Hybridization by Deoxycholate Treatment or by Biological Degradation of Bacterial Endotoxins Extracted from Smooth and Rough Strains

1970 ◽  
Vol 1 (1) ◽  
pp. 15-20
Author(s):  
L. Chedid ◽  
M. Parant ◽  
F. Parant ◽  
F. Peroux ◽  
J. J. Perez
Keyword(s):  
Sodium Deoxycholate ◽  
Salmonella Typhi ◽  
Side Chains ◽  
Biological Degradation ◽  
Specific Antibodies ◽  
Electrophoretic Migration ◽  
Bacterial Endotoxins ◽  
Phenol Water ◽  
Smooth Strain

An endotoxin extracted with phenol-water from a rough microorganism ( Salmonella typhi strain R 2 ) was hybridized with an endotoxin obtained by the Boivin technique from a smooth strain ( S. enteritidis Danysz) when they were mixed in the presence of sodium deoxycholate. These two toxic antigens could also be hybridized by incubation in citrated serum. With this new composite molecule, the presence of the hydrophilic side chains on the smooth moiety influenced the electrophoretic migration of the R antigen and greatly hindered the reactivity of the R sites with their specific antibodies.

scholarly journals HYBRID FORMATION BETWEEN BACTERIAL ENDOTOXINS

1967 ◽  
Vol 126 (1) ◽  
pp. 63-79 ◽  
Author(s):  
Jon A. Rudbach ◽  
Kelsey C. Milner ◽  
Edgar Ribi
Keyword(s):  
Bile Salt ◽  
Sodium Deoxycholate ◽  
Enteric Bacteria ◽  
Biologically Active ◽  
Antigenic Determinants ◽  
Hybrid Formation ◽  
Bacterial Endotoxins ◽  
New Type ◽  
The Individual

When endotoxins extracted from enteric bacteria were mixed in the presence of sodium deoxycholate, and the bile salt was subsequently removed by dialysis or by extraction with ethanol, a new type of endotoxin was formed. The latter material was as biologically active as the original endotoxins and possessed a combination of antigenic determinants that were previously unique to each of the individual endotoxins in the mixture. This hybrid formation between endotoxins was detected by immunodiffusion and radioautography and by quantitative precipitation procedures.

Endotoxic activity of cell-free rumen fluid from cattle fed hay or grain

1978 ◽  
Vol 24 (10) ◽  
pp. 1253-1261 ◽  
Author(s):  
T. G. Nagaraja ◽  
L. R. Fina ◽  
E. E. Bartley ◽  
H. D. Anthony
Keyword(s):  
Actinomycin D ◽  
Rumen Fluid ◽  
Quantitative Difference ◽  
Gram Negative Bacteria ◽  
Chick Embryos ◽  
Ether Extract ◽  
Gram Negative ◽  
Bacterial Endotoxins ◽  
Phenol Water ◽  
Endotoxic Activity

The cell-free rumen fluid from cattle fed hay or grain exhibited the following biological characteristics which strongly suggest the presence of endotoxin or a toxic principle similar to endotoxin of gram-negative bacteria: proved lethal to mice when injected with actinomycin D; proved extremely lethal to chick embryos: induced biphasic pyrogenic response in rabbits; enhanced susceptibility to bacterial infection in mice; evoked positive epinephrine skin reaction in rabbits and phenol–water or aqueous ether extract proved lethal to mice and chick embryos. A quantitative difference in concentrations of endotoxin was observed, based on LD50 in mice and chick embryos and response to the epinephrine skin test in rabbits. Cell-free rumen fluid of grain-fed cattle contained at least twice as much endotoxin as that of hay-fed cattle. Endotoxin in cell-free rumen fluid and in higher concentration in cattle fed grain than in those fed hay support the hypothesis that rumen bacterial endotoxins may participate in the pathogenesis of diseases associated with high grain feeding such as lactic acidosis and the sudden-death syndrome.

scholarly journals Extraction and Characterization of Lipopolysaccharide from Salmonella typhi

2021 ◽  
pp. 236-242
Author(s):  
Nilima Patel
Keyword(s):  
Carbohydrate Chain ◽  
Extraction Methods ◽  
Salmonella Typhi ◽  
Gram Negative Bacteria ◽  
Ladder Pattern ◽  
Wet Weight ◽  
Phenol Water ◽  
Phenolic Extraction ◽  
Relative Purity

There is no standard process available for high purity LPS isolation, and it may be appropriate to mix two purification steps. For Salmonella typhi LPS that were subjected to normal microbiological & biochemical screening protocols, updated phenol-water extraction protocol & non-phenolic extraction methods were also used in present research. Pellet of crude LPS obtained with wet weight was 2.0 gm. yield of LPS was more by hot phenol method 1.94 mg/ml as compared to non-phenolic method 0.40 mg/ml. relative purity of LPS obtained by hot phenol method was more as compared to non-phenolic method, as protein content was 23.60 mg/ml in LPS extracted from hot phenol method & 26.62 mg/ ml in LPS extracted from non-phenolic method. However, Nucleic acid contain was comparable in LPS extracted from both methods. Qualitative analysis showed ladder like bands of LPS extracted by hot phenol method as compared to single band obtained for LPS extracted by non-phenolic method. Findings of silver staining clearly revealed ladder pattern of multi-rung bands that are characteristics of smooth form of gram negative bacteria due to difference in length of carbohydrate chain of O-antigen component. In order to clarify diseased conditions & better work on LPS profiling, this research may be crucial to lead to more successful diagnosis and care.

scholarly journals MODIFICATION OF HOST RESPONSES TO BACTERIAL ENDOTOXINS

1965 ◽  
Vol 121 (5) ◽  
pp. 751-759 ◽  
Author(s):  
Yoon Berm Kim ◽  
Dennis W. Watson
Keyword(s):  
Standard Dose ◽  
Endotoxin Tolerance ◽  
Immunologic Tolerance ◽  
Host Responses ◽  
Immune Mechanism ◽  
Immunoelectrophoretic Analysis ◽  
Specific Antibodies ◽  
Bacterial Endotoxins ◽  
Exclusion Chromatography

Serum from rabbits rendered tolerant or immune to 100 MPD-3/kg of endotoxin when passively transferred to normal rabbits gave partial tolerance to the standard dose of endotoxin. The same serum was fractionated by DEAE chromatography into 4 major fractions. Immunoelectrophoretic analysis indicated that the 7S γ2-and the 19S γ1-immunoglobulins were separated into two distinct fractions. Of the four fractions tested, only fraction IV containing 19S γ1-immunoglobulm conferred complete pyrogenic tolerance to 100 MPD-3/kg of endotoxin. Additional fractionation of DEAE fraction IV by exclusion chromatography on sephadex G-200 gave 3 fractions. Of these only the first, containing 19S γ1-immunoglobulin conferred complete pyrogenic and lethal tolerance to normal rabbits. There was no correlation between the quantity of O-specific antibodies and the ability to transfer tolerance. It is concluded that endotoxin tolerance involves a classical immune mechanism which includes both 19S γ1-immunoglobulin specific for toxophore groups common to many endotoxins and a normally functioning RES. To avoid confusion with immunologic tolerance, it is suggested that the term endotoxin immunity be substituted for endotoxin tolerance.

The human bile salt sodium deoxycholate induces metabolic and cell envelope changes in Salmonella Typhi leading to bile resistance

2022 ◽  
Vol 71 (1) ◽  
Author(s):  
Isaac B. Olivar-Casique ◽  
Liliana Medina-Aparicio ◽  
Selena Mayo ◽  
Yitzel Gama-Martínez ◽  
Javier E. Rebollar-Flores ◽  
...  
Keyword(s):  
Bile Salt ◽  
Bile Salts ◽  
Cell Envelope ◽  
Sodium Deoxycholate ◽  
Salmonella Typhi ◽  
Metabolic Adaptation ◽  
Human Bile ◽  
Content Type ◽  
Genetic Elements ◽  
Bile Resistance

Introduction. Salmonella enterica serovar Typhi (S. Typhi) is the etiological agent of typhoid fever. To establish an infection in the human host, this pathogen must survive the presence of bile salts in the gut and gallbladder. Hypothesis. S. Typhi uses multiple genetic elements to resist the presence of human bile. Aims. To determine the genetic elements that S. Typhi utilizes to tolerate the human bile salt sodium deoxycholate. Methodology. A collection of S. Typhi mutant strains was evaluated for their ability to growth in the presence of sodium deoxycholate and ox-bile. Additionally, transcriptomic and proteomic responses elicited by sodium deoxycholate on S. Typhi cultures were also analysed. Results. Multiple transcriptional factors and some of their dependent genes involved in central metabolism, as well as in cell envelope, are required for deoxycholate resistance. Conclusion. These findings suggest that metabolic adaptation to bile is focused on enhancing energy production to sustain synthesis of cell envelope components exposed to damage by bile salts.

Investigation of a Providencia rettgeri strain that has enterobacterial common antigen in the immunogenic state

1987 ◽  
Vol 33 (12) ◽  
pp. 1055-1063
Author(s):  
Robert C. Quigley ◽  
John L. Penner
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Common Antigen ◽  
Providencia Rettgeri ◽  
E Coli ◽  
Enterobacterial Common Antigen ◽  
Antigenic Material ◽  
Phenol Water ◽  
Smooth Strain ◽  
Two Populations

Antigenic material obtained by phenol–water extraction from Providencia rettgeri strains, Escherichia coli O:14 strains, and mutants of the E. coli O:14 strain were examined by the passive (indirect) hemagglutination technique, by sodium dodecyl sulphate – polyacrylamide gel electrophoresis, and by immune blotting (lipopolysaccharide (LPS) blotting). Providencia rettgeri 965, like E. coli O;14, was demonstrated to have an enterobacterial common antigen (ECA) in the immunogenic form but, unlike E. coli O:14, it possessed characteristics of a smooth strain. Two populations of molecules were observed to occur in P. rettgeri 965 phenol–water extracts: one consisting of LPS identifiable with specific O antisera and the other of ECA molecules identifiable with E. coli O:14 antiserum or with a monoclonal antibody against ECA.

scholarly journals Structural Studies on the O-Specific Side Chains of the Cell Wall Lipopolysaccharides from Salmonella typhi and S. enteritidis.

1969 ◽  
Vol 23 ◽  
pp. 1588-1596 ◽  
Author(s):  
Carl Gustaf Hellerqvist ◽  
Bengt Lindberg ◽  
Sigfrid Svensson ◽  
Tord Holme ◽  
Alf A. Lindberg ◽  
...  
Keyword(s):  
Cell Wall ◽  
Salmonella Typhi ◽  
Side Chains ◽  
Structural Studies
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