . The new finding of attachment of V . cholerae to the hindgut of crabs may be correlated with the epidemiology and transmission of cholera in the aquatic environment . The crab model may also prove useful in elucidating the mechanism(s) of ion transport in crustacea.

Appl Environ Microbiol, 1986 Sep, 52(3), 583 - 5
Evaluation of methods for enumeration of Vibrio parahaemolyticus from seafood; Karunasagar I et al.; The efficiency of several enrichment broths in recovering Vibrio parahaemolyticus inoculated into fish homogenates was studied . Recovery by the most probable number technique was very low in all the broths, while direct plating on thiosulfate citrate bile salt sucrose agar yielded better recovery . A decrease in the enrichment time to 8 from 18 h did not improve recovery . At concentrations exceeding 2.5 micrograms/ml, polymyxin was inhibitory to V . parahaemolyticus.

Biull Eksp Biol Med, 1986 Sep, 102(9), 342 - 5
{Resistance of the alimentary canal of Daphnia magna Straus to the effect of enteropathogenic NAG-vibrios}; Avtsyn AP et al.; The interrelations between 30 Daphnias and enteropathogenic NAG-vibrios were experimentally investigated . Using histological and immunoserological techniques, NAG-vibrio administration into the medium inhabited by Crustaceous was shown to cause no pathological changes in the animal alimentary canal . Daphnias used these organisms as food . The intestinal epithelium is well protected from mechanical injury by peritrophic membrane, chitinous lining and peritrophic cavity where digestion takes place.

J Med Microbiol, 1986 Sep, 22(2), 133 - 41
Antibacterial immunity to Vibrio cholerae in rats; Cooper GN et al.; Blind loops prepared in the small intestines of fasted, MgSO4-treated rats were shown to provide a simple, consistent and inexpensive means of studying mucosal colonisation by Vibrio cholerae serogroup O1 . When c . 2000 cfu were injected, the number of mucosa-associated V . cholerae in each loop increased by c . 5-6 orders of magnitude in 10-14 h, without enterotoxin-induced fluid production . Scanning electronmicroscopy and culture suggested that most surface-associated organisms were present in the adherent surface mucus . V . cholerae strains varied in terms of surface-colonising capacity . Immunisation with V . cholerae given intra-intestinally greatly reduced the rate of increase and final number of mucosa-associated vibrios within the 14-h period after challenge . The method could be used to compare the immunity induced by various immunising regimens . Immunity was sometimes accompanied by intestinal mucus-borne antibody against V . cholerae lipopolysaccharide but was sometimes demonstrated in the absence of such antibody or of mucus-borne antibody to heat-sensitive surface protein.

Infect Immun, 1986 Sep, 53(3), 700 - 1
Inhibition of cholera toxin production by thiols in Vibrio cholerae; Shimamura T et al.; We found that thiols reduced the amount of cholera toxin produced by Vibrio cholerae 569B in vitro . A sulfhydryl group at least was necessary for the reduction of cholera toxin production by thiols.

Diagn Microbiol Infect Dis, 1986 Sep, 5(3), 221 - 3
Report of a wound infection caused by Vibrio parahaemolyticus and Vibrio vulnificus; McMeeking AA et al.; The present case describes a foot wound caused by a clam shell from which both Vibrio parahaemolyticus and Vibrio vulnificus were recovered . Although extraintestinal infections associated with Vibrio parahaemolyticus have been reported previously, the simultaneous isolation of two marine vibrios from our case suggests that these organisms may coexist in mixed infections from a common source.

FEBS Lett, 1986 Aug 18, 204(2), 336 - 40
Protein-chemical analysis of pertussis toxin reveals homology between the subunits S2 and S3, between S1 and the A chains of enterotoxins of Vibrio cholerae and Escherichia coli and identifies S2 as the haptoglobin-binding subunit; Capiau C et al.; The purified toxin of Bordetella pertussis was dissociated in 5 M urea in the presence of immobilized haptoglobin . The toxin was dissociated in free S1, free S5 and the free complexes S2-S4 and S3-S4, with S2-S4 as the only haptoglobin-binding moiety, identifying S2 as the haptoglobin-binding protein . Partial NH2-terminal amino acid sequences were obtained from the dissimilar toxin subunits, after separation by SDS-polyacrylamide gel electrophoresis followed by electroblotting onto polybrene-coated glass-fiber sheets . The sequences reveal extensive homology of the N-terminal portions of the constitutive subunits S2 and S3 and between S1 and the enterotoxin A chains of Vibrio cholerae and Escherichia coli.

J Gen Microbiol, 1986 Aug, 132 ( Pt 8), 2235 - 42
Composition and immunochemical properties of the cell surface proteins of Vibrio cholerae; Kabir S; The composition and immunochemical properties of cell surface proteins of Vibrio cholerae belonging to both the biotypes (classical and El Tor) and the serotypes (Ogawa and Inaba) were investigated . Proteins were isolated by extraction with EDTA/NaCl . When the extract was further treated with sodium deoxycholate, a product significantly enriched with the major protein was obtained . The surface localization of these proteins was confirmed by immunoelectron microscopy using protein A-colloidal gold particles as probes . Antisera to these proteins possessed complement-mediated bactericidal activities towards V . cholerae strains belonging to both the biotypes and the serotypes, and upon crossed immunoelectrophoresis produced several immunoprecipitation reactions towards whole-cell sonicates belonging to all types of V . cholerae . These proteins were immunogenic in the rabbit intestine, as antibodies of two classes (IgG and IgA) were detected in the intestinal fluids . The intestinal immune response was greatly enhanced when cell surface proteins were administered with liposomes . These results suggest that cell surface proteins represent common antigens of V . cholerae and can be explored as vaccine candidates against cholera.

J Gen Microbiol, 1986 Aug, 132 ( Pt 8), 2225 - 34
Molecular cloning of a gene coding for a Vibrio cholerae haemagglutinin; Van Dongen WM et al.; Recombinant plasmids encoding a Vibrio cholerae haemagglutinin were isolated from the highly virulent V . cholerae strain C5 by cosmid cloning . Both Escherichia coli HB101 containing the recombinant plasmids and V . cholerae C5 were able to agglutinate a variety of erythrocytes from human and animal origin; this haemagglutination was not inhibited by D-mannose or L-fucose . Subcloning of the recombinant cosmid DNA revealed that a 1.3 kb DNA fragment was sufficient for haemagglutinin production in E . coli HB101 . Under direction of this 1.3 kb Vibrio DNA fragment, two proteins were made in E . coli minicells, of 27 and 10 kDa . Haemagglutinin-encoding sequences were not detected in every V . cholerae strain.

Appl Environ Microbiol, 1986 Aug, 52(2), 297 - 301
Effects of microcosm salinity and organic substrate concentration on production of Vibrio cholerae enterotoxin; Tamplin ML et al.; The effects of aquatic processes on production of cholera toxin by Vibrio cholerae were studied with seawater microcosms . Several salinity and organic nutrient concentrations were employed . At 10 g of organic nutrient per liter of seawater, toxin production increased as the salinity was increased . At lower organic nutrient concentrations, toxin production was markedly enhanced when the salinity was 20 and 25% . Toxin concentration increased with salinity, independent of cell concentration and toxin stability . From the results obtained in this study, it is concluded that physical and chemical parameters of the aquatic environment affect not only the physiological state of V . cholerae, but also its potential pathogenicity.

Clin Exp Immunol, 1986 Aug, 65(2), 443 - 9
Assay dependent specificities of monoclonal antibodies to bacterial antigens; Ghosh S et al.; Six rat monoclonal antibodies, all of the IgG2b class, were generated from rats immunized with the 35A3 (Inaba) and NIH-41 (Ogawa) strain of Vibrio cholerae and selected by enzyme-linked immunosorbent assay (ELISA) on the whole organisms . When the fine specificity was dissected by several different immunological assays, the antibodies could be divided into three groups, each with a different specificity profile . Two antibodies were totally specific to the Ogawa serotype on all assays, three had a preference for Inaba but could be shown to display assay dependent cross reactions of variable intensity with Ogawa . The sixth showed total specificity for Ogawa on some assay systems, apparent total specificity for Inaba on others, and variable reaction with both serotypes on yet other assay systems . The data emphasize that it is possible to produce antibodies which do not conform to the conventional serological classification of antigens and that specificity is highly dependent on method of assessment.

J Clin Microbiol, 1986 Aug, 24(2), 203 - 9
Saliva, breast milk, and serum antibody responses as indirect measures of intestinal immunity after oral cholera vaccination or natural disease; Jertborn M et al.; The possibility that antibody responses in serum, saliva, or breast milk samples to oral vaccines or enteric infections may reflect the intestinal immune response was evaluated in Bangladeshi volunteers orally immunized with a cholera B subunit-whole-cell vaccine (B + WCV) and in patients convalescing from enterotoxin-induced diarrheal disease . Two peroral doses of B + WCV induced antitoxin and antibacterial antibody responses in the intestinal fluids of 76 and 92%, respectively, of the volunteers and in serum samples in 90 and 69% of those tested . These responses were comparable to those obtained after cholera or enterotoxigenic Escherichia coli disease . Whereas immunoglobulin A (IgA) antitoxin titer increases in saliva (44%) and breast milk (29%) specimens after vaccination were less frequent than in intestinal fluid (76%), antitoxin responses in saliva and breast milk occurred in 80 to 90% of the patients after disease . Also, antilipopolysaccharide (anti-LPS) titer increases in extraintestinal body fluids were found more frequently after disease than after vaccination . A comparison of the frequency and magnitude of antibody response in different body fluids with those in intestinal lavage fluid revealed no extraintestinal antibody that directly reflected the intestinal immunity . However, comparison of vibriocidal and IgG antitoxin antibodies in serum specimens with antitoxin and anti-LPS IgA responses in intestinal fluids after the vaccination of volunteers showed a sensitivity of 70 to 90% and a predictive accuracy of about 80% for the serum analyses reflecting the intestinal immune responses . Furthermore, antitoxin and anti-LPS antibody responses in saliva and breast milk samples seemed to be useful proxy indicators of a gut mucosal response of these antibodies after enterotoxin-induced diarrheal disease showing sensitivity vales of 70 to 90% and predictive accuracy vales of 70 to 100%.

Microb Pathog, 1986 Aug, 1(4), 361 - 71
Strong biotype and serotype cross-protective antibacterial and antitoxic immunity in rabbits after cholera infection; Lycke N et al.; We studied whether immunity evoked by infection with classical or El Tor V . cholerae 01 organisms in rabbits (the RITARD model) also gives protection against cholera caused by V . cholerae of heterologous biotype as well as serotype, and whether such protection is antibacterial and/or antitoxic . A primary infection with a classical Ogawa or El Tor Inaba strain resulted in intestinal colonization and diarrheal disease in a dose-related manner though the El Tor strain was more virulent . As few as 10(3) El Tor organisms gave disease in more than 90% of the rabbits as compared to 10(9) classical organisms (ED90); the El Tor strain also gave rise to diarrhea with earlier onset and of greater severity and longer duration . The primary infection induced strong protective immunity against later challenge with either the homologous or the heterologous strain in doses that corresponded to 1,000 x ED90 . Protection was associated with marked inhibition of colonization, and when rabbits convalescing from cholera infection were challenged with graded doses of bacteria or purified toxin in ligated intestinal loops significant antibacterial as well as antitoxic immunity was evident . Titer rises in serum vibriocidal and anti-lipopolysaccharide antibodies were similar after infection with either strain, whilst antitoxin titer rises were more marked after El Tor infection . During infection V . cholerae 01 organisms seem to express protective antigens that stimulate immunity which extends across both biotype and serotype barriers.

Antimicrob Agents Chemother, 1986 Aug, 30(2), 245 - 7
Plasmid penicillin resistance in Vibrio cholerae: identification of new beta-lactamase SAR-1; Reid AJ et al.; Two strains of Vibrio cholerae biotype El Tor, isolated in Tanzania, possessed a single IncC resistance plasmid of 113 kilobases . Both plasmids encoded the production of a novel beta-lactamase, SAR-1, which was 33,700 daltons in size and was able to hydrolyze carbenicillin as well as penicillin G . The SAR-1 beta-lactamase was quite distinct from all other plasmid beta-lactamases by virtue of its unusually low isoelectric point and a combination of its size, substrate profile, and inhibition properties . This enzyme is only the second beta-lactamase identified in V . cholerae species and the first to be reported in V . cholerae strains isolated in Southern Africa.

Can J Microbiol, 1986 Aug, 32(8), 632 - 6
Purification and partial characterization of a Vibrio hollisae hemolysin that relates to the thermostable direct hemolysin of Vibrio parahaemolyticus; Yoh M et al.; Hemolysin produced by Vibrio hollisae (Vh-rTDH), which is related to the thermostable direct hemolysin (Vp-TDH) of Vibrio parahaemolyticus, was studied . Vh-rTDH was purified by successive column chromatographies on diethylaminoethyl-cellulose and an immunoaffinity column coupled with anti Vp-TDH immunoglobulin . The purified toxin was homogeneous, as demonstrated by conventional and sodium dodecyl sulfate--polyacrylamide gel electrophoresis (PAGE) . The molecular weight of Vh-rTDH was slightly smaller than that of Vp-TDH, as determined by sodium dodecyl sulfate--slab gel electrophoresis . Conventional PAGE also showed a difference between Vh-rTDH and Vp-TDH . Vp-TDH and Vh-rTDH showed different lytic activities on erythrocytes from various animals, in particular chicken, sheep, and calf . The hemolytic activity of Vh-rTDH was heat labile when heated at 70 degrees C for 10 min, unlike Vp-TDH . Immunological cross-reactivity between Vh-rTDH and Vp-TDH was demonstrated by both the Ouchterlony test and neutralization test.

Carbohydr Res, 1986 Aug 1, 150, 213 - 9
Structural studies of the O-antigen from Vibrio cholerae O:21; Ansari AA et al.; The O-antigen from Vibrio cholerae O:21 has been investigated, using n.m.r . spectroscopy, methylation analysis, and Smith degradation as the main methods . It is concluded that the O-antigen is composed of tetrasaccharide repeating-units having the following structure (in which Hep = D-glycero-D-manno-heptose) . (Formula: see text).

Infect Immun, 1986 Aug, 53(2), 272 - 7
Molecular cloning and expression in Escherichia coli K-12 of the O antigens of the Inaba and Ogawa serotypes of the Vibrio cholerae O1 lipopolysaccharides and their potential for vaccine development; Manning PA et al.; The gene clusters that determine the biosynthesis of both the Inaba and Ogawa serotypes of the O antigen of the lipopolysaccharide of Vibrio cholerae were cloned and expressed in Escherichia coli K-12 . Restriction analysis of the clones demonstrated that about 15 kilobases were common to all clones and a further 5 kilobases were common to the Ogawa clones . The O antigens expressed by E . coli K-12 had the specificity of V . cholerae . Antibodies raised against E . coli K-12 that harbor one of these clones, pPM1001 (Inaba), were as highly protective in the infant mouse model system as were antibodies to V . cholerae itself . Introduction of such clones into suitable carrier strains could be expected to produce a good oral immunogen against cholera.

Biochim Biophys Acta, 1986 Jul 23, 850(3), 466 - 72
The sodium cycle . III . Vibrio alginolyticus resembles Vibrio cholerae and some other vibriones by flagellar motor and ribosomal 5S-RNA structures; Bakeeva LE et al.; An electron microscopic study of the basal bodies of the Vibrio albinolyticus flagellum revealed a four-disc structure . The diameters of the two discs localized closer to the cytoplasmic membrane proved to be about 2-fold shorter than those of the two others . In this respect the basal body of V . alginolyticus resembles very much that of V . cholerae described by Ferris and co-workers . The sequence of the V . alginolyticus ribosomal 5S-RNA showed that it is similar to those of V . cholerae, V . harveyi and some other vibriones . On the basis of the 5S-RNA sequences, a dendrogram of prokaryotes is presented . It confirmed the suggestion that V . alginolyticus is a typical representative of Vibrionaceae rather than a 'monster' greatly differing from other vibriones . Possible evolutionary relation of various bacterial species possessing the primary Na+ pumps is discussed.

Biochim Biophys Acta, 1986 Jul 23, 850(3), 458 - 65
The sodium cycle . II . Na+-coupled oxidative phosphorylation in Vibrio alginolyticus cells; Dibrov PA et al.; The role of Na+ in Vibrio alginolyticus oxidative phosphorylation has been studied . It has been found that the addition of a respiratory substrate, lactate, to bacterial cells exhausted in endogenous pools of substrates and ATP has a strong stimulating effect on oxygen consumption and ATP synthesis . Phosphorylation is found to be sensitive to anaerobiosis as well as to HQNO, an agent inhibiting the Na+-motive respiratory chain of V . alginolyticus . Na+ loaded cells incubated in a K+ or Li+ medium fail to synthesize ATP in response to lactate addition . The addition of Na+ at a concentration comparable to that inside the cell is shown to abolish the inhibiting effect of the high intracellular Na+ level . Neither lactate oxidation nor delta psi generation coupled with this oxidation is increased by external Na+ in the Na+-loaded cells . It is concluded that oxidative ATP synthesis in V . alginolyticus cells is inhibited by the artificially imposed reverse delta pNa, i.e., {Na+}in greater than {Na+}out . Oxidative phosphorylation is resistant to a protonophorous uncoupler (0.1 mM CCCP) in the K+-loaded cells incubated in a high Na+ medium, i.e., when delta pNa of the proper direction {( Na+}in less than {Na+}out) is present . The addition of monensin in the presence of CCCP completely arrests the ATP synthesis . Monensin without CCCP is ineffective . Oxidative phosphorylation in the same cells incubated in a high K+ medium (delta pNa is low) is decreased by CCCP even without monensin . Artificial formation of delta pNa by adding 0.25 M NaCl to the K+-loaded cells (Na+ pulse) results in a temporary increase in the ATP level which spontaneously decreases again within a few minutes . Na+ pulse-induced ATP synthesis is completely abolished by monensin and is resistant to CCCP, valinomycin and HQNO . 0.05 M NaCl increases the ATP level only slightly . Thus, V . alginolyticus cells at alkaline pH represent the first example of an oxidative phosphorylation system which uses Na+ instead of H+ as the coupling ion.

Biochim Biophys Acta, 1986 Jul 23, 850(3), 449 - 57
The sodium cycle . I . Na+-dependent motility and modes of membrane energization in the marine alkalotolerant vibrio Alginolyticus; Dibrov PA et al.; Respiration, membrane potential generation and motility of the marine alkalotolerant Vibrio alginolyticus were studied . Subbacterial vesicles competent in NADH oxidation and delta psi generation were obtained . The rate of NADH oxidation by the vesicles was stimulated by Na+ in a fashion specifically sensitive to submicromolar HQNO (2-heptyl-4-hydroxyquinoline N-oxide) concentrations . The same amounts of HQNO completely suppressed the delta psi generation . Delta psi was also inhibited by cyanide, gramicidin D and by CCCP + monensin . CCCP (carbonyl cyanide m-chlorophenylhydrazone) added without monensin exerted a much weaker effect on delta psi . Na+ was required to couple NADH oxidation with delta psi generation . These findings are in agreement with the data of Tokuda and Unemoto on Na+-motive NADH oxidase in V . alginolyticus . Motility of V . alginolyticus cells was shown to be (i) Na+-dependent, (ii) sensitive to CCCP + monensin combination, whereas CCCP and monensin, added separately, failed to paralyze the cells, (iii) sensitive to combined treatment by HQNO, cyanide or anaerobiosis and arsenate, whereas inhibition of respiration without arsenate resulted only in a partial suppression of motility . Artificially imposed delta pNa, i.e., addition of NaCl to the K+ -loaded cells paralyzed by HQNO + arsenate, was shown to initiate motility which persisted for several minutes . Monensin completely abolished the NaCl effect . Under the same conditions, respiration-supported motility was only slightly lowered by monensin . The artificially-imposed delta pH, i.e., acidification of the medium from pH 8.6 to 6.5 failed to activate motility . It is concluded that delta mu Na+ produced by (i) the respiratory chain and (ii) an arsenate-sensitive anaerobic mechanism (presumably by glycolysis + Na+ ATPase) can be consumed by an Na+ -motor responsible for motility of V . alginolyticus.

Brain Res, 1986 Jul 16, 378(1), 120 - 6
Evidence that ruthenium red disturbs the synaptic transmission in the rat hippocampal slices through interacting with sialic acid residues; Wieraszko A; Ruthenium red (RR) at a concentration of 0.71 mM selectively blocked synaptic transmission in hippocampal slices . Antidromically evoked potentials and fibre potentials were only little affected . The action of RR was reversible by washout, but only following shorter (40-50 min) times of incubation . After longer incubation times (hours), the abolished population spike did not recover after washout but could be restored by facilitation of the calcium transport into the nerve terminal with 3,4-diaminopyridine . Partial liberation of sialic acid with neuraminidase from Vibrio Cholerae markedly increased the time after which the potential was abolished by RR . Exogenously added gangliosides and sialic acid also delayed the action of RR . Calcium at a concentration of 13.2 mM prevented or reduced the RR effect . It is concluded that RR binds to sialic acid residues, interfering with neurotransmission by disturbing the calcium transport into the cell.

Appl Environ Microbiol, 1986 Jul, 52(1), 211 - 3
Plasmid carriage in Vibrio vulnificus and other lactose-fermenting marine vibrios; Davidson LS et al.; A total of 42 clinical and environmental isolates of Vibrio vulnificus were examined for plasmid carriage . Of these, only five (12%) harbored plasmids, which were of various molecular weights . In contrast, 20 of 32 (62.5%) unidentified lactose-fermenting Vibrio spp . were found to possess plasmids with masses of 2.1 to 150 megadaltons . In these isolates, multiple plasmids were common, with an average of 2.25 plasmids per plasmid-containing strain . Attempts to demonstrate a correlation with the plasmids identified in the various Vibrio spp . and a variety of phenotypic traits, production of several enzymes potentially involved in virulence, cytotoxicity for Chinese hamster ovary cells, and mouse lethality were unsuccessful . A correlation was observed, however, between the presence of a 6.5-megadalton plasmid and resistance to pteridine 0/129 . It was concluded that V . vulnificus, unlike most other Vibrio spp., shows a general lack of these extrachromosomal elements.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Jul, (7), 21 - 4
{2-cycle culture of Vibrio cholerae in a fermenter}; Nenkov PKh et al.; The method for the two-cycle cultivation of V . cholerae, reference strains Inaba and Ogawa, in a fermenter has been developed . In the first and second cultivation cycles no difference in the yield of the biomass and in the ultrastructure of the vibrios has been established . In the second cycle a decrease in the minimum time of generation has been observed . The vaccines obtained in the first and second cultivation cycles have not been found to differ in their antigenic and immunogenic activity . The vaccines meet the WHO requirements.

Arch Dermatol, 1986 Jul, 122(7), 818 - 20
Hemorrhagic bullae associated with Vibrio vulnificus septicemia . Report of two cases; Tyring SK et al.; Bullous lesions associated with Vibrio vulnificus infection developed in two patients, both of whom had hepatic cirrhosis . One patient had a recent history of ingestion of raw oysters, while the other patient had recently exposed skin lacerations to sea water . Both patients died within 24 hours of hospitalization, in spite of antibiotic treatment . Vibrio vulnificus was isolated from blood and bullae in both patients . Histologic examination of skin biopsy specimens revealed epidermal/dermal separation and clusters of bacteria within dermal vessels with a negligible inflammatory response.

Appl Environ Microbiol, 1986 Jul, 52(1), 142 - 5
Chitinase determinants of Vibrio vulnificus: gene cloning and applications of a chitinase probe; Wortman AT et al.; To initiate study of the genetic control of chitinolytic activity in vibrios, the chitobiase gene was isolated by cloning chromosomal DNA prepared from Vibrio vulnificus . Chimeric plasmids were constructed from Sau3A I partial digests of chromosomal DNA by ligating 5 to 15-kilobase fragments into the BamHI site, i.e., in the Tcr gene, of pBR322 (Amr Tcr) . The resulting plasmids were transformed into Escherichia coli DH1 . Chitobiase activity of the insert-bearing clones was detected by using a chromogenic substrate, p-nitrophenyl-N-acetyl-beta, D-glucosaminide, and confirmed by the appearance of a fluorescent end product from the hydrolysis of 4-methylumbelliferyl-beta,D-N-N'-diacetylchitobiose . Endochitinase activity was demonstrated by liberation of water-soluble products produced by the degradation of {3H}chitin . Transformation of E . coli Y10R (lacY) with plasmids from chitinase-positive clones restored the lactose-positive phenotype, suggesting the presence of a permease associated with chitinase activity . Physical mapping of plasmids containing the chitinase determinants indicate that transcription of these genes in E . coli may be initiated at a V . vulnificus promoter.

J Bacteriol, 1986 Jul, 167(1), 375 - 8
Isolation and characterization of the Vibrio cholerae recA gene; Hamood AN et al.; A 3.6-kilobase PstI fragment was isolated from a Vibrio cholerae chromosomal DNA library and shown to encode RecA-like activity in complementation studies with Escherichia coli recA mutants . Although DNA hybridization experiments failed to detect any homology between the E . coli and V . cholerae recA genes, hyperimmune antiserum produced against purified E . coli RecA protein recognized epitopes shared by the V . cholerae protein . The V . cholerae chromosomal fragments, when cloned and transferred to E . coli, provided the missing recA functions, including resistance to the alkylating agent methyl methanesulfonate, resistance to UV irradiation, and promotion of homologous recombination in Hfr mating experiments.

Diagn Microbiol Infect Dis, 1986 Jul, 5(2), 99 - 111
Production of extracellular enzymes and cytotoxicity by Vibrio vulnificus; Oliver JD et al.; Thirty-three strains of Vibrio vulnificus of clinical and environmental origin were examined for production of 12 extracellular enzymes of potential importance to the virulence of this bacterium . Strains of Vibrio vulnificus were consistent in their production of protease, mucinase, lipase, chondroitinase, hyaluronidase, DNase, sulfatase, and hemolysin . No differences between clinical and environmental isolates were noted . Although none of the enzymes appeared to correlate with the ability of these strains to produce lethality in mice, the production of hemolysin and of a protease with activity against native serum albumin may be significant in the pathogenesis of the potentially fatal infections produced by this organism . The production of several of these exoenzymes also appeared to correlate with pathogenicity in the seven other Vibrio species examined . Culture filtrates of all virulent strains of Vibrio vulnificus were cytotoxic for Chinese hamster ovary cells, whereas those of the strains of Vibrio parahaemolyticus and Vibrio alginolyticus examined lacked this activity.

J Bacteriol, 1986 Jul, 167(1), 411 - 4
Expression of luciferases from Vibrio harveyi and Vibrio fischeri in filamentous cyanobacteria; Schmetterer G et al.; Shuttle vectors that had previously been shown to replicate both in Escherichia coli and in strains of Anabaena spp . were used to transfer the lux genes from Vibrio harveyi and Vibrio fischeri into Anabaena spp . The level of expression of luciferase in the cyanobacteria (up to 7,000 quanta cell-1 s-1) makes these genes good candidates for use as promoter probes during the differentiation of certain cells in a filament into heterocysts.

J Bacteriol, 1986 Jul, 167(1), 57 - 65
Characterization of anguibactin, a novel siderophore from Vibrio anguillarum 775(pJM1); Actis LA et al.; Anguibactin, a siderophore produced by cells of Vibrio anguillarum 775 harboring the pJM1 plasmid, has now been isolated from the supernatants of iron-deficient cultures . This iron-reactive material was purified by adsorption onto an XAD-7 resin and subsequent gel filtration on a Sephadex LH-20 column . The resulting neutral compound produced an ion at m/z 348 in mass spectrometry and contained one sulfur, four oxygen, and four nitrogen atoms as determined by elemental analysis . Its strong UV absorbance and blue fluorescence were suggestive of a phenolic moiety . In colorimetric reactions anguibactin behaved like a catechol . The catechol assignment was supported by the appearance of a new absorption band at 510 nm in the ferric complex and by the appearance of peaks at 1,367, 1,447, 1,469, and 1,538 cm-1 in the resonance Raman spectrum . In addition, the infrared spectrum gave evidence of a secondary amide function, but no free carboxylic acid or hydroxamic acid groups were observed . A third iron-ligating group was suggested by the liberation of three protons during iron binding; mass spectrometry of the resulting material yielded a molecular ion characteristic of a 1:1 complex of ferric anguibactin . The purified anguibactin exhibited specific growth-promoting activity under iron-limiting conditions for a siderophore-deficient mutant of V . anguillarum 775(pJM1) . A novel structure for anguibactin was indicated by the failure of a large number of known siderophores and synthetic chelators to yield a similar type of specific cross-feeding in the V . anguillarum bioassay.

J Bacteriol, 1986 Jul, 167(1), 210 - 8
Regulation of lateral flagella gene transcription in Vibrio parahaemolyticus; Belas R et al.; Two distinctly different organelles of locomotion are produced by Vibrio parahaemolyticus . The polar flagellum is responsible for motility in a liquid environment (swimming), and the lateral flagella enable the bacteria to move over surfaces (swarming) . Synthesis of lateral flagella occurs when V . parahaemolyticus is grown on agar media but not when it is grown in liquid media . We used lux (luminescence gene) fusions to conveniently and sensitively analyze the factors which influence transcription of lateral flagella genes (laf) . Transposon mini-Mu lux was used to mutagenize V . parahaemolyticus and to generate laf::lux transcriptional fusions . Mutants with insertions of mini-Mu lux in laf genes were defective in the swarming phenotype and produced light when the bacteria were propagated on agar media, but not when cells were grown in liquid media . Thus, surface-dependent expression of lateral flagella synthesis is controlled by regulation of transcription . Such fusion strains were also used to further define the environmental conditions which induce laf gene expression . Cultivation on media solidified by gelling agents other than agar also induced light production in fusion strains, as did growth on a variety of hydrophilic membrane filters suspended over liquid media . Growth at an air-surface interface was not necessary for expression since embedding the fusion strains in agar was also effective . Furthermore, induction of laf gene transcription could also be accomplished by increasing the viscosity of the liquid medium by the addition of a high-molecular-weight polymer such as polyvinylpyrrolidone . Increase in luminescence of the fusion strains was detected within 30 min of initiation of the inducing circumstance, and reversal of induction, e.g., by dilution of the viscous medium, resulted in a rapid decline in the rate of increase in luminescence . Conditions that induced luminescence in the fusion strains also induced the synthesis of lateral flagella in wild-type V . parahaemolyticus . The growth environment of the genes, and it appears that the signal that triggers laf expression is physical rather than chemical in nature . Possibilities for a sensing mechanism are discussed.

Hum Nutr Clin Nutr, 1986 Jul, 40(4), 249 - 54
Effect of boiled-rice feeding in childhood cholera on clinical outcome; Khin-Maung-U et al.; Forty-eight children, aged 2-5 years, presenting with watery diarrhoea of less than 48 h duration at home prior to hospitalization, were admitted into a randomized controlled clinical trial, 24 children being treated during the first 24 h of admission with oral rehydration solution (ORS) alone and 24 children being given 'ORS plus boiled-rice feeding' . The latter group received boiled-rice to supply at least 55 kcal/kg/d (about 150 g boiled-rice per feed, given four times daily) . Vibrio cholerae were isolated by stool culture on admission from all children . No antibiotics were given . Clinical characteristics of children in the two treatment groups were comparable . Among children given 'ORS plus boiled rice', there was a significant increase in volume of diarrhoea stools (P less than 0.05), duration of diarrhoea in hospital (P less than 0.01), and more frequent diarrhoea motions (not significant statistically) . However, the children fed boiled rice absorbed and retained 176 ml more fluid, and had gain in body weight comparable to that observed in children who were not fed during the first 24 h of hospitalization.

Carbohydr Res, 1986 Jul 1, 149(2), 293 - 8
The effect of removal of D-fructose on the antigenicity of the lipopolysaccharide from a rough mutant of Vibrio cholerae Ogawa; Kaca W et al.; The lipopolysaccharide (LPS) of a rough mutant (95R) of Vibrio cholerae Ogawa has been investigated chemically and serologically . D-Fructose was released from LPS under conditions (10mM trifluoroacetic acid, 60 degrees, 1 h) that liberated no other sugar constituent of the LPS (2-amino-2-deoxy-D-glucose, D-glucose, L-glycero-D-manno-heptose) . Upon periodate oxidation, D-fructose and D-glucose were oxidised quantitatively, whereas approximately 50% of heptose was periodate-resistant . The data indicate that D-fructose does not link the polysaccharide and lipid A portion as proposed earlier, and suggest that D-fructose is present as a branch . By passive hemolysis inhibition, it was shown that the release of D-fructose paralleled the exposure of an antigenic determinant cryptic in LPS.

Schweiz Med Wochenschr, 1986 Jun 21, 116(25), 856 - 8
{Cholera in Fribourg . Apropos a case}; Nicole A et al.; A 41-year-old man from Cameroon was admitted to the Hospital of Fribourg (Switzerland) for watery diarrhea 3 months after cholera vaccination . Stool cultures were positive for Vibrio cholerae, biotype El Tor . Treatment consisted of rehydration (up to 9 litres per day) and correction of metabolic acidosis . No antibiotics were administered . The diagnosis of cholera should be considered in the presence of watery diarrhea in travellers or immigrants from countries suffering from the pandemic, even though, before this case, there had been no registered case for four years in Switzerland and none for twenty years in Fribourg.

Vet Immunol Immunopathol, 1986 Jun, 12(1-4), 83 - 92
Migration of fish leucocytes in vitro: the effect of factors which may be involved in mediating inflammation; Nash KA et al.; Plaice (Pleuronectes platessa L.) neutrophils were isolated from the kidney on a discontinuous Percoll gradient and from the peritoneal cavity at the peak of a glycogen-elicited inflammatory response . The migratory ability of neutrophils was assessed using a 48-well microchemotaxis chamber, with an incubation of 1.5 h at 12 degrees C . The two neutrophil populations showed different responses to N-formylmethionyl-leucyl-phenylalanine (FMLP) . Whereas kidney neutrophils only showed a significant enhancement of migration at 10(-7) M, inflammatory neutrophils exhibited a bimodal response, with one peak of migratory activity at 10(-9) M and a second at greater than 10(-6) M . Kidney neutrophils showed a consistent response with various concentrations of a 24 h culture supernatant of Vibrio alginolyticus . In every case increased migration was observed with 5-, 10- and 100-fold dilutions, with the latter two conditions producing a significant enhancement (p less than 0.01 and p less than 0.05 respectively) . The undiluted and 2-fold diluted supernatant caused a decreased cell migration compared with control values . The supernatant from kidney neutrophils cultured with serum-opsonized, heat-killed V . alginolyticus produced greater migratory activity than neutrophils or the treated bacteria incubated alone (the controls) . In each case, the enhanced activity of the supernatant was detectable by 1 h of incubation . By 4 h, the activity of the neutrophil/bacteria supernatant was significantly higher than that of the controls (p less than 0.01), but by 24 h had fallen to control levels . There was no evidence for a chemotactic response with FMLP, the bacterial supernatant or the neutrophil-derived factor and the responses were therefore assumed to be chemokinetic.

Vet Immunol Immunopathol, 1986 Jun, 12(1-4), 243 - 50
Effects of metals on the chemiluminescent response of rainbow trout (Salmo gairdneri) phagocytes; Elsasser MS et al.; Quantification of an induced chemiluminescent (CL) response in phagocytes is currently being evaluated as an indicator system for determining those environmental pollutants that may predispose fish to disease . A CL assay was developed using phagocytes from the pronephros of rainbow trout (Salmo gairdneri) . The CL response of phagocytes to phorbol myristate acetate, a chemical inducer of CL, was shown to be dose-dependent . The response to five species of bacteria was also evaluated . Staphylococcus aureus and Aeromonas hydrophila produced the most intense CL responses and the longest duration of response (100 min.) Yersinia ruckeri induced an immediate strong CL response of short duration (20 min.) whereas Vibrio anguillarum and Aerococcus viridans failed to stimulate CL under the test conditions employed . The effect of sub-toxic levels of Cu, Al, and Cd on the CL response of phagocytes to S . aureus was examined using phagocytes exposed to the metals immediately before assay or after 1 hr or 24 hr exposure times . Copper caused a significant decrease in CL to the baseline level under all treatment conditions upon stimulation with S . aureus . Similar results were obtained with Al except that the decrease in CL, although significant, was not to the baseline level . In contrast, Cd caused a significant increase in CL when added 1 hr prior to or immediately before the assay; but, following a 24 hr exposure, the results were variable, in that either no change or a decrease was observed . The addition of Cu to phagocytes already exhibiting a strong CL response to S . aureus caused an immediate decrease in CL to that seen with the negative controls.

Appl Environ Microbiol, 1986 Jun, 51(6), 1216 - 9
Isolation of non-O1 Vibrio cholerae serovars from surface waters in western Colorado; Rhodes JB et al.; Non-O1 Vibrio cholerae was isolated from rivers, creeks, washes, irrigation canals, and ditches in western Colorado during the summer of 1985 . The organism occurred in fresh water (less than or equal to 5 mmol of Na+ per liter) as well as in water of higher salinity (approximately equal to 17 mmol per liter) . Sixteen serovars of non-O1 V . cholerae were Sixteen serovars of non-O1 V . cholerae were identified among the environmental isolates . All of the isolates were cytotoxic to Y-1 mouse adrenal cells.

J Clin Microbiol, 1986 Jun, 23(6), 1091 - 5
Synthetic oligodeoxyribonucleotide probes to detect Kanagawa phenomenon-positive Vibrio parahaemolyticus; Nishibuchi M et al.; Synthetic oligodeoxyribonucleotide probes were used in the colony hybridization test to examine the association between the Kanagawa phenomenon (KP) and the thermostable direct hemolysin gene (tdh) of Vibrio parahaemolyticus . Representative V . parahaemolyticus strains with a variety of KP reactions and 17 other Vibrio species were examined for homology with four synthetic oligodeoxyribonucleotide probes (19 to 21 bases long) representing different regions of the tdh structural gene . Under stringent conditions, two of the probes were capable of distinguishing KP-positive V . parahaemolyticus from KP-negative or KP weak-positive V . parahaemolyticus which possesses mutated tdh genes . Vibrio hollisae strains hybridized with all four probes under reduced stringency, suggesting that they have tdh-related genes which are homologous but not identical to the tdh gene in all the regions examined . The results suggest that the colony hybridization test with the synthetic oligonucleotide probes is more suitable for the definitive determination of KP-positive strains than the hybridization with the larger gene probe or immunological assays.

Vet Immunol Immunopathol, 1986 Jun, 12(1-4), 39 - 45
Salmonid B lymphocytes demonstrate organ dependent functional heterogeneity; Irwin MJ et al.; The passive hemolytic plaque assay was used to examine the functional heterogeneity of antibody producing cells in salmonid immune organs . In this study, the antibody response to Vibrio anguillarum antigens was induced by the injection of a somatic antigen extract . This antigen was also coated onto sheep red blood cells (SRBC) for plaque forming cell (PFC) determination . Previous studies have demonstrated that this response is antibody dependent and antigen specific (Kaattari and Irwin, 1985) . The present study was focused upon the heterogeneity of antibody producing cells that arise in the spleen, anterior and posterior kidney of immunized coho salmon (Oncorhynchus kisutch) . The functional heterogeneity of lymphocytes was assessed by histogram analysis of the antigen inhibition profiles of the plaque forming responses . These analyses have revealed that the anterior kidney lymphocytes possess a much more restricted profile of antibody specificities than do lymphocytes from the posterior kidney or spleen . These data suggest that B cell repetoires differ among the immune organs of salmonids.

Carbohydr Res, 1986 Jun 1, 149(1), 59 - 64
Reconstitution of the masking effect of sialic acid groups on sialidase-treated erythrocytes by the action of sialyltransferases; Kelm S et al.; Glutardialdehyde-fixed or native rat erythrocytes were partially desialylated by the action of Vibrio cholerae sialidase, resulting in the binding of these cells to homologous peritoneal macrophages . Resialylation of these erythrocytes by purified alpha-(2----3)- or alpha-(2----6)-sialyltransferases with CMP-N-acetylneuraminic acid led to the incorporation of 60-80% of the enzymically released sialic acid . Binding of the resialylated erythrocytes to peritoneal macrophages was reduced when compared with corresponding, partially desialylated erythrocytes . Thus, the amount of transferred sialic acid was sufficient to demonstrate reconstitution of the masking effect of sialic acids.

Microb Pathog, 1986 Jun, 1(3), 283 - 8
Expression of the Escherichia coli lamB gene in Vibrio cholerae; Harkki A et al.; A phage lambda mediated transduction system was devised to facilitate molecular analysis of Vibrio cholerae . A lamB expression plasmid, pAMH62 was introduced into Vibrio cholerae by conjugation . The resulting V . cholerae derivatives harboring pAMH62 produced substantial amounts of the LamB protein . This protein was properly inserted into the outer membrane, as suggested by (i) its localization into the cell envelope, (ii) its association with the peptidoglycan layer of the cell wall, and (iii) its function as receptor for phage lambda . In vivo packaged cosmids were efficiently transduced into these strains of V . cholerae.

J Gen Microbiol, 1986 Jun, 132 ( Pt 6), 1641 - 6
Immunochemical properties of the lipopolysaccharide O-antigen of Vibrio cholerae O1 in relation to its chemical structure; Guhathakurta B et al.; D-Glucuronic acid and D-glucosamine have an immunodominant role in the lipopolysaccharide (LPS) O-antigen of both the Ogawa and the Inaba subtypes of Vibrio cholerae O1 . This was evident from the pronounced inhibitory effect on the LPS precipitin reaction demonstrated by these monosaccharides and by oligosaccharides containing either of them which were isolated from LPS hydrolysate . There was a considerable decrease in the antibody-combining capacity of chemically modified LPS in which the carboxyl group of the glucuronic acid had been reduced . Similarly, on deamination, the O-specific polysaccharide fraction of the LPS molecule from both subtypes completely lost the ability to precipitate the LPS antibody.

J Infect Dis, 1986 Jun, 153(6), 1108 - 18
M cell transport of Vibrio cholerae from the intestinal lumen into Peyer's patches: a mechanism for antigen sampling and for microbial transepithelial migration; Owen RL et al.; Viable Vibrio cholerae O1 were inoculated into the intestinal lumen of nonimmune rabbits . The vibrios were phagocytosed by M cells over Peyer's patch lymphoid follicles, carried in vesicles through the epithelium, and discharged among underlying lymphocytes and macrophages . Autoradiography of V . cholerae labeled with {2-3H}adenine confirmed transport . Indigenous bacteria with and without capsules were also taken up from control loops and carried through M cells into Peyer's patches . V . cholerae killed by acidification, formalin, heat, or UV irradiation were not taken up, a result that may have relevance for development of oral vaccines . Ruthenium red stain revealed gaps in the layer of mucus over M cells, glycocalyx bridging the space between vibrios and M cell microvilli, and knobby projections over membranes of M cell microvilli; these projections were not found over absorptive cells . M cells thus convey viable enteric microbes, including V . cholerae that are not otherwise invasive, into intestinal lymphoid tissue, where mucosal immune responses are initiated . Uptake and transport by M cells may also assist certain pathogenic bacteria in traversing the mucosal barrier and initiating systemic infection.

Biochem Biophys Res Commun, 1986 May 14, 136(3), 1030 - 5
N-ethylmaleimide desensitizes pH-dependence of K+/H+ antiporter in a marine bacterium, Vibrio alginolyticus; Nakamura T et al.; The K+/H+ antiporter of a marine bacterium, Vibrio alginolyticus, is strongly dependent upon the cytoplasmic pH and functions only at an internal pH above 7.7 . In alkaline buffer with an outwardly directed chemical gradient of K+ (delta pK), the internal pH was maintained at about 7.7 . Addition of N-ethylmaleimide (NEM) released cellular K+ and acidified the cytosol below pH 7.7 . The NEM effect was reversed by the addition of 2-mercaptoethanol: K+ efflux ceased, and the internal pH returned to about 7.7 . In acidic buffer, the internal pH was also regulated at about 7.6 even in the absence of delta pK . Following addition of NEM, the internal pH decreased below 7.6, dissipating delta pH . These results suggest that NEM desensitizes the pH-dependence of the K+/H+ antiporter, allowing the antiporter to function at an internal pH below 7.7.

Res Vet Sci, 1986 May, 40(3), 328 - 32
Phagocytic activity of macrophages of rainbow trout against Vibrio anguillarum and the opsonising effect of antibody and complement; Honda A et al.; The phagocytosis of Vibrio anguillarum by peritoneal macrophages from normal rainbow trout was enhanced by antibody and complement . Treatment of either macrophages or the bacteria by antibody also enhanced opsonisation . Five weeks after immunisation with V anguillarum, the phagocytic activity of macrophages from rainbow trout was increased significantly compared with the activity of those from unvaccinated fish . Although agglutinin titres did not increase until three weeks after immunisation, seven out of 10 fish challenged one week after immunisation survived, indicating that immunised fish had developed resistance to vibrio infection before significant levels of antibody or phagocytic activity became detectable.

Appl Environ Microbiol, 1986 May, 51(5), 1004 - 6
Virulence of Vibrio vulnificus strains from marine environments; Tison DL et al.; Vibrio vulnificus strains isolated from geographically diverse marine sources were compared with clinical isolates for phenotype and in vitro and in vivo production of virulence factors . There were no differences between environmental and clinical strains on the basis of biochemical characteristics or antimicrobial susceptibility patterns . Cytolysin and cytotoxin titers produced by environmental strains were generally comparable to those of clinical strains . Of 29 environmental isolates tested, 25 were pathogenic for mice . These data show that environmental V . vulnificus strains are phenotypically indistinguishable from clinical isolates and that approximately 90% of the environmental strains tested produced in vitro virulence factors and in vivo pathogenicity for mice comparable to those produced by clinical V . vulnificus isolates.

Pediatr Res, 1986 May, 20(5), 416 - 21
Human and bovine milk: comparison of ganglioside composition and enterotoxin-inhibitory activity; Laegreid A et al.; Milk gangliosides inhibit Vibrio cholerae enterotoxin and Escherichia coli heat-labile enterotoxin . Human milk gangliosides showed considerably higher enterotoxin-inhibitory activity compared to bovine and formula milk gangliosides as measured in vitro by enzyme-linked immunosorbent assay and in vivo in rabbit small bowel loops . While gangliosides from less than 1 ml human milk inhibited 0.1 microgram choleratoxin in vitro and in vivo, five to 10 times higher amounts of bovine milk gangliosides were necessary to achieve similar results . Analysis of the ganglioside composition in human, bovine, and bovine milk-based formula milk showed that the ganglioside patterns in human and bovine milk differed markedly . The ganglioside patterns of bovine milk and formula milk appeared identical . In human or bovine milk, the total amount of gangliosides was 11 mg/liter compared to 6 mg/liter in formula milk . The predominating ganglioside in human milk, monosialoganglioside 3 (74% of total gangliosides), was only a minor component (3%) of bovine milk gangliosides . Disialoganglioside 3 represented 80% of bovine milk gangliosides compared to 25% of the human milk gangliosides . Trace amounts of monosialoganglioside 1 were detected in human, as well as in bovine, milk by a sensitive high performance thin-layer chromatography immunoassay . The monosialoganglioside 1 content in human milk was 10 times higher than in bovine milk . We conclude that the higher nonimmunoglobulin enterotoxin-inhibitory activity in human milk compared to bovine milk is associated with the differences in the ganglioside fraction.

Am J Clin Pathol, 1986 May, 85(5), 644 - 6
Non-01 Vibrio cholerae bacteremia--complication of a LeVeen shunt; McCleskey FK et al.; Vibrio cholerae bacteremia occurred in a patient with cirrhosis after placement of a LeVeen shunt . At the time of bacteremia, cultures of peritoneal fluid were negative and fluid dynamics did not suggest spontaneous bacterial peritonitis . Despite apparent successful treatment of the bacteremia, relapse and death occurred with culture positivity of peritoneal fluid for V . cholerae . Simultaneously, blood cultures were positive for Klebsiella pneumoniae . Agglutination studies demonstrated the V . cholerae to be a non-01 strain . Insertion of a LeVeen shunt, which bypasses the hepatic clearance mechanisms, appeared to have allowed bacteremia to occur with this bacterium that is rarely isolated from blood . In patients with LeVeen shunts, bacteremia with noninvasive pathogens may occur, and in coastal areas, Vibrios should be considered when bacteremia occurs.

Infect Immun, 1986 May, 52(2), 476 - 83
Plasmid-mediated changes in virulence of Vibrio cholerae; Hamood AN et al.; The effects of several plasmids, including cloning vectors and R factors, on the virulence of Vibrio cholerae CA401R were determined by measuring the dose-related diarrheal response in orally challenged infant mice . The plasmids were also examined for their effects on the colonization ability of strain CA401R by joint infection experiments with a spectinomycin-resistant CA401 strain as an internal standard . One V . cholerae R factor, pVH2, enhanced the diarrheal response, while R factors Rts1 and pVH1 reduced it; plasmids RP4, pRK290, Sa, pSJ8, pSJ5, and pBR328 had no effect . The ability of the plasmids to affect in vitro toxin production by CA401R was variable . Cells containing large plasmids all showed a modest decrease in colonization ability . These results showed that some plasmids affected V . cholerae virulence, but that the cloning vectors pBR328, RP4, and pRK290 did not.

Brain Res, 1986 Apr 23, 371(2), 305 - 13
Evidence for the functional role of monosialoganglioside GM1 in synaptic transmission in the rat hippocampus; Wieraszko A et al.; The hippocampal slices were incubated with compounds which hydrolyze, modify or bind with sialic acid containing molecules . The efficiency of synaptic transmission was tested in the presence of these compounds . The size of the evoked extracellularly recorded potential following Schaffer collateral stimulation was used as an indicator of synaptic transmission efficiency . Sodium periodate (10 mM) and sodium perchlorate (59.2 mM) evoked a reversible (after washout) decrease in the size of the population spike . Higher concentration of sodium periodate (60 mM) abolished the size of the population spike, which was only poorly reversible after washout . Tetanus toxin, which binds to polysialogangliosides, and neuraminidase from Vibrio cholerae (an enzyme which splits off sialic acid from polysialogangliosides, leaving GM1 intact, and splits off sialic acid from sialoglycoproteins) had no influence on the size of the population spike . Cholera toxin, which binds to GM1, slightly reduced the size of the population spike . Incubation of the slices with neuraminidase from Arthrobacter ureafaciens (an enzyme which splits off sialic acid from all gangliosides, including GM1, and from sialoglycoproteins) abolished the population spike after 5 h . GM1 antiserum abolished the potential after approximately 100 min . The conclusion is drawn that of all gangliosides only GM1 is necessary to support synaptic transmission in Schaffer collateral-pyramidal cell synapses.

J Biol Chem, 1986 Apr 15, 261(11), 4805 - 11
Nucleotide sequence of the luxB gene of Vibrio harveyi and the complete amino acid sequence of the beta subunit of bacterial luciferase; Johnston TC et al.; The nucleotide sequence of the 1.30-kilobase EcoRI/BglII fragment from Vibrio harveyi carrying the majority of the luciferase beta subunit coding region (luxB gene) has been determined . The EcoRI/BglII fragment was derived from a 4.0-kilobase HindIII fragment carrying both luxA and luxB which was detected in a genomic clone bank based on the expression of bioluminescence from colonies of Escherichia coli carrying V . harveyi HindIII fragments in plasmid pBR322 (Baldwin, T . O., Berends, T., Bunch, T . A., Holzman, T . F., Rausch, S . K., Shamansky, L., Treat, M . L., and Ziegler, M . M . (1984) Biochemistry 23, 3663-3667) . The entire alpha subunit coding sequence (luxA gene) and the amino-terminal 13 codons of the beta subunit sequence (luxB gene) were contained on a 1.85-kilobase EcoRI fragment, the sequence of which has been reported (Cohn, D . H., Mileham, A . J., Simon, M . I., Nealson, K . H., Rausch, S . K., Bonam, D., and Baldwin, T . O . (1985) J . Biol . Chem . 260, 6139-6146) . The beta subunit coding sequence was found to terminate 972 bases past the start of the luxB coding sequence . The beta subunit had a calculated molecular weight of 36,349 and comprised a total of 324 amino acid residues; the alpha beta dimer had a molecular weight (alpha + beta) of 76,457 . There were 27 base pairs separating the stop codon of the beta subunit structural gene and a 340-base open reading frame extending to (and beyond) the distal BglII site . Approximately two-thirds of the beta subunit was sequenced by protein chemical techniques . The amino acid sequence predicted from the DNA sequence, with few exceptions, confirmed the chemically determined sequence, and the measured amino acid composition was in excellent agreement with the composition implied from the DNA sequence.

Biochem Biophys Res Commun, 1986 Apr 14, 136(1), 273 - 80
Effects of the Vibrio cholerae siderophore vibriobactin on the growth characteristics of L1210 cells; Bergeron RJ et al.; The microbial iron chelator vibriobactin, N-{3-(2,3-dihydroxybenzamido)propyl}-1,3-bis{2-(2, 3-dihydroxyphenyl)-trans-5-methyl-2-oxazoline-4-carboxamido}-propane, is shown to inhibit the growth of L1210 cells in culture, with an IC50 value of 2 microM . Its biological activity is assigned to the ligand's ability to chelate iron as indicated by the disappearance of its antimitotic properties on iron chelate preformation or on O-methylation . The ligand is shown to have pronounced effects on cell cycle kinetics introducing a G1/S phase block . When treated cells are washed free of the ligand with fresh media they cascade into a high S phase in 5 hrs . Furthermore, after exposure of L1210 cells to vibriobactin (10 microM) for 5 hrs followed by removal of the drug, cells display different doubling times relative to untreated controls, with lower bromodeoxyuridine (BrdUrd) incorporation and no apparent cell death as shown by a 51Cr release assay . The effects on cell kinetics are consistent with inhibition of ribonucleotide reductase . Finally, vibriobactin is also shown to be active with a human Burkitt lymphoma cell line (Daudi).

Infect Immun, 1986 Apr, 52(1), 45 - 9
Purification and characterization of Vibrio cholerae non-O1 heat-stable enterotoxin; Arita M et al.; A toxin which causes rapid fluid accumulation in a suckling mouse assay and which was produced by Vibrio cholerae non-O1 was investigated . The toxin was purified from the culture supernatant of V . cholerae non-O1 (strain A-5) by ammonium sulfate fractionation, hydroxyapatite treatment, ethanol extraction, column chromatographies on SP-Sephadex C-50 and DEAE-Sephadex A-25, and high-pressure liquid chromatography on a Lichrosorb RP-8 column . About 1.4 X 10(5)-fold purification was achieved, with a recovery of about 12% . Although the crude preparation was heat labile, the purified toxin was heat stable . The minimum effective dose of purified toxin was about 5 ng in the suckling mouse assay . The amino acid composition of the purified toxin was determined to be Asp(3), Glu(1), Ala(1), half-Cys(6), Ile(2), Leu(1), Phe(1), and Pro(1) . These data show the production of a new type of heat-stable enterotoxin (NAG-ST) by V . cholerae non-O1.

Infect Immun, 1986 Apr, 52(1), 319 - 22
Purification and partial characterization of a non-O1 Vibrio cholerae hemolysin that cross-reacts with thermostable direct hemolysin of Vibrio parahaemolyticus; Yoh M et al.; A newly identified hemolysin (NAG-rTDH), which is related to the thermostable direct hemolysin (Vp-TDH) of Vibrio parahaemolyticus produced by non-O1 Vibrio cholerae, was studied . NAG-rTDH was purified by successive column chromatographies on DEAE-cellulose and an immunoaffinity column coupled with anti-Vp-TDH immunoglobulin . The molecular weight of NAG-rTDH was estimated as 18,500, similar to that of Vp-TDH, as judged by sodium dodecyl sulfate slab gel electrophoresis, but its charge or molecular shape was different, judging from its electrophoretic mobility . The lytic activities of NAG-rTDH on erythrocytes of most animals were essentially similar to those of Vp-TDH, but that on sheep erythrocytes was different . The hemolytic activity of NAG-rTDH was stable on heating at 100 degrees C for 10 min, as was that of Vp-TDH . Immunological cross-reactivity between NAG-rTDH and Vp-TDH was demonstrated by both the Ouchterlony test and the neutralization test . Thus, we conclude that non-O1 V . cholerae produce a new type of hemolysin that is similar but not identical to the thermostable direct hemolysin of V . parahaemolyticus.

J Gen Microbiol, 1986 Apr, 132 ( Pt 4), 1027 - 33
Novel transmissible factors in a non-O1 Vibrio cholerae and a Vibrio sp; Smigocki AC et al.; Transmissible factors encoding production of lacunae (L factors) were demonstrated in a non-O1 Vibrio cholerae and a Vibrio sp . of recent environmental origin . Lacunae were produced in lawns of non-O1 V . cholerae indicator strains under the same assay conditions as those where lacunae were produced by the well characterized P fertility plasmid of V . cholerae O1 and the V fertility factor found in a non-cholera vibrio strain . The origin of the lacunae produced by strains harbouring the V and L factors was examined . No vibriocin or phage activity was found in culture supernates or in lacunae produced by the strains, suggesting that, as in the case of the P plasmid, the lacunae probably represent sites of active mating . Unlike the P plasmid, neither the Vn or L factor could be detected or isolated by conventional plasmid techniques.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Apr, 261(2), 232 - 9
Virulence mechanisms associated with clinical isolates of non-O1 Vibrio cholerae; Shehabi AA et al.; Twenty one isolates of non-O1 V . cholerae from patients with diarrheal illness were examined for the presence of potential virulence mechanisms . The motile strains (90%) produced cell-associated mannose-sensitive hemagglutinins which reacted with human group O, chicken, sheep and rabbit erythrocytes . Motile isolates also attached to embryonic intestinal epithelial cells (ATCC 407), and the adherence was not inhibited by the presence of 1% D-mannose . All vibrio isolates hemolyzed sheep erythrocytes . Three vibrio isolates (14%) harbored two or three plasmids which ranged in size between 1.7 and 5.2 megadaltons . The presence of the plasmid did not correlate with the presence of hemolysin, hemagglutinins, adhesions or antibiotic resistance in any of the isolates . Thus, it appears that multiple factors associated with bacterial cell surfaces influence adhesin and apparently pathogenic potential of the non-O1 vibrio isolates in the host intestine.

J Gen Microbiol, 1986 Apr, 132 ( Pt 4), 877 - 81
Determination of the spiral conformation of Aquaspirillum spp . by scanning electron microscopy of elongated cells induced by cephalexin treatment; Konishi H et al.; The effect of the beta-lactam antibiotic cephalexin on the spiral conformation of cells of Aquaspirillum spp . was examined by scanning electron microscopy . A . itersonii and A . peregrinum, which are known to have a left-handed spiral shape, elongated and still showed left-handed spirals in medium containing cephalexin . The spiral conformation of the elongated cells is therefore considered to represent the natural condition . The spiral conformations of A . metamorphum and A . psychrophilum grown in ordinary cultures were difficult to determine because they have short cells without a complete spiral . After cephalexin treatment, the cells of these species elongated and displayed spiral forms, right-handed in A . metamorphum and left-handed in A . psychrophilum . This elongation method may be useful for checking and determination of the spiral handedness of short spiral or curved bacteria such as vibrios.

Mol Gen Genet, 1986 Apr, 203(1), 58 - 63
Cloning and expression in Escherichia coli of a recA-like gene from Vibrio cholerae; Paul K et al.; A library containing more than 80% of the Vibrio cholerae genome was constructed by cloning BamH1 restriction fragments into pBR322 . Using interspecific complementation of an Escherichia coli recA mutant with plasmids containing the gene bank of V . cholerae, a recA-like gene was identified . The recombinant plasmid, designated as pDP145, contained a 1.45 kb segment of V . cholerae DNA which codes for a protein of molecular weight 39,000 . The product of this gene confers methyl methane sulphonate resistance on the E . coli recA mutant, suppresses its ultraviolet (UV) light sensitive phenotype and has proteolytic activity on the phage lambda repressor . Induction of a 39,000 dalton protein in UV-irradiated V . cholerae cells was demonstrated.

Infect Immun, 1986 Apr, 52(1), 279 - 84
Molecular cloning and expression in Escherichia coli K-12 of the gene for a hemagglutinin from Vibrio cholerae; Franzon VL et al.; Using antiserum to the purified soluble hemagglutinin we isolated an Escherichia coli K-12 clone expressing the gene for a hemagglutinin from Vibrio cholerae 569B . The plasmid present in this clone was designated pPM471 . By deletion analysis with both specific restriction endonucleases and Bal 31 nuclease, we localized the gene, to a 0.72-kilobase region of DNA, implying a molecular weight of less than 27,000 for the protein . Analysis in E . coli K-12 minicells of plasmids containing the cloned gene and deletion derivatives of these plasmids identified a protein of 24,000 daltons correlating with hemagglutinating activity . Using the cloned gene as a probe, we demonstrated the presence of homologous DNA in a variety of V . cholerae strains including both biotypes . Furthermore, by screening gene banks in E . coli K-12 of V . cholerae El Tor O17, we isolated several El Tor clones containing this region of DNA and also expressing hemagglutinating activity.

Med J Aust, 1986 Mar 3, 144(5), 229 - 34
Investigation of cholera acquired from the riverine environment in Queensland; Bourke AT et al.; Since February 1977, five patients with cholera apparently acquired the infection from the riverine environment in Queensland . A total of 13 rivers have now yielded at least one isolate of Vibrio cholerae 01 biovar El Tor . Investigations indicate that the organism, including toxigenic strains, can survive and multiply in the riverine environment . No human or animal reservoirs and no ecological niches were identified and no route of importation or dissemination of the organism was discovered . The microbiological examination of faeces in all medical laboratories in Australia should include methods for detecting the cholera organism as a routine . When confronted with a cholera infection, medical practitioners should obtain a history of recent travel, both in Australia and overseas.

Appl Environ Microbiol, 1986 Mar, 51(3), 593 - 7
Serotyping of Vibrio anguillarum; Sorensen UB et al.; A serotyping scheme based on the detection of O antigens by slide agglutination in fish-pathogenic strains of Vibrio anguillarum is presented . Over a period of 5 years 270 Vibrio strains from feral and cultured fish, 189 strains from the environment, and 36 strains from invertebrates were collected . The strains were divided into 10 distinct serotypes (O1 through O10) . More than 90% of the fish-pathogenic strains, but only 40% of the environmental strains, were typable; 71% of the strains isolated from cultured rainbow trout were serotype O1, whereas 78% of the strains isolated from feral fish were serotype O2 . No dominating environmental serotype was found . A serotyping system for V . anguillarum is proposed . A total of 90 strains received from culture collections and laboratories in different countries were typed according to the present system.

J Clin Microbiol, 1986 Mar, 23(3), 624 - 6
Persistence of cholera in the United States: isolation of Vibrio cholerae O1 from a patient with diarrhea in Maryland; Lin FY et al.; A case of cholera was identified in Baltimore County, Md., in October 1984 . The Vibrio cholerae O1 isolate from the patient was hemolytic, biotype El Tor, serotype Inaba, and was toxigenic by the Y-1 adrenal cell assay; on Southern blot analysis, the strain had a unique HindIII restriction site in the cholera toxin gene identical to that of other U.S . V . cholerae O1 isolates . Two days before he became ill, the patient had eaten meat from crabs harvested along the Texas coast.

J Clin Microbiol, 1986 Mar, 23(3), 411 - 5
Emergence of bactericidal and opsonizing antibody to Vibrio vulnificus following bacterial infection; Musher DM et al.; Virulent isolates of Vibrio vulnificus resist the bactericidal and opsonizing effects of normal human serum, in contrast to environmental isolates, which are highly serum susceptible . Immune responses to bacteremic V . vulnificus infections in human subjects have not been characterized . Serum from a patient who survived sepsis caused by V . vulnificus had substantial bactericidal and opsonizing immunoglobulin G (IgG) for his own bloodstream isolate . Killing was mediated by the classical complement pathway, whereas opsonization was effected by either the classical or the alternative pathway . IgG that reacted strongly with 55-, 58-, and 68-kilodalton outer membrane proteins was present in the patient's convalescent-phase serum but was absent from normal human serum . These findings suggest that humoral immunity to V . vulnificus, mediated by bactericidal and opsonizing antibody, emerges during infection and may be due, in part, to IgG directed against identifiable outer membrane proteins.

J Bacteriol, 1986 Mar, 165(3), 723 - 31
Effect of a recA mutation on cholera toxin gene amplification and deletion events; Goldberg I et al.; The cholera toxin operon (ctxAB) is located on a 7-kilobase pair variable genetic element which undergoes genetic duplication and amplification events in Vibrio cholerae . Amplification of the ctx genetic element was investigated by substituting the resident ctx loci of two V . cholerae strains with a DNA fragment encoding resistance to kanamycin . Although these strains were not normally resistant to greater than 150 micrograms of kanamycin per ml, spontaneous derivatives could be obtained that grew well on 3 mg of kanamycin per ml . Southern blot analysis of these highly resistant isolates demonstrated that the ctx element was amplified approximately 20-fold . This amplification process was completely inhibited in the absence of a functional recA gene . The V . cholerae RecA protein, therefore, is essential for cholera toxin gene amplification . Spontaneous deletions of the ctx structural genes were observed in both recA+ and recA- V . cholerae strains, although such deletions occurred at a 21-fold-lower frequency in the latter case . Structural analysis of these ctx amplification and deletion events supports a model for their formation that involves unequal crossing over between repetitive sequences located upstream and downstream of the ctx operon.

Infect Immun, 1986 Mar, 51(3), 964 - 5
Detection of anti-Vibrio vulnificus cytolysin antibodies in sera from mice and a human surviving V . vulnificus disease; Gray LD et al.; An enzyme-linked immunosorbent assay and a cytolysin neutralization assay were used detect anti-Vibrio vulnificus cytolysin antibodies in sera from mice and a human that survived V . vulnificus disease . The detection of antibodies against the cytolysin indicated that the cytolysin is produced in vivo, and this observation is consistent with the hypothesis that the cytolysin is involved in the pathogenesis of V . vulnificus disease.

Am J Epidemiol, 1986 Mar, 123(3), 424 - 30
Impact of epidemic cholera in a previously uninfected island population: evaluation of a new seroepidemiologic method; Harris JR et al.; During an investigation of a 1982 cholera outbreak in Truk, an area without endemic cholera, 254 post-outbreak serum specimens were collected from ill and well inhabitants of a single island . These were compared with 57 specimens collected in Truk in 1964, when heat-labile toxin-producing enterotoxigenic Escherichia coli was presumably endemic but cholera did not exist . The serum was tested for vibriocidal antibody and antitoxic antibodies to cholera toxin and heat-labile toxin and the ratio of the anti-cholera antitoxin to the anti-heat-labile antitoxin was calculated . The prevalences of elevated anti-cholera antitoxin and anti-heat-labile antitoxin levels were similar in both years: an elevated anti-cholera antitoxin was found in 30% of the 1964 and 80% of the 1982 specimens, while an elevated anti-heat-labile antitoxin was found in 40% (1964) and 84% (1982) . In contrast, vibriocidal antibody titers were elevated in none of the 1964 but in 64% of the 1982 specimens, and the anti-cholera/anti-heat-labile antitoxin ratio was elevated in 4% (1964) and 42% (1982) . The anti-cholera/anti-heat-labile antitoxin ratio appears to differentiate the antitoxic responses to Vibrio cholerae O1 and heat-labile toxin-producing enterotoxigenic E . coli, even in areas where both are prevalent, and should serve as a useful seroepidemiologic tool.

Genetika, 1986 Mar, 22(3), 399 - 405
{Genetic mapping of the regulatory gene determining the synthesis of enterotoxin in Vibrio cholerae}; Smirnova NI et al.; The data of genetic mapping of the cholera toxin regulatory gene by conjugation mating of Vibrio cholerae eltor donor strain with V . cholerae classica recipients are presented . The close genetic linkage of tox locus to pur-63 is shown . The gene order asp - cys - nal - pur-61 - trp - his - pur-63 - tox - ile of the chromosomal region examined is established.

Infect Immun, 1986 Mar, 51(3), 927 - 31
Identity of hemolysins produced by Vibrio cholerae non-O1 and V . cholerae O1, biotype El Tor; Yamamoto K et al.; Hemolysins purified from non-O1 Vibrio cholerae (non-O1 hemolysin) and a Vibrio cholerae O1, biotype El Tor (El Tor hemolysin) were investigated for their homology . The hemolysins were isolated from the culture supernatant fluids by ammonium sulfate precipitation and gel filtration on Sephadex G-100 columns . The purified hemolysins gave single bands with an identical mobility on conventional polyacrylamide gel disc electrophoresis . The molecular weights of the non-O1 and El Tor hemolysins were estimated to be about 60,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the amino acid compositions of the hemolysins were very similar . The specific activities of the hemolysins were identical, and both hemolysins were neutralized to the same extent with antisera against the homologous and heterologous hemolysins . Ouchterlony double immunodiffusion tests with both hemolysins and antihemolysin serum gave a common (fused) precipitin line . These data indicate that the non-O1 hemolysin is biologically, physicochemically, and immunologically indistinguishable from the El Tor hemolysin.

J Clin Microbiol, 1986 Mar, 23(3), 652 - 4
Molecular epidemiology of non-O1 Vibrio cholerae and Vibrio mimicus in the U.S . Gulf Coast region; Kaper JB et al.; Ten toxigenic Vibrio cholerae non-O1 and V . mimicus strains isolated from clinical and environmental sources in the U.S . Gulf Coast region were examined for genetic relatedness . Restriction digest patterns of chromosomal DNA and Southern blot analysis with a cholera toxin gene probe revealed that the strains exhibited greater genetic divergence than the highly conserved V . cholerae O1 strains isolated from clinical and sewage samples in this region.

J Bacteriol, 1986 Mar, 165(3), 715 - 22
Cloning of the Vibrio cholerae recA gene and construction of a Vibrio cholerae recA mutant; Goldberg I et al.; A recombinant plasmid carrying the recA gene of Vibrio cholerae was isolated from a V . cholerae genomic library, using complementation in Escherichia coli . The plasmid complements a recA mutation in E . coli for both resistance to the DNA-damaging agent methyl methanesulfonate and recombinational activity in bacteriophage P1 transductions . After determining the approximate location of the recA gene on the cloned DNA fragment, we constructed a defined recA mutation by filling in an XbaI site located within the gene . The 4-base pair insertion resulted in a truncated RecA protein as determined by minicell analysis . The mutation was spontaneously recombined onto the chromosome of a derivative of V . cholerae strain P27459 by screening for methyl methanesulfonate-sensitive variants . Southern blot analysis confirmed the presence of the inactivated XbaI site in the chromosome of DNA isolated from one of these methyl methanesulfonate-sensitive colonies . The recA V . cholerae strain was considerably more sensitive to UV light than its parent, was impaired in homologous recombination, and was deficient in induction of a temperate vibriophage upon exposure to UV light . We conclude that the V . cholerae RecA protein has activities which are analogous to those described for the RecA protein of E . coli.

J Hyg (Lond), 1986 Feb, 96(1), 49 - 57
The impact of physico-chemical stress on the toxigenicity of Vibrio cholerae; Miller CJ et al.; The expression of toxigenicity by Vibrio cholerae, before and after exposure to various conditions of salinity, pH and cation composition and concentration, has been measured . Exposure to these conditions did not select for hyper- or hypo-toxigenic strains . This suggests that toxigenic V . cholerae O1 are unlikely to lose their toxigenicity when exposed to environmental stress and that V . cholerae toxin production is not a response to the stresses included in this study . These results are consistent with an aquatic reservoir for toxigenic V . cholerae O1.

Arch Biochem Biophys, 1986 Feb 1, 244(2), 766 - 72
An NADH:quinone oxidoreductase of the halotolerant bacterium Ba1 is specifically dependent on sodium ions; Ken-Dror S et al.; The rate of NADH oxidation by inverted membrane vesicles prepared from the halotolerant bacterium Ba1 of the Dead Sea is increased specifically by sodium ions, as observed earlier in whole cells . The site of this sodium effect is identified as the NADH: quinone oxidoreductase, similarly to the other such system known, Vibrio alginolyticus (H . Tokuda and T . Unemoto (1984) J . Biol . Chem . 259, 7785-7790) . Sodium accelerates quinone reduction severalfold, but oxidation of the quinol, with oxygen as terminal electron acceptor, is unaffected . The sodium-dependent pathway of quinone reduction exhibits higher apparent affinity to extraneous quinone (Q-2) than the sodium-insensitive pathway, and is specifically inhibited by 2-heptyl-4-hydroxyquinoline N-oxide . ESR spectra of the membranes contain a feature at g = 1.98 which is tentatively identified as one originating from semiquinone . This feature is increased by NADH and decreased by addition of Na+, suggesting that, as proposed from different kinds of evidence for the V . alginolyticus system, sodium affects the semiquinone reduction step . As in the other system, the site of sodium stimulation in Ba1 probably corresponds to the site of sodium translocation, which was shown earlier (S . Ken-Dror, R . Shnaiderman, and Y . Avi-Dor (1984) Arch . Biochem . Biophys . 229, 640-649) to be linked directly to a redox reaction in the respiratory chain.

Am J Med, 1986 Feb, 80(2), 336 - 8
Gastroenteritis in patients with stool isolates of Vibrio vulnificus; Johnston JM et al.; Vibrio vulnificus, a marine Vibrio associated with severe extraintestinal infections, has not been previously implicated as a cause of infectious diarrhea . Three patients were identified with diarrheal illness from whom this organism was the sole pathogen recovered from cultured stool specimens . All three had eaten raw oysters within one week of becoming ill . These patients were all taking medication that reduces gastric acidity, two were heavy drinkers of alcohol, and one had unrecognized colon cancer; these factors may have predisposed to the development of disease . Clinicians should consider that V . vulnificus may be a cause of gastroenteritis in patients who have consumed raw oysters.

J Bacteriol, 1986 Feb, 165(2), 461 - 6
Luciferase-dependent oxygen consumption by bioluminescent vibrios; Makemson JC; Oxygen uptake due to luciferase in two luminous Vibrio species was estimated in vivo by utilizing inhibitors having specificities for luciferase (decanol) and cytochromes (cyanide) . Cyanide titration of respiration revealed a component of oxygen uptake less sensitive to cyanide which was completely inhibitable by low concentrations of decanol . From this it was estimated that in vivo luciferase is responsible for less than 12% (Vibrio harveyi) or 20% (Vibrio fischeri) of the total respiration . From these data in vivo bioluminescent quantum yields are estimated to be not lower than 1.7 and 2.6%, respectively.

J Clin Microbiol, 1986 Feb, 23(2), 373 - 4
Intracranial infection by Vibrio alginolyticus following injury in salt water; Opal SM et al.; A 20-year-old man presented with an epidural abscess 3 months after a seawater diving accident . Cultures of the abscess cavity obtained by surgical drainage revealed a pure culture of Vibrio alginolyticus . Marine vibrios may produce serious intracranial infection after head injury in salt water.

Can J Microbiol, 1986 Feb, 32(2), 99 - 103
Evidence for the presence of a novel biosynthetic pathway for norspermidine in Vibrio; Yamamoto S et al.; Enzymatic studies of the cell extracts of Vibrio alginolyticus and V . parahaemolyticus provided evidence that there exists a novel biosynthetic pathway for norspermidine (NH2(CH2)3NH(CH2)3NH2), a major polyamine species . In this pathway, the Schiff base formed between aspartic beta-semialdehyde and 1,3-diaminopropane is first reduced by a NADPH-dependent enzyme to yield "carboxynorspermidine" (NH2(CH2)3NH(CH2)2CH(NH2)COOH), which is in turn decarboxylated by a pyridoxal phosphate dependent enzyme to form norspermidine . The end product and its intermediate were identified by gas chromatography - mass spectrometry . Experiments with L-{U-14C}aspartic acid resulted in appreciable incorporation of the label into norspermidine . Putrescine could replace 1,3-diaminopropane as a substrate to produce spermidine, but at a reduced rate . The enzyme activity was greatly enhanced by dithiothreitol . Since the activity of an aminopropyltransferase that utilizes decarboxylated S-adenosylmethionine as an aminopropyl group donor could not be detected in any of the cell extracts by our assay method, it was concluded that this novel pathway is primarily responsible for producing norspermidine and spermidine in these species.

J Med Microbiol, 1986 Feb, 21(1), 13 - 7
The effect of lincomycin on exoprotein production by Vibrio cholerae; Young DB et al.; Lincomycin has a differential effect on exoprotein production by Vibrio cholerae . The production of some proteins, such as cholera toxin and deoxyribonuclease, is stimulated by low concentrations of the drug while production of other proteins, such as protease and alkaline phosphatase, is unaffected . Possible mechanisms of the lincomycin effect are discussed.

Acta Med Okayama, 1986 Feb, 40(1), 45 - 53
Augmentation of natural killer activity by neuraminidase treatment of lymphocytes from tumor-bearing mice; Ono M et al.; Spleen cells from tumor-bearing mice showed decreased natural killer (NK) activity and decreased binding to target cells with progression of the tumor . Treatment of spleen cells from tumor-bearing mice with vibrio cholerae neuraminidase (VCN) increased the cytotoxicity to a level twice or more as high as that of untreated cells, but the same treatment of spleen cells from normal mice had no or little effect . On the other hand, neither in spleen cells from tumor-bearing mice nor in those from normal mice, the VCN treatment had no effect on their binding to M-HeLa cells . The suppression of NK activity by preincubation with serum from tumor-bearing mice or prostaglandin E2 was completely abolished by VCN treatment . The above results indicate that VCN treatment of lymphocytes might augment NK activity by an antagonistic effect against an immune suppressive factor.

Am J Chin Med, 1986, 14(1-2), 73 - 83
The effect of needle stimulation of acupuncture loci Tienshu (St-25) Chung-Wan (CV-12) on the immune response in sensitized mice against experimental cholera; Kuan TK et al.; The effects of stimulation of acupuncture loci from Tien-Shu (St-25) piercing through Chung-Wan (CV-12) on the Leukocytes and immune response were assessed in mice (that is, the anatomical equivalent of these loci of human acupuncture points) . The leukocyte count increased and reached its highest level two hours after needling, then restored to normal level 24 hours later both in normal mice and immunized mice . Statistical analysis showed no significant variation on the lymphocyte/total leukocyte ratio in normal mice or immunized mice before, during and after needling . The effects of acupuncture on the production of anti-Vibrio cholerae in serum of the immunized mice can not be found, but produced positive effects in small intestine both on production of SIgA and antagonism to cholera . Furthermore these enhanced effects were higher by needling after oral (p.o.) boosting than that before oral boosting.

Diagn Microbiol Infect Dis, 1986 Jan, 4(1), 49 - 51
Esculin hydrolysis by Vibrio vulnificus; Tison DL; A clinical isolate of Vibrio vulnificus was found to hydrolyze esculin when tested on bile-esculin-azide agar during the initial characterization of the strain . Reports in the literature of esculin hydrolysis by V . vulnificus are conflicting . We tested herein 52 strains of V . vulnificus from clinical and environmental sources for the ability to hydrolyze esculin . Seventy-eight percent of the strains hydrolyzed esculin on bile-esculin-azide agar, whereas all strains of V . vulnificus tested were positive for esculin hydrolysis in a noninhibitory medium, whereas some strains failed to hydrolyze esculin on media containing inhibitory compounds.

Cancer Immunol Immunother, 1986, 23(3), 192 - 9
Tumor therapy of neoplastic diseases with tumor cells and neuraminidase . Further experimental studies on chessboard vaccination in canine mammary tumors; Sedlacek HH et al.; The therapeutic effect of i.d . injection of tumor cells mixed with Vibrio cholerae neuraminidase (VCN) on tumor progression in dogs with spontaneous mammary tumors was investigated . The i.d . injections were performed in a chessboard-like manner: different numbers (10(5), 10(6), 10(7), and 10(8) of mitomycin-treated autologous tumor cells (M-TC) were each mixed with different amounts (10, 50, and 100 mU) of VCN . These different mixtures were injected i.d . at different sites in dogs on the day of resection of a part of multiple tumors.

Dev Comp Immunol, 1986 Summer, 10(3), 341 - 51
Immunization of carp (Cyprinus carpio) with a Vibrio anguillarum bacterin: indications for a common mucosal immune system; Rombout JW et al.; Uptake and transport of formalin-killed Vibrio anguillarum bacteria were studied in the second gut segment of carp and the resulting reaction of the immune system was investigated . Within a few hours after anal administration antigenic determinants of bacteria were present in intraepithelial macrophages of the second gut segment . In gut and skin mucus and bile, immunoglobulins (Ig's) were detected, but the amount was much lower than found in serum . In mucus of the second gut segment, 4 times more Ig (per mg protein) was found than in the first segment . Upon oral or anal immunization, slightly enhanced antigen-specific Ig titers could be detected in skin mucus and bile, but only after a booster along the same route . The existence of a common mucosal immune system is discussed, with special reference to the significance of the second gut segment . After anal intubation an increase of antigen-specific Ig could also be observed in serum . Following a booster, an enhanced cal memory . After anal boosting equal levels of serum antibody were reached compared with two consecutive intramuscular injections . However, no significant antibody increase occurred in serum after oral immunization, not even when bacteria were administered daily with the food.

Dev Comp Immunol, 1986 Summer, 10(3), 295 - 304
Serological characterization of humoral lectin from Heterometrus granulomanus scorpion hemolymph; Ahmed H et al.; A lectin in the hemolymph of Indian scorpion Heterometrus granulomanus was detected by agglutination of human and animal erythrocytes . The agglutinating activity was enhanced in presence of Ca2+ ion . The lectin shows specificity for sialic acid like many other sialic acid-specific lectins such as "Limulin", as the agglutination of erythrocytes was completely abolished by treatment with Vibrio cholerae neuraminidase . However, neuraminidase-treated rat and mouse erythrocytes exhibited high titers . This result was substantiated by crossed-absorption test which suggests the adjunct specificity of the Heterometrus lectin for these cells . Hemagglutination-inhibition results of the lectin indicate that sialic acid including its derivatives and sialoglycoconjugates are the inhibitors among which glycophorin was most potent.

Bull Soc Pathol Exot Filiales, 1986, 79(3), 305 - 12
{Antibiotic resistance of strains of Vibrio cholerae eltor isolated in Douala (Cameroon)}; Garrigue GP et al.; Endemic cholera has been prevalent in Douala since 1972, with sudden epidemic outbreaks occurring every two years during the dry season . The massive and systematic use of chemoprophylaxis since April, 1983 has led to the selection of strains of Vibrio cholerae eltor that are resistant to sulphamide and tetracycline . During the 1984-1985 epidemic, 89.3% of the isolated strains were resistant to sulphamides, 87.5% to a sulfamethoxazole-trimethoprim combination and to the 0/129 disk, 55.3% to tetracycline, 91.1% to chloramphenicol, 73.2% to streptomycin and 94.6% to ampicillin . The epidemic aspect of this multiple resistance to antibiotics raises the issue of the role of a group C incompatibility resistance plasmid . As regards prophylaxis, until hygiene conditions can be improved, which is the only way cholera can be eradicated from our region, vaccination with oral vaccines such as that of the Institut Pasteur seems to be the best way of preventing further epidemics.

J Biochem (Tokyo), 1986 Jan, 99(1), 311 - 4
Occurrence of tetrodotoxin and anhydrotetrodotoxin in Vibrio sp . isolated from the intestines of a xanthid crab, Atergatis floridus; Noguchi T et al.; Vibrio sp . isolated from a xanthid crab, Atergatis floridus, was cultured, and tetrodotoxin (TTX) and anhydroTTX were indicated to be present in several fractions of the cell extract and the culture medium by reverse phase HPLC . The presence of the C9-base in alkaline hydrolyzates of these fractions was confirmed by GC-MS and UV spectrometry . These results showed the production of TTX and anhydroTTX in the Vibrio sp., thus indicating one of the origins of TTX in nature.

Antonie Van Leeuwenhoek, 1986, 52(2), 145 - 52
Survival of Vibrio parahaemolyticus in cold smoked fish; Karunasagar I et al.; Fresh samples of mullet (Mugil cephalus) and oil sardines (Sardinella longiceps) obtained from a fish market were subjected to cold smoking . Some of the samples harboured low levels of Vibrio parahaemolyticus . After cold smoking, however, many samples showed relatively high levels of V . parahaemolyticus suggesting that a small population of naturally occurring organisms could multiply to significant levels during the process of cold smoking or during subsequent storage at room temperature . Nevertheless, smoke components were observed to exert an inhibitory effect on V . parahaemolyticus in broth . Salt concentration 1% appeared to increase the sensitivity of V . parahaemolyticus to smoke components.

Trans R Soc Trop Med Hyg, 1986, 80(1), 60 - 3
Epidemiological differences between cholera due to multiple antibiotic resistant and multiple antibiotic sensitive Vibrio cholerae infection; Khan MU et al.; The appearance of cholera caused by multiply antibiotic resistant Vibrio cholerae in Bangladesh provided an opportunity to compare epidemiological features of infection caused by resistant and by sensitive V . cholerae . A prospective study was carried out using 46 families of hospital in-patient cholera cases due to resistant V . cholerae and 11 families of hospital cases due to sensitive V . cholerae and nine cases of cholera due to resistant and six cases due to sensitive V . cholerae detected in the neighbourhoods of hospital patients . All families were visited daily during ten days for cultures of rectal swabs, samples of domestic water and for history of diarrhoea . The results showed no significant difference in secondary infection and case rates in contacts of hospital cholera cases due to resistant and sensitive V . cholerae . However, the secondary infection rate (57%) in contacts of cases due to resistant V . cholerae detected from the neighbourhoods of hospital cases was significantly higher (p less than 0.05), than in the neighbourhood case-contacts (29%) of cases due to sensitive V . cholerae . The mean duration of diarrhoea in untreated resistant V . cholerae cases who were contacts of hospital cases (3.3 days) was significantly longer (p less than 0.05) than that of untreated sensitive V . cholerae (2.2 days) . Higher isolation rates of V . cholerae were obtained from water sources used by cholera cases due to resistant V . cholerae, than from sources used by cases due to sensitive V . cholerae, but the differences were not statistically significant (p greater than 0.05) . The study suggests that resistant V . cholerae poses an additional threat through a higher secondary infection rate and by causing illnesses of longer duration.

Microbiol Immunol, 1986, 30(3), 193 - 201
Composition of major outer membrane proteins of Vibrio vulnificus isolates: effect of different growth media and iron deficiency; Koga T et al.; The outer membrane proteins of Vibrio vulnificus including isolates from humans, seawater and an asari clam were examined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis . A major outer membrane protein with an apparent molecular weight of 48,000 (48K protein) was common to all the strains grown in 3% NaCl-nutrient broth; however this 48K protein was not produced in any of the strains grown in chemically defined medium . Other major outer membrane proteins with molecular weights ranging from 33,000 to 40,000 varied in number, relative amount and molecular weight depending on the strain . One to three new outer membrane proteins with molecular weights ranging from 74,000 to 85,000 were produced in the cells grown in iron-deficient medium . The 48K protein and one or two major proteins with molecular weights ranging from 35,000 to 37,000 in the cells grown in 3% NaCl-nutrient broth were not solubilized by 2% SDS at 60 C for 30 min and were resistant to trypsin, indicating that they are porins . On the other hand, in cells grown in chemically defined medium, one or two major outer membrane proteins with molecular weights ranging from 33,000 to 40,000 might be porins.

Can J Microbiol, 1986 Jan, 32(1), 71 - 3
Purification of the thermostable direct hemolysin of Vibrio parahaemolyticus by immunoaffinity column chromatography; Honda T et al.; A simple method involving immunoaffinity column chromatography to purify the thermostable direct hemolysin of Vibrio parahaemolyticus was developed . The thermostable direct hemolysin purified from the culture supernatant of a strain isolated from the first reported case of V . parahaemolyticus infection in China in 1985 was indistinguishable from the hemolysins purified from strains isolated in Japan.

Rev Epidemiol Sante Publique, 1986, 34(6), 419 - 26
{An epidemic of cholera in the city of Pemba (Mozambique) in 1983}; Cans C; The purpose of the epidemiological study about 205 suspected hospitalized cases of cholera was to determinate which model of transmission was the most important . From the results, we noticed that direct non-waterborne has been more frequent than waterborne transmission (mainly in the infection of family contacts of index cases) . We also found that virulence of the Vibrio cholerae El Tor responsible was quite close of virulence of the Vibrio cholerae classical.

Microbiol Immunol, 1986, 30(11), 1075 - 83
Culture conditions for stimulating cholera toxin production by Vibrio cholerae O1 El Tor; Iwanaga M et al.; A method that stimulates cholera toxin (CT) production by Vibrio cholerae O1 biotype El Tor (El Tor vibrios) to the level of several micrograms per ml in the culture fluid was established . Such a large amount of CT was obtained by the following method: El Tor vibrios were cultured in AKI medium (1.5% Bacto peptone, 0.4% yeast extract-Difco, 0.5% NaCl, 0.3% NaHCO3) at 37 C for 4 hr in a stationary test tube and then for 16 hr in a shaken flask, with inoculum sizes of 10(5) to 10(7)/ml . With this method, 35 strains out of 60 examined produced 2 to 16 micrograms/ml of CT as determined by the reversed passive latex agglutination test (RPLA) . Thirty-three randomly selected strains out of the 60 produced reasonable amounts of rabbit skin vascular permeability factor, reflecting the amount of CT titrated with RPLA.

Microbiol Immunol, 1986, 30(5), 437 - 44
Mechanisms of plasmid-mediated antibiotic resistances in Vibrio parahaemolyticus; Hasegawa H et al.; The mechanisms of drug resistance of clinical isolate, Vibrio (V.) parahaemolyticus ST550, resistant to chloramphenicol (CP), aminoglycoside antibiotics (AGs) and beta-lactam antibiotics were investigated . The mechanisms of resistance to CP, AGs and beta-lactam antibiotics were dependent on chloramphenicol acetyltransferase (CAT), aminoglycoside-3"-adenylyltransferase AAD(3") and aminoglycoside-3'-phosphotransferase APH(3') and TEM type penicillinase, respectively.

Arch Microbiol, 1986 Jan, 143(4), 325 - 9
Mobilization of cloned luciferase genes into Vibrio harveyi luminescence mutants; Gupta SC et al.; A recombinant plasmid which carried a 5 kb fragment of Vibrio harveyi DNA containing the luxA and luxB genes was mobilized from Escherichia coli into luminescence-deficient mutants of V . harveyi . The cloned genes complemented a temperature sensitive luciferase mutation, but failed to complement lesions in two different aldehyde deficient mutants . Expression of the cloned genes was not subject to autoinduction in either E . coli or in V . harveyi.

Hokkaido Igaku Zasshi, 1986 Jan, 61(1), 24 - 34
{Enterotoxin-like factor(s) produced by Vibrio parahaemolyticus}; Kimura K; Culture filtrates obtained from several isolates of Vibrio parahaemolyticus were tested for their enterotoxin-like activities . Also their antigenic relationship with cholera enterotoxin and Kanagawa hemolysin were tested . In the Chinese Hamster Ovary (CHO) cells assay which are usually employed for detecting cholera toxin or heat labile enterotoxin produced by enterotoxigenic Escherichia coli, crude culture filtrates obtained from both Kanagawa positive and negative strains isolated from patients showed positive reaction whereas most of the Kanagawa negative strains isolated from environmental sources did not . After concentration, however, both the culture filtrates of Kanagawa positive and negative strains of Vibrio parahaemolyticus showed positive reactions in CHO cell assay and rabbit skin permeability assay but not in rabbit ileal loop test . Antisera against concentrated culture filtrates obtained from both Kanagawa positive and negative strains neutralized neutralized the activities of concentrated culture filtrates in CHO cell assay and Rabbit permeability assay but not of cholera toxin and Kanagawa hemolysin . Immunodiffusion test was also carried out using the antisera and the concentrated culture filtrates obtained from both of Kanagawa positive and negative strains of Vibrio parahaemolyticus . The antisera cross-reacted with all concentrated culture filtrates obtained from both Kanagawa positive and negative strains but not reacted with cholera toxin and Kanagawa hemolysin . These results indicated that both of Kanagawa positive and negative strains of Vibrio parahaemolyticus can produce the enterotoxin-like factor(s) such as cholera enterotoxin or heat labile enterotoxin produced by enterotoxigenic Escherichia coli, but this factor(s) dose not show cross-antigenicity with cholera enterotoxin and the Kanagawa hemolysin produced by Kanagawa positive Vibrio parahaemolyticus.

Gene, 1986, 45(2), 203 - 9
Luciferase genes cloned from the unculturable luminous bacteroid symbiont of the Caribbean flashlight fish, Kryptophanaron alfredi; Haygood MG et al.; Light organs of anomalopid (flashlight) fish contain luminous bacteroids that have never been cultured and, consequently, have been difficult to study . We have characterized the luciferase (lux) region of DNA extracted from light organs of the Caribbean flashlight fish Kryptophanaron alfredi by hybridization of cloned Vibrio harveyi lux genes to restriction-endonuclease-digested, light organ DNA . Comparison of the hybridization pattern of light organ DNA with that of DNA of a putative symbiotic isolate provides a method for identifying the authentic luminous symbiont regardless of its luminescence, and was used to reject one such isolate . Light organ DNA was further used to construct a cosmid clone bank and the luciferase genes were isolated . Unlike other bacterial luciferase genes, the genes were not expressed in Escherichia coli . When placed under the control of the E . coli trp promoter, the genes were transcribed but no luciferase was detected, suggesting a posttranscriptional block to expression.

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