Search PubMed⌕ Search

Biomedical subjects

R E Wooley

Publications and source records attributed to R E Wooley.

At least 19 recordsLinked to original sources

In vitro inhibition of Salmonella organisms isolated from reptiles by an inactivated culture of microcin-producing Escherichia coli.

OBJECTIVE: To determine whether an inactivated culture of a microcin-producing avian Escherichia coli was capable of killing Salmonella isolates from reptiles in an in vitro test system. SAMPLE POPULATION: 57 Salmonella isolate from reptiles. PROCEDURE: A wild-type avian E. coli electrotransformed with a plasmid coding for the production of microcin 24 was tested in an in vitro microassay system for its ability to kill 57 Salmonella spp isolated from reptiles. The reptile population included snakes, iguana, frilled lizards, turtles, other lizards, and unspecified reptiles. RESULTS: 44 of the Salmonella isolates were inhibited strongly, compared with the in vitro assay controls; 12 had weak inhibition, and 1 was not inhibited by the microcin-producing E. coli. Thirteen of the 57 isolates had resistance to at least 1 antibiotic, primarily streptomycin. There were 9 O serogroups identified in the 57 isolates, with serogroup H being the most prevalent (18 to 57). CONCLUSION AND CLINICAL RELEVANCE: Antibiotics are not recommended to eliminate Salmonella organisms from reptiles because of the development of antibiotic resistance. Further studies are necessary to determine whether the use of microcin-producing bacteria will be effective in controlling Salmonella infections in companion reptiles.

Animals↗

Antimicrobial effect of combinations of EDTA-Tris and amikacin or neomycin on the microorganisms associated with otitis externa in dogs.

Combinations of EDTA-Tris and two aminoglycoside antibiotics (amikacin and neomycin) were tested for synergistic activities against the microorganisms associated with otitis externa in dogs and for the solutions' stability over time. Synergistic activity was observed when EDTA-Tris plus amikacin and EDTA-Tris plus neomycin were tested against Staphylococcus intermedius, Proteus mirabilis, Pseudomonas aeruginosa, and Escherichia coli, but not against Candida albicans. Stability studies over a 3-month period indicated that the test solutions were stable at room temperature and that their antimicrobial activity was maintained.

Amikacin↗

Factors affecting endotoxin release from the cell surface of avian strains of Pasteurella multocida.

Two avian strains of Pasteurella multocida, a vaccine strain and a virulent field isolate, were investigated to determine their propensity to release endotoxin from the cell surface. Both organisms released comparable amounts of endotoxin when plasma complement proteins were present, however the virulent strain did so without the loss of viability that occurred in the vaccine strain. Blocking complement activity decreased the ability of plasma to elicit endotoxin release from the bacteria. When the cells were treated with divalent metal chelators such as trans-1, 2-diaminocyclohexane-N,N,N1,N1-tetraacetic acid (CDTA), more endotoxin was released from the vaccine strain than from the virulent isolate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of purified lipopolysaccharide (LPS) from both strains revealed virtually identical patterns. Both had patterns considered typical of rough LPS. Challenge studies in 8 weeks old turkeys showed that the field strain induced endotoxemia of longer duration than the vaccine strain and produced greater mortality.

Animals↗

A survey of potential virulence markers from avian strains of Pasteurella multocida.

Twenty-four isolates of Pasteurella multocida from clinical cases of fowl cholera and the Clemson University vaccine strain were surveyed for the presence of potential virulence markers. Membrane proteins, enzymatic activity of the membrane proteins, and carbohydrate fermentation patterns were also determined to demonstrate phenotypic relationships within the groups. Few differences were found in these phenotypic characteristics among the isolates. Almost all the organisms produced siderophore and were hemolytic on turkey red blood cells. No extracellular enzyme or bacteriocin activity was detected and little antibiotic resistance was found. However, many organisms contained plasmids and demonstrated some degree of resistance to complement. Both characteristics were correlative markers in Pasteurella multocida isolated from birds with fowl cholera.

Animals↗

Comparison of phenotypic characteristics of Salmonella spp isolated from healthy and ill (infected) chickens.

Phenotypic characteristics of 12 paired, Salmonella serotypes isolated from healthy and ill chickens were compared. Variables compared included antibiotic resistance profiles, production of colicins and siderophores, mannose-sensitive hemagglutination of erythrocytes, resistance to serum complement, carbon source utilization, presence and transmissibility of R plasmids, and invasiveness in primary chicken kidney cell culture. Differences were found between pairs for utilization of carbon sources, mannose-sensitive hemagglutination of erythrocytes, and invasiveness in cell culture.

Animals↗

Microflora associated with the skin of the bowhead whale (Balaena mysticetus).

A study of the microbiological flora isolated from cultures of normal and lesional skin tissue samples collected from 19 bowhead whales (Balaena mysticetus) over a 4 yr period is presented. These cultures were obtained from 30 tissue samples (17 normal, 13 lesion) and 248 swab samples (157 normal, 91 lesion). Seven hundred-thirty bacterial and yeast isolations were made (285 normal, 445 lesion). Distribution revealed that 56% of the gram positive bacterial isolates, 75% of the gram negative bacterial isolates and 64% of the yeast isolates recovered were associated with lesional skin. It was found that 80% of one group of Corynebacterium sp. isolates, 90% of the Acinetobacter sp. isolates and 94% of the Moraxella sp. isolates were associated with lesional skin. Although the primary yeasts recovered were Candida spp., they were found on both normal and lesional skin. Enzymatic assays of isolates from normal and lesional skin demonstrated production of enzymes capable of causing necrosis. The majority of the microorganisms recovered were facultative anaerobes and many of them could be considered potential pathogens of mammalian hosts.

Animals↗

Survival of coliforms and bacterial pathogens within protozoa during chlorination.

The susceptibility of coliform bacteria and bacterial pathogens to free chlorine residuals was determined before and after incubation with amoebae and ciliate protozoa. Viability of bacteria was quantified to determine their resistance to free chlorine residuals when ingested by laboratory strains of Acanthamoeba castellanii and Tetrahymena pyriformis. Cocultures of bacteria and protozoa were incubated to facilitate ingestion of the bacteria and then were chlorinated, neutralized, and sonicated to release intracellular bacteria. Qualitative susceptibility of protozoan strains to free chlorine was also assessed. Protozoa were shown to survive and grow after exposure to levels of free chlorine residuals that killed free-living bacteria. Ingested coliforms Escherichia coli, Citrobacter freundii, Enterobacter agglomerans, Enterobacter cloacae, Klebsiella pneumoniae, and Klebsiella oxytoca and bacterial pathogens Salmonella typhimurium, Yersinia enterocolitica, Shigella sonnei, Legionella gormanii, and Campylobacter jejuni had increased resistance to free chlorine residuals. Bacteria could be cultured from within treated protozoans well after the time required for 99% inactivation of free-living cells. All bacterial pathogens were greater than 50-fold more resistant to free chlorine when ingested by T. pyriformis. Escherichia coli ingested by a Cyclidium sp., a ciliate isolated from a drinking water reservoir, were also shown to be more resistant to free chlorine. The mechanism that increased resistance appeared to be survival within protozoan cells. This study indicates that bacteria can survive ingestion by protozoa. This bacterium-protozoan association provides bacteria with increased resistance to free chlorine residuals which can lead to persistence of bacteria in chlorine-treated water. We propose that resistance to digestion by predatory protozoa was an evolutionary precursor of pathogenicity in bacteria and that today it is a mechanism for survival of fastidious bacteria in dilute and inhospitable aquatic environments.

Animals↗

Comparison of virulence factors and R plasmids of Salmonella spp. isolated from healthy and ill swine.

The antibiotic resistance and virulence profiles of Salmonella spp. isolated from healthy (group 1) and ill (group 2) swine were compared. Parameters studied included colicin and siderophore production; mannose-sensitive hemagglutination of erythrocytes; resistance to the lethal effect of serum complement; resistance to antibiotics; and the transmissibility of these characteristics to recipient organisms. Group 1 (19 isolates) had 14 serotypes, and group 2 (20 isolates) had 2 serotypes. Isolates from group 2 were resistant to more antibiotics and had a greater ability to hemagglutinate erythrocytes and transfer R plasmids to recipient organisms, but a lesser ability to produce siderophore than group 1. All 39 isolates resisted the lethal effects of serum complement. Colicin was produced by 1 of 19 from group 1 and 0 of 20 from group 2. A donor Escherichia coli isolated from a pig with enteritis transferred R plasmids to 62% of group 1 and 0% of group 2 Salmonella spp. when they were used as recipient organisms. A transconjugant from the mating of donor E. coli to a group 1 Salmonella spp. was further able to pass an R plasmid to recipient E. coli and salmonellae. Plasmid isolation from group 1 yielded 1 of 19 strains with a 56-megadalton plasmid, while 20 of 20 strains from group 2 contained three to five plasmids from 2.4 to 60 megadaltons in size.

Animals↗

Effect of EDTA-tris on an Escherichia coli isolate containing R plasmids.

Solutions of ethylenediaminetetraacetate (EDTA)-tris combined with antibiotics have been shown to be effective in treating selected cases of persistent bacterial infections. Basic techniques in microbial genetics, including mating frequencies, chemical elimination of R plasmids, isolation of plasmid DNA and agarose gel electrophoresis, were used to determine if EDTA-tris has a curing effect on an R plasmid as part of its clinical action. Results of this study indicated that EDTA-tris by itself eliminated an antibiotic resistance marker from a clinical isolate of Escherichia coli and when combined with another chemical curing agent altered the isolate's mating frequency.

Conjugation, Genetic↗

Uptake of antibodies in gram-negative bacteria exposed to EDTA-Tris.

Ethylene diamine tetra-acetic acid disodium salt (3.22 mM) and Tris (hydroxymethyl aminomethane) (50 mM) was found to cause potentiation of antibiotic actions and/or increased uptake of antibiotics when assessed by bacterial inhibition kinetic studies and uptake of radiolabeled antibiotics. Solutions of EDTA-Tris potentiate the effect of tetracycline, dihydrostreptomycin and penicillin when reacted with Escherichia coli. A similar effect was observed for tetracycline when incubated with Proteus vulgaris and penicillin, chloramphenicol, dihydrostreptomycin and tetracycline when incubated with Pseudomonas aeruginosa. These effects were noted by decreasing bacterial counts in the bacterial inhibition kinetic studies and increased uptake of radiolabeled antibiotics.

Animals↗

Antimicrobic effects of Lactobacillus fermentation on edible waste material contaminated with infected carcasses.

Survival of Newcastle disease virus (NDV) and Salmonella enteritidis serovar typhimurium (introduced as infected carcasses) in Lactobacillus-fermented edible waste material was studied to determine the ability of the fermentation to activate disease agents in carcasses. Two trials were conducted. In the 1st trial, the fermented wastes contained 20% infected carcasses (protein) consisting of equal numbers of chicken (NDV) and rat (Salmonella) carcasses, and the 2nd trial contained 40% carcasses (protein). Mixtures were incubated at 20 C, 30 C, and 40 C for 216 hours. Samples were obtained daily for quantitative virus and bacterial isolation. Temperature, pH, and redox potential were monitored. In both trials, pH and redox potential changes occurred between 24 and 48 hours depending on the incubation temperature. In both trials, NDV survived 4 days at 20 C, 2 days at 30 C, and 1 day at 40 C. Salmonella enteritidis serovar typhimurium survived 5 days at 20 C, 1 day at 30 C, and was not isolated from the 40 C samples after 24 hours in both trials.

Animal Feed↗

Action of EDTA-Tris and antimicrobial agent combinations on selected pathogenic bacteria.

The concentration of EDTA-Tris (3.22 mM EDTA and 0.05 M Tris) used as a lavage to treat otitis externa, cystitis or other persistent infections in dogs and cats, was found to prevent the growth of Pseudomonas aeruginosa, Staphylococcus aureus and beta streptococci when present in growth media. Pseudomonas aeruginosa was rapidly lysed in this solution. Escherichia coli and Proteus vulgaris grew in the presence of EDTA-Tris, but to a lesser extent than the controls without these compounds. Minimal inhibitory concentrations (MIC) for 8 antimicrobial agents with and without EDTA-Tris were determined for E. coli and P. vulgaris. A potentiation effect on E. coli (greater than 50% decrease MIC) was observed when EDTA-Tris was combined with penicillin, oxytetracycline or chloramphenicol. A similar effect was observed with P. vulgaris when combinations of EDTA-Tris plus gentamicin, oxytetracycline, polymyxin-B or triple sulfa were used. The results of this study indicate that EDTA-Tris appears to have merit in selected cases of otitis externa, cystitis or other persistent infections where lavage might be used.

Animals↗

Viricidal effects of Lactobacillus and yeast fermentation.

The survival of selected viruses in Lactobacillus- and yeast-fermented edible waste material was studied to determine the feasibility of using this material as a livestock feed ingredient. Five viruses, including Newcastle disease virus, infectious canine hepatitis virus, a porcine picornavirus, frog virus 3, and bovine virus diarrhea, were inoculated into a mixture of ground food waste (collected from a school lunch program) containing Lactobacillus acidophilus. Mixtures were incubated at 20, 30, and 40 degrees C for 216 h. In a second trial, four viruses, including Newcastle disease virus, infectious canine hepatitis virus, frog virus 3, and a porcine picornavirus, were inoculated into similar edible waste material containing Saccharomyces cerevisiae. Mixtures were incubated at 20 and 30 degrees C for 216 h. Samples were obtained daily for quantitative (trial 1) and qualitative (trial 2) virus isolation. Temperature, pH, and redox potential were monitored. Controlled pH and temperature studies were also done and compared with the inactivation rates in the fermentation processes. In trial 1 (Lactobacillus fermentation), infectious canine hepatitis virus survived the entire test period in the fermentation process but was inactivated below pH 4.5 in the controlled studies. Newcastle disease virus was inactivated by day 8 in the fermentation process and appeared to be primarily heat sensitive and secondarily pH sensitive in the controlled studies. The porcine picornavirus survived the fermentation process for 8 days at 20 degrees C but was inactivated more rapidly at 30 and 40 degrees C. The controlled studies verified these findings. Frog virus 3 was inactivated by day 3 in the fermentation process and appeared to be sensitive to low pH in the controlled studies. Bovine virus diarrhea was rapidly inactivated in the fermentation process (less than 2 h) and was pH and temperature sensitive. In trial 2 (yeast fermentation), infectious hepatitis virus survived the entire test period in the fermentation process. Newcastle disease virus was inactivated by day 7 at 20 degrees C and day 6 at 30 degrees C. The porcine picornavirus was inactivated by day 7 at 30 degrees C but survived the entire test period at 20 degrees C. Frog virus 3 was inactivated by day 3 at 20 degrees C and day 2 at 30 degrees C.

Fermentation↗

In vitro action of combinations of antimicrobial agents and EDTA-tromethamine on Escherichia coli.

Combinations of EDTA-tromethamine and 7 antimicrobial agents [chloramphenicol, nalidixic acid, oxytetracycline, penicillin, polymyxin B, streptomycin, and triple sulfa (sulfamethazine, sulfapyridine, and sulfathiazole]) were tested for synergistic activities against Escherichia coli. Three in vitro tests were used including minimal inhibitory concentrations of the drugs, a 2-dimensional microtiter checkerboard technique, and bacterial inhibition studies. A strong synergistic inhibitory action was observed with combinations of EDTA-tromethamine plus penicillin and EDTA-tromethamine plus oxytetracycline. Combinations of EDTA-tromethamine plus chloramphenicol also had a synergistic action, but to a lesser extent than that with penicillin and oxytetracycline. When streptomycin or nalidixic acid was mixed with EDTA-tromethamine, a slight synergistic action was noticed. There was no synergistic effect recorded with combinations of EDTA-tromethamine and triple sulfa. Combinations of EDTA-tromethamine and polymyxin B had an antagonistic effect.

Animals↗

In vitro action of combinations of antimicrobial agents and EDTA-tromethamine on Pseudomonas aeruginosa.

Combinations of EDTA-tromethamine and 7 antimicrobial agents (chloramphenicol, nalidixic acid, oxytetracycline, penicillin, polymyxin-B, streptomycin, and triple sulfa) were tested for synergistic activities against Pseudomonas aeruginosa. Three in vitro tests were used, including minimal inhibitory concentrations of the drugs, a 2-dimensional Microtiter checkerboard technique, and bacterial inhibition studies. A synergistic inhibitory action was observed with combinations of EDTA-tromethamine plus penicillin and EDTA-tromethamine plus oxytetracycline. When chloramphenicol, streptomycin, nalidixic acid, polymyxin-B, or triple sulfa was mixed with EDTA-tromethamine, synergistic action did not occur.

Animals↗

In vitro effect of combinations of antimicrobial agents and EDTA-tromethamine on certain gram-positive bacteria.

Combinations of EDTA-tromethamine and 7 antimicrobial agents (chloramphenicol, nalidixic acid, oxytetracycline, penicillin, polymyxin-B, streptomycin, and a triple sulfonamide preparation) were tested for synergistic activities against Staphylococcus aureus, Corynebacterium renal, Listeria monocytogenes, Erysipelothrix rhusiopathiae, and a beta-hemolytic streptococci. Two in vitro tests were used--minimal inhibitory concentrations of the drugs and a 2-dimensional Microtiter checkerboard technique. A slight synergistic action was seen when L monocytogenes was exposed to combinations of penicillin-EDTA or streptomycin-EDTA.

Anti-Bacterial Agents↗