Search PubMed⌕ Search

Biomedical subjects

R J Seidler

Publications and source records attributed to R J Seidler.

At least 37 records · Page 2Linked to original sources

Rapid microimmunodiffusion method with species-specific antiserum raised to purified antigen for identification of Vibrio vulnificus.

An antigen common to Vibrio vulnificus strains, designated VVA, was purified by ammonium sulfate precipitation, gel filtration, ion-exchange column chromatography, and preparative gel electrophoresis. The molecular weight of VVA was 64,000 when estimated by gel filtration and 40,000 when measured by denaturing polyacrylamide gel electrophoresis. Antiserum prepared against purified VVA (anti-VVA serum) did not agglutinate whole cells of V. vulnificus. Therefore, VVA was considered a possible internal antigen. By using anti-VVA serum, a microimmunodiffusion method was designed to detect the antigen VVA in bacterial cell lysates prepared from a single colony. This simple method allowed the specific identification of V. vulnificus as soon as 10 h after antigen preparation and therefore can be a useful tool in the identification of V. vulnificus from environmental or clinical specimens. VVA was not detected as a line of complete identity in some 20 other Vibrio species or in 7 other bacterial genera. VVA was present in all 63 isolates of V. vulnificus obtained from clinical and nonclinical sources.

Antibodies, Bacterial↗

Demonstration of a common antigen in sonicated cells for identification of Vibrio vulnificus serotypes.

Ouchterlony immunodiffusion of sonicated Vibrio vulnificus cells illustrated a single major precipitation line with antiserum prepared from whole cells of the same species. Antigenic analysis by two-dimensional immunoelectrophoresis verified the presence of a single dominant precipitation line. Tandem two-dimensional immunoelectrophoretic analyses of V. vulnificus antigens from various strains revealed one fused precipitation line of identity. No fused precipitation lines were seen with other Vibrio species tested. This dominant antigen, designated VVA, was not dialyzable, lost antigenicity by heating at 100 degrees C but not at 70 degrees C, and was precipitated by 70%, but not by 50%, saturated ammonium sulfate. VVA was not present in concentrated (20X) spent culture medium. VVA is possibly an intracellular protein specific to V. vulnificus species and may be useful in the serological identification of this important human pathogen.

Antigens, Bacterial↗

Nucleic acids of species of Lactobacillus.

Nucleic acid composition of 17 cultures representing the type or neotype strains of 15 named Lactobacillus species was studied. Nucleic acid characterization of these isolates was accompanied by a comparative study of conventional phenotypic reactions. The overall guanine plus cytosine mean deoxyribonucleic acid base composition ranged from 33 to 50% and genome sizes varied between 700 and 1500 X 10(6) daltons. Sporolactobacillus inulinus contained 2500 X 10(6) daltons of chromosomal deoxyribonucleic acid and was therefore similar in size to members of the genus Bacillus. Deoxyribonucleic acid hybridization at a temperature 15 degrees C below the thermal melting temperature confirmed the extreme molecular heterogeneity of many species. The genus can be divided into three major groups, each containing four or more species based on a combination of nucleic acid characteristics and conventional phenotypic reactions. Group I (33 to 39% guanine plus cytosine) contains Lactobacillus jensenii, Lactobacillus helveticus, Lactobacillus helveticus nov. ssp. jugurt, Lactobacillus acidophilus, Lactobacillus salivarius, and Lactobacillus sanfrancisco. Group II (42 to 48% guanine plus cytosine contains Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus brevis, and Group III (48 to 50% guanine plus cytosine) contains Lactobacillus delbrueckii, Lactobacillus delbrueckii nov. ssp. bulgaricus, Lactobacillus delbrueckii nov. ssp. lactis, Lactobacillus delbrueckii nov. ssp. leichmanii, Lactobacillus fermentum, and Lactobacillus fermentum nov. ssp. cellobiosis.

Base Composition↗

Association of metal tolerance with multiple antibiotic resistance of bacteria isolated from drinking water.

Bacterial isolates from the drinking water system of an Oregon coastal community were examined to assess the association of metal tolerance with multiple antibiotic resistance. Positive correlations between tolerance to high levels of Cu2+, Pb2+, and Zn2+ and multiple antibiotic resistance were noted among bacteria from distribution waters but not among bacteria from raw waters. Tolerances to higher levels of Al3+ and Sn2+ were demonstrated more often by raw water isolates which were not typically multiple antibiotic resistant. A similar incidence of tolerance to Cd2+ was demonstrated by isolates of both water types and was not associated with multiple antibiotic resistance. These results suggest that simultaneous selection phenomena occurred in distribution water for bacteria which exhibited unique patterns of tolerance to Cu2+, Pb2+, and Zn2+ and antibiotic resistance.

Anti-Bacterial Agents↗

Medium-dependent production of extracellular enterotoxins by non-O-1 Vibrio cholerae, Vibrio mimicus, and Vibrio fluvialis.

Fluid accumulation at 4 h in the intestines of suckling mice enabled us to distinguish non-O-1 Vibrio cholerae, V. mimicus, and V. fluvialis clinical isolates from environmental isolates. Enterotoxin production was culture medium dependent. Filtrates of cultures grown in tryptic soy broth without glucose but with added 0.5% NaCl did not exhibit marked enterotoxin activity in the assay. Culture filtrates of all clinical strains grown in brain heart infusion broth supplemented with 0.5% NaCl induced large amounts of fluid accumulation in mouse intestines. However, most environmental strains grown in brain heart infusion broth amended as described above were unable to induce fluid accumulation. The enterotoxin present in culture filtrates lost activity at 56 degrees C and appeared to be distinct from previously described virulence factors, including the well-described cholera toxin. The new enterotoxin could represent an important virulence mechanism common to all three species.

Culture Media↗

Legionella incidence and density in potable drinking water supplies.

The incidence and density of Legionella spp. in raw water, water at various stages of treatment, and in potable distribution water were determined by direct immunofluorescence. The number of cells reacting with Legionella-specific fluorescent antibody conjugates in raw waters ranged from about 10(4) to 10(5) cells/liter, whereas the concentrations of fluorescent antibody-positive cells in the distribution waters were generally 10- to 100-fold lower than in the raw source waters. No viable or virulent Legionella strains were isolated from either the source or distribution waters. However, Legionella spp. are infrequently isolated from water at temperatures below 15 degrees C as was the case in the system surveyed in this study.

Fluorescent Antibody Technique↗

Broth medium for enrichment of Vibrio fluvialis from the environment.

A medium was designed for the enrichment and enumeration of Vibrio fluvialis from environmental samples. The medium contains 1% peptone plus 4% sodium chloride and 5 micrograms of novobiocin per ml, pH 8.5. This V. fluvialis enrichment medium (FEM) was tested, in comparison with alkaline peptone (AP), in field samplings. A total of 177 samples (estuarine waters and sediment, sewage, and crabs) collected over a 14-month period were examined with FEM and with AP broth. Results showed that FEM was more effective than AP in detecting V. fluvialis, particularly from water and sewage samples with low salinities (less than 6%). The best recovery of V. fluvialis occurred when both enrichment media were used simultaneously.

Brachyura↗

Vibrio factors cause rapid fluid accumulation in suckling mice.

Non-O-1 and O-1 Vibrio cholerae and Vibrio fluvialis isolated from clinical and environmental sources were examined for virulence factor production in 3-day-old suckling mice and in Y-1 tissue culture. The responses of the suckling mice to intragastrically administered bacterial cultures were measured by intestinal fluid accumulation (FA), diarrhea, and mortality. Regardless of the O-serovar, source of isolation, or ability to produce cholera toxin, all strains of V. cholerae stimulated increased FA, which was measurable in the mice at 4 h post-inoculation. The factor(s) causing these symptoms was found to be distinct from cholera toxin by the kinetics of FA and serological difference from cholera toxin based on in vivo neutralization tests. In most instances, FA was followed by high rates of mortality. Y-1 mouse adrenal tumor cell assays also showed that many V. cholerae produced extracellular heat-labile cytotoxic factor(s), and many cholera toxin-negative strains also caused a cytotonic-like morphological response. The majority of V. fluvialis strains produced smaller amounts of cytotoxic factor(s) but no cytotoxic reactions. The factor which stimulates rapid FA in suckling mice could be one of several virulence-associated factors contributing to diarrheal disease by nontoxigenic vibrios, but this is not verified at present.

Adrenal Gland Neoplasms↗

Vibrio furnissii (formerly aerogenic biogroup of Vibrio fluvialis), a new species isolated from human feces and the environment.

Strains formerly classified as the aerogenic (gas-producing) biogroup of Vibrio fluvialis were shown by DNA relatedness to be a separate species. The species was named Vibrio furnissii sp. nov. (type strain ATCC 35016 = CDC B3215). Three strains of V. furnissii were 79% or more related to the type strain of V. furnissii and about 50% related to the type strain of V. fluvialis. V. fluvialis strains were 40 to 64% related to the type strain of V. furnissii. Divergence in related sequences was only 0.0 to 1.5% among strains of V. furnissii and among strains of V. fluvialis but was 5.0 to 8.0% in interspecific reactions between V. fluvialis and V. furnissii. V. furnissii was aerogenic (produced gas from the fermentation of carbohydrates), whereas V. fluvialis was anaerogenic (did not produce gas from the fermentation of carbohydrates). Another test of some help in differentiating the two species was fermentation of L-rhamnose (57% positive for V. furnissii and negative for V. fluvialis). In addition to the reactions above, V. furnissii is distinguished from other salt-requiring vibrios on the basis of its positive reactions in tests for Møller L-arginine, L-arabinose, maltose, and D-mannitol and its negative reactions for Møller L-lysine and L-ornithine, lactose, and Voges-Proskauer. V. furnissii has been isolated from patients with acute gastroenteritis in at least two outbreaks of food poisoning; its role as a cause of diarrhea needs further study.

Acute Disease↗

Selection of antibiotic-resistant standard plate count bacteria during water treatment.

Standard plate count (SPC) bacteria were isolated from a drinking-water treatment facility and from the river supplying the facility. All isolates were identified and tested for their resistance to six antibiotics to determine if drug-resistant bacteria were selected for as a consequence of water treatment. Among the isolates surviving our test procedures, there was a significant selection (P less than 0.05) of gram-negative SPC organisms resistant to two or more of the test antibiotics. These bacteria were isolated from the flash mix tank, where chlorine, alum, and lime are added to the water. Streptomycin resistance in particular was more frequent in this population as compared with bacteria in the untreated river water (P less than 0.01). SPC bacteria from the clear well, which is a tank holding the finished drinking water at the treatment facility, were also more frequently antibiotic resistant than were the respective river water populations. When 15.8 and 18.2% of the river water bacteria were multiply antibiotic resistant, 57.1 and 43.5%, respectively, of the SPC bacteria in the clear well were multiply antibiotic resistant. Selection for bacteria exhibiting resistance to streptomycin was achieved by chlorinating river water in the laboratory. We concluded that the selective factors operating in the aquatic environment of a water treatment facility can act to increase the proportion of antibiotic-resistant members of the SPC bacterial population in treated drinking water.

Anti-Bacterial Agents↗

Vibrio vulnificus biogroup 2: new biogroup pathogenic for eels.

Clinical and nonclinical isolates of the lactose-positive Vibrio vulnificus were compared with Vibrio strains isolated from lesions on eels (Anguilla japonica) cultured commercially in Japan. Strains were compared phenotypically and antigenically, for pathogenicity to mice and eels, and for genetic relatedness. The strains isolated from diseased eels differed phenotypically from the original species description of V. vulnificus in that they were negative for indole production, ornithine decarboxylase activity, growth at 42 degrees C, and acid production from mannitol and sorbitol. No relationship between the surface antigens of V. vulnificus strains from environmental and clinical sources and the strains from diseased eels was observed. Typical V. vulnificus strains and the eel isolates were pathogenic to mice; however, only those strains originally isolated from diseased eels were found to be pathogenic to eels. Results of DNA-DNA competition experiments revealed that there was greater than 90% relative reassociation between clinical and nonclinical V. vulnificus and strains from diseased eels. Based on the results of the DNA-DNA competition experiments, we conclude that the strains isolated from diseased eels were V. vulnificus; however, the differences in phenotypic characteristics and eel pathogenicity indicated that these strains represent a different biogroup. Therefore, we propose that strains phenotypically similar to the type strain of the species (ATCC 27562) be classified as V. vulnificus biogroup 1 and the strains phenotypically similar to those isolated from diseased eels be classified as V. vulnificus biogroup 2 represented by the reference strain ATCC 33148.

Animals↗

Failure of the most-probable-number technique to detect coliforms in drinking water and raw water supplies.

A procedure was developed to detect false-negative reactions (interference) in the standard most-probable-number (S-MPN) technique for coliform enumeration of untreated surface water and potable water supplies. This modified MPN (M-MPN) procedure allowed a quantitative assessment of the interference with coliform detection in untreated surface water and potable water supplies. Coliform interference was found to occur in the presumptive, confirmed, and completed tests of the S-MPN technique. When coliforms were present, interference with their detection occurred in over 80% of the samples. The inferior nature of the S-MPN was revealed by the 100% increase in the incidence of completed coliform-positive drinking water samples obtained with the M-MPN technique. The M-MPN procedure was also superior to the standard membrane filter technique. Eight different species of coliforms were recovered from false-negative tests, including Citrobacter, Enterobacter, Klebsiella, and Escherichia coli (in decreasing order of occurrence). The use of standard MPN techniques for monitoring potable water supplies may lead to a false security that the drinking water supply is potable, i.e., free from indicator bacteria.

Bacteriological Techniques↗

Coliform species recovered from untreated surface water and drinking water by the membrane filter, standard, and modified most-probable-number techniques.

The species of total coliform bacteria isolated from drinking water and untreated surface water by the membrane filter (MF), the standard most-probable-number (S-MPN), and modified most-probable-number (M-MPN) techniques were compared. Each coliform detection technique selected for a different profile of coliform species from both types of water samples. The MF technique indicated that Citrobacter freundii was the most common coliform species in water samples. However, the fermentation tube techniques displayed selectivity towards the isolation of Escherichia coli and Klebsiella. The M-MPN technique selected for more C. freundii and Enterobacter spp. from untreated surface water samples and for more Enterobacter and Klebsiella spp. from drinking water samples than did the S-MPN technique. The lack of agreement between the number of coliforms detected in a water sample by the S-MPN, M-MPN, and MF techniques was a result of the selection for different coliform species by the various techniques.

Bacteriological Techniques↗

Impact of verification media and resuscitation on accuracy of the membrane filter total coliform enumeration technique.

Verification of membrane filter total coliform colonies was compared in lauryl tryptose broth, and m-LAC broth primary media and brilliant green-lactose-bile broth and EC broth secondary media. Verification in m-LAC broth yielded the greatest number of aerogenic isolates for both untreated surface water and drinking water samples. Verification in brilliant green-lactose-bile broth increased the number of false-negative reactions. At least 90% of the isolates aerogenic in primary verification media and anaerogenic in brilliant green-lactose-bile broth were representative of the coliform genera. The addition of a resuscitation step in the membrane filter technique did not yield greater numbers of verified coliforms per sample. Verification of both typical and atypical colonies in m-LAC broth resulted in a 10-fold increase in coliform numbers from untreated surface water. With drinking water, verification of both colony types resulted in an increase from less than 1 coliform per 100 ml to greater than 1/100 ml. A single-step verification in m-LAC broth is proposed as a more rapid and sensitive coliform verification procedure than the standard technique.

Aerobiosis↗