Eur J Biochem, 1984 Feb 15, 139(1), 195 - 200 Structure of the O-chain of the phenol-phase soluble cellular lipopolysaccharide of Yersinia enterocolitica serotype O:9; Caroff M et al.; The phenol-phase soluble cellular lipopolysaccharide isolated by the phenol/water extraction method from Yersinia enterocolitica serotype O:9 cells was shown by hydrolytic, periodate oxidation, methylation and nuclear magnetic resonance studies to be an S-type lipopolysaccharide with a linear O-antigenic polysaccharide of 1,2-linked 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl units . The serological cross-reactivity between Y . enterocolitica serotype O:9 and the lipopolysaccharides of Vibrio cholerae and Brucella species can now be related to the presence of N-acylated 4-amino-4,6-dideoxy-alpha-D-mannopyranosyl residues in their respective O-antigenic chains. Lancet, 1984 Feb 11, 1(8372), 325 - 8 Foodborne transmission of cholera in Micronesian households; Holmberg SD et al.; During a cholera epidemic in Truk (Micronesia), in a survey of 1 village, transmission of Vibrio cholerae O1 in the 28 households with illness appeared to be through food contaminated in the home . Households in which the index case was a foodhandler had significantly higher attack rates than households in which the index case was not a foodhandler . Members of households with illness were significantly more likely to become ill if they had eaten food prepared by a foodhandler who had recently been ill . A matched-pair case-control study on several adjacent islands confirmed this relation . Those who had tended an ill person were not at increased risk of cholera in either study . This study suggests that in this outbreak, foodborne transmission of V cholerae O1 was more important than contact spread. Infect Immun, 1984 Feb, 43(2), 515 - 22 Evaluation in humans of attenuated Vibrio cholerae El Tor Ogawa strain Texas Star-SR as a live oral vaccine; Levine MM et al.; Texas Star-SR, an A- B+ mutant derived by nitrosoguanidine treatment from Vibrio cholerae El Tor Ogawa strain 3083, was fed to 68 volunteers as an oral vaccine in doses of 10(5) to 5 X 10(10) organisms with NaHCO3 . Sixteen (24%) vaccinees experienced some loose stools (unrelated to vaccine dose), but in only one did the total stool volume exceed 1.0 liter . The vaccine strain was cultured from duodenal fluid of 35 of 46 (76%) persons who ingested doses of 10(8) organisms or greater . No A+ B+ toxinogenic revertants were found among 456 clinical isolates tested . Sixty-three vaccinees (93%) manifested seroconversions of vibriocidal antibody, whereas only 20 (29%) had significant rises in serum antitoxin titers . Paired intestinal fluids from 41 volunteers showed significant rises of secretory immunoglobulin A against lipopolysaccharide (29%), Ogawa outer membrane preparation (29%), and toxin (12%) antigens . In challenge studies with pathogenic V . cholerae El Tor Ogawa and El Tor Inaba, the attack rate in vaccinees (7 of 25) was significantly lower than in controls (18 of 25) (vaccine efficacy, 61%); furthermore, the diarrheal stool volume in vaccinees was significantly less than that in controls (P less than 0.01) . Texas Star-SR served as a prototype to investigate the concept of immunoprophylaxis by means of attenuated strains as oral vaccines . These observations provide an invaluable background for planning future studies with newly developed attenuated strains prepared by recombinant DNA techniques. J Hyg (Lond), 1984 Feb, 92(1), 59 - 65 Transferable plasmid-mediated drug resistance among non-O1 Vibrio cholerae and rough strains of Vibrio cholerae from Tamilnadu, India; Sundaram S et al.; A total of 289 non-O1 Vibrio cholerae (NVC) strains and 20 rough V . cholerae (RVC) strains isolated in an endemic area were tested for antibiotic resistance and for transferable R-plasmids . Twenty three per cent of NVC and 40% of the RVC isolates were found to be resistant to one or more drugs . Eight NVC and four RVC strains possessed multiple drug resistance, varying from four to eight drugs . The common spectrum found in NVC isolates were chloramphenicol and streptomycin (CS) or chloramphenicol, streptomycin, tetracycline and ampicillin (CSTA) . Resistance to sulphamethoxazole (Su) and to trimethoprim (Tm) was encountered infrequently . In RVC isolates in addition CSTASuTm determinants, resistance markers to aminoglycosides kanamycin, gentamicin and neomycin were also found . Eighteen of the 27 V . cholerae strains with two or more resistance determinants transferred them en bloc to Escherichia coli K12 . The level of resistance in the recipient strain was equal to or greater than that of the donor vibrio strains . Most of the strains possessing solitary resistance markers were unable to transfer them . beta-lactamase production could be demonstrated in 92.8% of the ampicillin resistant strains . None of the strains was resistant to nalidixic acid or furazolidone . The results emphasize the importance of antimicrobic susceptibility determination of V . cholerae isolates, regardless of the serotypes, before commencing chemotherapy. Zh Mikrobiol Epidemiol Immunobiol, 1984 Feb, (2), 97 - 9 {Determination of cholera enterotoxin in supernatants of homogenates from the wall of the small intestine in rabbits in the passive immune hemolysis reaction}; Gailonskaia IN et al.; The article presents the data indicating that suckling rabbits can be used as a model, specific only for enterotoxin-producing Vibrio eltor strains . The signs characteristic of low virulence may appear as the consequence of the action of other products resulting from the vital activity of vibrios . No strains capable of producing toxin only in vivo or in vitro have been detected . The quantitative correlation of the synthesis of enterotoxin in the body of experimental animals and in vitro has been established . The passive immune hemolysis test can be used as an objective method suitable for the determination of the choleragenic potency of vibrios. Infect Immun, 1984 Feb, 43(2), 765 - 7 Lack of homology between the iron transport regions of two virulence-linked bacterial plasmids; Walter MA et al.; Two plasmids involved in bacterial virulence, the Escherichia coli plasmid pColV-K30 and the Vibrio anguillarum plasmid pJM1, have been studied with respect to the iron sequestering systems mediated by these two plasmids . Bioassay results show that the two systems are not related functionally because specific iron uptake-deficient mutants in each system cannot be cross-fed by the heterologous bacteria using culture supernatants from iron-proficient strains containing wild-type plasmids . DNA hybridization studies show an extensive lack of homology between regions involved in iron sequestration in both plasmids. Acta Pathol Microbiol Immunol Scand {B}, 1984 Feb, 92(1), 53 - 60 Intestinal adaptation to cyclic AMP-mediated hypersecretion induced by the heat-labile enterotoxin of Vibrio cholerae and Escherichia coli; Lonnroth I et al.; Adaptation to cholera toxin (CT) and the heat-labile enterotoxin (LT) from E . coli is studied in vivo in the rat small intestine . Repeated peroral pretreatment with CT or LT induces protracted inhibition of the intestinal fluid response to these toxins . The CT-induced mucus release from intestinal goblet cells is not influenced by CT pretreatment and the binding of CT to the epithelium remains intact . However, the adenylate cyclase activity, which mediates CT and LT action, is repressed--as judged from the response of this enzyme to both CT, LT and prostaglandin E1 . The results suggests that protection against CT and LT acquired in the gut is achieved by desensitization of the adenylate cyclase system. J Antimicrob Chemother, 1984 Feb, 13(2), 171 - 5 Pivmecillinam, co-trimoxazole and oral mecillinam in gastroenteritis due to Vibrio spp; Uylangco C et al.; The comparative efficacy of antibacterial therapy with pivmecillinam or cotrimoxazole and general supportive care only was studied in patients with severe bacterial gastroenteritis . Overall, treatment with antibiotics proved significantly superior to rehydration alone in 42 children . Active therapy also had a statistically beneficial effect in children infected with Vibrio cholerae and V . parahaemolyticus . Pivmecillinam and co-trimoxazole were equally effective . Pivmecillinam and oral mecillinam appeared to be of equal value in a further 22 adults infected by Vibrio spp . No side-effects were recorded in any of the subjects treated . Further investigations with pivmecillinam and oral mecillinam are advocated. J Clin Microbiol, 1984 Feb, 19(2), 296 - 7 Isolation of nontoxigenic Vibrio cholerae O group 1 from a patient with severe gastrointestinal disease; Morris JG Jr et al.; A nontoxigenic strain of Vibrio cholerae O group 1 was isolated in Florida from the stool of a patient with severe diarrhea . The strain had the same hemolytic and unique phage-sensitivity pattern as all toxigenic isolates from recent cases of cholera in Texas and Louisiana . Identical strains were transiently isolated from sewerage systems in two other Florida communities, suggesting that multiple human infections had occurred . This is the first indication that V . cholerae O1 strains which do not produce cholera toxin may be able to cause gastrointestinal disease in humans . The identification of these strains also raises questions about the relationship between toxigenic and nontoxigenic strains of V . cholerae O1 along the Gulf Coast of the United States. J Clin Microbiol, 1984 Feb, 19(2), 181 - 6 Production of Vibrio cholerae O1 and non-O1 typing sera in rabbits immunized with polysaccharide-protein carrier conjugates; Adams LB et al.; Two systems are currently used to serologically type Vibrio cholerae O1 and non-O1 isolates . Antiserovar-serotype serum in the Smith system is produced in rabbits immunized with live whole-cell vaccines, and that in the Sakazaki system is produced in rabbits immunized with heat-killed vaccines . In neither system is the serovar-serotype-specific antigen clearly defined . During the course of a serological survey, ca . 10% of more than 2,500 V . cholerae isolates examined agglutinated in the optimal dilutions of two, three, or four different anti-serovar sera prepared by the methods of Sakazaki . An occasional isolate agglutinated in both anti-O1 and non-O1 sera . Lipopolysaccharide was extracted from eight of these possible multiple serovars, coated onto rabbit erythrocytes, and retested in these same antisera by passive hemagglutination . With one exception the lipopolysaccharide-rabbit erythrocytes were now agglutinated in a single antiserum . Antipolysaccharide sera were produced in rabbits immunized with the polysaccharide moiety extracted from eight non-O1 and two O1 vaccine strains conjugated to bovine gamma globulin protein carrier . The antipolysaccharide sera showed passive hemagglutination titers versus lipopolysaccharide-rabbit erythrocytes comparable to those achieved in antisera from rabbits immunized with heat-killed whole-cell vaccines . In the slide agglutination test antipolysaccharide sera serologically discriminated between two O1 isolates that were previously agglutinated in both anti-O1 and anti-non-O1 whole-cell sera . It is recommended that serological types or varieties of V . cholerae non-O1 be based upon serologically recognizable differences in lipopolysaccharide-associated antigens as are antigens A, B, and C in the O1 group. Biochem Biophys Res Commun, 1984 Jan 13, 118(1), 82 - 9 Sialosylglobotetraosylceramide: a marker for amyotropic lateral sclerosis; Kundu SK et al.; Gangliosides of healthy and pathologic muscles (amyotropic lateral sclerosis and facio-scapulo-humeral muscular dystrophy) were studied . Total ganglioside content of the affected muscles was approximately 2 fold higher than the unaffected muscles . Our results showed that ALS muscle contained a ganglioside which was absent in the unaffected and FSH muscular dystrophic muscles . Based on the results of hydrolysis with Vibrio cholerae neuraminidase and subsequent reactivity of the asialo derivative towards anti-globotetraosylceramide, we propose that the ALS ganglioside is sialosylglobtetraosylceramide, NeuAc(alpha 2-3)Ga1NAc(beta 1-3)Ga1(alpha 1-4)Ga1(beta 1-4)G1c-Cer. J Biol Chem, 1984 Jan 10, 259(1), 383 - 5 Vibriobactin, a siderophore from Vibrio cholerae; Griffiths GL et al.; A novel siderophore (microbial iron transport compound) has been isolated from low iron cultures of Vibrio cholerae . Belonging to the catecholamide family of chelators, it has been shown to contain three residues of 2,3-dihydroxybenzoic acid and two residues of threonine . Both threonine moieties are present in the form of oxazoline rings . Furthermore, the polyamine backbone of the molecule was proved to be not spermidine, but the rare N-(3-aminopropyl)-1,3-diaminopropane, norspermidine . The structure of the new siderophore has been determined to be N-{3-(2,3-dihydroxybenzamido)propyl}-1, 3-bis{2,3-dihydroxyphenyl)-trans-5-methyl-2-oxazoline-4-carboxamido}prop ane . The compound has been given the trivial name vibriobactin . Mutants defective in the synthesis and utilization of vibriobactin were isolated . In an iron-limited environment V . cholerae was found to respond more strongly to vibriobactin, agrobactin, and ferrichrome than to enterobactin. Med J Aust, 1984 Jan 7, 140(1), 22 - 3 Vibrio vulnificus septicaemia; Brady LM et al.; A patient, who had been receiving treatment for refractory anaemia with excess of myeloblasts for three years, developed septic shock due to Vibrio vulnificus septicaemia . The infection was believed to have been introduced with the ingestion of raw oysters . This organism is comparatively rare, but it can cause serious infection, and is known to be capable of very rapid tissue invasion. Int Arch Allergy Appl Immunol, 1984, 75(1), 38 - 43 Gut mucosal, salivary and serum antitoxic and antibacterial antibody responses in Swedes after oral immunization with B subunit-whole cell cholera vaccine; Jertborn M et al.; Gut mucosal, salivary and serum antibody responses to a new oral cholera vaccine, consisting of B subunit and whole cell vaccine (WCV), were studied in Swedish volunteers . A single immunization with a 0.5 mg dose of B subunit together with WCV (5 X 10(10) killed cholera vibrios) induced a local intestinal immunoglobulin A (IgA) antitoxin response in 4/6 (67%) vaccine recipients as evident from specific antibody titre rises in intestinal lavage fluid . A second administration of vaccine did not further enhance the intestinal immune response beyond the peak level induced by the initial immunization . Different doses of B subunit (2.5 and 0.5 mg) given together with 5 X 10(10) killed vibrios (WCV) induced antitoxin antibody responses in serum in about the same frequency, 10/13 (77%) responders versus 13/14 (93%), as well as in saliva, 7/13 (54%) versus 9/14 (64%), and a single immunization was almost as efficient as two vaccine administrations . Single or repeated oral vaccination only irregularly resulted in modest antibacterial titre rises in serum (9/27 = 33%) or saliva (12/27 = 44%), but stimulated a significant mucosal antibacterial response in intestine of 5/6 (83%) examined volunteers. Microbiol Immunol, 1984, 28(3), 327 - 37 Isolation and characterization of a filamentous phage, Vf33, specific for Vibrio parahaemolyticus; Taniguchi H et al.; Phage Vf33, a filamentous phage about 1,400 nm long and 7 nm wide, specific for Vibrio parahaemolyticus, was isolated and characterized . The buoyant density of Vf33 in CsCl was 1.292 g/cm3 . As with other filamentous phages, the lytic activity of Vf33 was resistant to heating below 80 C and to treatment with diethylether, acetone or methanol but sensitive to chloroform . The nucleic acid of this phage is single-stranded circular DNA 8.4 kb in size . The viral genome was converted to a double-stranded replicative form in the host-cell . Among the strains tested, only V . parahaemolyticus strains possessing K38 antigen was sensitive to the phage. Microbiol Immunol, 1984, 28(3), 311 - 25 Ecology of non-O 1 Vibrio cholerae in Toyama Prefecture; Kodama H et al.; The ecology of non-O 1 Vibrio cholerae and Vibrio mimicus as causes of cholera-like diarrhea or seafood-associated gastroenteritis has been investigated in Toyama Prefecture since 1980 . The relationship between biological or serological characteristics of the isolates and their enteropathogenicity is discussed . Overall isolation rates from river water, sea water, and fish were 24.0, 59.5, and 33.7%, respectively, the isolation frequency being, in general, extremely high in the summer season, although the organisms were detected all year around in the case of sea water . Most isolates from river water were unable to grow on plates of TCBS agar to which colistin was added at a concentration of 1 microgram/ml (CL-TCBS) . These strains quickly fermented cellobiose . O-51 and O-70 were the two most frequently detected serogroups among them and they did not show enteropathogenicity in the rabbit ileal loop ( RIL ) test . On the other hand, almost all isolates from sea water and fish as well as those from human diarrhea cases were able to grow on CL-TCBS, but were unable to ferment cellobiose quickly . O-36, O-10, O-6, O-8, O-39, and O-26 were the dominant serogroups of these isolates, and some of them showed enteropathogenicity in the RIL test . Six out of 98 isolates from river water, 14 out of 116 from sea water, and 19 out of 112 from fish were classified as Vibrio mimicus . All of these strains were able to grow on CL-TCBS and quickly fermented mannose but not cellobiose . O-41 was the most common serogroup among them and some of these strains showed enteropathogenicity in the RIL test . Production of a cholera-like enterotoxin among the isolates in Toyama Prefecture, if any, seemed to be poor. Microbiol Immunol, 1984, 28(1), 1 - 9 Drug resistance and R plasmids in Vibrio anguillarum isolated in cultured ayu (Plecoglossus altivelis); Aoki T et al.; Two hundred twenty-six strains of Vibrio anguillarum collected from cultured ayu ( Plecoglossus altivelis ) between 1978 and 1980 were studied for their sensitivities to 10 chemotherapeutics . In order to determine whether the drug-resistant strains possessed transferable R plasmids, they were conjugated with Escherichia coli . Almost all the strains isolated during the 3 years showed resistance to nalidixic acid (NA) and/or furazolidone (NF) . NA and NF resistance were not transferred to Escherichia coli from any of the strains . Chloramphenicol-resistant strains were isolated in every year and almost all of them carried transferable R plasmids . Only one strain with tetracycline resistance was found among the strains tested . Strains resistant to sulfonamides, streptomycin, ampicillin (ABP), and trimethoprim (TMP) increased rapidly in 1980, and a large number of them carried transferable R plasmids . Transferable R plasmids encoded with resistance to ABP and TMP were detected for the first time in V . anguillarum strains . The R plasmids detected in the strains isolated in 1980 were classified into incompatibility groups E, A, and an untypable group . The R plasmid DNAs were cleaved by EcoRI to yield 11 to 13 fragments . The estimated molecular weights of the R plasmids from the five strains ranged from 97 to 104 M daltons. J Clin Microbiol, 1984 Jan, 19(1), 87 - 8 Isolation of mucoid Vibrio parahaemolyticus strains; Lam S et al.; Mucoid strains of Vibrio parahaemolyticus were isolated from the stools of two asymptomatic carriers and a patient with gastroenteritis . The strains demonstrated biochemical reactions and antibiotic susceptibility typical of nonmucoid strains of V . parahaemolyticus isolated locally . The slime substance was typed by coagglutination and was antigenically similar to the capsular antigen of the same strain . Three different serotypes (O10:K24, O5:K17, and O5:K15) were involved. J Bacteriol, 1984 Jan, 157(1), 253 - 61 Genetic mapping of Vibrio cholerae enterotoxin structural genes; Sporecke I et al.; The structural genes which constitute the cholera toxin operon, ctxAB, were genetically mapped in the Vibrio cholerae El Tor strain RV79 . This strain of V . cholerae contains two copies of the ctx operon located on a 7-kilobase-pair tandemly duplicated region . We began by isolating a vibriophage VcA1 insertion mutation in one of the two ctxA genes located in this region . The mutant carrying this ctxA::VcA1 insertion, DC24, was converted to a VcA1-facilitated donor by introduction of the conjugal plasmid pSJ15, which carries an inserted copy of a defective VcA1-like prophage . The donor characteristics of DC24(pSJ15) indicated that the ctxA::VcA1 insertion mutation was near the trp region of the V . cholerae chromosome . Subsequent RV79 three-factor crosses were performed between VcA1-facilitated donors and recipient strains carrying one of two structural gene mutations in ctx, either delta ctxA23P Kmr or delta ctx-7922 . The former was constructed by an in vivo marker exchange procedure and could be scored either by its kanamycin resistance phenotype or by its lack of DNA sequences homologous to the ctxA region . The delta ctx-7922 mutation is a total deletion of both ctx copies of strain RV79 . The three-factor cross data strongly suggest that the two ctx loci of RV79 map between the nal and his genes of V . cholerae in the trp nal his linkage group . Physical analysis and heterologous crosses between an RV79 El Tor donor and a 569B classical recipient indicates that one of the two 569B ctx operon copies maps in the same region as the RV79 ctx loci (i.e., linked to nal) . Together with previously published observations, these data show that the ctx structural genes are not closely linked to other genes known to affect toxin production in V . cholerae. Microbiol Immunol, 1984, 28(7), 735 - 45 Studies on the enteropathogenic mechanism of non-O 1 Vibrio cholerae isolated from the environment and fish in Toyama Prefecture; Gyobu Y et al.; Enteropathogenic mechanisms of non-O 1 Vibrio cholerae were investigated using strains from the environment and those from fish in Toyama Prefecture . None of the 93 non-O 1 V . cholerae strains produced a detectable level of choleratoxin-like-enterotoxin (CT-like-enterotoxin) in Syncase medium, while 23 strains showed a distinct fluid accumulation in the rabbit ileal loop test (RIL) . These RIL-positive strains neither produced CT-like-enterotoxin in vitro in the other four kinds of media which are considered suitable for CT production, nor in vivo in the ligated ileal loop . Approximately one-third of RIL-positive strains produced a fluid accumulating factor (FAF) which was not neutralized with anti-CT serum . FAF of a representative strain (Strain 79-9-2) was inactivated by heating at 100 C for 10 min, and has a molecular weight within the range of 50,000 to 100,000 daltons . Most accumulated fluids in RIL after inoculation with whole cultures of RIL-positive strains contained both hemolytic and cytotoxic principles . Desquamation of epithelial cells, inflammatory edema, neutrophile infiltration, loss of goblet cells and frequent hemorrhages were observed in sections of ligated ileal loop inoculated with whole cultures or concentrated culture filtrates of CT-like-enterotoxin-negative but RIL-positive strains . In contrast, neither desquamation of epithelial cells nor hemorrhage was observed in sections after inoculation with those of a CT-like-enterotoxin positive strain (Strain E 8498) . These results indicated that most RIL-positive non-O 1 V . cholerae strains from the environment and fish isolated in Toyama Prefecture produce little CT-like-enterotoxin, but some of them produce FAF with cytotoxic activities. Biomed Biochim Acta, 1984, 43(6), 711 - 7 Interaction between rat peritoneal macrophages and sialidase-treated erythrocytes: biochemical and morphological studies; Schauer R et al.; Rat peritoneal macrophages bind and phagocytose homologous, sialidase-treated erythrocytes at a rate depending on the number of red cells and the amount of sialic acids released . Vibrio cholerae sialidase only partially (75%) removes the sialic acid residues from rat erythrocytes, whereas with Arthrobacter ureafaciens sialidase complete desialylation is possible . Analysis of the sialic acids by capillary gas-liquid chromatography combined with mass spectrometry (GLC-MS) revealed the occurrence of N-acetylneuraminic acid (Neu5Ac), N-acetyl-9-O-acetylneuraminic acid (Neu5,9Ac2), N-acetyl-7,9-di-O-acetylneuraminic acid (Neu5,7,9Ac3), N-acetyl-9-O-lactylneuraminic acid (Neu5Ac9Lt) and N-glycolyneuraminic acid (Neu5Gc) . Native rat serum enhances binding and phagocytosis, as has been observed by radioactive measurements and studies in a micro-scale by light and electron microscopy . The morphological experiments showed that maximum binding of sialidase-treated erythrocytes to macrophages occurs after 15-30 min, while for maximum phagocytosis at least 60 min are necessary . Striking alterations of the shape of erythrocytes during their interaction with macrophages were observed. Intervirology, 1984, 22(2), 61 - 71 Classification of Vibrio bacteriophages; Ackermann HW et al.; 85 Vibrio phages, 84 of them tailed and 1 filamentous, were surveyed . The tailed phages belonged to six basic morphotypes and to the Myoviridae, Siphoviridae, or Podoviridae families . 63 phages were classified into 18 species . The filamentous phage is a member of the Inovirus genus of the Inoviridae family . Vibrio phages are very heterogenous and include some morphologically interesting viruses . Several Vibrio phages closely resemble phages of other gram-negative bacteria, possibly indicating phylogenetic relationships between their hosts. Virus Res, 1984, 1(5), 401 - 14 Saturable attachment sites for type 3 mammalian reovirus on murine L cells and human HeLa cells; Gentsch JR et al.; Attachment of {35S}methionine-labelled mammalian type 3 reovirus to murine L cells and human HeLa cells was studied under equilibrium conditions . Cellular attachment sites could be completely saturated with 35S-labelled reovirus, indicating that specific attachment sites for reovirus are present on the surface of these cells . We calculated that L cells possess about 86000-105000 attachment sites per cell while HeLa cells possess about 126000-147000 sites per cell for type 3 reovirus . Unlabelled reovirus was highly efficient in competing for attachment by 35S-labelled reovirus to the saturable attachment sites of both L and HeLa cells, further indicating the specificity of the interaction . We also found that unlabelled reovirus competed equally well for both binding and internalization of 35S-labelled reovirus into murine L cells, suggesting that the L cell attachment site may serve as a virus entry site . Phospholipase digestion of L cells had no effect on subsequent reovirus attachment, while treatment of L cells with moderate concentrations of bromelain (but not trypsin, proteinase K or pronase) and Vibrio cholerae neuraminidase reproducibly decreased subsequent reovirus attachment . These results and those of others (Epstein et al., 1984, Virology 133, 46-55) suggest that mammalian reoviruses attach to specific cell surface receptors on at least two species of mammalian cells to initiate the infectious cycle. Microbiologica, 1984 Jan, 7(1), 11 - 9 B-galactosidase activity in different toxigenic and nontoxigenic strains of Vibrio cholerae: effect of glucose; Chakrabarti MK et al.; B-galactosidase activity in presence and absence of inducer lactose was examined in eight different strains of both toxigenic and nontoxigenic Vibrio cholerae (V . cholerae) . In lactose (0.25 gm%) containing cultures of toxigenic strains, B-galactosidase activity was higher in stationary culture than that of logarithmic . In nontoxigenic strains, B-galactosidase activity was only 1/6th of the toxigenic strains and remained unaltered throughout growth period . While wild strains (toxigenic V . cholerae) showed B-galactosidase activity at reduced rate when grown in the medium containing both lactose and glucose, the rate of activity of B-galactosidase in nontoxigenic strains remained unchanged whether the cells were grown in lactose or lactose plus glucose . The respressive effect of higher concentration of glucose could be reversed by the addition of exogenous cyclic AMP . Low level of B-galactosidase activity in nontoxigenic strains was due to low level of intracellular cyclic AMP, compared to toxigenic strain . Furthermore, induction of enzyme activity occurred in nontoxigenic strains of V . cholerae when cyclic AMP was added exogenously. J Hyg Epidemiol Microbiol Immunol, 1984, 28(2), 177 - 84 Investigation of populations heterogeneous according to O-antigen in Vibrio cholerae cultures; Andrusenko IT et al.; The O-antigenic composition of 36 cultures of Vibrio cholerae agglutinating simultaneously with 01 cholera sera and 0 sera to NAG vibrios of the Sakazaki collection was investigated . It has been established experimentally that under the effect of medium and environmental conditions such cultures dissociate to subcultures differing in their affiliation to different serological groups according to 0 antigen . The passage of these cultures in the organism of susceptible animals promotes preservation of 01-group clones whereas the passage in peptone water or prolonged storage under unfavourable conditions result in the predominance of clones of different serological affiliation . The proposition has been put forward that the observed vibrio cultures are genotypically capable of producing, besides the 01 group, a number of 0 antigens . Phenotypical manifestation of the antigenic structure in the respective individuals of the population depends on the conditions of the environment. Trans R Soc Trop Med Hyg, 1984, 78(6), 818 - 20 Role of rotavirus as the cause of acute paediatric diarrhoea in Calcutta; Saha MR et al.; Of the 245 acute paediatric diarrhoea cases admitted to the Infectious Diseases Hospital, Calcutta between July, 1979 and June, 1981, rotavirus was detected in the faeces of 55 (22.4%) patients either as the sole aetiological agent or in association with other enteropathogens . Children aged six months to two years were most commonly infected . The virus was detected throughout the year with higher incidence during the winter months of both years . The frequency of detection of major enteropathogens other than rotavirus was Vibrio cholerae biotype El Tor (31.0%), enteropathogenic Escherichia coli (8.2%) and enterotoxigenic E . coli (6.5%). Nature, 1983 Dec 8-14, 306(5943), 551 - 7 Cholera toxin genes: nucleotide sequence, deletion analysis and vaccine development; Mekalanos JJ et al.; Nucleotide sequence and deletion analysis have been used to identify the regulatory and coding sequences comprising the cholera toxin operon (ctx) . Incorporation of defined in vitro-generated ctx deletion mutations into Vibrio cholerae by in vivo genetic recombination produced strains which have practical value in cholera vaccine development. Aust J Exp Biol Med Sci, 1983 Dec, 61 ( Pt 6), 705 - 11 Modulation of antibody responses to Vibrio cholerae in mice by adoptive transfer of Peyer's patch lymphocytes from orally immunised donors; Ahmed A et al.; Peyer's patch lymphocytes from mice orally primed with V . cholerae injected into recipient mice together with V . cholerae led to a considerable suppression of IgM antibody-forming cells in the spleen by comparison with control mice receiving the same schedule but with Peyer's patch lymphocytes from unprimed mice . The effect on the splenic IgA responses of recipients was variable . Incomplete separation of the Peyer's patch cells showed that both T and B cell enriched fractions were active in this suppressive effect. Southeast Asian J Trop Med Public Health, 1983 Dec, 14(4), 548 - 58 Immunogenicity of soluble haemagglutinin-lipopolysaccharide complex of classical vibrio cholerae; Chaicumpa W et al.; Soluble haemagglutinin-lipopolysaccharide complexes were found to be good antigens since it elicited high levels of the antibodies in the intestine especially of the IgA class . These specific antibodies sustained for a long period of time at the significantly high levels (longer than 6 months) . The enteric memory primed by the antigens in the intestinal tract were longer than 3 months . Pools of intestinal fluids obtained from mice immunized with single dose of SH-LPS at 1 week, 1 month and 3 months after the antigen stimulation conferred protection against the homologous challenge . Better protection was found in the corresponding specimens collected from mice which received antigen booster at 3 months after the first stimulation . Multiple oral doses of the antigens (three doses at weekly intervals) did not have any advantage over the single dose immunization . The intestinal fluids obtained from the former group conferred similar degree of protection as those from the latter though the serum specimens offered higher PD50 . The protection against cholera does not correlate with the levels of vibriocidal antibody in the body fluids which seems to confirm the hypothesis that the mechanism of protection against cholera by intestinal antibody is by reducing or preventing the attachment of the vibrios to the epithelium either via agglutinating process or via masking of the vibrio adhesive sites. Appl Environ Microbiol, 1983 Dec, 46(6), 1438 - 41 Bacterial chemotaxis to effluent from a rum distillery in tropical near-shore coastal waters; Fuentes FA et al.; Pseudomonas aeruginosa and Vibrio cholerae showed a strong positive chemotactic response towards rum distillery wastewaters (mostos) and a high oxygen uptake rate in the presence of this complex substrate . Rum slops stimulated only motility in Aeromonas hydrophila and Escherichia coli . The A . hydrophila and E . coli isolates were unable to oxidize mostos significantly. J Infect Dis, 1983 Dec, 148(6), 998 - 1004 Phage types of Vibrio cholerae O1 biotype El Tor isolated from patients and family contacts in Bangladesh: epidemiologic implications; Glass RI et al.; The epidemiology of Vibrio cholerae O1 El Tor in rural Bangladesh was examined with a new phage-typing system for characterization of individual strains . During a two-year period, 537 strains were typed with a set of standard and experimental phages . Four major and many minor patterns were identified, some associated with discrete outbreaks of disease and others persisting for the entire period . One outbreak-related phage type was associated with a C plasmid bearing multiple drug resistance . The yearly cholera seasons began with the emergence of strains of many different phage types in many different locations; this pattern is consistent with an environmental reservoir for V cholerae . Each patient with cholera excreted only one phage type over time, whereas some family members were infected with strains of different phage types . Phage typing can be a useful marker with which the spread of cholera strains can be traced in endemic settings. Infect Immun, 1983 Dec, 42(3), 990 - 6 Vibrio cholerae expresses iron-regulated outer membrane proteins in vivo; Sciortino CV et al.; A comparison was made, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, of the outer membrane proteins of four strains of Vibrio cholerae grown in vivo in infant rabbits and in vitro in low-iron and iron-supplemented defined media . In vivo-grown V . cholerae expressed novel outer membrane-associated proteins which, in part, were similar to those observed on V . cholerae grown in vitro under conditions of iron deprivation. Infect Immun, 1983 Dec, 42(3), 1187 - 90 Differential complement activation and susceptibility to human serum bactericidal action by Vibrio species; Tamplin ML et al.; The ability of Vibrio vulnificus to resist human serum bactericidal action and to activate human complement was compared with similar cultures of Vibrio cholerae and Vibrio parahaemolyticus . Both V . vulnificus and V . parahaemolyticus had similar survival rates in sera and were much more resistant to killing than was V . cholerae . In contrast, V . vulnificus activated significantly less serum complement than did V . cholerae and V . parahaemolyticus . The relative ability of V . vulnificus to survive in serum and activate less complement than other Vibrio spp . tested may be related to its ability to cause chronic tissue infections and septicemias. Aust J Exp Biol Med Sci, 1983 Dec, 61 ( Pt 6), 713 - 25 Enzyme-linked immunoassays for antibodies against Vibrio cholerae; Cooper GN et al.; Glutaraldehyde-treated V . cholerae organisms bind firmly to the surfaces of plastic microELISA plates, thus providing a stable immobilized antigen for use in enzyme-linked immunosorbent assays (ELISA) . Serum absorption and ELISA-inhibition experiments indicate that, in addition to detecting natural antibodies in normal rat serum, the immobilized antigen may be used to quantitate specific anti-V . cholerae antibodies induced in rats by injection of live organisms . Apart from serotypically specific anti-lipopolysaccharide (LPS) antibodies, the reaction with immobilized organisms seems to involve antibodies common to Inaba and Ogawa serotypes; it is suggested that these antibodies are primarily directed against antigens of the outer membrane protein complex of V . cholerae cells . These findings have led to the development of an indirect ELISA method which quantitates levels of antibodies that react with heat-sensitive surface antigens of V . cholerae without involvement of anti-LPS antibodies . When used in conjunction with the indirect LPS-ELISA, the test has been found to provide a more detailed description of the serum antibody responses of rats to parenteral injections of live V . cholerae than has been reported previously. J Neurocytol, 1983 Dec, 12(6), 921 - 38 Ganglioside localization on myelinated nerve fibres by cholera toxin binding; Ganser AL et al.; GM1 ganglioside has been localized on the surfaces of myelinated, peripheral nerve fibres by using immunofluorescence to detect cholera toxin receptors . Unfixed, mouse sciatic nerves were teased into individual, intact fibres in order to expose their extracellular surfaces . Cholera toxin binding sites were abundant at all nodes of Ranvier; they were scarce on the internodal fibre surfaces . The nodal receptors were resistant to various degradative enzymes, including trypsin and proteinase K . Proteases did not unmask receptors on the internodal surfaces . Exogenous GM1 successfully competed for the toxin binding sites on the fibres . From this evidence and the specificity of cholera toxin binding, we conclude that GM1 ganglioside is abundantly present on the membrane surfaces of peripheral nodes of Ranvier and is not present on the internodal Schwann cell surfaces in an appreciable amount . The patterns of fluorescence within the node suggest that the axon and Schwann cell structures are sites where GM1 is localized . Treatment of the teased fibres with Vibrio cholerae neuraminidase, which is known to reduce polysialogangliosides to the monosialoganglioside GM1, induced cholera toxin binding on the internodal Schwann cell surfaces . The induced receptors, as well as their precursors, were resistant to trypsin and proteinase K . We conclude that the internodal Schwann cell surface is rich in an unidentified polysialoganglioside(s) that can be converted to GM1 by neuraminidase. Eur J Immunol, 1983 Dec, 13(12), 990 - 4 Immune elimination of aging platelets by autologous monocytes: role of membrane-specific autoantibody; Khansari N et al.; Membrane-bound IgG was found only on old populations of platelets from normal individuals . This IgG could be dissociated from senescent cells by repeatedly heating the cells . Heat-eluted IgG (He-IgG) prepared from senescent red blood cells was capable of binding to either heat-treated old platelets or Vibrio cholerae neuraminidase (VCN)-treated young platelets, suggesting expression of a common age-dependent antigen on the senescent red blood cells and old platelets . We analyzed the role of membrane-bound IgG in the immune elimination of aging platelets by direct phagocytosis of different platelet subpopulations by autologous monocytes in vitro . While removal of He-IgG from old platelets inhibited their phagocytosis, preincubation of either heat-treated old or VCN-treated young platelets promoted phagocytosis of these cells by autologous monocytes . The phagocytosis of senescent cells required intact IgG on these cells . Either removal of Fc fragments from He-IgG or treatment of autologous monocytes with Fc fragments prior to the phagocytosis assay resulted in a marked reduction of phagocytosis (greater than 75%) . We conclude that Fc receptors on the monocytes and the presence of membrane-specific IgG are crucial elements for immune elimination of senescent platelets. Infect Immun, 1983 Dec, 42(3), 924 - 9 Mapping of chromosomal genes that determine the El Tor biotype in Vibrio cholerae; Green BA et al.; The El Tor biotype of Vibrio cholerae has several characteristics that differentiate it from the classical biotype of V . cholerae . Among these are production of soluble hemolysin(s), a cell-associated hemagglutinin for chicken erythrocytes, resistance to polymyxin B, and resistance to bacteriophages of Mukerjee group IV . In the present study, we located the determinants for hemolysin (hly), chicken erythrocyte hemagglutinin (cha), and polymyxin B resistance (pmx) on the genetic map of V . cholerae . Transposon-facilitated recombination was used to perform conjugal matings between El Tor donor strains and classical recipient strains of V . cholerae . Recombinants were selected that had inherited specific nutritional markers from the donor strains and streptomycin resistance from the recipient strains . The recombinants were tested for the presence or absence of the unselected donor markers hly, cha, and pmx . These three El Tor biotype markers were found to be closely linked to each other and were located between the pyrA-201 and his-2 loci on the genetic map of V . cholerae. Jpn J Med Sci Biol, 1983 Dec, 36(6), 315 - 23 Serological studies on Vibrio fluvialis; Shimada T et al.; The serology of 138 strains of Vibrio fluvialis was studied . Eighteen O-antigenic groups were defined among them and it was shown that the H antigens of all the strains were identical regardless of the biovar . The presence of mucoid antigen, which inhibits O agglutination, was found in some strains . As all O antisera for V . fluvialis contained some R antibody, all diagnostic O sera must be absorbed with R organisms before use . Some O antigens of V . fluvialis were identical with those of certain Vibrio cholerae serovars. Biochim Biophys Acta, 1983 Nov 30, 725(2), 325 - 31 Low-potential cytochrome b as an essential electron-transport component of menaquinone reduction by formate in Vibrio succinogenes; Unden G et al.; Incorporation of the electron-transport enzymes of Vibrio succinogenes into liposomes was used to investigate the question of whether, in this organism, a cytochrome b is involved in electron transport from formate to fumarate on the formate side of menaquinone . (1) Formate dehydrogenase lacking cytochrome b was prepared by splitting the cytochrome from the formate dehydrogenase complex . The enzyme consisted of two different subunits (Mr 110 000 and 20 000), catalyzed the reduction of 2,3-dimethyl-1,4-naphthoquinone by formate, and could be incorporated into liposomes . (2) The modified enzyme did not restore electron transport from formate to fumarate when incorporated into liposomes together with vitamin K-1 (instead of menaquinone) and fumarate reductase complex . In contrast, restoration was observed in liposomes that contained formate dehydrogenase with cytochrome b (Em = -224 mV), in addition to the subunits mentioned above (formate dehydrogenase complex) . (3) In the liposomes containing formate dehydrogenase complex and fumarate reductase complex, the response of the cytochrome b of the formate dehydrogenase complex was consistent with its interaction on the formate side of menaquinone in a linear sequence of the components . The low-potential cytochrome b associated with fumarate reductase complex was not reducible by formate under any condition . It is concluded that the low-potential cytochrome b of the formate dehydrogenase complex is an essential component in the electron transport from formate to menaquinone . The low-potential cytochrome b of the fumarate reductase complex could not replace the former cytochrome in restoring electron-transport activity. J Biol Chem, 1983 Nov 25, 258(22), 13722 - 6 Nucleotide sequence analysis of the A2 and B subunits of Vibrio cholerae enterotoxin; Lockman H et al.; We have determined the sequence of the DNA encoding the A2 (gamma) and B subunits of Vibrio cholerae enterotoxin . The order of the subunits as they would be transcribed is A2-B and the termination codon of the A2 subunit overlaps the initiation codon of the B subunit by four bases . Sequence analysis revealed a region capable of coding for a 21-amino acid leader peptide located at the NH2 terminus of the B subunit . While the nucleotide sequence homology between the cholera enterotoxin subunits and the analogous sequences of the genes for Escherichia coli heat-labile enterotoxin (LT) was 72 and 77% for the A2 and B subunits, respectively, the predicted amino acid sequences of the A2 subunits were less similar . Twenty-nine of 46 (63%) amino acids of the A2 subunits and 98 of 124 (79%) amino acids of the B subunits were identical between cholera enterotoxin and LT . The predicted amino acid sequence of the enterotoxin from a V . cholerae El Tor biotype strain reported here differs from the previously published amino acid sequences of the toxin from a classical biotype of V . cholerae at seven residues . Comparison of the A2 and B amino acid sequences among El Tor and classical biotypes of V . cholerae and E . coli LT demonstrates two regions of highly conserved sequences: 12 and 22 uninterrupted amino acids are the same among the A2 and B subunits, respectively, from the three strains. Carbohydr Res, 1983 Nov 11, 123(1), 117 - 22 Hydrolytic release, and identification by g.l.c.-m.s., of 3-deoxy-D-manno-2-octulosonic acid in the lipopolysaccharides isolated from bacteria of the Vibrionaceae; Banoub JH et al.; The identification of the peracetylated methyl glycosides of 3-deoxy-D-manno-2-octulosonic acid (KDO) methyl esters was achieved by g.l.c.-m.s . These peracetylated methyl glycoside methyl esters were obtained from fully acetylated lipopolysaccharides and core oligosaccharides of representative strains of the Vibrionaceae family by the following sequence of mild reactions: acetolysis, methanolysis, and acetylation . KDO was shown to be present in all of the lipopolysaccharides (LPS), a result in direct contrast to the generally accepted view of the absence of this compound in LPS from this family of bacteria. Bull Soc Pathol Exot Filiales, 1983 Nov, 76(5), 644 - 51 {Diagnosis of Vibrio cholerae in the laboratory}; Dodin A et al.; The authors describe a new specific and immunologic process for V . cholerae isolation . Specific anti-V . cholerae antibodies (specific IgG anti-fraction Ch 1 + 2) are sticked on magnetic beads . These beads are added to choleric stool . V . cholerae germs stick on the beads . These particles are drawn out of the mixture with the help of a magnet and laid on a Mueller-Hinton agar plate . Incubation is carried out for 18 hours . One drop of toluen is then added on the beads . Wells are sinked in the agar plate and filled with specific serum or antitoxin serum . 12 hours after, the appearance of a precipitation line, facing the specific serum, reveals the presence of toxinogen V . cholerae. Appl Environ Microbiol, 1983 Nov, 46(5), 1232 - 3 Vibrio cholerae (non-O1) isolated from California coastal waters; Kenyon JE et al.; Nineteen strains of Vibrio cholerae non-O1 were isolated from five separate marine sites along the Santa Cruz County coast . This environmental study was initiated after a human case of non-O1 cholera-like diarrhea was acquired endemically. Infect Immun, 1983 Nov, 42(2), 501 - 9 Production of cholera toxin-like toxin by Vibrio mimicus and non-O1 Vibrio cholerae: batch culture conditions for optimum yields and isolation of hypertoxigenic lincomycin-resistant mutants; Spira WM et al.; Vibrio mimicus 61892, isolated in 1977 from a case of watery diarrhea in Bangladesh, produces an enterotoxin which possesses activity in Y-1 mouse adrenal cells and in rabbit ileal loops which is identical to the prototype cholera toxin (CT) produced by Vibrio cholerae 569B . The neutralization of the adrenal cell activity of 61892 toxin and 569B CT by homologous and heterologous antisera generates parallel titration curves which show complete neutralization in all cases . Paired titrations in the ganglioside GM1 enzyme-linked immunosorbent assay (using either CT or Escherichia coli heat-labile toxin antitoxin) of both toxins indicates that 61892 toxin is antigenically indistinguishable from 569B CT . The specific activity of the two toxins in the rabbit ileal loop is virtually identical . Batch culture production of CT-like toxin and CT by isolates of V . mimicus and different biotypes of V . cholerae was found to be highest in shake flask cultures of Casamino Acids-yeast extract broth grown at 27 degrees C with vigorous aeration . Incorporation of lincomycin into the growth medium at a concentration of 50 micrograms/ml increased yields from wild-type strains . Dramatically higher yields were obtained when a spontaneous resistance mutant of strain 61892 was grown in the presence of 200 to 300 micrograms of lincomycin per ml . Under these conditions, yields of CT-like toxin were increased by 300- to 500-fold, and the highest yields reached more than 100 micrograms/ml after 44 h of culture . This is substantially higher than that reported in the literature for CT production by any strain of V . cholerae, including hypertoxigenic strain 569B. Cell, 1983 Nov, 35(1), 253 - 63 Duplication and amplification of toxin genes in Vibrio cholerae; Mekalanos JJ; Vibrio cholerae strains of the classical biotype all contain two widely separated copies of the cholera toxin operon ctxAB . In contrast, EI Tor strains containing multiple copies of ctx have their copies arranged on large tandem repeats which are either 7 or 9.7 kb in length . The variation in size among these large tandem duplications was due to a difference in the copy number of a smaller, 2.7 kb, tandemly repeated sequence (RS1) that is located at the novel joint of these duplications, as well as upstream and downstream of ctx . Southern blot hybridization analysis indicated that amplification of a DNA region carrying ctx and flanked by direct repeats of RS1 may be responsible for the hypertoxinogenic phenotype of EI Tor variants selected by intraintestinal growth in rabbits. Infect Immun, 1983 Nov, 42(2), 639 - 44 Vibrio cholerae soluble hemagglutinin/protease is a metalloenzyme; Booth BA et al.; A soluble hemagglutinin/protease produced by Vibrio cholerae, which has previously been shown to hydrolyze fibronectin and ovomucin and to cleave lactoferrin and the A subunit of the heat-labile enterotoxin of Escherichia coli, appears to be a zinc metalloendopeptidase . Both its hemagglutinative and protease functions are inhibited by chelating agents, including Zincov, a hydroxamic acid derivative specifically designed to inhibit zinc metalloproteases . Thermolysin, a known zinc-containing protease, also causes hemagglutination of responder chicken erythrocytes . This activity is inhibited by Zincov, which does not affect the hemagglutination activity of trypsin and pronase . The hemagglutinin/protease is active on furylacryloyl-Gly-Leu-NH2, a synthetic substrate for thermolysin and other similar proteases . The hemagglutination activity of V . cholerae-infected or cholera toxin-treated infant rabbit intestinal fluid is not inhibited by Zincov, which suggests that this activity is not due to the hemagglutinin/protease, as formerly proposed. Can J Microbiol, 1983 Nov, 29(11), 1481 - 6 Mechanism of poliovirus inactivation by cell-free filtrates of marine bacteria; Toranzo AE et al.; The mechanism of enterovirus inactivation by marine bacteria was investigated using poliovirus type 1 as a model virus and with strains of Pseudomonas and Vibrio isolated from the marine environment . Treatment of virus with cell-free filtrates from late log phase bacterial cultures produced alterations in the viral capsid as shown by a reduction in efficiency of adsorption to host cells, increased sensitivity to ribonuclease, and by the release of ribonucleic acid from the treated virions . Filtration of 14C-labelled, treated virus through 25-nm filters revealed that the majority of the isotope (85-96%) passed the filters, indicating extensive capsid disruption . However, the most rapid and pronounced change observed during virus inactivation was the loss of infectivity, suggesting that enzymatic degradation is not the first event in the poliovirus inactivation process by marine bacteria. J Bacteriol, 1983 Nov, 156(2), 880 - 7 Iron uptake system medicated by Vibrio anguillarum plasmid pJM1; Walter MA et al.; Plasmid pJM1 from an invasive strain of Vibrio anguillarum mediates an iron-sequestering system that is associated with the ability of this bacterium to cause septicemia in marine fishes . This plasmid-mediated iron uptake system was analyzed by using mutations caused by transposon Tnl . Restriction endonuclease analysis of iron uptake-deficient and -proficient derivatives generated by insertion of Tnl and molecular cloning experiments permitted us to localize the plasmid regions involved in the process of iron sequestration to a stretch of about 20 kilobase pairs . In addition, the existence of two plasmid-mediated components involved in the process of iron uptake in V . anguillarum was defined: a diffusible substance which functions as a siderophore and a nondiffusible receptor for complexes of iron-siderophore, which we have tentatively identified as the pJM1 plasmid-mediated outer membrane protein OM2 of V . anguillarum. J Infect Dis, 1983 Nov, 148(5), 931 - 9 Prophylactic significance of the nonlipopolysaccharide antigens of Vibrio cholerae; Attridge SR et al.; The infant-mouse cholera model was used for evaluation of the immunoprophylactic significance of the nonlipopolysaccharide antigens of Vibrio cholerae . The protective efficiency of antibodies to the nonlipopolysaccharide components of the 569B strain was much greater than that of antibodies to the lipopolysaccharide . Protective nonlipopolysaccharide antigens were not detected in two other strains, however; the possible basis for this restricted distribution was considered in light of studies from other laboratories . An investigation of the mechanism by which antibodies mediate protection in this model suggested that the critical property of protective antibodies is their capacity to block vibrio attachment directly. Biochim Biophys Acta, 1983 Oct 31, 725(1), 41 - 8 Structural properties of the proteoliposomes catalyzing electron transport from formate to fumarate; Unden G et al.; The electron-transport chain catalyzing fumarate reduction by formate has recently been reconstituted from the formate dehydrogenase complex and the fumarate reductase complex from Vibrio succinogenes, in a liposomal preparation containing vitamin K-1 (Unden, G . and Kroger, A . (1982) Biochim . Biophys . Acta 682, 258-263) . We have now investigated the structural properties of this preparation . The preparation was found to consist of a homogeneous population of unilamellar proteoliposomes with an average diameter of about 100 nm and an internal volume of 2-4 ml/g phospholipid . The buoyant density (1.07 g/ml) was consistent with the protein/phospholipid ratio (0.2 g/g) of the preparation . Leakage of glucose from the internal spaces of the proteoliposomes was negligibly slow . Proteoliposomes prepared with either of the enzyme complexes showed peripheral projections mainly on the outer surface, when examined by electron microscopy after negative staining . The size, orientation and surface density of the projections were consistent with those of the enzymes . Most of the substrate and dye-reactive sites (70-90%) of the enzymes in the proteoliposomes were accessible to external non-permeant substrates . The proteoliposomes catalyzing electron transport were formed by freeze-thawing a mixture of liposomes and protein-phospholipid complexes which did not perform electron transport from formate to fumarate . Nearly the entire amount of the enzymes supplied (0.2 g protein/g phospholipid) was incorporated into the liposomes by this procedure . The transformation of liposomes into proteoliposomes was accompanied by exchange of the internal solutes with the external medium. Carbohydr Res, 1983 Oct 28, 122(2), 209 - 16 Studies on the partial structure of the O-antigen of Vibrio cholera Ogawa G-2102; Majumder M et al.; Detailed information was obtained regarding the partial structure of the lipopolysaccharide (LPS), containing glucose, glucuronic acid, 2-amino-2-deoxy-glucose, L-glycero-D-gluco-heptose, and small proportions of L-glycero-D-manno-heptose, mannose, and galactose, isolated from Vibrio cholera Ogawa G-2102 . Structures of three oligosaccharides were determined . Results of deamination experiments established the sequence of the linkages between the amino sugar and heptose residues in the O-antigenic polysaccharide. Biochim Biophys Acta, 1983 Oct 12, 734(2), 368 - 72 The effect of neuraminidase on the relative surface charge-associated properties of rat red blood cells of different ages; Walter H et al.; Approx . 70% of the sialic acid on the rat erythrocyte surface is susceptible to cleavage by neuraminidase (Vibrio cholerae) . Neuraminidase treatment results in a reduction in the partition coefficient (K) of the red cells in a charged dextran-poly(ethylene glycol) aqueous phase system and in the electrophoretic mobility of the cells . Countercurrent distribution of rat neuraminidase-treated erythrocytes, containing 59Fe-labeled mature red cells of distinct age, indicates that (a) the electrophoretic mobilities of red cells in different cavities along the extraction train increase with increasing K, as is the case with untreated erythrocytes, and (b) the cell age-related differences in surface charge-associated properties are neither eliminated nor altered by the enzyme action. FEBS Lett, 1983 Oct 3, 162(1), 177 - 9 Synthesis of phage-specific transfer RNA molecules by vibriophage phi 149; Ghosh RK et al.; 32P-Labelled tRNA was isolated from uninfected and phage phi 149-infected Vibrio cholerae cells . These tRNA preparations were then hybridised with DNA isolated from phage phi 149 . Significant hybridisation was observed only with tRNA from phage phi 149-infected cells . This strongly suggests that infection of classical vibrio with phage phi 149 results in the synthesis of phage-specific tRNA molecules. J Gen Microbiol, 1983 Oct, 129 (Pt 10), 3185 - 96 Isolation and characterization of the outer membrane from Vibrio parahaemolyticus; Koga T et al.; The outer membrane of Vibrio parahaemolyticus strain 3283-61 (serotype O2:K3) was isolated from blebs released upon spheroplast formation, in the presence of lysozyme and EDTA, by isopycnic sucrose density gradient centrifugation . SDS-PAGE of the outer membrane fraction prepared from cells grown in nutrient broth containing 3% (w/v) NaCl revealed five major proteins, designated a to e, with apparent approximate molecular weights: a, 44 000; b, 36 000; c, 33 500; d, 26 500; e, 22 000 . An increase in NaCl concentration in the growth medium resulted in an increase of proteins b and c, whereas a decrease to 0.5% (w/v) induced two additional major proteins with respective molecular weights of about 35 000 and 32 000 . Proteins a and b appeared to be loosely associated with the peptidoglycan layer since they were largely retained after extraction with 2% (w/v) SDS at 50 degrees C for 30 min . Proteins c and/or e may play a role in phage VP1-receptors since phage-resistant mutants derived from strain 3283-61 had significantly diminished amounts of both proteins . The major outer membrane proteins varied in number and molecular weight in strains of V . parahaemolyticus belonging to different K-serotypes. J Appl Bacteriol, 1983 Oct, 55(2), 263 - 82 A numerical taxonomic study of species of Vibrio isolated from the aquatic environment and birds in Kent, England; West PA et al.; A numerical taxonomic study has been carried out to confirm the identity of strains of the family Vibrionaceae isolated during an ecological study . A total of 237 strains were studied including 148 from the aquatic environment, 6 from estuarine birds, 1 from sheep faeces, and 61 control cultures . Duplicates of 21 of the strains were randomly selected and included to estimate test and operator error . Taxonomic resemblance was estimated on the basis of 148 characters using Euclidean distance . The taxonomic position of some strains was reevaluated using the pattern different coefficient . Strains were clustered by three methods, all of which gave similar results . The estimated average probability of test error was 1.5% . Strains previously identified as Vibrio anguillarum fell into four distinct phenons corresponding to V . anguillarum biovar I, 'V . anguillarum biovar II', V . diazotrophicus, and strains pathogenic to oyster larvae . The latter group characteristically degraded xanthine and probably represents a new species . The phenon corresponding to V . cholerae included the type strain, strains of human origin, and strains isolated in the United Kingdom from birds and the aquatic environment . Some strains of V . cholerae were luminous . Other phenons were identified as V . metschnikovii, V . fluvialis, and Aeromonas spp. Hoppe Seylers Z Physiol Chem, 1983 Oct, 364(10), 1419 - 29 Binding and phagocytosis of sialidase-treated rat erythrocytes by a mechanism independent of opsonins; Muller E et al.; Rat peritoneal macrophages bind and phagocytoze homologous sialidase-treated erythrocytes at a rate which is dependent on the amount of sialic acid that has been removed from the cells . Increased binding of erythrocytes is observed after the removal of 10-20% of membrane sialic acid, while for phagocytosis at least 30-40% of this substance must be removed . With Vibrio cholerae sialidase only a partial (80%) hydrolysis of rat erythrocyte sialic acid is possible, whereas Arthrobacter ureafaciens sialidase leads to complete desialylation and therefore causes stronger binding and phagocytosis of the erythrocytes than the V . cholerae enzyme . Preincubation of peritoneal macrophages with sialidase impairs binding and phagocytosis . Experiments were performed to account for the stimulation of binding and phagocytosis observed in the presence of native, homologous serum . However, an involvement of immunoglobulins and complement factors of the classical and alternative pathway in the engulfment process has been excluded . Fibronectin, tuftsin and substance P have no influence, either . On the other hand, peanut agglutinin and Erythrina crystagalli agglutinin are potent stimulators of binding and phagocytosis of sialidase-treated erythrocytes, whereas soybean agglutinin has only little and limulin no influence at all . It is concluded that sialidase-treated erythrocytes, having been bound to the beta-galactose-specific lectin on the macrophage surface, are phagocytozed as a function of their number and binding strength to the macrophages . The influence of native serum and especially of the plant lectins on this process is discussed. J Clin Microbiol, 1983 Oct, 18(4), 816 - 24 Vibrio furnissii (formerly aerogenic biogroup of Vibrio fluvialis), a new species isolated from human feces and the environment; Brenner DJ et al.; Strains formerly classified as the aerogenic (gas-producing) biogroup of Vibrio fluvialis were shown by DNA relatedness to be a separate species . The species was named Vibrio furnissii sp . nov . (type strain ATCC 35016 = CDC B3215) . Three strains of V . furnissii were 79% or more related to the type strain of V . furnissii and about 50% related to the type strain of V . fluvialis . V . fluvialis strains were 40 to 64% related to the type strain of V . furnissii . Divergence in related sequences was only 0.0 to 1.5% among strains of V . furnissii and among strains of V . fluvialis but was 5.0 to 8.0% in interspecific reactions between V . fluvialis and V . furnissii . V . furnissii was aerogenic (produced gas from the fermentation of carbohydrates), whereas V . fluvialis was anaerogenic (did not produce gas from the fermentation of carbohydrates) . Another test of some help in differentiating the two species was fermentation of L-rhamnose (57% positive for V . furnissii and negative for V . fluvialis) . In addition to the reactions above, V . furnissii is distinguished from other salt-requiring vibrios on the basis of its positive reactions in tests for Moller L-arginine, L-arabinose, maltose, and D-mannitol and its negative reactions for Moller L-lysine and L-ornithine, lactose, and Voges-Proskauer . V . furnissii has been isolated from patients with acute gastroenteritis in at least two outbreaks of food poisoning; its role as a cause of diarrhea needs further study. Ann Intern Med, 1983 Oct, 99(4), 558 - 9 Vibrios on the half shell: what the walrus and the carpenter didn't know; Blake PA; At least nine Vibrio species have been associated with disease in the United States . Vibrio fluvialis, V . hollisae, V . mimicus, and V . parahaemolyticus cause diarrheal diseases, but may also be encountered in extraintestinal infections such as wound and ear infections, septicemia, and cholecystitis . Vibrio alginolyticus, V . damsela, V . metschnikovii, and V . vulnificus primarily cause extraintestinal disease . Toxigenic V . cholerae O1 is the cause of epidemic cholera, whereas nontoxigenic V . cholerae O1 and non-O1 V . cholerae have been associated with both diarrheal and extraintestinal diseases . Most reports of vibrio infections have come from states along the Atlantic Ocean and Gulf of Mexico and from Hawaii, and most of the infections have occurred during summer and fall . Wound and ear infections have occurred after exposure to salty or brackish water or to drippings from raw seafoods . Foodborne vibrio infections are almost all caused by seafoods, especially oysters eaten raw . Thorough cooking and careful handling will render seafoods safe for consumption. Ann Intern Med, 1983 Oct, 99(4), 464 - 9 Spectrum of Vibrio infections in a Gulf Coast community; Bonner JR et al.; Infection with Vibrio species was diagnosed in 23 patients over a 10-year period in a Gulf Coast community . Species isolated were Vibrio vulnificus, V . cholerae, V . parahaemolyticus, and V . alginolyticus . Clinical presentations included diarrhea, wound infection, and bacteremia . Gastrointestinal tract infection was infrequently diagnosed, despite the routine use of stool culture media appropriate for vibrio isolation . Bacteremic infections were severe and occurred only in patients with underlying diseases . Wound infections were complicated by tissue necrosis and required surgical debridement . In patients with underlying diseases, soft tissue infections often progressed to fatal septicemias. Infect Immun, 1983 Oct, 42(1), 224 - 30 Molecular epidemiological studies of United States Gulf Coast Vibrio cholerae strains: integration site of mutator vibriophage VcA-3; Goldberg S et al.; Environmental and clinical Vibrio cholerae O-1 strains isolated from the U.S . Gulf Coast region were found to be lysogenic for a vibriophage which we have designated VcA-3 . Comparison of VcA-3 with the previously described vibriophages VcA-1 and VcA-2 has shown that VcA-1 and VcA-3 are homoimmune, have extensive sequence homology, but have markedly different restriction endonuclease digestion patterns . VcA-3 was found to randomly integrate into the V . cholerae RV79 chromosome and to introduce stable auxotrophic mutations . We show that all U.S . Gulf Coast environmental and clinical isolates that are lysogenic for VcA-3, including both tox+ and tox- isolates, contain the prophage integrated at an identical chromosomal site . Given the known stability of temperate mutator bacteriophages, these results suggest that there is a clonal relationship among the V . cholerae O-1 strains examined in this study, including the tox+ and tox- isolates. Histochem J, 1983 Oct, 15(10), 987 - 97 Sialomucin in Paget cells of extramammary Paget's disease; Sasai Y et al.; The cytoplasmic sialomucin in Paget cells of extramammary Paget's disease was examined by means of a battery of histochemical techniques . The staining methods used involved an electrolyte--Alcian Blue (pH 5.8), periodic acid--Schiff and azure A at selected pH levels . Methylation, saponification, borohydride reduction, acid hydrolysis, and digestion with diastase, neuraminidase (Vibrio cholerae) or chondroitinase ABC, were also employed . The cytoplasmic mucin was found to exhibit positive reaction for the above staining which were variously altered by the chemical modification procedures and diminished in intensity or abolished by digestion with neuraminidase . These results suggest that the cytoplasmic mucin is sialomucin without side-chain substituent in extramammary Paget's disease located in axillary or genital area, and with a substituent at C7 in the disease located in perianal area. Infect Immun, 1983 Oct, 42(1), 427 - 30 Evidence indicating that the cholera toxin structural genes of Vibrio cholerae RJ1 and 3083-2 are between met and trp; Saunders DW et al.; An enterotoxin A subunit-specific radioactive probe was used to correlate expression of vct-1 or vct-2 with the presence of specific restriction fragments in the DNAs of several wild-type and recombinant Vibrio cholerae strains . The data are consistent with the conclusion that vct is a cholera toxin structural gene. Hoppe Seylers Z Physiol Chem, 1983 Oct, 364(10), 1411 - 7 On the specificity of sialidase . Synthesis and properties of N5-acetyl-beta-D-neuraminoylpeptides - AcNeu-Gly-OH, AcNeu-Glu-OH, AcNeu-Phe-OH - and the corresponding alpha-ketosides; Eschenfelder V et al.; By means of the mixed anhydride procedure the benzyl alpha-ketoside of N5-acetyl-D-neuraminic acid was linked to L-glycine, L-glutamic acid and L-phenylalanine . Hydrogenolytic cleavage of the benzyl group resulted in the corresponding free N5-acetyl-beta-D-neuraminoylpeptides . This new class of compounds is no substrate for Vibrio cholerae sialidase . The enzyme does not split the benzyl alpha-ketosides of N5-acetyl-D-neuraminoylpeptides nor is its activity inhibited by these compounds . The results strongly support the assumption that in sialidase substrates the carboxy group must be located close to the ketosidic oxygen . N-(N5-acetyl-beta-D-neuraminoyl)-L-phenylalanine was readily hydrolysed by carboxypeptidase A from bovine pancreas. J Immunol Methods, 1983 Sep 30, 63(1), 25 - 34 A rapid and sensitive assay for neuraminidase using peanut lectin hemagglutination: application to Vibrio cholera and Trypanosoma cruzi; Pereira ME; A neuraminidase assay based on peanut lectin agglutination is described . Human red blood cells are used both as substrate for the enzyme and as probe for the lectin . The validity of the method is ascertained by measuring the enzyme and lectin activities on erythrocytes whose outer membrane sialic acid was labeled with tritium after oxidation with sodium periodate followed by reduction with sodium borotritiide . The neuraminidases of Trypanosoma cruzi and Vibrio cholera are used as examples; in both cases, a linear relationship is observed between the degree of erythrocyte desialylation and the peanut hemagglutination titer . For the hemagglutination assay, lectin in homogeneous form as well as in crude peanut extracts may be used, and free sialic acid need not be separated from substrate-bound sialic acid . The hemagglutinating activity of peanut lectin is not affected by pre-treatment of the erythrocytes with various proteases . The method is particularly useful in the neuraminidase analysis of multiple samples, such as in gel filtration chromatography and for screening of hybridoma antibodies against neuraminidase. S Afr Med J, 1983 Sep 10, 64(11), 405 - 6 Vibrio cholerae bacteraemia in a newborn infant . A case report; Coovadia YM et al.; A 6-day-old Black male infant presented with diarrhoea and biochemical evidence of severe electrolyte imbalance . Despite treatment with intravenous fluids and antibiotics, he died within 24 hours of admission . Enterotoxigenic Vibrio cholerae, biotype E1 Tor, serotype Inaba, was isolated from the blood . The significance of this finding is discussed. N Engl J Med, 1983 Sep 1, 309(9), 523 - 6 Cholera on a Gulf Coast oil rig; Johnston JM et al.; A single case of severe diarrhea on a floating Texas oil rig was followed two days later by what proved to be the largest outbreak of cholera in the United States in over a century . After isolation of toxigenic Vibrio cholerae El Tor Inaba of the typical United States phage type from the index patient's stool, the ensuing investigation detected 14 additional cases of cholera and one asymptomatic infection serologically . Infection was associated with eating rice on the oil rig on a particular day (P = 0.03) when an open valve permitted the rig's drinking-water system to be contaminated by canal water containing sewage (including that from the index patient) discharged from the rig . The rice had been rinsed in the contaminated water after cooking, and before being served it had been maintained at a temperature that allows V . cholerae 01 to multiply . Toxigenic V . cholerae 01 is persisting in the United States, and large common-source outbreaks of cholera can occur if proper sanitation is not maintained. Diagn Microbiol Infect Dis, 1983 Sep, 1(3), 229 - 32 Bacterial flora of marine penetrating injuries; Pien FD et al.; Bacteriologic and clinical features of 28 cases of traumatic marine injuries are described . The most common bacterial isolates were normal skin bacteria, Staphylococcus aureus and, in 11% of cases, Vibrio alginolyticus . All injuries responded to local wound care, and only half received antibiotic therapy. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1983 Sep, 255(2-3), 285 - 93 Studies on bacteriocin production by NAG-strains of Vibrio cholerae as a possible epidemiological marker; Israil AM et al.; 201 "non-agglutinable" - i.e . serogroup O:1-negative - Vibrio cholerae strains were tested for bacteriocin production using 14 of the Abbott-shannon standard set of colicin-sensitive strains as indicators . 41.2% of the strains proved to be bacteriocin producers . 29 out of 67 (43%) vibriocin-positive strains originating from one large geographical region gave the same reaction pattern with the applied indicator strains . The possible significance of these vibriocins for epidemiological studies is discussed, and the necessity for further investigations prior to routine bacteriocin typing of O:1-negative V . cholerae strains is pointed out. Zh Mikrobiol Epidemiol Immunobiol, 1983 Sep, (9), 89 - 92 {Phage typing of nonagglutinating vibrios isolated in the territory of the USSR}; Ganin VS et al.; The work presents the results of the phage typing of 2,437 NAG vibrio strains isolated in the USSR from patients and carriers (374 strains), from open water basins (1,675 strains), from sewage (234 strains) and from bottom silt (172 strains) . Phage typing was carried out with a set of 5 phages capable of lyzing enteropathogenic NAG vibrios (phages TEPV 1, 2, 3, 4, 5) . NAG vibrios isolated from humans were sensitive to phages TEPV in 45% of cases and NAG vibrios isolated from the environment, only in 29.9% of cases . Enteropathogenic NAG vibrios belonging to phagovar 1 were isolated most frequently from samples taken both from humans and from the environment . The NAG vibrios isolated from patients, carriers and the sources of water supply nearest to them belonged to the same phagovars. J Clin Microbiol, 1983 Sep, 18(3), 480 - 5 Monoclonal antibodies against group- and type-specific lipopolysaccharide antigens of Vibrio cholerae O:1; Gustafsson B et al.; Hybrid cell lines producing monoclonal antibodies against the O-antigenic determinants of Vibrio cholerae O:1 have been established . The specificity of the antibodies was ascertained by enzyme-linked immunosorbent assay inhibition experiments by using lipopolysaccharides from V . cholerae O:1 strains and type strains of groups O:2 and O:21 . The anti-A antibody was of the immunoglobulin M (IgM) class, whereas the anti-B and -C antibodies were IgG3 . The antibodies had a good agglutinating capacity when tested against V . cholerae O:1 strains in the slide agglutination test. J Virol, 1983 Sep, 47(3), 385 - 91 Biological consequences of neuraminidase deficiency in Newcastle disease virus; Smith GW et al.; A second-step revertant (L1) of a temperature-sensitive mutant (C1) of Newcastle disease virus agglutinated erythrocytes normally but had less than 3% of the wild-type (strain AV) levels of neuraminidase activity . Revertant L1 had seven times more virion-associated N-acetylneuraminic acid (NANA) than strain AV . NANA residues on purified virions were specifically labeled with periodate and tritiated borohydride . Analyses of radiolabeled L1 virions on sodium dodecyl sulfate-polyacrylamide gels showed that most of the virion-associated NANA was in a high-molecular-weight component with an electrophoretic mobility different from that of any known viral protein . NANA was also detected in molecules with the electrophoretic mobility of the viral glycoproteins HN and F1 . Revertant L1 had a twofold lower rate constant of attachment to HeLa cells than that of the wild-type . Treatment of L1 virions with Vibrio cholerae neuraminidase removed the excess NANA and returned L1 attachment kinetics to normal . Revertant N1, which has 10-fold more neuraminidase activity than L1, penetrated host cells at the same rate as L1 . L1 was impaired in elution from erythrocytes . Removal of virion-associated NANA exacerbated this defect . Despite a small disadvantage in attachment and a major defect in elution relative to strain AV, revertant L1 enjoyed a slight advantage over the wild-type during a single reproductive cycle in cultured chicken embryo cells. Infection, 1983 Sep-Oct, 11(5), 272 - 4 A cholera-like illness in a traveller due to a mixed infection with enterotoxigenic Escherichia coli, Vibrio parahaemolyticus and Pseudomonas aeruginosa; Bockemuhl J et al.; A healthy 67-year-old male traveller developed a cholera-like disease after returning from a five-week stay in Pakistan and India . In addition to Vibrio parahaemolyticus and large numbers of Pseudomonas aeruginosa, two strains of enterotoxigenic Escherichia coli were isolated from a single stool specimen. Infect Immun, 1983 Sep, 41(3), 896 - 901 Evidence that a non-O1 Vibrio cholerae produces enterotoxin that is similar but not identical to cholera enterotoxin; Yamamoto K et al.; Cholera-like enterotoxin produced by a non-O1 strain of Vibrio cholerae, S7 (S7 enterotoxin), isolated from human diarrheal stool, was purified, and its physicochemical, biological, and immunological properties were compared with those of cholera enterotoxin from V . cholerae O1 569B (CT) and an enterotoxin produced by another non-O1 V . cholerae (E8498 enterotoxin) reported previously (Yamamoto et al., Infect . Immun . 39:1128-1135, 1983) . The purified S7 enterotoxin had physicochemical properties different from those of CT and E8498 enterotoxin . S7 enterotoxin had greater relative mobility in conventional polyacrylamide gel disc electrophoresis and a lower isoelectric point, and its B subunit was smaller than those of CT and E8498 enterotoxin . The results of sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis suggested that the size of the aggregate of the B subunits of S7 enterotoxin is larger than that of CT and E8498 enterotoxin . The biological and immunological properties of S7 enterotoxin were indistinguishable from those of CT and E8498 enterotoxin . These results indicate that non-O1 vibrios may produce more than one kind of cholera-like enterotoxin: one which is identical to CT (E8498 enterotoxin type) and another which is not identical to CT (S7 enterotoxin type). Appl Environ Microbiol, 1983 Sep, 46(3), 704 - 9 Enterotoxin production by Vibrio cholerae and Vibrio mimicus grown in continuous culture with microbial cell recycle; Spira WM et al.; We have examined the effect of complete cell recycle on the production of cholera toxin (CT) by Vibrio cholerae and CT-like toxin by Vibrio mimicus in continuous culture fermentations . Complete cell recycle was obtained by filtering culture fluids through Amicon hollow fibers with an exclusion limit of 100,000 daltons (H1P100-20) and returning the concentrated cell slurry to the fermentor . A single 1-liter laboratory fermentor system modified with this recycle loop was capable of producing over 20 liters of cell-free culture filtrate per day . Toxin production in this system was compared with yields obtained in traditional continuous cultures and in shake flask cultures . Yields of CT from V . cholerae 569B in the recycle fermentor were highest at the highest dilution rate employed (1.0 vol/vol per h) . The use of complete cell recycle dramatically increased yields over those obtained in continuous culture and equaled those obtained in shake flasks . The concentration of CT in the filtrate was slightly less than half of that measured in culture fluids sampled at the same time . Similarly, V . mimicus 61892 grown in the presence of 50 micrograms of lincomycin per ml produced 280 ng of CT per ml in the recycle fermentor, compared with 210 ng/ml in shake flasks under optimal conditions . The sterile filtrate from this fermentation contained 110 ng/ml. Zh Mikrobiol Epidemiol Immunobiol, 1983 Sep, (9), 24 - 8 {Chemotaxis of cholera vibrios: a study method and the relation to different substances}; Dimitrova NI et al.; The chemotaxis of V . cholerae in response to 56 different substances (amino acids, carbohydrates, salts, etc.) has been studied by the methods of visual observation and quantitative determination . Attractants, neutral substances and one repellent have been revealed . Adler's method (1973) has been modified with regard to the requirements for the working procedures in handling the causative agents of highly dangerous infections. FEBS Lett, 1983 Aug 22, 160(1-2), 31 - 6 Uptake of adenosine in a marine bacterium is not an active transport process; Bengis-Garber C; The uptake and intracellular interconversions of {8-14C}adenosine in a marine bacterium Vibrio harveyi were investigated under varying physiological conditions . The results indicated that in contrast with the current views, translocation of adenosine across the cytoplasmic membrane in Vibrio harveyi was not driven by respiration . The uptake of adenosine was dependent upon its intracellular utilization and was inhibited under conditions preventing its metabolic conversions. J Am Acad Dermatol, 1983 Aug, 9(2), 262 - 8 Culture of Vibrio extorquens from severe, chronic skin ulcers in a Puerto Rican woman; Lambert WC et al.; A 48-year-old Puerto Rican woman developed extensive ulcers on her buttocks, right arm, and thighs over a 3 1/2-year period . The lesions began as small, subcutaneous nodules which subsequently ulcerated and expanded up to 19 cm in diameter . Biopsy of both ulcerated and nonulcerated lesions showed acid-fast bacilli . Culture of both types of lesions grew Vibrio extorquens, a partially acid-fast methanolophilic organism not previously associated with disease in humans . The patient developed agglutinating antibody, titer 1:80, to this organism . The disease responded to treatment with antibiotics to which the organism was sensitive in vitro. J Am Acad Dermatol, 1983 Aug, 9(2), 243 - 51 Vibrio vulnificus infection . Case report and update since 1970; Wickboldt LG et al.; Vibrio vulnificus infections is being reported with increasing frequency in coastal regions of the United States . Raw seafood consumption, particularly raw oysters, and wounds acquired in a marine environment predispose to infection . Patients with advanced liver disease are at increased risk of developing septicemia . V . vulnificus is a virulent pathogen producing significant morbidity and mortality; its virulence relates in part to the production of exotoxin . Skin lesions occur early in the clinical course of infection and provide means of specific diagnosis . The patient and the consulting physician are well served by the dermatologist capable of recognizing this infectious disease. Ann Intern Med, 1983 Aug, 99(2), 169 - 71 Disease from infection with Vibrio mimicus, a newly recognized Vibrio species . Clinical characteristics and edipemiology; Shandera WX et al.; Clinical and epidemiologic characteristics of disease associated with a newly described nonhalophilic Vibrio species, Vibrio mimicus, were identified by studying isolates from 21 patients referred to the Centers for Disease Control between 1977 and 1981 . Two isolates were from the ears of patients with otitis who had recently been exposed to seawater . Nineteen isolates were from stool samples; these patients generally had diarrhea, nausea, vomiting, and abdominal cramps, with fever, headache, and bloody diarrhea occurring in fewer than half . Persons with diarrhea were more likely than age- and sex-matched controls to have eaten raw oysters (p = 0.013) . Although most cases were sporadic, three were associated with a single outbreak . Only two isolates produced toxin found by enzyme-linked immunosorbent assay or the Y-1 adrenal cell assay for heat-labile toxin, and none produced heat-stable toxin found by the infant mouse assay . Vibrio mimicus should be considered in the differential diagnosis of acute gastroenteritis occurring after recent ingestion of seafood (especially raw oysters) and in acute otitis after exposure to seawater. Ann Intern Med, 1983 Aug, 99(2), 169 - 71 Disease from infection with Vibrio mimicus, a newly recognized Vibrio species; Shandera WX et al.; Clinical and epidemiologic characteristics of disease associated with a newly described nonhalophilic Vibrio species, Vibrio mimicus, were identified by studying isolates from 21 patients referred to the Centers for Disease Control between 1977 and 1981 . Two isolates were from the ears of patients with otitis who had recently been exposed to seawater . Nineteen isolates were from stool samples; these patients generally had diarrhea, nausea, vomiting, and abdominal cramps, with fever, headache, and bloody diarrhea occurring in fewer than half . Persons with diarrhea were more likely than age- and sex-matched controls to have eaten raw oysters (p = 0.013) . Although most cases were sporadic, three were associated with a single outbreak . Only two isolates produced toxin found by enzyme-linked immunosorbent assay or the Y-1 adrenal cell assay for heat-labile toxin, and none produced heat-stable toxin found by the infant mouse assay . Vibrio mimicus should be considered in the differential diagnosis of acute gastroenteritis occurring after recent ingestion of seafood (especially raw oysters) and in acute otitis after exposure to seawater. J Gen Virol, 1983 Aug, 64 (Pt 8), 1749 - 55 Repair of ultraviolet light-induced DNA damage in cholera bacteriophages; Palit BN et al.; DNA repair-proficient and -deficient strains of Vibrio cholerae were used to examine host cell reactivation, Weigle reactivation and photoreactivation of u.v.-irradiated cholera bacteriophages . U.v . light-induced DNA damage in phages of different morphological and serological groups could be efficiently photoreactivated . Host cell reactivation of irradiated phages of different groups was different on the same indicator host . Phage phi 149 was the most sensitive, and phi 138 the most resistant to u.v . irradiation . While phi 138 showed appreciable host cell reactivation, this was minimal for phi 149 . Attempts to demonstrate Weigle reactivation of u.v.-irradiated cholera phages were not successful, although u.v.-induced filamentation of host cells was observed. Infect Immun, 1983 Aug, 41(2), 735 - 41 Successful colonization and immunization of adult rabbits by oral inoculation with Vibrio cholerae O1; Cray WC Jr et al.; Adult rabbits were inoculated orally (or duodenally) with virulent Vibrio cholerae O1 . Jejunal colonization occurred only when hypoperistalsis was induced at the time of inoculation by tincture of opium given intraperitoneally (or by temporary ileal obstruction) . For oral inoculation, prior neutralization of gastric acid was also required . Inoculation with 10(9) V . cholerae caused jejunal colonization for 1 to 2 days and ileal colonization for 5 to 6 days . The extent of small bowel colonization 18 h after oral inoculation was related to inoculum size but also reflected limited multiplication of small inoculum sizes and net death, clearance of large inoculum sizes, or both . Serious diarrhea occurred only in rabbits fed large inoculum sizes, i.e., 10(10) V . cholerae, and then rarely . Rabbits colonized once with 10(10) V . cholerae became highly resistant to recolonization with either the same or opposite serotype . After 18 weeks, these rabbits were still partially protected, whereas twice-colonized rabbits were highly protected . Protection against recolonization appeared to be due, at least partly, to interference with the adherence of V . cholerae to the bowel mucosa, thus allowing rapid removal of V . cholerae when peristalsis resumed . Prior colonization also protected against cholera-like diarrhea in rabbits challenged by the removable intestinal tie-adult rabbit diarrhea technique, the 50% effective dose for severe or lethal diarrhea being increased more than 100-fold, and probably more than 10,000-fold, for challenge with either the homologous or heterologous serotype of V . cholerae . The described rabbit model appears well suited for the study of immunity evoked by enteric colonization with V . cholerae O1. Antibiotiki, 1983 Aug, 28(8), 592 - 5 {Antibiotic sensitivity of bacteria of the family Vibrionaceae isolated from the intestines of swine}; Andrusenko IT et al.; Bacteria of Aeromonas hydrophila subsp., biotype 1 and Aeromonas anaerogenes, biotype 2, as well as Proteolitica of the vibrio pasahumoliticus biotype were isolated from the intestine of pigs with diarrhea . Close antibiotic sensitivity of the isolates was shown . The differences in their sensitivity were not sufficient for defining the taxonomic features. Appl Environ Microbiol, 1983 Aug, 46(2), 425 - 9 Broth medium for enrichment of Vibrio fluvialis from the environment; Nishibuchi M et al.; A medium was designed for the enrichment and enumeration of Vibrio fluvialis from environmental samples . The medium contains 1% peptone plus 4% sodium chloride and 5 micrograms of novobiocin per ml, pH 8.5 . This V . fluvialis enrichment medium (FEM) was tested, in comparison with alkaline peptone (AP), in field samplings . A total of 177 samples (estuarine waters and sediment, sewage, and crabs) collected over a 14-month period were examined with FEM and with AP broth . Results showed that FEM was more effective than AP in detecting V . fluvialis, particularly from water and sewage samples with low salinities (less than 6%) . The best recovery of V . fluvialis occurred when both enrichment media were used simultaneously. J Clin Microbiol, 1983 Aug, 18(2), 400 - 7 Presumptive identification of Vibrio species with H antiserum; Tassin MG et al.; Species in the genus Vibrio exhibit flagellar (H) antigens unique to the species . Thus, species-specific H antiserum could be a valuable reagent with which to screen serologically large numbers of Vibrio isolates . Antisera against V . cholerae, V . fluvialis, V . anguillarum, V . metschnikovii, V . parahaemolyticus, V . alginolyticus, and V . vulnificus H antigens was produced in rabbits by repeated injections of Formalin-preserved whole cells . Anti-O activity and anti-H activity against common H antigens was absorbed from each antiserum, V . fluvialis was shown to possess an H antigen unique to the species and also to share minor H antigens with V . cholerae, V . metschnikovii, and V . anguillarum . V . vulnificus also exhibits a species-unique H antigen . A comprehensive serological screening system based on species-specific H antiserum was developed to identify pathogenic Vibrio species one step beyond primary isolation . Vibrio species were correctly identified with accuracies ranging from 93 to 100% . Some isolates were either nonmotile or poorly so and thus did not flocculate in H antiserum. Chem Biol Interact, 1983 Aug 1, 45(3), 315 - 26 DNA damage, prophage induction and mutation by furazolidone; Chatterjee SN et al.; Ultraviolet absorption data and thermal chromatography through hydroxyapatite (HAP) column revealed that furazolidone treatment of Vibrio cholerae cells produced more than 80% of DNA reversibly bihelical due to the formation of interstrand cross-links and the reaction obeyed a first order relation . Sensitivities of the Escherichia coli strains to the lethal action of the drug were in the order: AB 2480(uvr- rec-) greater than AB 2463(rec-) greater than AB 1886(uvr-) greater than AB 1157(repair proficient) or AB 4401(wild type) . Furazolidone was 'Rec test' positive, produced dose-dependent prophage induction in E . coli cells and also dose-dependent streptomycin-resistance forward mutation in V . cholerae cells . The quantitative aspect and also the mode of furazolidone action on DNA were discussed. Zh Mikrobiol Epidemiol Immunobiol, 1983 Aug, (8), 45 - 6 {Serovars of nonagglutinating vibrios isolated on the territory of the Uzbek SSR and their role in the development of acute intestinal diseases}; Ziiaev ShI et al.; As the result of the serotyping of 1,223 NAG vibrio strains isolated in the Uzbek SSR from the environment and 659 strains isolated from gastroenteritis patients and from vibriocarriers, the serovars of 563 strains (46%) and 420 strains (63%), respectively, were determined . The prevailing serovars were 08, 06, 09, 014, 05, 039, 041, 013, 019 among the strains isolated from the environment and 08, 041, 037, 05, 039, 024, 06 among the strains isolated from humans . The strains belonging to serovars 05, 06, 08, 024, 037, 039, 041 were isolated both from the environment and from humans . The possible role of these strains in the development of acute gastroenteritis and in the formation of vibriocarriership in humans is pointed out. Biochem Biophys Res Commun, 1983 Jul 18, 114(1), 113 - 8 Isolation of Vibrio alginolyticus mutants defective in the respiration-coupled Na+ pump; Tokuda H; When the respiration-coupled Na+ pump functions, V . alginolyticus is able to grow in the presence of an extremely high concentration of proton conductor, carbonylcyanide m-chlorophenylhydrazone . The mutants which became sensitive to the proton conductor were isolated and examined in regard to the Na+ pump activity . Although the activity of a respiration-dependent H+ extrusion by the mutants is comparable to that by the wild type, the Na+ pump activity of the mutants is significantly reduced . Furthermore, NADH oxidase of membranes isolated from the mutants is altered to be independent of Na+ . It is concluded that the mutants have an alteration in the respiratory chain which simultaneously results in a lack of the Na+ pump. Science, 1983 Jul 15, 221(4607), 289 - 91 Synthesis and secretion of the plasmid-coded heat-labile enterotoxin of Escherichia coli in Vibrio cholerae; Neill RJ et al.; Both cholera toxin and heat-labile enterotoxin were made and secreted into culture supernatants by Vibrio cholerae containing the enterotoxin plasmid pCG86 . Several regulatory mutations in V . cholerae that increased or decreased the synthesis of cholera toxin did not affect production of heat-labile enterotoxin . In contrast, a mutation in V . cholerae that interfered with the secretion of cholera toxin also decreased the secretion of heat-labile enterotoxin, indicating that they are processed by a common secretory pathway . Vibrio cholerae should be useful as a model system for analyzing the secretion of true extracellular proteins by Gram-negative bacteria. Am J Trop Med Hyg, 1983 Jul, 32(4), 812 - 7 Persistence of cholera in the United States; Shandera WX et al.; In 1973, 1978, and 1981, cases of cholera were acquired along the Gulf Coast of the United States . The isolates from all of the cases were toxigenic Vibrio cholerae O-group 1, biotype El Tor, serotype Inaba, hemolytic, and of the same phage sensitivity pattern, and all had the same restriction endonuclease pattern by molecular genetic analysis . The strain from one of the two 1981 cases differed from the others in having a small plasmid and a negative Voges-Proskauer reaction . Multiple importations, chronic carriers, and continuous occurrence of undetected cases are unlikely explanations for these findings, which suggest that toxigenic V . cholerae 01 can multiply and persist for years in some environments, making eradication of cholera a formidable task. J Gen Microbiol, 1983 Jul, 129 (Pt 7), 2193 - 7 Multiflagellate variants of Vibrio anguillarum; Chart H; An ultrastructural examination of six strains of Vibrio anguillarum of varying virulence for eels revealed an apparent correlation between pathogenicity and the possession of more than one flagellum . The relationship between V . anguillarum surface appendages and virulence is discussed. Zh Mikrobiol Epidemiol Immunobiol, 1983 Jul, (7), 51 - 5 {Adhesive properties of Vibrio cholerae strains and mutants with varying biological characteristics}; Uraleva VS et al.; V . cholerae strains and mutants devoid of adhesive activity and possessing very high adhesive activity were found in the collection under study . As a rule, cholerigenic strains possessed high or moderate adhesive activity, while in noncholerigenic strains the coefficient of adhesiveness varied over a wide range . The existence of a definite pronounced correlation between lecithinase activity and adhesiveness was established . Hemolysin-producing strains were found to possess high adhesive activity considerably more frequently than nonhemolytic strains. Zh Mikrobiol Epidemiol Immunobiol, 1983 Jul, (7), 92 - 7 {Modification of a method of passive immune hemolysis on a solid medium for detecting the production of thermolabile enterotoxins by Vibrio cholerae and Escherichia coli strains}; Shaginian IA et al.; A modification of the passive immune hemolysis method for the determination of the production of thermolabile enterotoxins by V . cholerae and E . coli is proposed . This modification permits the use of solid culture media . Experiments with cholera enterotoxin have demonstrated that the sensitivity of the modified method is 8-10 times higher than that of the Elek method . Similar results have been obtained with the use of the proposed method in the study of the capacity of different V . cholerae and E . coli strains for producing enterotoxins . The results obtained with the use of this method have been found to correlate with those obtained by means of the skin test and passive immune hemolysis in a liquid medium . We have used the modified method in the study of the production of thermolabile enterotoxin in transconjugants obtained by the hybridization of E . coli strain GA 107 carrying plasmid pCG86 which determines the synthesis of thermolabile and thermostable enterotoxins and E . coli strain K12 C600 R . The results obtained in the study of toxin formation in 99 transconjugants, carried out with the use of the proposed method, the skin test and passive immune hemolysis, have been shown to coincide. Zh Mikrobiol Epidemiol Immunobiol, 1983 Jul, (7), 55 - 9 {Model UF-2 mutant of Staphylococcus aureus 209 P for titrating Vibrio cholera enterotoxin}; Pasternak NA et al.; The identical character of the action of crude V . cholerae enterotoxin on the anaerobic dehydrogenases of the UV-2 mutant of S . aureus 209 p and the surviving culture of Ehrlich's carcinoma has been revealed . The range of this action is linked with the concentration of the toxin and varies from the stimulation of cell dehydrogenases to their complete suppression . The rapid method for the titration of the enterotoxin in the dehydrogenase suppression test with the use of the bacterial model is proposed. J Periodontol, 1983 Jul, 54(7), 420 - 30 The periodontal microflora of juvenile diabetics . Culture, immunofluorescence, and serum antibody studies; Mashimo PA et al.; These studies demonstrate a unique constellation of organisms populating the subgingival area in periodontitis lesions of patients with juvenile or insulin-dependent diabetes mellitus (IDDM) . The cultivable microflora was predominated by Capnocytophaga and anaerobic vibrios in the patients studied . In some patients, Actinobacillus actinomycetemcomitans were also found . This distinguishes the subgingival flora of IDDM patients suffering from periodontitis from that of patients with localized juvenile periodontitis (LJP), and that of adult periodontitis patients . In LJP most patients harbor both A actinomycetemcomitans and Capnocytophaga subgingivally; and in periodontitis lesions from nondiabetic adults, black-pigmented Bacteroides such as B gingivalis or B melaninogenicus subspecies intermedius are often found . Antibiotic susceptibility patterns suggest that penicillin or tetracycline or its analogs such as minocycline may be effective against the predominant cultivable microflora in periodontal lesions of IDDM patients; however, individual patients may harbor flora with significant resistance to these antibiotics. Infect Immun, 1983 Jul, 41(1), 237 - 43 Purification of Vibrio cholerae soluble hemagglutinin and development of enzyme-linked immunosorbent assays for antigen and antibody quantitations; Svennerholm AM et al.; Soluble hemagglutinin (HA) from an El Tor Vibrio cholerae strain (serotype Ogawa) was purified by means of a sequence of salt precipitation, gel filtration, and agarose electrophoresis . The purified material, which gave a single precipitation line in immunodiffusion tests with homologous antiserum, showed immunological identity reactions in double diffusion-in-gel with soluble HA produced by various classical and El Tor strains of different serotypes . Purified HA was used for development of an enzyme-linked immunosorbent assay for titration of specific antibodies against soluble HA and for quantitation of this antigen . Rabbit anti-HA serum reacted in high titer with the soluble HA coated on polystyrene microtiter plates, whereas antiserum against cholera toxin, lipopolysaccharide, or whole washed V . cholerae showed little or no reactivity . In inhibition tests as little as 2.5 ng of soluble HA could be detected with the enzyme-linked immunosorbent assay . Culture supernatants of different El Tor as well as classical V . cholerae strains all completely inhibited the binding of anti-HA antibody to solid-phase-bound homologous antigen, but the amounts of HA produced by individual strains varied at least 1,000-fold . Only 2 of 10 paired acute- and convalescent-phase sera from Bangladeshi cholera patients showed significant titer increases against soluble HA in parallel titrations. Biochim Biophys Acta, 1983 Jun 23, 731(3), 487 - 90 Differential effect of neuraminidase-treatment on the surface charge-associated properties of rat reticulocytes and erythrocytes . Studies by partitioning in two-polymer aqueous phases; Walter H et al.; Rat reticulocytes undergo charge-associated surface changes, detectable by cell partitioning in charged dextran-poly(ethylene glycol) aqueous phase systems, as they become mature erythrocytes . Young reticulocytes have a lower partition coefficient, i.e., quantity of cells in the top phase as a percentage of total cells added, than do mature erythrocytes . Sialic acid is the main charge-bearing group on red blood cells and, in the case of the rat, most of the sialic acid can be removed by treatment of the cells with neuraminidase (Vibrio cholerae) . By combining isotopic 59Fe-labeling of reticulocytes with countercurrent distribution of the entire red blood cell population in charged dextran-poly(ethylene glycol) aqueous phases we have now studied the relative effect of neuraminidase-treatment on rat reticulocytes and mature erythrocytes . It was found that neuraminidase-treatment (a) does not eliminate surface differences, detectable by partitioning, between rat reticulocytes and erythrocytes and (b) reduces the partition coefficient of mature erythrocytes to a greater extent than the partition coefficient of reticulocytes indicating a differential effect of this enzyme on the two cell populations. N Engl J Med, 1983 Jun 9, 308(23), 1389 - 92 Protection against cholera in breast-fed children by antibodies in breast milk; Glass RI et al.; We performed a prospective study to examine whether the IgA antibodies against cholera that are present in breast milk protect breast-fed infants and children against colonization with Vibrio cholerae 01 and disease . Among families of patients with cholera, we collected breast milk from mothers who had not had diarrhea in the previous week and monitored them and their breast-fed children for cholera colonization and diarrhea for 10 days . Breast milk was assayed for IgA antibodies to cholera toxin and lipopolysaccharide . Ninety-three mother--child pairs were studied; 30 infants became colonized with V . cholerae 01 and disease developed in 19 . There were no differences between the antibody levels in milk fed to children who became colonized and in milk fed to children who did not . However, among the children who became colonized, those who had diarrhea drank breast milk containing significantly lower levels of both kinds of cholera antibodies than were present in the milk consumed by children who had no symptoms . We conclude that breast-milk antibodies against cholera do not appear to protect children from colonization with V . cholerae 01 but do protect against disease in those who are colonized. J Clin Microbiol, 1983 Jun, 17(6), 1102 - 13 Numerical taxonomy of Vibrio cholerae and related species isolated from areas that are endemic and nonendemic for cholera; McNicol LA et al.; A total of 165 strains of vibrios isolated from clinical and environmental sources in the United States, India, and Bangladesh, 11 reference cultures, and 4 duplicated cultures were compared in a numerical taxonomic study using 83 unit characters . Similarity between strains was computed by using the simple matching coefficient and the Jaccard coefficient . Strains were clustered by unweighted average linkage and single linkage algorithms . All methods gave similar cluster compositions . The estimated probability of error in the study was obtained from a comparison of the results of duplicated strains and was within acceptable limits . A total of 174 of the 180 organisms studied were divided into eight major clusters . Two clusters were identified as Vibrio cholerae, one as Vibrio mimicus, one as Vibrio parahaemolyticus, three as Vibrio species, and one as Aeromonas hydrophila . The V . mimicus cluster could be further divided into two subclusters, and the major V . cholerae group could be split into seven minor subclusters . Phenotypic traits routinely used to identify clinical isolates of V . cholerae can be used to identify environmental V . cholerae isolates . No distinction was found between strains of V . cholerae isolated from regions endemic for cholera and strains from nonendemic regions. J Pharmacol Exp Ther, 1983 Jun, 225(3), 595 - 8 Antibiotic efficacy against Vibrio vulnificus in the mouse: superiority of tetracycline; Bowdre JH et al.; Seven antimicrobial agents, all effective against Vibrio vulnificus in vitro, were compared for in vivo efficacy in mice experimentally infected with V . vulnificus strain B3547 . Mice were injected s.c . with 1 X 10(8) cells, and i.p . injection of antimicrobials was begun 1.5 hr later when mice were bacteremic and had edematous lesions at the injection site . The study was done in two phases . Phase I was a dose-ranging experiment, using single injections within the range (on a body weight-adjusted basis) clinically useful in humans . Of 12 mice treated with tetracycline (4 mg/kg), 12 survived at 24 hr, compared to 0 of 21 saline-treated controls, 3 of 10 given ampicillin (32 mg/kg) and 2 of 3 given cefotaxime (20 mg/kg) . There were no survivors at 24 hr in groups of 5 to 10 mice treated with cefazolin (32 mg/kg), carbenicillin (80 mg/kg), erythromycin (8 mg/kg) or gentamicin (8 mg/kg) . Phase II was designed to simulate clinical conditions using multiple