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PubMed1976

Adhesive properties of Vibrio cholerae: nature of the interaction with isolated rabbit brush border membranes and human erythrocytes.

Nonmotile vibrio mutants lacked the ability to adhere to rabbit intestinal brush border membranes and to agglutinate human group O erythrocytes, but motile revertant vibrios isolated from such strains expressed adhesiveness equivalent to that of the original parent. Two possible explanations for the relation between vibrio motility and adhesion in these assays systems are (i) that the rate of adhesion depends on the rate of chance contact brought about by motility, and (ii) that the flagellum either acts as a carrier for the bacterial adhesin or may itself be the adhesin. The present study indicates, however, that the lack of adhesion by nonmotile vibrios did not depend on motility as such and did not involve greater rates of elution. Increasing the rate of contact between nonmotile vibrio mutants and brush border membranes by compaction did not restore the adhesive properties of the defective strains. Accordingly, we speculate that the flagellum may function in some indirect way that allows the expression of the adhesive properties, such as by acting as a carrier for a specific vibrio adhesin. Adhesion to brush borders and agglutination of human group O erythrocytes was specifically inhibited by L-fucose and various glycosides of L-fucose and to a lesser extent by D-mannose. Vibrios adhered specifically to agarose beads that carried covalently linked L-fucose on their surfaces. The results suggest that L-fucose-containing structures of eukaryotic cell surfaces may function as receptors for the vibrio adhesin and may therefore be an important determinant of host susceptibility.

Animals↗
PubMed1976

Adhesive properties of Vibrio cholerae: nature of the interaction with intact mucosal surfaces.

Two companion papers in this series have characterized the interaction between Vibrio cholerae and the surfaces of eukaryotic cells. The present paper reports studies of the association between vibrios or Salmonella enteritidis and intact slices of intestinal tissue. A significant number of differences were noted in the characteristics of bacterial adhesion in these systems. The results are interpreted to indicate the presence of at least two receptors for vibrio adhesion on the mucosal surface of the rabbit small intestine. The receptor mediating the adhesion of salmonella appeared to be distinct from these. A primary role for bacterial motility in the process of adhesion of vibrios to mucosal surfaces could not be demonstrated in the assay systems studied. Rather, loss of motility in mutant vibrios appeared to be correlated with the simultaneous loss of adhesive factors (adhesins) from the bacterial surface. The inhibition of vibrio adhesion to slices of intestinal tissues by antibody to the heat-stable antigens of V. cholerae occurred in the absence of bacterial agglutination. Agglutination in this assay system appeared to be an artifact in that it could be observed only in experiments where extremely high concentrations of vibrios were used. We speculate that such high vibrio concentrations are not likely to be present in humans at the time of infection and that agglutination in the lumen of the intestine might therefore play only a minor role in prophylactic immunity against natural cholera and other enteric infections of humans.

Agglutination↗
PubMed1978

Superoxide dismutase and catalase levels in halophilic vibrios.

Superoxide dismutase (SOD) and catalase (CAT) levels were determined for several aerobically grown halophilic vibrios and compared with those found in aerobically grown Escherichia coli K-12. The SOD levels ranged from 25 to 103.6 U/mg of protein for the vibrios compared with 44.6 U/mg of protein for E. coli. The CAT levels ranged from 2.1 to 32.1 U/mg of protein. Electrophoretic analysis of cell extracts revealed that the halophilic vibrios tested possessed only one detectable SOD enzyme, except one strain which possessed two distinct enzymes, as compared with the three SOD enzymes in aerobically grown E. coli K-12. A comparison of anaerobically and aerobically grown vibrios revealed a three- to fourfold increase in SOD activity in the aerobic cells, suggesting that oxygen acts as an inducer for SOD in the vibrios as has been reported for E. coli. In one strain, Vibrio parahaemolyticus 27519, both SOD enzymes were observed in low levels in anaerobic and at higher levels in aerobically grown cells as compared with only one SOD enzyme in anaerobically grown E. coli. This suggests that differences in SOD regulation occur between the two genera. Our results indicate that halophilic vibrios possess SOD, which could enhance viruulence by allowing the organisms to survive in oxygenated environments.

Aerobiosis↗
PubMed1976

Adhesive properties of Vibrio cholerae: adhesion to isolated rabbit brush border membranes and hemagglutinating activity.

Adhesion of vibrios to the small intestine may occur (i) by association of the bacteria with secreted mucus gel or (ii) by adherence of the bacteria to the surface of epithelial cells. In the present study, vibrios readily adhered to isolated brush border membranes obtained from rabbit intestinal epithelial cells. Adhesion was temperature dependent and required the presence of divalent cations such as calcium. The agglutination of human O erythrocytes by Vibrio cholerae was observed also, and the hemagglutination test appeared to detect the same mechanism that was involved in the adhesion of vibrios to brush borders. When the bacteria were grown in broth they were adhesive and hemagglutinating, but vibrios grown on agar plates or suspended in buffer for 15 min at 37 C lacked these abilities, even though they retained undiminished motility. These two model systems differed, however, in that strontium promoted only adhesion to brush borders. The significance of this difference remains to be determined. Vibrios were observed to penetrate intestinal mucus gel and occasionally to become entrapped in it. However, there was no evidence that vibrios attached to mucus gel.

ABO Blood-Group System↗
PubMed1978

Non-cholera vibrio infections in the United States. Clinical, epidemiologic, and laboratory features.

Non-cholera vibrios are organisms that are biochemically indistinguishable from Vibrio cholerae but do not agglutinate in vibrio 0 group 1 antiserum. Since 1972 there has been a dramatic increase in the number of these organisms referred to the Center for Disease Control for identification. Clinical, epidemiologic, and laboratory data were analyzed for 26 of 28 patients with isolates identified between January 1972 and March 1975. Thirteen (50%) of the isolates were obtained from feces of patients who had an acute diarrheal illness; no other pathogens were isolated from their feces, and all patients survived. Four (15%) patients had non-cholera vibrios isolated from other gastrointestinal or biliary tract sites; none of these patients had acute illness definitely attributable to non-cholera vibrios. Nine (35%) patients had non-cholera vibrios isolated from other tissues and body fluids; four deaths occurred in this group. Patients with acute diarrhea frequently had a history of recent shellfish ingestion or foreign travel, whereas some patients with systemic non-cholera vibrio infection had a history of recent occupational or recreational exposure to salt water.

Adolescent↗
PubMed1977

Experimental Toxigenicity of NAG Vibrios.

Some strains of NAG vibrios isolated from the stool of patients with diarrhoeal disease as well as from surface water caused an accumulation of fluid in the ligated rabbit gut loop. 5-fold concentrated sterile culture filtrates of some strains were found positive in this test as well. The volume of the accumulated fluid in gut loops injected with live cultures as well as with concentrated culture filtrates was apparently smaller than the volume accumulated after injection of non-concentrated V.cholerae culture filtrates. This points to the fact that the NAG vibrio strains belong to weaker producers of enterotoxin than the cholera vibrios. The culture filtrates of all investigated strains contained the skin toxin which was of increased vascular permeability in the skin of rabbits. Besides this, a hemorrhagic effect was found in the filtrates. The skin toxin of NAG vibrios appears to be more heat resistant than the toxin of cholera vibrios. The presence of the skin toxin in culture filtrates, however, does not correlate with the enteropathogenicity of NAG vibrio strains.

Animals↗
PubMed1976

Hydrogen-dependent organisms from the human gingival crevice resembling Vibrio succinogenes.

Twenty-eight strains of microaerophilic, motile, slightly curved gram-negative rods isolated from the gingival crevice of patients with gingivitis were studied. They seemed similar to Vibrio sputorum, though eleven strains differed in minor characters from Bergey's description under the new name Campylobacter sputorum, subspecies sputorum. The oral strains studied appeared to be closely related to several species of the genus Campylobacter and to Vibrio succinogenes. The oral strains were able to utilize gaseous hydrogen and to grow in a mineral medium with either nitrate of fumarate as hydrogen acceptor. Formate could replace hydrogen as hydrogen donor. In contrast the Campylobacter strains were not dependent on hydrogen or formate as energy source and grew poorly in mineral medium. In these nutritional and metabolic aspects the oral strains are more related to Vibrio succinogenes than to Campylobacter species. Serologically the oral strains differed from all the Campylobacter species. The GC ratio in the DNA of the oral strains varied between 48 and 50%, conform to the values described for Vibrio succinogenes. Vibrio sputorum seems a nomen conservandum and vibrio-like organisms from human infections should be tested for hydrogen-dependence before they are classified as Campylobacter species.

Agglutination Tests↗
PubMed1975

[The bacteriological diagnosis of vibrio fetus infection in man (author's transl)].

A synopsis of the present classification of Vibrio fetus is given (tab. 1, 2). Besides the presentation of the importance of these bacteria in veterinary medicine, 97 human infections have been published up to now. The geographic, age and sex distribution of these cases and the material out of which the pathogens were isolated is presented (tab. 3, 4). The clinical picture is summarized. For the culture of Vibrio fetus liver broth as described by TAROZZI incubated for at least 3-4 days was suitable as enrichment medium. The necessity of a prolonged incubation period was shown by results of continous registration of the optical density of growing Vibrio fetus cultures (fig. 1). If specimen is plated out on solid culture media before enrichment, the suitable media are chocolate agar, brain-heart infusion and Bacto-tryptose agar. For subcultures the DST-agar is also useful. Solid media cultures should be incubated microaerobically with 10% CO2 added. The resistance to Cephalothin of Vibrio fetus can be applied for selection on solid culture media. The metabolic activities of different Vibrio fetus biotypes were compared. A hitherto not reported metabolic activity, the hydrolysis of palmitic acid ester, is described (tab. 5). In the tube dilution test Vibrio fetus strains were sensitive to Gentamycin, Streptomycin, Ampicillin, Ciclacillin and Carbenicillin. They were resistent to Penicillin G, Cephalotin and Sulfonamid and showed borderline sensitivities to Tetracyclin, Kanamycin and Chloramphenicol (tab. 6).

Ampicillin↗
PubMed1977

Sensitivity of the vibrios to ultraviolet-radiation.

The ultraviolet-inactivation kinetics of a number of strains of Vibrio cholerae (classical), Vibrio cholerae (el tor), NAG vibrios and Vibrio parahaemolyticus were investigated. Statistical analyses revealed significant differences between any two of the four types of vibrio in respect to their sensitivity to U.V.

Cell Survival↗
PubMed1978

Fulminating lactose-positive Vibrio septicemia.

Recently cases of tissue invasion by as yet unnamed marine vibrios which were morphologically and biochemically similar to both Vibrio parahemolyticus and V. alginolyticus, but not identical with either of them, have been described. We have seen a patient who had serious widespread tissue infection with a halophilic, Gram-negative bacterium which was isolated from blood and leg lesions. The organism had the characteristics of the genus Vibrio, and lactose fermentation and ONPG reactions were positive. It also had a lower tolerance for sodium chloride in the nutrient broth compared with the above two vibrios. The isolate seems identical to the lactose positive (L+) Vibrio described by HOLLIS et al. (1976). Tissue infection resulting in severe necrotizing cellulitis and vasculitis was demonstrated at autopsy.

Cellulitis↗
PubMed2026

Assessment of multiple probiotic strains that protect Montipora capitata coral from infection by Vibrio coralliilyticus.

Coral disease outbreaks threaten reef ecosystems, often leading to widespread mortality and declines in coral cover. Outbreaks of tissue loss diseases like acute Montipora white syndrome (aMWS) have impacted coral populations that include the Hawaiian rice coral (Montipora capitata). Multiple strains of Vibrio coralliilyticus are known pathogens, and strain OCN008 has been demonstrated as an etiological agent of aMWS in Hawai'i. Recent work has demonstrated that probiotic bacterial strains can be used to directly treat or prevent transmission (prophylaxis) of coral diseases. Based on their production of zones of inhibition and isolation from disease-resistant corals, Pseudoalteromonas ardens R96, Pseudoalteromonas obscura P94, strain Y97 (the genomic similarity to Pseudoalteromonas piscicida is presented), Pseudoalteromonas umbrosa B95, and Vibrio tetraodonis subsp. pristinus OCN044 were assessed for their ability to impair V. coralliilyticus OCN008 infection of M. capitata during laboratory infection trials. Individual inoculation of each of the five aforementioned strains on M. capitata fragments for 48 h prior to V. coralliilyticus OCN008 inoculation resulted in up to a 93.75% reduction in mortality. These results indicate that strains of Pseudoalteromonas and Vibrio can act as prophylactics to prevent M. capitata mortality from V. coralliilyticus OCN008 infection and provide tools to improve disease resilience for Pacific corals.IMPORTANCECoral disease outbreaks are a growing threat to the continued health of coral reefs, which are already vulnerable ecosystems. Strains of the bacterium Vibrio coralliilyticus are known to infect various coral species worldwide, predominantly causing tissue loss and death of the coral animal. Previous research has indicated that constituents from healthy coral microbiomes can act as probiotics to treat or prevent coral infections, and the discovery of effective probiotics is important in the effort to further develop mitigation tools for disease outbreaks. This work provides a demonstration of probiotic species that can protect coral from tissue loss infections by a strain of Vibrio coralliilyticus and is an example of probiotics developed for coral species in Hawai'i. This work provides new tools for probiotic-based coral protection and evidence for this research as a viable avenue to protect coral in their native environments.

Animals↗
PubMed1979

Effect of chemotaxis on the interaction of cholera vibrios with intestinal mucosa.

Earlier reports from this laboratory have shown that chemotaxis is an important mechanism that expedites the in vitro association of cholera vibrios with intestinal slices and that affects the in vivo colonization and virulence of these bacteria to a significant degree. The data reported in the present communication indicate that there appears to be a chemotatic gradient attracting cholera vibrios not only to the surface of the mucus gel, but that this gradient continues for at least a considerable distance toward the base of the villi. It is shown further that a strain of Vibrio cholerae was attracted by all 20 amino acids tested, in contrast to Escherichia coli AW405 which is repelled by several of these. Finally, experiments are described that show that superior in vivo colonization of chemotatic vibrios (compared to nonchemotactic mutants) was correlated with a significantly higher degree of mucosal association. Such increased mucosal association of chemotatic vibrios has previously been shown only with mucosal slices in vitro.

Adhesiveness↗
PubMed1975

Method for detecting small numbers of Vibrio cholerae in very polluted substrates.

A method is presented for the indirect detection of Vibrio cholerae by the multiplication of two specific bacteriophages: phiH74/64 for El-Tor vibrios, and phage group IV (Mukerjee) for classical vibrios. The product to be examined is seeded in alkaline tryptone water for enrichment, as in the classical method, and is then incubated for 6 h at 37 C. Thereafter, a loopful is transferred to each of two nutrient broth (pH 9) tubes. One of these receives a drop of phage phiH74/64; the other receives a drop of phage group IV. The stock phages are diluted so as to contain about 3,800 plaque-forming units in one drop; this is the maximum amount which, when added to 10 ml of broth, will not be detected in a loopful of 1 mm diameter. The tubes containing phage phiH74/64 are incubated at 42 C; those with phage group IV are incubated at 37 C. After 18 h the cultures are killed by agitation with chloroform, and a 1-mm loopful is deposited on a layer seeded with the detector strains: Makassar 757 for El-Tor phage and V. cholerae 154 for classical cholera phage. After 4 to 5 h at 37 C, lysis appears on the spot areas if there has been phage multiplication in the respective broth tubes. With experimentally contaminated sewage water, vegetables, or stools, 1 to 10 cholera vibrios were detected in every sample. In rare cases, false-positive results were obtained by multiplication of the phage on non-cholera vibrios.

Bacteriological Techniques↗
PubMed1976

Isolation of Vibrio parahaemolyticus from fecal specimens on mannitol salt agar.

Unless laboratories use an inhibitory medium, Vibrio parahaemolyticus will be unrecognizable in fecal specimens. The use of a medium exclusively for vibrio isolation, such as thiosulfate-citrate-bile salts-sucrose agar (TCBS), however, may not be considered economically justified in the United States. The isolation and recognition of V. parahaemolyticus is reported on mannitol salt agar (MS), a medium which is used for fecal specimens here. Eight Kanagawa-positive and two of three Kanagawa-negative strains of V. parahaemolyticus grew as well on MS as on TCBS and better than on a representative enteric medium, Hektoen enteric agar (HE). Twenty-two fecal specimens from 16 noninfected individuals were inoculated with known quantities of V. parahaemolyticus, and recovery of these vibrios was assessed on TCBS, MS, and HE. Recovery of vibrios from MS and TCBS was similar when inoculum size was 10(3) colony-forming units/ml or greater. Recovery of vibrios from mixed culture was distinctly lower on HE. The colonial morphology of V. parahaemolyticus and several other bacteria on MS is illustrated.

Agar↗