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) caus