Biochim Biophys Acta, 1985 Dec 16, 810(3), 332 - 9 Phosphorylation and phosphate-ATP exchange catalyzed by the ATP synthase isolated from Wolinella succinogenes; Bokranz M et al.; The ATP synthase, isolated from Wolinella (formerly Vibrio) succinogenes could be fully incorporated into liposomes without significant cleavage of the enzyme or loss of activity . These proteoliposomes, but not the isolated enzyme, catalyzed phosphate-ATP exchange and the phosphorylation of ADP which was driven by an artificially imposed delta mu H across the liposomal membrane . Phosphorylation driven by light was catalyzed by proteoliposomes containing also bacteriorhodopsin . The three activities were similarly sensitive to protonophores or dicyclohexylcarbodiimide . This sensitivity was similar to that of the electron-transport-driven phosphorylation catalyzed by bacterial membrane vesicles . With a delta mu H value 280 mV to drive phosphorylation the turnover number of the enzyme was in the same order of magnitude as that measured in the electron-transport-driven phosphorylation catalyzed by the bacterial membrane . When the delta mu H was below 150 mV, the phosphorylation activity of the incorporated enzyme was two orders of magnitude slower, and was about as fast as light-driven phosphorylation or as the exchange reaction. Br Med J (Clin Res Ed), 1985 Dec 7, 291(6509), 1601 - 5 Clinical trial of berberine in acute watery diarrhoea; Khin-Maung-U et al.; Four hundred adults presenting with acute watery diarrhoea were entered into a randomised, placebo controlled, double blind clinical trial of berberine, tetracycline, and tetracycline and berberine to study the antisecretory and vibriostatic effects of berberine . Of 185 patients with cholera, those given tetracycline or tetracycline and berberine had considerably reduced volume and frequency of diarrhoeal stools, duration of diarrhoea, and volumes of required intravenous and oral rehydration fluid . Berberine did not produce an antisecretory effect . Analysis by factorial design equations, however, showed a reduction in diarrhoeal stools by one litre and a reduction in cyclic adenosine monophosphate concentrations in stools by 77% in the groups given berberine . Considerably fewer patients given tetracycline or tetracycline and berberine excreted vibrios in stools after 24 hours than those given berberine alone . Neither tetracycline nor berberine had any benefit over placebo in 215 patients with non-cholera diarrhoea. Life Sci, 1985 Dec 2, 37(22), 2059 - 65 Attenuation of inhibitory processes in the central nervous system by tetanus toxin: an in vitro study on rat hippocampal slices; Wieraszko A; The influence of tetanus toxin on the efficiency of recurrent inhibition in the rat hippocampal slice was tested . The efficiency of the recurrent inhibition diminished in a dose-dependent manner following incubation of the slices with tetanus toxin . The effect was not observed in the slices preincubated for 3 hours with neuraminidase from Vibrio cholerae . This treatment reduces markedly the level of polysialogangliosides (receptor for tetanus toxin) . It is concluded that tetanus toxin influences the efficiency of some inhibitory synapses in the central nervous system and that a certain level of polysialogangliosides is necessary for tetanus toxin to exert its action. FEBS Lett, 1985 Dec 2, 193(2), 250 - 4 Amino acid sequence of heat-stable enterotoxin produced by Vibrio cholerae non-01; Takao T et al.; The amino acid sequence of heat-stable enterotoxin, produced by Vibrio cholerae non-01 and isolated from its culture supernatant, was determined by both Edman degradation of native and reductively carboxymethylated enterotoxin and also a combination of fast atom bombardment mass spectrometry and carboxypeptidase Y digestion of native enterotoxin to be as follows: Ile-Asp-Cys-Cys-Glu-Ile-Cys-Cys-Asn-Pro-Ala-Cys-Phe-Gly-Cys-Leu-Asn . This sequence is very similar, but not identical, to those of heat-stable enterotoxins produced by enterotoxigenic Escherichia coli and Yersinia enterocolitica. Appl Environ Microbiol, 1985 Dec, 50(6), 1548 - 9 Uptake and clearance of Vibrio vulnificus from Gulf coast oysters (Crassostrea virginica); Kelly MT et al.; Oysters collected in late winter, when they were free of Vibrio vulnificus, were exposed in the organism in the laboratory . The oysters effectively concentrated the bacteria from seawater, but when the inoculum was removed, the bacteria were rapidly cleared from the oyster tissues . These results suggest that V . vulnificus may be found in oysters as a result of filtration of the bacteria from seawater rather than active multiplication of the bacteria in the oysters. Appl Environ Microbiol, 1985 Dec, 50(6), 1388 - 94 Characterization and distribution of Vibrio alginolyticus and Vibrio parahaemolyticus isolated in Indonesia; Molitoris E et al.; Previous studies have shown that Vibrio alginolyticus and Vibrio parahaemolyticus can be isolated from similar types of marine samples . In this report, the results of an examination of 567 V . alginolyticus and V . parahaemolyticus strains, isolated from seawater in Jakarta Bay and from more than 30 types of seafood from markets in Jakarta, Indonesia, are presented . Most isolates were from mackerel, shrimp, or squid . Numerical taxonomic analyses clustered 337 isolates and three V . alginolyticus reference strains at S greater than or equal to 80% . These strains produced acid from sucrose, but only approximately 80% produced acetoin or grew in the presence of 10% NaCl . The frequency of occurrence of V . alginolyticus in seawater samples ranged from 0% (in February and March 1972) to 100% (in September and December 1972) and was highest in seafood samples from August to December 1972 . A second cluster of 230 isolates and seven V . parahaemolyticus reference strains was observed at S greater than or equal to 82% . These strains did not produce acetoin or acid from sucrose, and approximately 20% grew in the presence of 10% NaCl . V . parahaemolyticus was detected in seawater samples each month, with the highest frequency of occurrence (83.3%) in May 1972 . Twenty-nine K antigen serotypes were demonstrated in V . parahaemolyticus isolates, and another 40% were untypable . The modal antibiotic resistance pattern for each species included five drugs . Only 12% of the V . parahaemolyticus strains were Kanagawa positive, and 10% elicited fluid accumulation in ligated rabbit ileal loops . All of the 7 V . alginolyticus strains and 94 (70%) of the V . parahaemolyticus strains tested killed mice when inoculated intraperitoneally.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1985 Dec, 50(3), 922 - 4 Cloning of the cytotoxin-hemolysin gene of Vibrio vulnificus; Wright AC et al.; Genes encoding the cytotoxin-hemolysin of Vibrio vulnificus were cloned in Escherichia coli by using the lytic cloning vector, lambda 1059 . Subcloning in plasmid pBR325 resulted in the isolation of a 3.2-kilobase DNA fragment containing the cytotoxin gene . By using this fragment as a DNA probe, homologous gene sequences were detected in all 54 V . vulnificus strains studied; homologous sequences were present in none of 96 isolates from 29 other bacterial species. Infect Immun, 1985 Dec, 50(3), 813 - 6 Role of cholera toxin in enteric colonization by Vibrio cholerae O1 in rabbits; Pierce NF et al.; The role of cholera toxin (CT) in mucosal colonization by Vibrio cholerae O1 was studied in rabbits by using toxinogenic V . cholerae and nontoxinogenic (A-B+ or A-B-) recombinant mutants derived from them . After oral inoculation, toxinogenic strains colonized intestinal mucosa significantly more efficiently than did either A-B- or A-B+ mutants; average colonization was increased 1.5- to 30-fold with toxinogenic strains, depending on the inoculum used and the portion of intestine studied . Additionally, colonization by an A-B- mutant was increased to the levels of its toxinogenic parent by coadministration of CT with the inoculum . We conclude that CT contributes significantly to mucosal colonization by V . cholerae and that this effect is not due to an interaction of the CT B subunit with its mucosal receptor . The possibility that this effect contributes to the in vivo selection of hypertoxinogenic variants of V . cholerae is considered. J Clin Microbiol, 1985 Dec, 22(6), 1011 - 3 Modified taurocholate-tellurite-gelatin agar for improved differentiation of Vibrio species; O'Brien M et al.; A total of 78 strains, representing 21 Vibrio species, were examined by using taurocholate-tellurite-gelatin agar (TTGA) medium and modified TTGA medium containing 4-methylumbelliferyl-beta-D-galactoside (150 micrograms/ml) . Modified TTGA medium allowed for simple and direct detection of beta-D-galactosidase (beta-gal) activity . This feature, in conjunction with other differential characteristics of TTGA medium, gave improved differentiation of the vibrios tested . The modified TTGA medium allowed for easy differentiation of Vibrio cholerae (beta-gal+) from Vibrio parahaemolyticus (beta-gal-) . The 4-methylumbelliferyl-beta-D-galactoside substrate is inexpensive and very stable . Incorporation into the agar did not affect the performance of TTGA as a differential medium . The assay for beta-gal activity with this substrate was specific and sensitive. Zh Mikrobiol Epidemiol Immunobiol, 1985 Dec, (12), 30 - 5 {Preparation and use of magnetic sorbents for studying microorganism antigens}; Pushkar' VG et al.; In this work simple techniques for obtaining polyacrylamide sorbents with magnetic properties are described . These techniques have permitted obtaining block and microgranulated sorbents with the immobilization of antibodies from plague antiserum in the cellular gel structure for the specific sorption of killed and live Yersinia pestis cells and their first fraction; pig brain gangliosides have also been incorporated into the gel structure with a view to the sorption of cholera toxin from the filtrate of Vibrio cholerae culture . The magnetic properties of sorbents, obtained by the copolymerization of powdered magnetic ferric oxides in gel, have made it possible to increase the effectiveness of specific sorption due to mixing and rapid separation in different magnetic fields, as well as to facilitate and accelerate manipulations with the sorbent at all stages . The capacity of different types of sorbents and the time of sorption have been determined. Anal Biochem, 1985 Nov 15, 151(1), 137 - 41 Separation of bacterial luciferase from oxidoreductases by affinity chromatography; Tsai TS; The NADH-dependent and NADPH-dependent oxidoreductase activities associated with bacterial luciferase in Vibrio (Beneckea) harveyi can simultaneously be removed from purified luciferase through a Blue Sepharose CL-6B column . The result achieved with this one-step affinity chromatography is similar to that obtained with two sequential "reverse-affinity" chromatographies. Eur J Biochem, 1985 Nov 15, 153(1), 89 - 94 Bacteriophage CP-T1 of Vibrio cholerae . Identification of the cell surface receptor; Guidolin A et al.; The attachment site on the cell surface of Vibrio cholerae for the bacteriophage CP-T1 has been determined . Purified lipopolysaccharide from the Inaba and Ogawa serotypes, and of both the Classical and El Tor biotype of strains of V . cholerae show equal phage-inactivating capacities . Lipopolysaccharide extracted from a CP-T1-resistant mutant has no phage-inactivating capacity . Such mutants lack O-antigen as demonstrated by bactericidal assays utilizing a monoclonal antibody directed against O-antigen side chain of V . cholerae lipopolysaccharide . Radiolabelling of lipopolysaccharide with 33P and analysis by sodium dodecyl sulfate/polyacrylamide gel electrophoresis also revealed the absence of O-antigen in phage-resistant strains . A number of V . cholerae typing phage show cross-resistance with phage CP-T1. J Virol, 1985 Nov, 56(2), 356 - 64 Effect of neuraminidase treatment of cells and effect of soluble glycoproteins on type 3 reovirus attachment to murine L cells; Gentsch JR et al.; The effect of pretreatment of murine L cells with bacterial neuraminidases on type 3 reovirus attachment was examined . We observed that such treatments resulted in a 60 to 80% decrease of subsequent attachment of 35S-labeled type 3 reovirus in a time- and dose-dependent manner . This result was specific for removal of cell surface sialic acid residues since the specific neuraminidase inhibitor 2-deoxy-2,3-dehydro-n-acetyl neuraminic acid completely prevented the observed effect . Although the total amount of radiolabeled virus bound to neuraminidase-treated cells was greatly reduced, unlabeled reovirus competed only slightly less efficiently for the attachment of 35S-labeled reovirus to neuraminidase-treated versus mock-treated L cells, suggesting that the specificity of the virus interaction with cellular receptor sites was only slightly diminished . Saturation experiments with mock-treated cells or with cells treated with Vibrio cholerae or with V . cholerae plus Arthrobacter ureafaciens neuraminidases indicated that the number of specific cellular receptor sites for type 3 reovirus were reduced by about 47% . We determined that under the neuraminidase digestion conditions used in this experiment we were able to remove a maximum 75% of the total N-acetylneuraminic acid of L cells . Our results also demonstrated that glycoproteins bearing a large amount of sialic acid containing oligosaccharides as well as purified N-acetylneuraminic acid, N-glycolylneuraminic acid, and N-acetylneuraminyl lactose were inhibitors of attachment, while proteins containing no sialic acid or negligible amounts of sialic acid did not inhibit attachment . High concentrations of various monosaccharides and lactose had no effect on reovirus attachment, in agreement with the recent results of Armstrong and his collaborators (Armstrong et al., Virology, 138:37-48, 1984) . These data are also supported by the observation that gangliosides are inhibitors of viral attachment (Armstrong et al., Virology, 138:37-48, 1984) . Taken together, our results suggest that cell surface sialic acid-containing glycoconjugates are involved in type 3 reovirus binding to murine L cells. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1985 Nov, 260(3), 311 - 8 Enterotoxigenicity of clinical isolates of non-O1 Vibrio cholerae; Shehabi AA et al.; Whole cultures, but not culture supernatant fluids, of 21 isolates of non-O1 V . cholerae from patients with diarrhea were shown to induce positive fluid accumulation in infant mice . CHO cell assays demonstrated the elaboration of heat-labile cytotonic, cytotoxic or both factors from most isolates when grown under optimal culture conditions . These factors were not neutralized by anti-cholera toxin serum . Also genetic studies performed on 9 vibrio isolates using a DNA hybridization probe failed to detect gene sequences homologous with cholera toxin . ELISA assays recognized six isolates which produced a cell-associated substance which immunologically cross-reacted with cholera toxin . Enzymatic profiles of the vibrio isolates did not correlate with the production of any toxic factor . The results indicate that mainly heat-labile and cell-associated cytotonic and cytotoxic factors appear to influence the enterotoxigenic potential of this heterogenous group of non-O1 vibrios. Diagn Microbiol Infect Dis, 1985 Nov, 3(6), 461 - 8 Identification of Vibrio vulnificus by indirect immunofluorescence; Gray LD et al.; A rapid, sensitive, and specific indirect immunofluorescence (IIF) procedure is described for identifying Vibrio vulnificus . Reference antisera were prepared by vaccinating rabbits with surface antigen preparations of V . vulnificus, and the antisera were examined for the ability to react with and serologically group 85 isolates of V . vulnificus grown in heart infusion broth, and to detect V . vulnificus in tissue specimens from mice experimentally infected with a virulent isolate of the bacterium . The antisera detected 100% of the V . vulnificus isolates examined and gave false-positive results in approximately 0.9% of 445 IIF tests performed with non-V . vulnificus clinical isolates . V . vulnificus also was detected in frozen tissue sections from infected mice; however, the most easily observed positive results were obtained by examining V . vulnificus from lesion specimens and blood cultured briefly in heart infusion broth . The bacteria in 2-hr-old cultures of local lesions fluoresced brilliantly and were easily detectable . The IIF procedure could be of value in rapidly diagnosing fulminating and potentially fatal human disease caused by V . vulnificus. J Clin Microbiol, 1985 Nov, 22(5), 868 - 9 Comparison of the modified Elek test and Wagatsuma agar for determination of the Kanagawa phenomenon of Vibrio parahaemolyticus; Nair GB et al.; The modified Elek test and Wagatsuma agar were compared for their ability to detect the Kanagawa activity of 142 strains of Vibrio parahaemolyticus . The performance of the modified Elek test was on a par with that of the Wagatsuma agar as far as positivity was concerned, and the test was far superior to Wagatsuma agar in eliminating doubtful results . The results of the modified Elek test were not unduly influenced by the different types of agar used. Infect Immun, 1985 Nov, 50(2), 534 - 40 Production and partial characterization of an elastolytic protease of Vibrio vulnificus; Kothary MH et al.; Conditions are described for the production of large amounts of an extracellular elastolytic protease by Vibrio vulnificus . The yield of enzyme was maximal during the late exponential growth phase and was stable during the stationary growth phase in a medium composed of 2% Proteose Peptone and 1.5% NaCl . The protease has a molecular weight of ca . 50,500 (estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis), an isoelectric point of ca . 5.8, and a pH optimum range against azocasein and elastin of pH 7 to 8 . The caseinolytic and elastase activities in protease preparations partially purified by ammonium sulfate precipitation were inseparable by gel filtration, hydrophobic interaction chromatography, and isoelectric focusing . Both activities were deleteriously affected by heat, low pH, heavy-metal ions, chelating agents, reducing agents, sodium cyanide, N-bromosuccinimide, alpha-2-macroglobulin, and phosphoramidon, but were unaffected by various trypsin inhibitors, chymostatin, aprotinin, leupeptin, pepstatin A, phenylmethylsulfonyl fluoride, and N-ethylmaleimide. Ann Inst Pasteur Microbiol, 1985 Nov-Dec, 136B(3), 265 - 73 Genetic basis of trimethoprim and O/129 resistance in Vibrio cholerae; Gerbaud G et al.; Because of its important consequences on prophylaxis and therapy of cholera and on bacterial identification, we have studied the genetic basis of cross-resistance to trimethoprim and O/129 of strains of Vibrio cholerae O1 independently isolated in Africa . Two classes of bacteria were found . In the first class, the strains were also resistant to ampicillin and kanamycin and to high levels of streptomycin by synthesis of a 3"- or 6-aminoglycoside phosphotransferase . The strains hybridized weakly with a Tn7 probe and all the resistance characters were transferable en bloc to Escherichia coli . The second class included strains which, in addition to trimethoprim and O/129, were resistant to moderate levels of streptomycin and spectinomycin by production of a 3",9-aminoglycoside-aminocyclitol adenylyltransferase . The resistance characters were not self-transferable to E . coli and the host strain hybridized strongly with Tn7 . It therefore appears, that both plasmids and transposons are responsible for the dissemination of resistance to trimethoprim and O/129 in Vibrio. Int J Cancer, 1985 Oct 15, 36(4), 461 - 6 Expression of cell surface glycoproteins in human melanoma cell lines with different tumorigenic properties; Berthier-Vergnes O et al.; Human malignant melanoma cell lines characterized by either a high or a low ability to grow subcutaneously in athymic nude mice have been examined for their cell-surface glycoproteins . Striking differences were demonstrated between these 2 groups . Cells from lines of low tumorigenicity (LT group) displayed twice as much Vibrio cholerae neuraminidase and galactose oxidase accessible glycoproteins as cells from lines of high tumorigenicity (HT group) and each group of cell lines could be characterized by specific glycoprotein profiles . LT and HT group cells displayed similar amounts of periodate accessible glycoproteins, but sialoglycoprotein profiles were characteristic for each group of cell lines . Furthermore, whereas 87% of the sialic acid released by V . cholerae neuraminidase came from cell surface glycoproteins in HT group cells, only 53-55% of the released sialic acid came from surface glycoproteins in LT group cells . These results suggest that human melanoma cell lines exhibiting different tumorigenicity in nude mice can also be characterized by differences in composition and organization within the plasma membranes of their cell-surface sialoglycoproteins. Brain Res, 1985 Oct 14, 345(1), 159 - 64 The role of monosialoganglioside GM1 in the synaptic plasticity: in vitro study on rat hippocampal slices; Wieraszko A et al.; Rat hippocampal slices were incubated with neuraminidase from Vibrio Cholerae . This enzyme liberates sialic acid from polysialogangliosides converting them into monosialoganglioside GM1 . Thus, the tissue is enriched in GM1 content . Another set of slices was incubated with GM1 itself . Both treatments increased the magnitude of potentiation of synaptic response recorded from pyramidal cell layer following high frequency stimulation of Schaffer collateral-commissural fibers . It is concluded that enrichment of synaptic membranes in GM1 enhances the ability of these nerve endings to be potentiated. Zh Mikrobiol Epidemiol Immunobiol, 1985 Oct, (10), 8 - 11 {Cholera and NAG vibrio utilization of different C-containing substrates in Hiss media and a medium with a limited mineral content}; Voronezhskaia LG; A total of 55 V . cholerae strains and 175 NAG vibrio strains were studied with a view to establish their capacity for utilizing citrate in Simmons citrate agar or for growing in it in the absence of the source of carbon . The strains were divided into 3 groups, each containing approximately an equal number of cholera and NAG vibrios irrespective of their origin or serovars . None of 50 signs used in this investigation permitted the reliable differentiation of the cholera and NAG vibrio groups due to considerable differences between the strains within each group . The use of Hiss medium with starch instead of Kodam medium is proposed for the determination of the diastatic activity of cholera and NAG vibrios. Zh Mikrobiol Epidemiol Immunobiol, 1985 Oct, (10), 45 - 51 {Cholera morbidity problems in 1 of the departments of the Democratic and Popular Republic of Algeria}; Briko NI et al.; A total of 1,078 cases of bacteriologically confirmed cholera were analyzed at the period of 1979-1983 . In 1981 Vibrio eltor, serotype Inaba, replaced V . cholerae, serotype Ogawa, and became the prevailing infective agent . Every year young children and persons over 50 years of age were most actively involved into the epidemic process . The peak of seasonal morbidity was observed in September-October . The appearance of the foci of infection in families was found to be slightly pronounced in cholera . 85.3% of the families had only a single case of cholera . The cases of cholera with the fatal termination of the disease were registered mostly at the beginning of the seasonal rise of morbidity and at its peak. Zh Mikrobiol Epidemiol Immunobiol, 1985 Oct, (10), 11 - 5 {Determination of the degree of similarity of cholera and NAG vibrios}; Voronezhskaia LG; The numeric taxonomy taking into account 80 signs has demonstrated the similarity of NAG and cholera vibrios, the average similarity coefficient exceeding 80 % . Among NAG vibrios, 53 % of the strains have been found to deviate from the central strain of V . cholerae mainly with regard to their utilization of the sources of carbon . The use of the citrate sign for the study of the biological properties of NAG vibrios is proposed. J Clin Microbiol, 1985 Oct, 22(4), 572 - 5 Isolation of non-O1 Vibrio cholerae associated with enteric disease of herbivores in western Colorado; Rhodes JB et al.; Non-O1 Vibrio cholerae was isolated from a horse (Equus caballus), a lamb (genus Ovis), and two American buffalo (Bison bison) suffering from enteric disease in the western part of Colorado . In 1981, a foal died of apparent respiratory failure . Necropsy findings included heart failure and gastroenteritis . V . cholerae serovar 347 (Smith) was isolated from the colon of this animal . V . cholerae serovar 27 (Smith) was isolated in 1983 from the intestine of a feedlot lamb suffering from pneumonia and severe watery diarrhea . In 1984, an enteric disease occurred in a herd of American bison . The sick animals were depressed and separated from the herd, dying in about 3 days . Of approximately 100 adult bison, 7 died . Necropsy of one animal revealed that gross lesions were limited to the gastrointestinal tract . V . cholerae serovar 27 (Smith) was isolated from the abomasum, duodenum, and colon of this animal . A swab specimen from the intestine of another dead bison also yielded V . cholerae serovar 27 (Smith). J Gen Microbiol, 1985 Oct, 131 ( Pt 10), 2595 - 600 Absence of error-prone repair in a Vibrio species; Thomson JA et al.; The effect of various DNA-damaging agents on a Vibrio species was investigated . The organism was readily mutable by N-methyl-N'-nitro-N-nitrosoguanidine and mitomycin C but not by UV light . No Weigle reactivation of UV-irradiated alpha 3a phage was detected . These results suggest that an error-prone repair mechanism is lacking in this species. J Hyg (Lond), 1985 Oct, 95(2), 299 - 307 Survival of Kanagawa-positive strains of Vibrio parahaemolyticus in a brackish-water area; Kumazawa NH et al.; Vibrio parahaemolyticus was observed to overwinter in sediments and to be present in considerable numbers in sediments and Clithon retropictus (gastropod mollusc) during summer months at a brackish-water area along Hashizu Creek in Japan . The highest level of the organisms was 9.3 X 10(6) and 2.3 X 10(7)/100 g in sediments and C . retropictus respectively . Production of Kanagawa haemolysin was detected in approximately 12% and 20% of strains isolated from sediments and C . retropictus respectively at two stations in Hashizu Creek but were not detected at the other three stations . Two haemolysin-producing strains were isolated from water samples but none were isolated from Corbicula japonica (bivalve mollusc) . These findings suggest that haemolysin producers are preserved principally in sediments and some shellfish in the brackish-water areas with restricted salinity conditions. J Gen Microbiol, 1985 Oct, 131 ( Pt 10), 2653 - 7 Regulation of toxin biosynthesis by plasmids in Vibrio cholerae; Khan AA et al.; Vibrio cholerae strain 569B Inaba harbouring P plasmid produced less toxin than the parent strain . To examine the effect of plasmid loss on toxin production, temperature-sensitive (ts) mutants of P, unable to replicate at 42 degrees C, were isolated . One ts plasmid was unstable at 42 degrees C and its loss yielded a cured strain that resumed a normal level of toxin biosynthesis characteristic of the plasmid-free parent strain . Toxin production was again suppressed in the cured strain after reacquisition of P plasmid . This suggested a role for plasmid-borne genes in the regulation of toxin biosynthesis . A mutant of strain 569B Inaba that produced mutant toxin was isolated by transfer of P and V plasmids . The mutant toxin was similar to choleragenoid because it did not give rise to symptoms of cholera but induced antitoxin immunity in rabbits. Surg Gynecol Obstet, 1985 Oct, 161(4), 357 - 61 Necrotizing fasciitis; Pessa ME et al.; Necrotizing infections of soft tissues are rapidly progressive infections accompanied by a high mortality . Clinical presentation involves fever, cellulitis, edema, crepitus, bullae, necrosis and sepsis . Operative findings include fascial and subcutaneous tissue necrosis with or without myonecrosis . The treatment is prompt surgical debridement . Delay in treatment results in decreased survival time . The mortality in our study was 30 per cent (20 of 33) . The time from recognition of an infection by the patient or physician until operative debridement averaged three and one-half days for survivors compared with 11.7 days for nonsurvivors . These infections frequently occur in compromised hosts and the mortality is greatly increased in these patients . Patients with diabetes had a mortality of 63 per cent (five of eight) . The mortality for infections of the abdomen (44 per cent) and perineum (38 per cent) is greater than for the extremities (18 per cent) . The bacteriologic findings of these infections involved three combinations of organisms . We had 30 mixed infections involving two or more organisms . In addition, three patients had identical signs and symptoms caused by infection with a single organism--Vibrio species . These patients all had some type of contact with a marine environment as a predisposing cause . We also applied both the APACHE and SIS systems to these patients to evaluate the severity of the illness . Nonsurvivors presented with a mean SIS score of 8.64 compared with survivors with 3.82 . Initial scores with the APACHE system for nonsurvivors was 12.0 compared with 2.14 for survivors . In both systems, scores rapidly increased at three and seven days in nonsurvivors as compared with a rapid decline in the survivors . We suggest that the best descriptive system is to simply identify the organisms and tissues involved. J Natl Cancer Inst, 1985 Oct, 75(4), 605 - 11 Organization and neuraminidase susceptibility of sialic acid residues in human melanoma cell lines with different heterotransplantabilities in nude mice; Berthier-Vergnes O et al.; Quantitative analyses of sialic acid residues expressed at the surface of human melanoma cells have been performed on 6 cell lines differing in their ability to grow subcutaneously in nude mice . Whereas 3 of these cell lines showed low heterotransplantability (LT), 3 other cell lines showed high heterotransplantability (HT) . It was found by several methodologic approaches that the 6 human melanoma cell lines varied significantly in their amount of sialic acid susceptible to Vibrio cholerae neuraminidase, but had similar amounts of total sialic acid residues . Cells in the LT group exhibited twice as much cell surface sialic acid residue susceptible to this enzyme as cells in the HT group . Specific fluorescent labeling of external cell surface sialic acid residues showed that the LT cells present a patch-like distribution of the label, whereas the HT cells are characterized by a more homogeneous distribution of the label . Thus the human melanoma cell lines could be distinguished not only by their heterotransplantability in nude mice but also by membrane properties, such as the topographic organization of their cell surface sialic acid residues. J Dent Res, 1985 Oct, 64(10), 1233 - 44 Antibiotic susceptibility of anaerobic bacteria from the human oral cavity; Baker PJ et al.; Anaerobic, agar-dilution, minimal inhibitory concentrations (MICs) of 18 antibiotics are given for the numerically important bacterial groups from the human oral cavity . Strains are divided into susceptibility categories using the guidelines for interpretation of MICs suggested by the National Committee for Clinical Laboratory Standards . These guidelines are based on data on antibiotic concentrations attainable in serum following various dosage regimens . MICs are also compared with attainable gingival fluid levels where these are known . The highest percentages of strains were susceptible to tetracycline, with 89% of the 139 strains tested susceptible to serum levels and 97% conditionally susceptible to attainable gingival fluid levels . Ninety-eight percent of strains were conditionally susceptible to attainable gingival fluid levels of minocycline, but many strains, including Actinobacillus actinomycetemcomitans, were only moderately susceptible to attainable serum levels of this tetracycline analogue . Carbenicillin was effective against most groups of organisms, with the important exception of A . actinomycetemcomitans, at serum levels attainable with oral formulations of carbenicillin . Only 2% of the total strains tested were resistant to penicillin, while 33% of strains were categorized as moderately susceptible . Clindamycin was active against many strains of Gram-negative bacteria but was not active against A . actinomycetemcomitans, some Bacteroides, Eikenella corrodens, or the anaerobic vibrios . Metronidazole was active against A . actinomycetemcomitans, all five groups of oral Bacteroides tested, and against Capnocytophaga species . Chloramphenicol was active against A . actinomycetemcomitans, but not against most of the other groups of oral organisms . Nearly all groups contained strains non-susceptible to serum levels attainable with the usual doses of erythromycin, spiramycin, vancomycin, kanamycin, neomycin, streptomycin, doxycycline, oxytetracycline, or chlortetracycline; several strains were resistant to maximum attainable serum levels of each of these antibiotics except doxycycline. Microbiologica, 1985 Oct, 8(4), 347 - 53 Tryptophanase activity in different toxigenic and nontoxigenic strains of Vibrio cholerae: effect of glucose; Chakraborti MK et al.; Tryptophanase activity was measured in eight different toxigenic and nontoxigenic strains of Vibrio cholerae (V . cholerae) in presence and absence of inducer tryptophan (2 mM) . Stimulation of enzyme activity was observed in both toxigenic and nontoxigenic strains of V . cholerae in presence of inducer . Tryptophanase activity remained much higher in toxigenic strains than that in nontoxigenic strains . Low levels of enzyme activity in nontoxigenic strains could be increased by the addition of exogenous cyclic AMP . A lower concentration of glucose (0.25 gm%) in culture medium produced no inhibitory effect on enzyme activity . But a higher concentration of glucose (3 gm%) repressed the tryptophanase activity . The repressive effect of glucose could be reversed by the addition of exogenous cyclic AMP. Biull Eksp Biol Med, 1985 Oct, 100(10), 472 - 4 {Inheritance and expression of the restrictive activity of plasmids coding EcoRI restrictase in Vibrio cholerae cells}; Evdokimova NM et al.; Recombinant E . coli plasmids are known to be obtained from E . coli cells using the plasmids coding EcoR1 restriction endonuclease . These plasmids were shown to possess various chromosomal or plasmid genes . The paper presents data on the construction of conjugative recombinant plasmid pSA1002, capable of conjugate transfer into V . cholerae cells . The stable maintenance and inheritance of the plasmid in V . cholerae cells have been demonstrated as well as phenotypic expression of its genes, including EcoR1 restriction endonuclease genes . The possibility of recombinant plasmids formation in V . cholerae cells dependent on EcoR1 restriction endonuclease, coded by pSA1002, is discussed. J Bacteriol, 1985 Oct, 164(1), 45 - 50 Control of Vibrio fischeri luminescence gene expression in Escherichia coli by cyclic AMP and cyclic AMP receptor protein; Dunlap PV et al.; Under certain conditions glucose represses the autoinducible synthesis of luminescence enzymes in Vibrio fischeri . To examine the genetic regulation of luminescence more closely, Escherichia coli catabolite repression mutants were transformed with a plasmid (pJE202) that contains V . fischeri genes specifying the luminescence enzymes and encoding regulatory functions for luminescence (the lux genes) or with plasmids (pJE413 and pJE455) containing transcriptional fusions between the lacZ gene on transposon mini-Mu and specific genes in each of the two lux operons . Unless cyclic AMP (cAMP) was added to the growth medium, an adenylate cyclase deletion mutant containing pJE202 produced very little light and low levels of the light-emitting enzyme luciferase . When grown in the presence or absence of cAMP, a cAMP receptor protein (CRP) deletion mutant produced low levels of light and luciferase . A mutant that does not make cAMP but does make an altered CRP which does not require cAMP for activity produced induced levels of luminescence after transformation with pJE202 . To test the effects of cAMP and CRP on each of the two lux operons separately rather than on both together, the E . coli catabolite repression mutants were transformed with pJE413 and pJE455 . From measurements of beta-galactosidase and luciferase activities it appeared that cAMP and CRP affected transcription of both lux operons . In the presence of autoinducer and its receptor, transcription of the operon encoding all of the luminescence genes except the receptor gene appeared to be activated by cAMP and CRP, whereas in the absence of the receptor, cAMP and CRP appeared to decrease transcription of this operon . Transcription of the operon encoding the autoinducer receptor appeared to be stimulated by cAMP and CRP in the absence of the receptor itself . These results demonstrate that cAMP and CRP are required for proper control of the V . fischeri luminescence system and suggest that lux gene transcription is required by a complex mechanism. Mol Gen Mikrobiol Virusol, 1985 Oct, (10), 3 - 8 {Genetic analysis of mutations affecting the synthesis of somatic O-antigen of Vibrio cholerae}; Smirnova NI et al.; The mutations oagM and oagR affecting the synthesis of somatic O-antigen have been localized on the chromosome of Vibrio cholerae El Jor and classic biotypes by conjugational crosses between different donor and recipient strains . The mutations are localized in the vicinity of the arg marker in both classic and El Tor biotypes of Vibrio cholerae. Virus Res, 1985 Oct, 3(3), 231 - 44 Isolation and characterization of influenza C virus inhibitor in rat serum; Kitame F et al.; Two hemagglutination inhibitors for influenza C virus were isolated from pooled sera of normal rats by sequential chromatography on Blue Sepharose CL 6B, Ultrogel AcA 22, and DEAE-cellulose . The two inhibitors were identified as alpha 1-macroglobulin and murinoglobulin by comparison with the authentic samples . These inhibitors abolished the hemagglutination by influenza C virus strains but did not affect the hemagglutination by influenza A and B virus strains . Hemagglutination inhibition activity of both inhibitors was completely destroyed by incubation with influenza C virus at 37 degrees C but not with the other types of influenza virus, indicating that the inhibitors are specific for influenza C virus . The inhibitory activity was also destroyed by incubation with neuraminidase from Arthrobacter ureafaciens . By contrast, no activity was lost after treatment with neuraminidase from Vibrio cholerae . These results suggest that the sialic acid residue(s) which is cleavable by the former neuraminidase but not by the latter is essential for the hemagglutination inhibition . The two inhibitors were inactivated by treating with sodium hydroxide and methylamine but not with sodium metaperiodate. Biochemistry, 1985 Sep 10, 24(19), 5235 - 40 Kinetic mechanisms of two NAD:arginine ADP-ribosyltransferases: the soluble, salt-stimulated transferase from turkey erythrocytes and choleragen, a toxin from Vibrio cholerae; Osborne JC Jr et al.; A subunit of choleragen and an erythrocyte ADP-ribosyltransferase catalyze the transfer of ADP-ribose from NAD to proteins and low molecular weight guanidino compounds such as arginine . These enzymes also catalyze the hydrolysis of NAD to nicotinamide and ADP-ribose . The kinetic mechanism for both transferases was investigated in the presence and absence of the product inhibitor nicotinamide by using agmatine as the acceptor molecule . To obtain accurate estimates of kinetic parameters, the transferase and glycohydrolase reactions were monitored simultaneously by using {adenine-2,8-3H}NAD and {carbonyl-14C}NAD as tracer compounds . Under optimal conditions for the transferase assay, NAD hydrolysis occurred at less than 5% of the Vmax for ADP-ribosylation; at subsaturating agmatine concentrations, the ratio of NAD hydrolysis to ADP-ribosylation was significantly higher . Binding of either NAD or agmatine resulted in a greater than 70% decrease in affinity for the second substrate . All data were consistent with a rapid equilibrium random sequential mechanism for both enzymes. Eur J Biochem, 1985 Sep 2, 151(2), 199 - 208 Membrane-linked energy transductions . Bioenergetic functions of sodium: H+ is not unique as a coupling ion; Skulachev VP; The concept is developed according to which Na+, like H+, can play the role of a coupling ion in energy-transducing biomembranes . This idea is based on observations that (i) Na+ can be extruded from the cell by primary pumps (Na-motive NADH-quinone reductase, decarboxylase or ATPase), and (ii) the downhill Na+ flux into the cell can be coupled with the performance of all the three types of membrane-linked work i.e . chemical (ATP synthesis), osmotic (accumulation of solutes) and mechanical (motility) . Marine alkalotolerant Vibrio alginolyticus represents the first example of such a complete sodium cycle pattern . Simplified versions of the sodium cycle or some of its constituents are found in the cytoplasmic membrane of a great variety of taxa including anaerobic, aerobic and photosynthetic bacteria, cyanobacteria and animals; this fact indicates that Na+ energetics should be regarded as a common case, rather than a rare exception applied to some natural niches only. Appl Environ Microbiol, 1985 Sep, 50(3), 724 - 6 Serotypes of Vibrio parahaemolyticus isolates from hydrobiologically dissimilar aquatic environments; Nair GB et al.; Serological analysis of the O and K antigens was performed on 324 isolates of Vibrio parahaemolyticus obtained from three hydrobiologically dissimilar aquatic environments . Only 50.9% of the strains could be serotyped . The largest number of untypable strains and the lowest serological diversity were observed from the freshwater collection . Three serotypes, O2:K28, O5:K17, and O2:K3, dominated among all biotopes . There appears to be some distinction between serotypes of environmental and clinical origins. Antimicrob Agents Chemother, 1985 Sep, 28(3), 442 - 5 In vitro susceptibility of pathogenic Vibrio species to norfloxacin and six other antimicrobial agents; Morris JG Jr et al.; The in vitro activity of norfloxacin and six other antimicrobial agents was tested against 93 vibrio strains representing the currently described pathogenic Vibrio species . Norfloxacin had excellent activity against all species, with the following MICs for 90% of the strains: 0.016 micrograms/ml for Vibrio cholerae (including tetracycline-resistant V . cholerae O1 strains), 0.25 micrograms/ml for V . parahaemolyticus, and 0.063 micrograms/ml for V . vulnificus. Zh Mikrobiol Epidemiol Immunobiol, 1985 Sep, (9), 37 - 40 {Substantiation of the necessity of developing a single set of phages for serotyping of Vibrio cholerae}; Arutiunov IuI et al.; A single set of phages for typing different V . cholerae biotypes has been developed in the USSR and proposed for use instead of two sets of phages intended for this purpose . This set has been developed in view of the following facts: the similarity of the biological properties of V . cholerae biotypes and the absence of absolute criteria for their differentiation; the existence of the foci of cholera with both biotypes circulating there; the probability experimentally confirmed, of the conversion of one biotype into the other. Vaccine, 1985 Sep, 3(3 Suppl), 211 - 4 Isolation and characterization of influenza C virus inhibitors in rat serum; Kitame F et al.; Two inhibitors against haemagglutination by influenza C virus were isolated from pooled sera of normal rats by sequential chromatography on Blue Sepharose CL 6B, Ultrogel AcA 2, and DEAE-cellulose . The two inhibitors were identified as alpha 1-macroglobulin and murinoglobulin by comparison with the authentic samples . These inhibitors abolished the haemagglutination by influenza C virus strains but did not affect the haemagglutination by influenza A and B virus strains . Haemagglutination inhibition activity of both inhibitors was completely destroyed by incubation with neuraminidase from Arthrobacter ureafaciens . By contrast, no activity was lost after treatment with neuraminidase from Vibrio cholerae . These results suggest that the sialic acid residue(s) which is excised by the former neuraminidase but not by the latter is essential for the haemagglutination inhibition . The two inhibitors were inactivated by treating with sodium hydroxide and methylamine but not with sodium metaperiodate. J Clin Microbiol, 1985 Sep, 22(3), 383 - 6 Enzyme-linked immunosorbent assays for detection of thermostable direct hemolysin of Vibrio parahaemolyticus; Honda T et al.; Several systems for enzyme-linked immunosorbent assay (ELISA) of thermostable direct hemolysin (TDH) of Vibrio parahaemolyticus were tested, and single-antibody sandwich ELISA systems gave satisfactory results . ELISA was able to detect as little as several nanograms of purified TDH per milliliter . The method of De Jong (J . Clin . Microbiol . 17:928-930, 1983) and the glutaraldehyde method were successful for preparing conjugates of alkaline phosphatase and anti-TDH antibody . TDH in fluids in intestinal loops of experimental animals challenged with living V . parahaemolyticus was accurately detectable by ELISA. J Bacteriol, 1985 Sep, 163(3), 1293 - 5 Sucrose uptake is driven by the Na+ electrochemical potential in the marine bacterium Vibrio alginolyticus; Kakinuma Y et al.; Na+ was found to be essential for the accumulation of sucrose by Vibrio alginolyticus . Sucrose uptake was completely inhibited by the addition of proton conductor at neutral pH, but not at alkaline pH, where the primary electrogenic Na+ pump generates the Na+ electrochemical gradient . We therefore conclude that sucrose transport is driven by the electrochemical potential of Na+ in this organism. J Bacteriol, 1985 Sep, 163(3), 1158 - 66 Effect of changes in the osmolarity of the growth medium on Vibrio cholerae cells; Lohia A et al.; The rate and extent of lysis of Vibrio cholerae cells under nongrowing conditions were dependent on the osmolarity of the growth medium . Gross alterations in cellular morphology were observed when V . cholerae cells were grown in media of high and low osmolarity . The rate of lysis of V . cholerae cells under nongrowing conditions increased after treatment with chloramphenicol . Chloramphenicol-treated V . cholerae 569B cells showed formation of sphaeroplast-like bodies in medium of high osmolarity, but not in low osmolarity . Changes in the osmolarity of the growth medium also regulated the expression of the outer membrane proteins . This regulation was abolished if V . cholerae cells were grown in Pi-depleted medium . Analysis of the lytic behavior and composition of outer membrane proteins of an osmotically fragile mutant strain revealed a similar dependence on the osmolarity of the growth medium. Infect Immun, 1985 Sep, 49(3), 715 - 8 Vibrio vulnificus resists phagocytosis in the absence of serum opsonins; Tamplin ML et al.; Invasive disease caused by Vibrio vulnificus may result partially from resistance to phagocytic host defense mechanisms . The present studies show that V . vulnificus resists phagocytosis by murine peritoneal macrophages in the absence of serum opsonins and extracellular bacterial products, apparently through the anti-phagocytic properties of the bacterial surface. Infect Immun, 1985 Sep, 49(3), 481 - 6 Detection of the thermostable direct hemolysin gene and related DNA sequences in Vibrio parahaemolyticus and other vibrio species by the DNA colony hybridization test; Nishibuchi M et al.; A specific gene probe for the Vibrio parahaemolyticus thermostable direct hemolysin gene was constructed and used to examine the presence or absence of the thermostable direct hemolysin gene or related DNA sequences in V . parahaemolyticus and other vibrios by the DNA colony hybridization method . The gene probe consisted of a 406-base-pair, completely internal fragment covering 71% of the structural gene with PstI linkers added to the ends . Six copies of this 415-base-pair PstI fragment were cloned into plasmid pBR322, which yielded large amounts of the probe DNA . One hundred forty-one V . parahaemolyticus strains were tested with the gene probe, and the results were compared with those of phenotypic assays for the thermostable direct hemolysin . All Kanagawa phenomenon-positive strains were gene positive . However, 86% of the strains that exhibited weak Kanagawa phenomenon and 16% of Kanagawa phenomenon-negative strains also reacted with the gene probe . Immunological methods for the detection of the thermostable direct hemolysin (modified Elek test, enzyme-linked immunosorbent assay) showed better correlation with gene probe results . All gene-positive strains produced hemolysin detectable in the enzyme-linked immunosorbent assay, although occasional strains showed weak reaction . The modified Elek test was slightly less sensitive than the enzyme-linked immunosorbent assay . All gene-negative strains were also negative in these immunological assays . One hundred twenty-one strains of Vibrio spp . other than V . parahaemolyticus were tested with the gene probe; only Vibrio hollisae strains reacted with the probe under stringent conditions. J Bacteriol, 1985 Sep, 163(3), 1210 - 4 Diffusion of autoinducer is involved in regulation of the Vibrio fischeri luminescence system; Kaplan HB et al.; The enzymes for luminescence in Vibrio fischeri are induced by the accumulation of a species-specific metabolite (autoinducer) in the culture medium . Tritium-labeled autoinducer was used to study the mechanism of autoinduction . When 3H-autoinducer was added to suspensions of V . fischeri or Escherichia coli, cellular concentrations equaled external concentrations . For V . fischeri, equilibration of 3H-autoinducer was rapid (within 20 s), and greater than 90% of the cellular tritium remained in unmodified autoinducer . When V . fischeri or E . coli cells containing 3H-autoinducer were transferred to autoinducer-free buffer, 85 to 99.5% of the radiotracer escaped from the cells, depending on the strain . Concentrations of autoinducer as low as 10 nM, which is equivalent to 1 or 2 molecules per cell, were sufficient for induction, and the maximal response to autoinducer occurred at about 200 nM . If external autoinducer concentrations were decreased to below 10 nM after induction had commenced, the induction response did not continue . Based on this study, a model for autoinduction is described wherein autoinducer association with cells is by simple diffusion and binding of autoinducer to its active site is reversible. J Clin Microbiol, 1985 Sep, 22(3), 405 - 8 New medium for the production of cholera toxin by Vibrio cholerae O1 biotype El Tor; Iwanaga M et al.; A new medium that stimulates in vitro production of cholera toxin by Vibrio cholerae O1 El Tor (El Tor vibrios) was developed . The medium contains 0.5% NaCl, 0.3% NaHCO3, 0.4% yeast extract, and 1.5% Bacto-Peptone . El Tor vibrios were cultured in a stationary test tube at 37 degrees C for 20 h . The culture supernatant was assayed for cholera toxin by a reversed passive latex agglutination method . Most vibrios grown in this medium produced 10 to 20 times more toxin than in traditional syncase medium . The number of live vibrios at the end of culture was about 10(8)/ml in the new medium (AKI medium) and about 10(10)/ml in the syncase medium . As a result, each individual organism in the new medium should have produced as much as 1,000 times more toxin than in syncase medium . Sodium bicarbonate was found to be the most important factor in toxin production by El Tor vibrios in the new medium . We recommend this new medium because of its high yield of cholera toxin and its technical simplicity. Arch Biochem Biophys, 1985 Sep, 241(2), 425 - 31 Isolation and characterization of a cyclic AMP receptor protein from luminous Vibrio harveyi cells; Chen PF et al.; A cAMP receptor protein (CRP) species was purified from the luminous Vibrio harveyi cells to apparent homogeneity . This protein had a dimeric structure with a molecular weight of 23,000 per subunit . Among all eight nucleotides tested, only cAMP (Kd = 3 to 4 microM at 0 degrees C and 52 microM at 23 degrees C) and cGMP (Kd = 6 to 10 microM at 0 degrees C and 67 microM at 23 degrees C) bound to this protein . Its binding to poly(dI-dC), poly(dA-dT), and DNA fragments isolated from V . harveyi cells were all significantly enhanced by the addition of cAMP . Based on patterns of limited proteolysis by trypsin, this CRP assumes different conformations in the absence and presence of cAMP . Also consistent with this conclusion is the finding that the binding of cAMP to CRP induced about 50% quenching of the CRP fluorescence with a concomitant 3-nm blue shift from the original 336-nm emission peak . The binding of cGMP resulted in similar fluorescence changes but had no apparent effect on the pattern of proteolysis by trypsin . Using an in vitro transcription system known to be dependent on cAMP and Escherichia coli CRP, the synthesis of a run-off transcript product was also significantly enhanced by cAMP and this V . harveyi CRP. J Bacteriol, 1985 Sep, 163(3), 1186 - 90 Functional identification of the fatty acid reductase components encoded in the luminescence operon of Vibrio fischeri; Boylan M et al.; A clone of DNA, obtained from the luminescent bacterium Vibrio fischeri ATCC 7744 and inserted into pBR322, was found to express luminescence in Escherichia coli . Polypeptides involved in biosynthesis of the fatty aldehyde substrate for the light reaction were identified by fatty acid acylation of proteins synthesized in E . coli from the recombinant plasmid . The cloned region was similar to that reported for the V . fischeri MJ1 luminescence system (Engebrecht et al., Cell 32:773-781), except for some differences in endonuclease restriction sites and the requirement of a lower temperature for the expression of light in our cloned system . Fatty acid reductase activity could be detected in extracts of E . coli harboring the recombinant plasmid but not in extracts of the parental V . fischeri strain . Using in vivo labeling with {3H}tetradecanoic acid, we showed that the acylated polypeptides synthesized in the cloned system corresponded to the labeled polypeptides in V . fischeri (34, 42, and 54 kilodaltons) and that they could only be detected after induction of luminescence . These results provide direct evidence that the genes coding for the fatty acid reductase polypeptides are an integral part of the luminescence operon in the V . fischeri luminescence system. Zh Mikrobiol Epidemiol Immunobiol, 1985 Sep, (9), 40 - 6 {Formation of drug resistance in Vibrio cholerae of non-O1 group inhabitating surface-water reservoirs}; Tichomirov ED et al.; In the study of 255 V . cholerae strains unrelated to vibrio O group I (NAG vibrios), which were isolated from water bodies in the region of the Volga delta in 1977-1982, antibioticograms of 17 types were obtained and R factor was detected in 37.7% of the strains under study . The pronounced heterogeneity of NAG vibrio populations, evaluated in minimal inhibitory concentration (MIC) values with respect to different antibiotics, is specifically manifested by the presence of various dominating subpopulations of these microorganisms, depending on their sensitivity to some chemical drugs . Various representatives of the genus Vibrio were used as a model for demonstrating the ability of the microbial population to enhance its variability as regards MIC values with a seasonal rise in the number of microorganisms. Zh Mikrobiol Epidemiol Immunobiol, 1985 Sep, (9), 26 - 9 {Allomonas--a new group of microorganisms of the Vibrionaceae family . Allomonas enterica in seawater and various hydrobiotas}; Davyborshch SG et al.; Bacteriological investigations made in the Black Sea, the Sea of Azov and the Sea of Japan have revealed that Allomonas enterica are widely spread in sea water and hydrobionts . The prevalence of these microorganisms in biotopes with unfavorable sanitary characteristics and their high correlation with the main indicator microorganisms suggest their importance as indicators, and a high percentage of seafood contaminated with Allomonas necessitates their study as possible causative agents of toxinfections. Biosci Rep, 1985 Sep, 5(9), 761 - 4 Electrophoretic resolution of microheterogeneity in Vibrio cholerae lipopolysaccharide; Ghosh S et al.; Lipopolysaccharide (LPS) from Vibrio cholerae has been analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Under normal conditions of electrophoresis which resolve Escherichia coli LPS, V . cholerae LPS shows two diffuse and unresolved bands . However, on long gels at low concentration it can be resolved into two major band types . There are at least 10 slow moving, discrete bands of regular periodicity and three fast moving bands . Comparison with LPS from E . coli indicates that the heterogeneity occurs over a much smaller range of molecular weight in V . cholerae LPS, with the entire spectrum of discrete bands being contained within the space of four E . coli repeating units. Mol Immunol, 1985 Sep, 22(9), 1123 - 9 Differences in carbohydrate specificities and complement-activating capacity of guinea pig and human antibodies to neuraminidase-treated autologous erythrocytes; Lambre CR et al.; Guinea pig erythrocytes desialated by treatment with neuraminidase from Vibrio cholerae were lyzed in autologous serum through a natural-antibody-dependent activation of the classical complement pathway . Lysis was inhibited when a mannose, glucose, galactose or N-acetyl-glucosamine was added to the incubation mixture . Methyl-alpha- or -beta-D-galactopyranosides were poorly effective and N-acetyl-D-galactosamine was not effective at all . Inhibition of lysis by the carbohydrates was due neither to an anti-complementary effect nor to a modification of the osmotic pressure since: (a) they did not alter the total complement haemolytic activity of guinea pig serum, and (b) they did not inhibit lysis of desialated guinea pig erythrocytes in human serum through activation of the alternative complement pathway . The presence of mannose, glucose, galactose or N-acetyl-glucosamine in the incubation mixture resulted in an impaired fixation of natural auto-antibodies on antigenic sites, namely the T-antigen (Thomsen-Friedenreich), which were unmasked following membrane sialic acid removal . When tested under the same conditions, only small percentage of the normal human population showed the phenomenon of lysis of desialated erythrocytes in autologous serum . Lysis was not due to a particular susceptibility of erythrocytes from these individuals to complement-mediated lysis but to the presence in their serum of complement-activating anti-T antibodies . As expected, the activity of human anti-T antibodies was inhibited by galactose and N-acetyl-galactosamine, which are the immunodominant sugars of the human T-antigen . Mannose and glucose had no effect, and methyl- alpha- or - beta-D-galactopyranosides were almost as effective as galactose . The heterogeneity of the human population with regard to the complement-activating capacity of anti-T antibodies could be of significance for the individual response of the host to an infection by a neuraminidase-producing microorganism . That the immunodominant sugars of the T-antigen were different between humans and guinea pigs was further assessed by absorption experiments . We have demonstrated that guinea pig anti-T antibodies were not removed during contact with desialated human red cells which do not have the mannose specificity, whereas human antibodies were almost entirely retained on desialated guinea pig red cells which, beside mannose, express galactose . These results also suggest that guinea pig antibodies are mostly directed towards mannose and glucose. Vet Rec, 1985 Aug 3, 117(5), 104 - 9 Post mortem studies on infertile buffalo bulls: anatomical and microbiological findings; Ahmad M et al.; Twenty-two buffalo bulls suffering from three different types of infertility were slaughtered and used for this study . Except for the reproductive system, no signs of localised or generalised disease were observed . Microbiological investigations were negative for brucellosis, vibriosis, mycoplasma and other non-specific microorganisms . Nine bulls with type 1 infertility had low bodyweights and underdevelopment of testes, accessory sex glands and endocrine glands . This picture suggests a total dysfunction of the pituitary-growth-gonadal axis . One bull of this type also showed bilateral epididymitis . Four out of 11 bulls with type 2 infertility had low bodyweights and most suffered from underdevelopment of testes, accessory sex glands and endocrine glands . Six bulls of this type had lesions of either epididymitis or orchitis or both . Two of these animals showed adhesions of periorchitis . One also showed seminal vesiculitis . In two bulls with type 3 infertility, bodyweights, reproductive organs and endocrine glands were normal . In later life, they yielded poor quality semen . Semen samples collected a few months before slaughter from nine bulls with type 2 and type 3 infertility were of poor quality and had higher percentages of abnormal spermatozoa in most cases. Appl Environ Microbiol, 1985 Aug, 50(2), 426 - 30 Incidence of Vibrio cholerae and related vibrios in a coastal lagoon and seawater influenced by lake discharges along an annual cycle; Garay E et al.; Most probable numbers of Vibrio cholerae and related vibrios were determined in Albufera Lake, Valencia, Spain, and in coastal waters under the influence of the lake discharges over the course of an annual cycle . The influence of temperature, kind of water, and characteristics of the different sampling sites on the numbers of vibrios recovered was evaluated . Maximum recovery of vibrios reached 10(3)/ml in both types of waters analyzed . V . cholerae numbers reached 10(3)/ml in the lake and 10(2) in one of the coastal sites . Frequently during the warm season, all vibrios isolated were identified as V . cholerae . Occasionally, no V . cholerae was recovered . The recovery of vibrios was significantly influenced by the temperature of the water and the type of water analyzed . Most of the V . cholerae isolates were included in Heiberg groups I and II, and nearly 50% of the strains used chitin as sole carbon source . Indole was not produced by 100% of the strains . All strains tested were non-O1 serovars. Appl Environ Microbiol, 1985 Aug, 50(2), 420 - 5 Incidence of bacteremia in stressed and unstressed populations of the blue crab, Callinectes sapidus; Welsh PC et al.; The incidence of bacteremia in the blue crab, Callinectes sapidus, is reported to be in excess of 80% . Because these results have been controversial, a field study was initiated to determine the effect of commercial capture and handling stresses on the incidence and levels of infection in blue crabs . The majority (75%) of "unstressed" crabs which were captured individually and bled immediately upon removal from the water were bacteremic, with a geometric mean level of infection of 14 CFU/ml of hemolymph . Crabs collected by crab pot, confined within these pots for as long as 24 h, and sampled immediately after removal from the water had a similar mean level of infection . Crabs subjected to the stresses of commercial capture, handling, and transport showed a higher incidence of infection (91%) and a mean infection level of 46 CFU/ml . Injuries sustained by crabs during commercial handling are thought to be associated with the higher incidence of infection . Vibrio spp . were primarily responsible for progressive infections in commercially stressed crabs and were the predominant bacterial type in heavily infected crabs . Our results indicated that uninjured healthy crabs do not have sterile hemolymph but instead harbor low-level bacterial infections. J Invest Dermatol, 1985 Aug, 85(2), 115 - 7 Appearance of dark keratinocytes following intracutaneous injection of cholera toxin in mouse skin; Murakami Y et al.; Intracutaneous injection of cholera toxin (CT), exotoxin of Vibrio cholerae, induces epidermal hyperplasia in mice, rats, and hamsters . In the work reported here we found that, like other hyperplasiogenic compounds such as 12-O-tetradecanoylphorbol-13-acetate which are tumor promoters, CT induces dark basal keratinocytes (dark cells) in the epidermis of mice . These are distinct from other epidermal cells since they contain dense cytoplasm rich in ribosomes and tonofilaments . This was demonstrated by electron microscopy and by toluidine blue staining of paraffin- or Epon-embedded sections . They comprised 3.1% of interfollicular basal cells 24-64 h after injection of 1 ng CT as compared with 0.5% in saline-injected skin . It was found by autoradiography of paraffin sections that about 47.2% of dark cells were labeled with {3H}thymidine at these times, while under the same conditions, labeling indices of basal cells were about 30% at the peaks . These results are discussed in relation to tumor promotion in two-stage carcinogenesis of mouse skin. Infect Immun, 1985 Aug, 49(2), 455 - 6 Enhancement of enterotoxin production by carbon dioxide in Vibrio cholerae; Shimamura T et al.; We found that Vibrio cholerae 569B produced much more cholera enterotoxin in the presence of added carbon dioxide than in its absence . An atmosphere of 10% carbon dioxide was optimal for maximal enterotoxin production. J Bacteriol, 1985 Aug, 163(2), 716 - 23 Flocculation in Azospirillum brasilense and Azospirillum lipoferum: exopolysaccharides and cyst formation; Sadasivan L et al.; The phenomena of flocculation and floc formation by Azospirillum brasilense Sp7 (ATCC 29145) and Azospirillum lipoferum Sp59b (ATCC 29707) were studied in aerobic liquid cultures . Carbon sources representative of various entry pathways in combination with various nitrogen sources induced flocculation in both species of azospirilla . Noticeably, the combination of fructose and nitrate was the most effective in terms of floc yields . Phase-contrast microscopic observations revealed a transition in cell morphology from freely motile, vibrioid cells to nonmotile, highly refractile encysting forms during the formation of flocs . The nonmotile forms in flocs appeared to be entangled within a fibrillar matrix, and the cells were highly resistant to desiccation . Dried flocs kept for almost 6 months still maintained the highly refractile encysting forms, and their viability was confirmed by pellicle formation and acetylene reduction in semisolid malate medium . Electron microscopic observations of the desiccated flocs revealed the presence of cell forms containing abundant poly beta-hydroxybutyrate granules within a central body and surrounded by a thick layer of exopolysaccharides . The latter were characterized by alkali and acid digestion, crude cellulase hydrolysis, and calcofluor staining . It was concluded that the overproduction of exocellular polymers induces the flocculent growth and is associated with the concomitant transformation of vegetative cells to the desiccation-resistant encysting forms under limiting cultural conditions. Can J Microbiol, 1985 Aug, 31(8), 711 - 20 Isolation and characterization of a cytolysin produced by Vibrio cholerae serogroup non-O1; McCardell BA et al.; A thermolabile toxin (molecular weight, 52 711; isoelectric point, 8.65) produced by a clinical isolate of Vibrio cholerae serogroup non-O1 was cytotoxic for Y-1 mouse adrenal cells and Chinese hamster ovary cells . The toxin lysed rabbit red blood cells and produced a hemorrhagic zone in rabbit skin . When injected intravenously into adult mice, the cytolysin was rapidly lethal and caused fluid accumulation in both 5- and 18-h rabbit ileal loops . Strains of V . cholerae that produced cytolysin but no cholerae enterotoxin were able to cause fluid accumulation in rabbit intestinal loops. J Gen Microbiol, 1985 Aug, 131 ( Pt 8), 1989 - 97 Plasmids mediating iron uptake in Vibrio anguillarum strains isolated from turbot in Spain; Tolmasky ME et al.; Vibrio strains isolated from diseased turbot in an experimental fish farm on the Atlantic coast of northwest Spain were identified as Vibrio anguillarum . The isolates shared many biochemical characteristics with V . anguillarum strains obtained from other sources, and harboured a plasmid species that showed extensive homology with plasmid pJM1, carried by V . anguillarum strain 775 isolated from an epizootic in North America . Restriction endonuclease analysis showed that the two plasmids were very similar albeit not identical . The presence of the plasmid in the turbot isolates was associated with their ability to cause disease in fish . Plasmid-carrying bacteria could also grow under conditions of iron limitation . Two outer membrane proteins, of 86 and 79 kDal, were induced, and a similar siderophore activity to that produced by V . anguillarum 775 was also detected under these conditions . The 86 kDal outer membrane protein cross-reacted immunologically with antiserum raised against the outer membrane protein OM2 produced by strain 775 . Nonvirulent plasmidless derivatives were unable to grow under iron-limiting conditions, and were also unable to produce either siderophore activity or the 86 kDal outer membrane protein, suggesting the plasmid-mediated nature of these components. J Bacteriol, 1985 Aug, 163(2), 799 - 802 Mapping of a gene that regulates hemolysin production in Vibrio cholerae; von Mechow S et al.; A gene that regulates the hemolysin structural gene (hly) was found to be tightly linked to the tox-1000 locus of Vibrio cholerae RJ1 and separated from hly by a large section of the V . cholerae genetic map . This hemolysin regulatory gene was designated hlyR. J Bacteriol, 1985 Aug, 163(2), 580 - 5 Genetic analysis of the cholera toxin-positive regulatory gene toxR; Miller VL et al.; Southern blot analysis with a toxR-specific gene probe indicates that Vibrio cholerae 569B has a 1.2-kilobase deletion near the toxR gene . Heterologous conjugative crosses were carried out between the EI Tor strain RV79 and 569B tox mutants . Tox+ recombinants showed the same linkage properties to the his locus as to the previously mapped tox locus of 569B . Southern blot analysis with the toxR probe of the Tox+ recombinants obtained in these heterologous crosses showed that these recombinants had replaced the V . cholerae 569B (recipient) toxR DNA with the V . cholerae RV79 (donor) toxR DNA, indicating that tox and toxR are the same locus . However, the Tox+ recombinants synthesized an amount of toxin intermediate between the level observed for wild-type RV79 and 569B strains, suggesting there is a difference in the ability of toxR genes from different strains to activate ctx . About half of the mutations which suppress the phenotype of hypertoxinogenic locus htx are unlinked to htx and in addition have a hypotoxinogenic phenotype relative to that of the wild type . Most of these hypotoxinogenic, second-site suppressors show a linkage to his similar to the linkage of toxR to his and are therefore probably mutations in toxR . These results indicate that the toxR gene product is required for ctx expression and that a functional toxR gene is required for the effect of an htx mutation to be seen. Infect Immun, 1985 Aug, 49(2), 275 - 80 Immunological characterization of Vibrio cholerae O:1 lipopolysaccharide, O-side chain, and core with monoclonal antibodies; Gustafsson B et al.; Lipopolysaccharide (LPS) was extracted from Vibrio cholerae O:1 strains of the serotypes Ogawa, Inaba, and Hikojima and delipidated by mild-acid hydrolysis . Two polysaccharide fragments with the molecular weights of approximately 9,000 and 900, respectively, were isolated by gel permeation chromatography . The LPS preparations and the polysaccharide fragments were studied in enzyme-linked immunosorbent assay inhibition, rocket immunoelectrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroblotting with monoclonal antibodies directed against the group-specific antigen A, the type-specific antigens B (Ogawa) and C (Inaba), and the core region . Antigen A was demonstrated in all LPS preparations and all 9,000-molecular-weight fragments tested . The type-specific antigens B and C were demonstrated in LPSs and 9,000-molecular-weight fragments from Ogawa and Inaba, respectively . Furthermore, antigens B and C were both demonstrated in LPSs and 9,000-molecular-weight fragments from two of four Hikojima strains tested . Core antigen was demonstrated in the LPS and in the 9,000- and 900-molecular-weight fragments . The results indicate that the 9,000-molecular-weight fragment represents the complete polysaccharide chain, including group- and type-specific antigens as well as core antigens, whereas the 900-molecular-weight fragment constitutes the main part of the core region. J Mol Biol, 1985 Jul 5, 184(1), 179 - 81 Preliminary crystallographic study of an L-asparaginase from Vibrio succinogenes; Ammon HL et al.; Crystals of an L-asparaginase from Vibrio succinogenes were obtained with the hanging drop method from ammonium sulphate-containing solutions . The crystals belong to the orthorhombic space group P22(1)2(1) with unit cell dimensions of a = 71.3 A, b = 85.8 A, c = 114.0 A, and contain two tetrameric enzyme molecules per unit cell . There are two subunits in the asymmetric unit; a molecular dyad is coincident with the crystallographic dyad . The crystal lattice is similar to that reported for an Escherichia coli asparaginase . Rotation function calculations have revealed that the V . succinogenes enzyme has 222 point group symmetry in the crystal . The second and third molecular dyads differ, however, from the corresponding E . coli asparaginase dyads by approximately 40 degrees . The crystals diffract to at least 2.2 A resolution and are suitable for X-ray crystallographic structure determination. Am J Dis Child, 1985 Jul, 139(7), 701 - 4 Peritoneal neutrophilic cell response in necrotizing enterocolitis; Balcom RJ et al.; Neutropenia commonly occurs in neonates with necrotizing enterocolitis (NEC) . In an attempt to determine the etiology of this neutropenia, we observed the peripheral and peritoneal neutrophil cell responses of seven infants at the time of surgery . Six of seven patients had diminished peripheral neutrophil counts within 24 hours prior to surgery, with substantial mobilization of mature neutrophils into the peritoneum . We also looked at the rat peritoneum as a model for neutrophilic cell consumption using casein and Vibrio cholerae enterotoxin to cause neutrophil mobilization . With both agents, significant mobilization of neutrophils into the peritoneum occurred . Bone marrow stores in the animals were substantially decreased, but neutropenia was not observed . We speculate that the neutropenia of NEC is largely a consequence of neutrophil mobilization into the peritoneum, perhaps initiated by dietary protein and/or bacterial toxin. Genetika, 1985 Jul, 21(7), 1090 - 8 {Construction of a genetic map of the chromosome of Vibrio cholerae based on conjugational crossings}; Smirnova NI et al.; The results of cholera vibrio chromosomal mapping using Vibrio cholerae classica and V . cholerae eltor donor strains obtained with the help of various R . plasmids, are summarized in the paper . A genetic map of V . cholerae chromosome was established showing the order of 35 gene markers . The relationship between the genetic structures of cholera eltor and classical vibrio biotypes is discussed. Cancer Res, 1985 Jul, 45(7), 3048 - 52 Effects of pyrimidine antagonists on sialic acid regeneration in HL-60 cells; Hindenburg AA et al.; Because alterations in cell membrane sialoglycoconjugates can affect the behavior of neoplastic cells, we investigated the effects of in vitro treatment with antimetabolites used in cancer therapy on the expression of membrane sialic acid in cultured HL-60 leukemic cells . In these studies, cells were incubated with Vibrio cholerae neuraminidase to remove surface sialic acid . Reappearance of membrane sialic acid during drug treatment was followed (a) by measuring changes in radioactive surface labeling of viable cells with sodium metaperiodate-sodium{3H}-borohydride, (b) by measuring the decline in accessible surface galactosyl receptor sites which occurred coincident with membrane sialic acid replacement, and (c) by measuring the incorporation of {3H}glucosamine into membrane-associated neuraminidase-labile sialic acid . We were especially interested in learning whether drugs that affect intracellular pools of cytidine triphosphate (CTP), an important nucleotide intermediate in sialylation reactions, could inhibit regeneration of membrane sialic acid . 3-Deazauridine, a competitive inhibitor of CTP synthetase, depleted CTP pools and curtailed surface membrane resialylation with little or no effect on synthesis of de novo sialic acid from precursor sugars . The addition of cytidine restored CTP pools and sialic acid regeneration . Acivicin, a glutamine antagonist, also depleted CTP pools and curtailed surface membrane resialylation . In addition, it retarded de novo synthesis of sialic acid . The addition of cytidine restored intracellular CTP pools and sialic acid regeneration . However, both cytidine and guanosine were required to restore sialic acid synthesis from precursor sugars . 1-beta-D-Arabinofuranosylcytosine, a competitive inhibitor of sialic acid synthetase and of sialyltransferase, inhibited both de novo sialic acid synthesis and membrane resialylation . Only the latter effect was reversed by the addition of exogenous cytidine . Hydroxyurea, an agent shown previously to inhibit glycoconjugate production in hamster fibroblasts, curtailed membrane resialylation and de novo synthesis of sialic acid without depleting CTP pools . Doxorubicin, at levels that caused marked arrest of cell proliferation, had no effect on sialic acid synthesis or expression on the membrane surface . These data suggest that antimetabolites, apart from their cytotoxic effects or effects on cellular growth, may directly inhibit the expression of membrane sialic acid.(ABSTRACT TRUNCATED AT 400 WORDS) Infect Immun, 1985 Jul, 49(1), 122 - 31 Monoclonal antibodies to outer membrane antigens of Vibrio cholerae; Sciortino CV et al.; Hybridoma-derived monoclonal antibodies were prepared against outer membrane antigens of four strains of Vibrio cholerae that were cultivated under iron-limited conditions, and these antibodies were partially characterized . We established a library of 66 hybridomas which produced monoclonal antibodies defining 16 different V . cholerae antigens . Two antigens (molecular weights, 18,000 and 112,000) were heat modifiable, whereas the reacting epitope of a third antigen (40,000-dalton-18,000-dalton doublet) was completely destroyed when it was heated at 100 degrees C . The 112,000-dalton heat-modifiable protein was an iron-regulated outer membrane protein . This protein bound 59Fe in vitro when it was combined with the V . cholerae siderophore-iron complex 59Fe-vibriobactin; it was also found in in vivo grown V . cholerae, as were three other antigens . A total of 26 hybridomas produced antibody to V . cholerae lipopolysaccharide . Of these, 12 were cross-reactive with lipopolysaccharides of other gram-negative bacteria, including 2 which recognized lipid A . Several of these anti-lipopolysaccharide monoclonal antibodies appeared to be lipopolysaccharide region specific . Some membrane antigens were strain specific, whereas others were common to both O group 1 and non-O group 1 vibrios. J Wildl Dis, 1985 Jul, 21(3), 211 - 8 Detection of Vibrio anguillarum antigen by the dot blot assay; Cipriano RC et al.; The dot blot assay, modified and adapted for detection of antigens from Vibrio anguillarum in fish tissues, was specific for V . anguillarum and did not react with antigens of V . ordalii, Pseudomonas sp., or Yersinia ruckeri . The blot assay enabled detection of as little as 2.3 ng of a mixture of protein antigens obtained from cell-free extracts of V . anguillarum; it was about 100 times more sensitive than either the indirect fluorescent antibody technique or bacterial isolation for detecting V . anguillarum in fish tissues. J Biol Chem, 1985 Jun 10, 260(11), 6938 - 44 Purification and characterization of a bioluminescence-related fatty acyl esterase from Vibrio harveyi; Byers D et al.; Vibrio harveyi extracts contain three polypeptides (32, 42, and 57 kDa) which are involved in long-chain aldehyde biosynthesis and can be labeled with {3H} tetradecanoic acid (+ATP) and/or {3H}tetradecanoyl-CoA . These proteins have been separated from other labeled bands by ammonium sulfate fractionation, and the 32-kDa polypeptide has been further purified to homogeneity by ion-exchange, gel filtration, and hydroxylapatite chromatography . In aqueous buffers at pH 7, the 32-kDa protein catalyzes the hydrolysis of tetradecanoyl-CoA at a low rate (0.01 mumol/min/mg) to form free fatty acids . The thioesterase rate is slightly increased by phosphate, which also protects the enzyme against inhibition by the sulfhydryl reagent N-ethylmaleimide . Acyl-CoA cleavage is dramatically stimulated (up to 100-fold) by certain organic solvents, in particular glycerol and ethylene glycol, with the fatty acyl group being transferred to the alcohol acceptors . These enzymatic properties may be related to the role of the 32-kDa esterase in generating fatty acids for subsequent use in the V . harveyi bioluminescent system. Southeast Asian J Trop Med Public Health, 1985 Jun, 16(2), 265 - 7 An evaluation of alkaline peptone water for enrichment of Vibrio cholerae in feces; Lesmana M et al.; A comparison was made to determine the sensitivity of direct inoculation of thiosulfate citrate bile salts agar (TCBS) and alkaline peptone water (APW) enrichment prior to direct inoculation of TCBS to culture Vibrio cholerae from feces of patients with gastroenteritis . V . cholerae was isolated from 611 feces specimens . Of those, 535 were isolated in TCBS and APW-TCBS, 15 in only TCBS and 61 in only APW-TCBS . V . parahemolyticus (21) and non-agglutinating vibrios (11) were also isolated but more often in direct inoculated TCBS than APW-TCBS cultures . Maximum isolation sensitivity of V . cholerae and V . parahemolyticus from feces is obtained by both direct inoculation of TCBS and enrichment in APW prior to TCBS inoculation. J Clin Microbiol, 1985 Jun, 21(6), 884 - 90 In vitro and in vivo cholera toxin production by classical and El Tor isolates of Vibrio cholerae; Turnbull PC et al.; A comparative study was carried out on the in vitro production of cholera toxin by 19 Vibrio cholerae El Tor isolates from patients with cholera in South Africa, one El Tor isolate from a patient in Malawi (a country approximately 1000 km north-northeast of South Africa), 6 El Tor and 12 classical type isolates from patients in Bangladesh, and 5 culture collection classical strains . Identical phage types and indistinguishable toxigenicities among the South African and Malawi V . cholerae, representing isolations obtained over a 10-year period, indicated that essentially a single strain was involved in the cholera of these regions . Similarly, phage typing and toxin profiles indicated that the 12 classical and 6 El Tor V . cholerae cultures in Bangladesh, all isolated in November 1983, represented just two strains . As assessed by titrations in Y-1 mouse adrenal and Chinese hamster ovary cell lines, the general order of toxigenicities was Bangladesh and culture collection classical greater than Bangladesh El Tor greater than southern African El Tor . The African isolates consistently gave rise to very low titers . Their relative reluctance to produce the toxin in vitro compared with the culture collection classical strains, particularly strain 569B, was confirmed by rocket electrophoresis . In somewhat of a contrast, maximum in vivo titers in rice water stools from cholera patients in South Africa and from both classical and El Tor type cholera patients in Bangladesh were essentially equal . It is postulated that under the continuous culture conditions that occur in vivo, cholera toxin concentrations can accumulate to a maximum level, depending on the rate of purging by the diarrheal fluid rather than the toxigenicity of the infecting stain . The relevance of these findings to the relative severities of classical and El Tor types of cholera is discussed. Southeast Asian J Trop Med Public Health, 1985 Jun, 16(2), 261 - 4 A CAMP phenomenon between Vibrio cholerae biotype El Tor and staphylococcal B-hemolysin; Lesmana M et al.; A CAMP phenomenon was demonstrated by Vibrio cholerae biotype El Tor and B-lysin producing Staphylococcus aureus in 5% sheep red blood cells-tryptic soy agar medium . All 394 El Tor vibrio strains tested, all showed a crescent-shaped hemolysis (positive CAMP) when the cultures were incubated in a candle jar whereas 67% were CAMP positive when incubated aerobically . Only 9% of the isolates produced detectable hemolysin in a standard tube test using heart infusion broth and 72% in a tube test using heart infusion broth containing 1% glycerol . Seven classical V . cholerae tested were CAMP negative . The CAMP reaction is easy to perform and may be useful for routine use in the differentiation of V . cholerae biotype El Tor from classical V . cholerae. Infect Immun, 1985 Jun, 48(3), 754 - 8 Evaluation of BW942C, a novel antidiarrheal agent, against enterotoxins of Escherichia coli and Vibrio cholerae; Morgan DR et al.; BW942C, an enkephalin-like pentapeptide with anti-diarrheal activity, was tested against crude toxins of Escherichia coli and Vibrio cholerae in the Y-1 adrenal cell assay, rabbit ileal loop assay, and suckling mouse assay . The effects of BW942C on in vitro ion transport were measured in rabbit ileum mounted in Ussing chambers . In vitro, BW942C decreased basal short-circuit current (2.26 and 3.15 mueq cm-2 h-1 in experimental samples and controls, respectively; n = 7, P less than 0.05) and increased basal net Cl absorption (1.59 and 0.50 mueq cm-2 h-1 in experimental samples and controls, respectively; P less than 0.025) . Net Na absorption was also increased, but not significantly . BW942C did not block the secretory response to a maximal dose of purified heat-stable toxin . BW942C directly enhanced intestinal fluid absorption . In the Y-1 adrenal cell assay, 5 mg of BW942C per ml inhibited the cytopathic effect caused by cholera toxin or heat-labile enterotoxin of E . coli . In the rabbit ileal loop assay, E . coli heat-stable toxin, E . coli heat-labile enterotoxin, and cholera toxin were inhibited 35 to 70% by administration of BW942C . With the suckling mouse model, the fluid accumulation caused by E . coli heat-stable toxin was ablated by prior treatment with BW942C . The drug is currently being evaluated in patients with acute secretory diarrhea to determine its effect on clinical symptoms. J Biol Chem, 1985 May 25, 260(10), 6139 - 46 Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the alpha subunit of bacterial luciferase; Cohn DH et al.; The nucleotide sequence of the 1.85-kilobase EcoRI fragment from Vibrio harveyi that was cloned using a mixed-sequence synthetic oligonucleotide probe (Cohn, D . H., Ogden, R . C., Abelson, J . N., Baldwin, T . O., Nealson, K . H., Simon, M . I., and Mileham, A . J . (1983) Proc . Natl . Acad . Sci . U.S.A . 80, 120-123) has been determined . The alpha subunit-coding region (luxA) was found to begin at base number 707 and end at base number 1771 . The alpha subunit has a calculated molecular weight of 40,108 and comprises a total of 355 amino acid residues . There are 34 base pairs separating the start of the alpha subunit structural gene and a 669-base open reading frame extending from the proximal EcoRI site . At the 3' end of the luxA coding region there are 26 bases between the end of the structural gene and the start of the luxB structural gene . Approximately two-thirds of the alpha subunit was sequenced by protein chemical techniques . The amino acid sequence implied by the DNA sequence, with few exceptions, confirmed the chemically determined sequence . Regions of the alpha subunit thought to comprise the active center were found to reside in two discrete and relatively basic regions, one from around residues 100-115 and the second from around residues 280-295. JAMA, 1985 May 17, 253(19), 2850 - 3 Vibrio vulnificus . Man and the sea; Johnston JM et al.; To identify risk factors for Vibrio vulnificus infections, we performed a regional case-control study of 19 patients identified by isolates received at a state reference laboratory . Interviews with patients or surviving relatives and with three controls for each patient were compared in a matched analysis . Patients with V vulnificus wound infection were more likely than controls to have sustained a puncture wound while handling fresh seafood or to have been exposed to salt water . More patients with primary septicemia than controls had eaten raw oysters before the onset of illness . Other risk factors for septicemia included underlying liver disease, hematopoietic disorders, chronic renal insufficiency, use of immunosuppressive agents, and heavy alcohol consumption . Although V vulnificus infection is unusual, with a regional incidence of 0.8 per 100,000 population in this study, septicemia in the immunosuppressed patient is a devastating illness that can be prevented by not eating raw seafood. Acta Pathol Jpn, 1985 May, 35(3), 731 - 9 Vibrio vulnificus septicemia; Shirouzu K et al.; A 33-year-old Japanese male, who had a three year history of biopsy-proved liver cirrhosis, was admitted to the hospital on June, 24, 1983 with a sudden onset of fever (38.6 degrees C), chills, generalized pain, nausea, anorexia, weakness, and eruption over the entire body . The patient went into shock and died about 7 hours after admission . Blood cultures before death were positive for V . vulnificus . Postmortem microscopic examination revealed "necrotizing vasculitis" in the small and large intestines, stomach, and skin, and also showed marked toxic epidermal necrolysis . This case matches the primary septicemia caused by V . vulnificus described by Blake et al . In addition, this case suggests that the septicemia was acquired through the gastrointestinal tract, especially the small intestine, because the V . vulnificus was isolated from blood and numerous Gram-negative bacilli around the submucosal vessels were observed in the area with acute necrotizing vasculitis. Appl Environ Microbiol, 1985 May, 49(5), 1232 - 4 Properties of lactate dehydrogenase in a psychrophilic marine bacterium; Mitchell P et al.; Lactate dehydrogenase (EC 1.1.1.27) from Vibrio marinus MP-1 was purified 15-fold and ammonium activated . The optimum pH for pyruvate reduction was 7.4 . Maximum lactate dehydrogenase activity occurred at 10 to 15 degrees C, and none occurred at 40 degrees C . The crude-extract enzyme was stable between 15 and 20 degrees C and lost 50% of its activity after 60 min at 45 degrees C . The partially purified enzyme was stable between 8 and 15 degrees C and lost 50% of its activity after 60 min at 30 degrees C . The thermal stability of lactate dehydrogenase was increased by mercaptoethanol, with 50% remaining activity at 42 degrees C. J Neurochem, 1985 May, 44(5), 1378 - 84 A new rapid and sensitive bioluminescence assay for monoamine oxidase activity; Tenne M et al.; The in vivo luminescence of an aldehyde-requiring mutant of the luminous bacteria Vibrio harveyi (M42) increases dramatically upon the addition of long-chain aliphatic aldehydes (C8-C16) . The intensity of this luminescence is linearly related to aldehyde concentration . This property was utilized for the determination of monoamine oxidase activity using n-octylamine and n-decylamine as substrates, which are converted by monoamine oxidase to n-octylaldehyde and n-decylaldehyde, respectively . The addition of the amine to a suspension containing rat liver mitochondria and M42 cells initiated a luminescence that was directly proportional to monoamine oxidase activity according to two parameters: (1) the rate of the initial increase in luminescence and (2) the final "steady-state" level of luminescence . The new assay has advantages of high sensitivity, rapidity, the possibility to perform discontinuous as well as continuous monitoring of monoamine oxidase activity, and applicability to turbid preparations. J Bacteriol, 1985 May, 162(2), 558 - 64 Nucleotide sequence of the thermostable direct hemolysin gene of Vibrio parahaemolyticus; Nishibuchi M et al.; The gene encoding the thermostable direct hemolysin of Vibrio parahaemolyticus was characterized . This gene (designated tdh) was subcloned into pBR322 in Escherichia coli, and the functional tdh gene was localized to a 1.3-kilobase HindIII fragment . This fragment was sequenced, and the structural gene was found to encode a mature protein of 165 amino acid residues . The mature protein sequence was preceded by a putative signal peptide sequence of 24 amino acids . A putative tdh promoter, determined by its similarity to concensus sequences, was not functional in E . coli . However, a promoter that was functional in E . coli was shown to exist further upstream by use of a promoter probe plasmid . A 5.7-kilobase SalI fragment containing the structural gene and both potential promoters was cloned into a broad-host-range plasmid and mobilized into a Kanagawa phenomenon-negative V . parahaemolyticus strain . In contrast to E . coli, where the hemolysin was detected only in cell lysates, introduction of the cloned gene into V . parahaemolyticus resulted in the production of extracellular hemolysin. Blood, 1985 May, 65(5), 1120 - 6 Sialic acid in mature megakaryocytes: detection by wheat germ agglutinin; Schick PK et al.; The characteristics of the surface of guinea pig megakaryocytes were investigated with wheat germ agglutinin (WGA) . Purified guinea pig megakaryocytes and platelets were incubated with WGA conjugated with rhodamine, cytocentrifuged, and then exposed to Chromomycin A3 for the assessment of ploidy . The fluorescence emission of the DNA-Chromomycin complex was similar to that of fluorescein, and thus both rhodamine-WGA and Chromomycin A3 fluorescence could be analyzed in the same cell . Ploidy was assessed by microdensitometry of Chromomycin A3 fluorescence . Eight hundred megakaryocytes were analyzed by four parameters: (1) labeling by WGA, (2) ploidy, (3) morphological stage, and (4) size . The results were analyzed by a computer-assisted program . Although all platelets had reacted with WGA, only about half of the isolated megakaryocytes had been labeled by the lectin . The analysis of the megakaryocytes that had reacted revealed that 72% of stage III and 77% of stage IV megakaryocytes as compared with 35% of stage I and 29% of stage II cells had been labeled by the lectin . WGA reacted with 44% of 8N megakaryocytes and 60% and 59% of 16N and 32N cells . However, WGA labeling was independent of megakaryocyte size . The digestion of 15% and 48% of megakaryocyte sialic acid with neuraminidase from Vibrio cholera resulted in a 67% and 89% decrease in the binding of rhodamine-WGA, respectively, as determined by microdensitometry . The study indicated that sialic acid serves as a receptor for WGA and that sialoglycoproteins and possibly gangliosides become exposed on the surface of mature megakaryocytes . WGA can recognize mature megakaryocytes by biochemical criteria and the assessment of lectin binding could complement the morphological staging of megakaryocytes. Diagn Microbiol Infect Dis, 1985 May, 3(3), 223 - 32 Biochemical and exoenzymatic properties of Aeromonas species; Janda JM; One hundred twenty-seven isolates of Aeromonas comprising the three currently recognizable species (A . hydrophila, A . sobria, and A . caviae) were evaluated for biochemical and exoenzymatic properties . Aeromonas species were generally (greater than 90%) characterized as gram-negative fermentative rods that were oxidase-, catalase-, and beta-galactosidase-positive, produced arginine dihydrolase, and failed to decarboxylate ornithine . More than 95% of all isolates tested failed to grow on 6.5% salt or thiosulfate-citrate bile salts agar and were resistant to the vibriostatic agent 0/129 . Most Aeromonas species produced acid from hexoses while failing to ferment alcoholic sugars or trisaccharides . In exoenzymatic studies, Aeromonas species were uniformly found to produce several exoenzymes, including amylase, DNase, RNase, esterase, lipase, gelatinase, protease, fibrinolysin, and chitinase . Within the genus, a number of biochemical and enzymatic properties were found to be associated with one or more of the taxonomically recognizable species . These properties included glycoside utilization, Heiberg grouping based upon fermentation of arabinose, sucrose, and mannose, and the elaboration of several extracellular enzymes (elastase, hemolysin, lecithinase, phosphatase) . In addition, phenotypic markers previously associated with enterotoxigenic Aeromonas isolates were almost exclusively found among A . hydrophila and A . sobria species, suggesting that these species are the major enteric pathogens. J Bacteriol, 1985 May, 162(2), 510 - 5 Cloning and expression in Escherichia coli of Vibrio parahaemolyticus thermostable direct hemolysin and thermolabile hemolysin genes; Taniguchi H et al.; Two hemolysin genes of Vibrio parahaemolyticus WP1, a thermostable direct (TSD) hemolysin gene and a thermolabile hemolysin gene, were cloned into the pBR322 vector in Escherichia coli K-12 C600 . A large amount of the TSD hemolysin produced in E . coli K-12 accumulated in the periplasmic space . The TSD hemolysin gene was localized on a 0.9-kilobase HindIII-BamHI fragment by identifying qualitatively the production of the TSD hemolysin by a reverse passive hemagglutination assay in the osmotic shock fluid . The thermolabile hemolysin gene was isolated on a 1.3-kilobase HindIII-PstI fragment by selection with the hemolysin on blood agar . Southern blot hybridization and colony hybridization experiments indicated that the TSD hemolysin gene was present in the chromosomal DNA of 15 Kanagawa phenomenon-positive strains but not in 14 negative strains, whereas the thermolabile hemolysin gene was detected in all strains . No homologous DNA sequences to TSD and thermolabile hemolysin genes were detected in the chromosomes of Vibrio cholerae, Vibrio vulnificus, non-O1 V . cholerae, and Vibrio anguillarum. J Bacteriol, 1985 May, 162(2), 794 - 8 Respiration-driven Na+ pump and Na+ circulation in Vibrio parahaemolyticus; Tsuchiya T et al.; Sodium circulation in Vibrio parahaemolyticus was investigated . We observed respiration-driven Na+ extrusion from cells by using a Na+ electrode . The Na+ extrusion was insensitive to a proton conductor, carbonyl cyanide m-chlorophenylhydrazone, and sensitive to a respiratory inhibitor, CN- . These results support the idea of the existence of a respiratory Na+ pump in V . parahaemolyticus . The respiration-driven Na+ extrusion was observed only under alkaline conditions. Eur J Biochem, 1985 Apr 15, 148(2), 385 - 90 Purification of the 25-kDa Vibrio cholerae major outer-membrane protein and the molecular cloning of its gene: ompV; Stevenson G et al.; The 25-kDa peptidoglycan-associated outer-membrane protein and most likely porin of Vibrio cholerae is a major immunogenic species . It has been purified by ion-exchange elution on hydroxyapatite followed by gel filtration on Bio-Gel P150 both in the presence of sodium dodecyl sulfate . This protein, of greater than 90% purity as judged by Western blotting, has been used to raise antibodies in rabbits . The antisera were then used to screen V . cholerae gene banks, constructed in Escherichia coli K12, and this has enabled us to isolate several colonies harbouring the cloned gene . The plasmids in these colonies have been designated pPM451, pPM455 and pPM472 . These plasmids have a 5.3 X 10(3)-base BamHI fragment of V . cholerae DNA in common . Restriction endonuclease mapping of these plasmids has been performed and the protein identified both by Western blot analysis and in E . coli K12 minicells . The protein is not efficiently expressed in E . coli K12 . It is proposed to use the name ompV to describe the structural gene, present in the cloned DNA, for this V . cholerae outer membrane protein. FEBS Lett, 1985 Apr 8, 183(1), 95 - 8 Generation of the electrochemical potential of Na+ by the Na+-motive NADH oxidase in inverted membrane vesicles of Vibrio alginolyticus; Tokuda H et al.; Inverted membrane vesicles prepared from Vibrio alginolyticus generated a membrane potential (positive inside) and accumulated Na+ by the oxidation of NADH . Generation of the membrane potential required Na+ and was inhibited by 2-heptyl-4-hydroxyquinoline N-oxide, a specific inhibitor of the Na+-dependent NADH oxidase . Collapse of the membrane potential by valinomycin stimulated the uptake of Na+ . In contrast, accumulation of H+ was not detected under all the conditions tested . These results suggest that only Na+ is translocated by the Na+-dependent NADH oxidase of V . alginolyticus. Xenobiotica, 1985 Apr, 15(4), 271 - 6 The inhibition of bacterial bioluminescence by xenobiotics; Danilov VS et al.; The effect of various xenobiotic substrates of microsomal cytochrome P-450, including dimethylaniline, ethylmorphine, hexabarbital and aminopyrine, on the bioluminescence of the bacteria Vibrio fischeri and the bacterial luciferase mixed-function oxidase system is described . These compounds are effective inhibitors of the luminescence reaction . The inhibition provided evidence for the competitive nature of the interactions between xenobiotics and an aliphatic aldehyde, which is a substrate of bacterial luciferase, at the binding site for cytochrome P-450 . The bioluminescence method is suitable for the analysis of metabolism and detoxication of various xenobiotics. J Appl Bacteriol, 1985 Apr, 58(4), 407 - 23 A probability matrix for the identification of vibrios; Dawson CA et al.; A probability matrix for computer-assisted identification of vibrios has been constructed, based on the API 20E system . Data were gathered from 173 strains representing 31 taxa of vibrios and related organisms, from a variety of sources . The matrix was tested internally by four statistical programs . Program OVERMAT tested the separation and program MOSTTYP the discretion and homogeneity of the taxa . Most of the taxa were satisfactory but a few were less so; reasons for this are discussed . Program CHARSEP and program DIACHAR tested the separation and diagnostic values, respectively, of the characters used . The overall test error was 4.5% . The matrix was assessed externally by its performance in the identification of vibrio-like strains isolated from freshwater . Of 243 wild strains, 79.4% were identified with ten taxa, with a Willcox score of greater than or equal to 0.99. Infect Immun, 1985 Apr, 48(1), 87 - 93 Identification and characterization of Vibrio cholerae surface proteins by radioiodination; Richardson K et al.; Whole cells and isolated outer membrane from Vibrio cholerae (Classical, Inaba) were radiolabeled with Iodogen or Iodo-beads as catalyst . Radiolabeling of whole cells was shown to be surface specific by sodium dodecyl sulfate-urea polyacrylamide gel electrophoresis of whole cells and cell fractions . Surface-labeled whole cells regularly showed 16 distinguishable protein species, of which nine were found in radiolabeled outer membrane preparations obtained by a lithium chloride-lithium acetate procedure . Eight of these proteins were found in outer membranes prepared by sucrose density gradient centrifugation and Triton X-100 extraction of radiolabeled whole cells . The mobility of several proteins was shown to be affected by temperature, and the major protein species exposed on the cell surface was shown to consist of at least two different peptides. J Bacteriol, 1985 Apr, 162(1), 209 - 16 Mechanisms of iron regulation of luminescence in Vibrio fischeri; Haygood MG et al.; Synthesis of luciferase (an autoinducible enzyme) is repressed by iron in the symbiotic bioluminescent bacterium Vibrio fischeri . Possible mechanisms of iron regulation of luciferase synthesis were tested with V . fischeri and with Escherichia coli clones containing plasmids carrying V . fischeri luminescence genes . Experiments were conducted in complete medium with and without the synthetic iron chelator ethylenediamine-di(o-hydroxyphenyl acetic acid) . Comparison of the effect of ethylenediamine-di(o-hydroxyphenyl acetic acid) and another growth inhibitor, (2-n-heptyl-4-hydroxyquinoline-N-oxide), showed that iron repression is not due to inhibition of growth . A quantitative bioassay for autoinducer was developed with E . coli HB101 containing pJE411, a plasmid carrying V . fischeri luminescence genes with a transcriptional fusion between luxI and E . coli lacZ . Bioassay experiments showed no effect of iron on either autoinducer activity or production (before induction) or transcription of the lux operon . Ethylenediamine-di(o-hydroxyphenyl acetic acid) did not affect luciferase induction in E . coli strains with wild-type iron assimilation (ED8654) or impaired iron assimilation (RW193) bearing pJE202 (a plasmid with functional V . fischeri lux genes), suggesting that the genes responsible for the iron effect are missing or substituted in these clones . Two models are consistent with the data: (i) iron represses autoinducer transport, and (ii) iron acts through an autoinduction-independent regulatory system (e.g., an iron repressor). J Bacteriol, 1985 Apr, 162(1), 35 - 41 Cloning and characterization of the hemolysin determinants from Vibrio cholerae RV79(Hly+), RV79(Hly-), and 569B; Goldberg SL et al.; The Hly region from the chromosome of Vibrio cholerae El Tor strain RV79(Hly-) and the nonhemolytic classical strain 569B were cloned into plasmid vector pBR322 . Escherichia coli K-12 transformants possessing these recombinant plasmids were nonhemolytic and were detected with a 32P-labeled hly-specific DNA probe . Restriction endonuclease Sau3AI digestions of the cloned hly loci of two independently obtained RV79(Hly+) convertants, when compared with the digests of cloned RV79(Hly-) loci, revealed that an apparent alteration (10 to 15 base pairs) had occurred . In contrast, an apparent 20-base-pair deletion was present in the cloned hly locus of the classical biotype V . cholerae strain 569B . Maxicell analysis and immunoprecipitation of labeled proteins of E . coli which are encoded by the cloned hly loci of RV79(Hly+) and from nuclease BAL 31-deleted plasmids, as well as immunoprecipitation of {35S}methionine-labeled V . cholerae proteins, suggest that the hemolysin is an 84,000-dalton polypeptide. Biochem Biophys Res Commun, 1985 Mar 29, 127(3), 1007 - 11 Expression of the cloned subunits of bacterial luciferase from separate replicons; Gupta SC et al.; The lux A and lux B genes of Vibrio harveyi, encoding the alpha and beta subunits of bacterial luciferase, were cloned individually into Escherichia coli in two different compatible plasmids . Active luciferase was formed in an amount equal to that produced in cells carrying a plasmid with the cloned genes on a single fragment. Science, 1985 Mar 15, 227(4692), 1345 - 7 Measuring gene expression with light; Engebrecht J et al.; Light is produced by recombinant Escherichia coli that contain lux genes cloned from the marine bacterium Vibrio fischeri . The bioluminescence phenotype requires genes for regulatory and biochemical functions, the latter encoded by five lux genes contained in a single operon . These lux genes were disconnected from their native promoter and inserted into the transposon mini-Mu . The resulting transposon, mini-Mulux, could induce mutations by insertional inactivation of a target gene, and the lux DNA was oriented to align target gene transcription with that of the lux genes . Genes in Escherichia coli and Vibrio parahaemolyticus were mutagenized, and mutants containing transposon-generated lux gene fusions produced light as a function of target gene transcription . Light production offers a simple, sensitive, in vivo indicator of gene expression. Southeast Asian J Trop Med Public Health, 1985 Mar, 16(1), 113 - 6 NAG Vibrio cholerae isolated from imported shellfish in Guam; Haddock RL et al.; A survey of imported shellfish available in public markets on the Island of Guam revealed the presence of NAG Vibrio cholerae contamination (10 of 38 sample lots positive) and high coliform counts (21 of 33 sample lots in excess of 2400 per 100 grams) . NAG V . cholerae contamination was associated with high coliform counts and origin in fresh or brackish waters rather than saltwater . Further importation of fresh shellfish from other than approved sources