Rabbit Pyrogen Research Articles

Detoxified Bacterial Endotoxins I. Preparation and Biological Properties of an Acetylated Crude Endotoxin from Salmonella typhimurium

Martin, William J. (University of Utah, Salt Lake City), and Stanley Marcus. Detoxified bacterial endotoxins. I. Preparation and biological properties of an acetylated crude endotoxin from Salmonella typhimurium. J. Bacteriol. 91:1453-1459. 1966.-Acetylation of a crude endotoxin prepared by the Roschka-Edwards (RE) procedure from a strain of Salmonella typhimurium yields a product with reduced pyrogenicity in rabbits as well as one which is nontoxic in mice. A reduction in pyrogenicity of approximately 100 times was noted with this acetylated crude endotoxin when compared with the parent RE preparation. A comparison was made of immunogenicity with mice of Boivin, RE, and acetylated Roschka-Edwards (Acet-RE) preparations with a heat-killed, phenol-preserved (HP) vaccine prepared from the same strain of S. typhimurium. Less than pyrogenic doses of all vaccines were not protective. The least pyrogenic preparation (Acet-RE) was immunogenically effective in about five times the minimal pyrogenic dose. The data suggest that the Acet-RE preparation should be considered further in the search for enteric fever vaccines with lowered potential for undesirable systemic responses.

Frequency of occurrence of native hapten among enterobacterial species.

Anacker, R. L. (Rocky Mountain Laboratory, Hamilton, Mont.), W. D. Bickel, W. T. Haskins, K. C. Milner, E. Ribi, and J. A. Rudbach. Frequency of occurrence of native hapten among enterobacterial species. J. Bacteriol. 91:1427-1433. 1966.-Smooth cultures of representative Enterobacteriaceae were screened for the presence of native hapten, a substance previously extracted with trichloroacetic acid from the protoplasmic fraction of one strain each of Escherichia coli O111:B4 and O113. Trichloroacetic acid extracts of protoplasmic fractions of the cells were analyzed for chemical composition, for constituent sugars by paper chromatography, for immunochemical relationship to endotoxin purified by gel filtration, for sedimentation behavior, and for pyrogenicity in rabbits and lethal toxicity in chick embryos. Extracts from two of three additional strains of E. coli O113, all five additional strains of E. coli O111:B4, and one strain each of E. coli O26:B6 and O55:B5 were similar to previously described native hapten in chemical composition, sedimentation properties (S(20,w), 3.7 to 5.2), biological potency (usually less than 0.1% that of corresponding endotoxin), and immunochemical relationship to endotoxin. Extracts of one strain each of E. coli O127:B8, Serratia marcescens, Citrobacter freundii, Salmonella enteritidis, and of two lipopolysaccharide-deficient mutants of S. enteritidis differed from typical native hapten. The biosynthetic relationship of native hapten to endotoxin has not yet been revealed.

STUDIES ON BACTERIAL PYROGENICITY. 3. SPECIFICITY OF THE UNITED STATES PHARMACOPEIA RABBIT PYROGEN TEST.

The specificity of the first or "presumptive" portion of the USP rabbit pyrogen test was investigated by use of a new absolute standard of reference. The reference standard was a 0.9% sodium chloride solution prepared to be pyrogen-free. Details of the preparation were described. The hypothesis was explored that the temperature response of rabbits after intravenous injection of the standard solution was independent of exogenous pyrogen. Reactions observed among the rabbits in our colony allowed a classification of these animals ranging from "consistently reliable" to "consistently unreliable." Details of the experimental results and implications for pyrogen testing are discussed. The recommendation was made that all rabbit test animals be "screened" in sham and actual tests before being used for pyrogen testing.

Studies on Bacterial Pyrogenicity1

The specificity of the first or "presumptive" portion of the USP rabbit pyrogen test was investigated by use of a new absolute standard of reference. The reference standard was a 0.9% sodium chloride solution prepared to be pyrogen-free. Details of the preparation were described. The hypothesis was explored that the temperature response of rabbits after intravenous injection of the standard solution was independent of exogenous pyrogen. Reactions observed among the rabbits in our colony allowed a classification of these animals ranging from "consistently reliable" to "consistently unreliable." Details of the experimental results and implications for pyrogen testing are discussed. The recommendation was made that all rabbit test animals be "screened" in sham and actual tests before being used for pyrogen testing.

Metabolic effects of bacterial pyrogens upon human leukocytes. I. Studies on leukocyte respiration, glycolysis, and nucleic acid synthesis.

Abstract Experiments were designed to study the effect of two pyrogens, a Pseudomonas aeruginosa polysaccharide and a Shigella toxin, upon glycolysis, respiration, and nucleic acid synthesis by normal bone marrow cells and circulating leukemic leukocytes. Both the Pseudomonas polysaccharide and Shigella toxin enhanced glycolysis and oxygen uptake by leukemic leukocytes. The Pseudomonas polysaccharide was the more effective. There was also a suggestive increase in the uptake of C 14 adenine into the ribose nucleic acid (RNA) fraction of leukemic leukocytes when high concentrations of Shigella toxin were used. But in lower concentrations Shigella toxin had no effect on the incorporation of C 14 adenine into the RNA and desoxyribose nucleic acid of rabbit bone marrow, and neither pyrogen influenced the incorporation of orotic acid into nucleic acids. From the data obtained it may be concluded that the toxic effects of bacterial pyrogens upon leukocytes devolve primarily on metabolic processes other than those concerned with nucleic acid synthesis.

Studies on bacterial pyrogenicity. I. Quantitative basis.

SummaryMultiple tests on solutions to which have been added known numbers of various bacterial species, suggest that 1000 or more organisms/ml must be present before the U.S.P. rabbit pyrogen test will yield a positive result. The potentiality is inferred of a simple culture procedure as a screen test for pyrogenicity of solutions to be employed for parenterl infusion.