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Biomedical subjects

R R Colwell

Publications and source records attributed to R R Colwell.

At least 199 records · Page 11Linked to original sources
PubMed1982

Production and characterization of monoclonal antibodies to cholera toxin.

Monoclonal antibodies against cholera toxin were produced to obtain highly specific antisera to cholera toxin. Fifteen hybridoma cell lines producing monoclonal antibodies specific for the determinants of cholera toxin were derived from the fusion of mouse myeloma cells and spleen cells from mice immunized with cholera toxin. The cell lines were stabilized, examined for specific antibody production, and immortalized by freezing cultured cells and tumor cells which had been grown subcutaneously in mice. All cell lines continued antibody secretion upon thawing. The antibodies produced by the hybridoma cell lines were characterized by determination of the class of light- and heavy-chain components and by determination of specificity for the cholera toxin subunit. All of the antibodies contained the k light chain, 4 contained the mu heavy chain, and the remaining 11 contained the gamma 1 heavy chain. Ten of the monoclonal antibodies are specific for the B subunit of cholera toxin, and three are specific for the A subunit. The remaining two appear to react with both subunits.

Animals↗
PubMed1982

Scanning electron microscope observation of the swarming phenomenon of Vibrio parahaemolyticus.

Scanning electron microscopy was used to study the production of lateral flagella and the swarming phenomenon in Vibrio parahaemolyticus. Differences in the size and diameter of the sheathed, polar flagellum and lateral flagella were apparent in these preparations. Swarming of V. parahaemolyticus was found to be similar to the swarming of Proteus spp. in that swarm cells which are heavily flagellated and elongated are formed.

Flagella↗
PubMed1982

Adsorption kinetics of laterally and polarly flagellated Vibrio.

The adsorption of laterally and polarly flagellated bacteria to chitin was measured, and from the data obtained, a modified Langmuir adsorption isotherm was derived. Results indicated that the adsorption of laterally flagellated Vibrio parahaemolyticus follows the Langmuir adsorption isotherm, a type of adsorption referred to as surface saturation kinetics, when conditions are favorable for the production of lateral flagella. When conditions were not favorable for the production of lateral flagella, bacterial adsorption did not follow the Langmuir adsorption isotherm; instead, proportional adsorption kinetics were observed. The adsorption of some polarly flagellated bacteria exhibited surface saturation kinetics. However, the binding index (the product of the number of binding sites and bacterial affinity to the surface) of polarly flagellated bacteria differed significantly from that of laterally flagellated bacteria, suggesting that polarly flagellated bacteria adsorb to chitin by a different mechanism from that used by the laterally flagellated bacteria. From the results of dual-label adsorption competition experiments, in which polarly flagellated V. cholerae competed with increasing concentrations of laterally flagellated V. parahaemolyticus, it was observed that laterally flagellated bacteria inhibited the adsorption of polarly flagellated bacteria. In contrast, polarly flagellated bacteria enhanced the adsorption of V. cholerae. In competition experiments, where V. parahaemolyticus competed against increasing concentrations of other bacteria, polarly flagellated bacteria enhanced V. parahaemolyticus adsorption significantly, whereas laterally flagellated bacteria only slightly enhanced the process. The direct correlation observed between surface saturation kinetics, the production of lateral flagella, and the ability of laterally flagellated bacteria to inhibit the adsorption of polarly flagellated bacteria suggests that lateral flagella represent a component of bacterial structure that is important in the adsorption of laterally flagellated bacteria to surfaces. A model for adsorption events of laterally flagellated bacteria is proposed, based on the evidence presented.

Adsorption↗
PubMed1982

Statistical evaluation of a quality control method for isolation of pathogenic Vibrio species on selected thiosulfate-citrate-bile salts-sucrose agars.

The recovery of Vibrio cholerae, Vibrio fluvialis, Vibrio parahaemolyticus, and Vibrio vulnificus, employing eight strains of each species, was studied by using four brands of thiosulfate-citrate-bile salts-sucrose (TCBS) agar prepared according to manufacturers' instructions and following a standardized procedure. A standardized broth inoculum of each strain was placed on duplicate plates of each brand of TCBS agar and also on tryptic soy agar (Difco Laboratories) containing 1% (wt/vol) NaCl, the latter serving as the control. Plates were inoculated in a sequence designed to compensate for bias associated with multiplication of the bacteria during the inoculation procedure. Colony counts and quality of growth were recorded after incubation for 18 h at 35 degrees C. The comparison procedure was repeated four times at weekly intervals. Data were analyzed by using an analysis of variance model. The recovery and quality of growth of each species varied significantly on the different brands of TCBS agar. Significant variability was also identified for some components of the inoculation procedure. Modifications of the inoculation procedure are suggested to minimize sources of variance. A simplified statistical procedure, based on the t test, is described for media quality control for laboratories routinely isolating pathogenic Vibrio spp.

Bacteriological Techniques↗
PubMed1982

Vibrio parahaemolyticus and related halophilic Vibrios.

Approximately 30 years have elapsed since Dr. Fujino's original discovery that Vibrio parahaemolyticus (then termed Pasteurella parahemolytica) was the cause of "summer diarrhea" in Japan. Since that finding, V. parahaemolyticus has been established as a cause of gastroenteritis in numbers and places approaching global proportions. It has been isolated in marine and estuarine areas almost worldwide and despite its halophilic nature, V. parahaemolyticus has been isolated from saline-free waters. The relationship of this organism to the environment reveals a close association with other marine organisms especially copepods on which the Vibrios depend for survival in winter months and growth in summer months. There is a uniquely provocative disparity between human strains of V. parahaemolyticus which are Kanagawa phenomenon (KP) positive and the environmental strains which to a large extent are KP negative, the significance being that pathogenicity is measured according to the Kanagawa phenomenon (hemolytic activity) reaction. The hemolysin of the pathogenic strains is a thermostable, cardiotoxic protein, which thus far has not been implicated in the mechanism(s) which causes human gastroenteritis. The interest in this organism has been widened in recent years by the finding that similar organisms, V. alginolyticus, lactose positive vibrios and group F vibrios also cause serious disease in humans.

Animals↗
PubMed1981

Bacteria associated with false-positive most-probable-number coliform test results for shellfish and estuaries.

Aerobic and facultatively anaerobic bacteria isolated from false-positive, presumptive, total coliform, most-probable-number tests of Chesapeake Bay oyster, water, and sediment samples were characterized and then classified by numerical taxonomy. A total of 538 bacterial strains clustered into 17 phena, the predominant groups of which were Enterobacteriaceae (including Escherichia coli), Aeromonas spp., and Bacillus spp. Bacillus spp. were recovered most frequently from sediment samples. Gas-producing strains which were not members of the Enterobacteriaceae were not isolated during this study. However, disproportionately large numbers of atypical and anaerogenic lactose-fermenting strains were encountered. We concluded that no single, specific bacterial group can be identified as being responsible for the false-positive reaction in the presumptive coliform test. Instead, the false-positive reaction is a result of complex interactions among various genera, representing predominantly bacteria other than coliforms.

Animals↗
PubMed1981

Effect of temperature on growth and activity of Aeromonas spp. and mixed bacterial populations in the Anacostia River.

During the winter months, total bacterial counts in the water column and in the sediment in the Anacostia River were two- to eightfold higher than at other times of the year, whereas Aeromonas spp. decreased in number of several orders of magnitude. This significant decrease in number in the Anacostia River during the cold months of the year can be explained by the low metabolic activity of Aeromonas at low temperatures.

Aeromonas↗
PubMed1981

Effects of pharmaceutical wastes on microbial populations in surface waters at the puerto rico dump site in the atlantic ocean.

A series of cruises during 1979 and 1980 to the pharmaceutical dump site located 64 km north of Arecibo, Puerto Rico, in the Atlantic Ocean, was carried out to evaluate effects of wastes on the ecology of the microflora of surface waters of the dump site. In addition to bacteriological monitoring of the waste plume created by the release of wastes from the disposal barge, stations along a series of transects, extending north from coastal waters through and beyond the dump site, were sampled. Largest numbers of culturable bacteria on marine agar were found at stations closest to shore and in the vicinity of the dump site. Bacteria recovered on marine agar were predominantly Vibrio and Aeromonas spp., with the relative abundance of these organisms decreasing as gram-positive organisms (staphylococci, micrococci, and bacilli) became dominant in areas immediately affected by waste dumping. Total numbers of bacteria (determined by acridine orange direct counts [AODC]), which were relatively stable throughout the region, and a direct estimate of viable cells (DVC), i.e., those cells responsive to additions of yeast extract and nalidixic acid, were determined by acridine orange staining and epifluorescence microscopy. Heterotrophic bacterial activity, measured by the uptake (V(max)) of C-labeled amino acids, declined relative to distance from land. Increases in specific activity indices (DVC/AODC and V(max)/AODC) were observed near the dump site. The composite results of this study, i.e., increased specific activities (determined by two methods), increased numbers of culturable marine bacteria, and marked alteration of the taxonomic composition of the culturable bacterial community in waters within and surrounding the Puerto Rico dump site, indicate demonstrable changes in the marine microbial community in the region used for waste disposal.

Journal Article↗
PubMed1981

Association of Aeromonas sobria with human infection.

Fifteen Aeromonas isolates from various human infections and nine isolates from polluted water were identified as either Aeromonas hydrophila or Aeromonas sobria and examined for cytotoxigenicity, enterotoxigenicity, adherence to epithelial cells, and other virulence-associated factors, including proteases, lipases, elastases, and hemolysins. Two groups of organisms (I and II) were distinguishable based on differences in median lethal doses in mice and cytotoxicity for Y-1 adrenal cells. Group I clinical and environmental strains had median lethal doses of less than 10(7) colony-forming units, were cytotoxic, frequently possessed several virulence-associated factors, and had lysine decarboxylase-positive or Voges-Proskauer-positive phenotypes or both. Piliation of Aeromonas was associated strongly with ability to adhere to human buccal cells, and these characteristics were associated with group I strains. Group II clinical and environmental strains had median lethal doses of greater than or equal to 10(7) colony-forming units, were not cytotoxic, and usually were lysine decarboxylase negative or Voges-Proskauer negative or both. Clinical strains in group II exhibited enterotoxigenicity, which was not detected in group II environmental strains. A sobria was more frequently associated with human infections; 13 of the 15 clinical strains were A. sobria, and 2 were A. hydrophila. On the other hand, the majority of the environmental strains (seven of nine) were A. hydrophila.

Aeromonas↗
PubMed1981

Comparison of methods for identifying Staphylococcus and Micrococcus spp.

Three methods employed to distinguish staphylococci from micrococci were compared, using clinical and environmental strains. When these methods are used, misinterpretation of results, as well as erratic results, may occur, and suggestions for eliminating these problems are provided. The most sensitive test that combines ease of use and speed in obtaining results for distinguishing the two genera is the lysostaphin susceptibility test. Two other tests, facultatively anaerobic growth in semisolid thioglycolate agar and fermentation of dextrose, may also be used to distinguish these two genera, but results are often slow in developing, are subject to technical difficulties, and may lead to incorrect assignment of certain species of staphylococci and micrococci to their proper genera.

Bacteriological Techniques↗
PubMed1980

Isolation of drug-resistant Aeromonas hydrophila from aquatic environments.

Antibiotic-resistant strains of Aeromonas hydrophila have been isolated from the natural environment in the Chesapeake Bay and areas surrounding Dacca and the Matlab region of Bangladesh. The Bangladesh strains carried resistance to chloramphenicol, streptomycin, and tetracycline, and 57% of them had a multiple streptomycin-tetracycline resistance phenotype correlated with the presence of a large plasmid. The Chesapeake Bay strains were resistant to polymyxin B ane tetracycline, but showed neither multiple resistance nor R-factor carriage. Twenty-five percent of the environmental strains were toxigenic in a Y-1 adrenal cell assay. Toxigenicity showed no positive correlation with drug resistance or with plasmid carriage. Environmental areas of heavy human impact appear to be associated with a higher incidence of antibiotic-resistant strains of aeromonads.

Aeromonas↗
PubMed1980

R-factor carriage in a group F vibrio isolated from Bangladesh.

Two group F vibrio organisms have been identified among a collection of vibrio strains isolated from the aquatic environment in Bangladesh. Neither group F strain produced a cholera-like enterotoxin. One of the isolates, BV12, contained an R plasmid conferring resistance to streptomycin and chloramphenicol.

Aeromonas↗
PubMed1980

Microbial transformation of kepone.

Pseudomonas aeruginosa strain KO3 and a mixed aerobic enrichment culture, isolated from sewage sludge lagoon water, were found to aerobically transform the chlorinated insecticide Kepone, yielding monohydro-Kepone. Identification of the product was confirmed by gas chromatography and electron impact mass spectrometry. The mixed culture and P. aeruginosa strain KO3 produced about 4 and 16%, respectively, dihydro-Kepone, determined by cochromatography using authentic standards. Reduced amounts of monohydro-Kepone, compared with the mixed and pure cultures, were produced by James River sediment microorganisms. Kepone was not utilized as a sole carbon or energy source by any of the bacteria or mixed cultures examined in this study.

Chlordecone↗
PubMed1980

Distribution and characterization of kepone-resistant bacteria in the aquatic environment.

Effects of the chlorinated insecticide Kepone on the ecology of Chesapeake Bay and James River bacteria were studied. Kepone-resistant bacteria present in a given environment were found to reflect the degree of fecal and/or high organic pollution of the sampling sites, based on total numbers and generic composition of the populations of Kepone-resistant bacteria. The presence of Kepone-resistant bacteria was found to be correlated (alpha = 0.01) with total coliforms, fecal coliforms, and total aerobic viable heterotrophic bacteria, but not with Kepone concentration, since Kepone-resistant bacteria were present in locations where Kepone could not be detected by the analytical methods used in this study. Only gram-negative bacteria, predominantly Pseudomonas, Vibrio, and Aeromonas spp., were found to be resistant to >/=10 mug of Kepone per ml. Gram-positive bacteria, i.e., Bacillus and Corynebacterium spp., were generally sensitive to >/=0.1 mug of Kepone per ml. From results of cluster analysis of taxonomic data, we determined that characteristics of Kepone-resistant bacteria included: resistance to pesticides and heavy metals; degradation of oil; positive oxidase and catalase reactions; and nitrate reduction. From results of the ecological and taxonomic analyses, we conclude that Kepone resistance in estuarine bacteria is due to the physicochemical composition of the gram-negative cell wall and not prior exposure to Kepone. Therefore, the presence of Kepone-resistant bacteria cannot serve as an indicator of Kepone contamination in the aquatic environment where gram-negative bacteria are predominant.

Aeromonas↗