J Infect Dis, 1982 Mar, 145(3), 296 - 9 The pathogenicity of nonenterotoxigenic Vibrio cholerae serogroup O1 biotype El Tor isolated from sewage water in Brazil; Levine MM et al.; Nonenterotoxigenic strains no . 1196-78 and no . 1074-78 of Vibrio cholerae serogroup O1 (biotype El Tor, serotype Ogawa) were isolated from sewage water in Brazil and fed to 20 volunteers . Neither strain caused diarrhea . None of the seven volunteers who ingested Ogawa strain no . 1074-78 (10(6) organisms) excreted the organism whereas eight of the 13 volunteers who ingested Ogawa strain no . 1196-78 (10(6) or 10(8) organisms) did excrete the organism in their stools . None of 114 stool-culture isolates yielded cholera enterotoxin, and none of the 20 volunteers had significant increases in serum titers of antitoxin as measured by enzyme-linked immunosorbent assay although six of the volunteers had slightly elevated vibriocidal antibody levels . Six volunteers used as controls and four volunteers who had stool cultures positive for strain no . 1196-78 of V . cholerae were challenged with pathogenic El Tor Ogawa strain no . E7946 (10(6) organisms) to determine if previous ingestion of the Brazilian strain would induce protective immunity . All 10 of the volunteers developed diarrhea, and the severity of the illness was similar in both groups. Eur J Biochem, 1982 Mar 1, 122(3), 581 - 6 Role of sialic acid in the mobility of membrane proteins containing O-linked oligosaccharides on polyacrylamide gel electrophoresis in sodium dodecyl sulfate; Gahmberg CG et al.; The major sialoglycoprotein of the human red-cell membrane, glycophorin A, contains 15 O-glycosidically linked oligosaccharides and one N-glycosidic oligosaccharide . The protein shows a decreased mobility on polyacrylamide gel electrophoresis in sodium dodecyl sulfate after neuraminidase treatment of the non-denatured protein . The molecular mechanism behind this phenomenon has been elucidated . Neuraminidase treatment of glycophorin A in intact cells or after solubilization in buffers containing Triton X-100 resulted in conversion of the predominant tetrasaccharide N-acetylneuraminosyl alpha 2-3galactosyl beta 1-3(N-acetylneuraminosyl alpha 2-6)-N-acetylgalactosamine to the trisaccharide galactosyl beta 1-3(N-acetylneuraminosyl alpha 2-6)-N-acetylgalactosamine and the disaccharide galactosyl beta 1-3-N-acetylgalactosamine . After denaturation with sodium dodecyl sulfate, Vibrio cholerae neuraminidase also liberated the N-acetylgalactosamine-bound sialic acids . Such treatment resulted in increased electrophoretic mobility . The results show that distal sialic acids linked to galactose are readily available to neuraminidase, and that their negative charge gives an increased electrophoretic mobility in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate . In contrast, most of the N-acetylgalactosamine-linked sialic acids of glycophorin A are not liberated by neuraminidase without denaturation of the substrate . Like sialic acids of complex-type oligosaccharides the decreased electrophoretic mobility caused by them is exclusively due to their mass while no significant contribution by the charge was seen. Afr J Med Med Sci, 1982 Mar, 11(1), 11 - 8 Characteristics of boric acid tolerant Vibrio cholerae; Ojo MO; Two strains of boric acid tolerant Vibrio cholerae have been studied in order to identify the characteristics which distinguish them from the 'wild-type' virulent strains . Strain Ib5 requires either amino acid cysteine or methionine while strain Ib5S requires deoxyribonucleic acid (DNA) for growth . Although the strains did not grow on desoxycholate citrate agar (DCA) they grew well on chocolate agar . Morphologically filamentous, bacillary and comma-shaped forms were seen, depending on the media and incubation period . The strains failed to cause accumulation of fluid in gut ligation and permeability factor tests . With the single radial immunodiffusion test, strain Ib5S gave positive reaction in 83% of the tests when DNA was added to the growth medium . On the other hand only 25% of the tests were positive when no DNA was added to the growth medium, even though the antigen was concentrated ten-fold . Strain Ib5 only gave 50% positive tests when the antigen was concentrated ten-fold . It is therefore suggested that Ib5S strain is a cholerangenoid producer in the presence of DNA, and may be useful as an effective vaccine against cholera. Infect Immun, 1982 Mar, 35(3), 1155 - 6 Collagenolytic activity of Vibrio vulnificus: potential contribution to its invasiveness; Smith GC et al.; Vibrio vulnificus (lactose-positive vibrio) produced collagenase when grown in 2% synthetic sea salts supplemented with hydrolyzed casein . The addition of collagen or peptone to the medium increased the level of collagenase production . Collagenase activity was inhibited by EDTA but not by fetal calf serum. Biochim Biophys Acta, 1982 Feb 10, 720(1), 65 - 74 Influenza virus: an NMR study of mechanisms involved in infection; Mountford CE et al.; High resolution 1H-NMR spectroscopy has been used to study the infection of chicken embryo fibroblasts by influenza virus . Marked changes in the NMR spectrum occur when infectious influenza virus is introduced into the fibroblasts and these changes appear to depend upon the presence of active neuraminidase (EC 3.2.1.18) . A crude preparation of neuraminidase from Vibrio cholerae is able to effect similar changes . Only minor spectral changes are observed in the absence of culture medium or when the viral genome material is inactivated by beta-propiolactone . Similarly, little change is seen in the NMR spectrum when amantadine, which is thought to inhibit uncoating of the virus inside the cell, or actinomycin D, which inhibits cellular nucleic acid metabolism, are incubated with fibroblasts prior to the addition of virus . The results suggest that neuraminidase, in co-operation with a factor in the infectious process, initiates a cellular event which can be monitored by NMR . The nature of this cellular mechanism is unknown, but further studies are under way to determine its importance in viral infection. J Med Microbiol, 1982 Feb, 15(1), 53 - 61 Role of somatic antigen of Vibrio cholerae in adhesion to intestinal mucosa; Chitnis DS et al.; The in-vitro adhesion of Vibrio cholerae to intestinal mucous membrane was studied in isolated adult-rabbit ileal loops . Antisomatic antiserum Against V . cholerae Inaba could inhibit adhesion of three different strains of V . cholerae Inaba but had no effect on the adhesion of two different strains of enterotoxigenic NAG vibrios . The antiserum's bacterial agglutinin titre was 320, its anti-Inaba lipopolysaccharide (LPS) titre was 16 000 and its anti-flagellar antibody titre was 3200 . Conversely, anti-live V . Cholerae Inaba antiserum absorbed with boiled cells of Inaba and devoid of antisomatic antibody, could not inhibit adhesion of the same three strains of V . cholerae Inaba . This antiserum had no anti-LPS or bacterial agglutinin activity, but its anti-flagellar antibody titre was 32 000 . Thus, ability to inhibit adhesion of V . cholerae could be correlated only with antisomatic (anti-LPS) antibody activity . Antisomatic antiserum had no activity against 'adhesion', a V . cholerae surface antigen described by Freter . Conversely anti-live V . cholerae antiserum absorbed with boiled cells showed anti-adhesion activity even at a dilution of 1 in 200 . LPS preparations from V . cholerae strain 569B Inaba could inhibit adhesion of two different Inaba strains to the intestinal mucous membrane . It is concluded that the somatic antigen plays a major role in the adhesion of V . cholerae to the intestinal mucous membrane. J Med Microbiol, 1982 Feb, 15(1), 43 - 51 Role of bacterial adhesion in the pathogenesis of cholera; Chitnis DS et al.; In studies with the adult-rabbit ileal-loop model, antibodies to the lipopolysaccharide somatic-antigen component of Vibrio cholerae gave passive protection against challenge with live V . cholerae . The antisomatic antibodies had no effect on bacterial proliferation and toxin production either in vivo or in vitro; after challenge, antibody-protected and non-protected rabbit ileal loops developed almost identical amounts of cholera toxin and numbers of V . Cholerae . The protection could be correlated only with a 10-15-fold reduction in the number of V . cholerae adherent to the mucous membrane of the antibody-protected loops . The amount of cholera toxin in the two sets of loops ranged from 1600 to 3200 units . In contrast, when biologically active cholera toxin was prepared in vitro, the amount required to induce ileal-loop secretion was very large (25,600 units) . These findings indicate that toxin production by adherent vibrios on the surface of the mucous membrane is an important factor in the pathogenesis of cholera. Zh Mikrobiol Epidemiol Immunobiol, 1982 Feb, (2), 63 - 70 {Quantitative evaluation of Vibrio cholerae colonization and lymphocyte count in small intestine villi of rabbits immunized with vibrio and cholera toxin}; Efremov VE et al.; The influence of parenteral immunization with live virulent vibrios and lipopolysaccharide-containing choleragen toxoid and oral immunization with liver avirulent vibrios on the interaction of V . eltor and the mucous membrane was studied on the ligated loops of the small intestine in 164 rabbits . Protection from enterotoxigenicity (the accumulation of fluid) was compared with the suppressive effect on the adhesion and colonization of vibrios . The coefficient of immunization efficacy was determined microbiologically, as the ratio of the adhesion index (i . e . the percentage of the number of vibrios in the media inoculated with the homogenized matter of the washed intestinal wall from the sum of this number and the amount of vibrios in the contents) in the control rabbits to the adhesion index in the immunized ones, and histologically, as the ratio of the average number of vibrios attached to the epithelial number of vibrios attached to the epithelial cells of one intestinal villus in the control rabbits to the corresponding number in the immunized animals . Protection from the enterotoxigenicity of vibrios resulted from the suppression of their adhesion and colonization, lipopolysaccharide being, probably, the factor of adhesiveness . Effective protection was accompanied by an increase in the number of lymphocytes (especially "granular" lymphocytes with lysosome-like granules) between intestinal villi and the number of lymphoid and plasmic cells in the lamina propria of the villi, which indicates the role of lymphocytes in cholera-induced immunity. J Gen Microbiol, 1982 Feb, 128(Pt 2), 349 - 53 Repression of the alkaline phosphatase of Vibrio cholerae; Roy NK et al.; The synthesis of alkaline phosphatase by two strains of Vibrio cholerae belonging to the Inaba and Ogawa serotypes has been examined in relation to the phosphate concentration of the culture medium . The synthesis of the enzyme in both strains was repressed in cells grown in the presence of a high concentration of inorganic phosphate . Lowering the phosphate content of the growth medium led to a derepression of enzyme activity . The presence of glucose in low phosphate medium stimulated the degree of derepression . The synthesis of the enzyme by strain Inaba 569B was more sensitive to inorganic phosphate than that of strain Ogawa 154 . The enzyme was presumably located in the periplasmic space since it was released when the organisms were converted to spheroplasts. Antibiotiki, 1982 Feb, 27(2), 126 - 9 {NAG vibrio sensitivity to antibiotics}; Andrusenko IT et al.; Sensitivity of 575 NAG-vibrio strains isolated from various substrates in 1978-1980 to 14 antibiotics was studied and compared with the results of an analogous study performed in 1968 by Z . V . Ermolyeva et al . It was shown that sensitivity of NAG-vibrio to chloramphenicol, ampicillin, cephaloridine and streptomycin decreased . However, NAG-vibrio remain to be sensitive to tetracycline, chloramphenicol and gentamicin . Resistance of the majority of the strains to ampicillin, carbenicillin, cephaloridine and polymyxin M was noted. J Infect Dis, 1982 Feb, 145(2), 248 - 54 Cholera-like illness due to Aeromonas sobria; Champsaur H et al.; A Thai woman from Bangkok was admitted to a hospital in Paris for a cholera-like illness . A culture of her "rice-water" diarrhea was negative for Vibrio cholerae and enterotoxigenic Escherichia coli but was positive for Aeromonas sobria . This strain produced enterotoxin, cytolysin, proteolysin, hemolysin, and a cell-rounding factor . Acute -and convalescent-phase sera showed an increase in neutralizing antibodies to enterotoxin, cytolysin, and hemolysin . The enterotoxin, which was labile at 100 C, induced an accumulation of fluid in the rabbit ileal loop model and was not neutralized by antiserum to cholera toxin . Suckling mouse assays and rabbit permeability skin tests were negative, and the Y1 mouse adrenal cell assay produced not true cytotonic effect . This report, the first of an infection due to A . sobria alone, provides evidence that A . sobria is an enteric pathogen of humans that can cause a toxin-mediated, life-threatening illness. Genetika, 1982 Feb, 18(2), 227 - 34 {Mobilization of Vibrio cholerae chromosomal genes by plasmid RP4::Mu cts62}; Skavronskaia AG et al.; The RP4::Mu cts62 plasmid was constructed in Escherichia coli cells and subsequently transferred by conjugation into the cells of Vibrio cholerae . This plasmid had been shown to mobilize chromosomal genes of V . cholerae during intrageneric matings . The frequency of mobilization is higher in matings at 37 degrees C, the temperature which is semipermissive for temperature sensitive Mu cts62 phage . This is only true for strains harbouring RP4::Mu cts62, but not for strains containing the RP4 plasmid . Frequencies of mobilization per transferred plasmid exceeded conspicuously the known frequency for the mobilizing activity of the P-factor of V . cholerae . Transconjugants selected for chromosomal markers carried no traces of RP4 plasmid or Mu cts62 markers . This feature of their use in the genetical analysis of V . cholerae . Reasons for the absence of plasmid markers in recombinants are being discussed. Infect Immun, 1982 Feb, 35(2), 702 - 8 Detection of toxins produced by vibrio fluvialis; Lockwood DE et al.; The results of studies with cell-free extracts and culture supernatant fluids of Vibrio fluvialis (a recently recognized, potential enteric pathogen for humans) grown in the absence and presence of lincomycin indicated that the bacterium could produce (i) a factor which causes CHO cell elongation (CEF) similar to that elicited by V . cholerae enterotoxin and by the heat-labile enterotoxin of Escherichia coli, (ii) cytolysin(s) active against erythrocytes, (iii) nonhemolytic, CHO cell-killing factor(s), and (iv) protease(s) active against azocasein . The CEF was heat labile and ammonium sulfate precipitable, and it had an isoelectric point (estimated by sucrose density gradient electrofocusing) and molecular weight (estimated by gel filtration) of about 5.1 and 135,000, respectively. Infect Immun, 1982 Feb, 35(2), 471 - 5 Berberine inhibits intestinal secretory response of Vibrio cholerae and Escherichia coli enterotoxins; Sack RB et al.; Berberine, an alkaloid from the plant Berberis aristata, which has been known since ancient times as an antidiarrheal medication in India and China, inhibited by approximately 70% the secretory responses of the heat-labile enterotoxins of Vibrio cholerae and Escherichia coli in the rabbit ligated intestinal loop model . The drug was effective when given either before or after enterotoxin binding and when given either intraluminally or parenterally; it did not inhibit the stimulation of adenylate cyclase by cholera enterotoxin and caused no histological damage to intestinal mucosa . Berberine also markedly inhibited the secretory response of E . coli heat-stable enterotoxin in the infant mouse model . Although the mechanism of action of the drug is not yet known, these data provide a rationale for its apparent clinical usefulness in treating acute diarrheal disease. Infect Immun, 1982 Feb, 35(2), 391 - 5 Potentiating effect of bile on enterotoxin-induced diarrhea; Lange S et al.; The influence of bile acids on adenosine 3',5'-phosphate-induced intestinal secretion was studied in mice . Bile flow was stopped by ligation of the common bile duct, and secretion was induced in ligated loops of the small intestine . The decrease of bile led to inhibition of hypersecretion after challenge with heat-labile enterotoxins from Vibrio cholerae and Escherichia coli, as well as with prostaglandin E1 . In contrast, the fluid response induced by dibutyryl-adenosine 3',5'-phosphate was unaffected by intestinal bile . Injection of bile or bile acids into intestinal loops before cholera toxin challenge restored the toxin-induced secretion in the bile-depleted intestine . At the subcellular level the decrease of intestinal bile led to inhibition of cholera toxin-activated adenylate cyclase, whereas the bile concentration did not influence the binding of 125I-labeled toxin to the intestinal epithelial cells . The results suggest that intestinal bile interacts with adenylate cyclase in the induction of fluid secretion by enterotoxins and prostaglandin E1. J Biol Chem, 1982 Jan 25, 257(2), 788 - 94 Roles of Na+ and K+ in alpha-aminoisobutyric acid transport by the marine bacterium Vibrio alginolyticus; Tokuda H et al.; Effects of monovalent cations on alpha-aminoisobutyric acid (AIB) transport were examined in the marine bacterium Vibrio alginolyticus . In K+-containing cells, AIB was actively accumulated only in the presence of Na+, and the addition of K+ had essentially no effect . On the other hand, K+-depleted and Na+-loaded cells required K+ as well as Na+ for the accumulation of AIB against its concentration gradient . The characterization of the roles of Na+ and K+ in AIB transport was performed by manipulation of intra- and extracellular cation compositions . K+ concentration gradient (K+in greater than K+out) was not essential for the Na+-dependent AIB uptake . Na+ extrusion against its concentration gradient in Na+-loaded cells occurred only in the presence of K+(Rb+) . Half-maximal stimulations of the Na+ extrusion and AIB uptake by K+ were observed at K+ concentration near apparent Km for K+ transport . Finally, in the presence of the Na+ electrochemical gradient (toward the inside), K+ was not necessary for AIB uptake . From these results, it was concluded that the Na+-dependent AIB uptake is driven by the Na+ electrochemical gradient across the membrane and that K+ is required for AIB uptake only for the generation of the Na+ electrochemical gradient. G Batteriol Virol Immunol, 1982 Jan-Jun, 75(1-6), 128 - 34 {Isolation and significance of Vibrio cholera NAG}; Piantieri G et al.; After the isolation of two Vibrio cholerae NAG from the stools of two tourists, the authors researched Vibrio in people coming home from particular countries and in resident people . The research was extended to the water of Varese lake after another isolation from a fisher who had fished, cooked and eaten the lake fish . Problems concerning the classification of Vibrio and their presence in the environment are examined. Trans R Soc Trop Med Hyg, 1982, 76(5), 590 - 4 Endotoxaemia in Vibrio El Tor cholera; Onyemelukwe GC et al.; Endotoxaemia and endotoxin-induced changes were sought in Nigerian patients presenting with cholera/diarrhoea . The organism was Vibrio cholerae, bio-type El Tor, serotype Hikojima . The limulus amoebocyte lysate gelation test was used qualitatively by the clot method, whilst a spectrophotometric method was used quantitatively to measure endotoxin levels . 25 acutely ill patients tested had detectable endotoxaemia by the Escherichia coli endotoxin standard . The highest endotoxin level was found in a patient with sub-conjunctival haemorrhage . Changes in platelet counts, the detection of complement breakdown product C3d in plasma, the elevation of fibrin degradation products, the finding of elevated, normal or depressed C3 levels and the absence of circulating immune complexes, suggest a pathogenic role for endotoxin in Vibrio cholerae El Tor diarrhoea. Antibiotiki, 1982, 27(12), 18 - 22 {Determination of the sensitivity of NAG-Vibrio to antibiotics by the methods of serial dilution in agar and agar diffusion}; Givental' NI et al.; The methods of serial dilutions and agar diffusion in medium AGV were used in the study on sensitivity of 200 NAG-vibrio strains to 10 antibiotics . The regression equations showing the relation between the MIC log2 and the diameter of the zones of the vibrio growth inhibition around the standard discs with the antibiotics were estimated . An evaluation table was developed for semiquantitative interpretation of the diameters of the inhibition zones of the vibrio growth when the agar diffusion method was used . A possibility of rapid estimation of the results in determination of the vibrio sensitivity to antibiotics with the methods of serial dilutions and agar diffusion is shown. Trans R Soc Trop Med Hyg, 1982, 76(6), 786 - 9 Kanagawa phenomenon and serotypic pattern of Vibrio parahaemolyticus strains isolated from various sources in Calcutta; Saha MR et al.; A total of 135 strains of Vibrio parahaemolyticus isolated during the period 1976-77 from human gastroenteritis cases and various categories of environmental sources, e.g., crustaceans, fish and different water samples (pond, stored, open well and tap water) were tested for their Kanagawa phenomenon and serotypic pattern . Amongst the 80 human isolates tested, 69 strains (86.3%) were Kanagawa phenomenon positive, and 60 strains (75.0%) were serologically typable--the dominant serotype being 05:K15 . Of the 25 isolates from various water sources 10(40.0%) were Kanagawa phenomenon positive, and 17(68.0%) were serologically typable and 4 (23.5%) of them belonged to serotype 05:K15 . All the 30 strains isolated from crustaceans and fish were Kanagawa phenomenon negative and 22 (73.3%) of them were serologically typable, belonging to heterogeneous serotypes . The results of this study also indicate the possible role of water in the transmission of V . parahaemolyticus infection in Calcutta slums. Mikrobiologiia, 1982 Jan-Feb, 51(1), 114 - 7 {Micavibrio admirandus gen . et sp . nov.}; Lambina VA et al.; Micavibrio gen . nov . (M.L . fem . n . mica--a tiny thing and M.L . masc . n . Vibroia generic name, M.L . masc . n . Micavibrio a tiny vibrio) . Cells are single, small curved rods, in motile stage with a single polar not sheathed flagellum . Reproduced by binary division, gram-negative . Cells attach to host-bacteria cells and grow as exoparasites destroying the host, may exhibit a host-range specificity . The G + C content of the DNA of the strain examined is 57,1% M . Isolated from sewage waters . The type species is M . admirandus sp . nov . M . admirandus sp . nov (M.L . adj . admirable) . Curved gram-negative rods 0,25-0,35X0,6-1,0 micron with a single flagellum 13-15 nm in width . Motile cells attach to the cell wall of the bacterium . Pseudomonas maltophilia, lose the motility and grow as exoparasites destroying the host . Develops only on . P . maltophilia cells and is unable to multiply in the absence of a host organism . Resistant to a vibriostatic agent pteridine 0/129 . The G + C content of the DNA of the strain examined is 57,1% M . The type strain ARL-14 is maintained in the culture collection of the Institute of Biochemistry and Physiology of Microorganisms, USSR Academy of Sciences . The type strain has been isolated from sewage waters of the town of Pushchino. Comp Biochem Physiol B, 1982, 71(3), 545 - 8 Failure of in vivo and in vitro resialosylation of VCN-desialylated erythrocytes in mammalian and non-mammalian species: evidence from agglutination studies with peanut-agglutinin; Perret G et al.; 1 . In distinction to mammalian erythrocytes, splitting of sialic acids of non-mammalian erythrocytes (chicken and newt) by Vibrio cholerae neuraminidase (VCN) does not lead to a dramatic decrease in their viability . 2 . One of the possibilities to explain this discrepancy is the ability of chicken and newt desialosylated erythrocytes to repair the enzymatic injury . 3 . To test this hypothesis we used the property of peanut agglutinin (PNA) to agglutinate desialosylated erythrocytes . 4 . Desialosylated erythrocytes of all the species tested remain PNA-agglutinable throughout their whole life time in the blood stream . The absence of effect of VCN on chicken and newt erythrocytes is therefore not due to their resialosylation. Can J Microbiol, 1982 Jan, 28(1), 111 - 6 Seasonal distribution of bdellovibrios at the mouth of the Patuxent River in the Chesapeake Bay; Williams HN et al.; Water samples taken at monthly intervals from three sites in the mouth of the Patuxent River in the Chesapeake Bay were cultured for bdellovibrios lytic to Vibrio parahaemolyticus and for total viable bacterial counts . The number of bdellovibrios recovered decreased from the spring months (April, May, June (AMJ) until very few were detected during the winter months (January, February, March (JFM), which also coincided with the lowest water temperatures . During the AMJ season there was a significant increase as compared with the JFM season in the number of bdellovibrios for all sites . The highest number of bdellovibrios was recovered during each season from the shoreline water sample, with one exception . The seasonal variation in the number of bdellovibrios was observed to correlate statistically with the water temperature. Am J Trop Med Hyg, 1982 Jan, 31(1), 128 - 30 Vibrio parahaemolyticus: a major cause of travelers' diarrhea in Bangkok; Sriratanaban A et al.; Travelers' diarrhea in Bangkok, Thailand was investigated in a group of 146 hotel guests who had diarrhea during their stay in the city . The identified pathogenic organisms were Vibrio parahaemolyticus in 45 (31%), Vibrio cholerae in 1, nonagglutinable Vibrio in 1, and Entamoeba histolytica in 3 . Identification of enterotoxigenic Escherichia coli was not attempted . The study suggests that Vibrio parahaemolyticus is a major cause of travelers' diarrhea in Bangkok. Proc Natl Acad Sci U S A, 1982 Jan, 79(1), 151 - 5 Isolation of enterotoxin structural gene deletion mutations in Vibrio cholerae induced by two mutagenic vibriophages; Mekalanos JJ et al.; Phenotypically nontoxinogenic mutants of Vibrio cholerae were isolated after infection with either of two mutagenic vibriophages, VcAI and VcA2ctsl . DNA isolated from these mutants was analyzed for toxin gene sequences by the Southern blotting method with 32P-labeled probes derived from the cloned A and B subunit genes for the heat-labile enterotoxin of Escherichia coli, designated LT . Several of the mutant isolates were shown by this method to have lost all sequences hybridizing to the LT probes, indicating that these clones contain deletion mutations that removed the structural gene(s) for cholera toxin . The mutants were prototrophic and grew normally, in vitro, demonstrating that the toxin is not essential for the growth and viability of V . cholerae . Moreover, the toxin gene deletion mutants multiplied well in vivo in ligated rabbit intestine . Because of these growth properties and the stability of deletion mutations, these strains are promising candidates for testing as live oral vaccine strains for protection against cholera. Trans R Soc Trop Med Hyg, 1982, 76(4), 497 - 9 The distribution of Vibrio parahaemolyticus serotypes in Kenyan seafish, shellfish, marine water and sediment; Binta MG et al.; Vibrio parahaemolyticus was recovered from 74 of 912 marine samples screened for the organism . Of 74 isolates of V . parahaemolyticus obtained from marine fish, crustacean shellfish (prawns, lobster, crabs), and molluscan shellfish (oysters), and from water and sediment collected off the coast of Kenya, only 33 were positively identified . The isolates were only from seafish and shellfish . The main serotypes were 0,3:K37; 0,3:K40; 0,8:39; 0,10:23; 0,10:K52; and 0,11:K40 . All the serotypes were Kanagawa-negative . The rest of the samples, mainly marine sediment and water, revealed what was described as untypable Vibrios . This is the first report of the organism in this part of the world where no clinical disease is reported. Microbiol Immunol, 1982, 26(6), 479 - 85 Conjugation of drug-resistance plasmids from vibrio anguillarum to Vibrio parahaemolyticus; Hayashi F et al.; Drug-resistant strains of Vibrio anguillarum, a fish pathogen, were isolated from diseased fish in culture ponds . In investigations of these strains, the transfer of resistance to chloramphenicol, tetracycline, and sulfanilamide from multiple resistant organisms to laboratory recipients was observed . The plasmid from V . anguillarum was stably maintained in both recipient strains of Vibrio parahaemolyticus and Escherichia coli . The plasmids isolated from Vibrio anguillarum belong to incompatibility group C . The molecular weight of these plasmids determined by electron microscopic observation was about 103 to 113 megadaltons. Antibiotiki, 1982 Jan, 27(1), 32 - 7 {Antibiotic sensitivity and other properties of bacteria of the genus Aeromonas}; Andrusenko IT et al.; Antibiotic sensitivity and other biological properties of Aeromonas and NAD-vibrios isolated from water of open waterbodies were studied comparatively . It was found that the organisms of these genera had a definite similarity with respect to antibiotic sensitivity, morphological, cultural and some antigenic properties . The differences in the antibiotic sensitivity of these organisms were not sufficient for definition of the taxonomic features. J Clin Immunol, 1982 Jan, 2(1), 20 - 9 Characterization of human peripheral blood T lymphocytes bearing receptors for autologous erythrocytes and T lymphocytes lacking these receptors; Sakane T et al.; When human T cells were treated with neuraminidase from Vibrio cholerae, the capacity of T cells to form rosettes with autologous erythrocytes was markedly enhanced . The neuraminidase-treated T cells wee separated with autologous erythrocytes into autorosetting and nonrosetting cell populations, and these two populations examined for their reactivity to mitogens and B cells and for their regulatory activities . Autorosetting T cells proliferated poorly in response to mitogens and autologous and allogeneic B cells; these cells were particularly enriched for cells capable of becoming concanavalin A-induced suppressor cells . Nonrosetting T cells capable of most actively proliferating in response to the mitogens and the B cells failed to exhibit such suppressor function after concanavalin A activation . Coculture experiments between autorosetting and nonrosetting cells further demonstrated that the nonrosetting T cells were able to potentiate the proliferation of the autorosetting T cells with concomitant expression of the suppressor properties. Acta Microbiol Pol, 1982, 31(3-4), 271 - 8 Studies on alkaline phosphatase, catalase and z-galactosidase in Vibrio el tor under normal and rifampicin resistant conditions; Sinha AM et al.; Alkaline phosphate, catalase and beta-galactosidase activities of Vibrio et tor were decreased after acquisition of resistance towards rifampicin . Zn2+, Mn2+ and EDTA inhibited alkaline phosphatase which is most active with p-nitrophenylphosphate as substrate while Mg2+ was found to suppress alkaline phosphatase activity . Removal of EDTA however, restores the original activity . Rifampicin could not induce mutation of lactose nonfermenting Vibrio el for cells allowing them to grow on lactose as sole carbon source, z-galactosidase which is a constitutive enzyme in this case is repressed by glucose . This repression is overcome by cAMP. Biull Eksp Biol Med, 1982 Jan, 93(1), 62 - 4 {Inhibitory effect of combined administration of neuraminidase and interferon in mice with Rauscher leukemia}; Kobrinskii GD et al.; A study was made of the effect of cholera vibrio neuraminidase and mouse interferon on the development of Rauscher's leukemia in BALB/c mice . It was shown that administration of 1500 units interferon to mice 4 hours before and 5 days after inoculation of Rauscher's leukemia virus prolonged the animals' lifespan . Preliminary treatment of the virus-containing material with neuraminidase prevented the death of one third of the experimental animals . Meanwhile the remainder of the animals inoculated with the same virus showed a longer lifespan . Combined use of both the preparations proved the most effective, enabling the prevention from death of 90% of the experimental animals . The mechanism of antiviral action of neuraminidase and interferon is discussed. J Bacteriol, 1982 Jan, 149(1), 368 - 71 NAD metabolism in Vibrio cholerae; Foster JW et al.; Extracts of Vibrio cholerae were assayed for various enzymatic activities associated with pyridine nucleotide cycle metabolism . The activities measured include NAD glycohydrolase, nicotinamide deamidase, nicotinamide mononucleotide deamidase, and nicotinic acid phosphoribosyltransferase . The results obtained demonstrate the existence in V . cholerae of the five-membered pyridine nucleotide cycle and the potential for a four-membered pyridine nucleotide cycle . The data presented also suggest that most of the NAD glycohydrolase in V . cholerae extracts is not directly related to cholera toxin. Southeast Asian J Trop Med Public Health, 1981 Dec, 12(4), 506 - 12 Protection against Vibrio cholerae infection afforded by fragments of anti-haemagglutinin; Foo Eng Sim A et al.; The finding that Fab fragments of the anti-E1 Tor haemagglutinin were able to afford protection in vivo (low but significant), as well as a significant reduction in Vibrio cholerae adherence to isolated intestinal epithelial cells in-vitro, implicate that masking of these cell-bound haemagglutinin sites per se, would be sufficient to confer protection in E1 Tor cholera infection . Subsequently, the related working hypothesis that the E1 Tor cell-bound haemagglutinin is playing an adhesive role is validated . In natural immunity, it could be envisaged that antiserum to this cell-bound haemagglutinin of V . cholerae E1 Tor would be highly protective due to the synergistic effects of the dual protective mechanisms in operation at the intestinal sites viz . a masking and agglutinating phenomenon. Infect Immun, 1981 Dec, 34(3), 895 - 9 Factors affecting phaeomelanin production by a melanin-producing (mel) mutant of Vibrio cholerae; Ivins BE et al.; In a previous study we isolated melanin-producing (mel) mutants of Vibrio cholerae and demonstrated that production of melanin during growth on solid media was stimulated by L-tyrosine and L-cysteine . In the studies reported here we analyzed factors that affected melanin production in liquid media and determined the abilities of radioactively labeled amino acids to serve as precursors for the formation of melanin by V . cholerae . Radioactivity from L-cysteine and from L-tyrosine was preferentially incorporated into partially purified melanin, providing further evidence for production of phaeomelanin by V . cholerae . Cuprous ions stimulated production of melanin by V . cholerae in defined liquid medium, but melanin formation was inhibited by high concentrations of L-cysteine or thiouracil . The inhibition of melanin formation by sulfhydryl compounds is most likely due to their ability to bind copper ions that are essential for tyrosinase activity. Infect Immun, 1981 Dec, 34(3), 702 - 7 Serum resistance and hemagglutination ability of marine vibrios pathogenic for fish; Trust TJ et al.; Representative strains of marine vibrios pathogenic for fish were shown to be resistant to the bactericidal activity of normal (nonimmmune) rainbow trout (Salmo gairdneri) serum, and loss of this resistance coincided with a marked reduction in virulence . Thermal lability and a requirement for Mg2+, but not for Ca2+, suggested that a mechanism for serum killing was the alternative complement pathway . In the case of Vibrio anguillarum, serum resistance was not coded for by the virulence plasmid pJM1 . Additional testing showed that these pathogenic vibrios were able to agglutinate a variety of eucaryotic cells and that selected strains agglutinated trout erythrocytes; however, a correlation between strain virulence and the ability to agglutinate fish erythrocytes was not apparent . Moreover, whereas mannose was found to inhibit the agglutinating activity of several strains, two of the high-virulence strains displayed a transient, mannose-resistant hemagglutinating activity . No relationship between the carriage of pJM1 by the V . anguillarum strains and hemagglutinating activity was demonstrable. Eur J Biochem, 1981 Dec, 120(3), 577 - 84 The function of the subunits of the fumarate reductase complex of Vibrio succinogenes; Unden G et al.; The membrane-bound fumarate reductase complex of Vibrio succinogenes catalyzes the reduction of fumarate by 2,3-dimethyl-1,4-naphthohydroquinone (dimethylnaphthohydroquinone) and consists of three different peptides (Mr 79,000, Mr 31,000 and Mr 25,000), the smallest of which is cytochrome b {Unden, G., Hackenberg, H . and Kroger A . (1980) Biochem . Biophys . Acta 591, 275-288} . The complex was cleaved with guanidinium chloride, the resulting subunits characterized and their functions within the complex investigated by reconstitutional experiments . 1 . The Mr-79,000 subunits catalyzed the reduction of fumarate by benzylviologen radicals as well as the oxidation of succinate by methylene blue, but not fumarate reduction by dimethylnaphthohydroquinone . 2 . The spectral and the redox properties of the isolated cytochrome b (Mr 25,000) were equivalent to those of the high-potential cytochrome b of the bacteria . The isolated cytochrome b had a midpoint potential of -15 mV and was reducible by dimethylnaphthohydroquinone in the absence of the other subunits . 3 . The Mr-31,000 subunit did not catalyze any of the reactions mentioned above . For the reduction of cytochrome b by succinate in the presence of the Mr-79,000 subunit, an amount of the Mr-31,000 subunit was required which was equimolar to cytochrome b . 4 . The activity of fumarate reduction by dimethylnaphthohydroquinone could be restored by coprecipitation of the three subunits . It is concluded that the fumarate reductase complex has two different sites, which are essential for its function in the phosphorylative electron transport of the bacterium . The site reacting with the substrates fumarate and succinate is situated on the Mr-79,000 subunit, and that reacting with dimethylnaphthohydroquinone is cytochrome b . The Mr-31,000 subunit mediates the electron transport between cytochrome b and the Mr-79,000 subunit. Can J Microbiol, 1981 Dec, 27(12), 1252 - 9 Adherence of Vibrio parahaemolyticus and its relation to pathogenicity; Iijima Y et al.; Cultured epithelial cells were used to investigate the adherence of Vibrio parahaemolyticus . Correlation between adherence in vitro and pathogenicity (colonization) were shown by experiments on the distribution of vibrios in the small intestine of guinea pigs and on the lethal activity of vibrios to mice . In vitro adherence of 32 strains of V . parahaemolyticus including Kanagawa phenomenon (KP) positive and KP-negative strains was studied . The effect of purified KP toxin and antiserum against KP toxin was also studied . Adherence was not related to KP . Adherence of V . parahaemolyticus was thought to depend on viability, since the decrease in the number of colony-forming units paralleled the decrease in the number of bacteria adhering to epithelial cells, and bacteria fixed with ethanol and formalin did not show adherence. Zh Mikrobiol Epidemiol Immunobiol, 1981 Dec, (12), 69 - 75 {Evaluation of the adhesion, colonization and enterotoxigenicity of Vibrio cholerae in the enteral infection of intact and passively immunized newborn animals}; Efremov VE et al.; V . eltor were introduced into 323 suckling rabbits through a gastro-duodenal tube and into 629 suckling mice either by the same method, or per annum by a new specially developed technique . Passive immunization was achieved in suckling mice by the preliminary introduction of antibacterial sera into the small intestine or by feeding them with the milk of female mice immunized at the end of pregnancy, and in suckling rabbits by feeding them with the milk of goats immunized by introducing vibrios through the tests of the udder . To evaluate the results, the accumulation of liquid in the intestine and the development of diarrhea were taken into account, quantitative inoculation of homogenized intestinal matter was made; vibrios on the epithelium of the villi in the small intestine were counted and the changes of enterocytes and the lamina propria, observed in light and electron microscopy, were considered . The results of the complex quantitative evaluation in intact suckling animals revealed that the enterotoxigenicity of vibrios was manifested after their adhesion to and later colonization on the intestinal epithelium . Passive antibacterial immunization suppressed their adhesion and colonization, which resulted in protection from the enterotoxigenic effect of vibrios and their elimination from the intestine. J Hyg (Lond), 1981 Dec, 87(3), 485 - 91 A study of the incidence of Vibrio parahaemolyticus in Malaysian shrimp undergoing processing for export; Cann DC et al.; The incidence of Vibrio parahaemolyticus in products of the Malaysian export shrimp processing industry was investigated through the stages from the catch to that of the cooked, peeled and frozen product . The organism was commonly found in freshly caught and landed shrimp, and could be detected by enrichment culture at all stages of processing . The number of V . parahaemolyticus in shrimp varied from nil to 4x10(4), and 19 of the 50 serotypes in the current antigenic scheme were found, O1-K38 and O1-K32 occurring most often . All the isolates were Kanagawa-negative; one strain was a sucrose-positive variant . The study indicated that specifications of 10(2) g-1 for V . parahaemolyticus in raw tropical shellfish are too stringent but that the Malaysian shrimp industry should be able to meet this requirement for cooked shrimp. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1981 Dec, 251(2), 213 - 22 A comprehensive study of environmental and human pathogenic Vibrio alginolyticus strains; Larsen JL et al.; An investigation comprising 49 environmental (VAE), 7 human isolates (VAH) and 3 type culture (VAT) strains of Vibrio alginolyticus showed a very high similarity among the strains . By primary isolation a swarming activity on blood agar and a yellow, hemispheric relatively large colony type on TCBS were very conspicuous . Besides the characteristics of Vibrionaceae the most typical and differential diagnostic important features were: sucrose fermentation, lysine decarboxylase activity, acetoine and 2,3 butanediol production, no growth on an electrolyte deficient medium (CLED), arginine and arabinose negative reaction . Swarming on blood agar and growth in 10% NaCl were also important although one of these criteria might be negative . An obvious difference was observed in ornithine where the percentage of positive VAE, VAH and VAT were 51, 100 and 0, respectively . In cellobiose all VAE and VAT but only two VAH were positive . MR positive strains were also found among all categories . The G + C mol% in the strains ranged from 45.9 - 46.8. J Clin Microbiol, 1981 Dec, 14(6), 631 - 9 Characterization of biochemically atypical Vibrio cholerae strains and designation of a new pathogenic species, Vibrio mimicus; Davis BR et al.; Biochemically atypical strains classified as Vibrio cholerae were characterized by biochemical reactions, serology, antibiotic susceptibility testing, and deoxyribonucleic acid relatedness . Strains with the following atypical reactions were shown to be V . cholerae: mannose negative, mannitol negative, lysine decarboxylase negative, no growth in the presence of 5% NaCl, salicin and cellobiose positive . Sucrose-negative strains were shown to constitute a new species, Vibrio mimicus, whose type strain is 1721-77 (ATCC 33653) . In addition to its negative sucrose reaction, V mimicus was differentiated from V . cholerae by its negative Voges-Proskauer, corn oil, and Jordan tartrate reactions and by its sensitivity to polymyxin . V . mimicus was isolated from shellfish and water, as well as from human diarrheal stools and ear infections . Most strains were typable with antisera against V . cholerae . Strains from three serogroups produced either a heat-labile or a heat-stable enterotoxin. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1981 Dec, 251(1), 70 - 8 Evaluation of a test-kit (Oxi/Ferm system) for identification of vibrios; Sanyal SC; A total of 258 strains belonging to different species of Vibrio isolated from various sources in different parts of the world were subjected to identification by Oxi/Ferm tube and conventional tests sometimes extending to 250 characters when the conventional tests failed to recognise them . Nearly 88 per cent of them could be identified accurately in this system at the species level . The strains designated as mannitol negative vibrios and Vibrio species were also identified properly as V . cholerae . However, the system recognised V . fluvialis (Group F vibrios) as Aeromonas hydrophila and some strains of L vibrios and V . parahaemolyticus as V . cholerae . Those strains that could be identified properly corresponded to one or more ID values of the kit . A few additional supplemental tests based on stable characters are recommended for inclusion to those ID values to identify all the vibrios accurately and in case of V . cholerae up to biotype level . Some of these tests are already included in the supplementary list of the system, the only change required is to put them against respective ID values . Reactions within the system were reproducible . This study establishes the utility of the convenient Oxxi/Ferm system for rapid and easy identification of the different members of the family Vibrionaceae including the newly recognised ones with few additional tests. Am J Clin Pathol, 1981 Dec, 76(6), 765 - 72 Histopathology of marine vibrio wound infections; Beckman EN et al.; Although marine vibrio wound infections and septicemia are being reported with increasing frequency, description of the histopathologic changes has been scanty . The histologic alterations in three patients with primary marine vibrio wound infections are presented . The lesions are characterized by intense acute cellulitis of the subcutis with much tissue destruction and extension into the adjacent dermis . The superficial dermis is devitalized and lacks an inflammatory cellular infiltrate . Subepidermal noninflammatory bullae are formed . Many organisms are seen both within the areas of intense acute inflammation and in devitalized areas . Organisms and inflammation are especially oriented around vessels, with associated acute vasculitis . It is concluded that the morphologic picture in marine vibrio wound infections is nonspecific yet characteristic. Arch Microbiol, 1981 Dec, 130(4), 276 - 80 Regulation of exoprotease production by temperature and oxygen in Vibrio alginolyticus; Hare P et al.; The production of an extracellular collagenase and alkaline protease by Vibrio alginolyticus during stationary phase was inhibited by a temperature shift from 30 to 37 degrees C and by a lack of oxygen . The stability of the exoproteases was unaffected by incubation at 37 degrees C and aeration . The optimum growth temperature for the V . alginolyticus strain was 33.5 degrees C and there was no difference in the growth rate at 30 and 37 degrees C . Aeration enhanced the rate of growth of exponential phase cells . Temperature and oxygen did not affect the growth of stationary phase cells when the exoproteases were being produced . Macromolecular synthesis in stationary phase cells was not affected by temperature . There was no rapid release of the exoproteases after temperature shift down and chloramphenicol inhibited the production of the enzymes when added at time of temperature shift down from 37 to 30 degrees C . The regulation of exoprotease production by temperature and oxygen was specific and has implications regarding the ecology of V . alginolyticus . Cerulenin, quinacrine and O-phenanthroline inhibited the production of the exoproteases. Am J Trop Med Hyg, 1981 Nov, 30(6), 1277 - 80 A Vibrio cholerae infection in a transient teamster; Holmes FF et al.; An illness indistinguishable clinically from classical cholera but caused by a non-cholera vibrio occurred in an over-the-road truck driver . The infecting organism was not finally identified as Vibrio cholera, Smith serotype 113 toxin positive, until 4 weeks after his hospital discharge . Hospital laboratories in most parts of the United States are unlikely to identify Vibrio cholerae in stool cultures unless specifically requested to do so . If one recognizes the chronic cholera carriers have been documented, that small epidemics do occur in unlikely places, and the lack of evidence that toxigenic V . cholerae 01 differs from toxigenic V . cholerae non-01 in these two respects, then the potential for epidemics in the United States is real. Zh Mikrobiol Epidemiol Immunobiol, 1981 Nov, (11), 41 - 4 {Taxonomic position of representatives of the species Vibrio cholerae}; Libinzon AE et al.; The data contained in the works of some foreign authors who have reconsidered the taxonomic position of V . metschnikovii are analyzed . In contrast to the views expressed by Gamaleya whi believed this microorganism to be the modified cholera vibrio, the foreign authors identify it with the "Kommabacillus der Cholera nostras" described by Finkler and Prior (1884) and classify it with saprophytes widely spread in the environment . These points of view were refuted by the data resulting from the study of the strains of V . metschnikovii and the Finkler - Prior vibrio from Soviet collections . The variant from the Odessa Institute of Epidemiology and Microbiology has been found to be most similar to the original characteristic given by Gamaleya and should be considered the lectotype of V . metschnikovii. Infect Immun, 1981 Nov, 34(2), 503 - 7 Role of iron in the pathogenesis of Vibrio vulnificus infections; Wright AC et al.; Infections with Vibrio vulnificus resulting in septicemia and high mortality have been correlated with pre-existing liver disease and hemochromatosis . As these conditions are associated with impaired iron metabolism and as iron availability in the host has been implicated in the pathogenicity of a number of bacterial infections, the role of iron as a possible factor in the pathogenesis of V . vulnificus was examined . Injection of mice with iron resulted in a lowering of the 50% lethal dose from 10(6) to 1.1 cells and in a reduction in the time of death postinfection . Elevated serum iron levels were also produced by damaging livers with injections of CCl4 . The inoculum size required to kill these mice was directly correlated with serum iron levels . Since the portal of infection of this organism may be ingestion of contaminated seafood, the effects of iron upon orally induced infection were also studied . The effects of adding iron, transferrin, or Desferal (an iron chelate) upon the growth of V . vulnificus in human and rabbit sera were also examined . Iron appeared to be the limiting factor in the ability of this organism to survive or grow in mammalian sera . These results, both in vitro and in vivo, provided strong evidence that iron may play a major role in the pathogenesis of V . vulnificus. Hoppe Seylers Z Physiol Chem, 1981 Nov, 362(11), 1507 - 14 Phagocytosis of sialidase-treated rat erythrocytes: evidence for a two-step mechanism; Kuster JM et al.; Binding and phagocytosis of rat erythrocytes by liver and peritoneal macrophages were studied with a radioactive in vitro assay which yields quantitative data . Partial removal of sialic acids from the erythrocytes by Vibrio cholerae sialidase resulted in a marked increase of binding of the red cells by both liver and unstimulated peritoneal macrophages . Peritoneal macrophages stimulated by thioglycolate or starch, however, did not differentiate between control and desialylated erythrocytes . By inhibition experiments it was confirmed that rat peritoneal macrophages bind homologous sialidase-treated erythrocytes via a beta-D-galactose-specific lectin on the macrophage surface . While this attachment already occurs in buffer, serum was required for the subsequent phagocytosis . The possible involvement of factors of the complement system in the phagocytosis step was evidenced by a marked decrease of phagocytosis after heat inactivation of the serum . Based on these experiments, we propose a model of a two-step mechanism for the uptake of sialidase-treated erythrocytes by macrophages, comprising both the lectin and a receptor for serum components. Chem Biol Interact, 1981 Nov, 37(3), 321 - 35 Radiomimetic property of furazolidone and the caffeine enhancement of its lethal action on the vibrios; Banerjee SK et al.; Sensitivities of the strains belonging to four vibrio biotypes to the action of furazolidone were investigated . Vibrio cholerae (classical) was most and Vibrio parahaemolyticus least sensitive to this drug . Statistical analyses revealed significant differences between any two of the four types of vibrio in respect of their sensitivity to furazolidone . The drug was radiomimetic in action, the doses of UV light (DUV) and furazolidone (Df) required for 10% survival of the vibrios being correlated by the equation, Df = 0.28 exp . (0.008 DUV) . Caffeine exhibited lethal synergism with furazolidone and the synergistic effect depended on the mode of caffeine treatment, the effect being maximum when caffeine was present along with and also after furazolidone treatment . UV spectrophotometric study revealed that caffeine did not bind with native DNA but did so with denatured DNA resulting in a bathochromic shift and a quenching of the caffeine absorption maximum at 209.4 nm . The binding isotherm (Scatchard plot) indicated the presence of a heterogeneity in the binding sites and that the parameters for the strongest mode of bonding were n = 0.254 and k = 7.5 X 10(5) M-1. Zh Mikrobiol Epidemiol Immunobiol, 1981 Nov, (11), 31 - 6 {Isolation and characteristics of E . coli plasmid-determined K88 antigen}; Vodop'ianov SO et al.; The advantage of ultracentrifugation over isoelectric focusing in the isolation of antigen K88 from E . coli is shown . The production of antigen K88 is recA-independent . The antigen has been found to be nontoxic for mice, to have the isoionic point at pH 4.1 and the sedimentation coefficient 16.6S, to completely consist of protein subunits of 25,000 daltons . The expression of antigen K88 enhances the adhesive properties of the host cell . No relationship between antigen K88 and the supposed adhesins of both Vibrio cholerae biotypes has been established. J Gen Microbiol, 1981 Nov, 127, pt 1, 193 - 9 Regulation of extracellular alkaline protease activity by histidine in a collagenolytic Vibrio alginolyticus strain; Long S et al.; Vibrio alginolyticus synthesized an inducible extracellular collagenase in a peptone medium during the stationary growth phase . These cultures also possessed extracellular alkaline serine protease activity . The alkaline protease activity did not require a specific inducer and it was produced in tryptone or minimal media . The collagenase was not produced in either the tryptone or minimal media . The alkaline protease activity was sensitive to catabolite repression by a number of carbon sources, including glucose, and by amino acids and ammonium ions . Cyclic AMP, dibutyryl cyclic AMP and cyclic GMP did not relieve catabolite repression . Histidine and urocanic acid stimulated the production of alkaline protease activity in tryptone and minimal media . Other compounds associated with the histidine utilization (hut) pathway did not increase alkaline protease activity . Histidine reversed the repression of alkaline protease activity by glucose of (NH4)2SO4 in minimal medium . Histidine and the compounds associated with the hut pathway inhibited collagenase production. Proc Natl Acad Sci U S A, 1981 Nov, 78(11), 6958 - 62 Induction by cholera toxin of synchronous divisions in vivo in the epidermis resulting in hyperplasia; Kuroki T; Intracutaneous injection of cholera toxin, exotoxin of Vibrio cholerae, into the dorsal skin of mice, rats, and hamsters at doses of greater than 0.1 ng evoked an acute reaction at the site of injection, which was characterized histologically by an edematous reaction in the dermis and mitotic stimulation in the epidermis . Mitotic and labeling induces of basal cells of the mouse epidermis showed two peaks at 24 and 48 hr after injection, thereby producing epidermal hyperplasia . The thickness of the intrafollicular epidermis increased progressively from 32 hr after toxin injection, being greatest on day 4 and decreasing to normal on day 7 . The epidermis on day 4 after injection of 1.0 ng of toxin was about 4- to 6-fold thicker than normal or phosphate buffer-treated control skin . This sequence of events indicated that cholera toxin induced two successive synchronous divisions of the epidermal cells and produced temporary hyperplasia without interfering with epidermal differentiation . The complete structure and function of the cholera toxin are required for induction of epidermal hyperplasia: no mitotic stimulation was induced by injection of the A and B units of the cholera toxin molecule or by preincubation of the toxin with anti-cholera toxin antibody and with the membrane receptor, GM1 ganglioside . Five other agents known to increase the level of intracellular cyclic AMP by different means (dibutyryl cyclic AMP, 3-isobutyl-1-methylxanthine, theophylline, isoproterenol, and prostaglandin E1) did not produce a skin reaction. Infect Immun, 1981 Nov, 34(2), 333 - 6 Isolation of special antibodies which react only with homologous enterotoxins from Vibrio cholerae and Enterotoxigenic Escherichia coli; Honda T et al.; Cholera enterotoxin (CT) and heat-labile enterotoxin (LT) produced by enterotoxigenic Escherichia coli share a common antigenic determinant . In addition, CT and LT each have a unique antigenic determinant . Antisera were prepared by immunoaffinity chromatography against these unique antigenic determinants, that is, antiserum that reacted with CT but not with LT and antiserum that reacted with LT but not with CT . Antiserum against the common antigenic determinant to CT and LT was also prepared by immunoaffinity chromatography . The specificities of these antisera were demonstrated by Ouchterlony double-gel diffusion tests and by neutralization of the activities of CT and LT to cause morphological changes in Chinese hamster ovary cells. Biochim Biophys Acta, 1981 Oct 27, 655(3), 413 - 20 Repair of ultraviolet-light-induced DNA damage in vibrio cholerae; Das G et al.; Repair of ultraviolet-light-induced DNA damage in a highly pathogenic Gram-negative bacterium, Vibrio cholerae, has been examined . All three strains of V . cholerae belonging to two serotypes, Inaba and Ogawa, are very sensitive to ultraviolet irradiation, having inactivation cross-sections ranging from 0.18 to 0.24 m2/J . Although these cells are proficient in repairing the DNA damage by a photoreactivation mechanism, they do not possess efficient dark repair systems . The mild toxinogenic strain 154 of classical Vibrios presumably lacks any excision repair mechanism and studies of irradiated cell DNA indicate that the ultraviolet-induced pyrimidine dimers may not be excised . Ultraviolet-irradiated cells after saturation of dark repair can be further photoreactivated. Biochim Biophys Acta, 1981 Oct 13, 661(2), 295 - 302 Iron-sulphur clusters in fumarate reductase from Vibrio succinogenes; Albracht SP et al.; (1) The fumarate reductase complex from Vibrio succinogenes contains one FAD molecule, one {4Fe-4S}3+(3+,2+) and one {2Fe-2S}2+(2+,1+) cluster per enzyme molecule . Both clusters can be partly reduced by succinate . In the presence of excess Na2S2O4 and fumarate, the {2Fe-2S} cluster is completely oxidized, whereas the other cluster is largely reduced . (2) The {2Fe-2S} cluster is localized in the Mr, 31,000 subunit . The EPR spectrum of the reduced cluster in the isolated subunit differs slightly in line width, but not in g-value, from the spectrum of reduced, intact enzyme complex . The demonstrates that the immediate environment of th cluster is little perturbed by dissociating this subunit from the FAD-containing Mr 79,000 subunit . The temperature dependence of the power-saturation behaviour has, however, greatly decreased in the isolated subunit, the saturation at 11 K of the paramagnetic cluster being much less than in the enzyme complex . Moreover, the temperature dependence of th power-saturation behaviour of this cluster in the enzyme is greater with succinate as reducing agent, than with dithionite . (3) The {4Fe-4S} cluster is located on the Mr 79,000 subunit . This cluster is unstable in air when the subunit has been dissociated from the enzyme complex. J Wildl Dis, 1981 Oct, 17(4), 521 - 7 Mortality in captive Atlantic cod, Gadus morhua, associated with fin rot disease; Khan RA et al.; A total of 320 of 621 (52%) captive Atlantic cod, Gadus morhua, died from fin rot over an 8-year period; mortality reached high proportions in 1979 when 110 of 168 (66%) succumbed . Lesions were confined to the fins, skin and dermal musculature and were observed 3-10 days after the fish were placed in laboratory aquaria . Erosion of the fins and caudal peduncle, accompanied by petechiae and ulceration also were apparent in the trunk region . Most deaths occurred within 2 months after capture . Infection was associated with depressed hematocrit, hemoglobin and total plasma protein and an increase in circulating immature erythrocytes and neutrophils . Mortality was probably due to physiological stress resulting from excessive blood loss . Three genera of bacteria, mostly Pseudomonas, but also Aeromonas and Vibrio, were isolated from fin rot tissues and possibly are the causative agents of the disease. Can J Microbiol, 1981 Oct, 27(10), 1048 - 52 Polyacrylamide gel electrophoresis and infrared spectroscopy of vibrio biotypes; Maiti M et al.; Polyacrylamide gel electrophoresis and infrared spectroscopy were used to study the relationship between Vibrio cholerae (classical), Vibrio cholerae (El Tor), and nonagglutinable (NAG) vibrios . Acid extracts of the different strains produced type-specific electrophoretic patterns, and the infrared spectra revealed broad absorption maxima which largely correspond to those found in other organisms . With the exception of the NAG vibrios, the infrared spectra of cholera El Tor vibrios were identical . Strain-specific differences were found in the exoprotein spectra of these organisms by the sodium dodecyl sulphate - polyacrylamide gel electrophoretic technique. Infect Immun, 1981 Oct, 34(1), 90 - 7 Production of cholera-like enterotoxin by a Vibrio cholerae non-O1 strain isolated from the environment; Craig JP et al.; Vibrio cholerae non-O1 strain E8498, isolated in 1978 from fresh water in Louisiana, produced a vascular permeability factor when cultured in shallow resting cultures of Casamino Acids-yeast extract-glucose medium for 24 h at 30 degrees C . Undiluted resting culture filtrates contained heat-labile permeability factor activity which was only partially neutralized by cholera antitoxin and GM1 ganglioside . Supernatants concentrated with PM-10 membranes caused hemorrhage and necrosis in rabbits within 1 h after intracutaneous injection, whereas appropriate dilutions of both filtrates and concentrates demonstrated delayed permeability factor activity, without hemorrhage or necrosis, which was indistinguishable in appearance from that caused by purified cholera enterotoxin produced by V . cholerae O1 Inaba strain 569B . Crude E8498 filtrates contained the biological equivalent of about 5 ng/ml of purified enterotoxin . Permeability factor activity in the fraction obtained by 20 to 50% saturation of filtrate concentrate with ammonium sulfate could be completely neutralized by reference standard cholera antitoxin prepared against purified 569 B enterotoxin . Hemorrhagic activity was unaffected by cholera antitoxin . A 5,000-fold concentrate of the culture supernatant yielded a line of identity with purified cholera enterotoxin in an agar gel double-diffusion test against cholera antitoxin purified by affinity column chromatography with BrCN-activated Sepharose 4B-linked purified cholera enterotoxin as the adsorbent . These findings indicate that V . cholerae non-O1 E8498 produces a permeability factor which is immunologically and biologically indistinguishable from that produced by a strain of V . cholerae O1 classical biotype. Infect Immun, 1981 Oct, 34(1), 241 - 9 Role of chemotaxis in the association of motile bacteria with intestinal mucosa: in vitro studies; Freter R et al.; Various Sephadex G-15 fractions of pepsin-digested mucosal extract inhibited the in vitro association of cholera vibrios with mucosal slices . Inhibitory activity paralleled the taxin activity of the fractions for these bacteria . This supports the theory that inhibition of mucosal association by pepsin-digested mucosal scrapings was due to the blocking of taxin receptors on the bacterial surface . Nonchemotactic mutants were significantly less efficient than the chemotactic parent or revertant strains in associating with mucosal slices in vitro . Control experiments in which filter paper disks replaced the mucosal slices showed a comparable extent of association of chemotactic and nonchemotactic vibrios with this material . Histological studies indicated that vibrios associated predominantly with the mucus gel of the intestinal slices rather than with the mucosal epithelium or the serosal surface . Intestinal slices attracted chemotactic vibrios even after prolonged washing, suggesting continuous production of the taxin by the tissue . Inert polystyrene particles 1.1 micrometers in diameter penetrated the mucus gel of intestinal slices very poorly, but nevertheless could be detected in low numbers in the deep intervillous spaces within 15 min . In contrast, chemotactic vibrios reached the deep intervillous spaces in significantly higher numbers, whereas motile, non-chemotactic vibrios reacted like the inert particles . It is concluded that mucus gel represents a partial barrier to the penetration of particles of bacterial size and that this barrier can be invaded efficiently by motile bacteria, but only when their locomotion is guided by chemotactic stimuli. Infect Immun, 1981 Oct, 34(1), 234 - 40 Role of chemotaxis in the association of motile bacteria with intestinal mucosa: in vivo studies; Freter R et al.; In vivo loops were prepared in the small intestine of rabbits and injected with mixtures of Vibrio cholerae and polystyrene spheres (1.1-micrometers diameter) . The loops were removed and frozen after 15 min and then sectioned in a cryostat . The locations of particles and vibrios were determined microscopically . The vibrio/particle ratio was unity in the lumen of the loops, but increased 10-fold in the deep intervillous spaces, indicating active invasion of the mucus gel by the chemotactic parent strain . Motile nonchemotactic mutants and nonmotile mutants of this strain invaded the mucus at the same rate as inert particles . Similar results were obtained with intestinal loops prepared in germfree mice . When germfree mice were disassociated with mixtures of chemotactic (parent or revertant) and nonchemotactic mutant vibrios in equal proportions, the chemotactic strain rapidly outgrew its nonchemotactic counterpart in the intestine . Nonchemotactic mutants introduced as monoassociates into germfree mice were rapidly overgrown by nonmotile mutants which apparently arose spontaneously in the gut . Motility was therefore beneficial to survival only when it was directed by chemotactic stimuli, whereas it was a liability in the absence of such stimuli . Growth of chemotactic vibrios in small intestinal loops of rabbits paralleled that of nonchemotactic mutants for the first 4 to 6 h . Thereafter, the growth rate of the chemotactic vibrios was significantly faster . This was correlated with a significantly higher degree of association with the mucosa on the part of the chemotactic vibrios. Infect Immun, 1981 Oct, 34(1), 222 - 33 Role of chemotaxis in the association of motile bacteria with intestinal mucosa: fitness and virulence of nonchemotactic Vibrio cholerae mutants in infant mice; Freter R et al.; Contrary to earlier findings with all other in vivo and in vitro models of cholera studied, nonchemotactic vibrio mutants showed a relatively greater fitness in 5-day-old infant mice as compared with chemotactic parent or chemotactic revertant strains . This trend was manifest in the relatively greater number of nonchemotactic mutants recovered from the upper small intestine at 4 and 18 h after intragastric infection . The same trend was also revealed in the significantly greater virulence (in terms of time to death) of nonchemotactic mutants as compared with the chemotactic parent or revertant strains . Histological studies in infant mice of the penetration of chemotactic and nonchemotactic vibrios into the mucus gel of the small intestine yielded the same findings as in all other models studied, i.e., significantly greater penetration by chemotactic vibrios . There was no correlation between the relative fitness of nonchemotactic vibrios in the small intestine of infant mice and the rate of recovery of viable nonchemotactic vibrios from that site . In contrast, excellent correlation was found between the relative fitness of nonchemotactic vibrios and a decrease in the recovery of viable cells of the chemotactic strain from the small intestine . This indicates that the relatively greater fitness of the nonchemotactic vibrios in infant mice was only apparent and that the observed phenomenon was actually due to an antibacterial mechanism which prevented the accumulation of the chemotactic strains in the small intestine rather than to any stimulating effect on the nonchemotactic mutant itself . To study the in vivo fate of the inoculum in infant mice, vibrios were labeled with either 32P, 35S, or {3H}thymidine . Specific activity determinations of the 32P label were compatible with the assumption of an accelerated rate of death of the chemotactic parent strain in the small intestine . Results with the other isotopes, however, were significantly different . Indeed, the amount of radioactivity retained in the small intestine after feeding labeled bacteria correlated more closely with the isotope used than with the strain of vibrio under study . Consequently, considerable doubt must be cast on the general validity of this not uncommon technique for determining the in vivo location and the death or survival of radioactively labeled bacteria. Infect Immun, 1981 Oct, 34(1), 215 - 21 Role of chemotaxis in the association of motile bacteria with intestinal mucosa: chemotactic responses of Vibrio cholerae and description of motile nonchemotactic mutants; Freter R et al.; A motile, chemotactic, Ogawa strain of Vibrio cholerae was attracted by all 20 L-amino acids tested, in contrast to Escherichia coli AW 405, which did not react to several of these . The maximum number of vibrios entering a capillary was much lower when the capillary contained carbohydrates rather than amino acids, but the minimum effective concentrations of the carbohydrates and amino acids tested were of the same order of magnitude . L-Fucose, a sugar known to inhibit the adhesion of this vibrio strain to brush border membranes, had no attraction (taxin activity) for it . A pepsin digest of rabbit mucosal scrapings or tryptone attracted vibrios as strongly as the most active amino acids . Several nonchemotactic and one nonmotile mutant were selected from the parent vibrio . The nonchemotactic mutants were indistinguishable from the parent in their ability to attach in vitro to isolated intestinal brush border membranes, whereas the nonmotile mutant had lost this ability . Parent and nonchemotactic mutants had equal growth rates in stirred and still continuous flow cultures that were maintained in an anaerobic environment. J Clin Microbiol, 1981 Oct, 14(4), 457 - 9 Hemolytic reaction of clinical and environmental strains of Vibrio vulnificus; Johnson DE et al.; All Vibrio vulnificus strains tested (four clinical isolates and eight environmental isolates) hemolyzed human erythrocytes . In contrast to findings with Vibrio parahaemolyticus, in which hemolytic activity correlates with isolation from clinical specimens, results from the present study suggest that hemolysis is not usefull in differentiating V . vulnificus strains. Infect Immun, 1981 Oct, 34(1), 115 - 9 Effect of pH on the production of the Kanagawa hemolysin by Vibrio parahaemolyticus; Cherwonogrodzky JW et al.; Production of the Kanagawa hemolysin by patient strains of Vibrio parahaemolyticus was found to respond to the pH rather than to the type of carbohydrate present in the growth medium . Regardless of the carbohydrate present, hemolysin production in peptone broth cultures occurred only when the pH was between 6.5 and 5.5 . Mannitol, the sugar used in the Wagatsuma agar, lowered the pH to within this range, thus providing optimal conditions for hemolysin production . Glucose and mannose, although readily metabolized, lowered the pH below this range, inhibiting growth and hemolysin production . Alkaline cultures either without carbohydrates or containing non-metabolizable sugars showed little hemolytic activity because the pH always remained alkaline . In pH-stat cultures maintained at pH 6.2, higher hemolysin yields were produced irrespective of the presence or absence of mannitol . We conclude that the production of the Kanagawa hemolysin is under pH control . Marine strains of V . parahaemolyticus, which are Kanagawa negative, did not express detectable amounts of hemolysin under those conditions shown to stimulate hemolysin production by Kanagawa-positive strains. J Bacteriol, 1981 Oct, 148(1), 374 - 8 Mutagenesis by insertion of drug resistance transposon Tn7 into a vibrio species; Thomson JA et al.; A halotolerant, collagenolytic strain of Vibrio sp . was conjugated with an Escherichia coli strain carrying plasmid RP4 . The plasmid was transferred to and maintained in the Vibrio and could be subsequently transferred in matings to suitably marked stains of the same species . After conjugation with an E . coli carrying the cointegrate plasmid RP4::Mu cts61::Tn7, Vibrio transconjugants were selected that carried Tn7 inserted into the bacterial chromosome . A large proportion of these transconjugants were auxotrophic, showing that plasmid suicide by Mu can be used to isolate Tn7-derived mutants in Vibrio . Approximately half of the auxotrophs isolate Tn7-derived mutants in Vibrio . Approximately half of the auxotrophs isolated were ilv mutants, all of which exhibited the same phenotype . Thus, although Tn7 insertion can induce auxotrophy, including trp, thy, his and ura, in Vibrio, there does appear to be a hot spot for integration in the ilv operon. J Infect Dis, 1981 Sep, 144(3), 244 - 8 Interaction of Vibrio vulnificus with human polymorphonuclear leukocytes: association of virulence with resistance to phagocytosis; Kreger A et al.; The results of studies described in this report support the ideas that virulence of Vibrio vulnificus is associated, at least in part, with resistance to phagocytosis and that the ability of the bacterium to resist phagocytosis results from its possession of an antiphagocytic surface antigen . These conclusions are based on the observations that (1) human polymorphonuclear leukocytes (PMNLs) exhibited a significantly weaker chemiluminescent response and phagocytic response when interacting with a virulent strain of the bacterium than when challenged with a weakly virulent strain of the bacterium; (2) rabbit antiserum to the virulent bacterium, but not normal rabbit serum, significantly enhanced the chemiluminescent response of PMNLs challenged with the virulent bacterium and significantly enhanced ingestion of the bacterium by the PMNLs; and (3) the opsonic activity of the antiserum was removed by adsorption with a formalin-killed, whole cell preparation of the virulent bacterium. J Biochem (Tokyo), 1981 Sep, 90(3), 619 - 28 Partial purification and properties of respiratory chain-linked l-glycerol 3-phosphate dehydrogenase from a marine bacterium, Vibrio alginolyticus; Unemoto T et al.; Respiratory chain-linked L-glycerol 3-phosphate (G3P) dehydrogenase {EC 1.1.99.5} of marine bacterium, Vibrio alginolyticus, was extracted from the membrane fraction by treatment with Tween 20, and fractionation on DEAE-Sephacel and QAE-Sephadex in the presence of 0.05% Liponox DCH(alkyl polyoxyethylene ether) yielded a preparation having a specific activity of 22.1 units/mg protein when assayed by phenazine methosulfate (PMS)-coupled reduction of thiazolyl blue tetrazolium (MTT) . The purified enzyme had an apparent molecular weight of 300,000 as determined by chromatography on Sepharcyl S-300 in 0.05% Liponox DCH, and had noncovalently bound FAD as its coenzyme . The enzyme had a pH optimum of 8.5-9.0, and required 200 mM NaCl or KCl and an appropriate detergent (such as Tween, Brij or Liponox DCH) for maximum activation . The activating effect of NaCl was due to a decrease in Km for G3P and that of Tween 20 was due to both a decrease in Km and an increase in Vm . Triton X-100 could not activate the enzyme and was inhibitory in the presence of phospholipids . The reaction followed a ping-pong mechanism . IN addition to PMS, 2,6-dichlorophenol indophenol and duroquinone, the enzyme could reduce ubiquinone-5 (Q-5) in the presence of Liponox DCH at a rate of 46% of the PMS reductase activity . The enzyme was strongly inhibited by heavy metal ions and by p-chloromercuribenzoate . The activity for Q-5, but not for PMS, was inhibited by o-phenanthroline and bathophenanthroline, suggesting the participation of nonheme iron protein in the Q-5 reduction. Proc Natl Acad Sci U S A, 1981 Sep, 78(9), 5406 - 10 Specific gangliosides function as host cell receptors for Sendai virus; Markwell MA et al.; The ability of specific gangliosides to function as host cell receptors for Sendai virus was investigated by using Madin-Darby bovine kidney cells which become resistant to infection upon treatment with Vibrio cholerae sialidase . Sialidase-treated cells were incubated for 20 min at 37 degrees C with individual, highly purified gangliosides containing homogeneous carbohydrate moieties and then inoculated with virus for 10 min . Susceptibility of the cells to infection was monitored by hemagglutination titer of the virus produced 48 hr after inoculation . Incubation of the cells with gangliosides containing the sequence NeuAc alpha 2,3Gal beta 1,3GalNAc (i.e., GD1a, GT1b, and GQ1b) fully restored susceptibility to infection to the cells . However, the ganglioside GQ1b in which the sequence ends with two sialic acids in a NeuAc alpha 2,8NeuAc linkage instead of a single sialic acid as in GD1a and GT1b, was effective as a receptor at a concentration 1/100th that of any of the other gangliosides tested . Incubation with gangliosides similar in structure to GD1a, GT1b, and GQ1b but lacking the sialic acid attached to the terminal galactose (i.e., GM1 and GD1b) had no effect . The results from control experiments in which gangliosides were incubated at 0 degrees C with cells or in which trypsin was used to remove gangliosides adsorbed to cells were consistent with the premise that the gangliosides must actually insert into the cellular membrane to function as Sendai virus receptors . Addition of 4 X 10(6) molecules of 14C-labeled GD1a per cell made the cells fully susceptible to infection . Analysis of the ganglioside content of cell membranes showed that gangliosides GD1a, GT1b, and GQ1b are natural components of these cells and are present in quantities sufficient to act as receptors . These results demonstrate that gangliosides with the proper carbohydrate sequence, such as GD1a, GT1b, and GQ1b, function as natural receptors for Sendai virus in host cells. J Biol Chem, 1981 Aug 25, 256(16), 8357 - 63 Different cell-surface receptor determinants of antigenically similar influenza virus hemagglutinins; Carroll SM et al.; Two influenza virus substrains, A/RI/5-/57 and A/RI/5+/57, with antigenically similar hemagglutinins and neuraminidases but with different properties of elution from erythrocytes, have been examined for the specificity of their interaction with cell surface sialyloligosaccharides . This was accomplished by using erythrocytes treated with Vibrio cholerae neuraminidase to remove sialic acids, and then modified with CMP-NeuAc and three purified sialyltransferases to contain either the NeuAc alpha 2,3Gal, NeuAc alpha 2,6Gal, or NeuAc alpha 2,6GalNAc linkages on cell-surface glycoproteins . Each virus was tested for its ability to adsorb to and hydrolyze sialic acid from the derivatized cells . The hemagglutinins of the A/RI/5-/57 and A/RI/5+/57 viruses were found to have totally different specificities, binding respectively to the NeuAc alpha 2,3Gal and NeuAc alpha 2,6Gal linkages as preferred receptor determinants . In contrast, the neuraminidases of the two viruses exhibited similar specificities, efficiently hydrolyzing only the NeuAc alpha 2,3Gal linkage . The results suggest that the difference in the ability of the A/RI/5-/57 and A/RI/5+/57 viruses to elute from erythrocytes resides in the different receptor specificities of their hemagglutinins . Indeed, it appears that A/RI/5-/57 virus elutes because its receptor determinant, NeuAc alpha 2,3Gal, is rapidly hydrolyzed by the viral neuraminidase, while the A/RI/5+/57 virus fails to elute because its preferred receptor determinant NeuAc alpha 2,6Gal is largely resistant to hydrolysis. Lancet . 1981 Aug 22;2(8243):419. Breast milk and infection; Jelliffe DB et al.; PIP: There were several problems with the editorial on breastmilk and infection appearing in the May 30 Lancet . Of the antiinfective substances in human milk, lysozyme was omitted, as were the bifidus factor, the antistaphylococcus factor, antitoxins for neutralizing Vibrio cholerae and Escherichia coli, lactoperoxidase, and volatile fatty acids . Low pH and the exclusive specific bifidus bowel flora are all components of the same interacting antiinfective screen, as are macrophages and lymphocytes . Secretory IgA in breastmilk as a developmental bridging mechanism until the infant's intestine can secrete IgA itself at 3 months, is not recognized . The protective effect of human milk against death from diarrheal disease is well-known both in Europe and North America and in such disadvantaged communities as exist in the 3rd world . Clearly, this is related in part to a lack of contamination of human milk; the process is both passive and active . Recent studies show protective effects against infectious episodes in babies in well-to-do communities . The role of colostrum as an immunological bolus for the newborn is barely mentioned despite long known high levels of antibodies and the probability that neonatal diarrhea is often a colostrum deficiency syndrome . Doses of colostrum (5 mg/kg daily) reduce the incidence of endemic nursery neonatal E . coli diarrhea . The protective effects extend to other extraintestinal infections and to certain viral infections . There is no mention of the dyadic immunological interaction between mother and young baby, particularly through the proven "gut-mammary axis" . The editorial concludes by damning with faint praise . For example, it is outdated, negative editorial revealing scant knowledge of the considerable published literature which documents the protective effect of breastfeeding against many alimentary and extraintestinal infections . author's modified Am J Epidemiol, 1981 Aug, 114(2), 293 - 8 Non-O group 1 Vibrio cholerae gastroenteritis associated with eating raw oysters; Wilson R et al.; A cluster of five cases of non-O group 1 (non-O1) V . cholerae gastroenteritis occurred in one Florida locality during November 1979 . Clinical findings included nausea, vomiting, and abdominal cramping in all affected persons; two had bloody diarrhea . All five persons gave a history of eating raw oysters within four days of onset of illness . A case-control study statistically associated the eating of raw oysters with development of illness (p = 0.0008); this finding was confirmed by a retrospective cohort study of patients hospitalized for diarrhea (p = 0.0001) . Non-O1 V . cholerae organisms were isolated from oysters and water samples taken from areas where ill persons had obtained their oysters . In at least one instance the same serotype was isolated from a patient's stool specimen and from the water where the patient had obtained oysters . Non-O1 V . cholerae infection must be considered in the differential diagnosis of shellfish-associated gastroenteritis. J Clin Microbiol, 1981 Aug, 14(2), 222 - 4 FK phage for differentiating the classical and El T or groups of Vibrio cholerae; Takeya K et al.; A new vibrio-infecting phage (FK phage) isolated from sewage lysed all strains of Vibrio cholerae biovar cholerae, whereas all strains of V . cholerae biovar El Tor were resistant to it . FK phage was entirely different from Mukerjee group IV phage in morphology and antigenicity . In addition to group IV phage, the use of FK phage will be useful in the examination and typing of V . cholerae. Zentralbl Bakteriol A, 1981 Aug, 249(3), 392 - 9 Experimental studies on enteropathogenicity and pathogenesis of group 'F' vibrio infections; Agarwal RK et al.; Group 'F' vibrios are a recently recognised group of bacteria that have been isolated from cases of diarrhoea and a variety of sources in the environment . They have been shown to cause accumulation of fluid in rabbit gut loops . The enterotoxic factor was found to be heat-labile as evidenced by heat treatment, time course of fluid accumulation and negative suckling mouse assay . It was associated with cytotoxic factor(s) . These organisms were seen to be lacking in invasive capacity . Prostaglandins and 5-hydroxytryptamine were found to play no role in causation of fluid outpouring into the loops but chlorpromazine was found to inhibit the fluid outpouring completely thereby indicating that the enterotoxin may act through mediation of cyclic AMP. J Med Microbiol, 1981 Aug, 14(3), 243 - 50 The role of trace elements and phosphates in the synthesis of vascular-permeability factor by Vibrio cholerae; Sagar IK et al.; Trace element and phosphate requirements for the synthesis of vascular-permeability factor (PF) by Vibrio cholerae strains B1307 and VC12 were investigated . While magnesium appeared to be indispensable for strain VC12, small amounts of PF were synthesised by strain B1307 in the presence of iron, zinc and manganese . However, even in the latter strain, maximum synthesis was recorded only in magnesium-containing media . Phosphates in the range 0.75-6.00mM controlled the synthesis of PF by both strains. Cancer Res, 1981 Aug, 41(8), 3082 - 6 Immunotherapy of L1210 leukemia using neuraminidase-modified plasma membranes combined with chemotherapy; Pincus JH et al.; Purified L1210 plasma membranes treated with Vibrio cholerae neuraminidase (VCN) were used for active immunotherapy of L1210 tumors in DBA/2J mice . Immunotherapy with VCN-treated membranes was effective only when combined with 1-(2-chloroethyl)-3-(4-methylcyclohexyl)-1-nitrosourea (MeCCNU) . Successful therapy was a function of the dose of MeCCNU, the dose of VCN-treated membranes, and the time after MeCCNU treatment when VCN-treated membranes were administered . Optimum conditions for treating animals with tumors initiated with 10(4) cells were MeCCNU (20/kg) given 3 days after tumor inoculation and 0.25 mg VCN-treated membranes given 1 day after chemotherapy . Control membranes, not treated with VCN, that were administered 1 day after MeCCNU were ineffective; when given 4 days after chemotherapy, the caused accelerated mortality, suggesting immunological enhancement of tumor growth . Our results indicate that VCN-treated plasma membranes can be used for active immunotherapy of established tumors and underscore the importance of carefully designing immunotherapy protocols to achieve optimum desirable effects. Cancer Res, 1981 Aug, 41(8), 3077 - 81 Characterization and use of neuraminidase-modified L1210 plasma membranes for protection against tumor growth; Brandt AE et al.; Purified plasma membranes were prepared from L1210 ascites tumor cells and analyzed for their protein and carbohydrate composition . Conditions were developed for treating the isolated plasma membranes with Vibrio cholerae neuraminidase (VCN) so that 88% of the N-acetylneuraminic acid was removed without changing membrane proteins or other membrane carbohydrate constituents . The VCN-induced modifications were characterized by labeling VCN-treated and untreated L1210 cells by the galactose oxidase:sodium {3H}borohydride procedure . This showed that N-acetylneuraminic acid is the predominant saccharide at the nonreducing terminus of plasma membrane glycoproteins and that galactose and/or N-acetylgalactosamine residues are penultimate to these . VCN modification exposed the penultimate residues and was not limited to any single plasma membrane glycoprotein . DBA/2J mice were given i.p . injections of VCN-treated or untreated membranes and were challenged 3 weeks later with 10(4) viable L1210 cells . Mice pretreated with VCN-treated membranes resisted the tumor challenge; those receiving untreated membranes or no treatment succumbed to the tumor . Our results demonstrate that appropriately modified plasma membranes can be used to induce resistance to tumor growth . They also suggest that tumor cell membrane carbohydrate structures have an important role in this phenomenon. Infect Immun, 1981 Aug, 33(2), 583 - 90 Detection of extracellular toxin(s) produced by Vibrio vulnificus; Kreger A et al.; Conditions are described for the production, in high titers, a heat-labile, antigenic, extracellular toxin(s) by Vibrio vulnificus, a recently recognized human pathogen . Bacteriologically sterile culture filtrate preparations obtained from mid-logarithmic-phase cultures of the bacterium possessed cytolytic activity against mammalian erythrocytes, cytotoxic activity for Chinese hamster ovary cells, vascular permeability factor activity in guinea pig skin, and lethal activity for mice . The specific activity of toxin preparations from cultures of a virulent strain of the bacterium was ca . 25-fold more than that of toxin preparations obtained from cultures of a weakly virulent strain . The four toxic activities were inseparable by gel filtration with Sephadex G-100; however, two components, which had markedly different elution behavior but which possessed the four activities mentioned above, were obtained . The major (ca . 88% of the recovered activity) and minor components had apparent molecular weights of ca . 38,500 and greater than 150,000, respectively. J Clin Invest, 1981 Aug, 68(2), 321 - 8 Properties of human asialo-factor VIII . A ristocetin-independent platelet-aggregating agent; De Marco L et al.; Human Factor VIII desialylated by treatment with Vibrio cholerae neuraminidase (ASVIII) aggregated human platelets in the absence of ristocetin in platelet-rich plasma and, to a lesser extent, in washed platelet suspensions . Aggregation is accompanied by thromboxane formation and is completely inhibited by EDTA . Aspirin blocks the second phase of aggregation and abolishes thromboxane production . Subaggregating doses of ASVIII and of either ADP, epinephrine, or collagen produce prompt and complete platelet aggregation . Bernard-Soulier syndrome platelets either did not aggregate with ASVIII (Two cases) or showed markedly decreased aggregation (one cases) . Factor VIII complex was prepared from the plasma of two patients with variant von Willebrand's disease (sialic acid content 142 and 75 nmol/mg, respectively); neither protein generated platelet-aggregating activity upon desialylation . {3H}ASVIII binds rapidly to platelets and 37 degrees C, while tritiated, fully sialylated factor VIII binds to a negligible extent . As little as 1--2 micrograms ASVIII bound/10(9) platelets is capable of inducing platelet aggregation . ASVIII may be a useful tool for investigating platelet-Factor VIII interactions in the absence of ristocetin . Furthermore, desialylated Factor VIII might play a physiologic role in Factor VIII-mediated platelet reactions in vivo. Nature, 1981 Jul 30, 292(5822), 413 - 17 Actions of cholera toxin and the prevention and treatment of cholera; Holmgren J; The drastic intestinal secretion of fluid and electrolytes that is characteristic of cholera is the result of reasonably well understood cellular and biochemical actions of the toxin secreted by Vibrio cholerae . Based on this understanding it is possible to devise new techniques for the treatment and prophylaxis of cholera to complement those based on fluid replacement therapy and sanitation. Biochemistry, 1981 Jul 7, 20(14), 4198 - 203 Potassium ion is required for the generation of pH-dependent membrane potential and delta pH by the marine bacterium Vibrio alginolyticus; Tokuda H et al.; The electrochemical potential gradient of protons in the marine bacterium Vibrio alginolyticus was measured as a function of external pH . In K+-containing cells, the membrane potential (delta psi) and delta pH vary with external pH as reported in other bacteria . On the other hand, K+-depleted cells show little pH dependence in the magnitude of delta psi from pH 6.0 to 8.5 . The cytoplasmic pH in these cells varies depending on external pH, resulting in the generation of a small delta pH at acidic pH . Addition of K+ to K+-depleted cells leads to partial dissipation of delta psi and concomitant generation of delta pH . Strikingly, this effect of K+ is dependent on external pH . Collapse of delta psi and generation of delta pH by the addition of K+ decrease with increasing external pH . Thus, the delta psi and delta pH obtained after addition of K+ are essentially the same as those determined in K+-containing cells, and cytoplasmic pH becomes less dependent on external pH . The results suggest that the variation of delta psi and delta pH with external pH is controlled by K+ transport. J Gen Microbiol, 1981 Jul, 125(Pt 1), 167 - 72 Adhesion of vibrios and aeromonads to isolated rabbit brush borders; Levett PN et al.; Isolated rabbit brush borders were used to investigate the adhesive properties of clinical and environmental isolates of non-O1 Vibrio cholerae and clinical isolates of Vibrio parahaemolyticus and Aeromonas hydrophila . A minority of the isolates were found to adhere to brush borders . The adhesion of these isolates was affected by a number of factors, including the bacteria:brush border ratio, incubation time, culture medium and growth temperature . Adhesion was inhibited by L-fucose but was independent on calcium ion concentration. Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Bacteriol Virusol Parazitol Epidemiol, 1981 Jul-Sep, 26(3), 145 - 54 {Current status of diarrhea caused by or associated with cholera vibrions of the non 0:1 group}; Ciufecu C et al.; Known until not long age under the generic name of non-agglutinable vibrios (NAG), cholera vibrios group non 0:1 represent a group as yet insufficiently known in spite of the marked progress made during the last 10 - 15 years in this field . Wide spread throughout the world vibrios group non 0:1 have been isolated from surface waters, from the intestines of fish, birds and mammals (from humans in case of enteritis, systemic infections, healthy subjects) . The medical interest stands in the participation of these vibrios in the acute diarrheic syndrome and as aetiologic agents . The authors review the data concerning the symptomatology, pathogenic mechanisms, epidemiology and bacteriologic diagnosis. Zh Mikrobiol Epidemiol Immunobiol, 1981 Jul, (7), 22 - 7 {Genetic control of toxin formation by Vibrio cholerae . I . Mobilization of the tox gene of Vibrio cholerae by plasmid RP::Mu ctsS62}; Skavronskaia AG et al.; The possibility of the mobilization of V . cholerae genes responsible for toxin formation (tox gene) and serological specificity (oag-I gene, serotype Inaba) by the RP4::Mu cts62 plasmid was established . The cotransfer of tox and oag-I genes with his and ilv genes respectively by means of the RP4::Mu cts62 plasmid was revealed . The frequency of the cotransfer of tox his and oag-I ilv observed in our experiments corresponded to the known frequency of the cotransfer of these genes observed in crosses with V . cholerae P+-strains . In recombination clones selected according to chromosomal amino acid markers (His+, Ilv+), both carrying and not carrying tox and oag-I genes, no markers of the RP4 plasmid and phage Mu cts62 were detected . The only exception was one His+-recombinant clone . It possessed thermosensitivity (the marker of phage Mu cts62), not linked, however, with sensitivity to phage thermoinduction . Similar clones were obtained when the plasmid was eliminated from V . cholerae strains containing the RP4::Mu cts62 plasmid and having the phenotype corresponding to this plasmid. Infect Immun, 1981 Jul, 33(1), 136 - 41 Nonimmunoglobulin fraction of human milk inhibits bacterial adhesion (hemagglutination) and enterotoxin binding of Escherichia coli and Vibrio cholerae; Holmgren J et al.; Human milk and colostrum samples were divided into an immunoglobulin and a nonimmunoglobulin fraction by immunosorbent chromatography . The ability of these fractions to inhibit bacterial cell adhesion and enterotoxin receptor binding of Vibrio cholerae and various Escherichia coli isolates was then tested by in vitro assays . The strongest effect was generally seen with the nonimmunoglobulin fractions, which were shown to significantly inhibit E . coli cell adhesion (hemagglutination) mediated by CFA/I, CFA/II, or K88 fimbriae (but not type 1 pili) and V . cholerae hemagglutination, as well as the binding of cholera toxin and E . coli heat-labile enterotoxin to GM1 ganglioside . Also, the immunoglobulin fractions had significant inhibitory activity in some of these systems . The results are interpreted to suggest that human milk and colostrum may contain secreted structure analogs of the cell receptors for some bacterial adhesions and enterotoxins; this might contribute to the protective effect of milk against enteric infections. J Bacteriol, 1981 Jul, 147(1), 36 - 45 Evolution of alkaline phosphatase in marine species of Vibrio; Woolkalis MJ et al.; The evolution of alkaline phosphatase was studied in marine species of Vibrio . Two antisera prepared against purified alkaline phosphatases from Vibrio splendidus and Vibrio harveyi were used to estimate the amino acid sequence divergence of this enzyme in 51 strains belonging to nine species . The methods used were the quantitative microcomplement fixation technique and the Ouchterlony double-diffusion procedure . There was a high degree of congruence between the measurement of the amino acid sequence divergence of alkaline phosphatase and the percentage of deoxyribonucleic acid homology of the different organisms relative to both reference strains (correlation coefficient of -0.89) as well as between the amino acid sequence divergence of alkaline phosphatase and superoxide dismutase (correlation coefficient of 0.92) relative to V . splendidus . These findings supported the view that the evolution of marine species of Vibrio is primarily vertical and that horizontal evolution (involving genetic exchange between species), if significant, is restricted to a minor fraction of the bacterial genome. Biken J, 1981 Jun, 24(1-2), 75 - 80 Presence and location of an O-acetyl group in O4-antigen of Vibrio parahaemolyticus; Tanaka S et al.; A polysaccharide moiety (PS) released from O4-LPS of Vibrio parahaemolyticus was purified . The constituent sugars of the purified PS were glucose, galactose, arabinose, fucose, heptose, N-acetyl galactosamine and an unknown N-acetyl amino sugar . The unknown amino sugar was deduced to be a terminal deoxy amino sugar from its NMR spectrum . PNMR spectroscopy at 200 MHz, periodate oxidation and treatment with alpha-L-fucosidase of the purified PS sample before and after alcoholic sodium methoxide-treatment suggested that PS contained an O-acetyl and alpha-linked non-reducing terminal L-fucose and that the O-acetyl group was located at the C-2 or C-4 position of the non-reducing terminal fucose . The contribution of the O-acetyl group located in L-fucose to the O4-antigenic determinant seemed to be small, judging from immunochemical experiments. Biull Eksp Biol Med, 1981 Jun, 91(6), 759 - 62 {Ultrastructural organization of tuft cells of the small intestinal epithelium}; Zufarov KA et al.; Electron microscopy was used to described a special cell type seen in the small intestinal epithelium of rats (conventional, sterile, monocontaminated with El Tor cholera vibrios, and three-day-old) . Ultrastructurally the cells in question appeared to be analogous to "brush" alveolocytes and tuft cells of the mucous membrane of the bronchi, stomach, gallbladder and eyes . It is suggested that these cells may be attributed to receptor cells performing resorptive and secretory functions. J Infect Dis, 1981 Jun, 143(6), 818 - 20 Duration of infection-derived immunity to cholera; Levine MM et al.; Four volunteers were rechallenged with Vibrio cholerae (10(6) classical Ogawa 395 organisms) 33-36 months after their initial induced cholera infection; none of the four veterans and four of five control volunteers developed diarrhea (P = 0.04) . All control subjects, but only one veteran, had positive coprocultures . Three of the four veterans had significant levels of serum IgG antitoxin before challenge, but none had measurable intestinal levels of secretory IgA antitoxin . Significant rises in levels of serum vibriocidal and antitoxic antibody occurred in all control subjects and in two veterans, who also manifested rises in levels of intestinal secretory IgA antitoxin . The impressive duration of infection-derived immunity suggests that the most promising approach to development of cholera vaccines may be to mimic natural immunity with orally administered, attenuated strains of V . cholerae. Infect Immun, 1981 Jun, 32(3), 1193 - 9 Edema and hemoconcentration in mice experimentally infected with Vibrio vulnificus; Bowdre JH et al.; Vibrio vulnificus (lactose-positive Vibrio), a recently recognized pathogenic marine species, produced extreme hemoconcentration and death within 3 to 6 h after subcutaneous or intraperitoneal injection of 10(8) viable cells into mice; hemotocrit values approached 70% (normal, 45%) . About 1 ml of edema fluid accumulated at the site of each subcutaneous injection, and locally increased vascular permeability was demonstrated by a skin bluing assay, using Evans blue dye . A corresponding fluid accumulation did not occur in the peritoneal cavity after an intraperitoneal injection . Filter-sterilized supernatants of cultures grown under a variety of conditions did not produce local edema or lethality, nor did whole Vibrio cells killed by a variety of methods or disrupted by sonic oscillation . Edema fluids collected from infected mice and sterilized by filtration had no effect when they were injected subcutaneously or intraperitoneally into mice . Inocula of 10(9) viable cells of V . vulnificus contained within a diffusion chamber implanted subcutaneously did not produce skin bluing, edema, or lethality; Vibrio cells remained viable and virulent within these chambers for at least 2 weeks . These experiments suggested that vascular permeability changes in V . vulnificus infections may not be attributable to a diffusible toxin and may require direct contact between host cells and viable Vibrio cells. J Bacteriol, 1981 Jun, 146(3), 1038 - 45 Luciferase inactivation in the luminous marine bacterium Vibrio harveyi; Reeve CA et al.; Luciferase was rapidly inactivated in stationary-phase cultures of the wild type of the luminous marine bacterium Vibrio harveyi, but was stable in stationary-phase cultures of mutants of V . harveyi that are nonluminous without exogenous aldehyde, termed the aldehyde-deficient mutants . The inactivation in the wild type was halted by cell lysis and was slowed or stopped by O2 deprivation or by addition of KCN and NaF or of chloramphenicol . If KCN and NaF or chloramphenicol were added to a culture before the onset of luciferase inactivation, then luciferase inactivation did not occur . However, if these inhibitors were added after the onset of luciferase inactivation, then luciferase inactivation continued for about 2 to 3 h before the inactivation process stopped . The onset of luciferase inactivation in early stationary-phase cultures of wild-type cell coincided with a slight drop in the intracellular adenosine 5'-triphosphate (ATP) level from a relatively constant log-phase value of 20 pmol of ATP per microgram of soluble cell protein . Addition of KCN and NaF to a culture shortly after this drop in ATP caused a rapid decrease in the ATP level to about 4 pmol of ATP per microgram whereas chloramphenicol added at this same time caused a transient increase in ATP level to about 25 pmol/microgram . The aldehyde-deficient mutant (M17) showed a relatively constant log-phase ATP level identical with that of the wild-type cells, but rather than decreasing in early stationary phase, the ATP level increased to a value twice that in log-phase cells . We suggest that the inactivation of luciferase is dependent on the synthesis of some factor which is produced during stationary phase and is itself unstable, and whose synthesis is blocked by chloramphenicol or cyanide plus fluoride. Cancer Res, 1981 Jun, 41(6), 2262 - 6 A correlation between cell surface sialyltransferase, sialic acid, and glycosidase activities and the implantability of B16 murine melanoma; Dobrossy L et al.; A murine melanoma variant (B16-F10ir6), resistant to lymphocytic cytolysis, has been shown previously to produce lower numbers of tumor nodules in the lung of C57BL/6J mice following i.v . inoculations . These differences found in tumor implantation and lymphocyte recognition may be due to changes in surface properties of this cell line . Therefore, membrane-bound sialic acid (released by Vibrio cholerae neuraminidase treatment), ectosialyltransferase activity, and total cellular glycosidase levels were measured in this cell line and compared with levels in its parent melanoma tumor cell line, B16-F10, which was selected for its enhanced ability to form tumor nodules . The results of these studies indicate a correlation between the degree of lung implantation and the amount of tumor cell sialic acid accessible to neuraminidase cleavage, tumor cell surface sialyltransferase activity, and several cellular glycosidase activities . These results are consistent with the idea that membrane structural changes in the glycocalyx may account for the ability of a tumor cell to implant and metastasize. Infect Immun, 1981 Jun, 32(3), 1132 - 8 Transposon-facilitated recombination in classical biotypes of Vibrio cholerae; Sublett RD et al.; Transposon-facilitated recombination (Tfr) donors of classical Vibrio cholerae strain 162 were constructed by introducing the ampicillin transposon Tn1 into the P conjugative plasmid and the bacterial chromosome . The improved donors mediated high-frequency, polarized transfer of chromosomal genes from origins to confirm the gene orders of the previous classical strain 162 genetic map and to establish its circularity . Significant transfer of linked genes from E1 Tor Tfr donors to classical recipients was demonstrated, and other evidence for genetic relatedness of these two V . cholerae biotypes is discussed. Ann Hum Biol, 1981 May-Jun, 8(3), 289 - 91 ABO blood group distributions in diarrhoea cases including cholera in Calcutta; Sircar BK et al.; A double blind study on determination of ABO blood groups of 210 cholera patients, 44 diarrhoea cases due to Vibrio parahaemolyticus, and 148 diarrhoea cases from whom no vibrios could be isolated (control group), was conducted in Calcutta . A statistic