scholarly journals Protection of rabbits against experimental pasteurellosis by vaccination with a potassium thiocyanate extract of Pasteurella multocida.

1985 ◽  
Vol 49 (3) ◽  
pp. 498-504 ◽  
Author(s):  
D H Ringler ◽  
G K Peter ◽  
C E Chrisp ◽  
D F Keren
Keyword(s):  
Pasteurella Multocida ◽  
Potassium Thiocyanate

Enhancement of respiratory immunity to Pasteurella mu/tocida by cholera toxin in rabbits

1996 ◽  
Vol 30 (2) ◽  
pp. 120-126 ◽  
Author(s):  
M. A. Suckow ◽  
T. L. Bowersock ◽  
K. Nielsen ◽  
C. F. Grigdesby
Keyword(s):  
Cholera Toxin ◽  
Pasteurella Multocida ◽  
Potassium Thiocyanate ◽  
Nasal Lavage ◽  
Antibody Activity ◽  
Serum Samples ◽  
Activity Groups ◽  
Numeric Scale ◽  
Phosphate Buffered Saline ◽  
Lesion Severity

Cholera toxin (CT) is a potent adjuvant for the mucosal immune system. The purpose of this study was to determine if coadministration of CT with a potassium thiocyanate extract of Pasteurella multocida (PTE) leads to enhanced anti-PTE antibody activity and increased protection of rabbits against infection with P. multocida and associated disease. Groups of rabbits were immunized intranasally on days 0, 7, and 14, with phosphate buffered saline (PBS), 200 µg of CT, 1.0 mg of PTE, or 1.0 mg PTE with 200 ±g CT. Nasal lavage and serum samples were collected over 28 days after initial immunization and evaluated by ELISA for specific antibody directed against PTE. Marked increases in serum (IgG) and nasal lavage (IgA) anti-PTE antibody activity were found beginning after day 14 in rabbits immunized with PTE. Rabbits immunized with PTE and CT demonstrated further increases in this activity. Tracheobronchial lavage samples collected at the time of necropsy demonstrated a significant level of anti-PTE IgA activity in animals immunized with PTE, and coadministration with CT stimulated a further significant increase in this activity. Groups of similarly immunized rabbits were challenged 16 days after initial immunization with 5 × 107 CFUs of P. multocida. Nasal lavage samples were cultured for P. multocida over the next 10 days. Rabbits were euthanized within 10 days after challenge, tissues cultured for P. multocida, and histopathologic lesion severity graded using a numeric scale. Rabbits immunized with PTE survived longer, had less severe lesions of the lungs, pleura, and liver, and fewer P. multocida CFUs cultured from samples than PBS or CT controls. Coadministration of CT led to further reductions in lesion severity of those tissues and numbers of P. multocida CFUs cultured from samples. Increased nasal turbinate atrophy of rabbits immunized with PTE with or without CT was associated with increased mean survival time. In summary, coadministration of CT with PTE enhanced protective immunity to P. multocida disease and infection in rabbits.

Antigenic analysis of Pasteurella multocida (serotype 1) by crossed immunoelectrophoresis: characterization of whole cell associated antigens

1980 ◽  
Vol 26 (12) ◽  
pp. 1392-1402
Author(s):  
J. L. Bhasin ◽  
L. Lapointe-Shaw
Keyword(s):  
Pasteurella Multocida ◽  
Cell Envelope ◽  
Potassium Thiocyanate ◽  
Antigenic Analysis ◽  
Whole Cell ◽  
Heat Stable ◽  
Crossed Immunoelectrophoresis ◽  
Serotype 1 ◽  
Reference Preparation ◽  
Antigen Antibody

Crossed immunoelectrophoresis and other related quantitative immunoelectrophoretic techniques have been used to elucidate the antigenic complexity of a reference preparation of capsular extract, potassium thiocyanate extract, lipopolysaccharide, heat-stable antigens, and free endotoxin from Pasteurella multocida serotype 1. The reactions of these cellular fractions, in crossed immunoelectrophoresis, with reference anti-whole cell immunoglobulins disclosed five antigens in the capsular extract, seven in the potassium thiocyanate extract, one to three in the lipopolysaccharide, three in the heat-stable antigens, and five in the free endoxin. Comparison of these reference antigen–antibody systems, in crossed immunoelectrophoresis, with intermediate gel containing either a reference anti-cell envelope or anticytoplasmic immunoglobulins not only revealed the presence of additional antigens but also gave insight into the probable cellular origins (i.e., cell surface, cell envelope, or cytoplasm) of various antigens unveiled by reference anti-whole cell immunoglobulins. Using the principle of tandem crossed immunoelectrophoresis and crossed-line immunoelectrophoresis the immunochemical relationships between the antigenic components of these reference antigen–antibody systems were established.

Immunogenicity of potassium thiocyanate extracted and electrofocused Pasteurella multocida X-73I antigens in chickens and mice

1982 ◽  
Vol 28 (11) ◽  
pp. 1219-1225 ◽  
Author(s):  
Kevin L. McKinney ◽  
Paul A. Rebers ◽  
Richard B. Rimler
Keyword(s):  
Antibody Response ◽  
Pasteurella Multocida ◽  
Potassium Thiocyanate ◽  
Particulate Fraction ◽  
Supernatant Fraction ◽  
Chicken Embryos ◽  
Isoelectric Points ◽  
Serotype 1 ◽  
Soluble Supernatant

The immunogenicity of antigenic fractions obtained by the extraction of Pasteurella multocida strain X-73 (serotype 1) with potassium thiocyanate (KSCN) was determined in chickens and mice. The initial KSCN extract was centrifuged at 105 000 × g, and the antigens were separated into a particulate fraction (40p) and a soluble supernatant fraction (40s). The ultracentrifuged fractions were further resolved by preparative electrofocusing. The 40p fraction was resolved into two subgroups having isoelectric points of 3.5–3.9 and 5.5–6.0; the 40s fraction was resolved into five subgroups ranging in isoelectric points from 4.4 to 9.0. The 40p fractions were antigenically similar and contained lipopolysaccharide (LPS) and protein. The 40s fractions were antigenically distinct from the 40p fractions and from each other; they contained proteins and polysaccharides but no LPS. The 40p antigens were strongly immunogenic in mice and chickens, whereas the 40s antigens were weakly immunogenic in chickens and not immunogenic in mice. The incorporation of Freund's complete adjuvant increased the immunogenicity of the 40s antigens in chickens. The 40p antigens induced greater frequencies of serological responses in chickens than the 40s antigens as detected by counterimmunoelectrophoresis and immunodiffusion. This suggested that the increased protection associated with the 40p antigens may have been the result of better antibody response. The toxicity of all the fractions was evaluated by determination of lethality for 10-day-old chicken embryos because of the sensitivity and reliability of the test. The 40p fraction had an LD50 = 0.38 μg, and the 40s fraction had an LD50 = 2.5 μg. Since the 40s fraction contained no detectable LPS, it is likely that two toxins are present, one which contains LPS and one which does not.

scholarly journals Induction of Protective Immunity in Rabbits by Coadministration of Inactivated Pasteurella multocida Toxin and Potassium Thiocyanate Extract

1998 ◽  
Vol 66 (8) ◽  
pp. 3788-3795 ◽  
Author(s):  
L. Z. Jarvinen ◽  
H. Hogenesch ◽  
M. A. Suckow ◽  
T. L. Bowersock
Keyword(s):  
Pasteurella Multocida ◽  
Systemic Disease ◽  
Potassium Thiocyanate ◽  
High Serum ◽  
Severe Liver ◽  
Igg Antibody ◽  
Local Immunity ◽  
Effective Response ◽  
Antibody Titers ◽  
Lung Infections

ABSTRACT Pasteurella multocida is a bacterial pathogen that causes rhinitis (snuffles), pneumonia, otitis media, septicemia, metritis, and death in domestic rabbits. Currently, there are no effective vaccines to prevent infection by this organism. Subcutaneous (s.c.) immunization with either exotoxin or thiocyanate extracts ofP. multocida induces partial protection in rabbits. Since disease begins at mucosal sites, induction of local immunity may be important in preventing systemic disease. Little is known concerning the efficacy of intranasal (i.n.) administration of these antigens in inducing protective mucosal immunity to P. multocida in rabbits. The purpose of this study was twofold: (i) to investigate the effectiveness of vaccination with purified P. multocidatoxin (PMT) and a potassium thiocyanate extract of P. multocida (CN) in combination and (ii) to evaluate the efficacy of administration of these antigens i.n. versus s.c. Forty-eight rabbits were randomly divided into eight different treatment groups. Rabbits received either one or both antigens by either s.c. or i.n. administration. Following vaccination, each group received an i.n. challenge of P. multocida. Rabbits vaccinated with both antigens i.n. or s.c. had a 100% survival rate, few or no bacteria in the liver and lungs, high serum immunoglobulin G (IgG) and IgM antibody titers, and significant numbers of IgG antibody-secreting cells (ASC) in the spleen and tracheobronchial lymph node. Rabbits vaccinated i.n. had significant nasal and bronchoalveolar lavage IgA antibody levels. Rabbits vaccinated with only one antigen, either PMT or CN, had lower antibody titers, moderate to severe liver and lung infections, and fewer ASC compared to rabbits receiving both antigens. Rabbits in the control groups had moderate to severe liver and lung infections. This study indicates that i.n. immunization with both PMT and CN induces an effective response against homologous P. multocidachallenge.

Purification of Pasteurella multocida antigens by ultracentrifugation and isoelectrofocusing

1982 ◽  
Vol 28 (5) ◽  
pp. 511-521 ◽  
Author(s):  
Kevin L. McKinney ◽  
Paul A. Rebers
Keyword(s):  
Crude Extract ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Pasteurella Multocida ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Potassium Thiocyanate ◽  
Polyacrylamide Gel ◽  
Specific Antigens ◽  
Ph Range

A procedure was developed to purify Pasteurella multocida X-73I antigens extracted by potassium thiocyanate. The crude extract was centrifuged at 105 000 × g; the antigens were then separated into a particulate (40p) fraction and a soluble (40s) fraction consisting of proteins and polysaccharides. These fractions were antigenically different.The ultracentrifuged antigens were resolved further by preparative isoelectrofocusing. The 40p antigens focused in a pH range of 3.0 to 6.0; distinctive proteins focused at pH's of 3.5, 3.6, and 3.8. The electrofocused 40p antigens were antigenically similar. The 40s antigens were initially electrofocused in a broad pH range and were found within a pH range of 4.4 to 9.0. The process was repeated with a narrower pH range and antigens that were focused in a narrower pH range could be separated and unique antigenic activities identified. Specific antigens from defined pH ranges were pooled and examined further by immunoelectrophoresis, analytical electrofocusing, and sodium dodecyl sulphate – polyacrylamide gel electrophoresis.

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