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At least 649 records · Page 36Linked to original sources
PubMed1994

Use of a 35.5 kDa cell membrane composition of Pasteurella multocida and an anti-idiotype antibody to induce protective immunity in leghorn chickens.

Using polyacrylamide gel electrophoresis and Western blot analysis, a 35.5 kDa cell membrane composition of Pasteurella multocida ATCC 11039 was identified as a dominant epitope of the bacterium. Mice inoculated with inactivated whole bacteria produce antisera primary reactive with the 35.5 kDa component (Pm35.5) in Western blot analysis. The outer membrane component was composed primarily of protein but did have lipopolysaccharide present at 133 micrograms mg-1 protein. In challenge trials (Trial 2), groups of white leghorn chickens vaccinated with Pm35.5 or an anti-idiotypic antibody (using Pm35.5 as the original antigen) had significant difference in mortality of 3.2% and 48.3%, respectively, when compared with unvaccinated controls (99%). Mortality in a group of chickens receiving a commercially available bacterin (9.2%) was higher but not significantly different from the mortality of the Pm35.5 vaccinated group (3.2%).

Animals↗
PubMed1994

Immune responses of piglets to Pasteurella multocida toxin and toxoid.

Experimental atrophic rhinitis (AR), serum antibody titres and in vitro lymphoproliferation to Pasteurella multocida derived toxin (Pm-T) were studied in piglets. Specific immune responses to Pm-T and Pm-T induced conchae atrophy were compared with AR immunity. This immunity was initiated by the Nobi-VAC AR-T vaccine administered at various times with respect to Pm-T challenge. Animals challenged with Pm-T developed conchae atrophy, but no antibodies nor cellular immune responses to Pm-T were detected. Vaccination 3 weeks before Pm-T challenge protected pigs against breakdown of nasal bony tissues. This protection was accompanied by an increase of serum antibodies and in vitro lymphoproliferation to Pm-T. Animals vaccinated 10 days before or after Pm-T challenge also had antibodies and cellular immune responses. However, these animals developed AR. In vitro, Pm-T appeared mitogenic for quiescent (non-immune) peripheral lymphocytes and Concanavalin A stimulated lymphocytes from some pigs. These in vitro lymphoproliferative responses could be partly abrogated by the addition of monomorphic anti-swine major histocompatibility complex class II DQ and DR specific monoclonal antibodies. We conclude that Pm-T is poorly immunogenic in vivo and does not initiate a protective Pm-T specific immune response. Pigs were protected from AR by vaccination, but protection was dependent on the timing of vaccine administration. We speculate that Pm-T modifies the immune response such that the response is not directed towards the toxin but to an unidentified component in the nose of piglets.

Animals↗
PubMed1995

Occurrence of T lymphocytes in perivascular regions of the lung after intratracheal infection of swine with Pasteurella multocida.

The defence mechanisms of the lung acting against airborne antigens (bacteria) are not yet fully understood. Therefore, we investigated cell-derived immunological processes in porcine pulmonary tissue. The incidence and distribution of leukocyte subpopulations in lungs, lung lymph nodes and tonsils from eight animals intratracheally infected with Pasteurella multocida (P.m.) type A wild strain and from four non-infected control animals were established by immunohistochemistry, using a panel of monoclonal antibodies (mAbs) against specific porcine leukocyte antigens: MHC Class II (MHC II), SWC1, SWC3a, CD2a, CD4a, CD8b. In the test animals, a broad layer of SWC1+ cells and SWC3a+ cells emerged in the subsinusoidal region of lung lymph nodes and in the bronchoalveolar spaces of the lung as early as 4 and 24 h after infection, and in the subsinusoidal regions of the lung lymph nodes only very few cells could be stained with the mAb against the MHC II antigen. We could not detect any MHC Class II+ cells in the bronchoalveolar spaces at this time of the investigation. In the course of the disappearance of the SWC1+ cells and SWC3+ cells, CD2a+, CD4a+ and, in lower numbers, CD8b+ T lymphocytes seemed to concentrate in the perivascular and, partially, the peribronchial regions of the lung 72 h after infection. By means of the double immunostaining method, we could demonstrate an accumulation of CD4a+ cells and CD8b+ cells after infection which lacked the SWC1 antigen, indicating that these cells were activated T lymphocytes. The same cell types (SWC1-/CD4a+ and SWC1-/CD8b+ cells) as well as CD8b-/CD4a+ cells could be observed in the interstitium of the lung 72 h after infection.

Animals↗
PubMed1996

Effect of Pasteurella multocida vaccination on buffalo polymorphonuclear hydrogen peroxide and nitric oxide production.

An attempt was made to investigate the effect of Pasteurella multocida on certain microbicidal reactive oxygen and nitrogen intermediates released by the polymorphonuclear cells (PMNs) from the vaccinated animals. The PMNs from the peripheral blood of both control and experimental buffaloes vaccinated against haemorrhagic septicaemia were isolated. PMNs from control animals upon activation with P. multocida lipopolysaccharide (LPS) and live P. multocida cells generated higher levels of hydrogen peroxide (H2O2) and nitric oxide (NO-) than the non-activated cells (P < 0.01). In the presence of P. multocida LPS, PMNs from animals vaccinated against haemorrhagic septicaemia generated significantly higher H2O2 (P < 0.05) and NO- (P < 0.01) than the PMNs from control animals. L-Arginine when added to the activation medium enhanced the production of NO- in a dose-dependent manner. This indicated the role of arginine in NO- production. The study suggested that buffalo PMNs possessed a potent oxidant defence system even in the presence of P. multocida, an antiphagocytic bacterium.

Animals↗
PubMed1996

Immunity and vaccine development in Pasteurella multocida infections.

The role of LPS in immunity was studied using monoclonal antibodies (MAbs) and active immunisation experiments. A panel of six MAbs produced against Pasteurella multocida serotype B:2 reacted with the LPS of serotypes B:2 and B:5, but not with other serotypes. The MAbs could opsonise P. multocida for phagocytosis by mouse macrophages, but were not bactericidal in the presence of complement. They conferred only partial passive protection in mice. Similar results showing only partial protection were obtained when purified LPS was used to actively immunise mice prior to challenge, suggesting that LPS plays a partial role in immunity to infection. The aroA gene from P. multocida serotypes A:1 and A:3 was cloned and inactivated by insertion of a kanamycin resistance gene. The mutated gene was re-introduced onto the chromosome by allelic exchange. The resultant aroA mutants were highly attenuated in a mouse model system, with a 6-log decrease in ID50. Virulence could be restored by complementation with a functional aroA gene. Mice immunised with two doses of the live mutants were protected against lethal challenge with the homologous parental strain, but not against the heterologous strain. P. multocida A:1 and A:3 expressed unique proteins when grown in iron-restricted medium. Moreover, the outer membrane (OM) fractions of these cells contained novel proteins of 75 kDa, 85 kDa and 94 kDa molecular mass. Mice were immunised with OM fractions prepared from serotype A:3 grown in iron-restricted (OM Fe-) or iron-replete (OM Fe+) media. When low challenge doses were used, both immunogens protected mice against serotype A:3, but only the OM Fe- fraction protected mice against heterologous challenge with serotype A:1. When higher challenge doses were used, only partial protection was observed.

3-Phosphoshikimate 1-Carboxyvinyltransferase↗
PubMed1987

Failure of oil adjuvants to enhance immunity induced in mice by an inactivated rabbit Pasteurella multocida vaccine.

Mice were immunized with different preparations of a killed whole cell Pasteurella multocida vaccine and challenged with successive tenfold dilutions of the homologous strain. Vaccines adjuvanted with mineral oil did not confer better protection against death than non-adjuvanted suspensions, even when the mice were challenged at 3 months after vaccination. Very good protection was obtained when the non-adjuvanted suspension had been administered twice.

Adjuvants, Immunologic↗
PubMed1990

Protective effect of inactivated Pasteurella haemolytica bacterin challenged in bovine herpesvirus-1 experimentally infected calves.

Formalin-inactivated and sonicated Pasteurella haemolytica bacterins were prepared from 1 h cultures of bacterial pellets in RPMI-1640 medium containing 7% fetal calf serum. The bacterial pellets were obtained from logarithmic phase growth of the organism by centrifugation. The protective effect of the vaccine was evaluated in 43 specific-pathogen-free Hereford crossbred calves and yearlings in three experiments. Cattle were either single vaccinated or boosted via three routes; intratracheally (i.t.), intranasally (i.n.) or intramuscularly (i.m.), using low or high doses. The two low-dose groups were also given supernatant by the same routes and volume as the bacterin. Cattle were challenged by P. haemolytica in aerosol at 24 or 39 days after last vaccination. To enhance the susceptibility of the cattle to this challenge, the cattle were exposed to bovine herpesvirus-1 aerosol 4 days before the bacterial challenge. The extent of pneumonia was significantly less in three groups of cattle (i.n.-i.n.,i.m.-i.n.,i.m.-i.m.) boosted with high dose of the bacterin than in the controls. Protection was observed when challenge isolates were heterologous or homologous to the isolates used to prepare the bacterins. It was also observed that the level of complement fixing antibody or anticytotoxin activity to P. haemolytica did not correlate with the degree of protection.

Administration, Intranasal↗
PubMed1991

Vaccine containing iron-regulated proteins of Pasteurella haemolytica A2 enhances protection against experimental pasteurellosis in lambs.

A vaccine containing sodium salicylate extract (SSE) of Pasteurella haemolytica A2 cells grown in a medium chemically depleted of available iron by the addition of alpha alpha dipyridyl to induce iron-regulated proteins (IRPs) conferred protection to specific pathogen-free (SPF) lambs exposed to an aerosol of P. haemolytica A2. The disease score in these lambs was significantly lower (p less than 0.005) than those in unvaccinated lambs or in lambs immunized with SSE prepared from cells grown in iron-replete medium. Immunoblotting of sera from these SPF lambs against whole cell antigens of P. haemolytica A2 grown under iron-restricted conditions demonstrated that antibodies to IRPs were present only in the sera of animals immunized with SSE-IRP. The antibody profile of sera from the SSE-IRP group was similar to that obtained with serum from a lamb which had recovered from P. haemolytica A2 disease produced experimentally. Negligible levels of cytotoxin-neutralizing and bactericidal antibodies were detectable in the SSE-IRP group and therefore appear not to be involved in the protection observed in this experiment.

Animals↗
PubMed1995

Pili of Pasteurella multocida of porcine origin.

Using electron microscopy, pili with at least two distinct morphologies were observed on strains of Pasteurella multocida isolated from pigs with atrophic rhinitis. Rigid pili were found on 60-80% of all cells observed. These pili had a strong tendency to lie flat along the side of the outer cell membrane of P. multocida and as a result frequently were difficult to see. After growth in vitro, piliated P. multocida cells produced few pili (approx. 3-5 per cell). Heavily piliated cells were occasionally observed. The second type of pili were curly and also were difficult to visualize. Cells from cultures containing piliated cells failed to attach to red blood cells and to immobilized mucus.

Animals↗
PubMed1984

Experimental infection of sheep with Mycoplasma ovipneumoniae and Pasteurella haemolytica.

A group of Caesarian-derived, colostrum-deprived lambs was inoculated intranasally and intratracheally with a virulent Mycoplasma ovipneumoniae isolate selected from ovine mammary studies and propagated in an ovine mammary gland. Other groups of lambs were inoculated with M. ovipneumoniae in combination with Pasteurella haemolytica type Al or P. haemolytica alone. The M. ovipneumoniae isolate alone did not induce any specific pneumonic lesions in the lambs and when combined with P. haemolytica type Al did not increase the severity of the P. haemolytica-type lesions. Fifty percent of lambs inoculated with P. haemolytica developed a purulent and exudative bronchopneumonia with pleurisy and high titres of P. haemolytica were recovered from these lesions.

Animals↗
PubMed1984

A protective antigen for turkeys purified froma type 1 strain of Pasteurella multocida.

A protective antigen was purified from a saline extract of a Type 1 strain of Pasteurella multocida by chromatographic methods, and its chemical and immunological characteristics were studied. Three proteins peaks were obtained from crude extract by gel filtration with Sephadex G-200. A bacteria-specific antigen was detected only in the first peak fraction, which, after passing through an immunoabsorbent column to remove any components originating from the growth medium, was absorbed onto DEAE-cellulose followed by elution with a gradient of NaCl. From the first peak fraction of the gel filtration, 4 protein peaks were obtained, the second and third peaks being the major ones. Carbohydrate/protein ratios of the peak fractions varied from 0.06 to 1.0. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that 2 proteins of molecular weights 44 000 and 25 000 were present in all the fractions. The 4 DEAE-cellulose fractions (DP-1 to DP-4) contained a single antigenically identical material, and induced protective immunity in turkeys against challenge exposure. The second peak fraction from DEAE-cellulose (DP-2) protected turkeys when subcutaneously injected as 2 doses of 10 micrograms protein with a 14-day interval between doses. The DP-2 fraction induced antibodies in rabbits which formed a single precipitin line against the crude extract. The purified antigen (DP-2) from a Type 1 strain was antigenically distinct from a similar antigen purified from a Type 3 strain; there was no significant cross protection in turkeys between the 2 antigens. These results indicate that protective antigens purified from soluble extracts of a Type 1 or Type 3 strain possess similar physicochemical properties, but that they are immunologically distinct from each other.

Animals↗
PubMed1985

Pasteurella haemolytica: purification of saline-extractable proteins by isoelectrofocusing.

A procedure was developed for separating antigens associated with a saline extract of Pasteurella haemolytica serotype 1. Seven antigens were identified by immunoelectrophoresis to be associated with the extract. The extract was subjected to preparative isoelectrofocusing in a pH range of 3-10. The majority of extracted proteins were found to have pI's of 4-6, whereas the carbohydrate antigen(s) were distributed over a pI range of 3.0-8.0. The fractions that were of interest were pooled and refocused in a narrower pH range to improve resolution of the protein antigens. Specific antigens from defined pH ranges were pooled to form 6 antigen groups. These antigen groups were examined further by immunoelectrophoresis, analytical isoelectrofocusing, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The molecular weights of the proteins found in the capsular extracts ranged from 33 k to greater than 80 k. Injection of mice with capsular extract or antigen Groups 1-6 in Freund's incomplete adjuvant resulted in a serum antibody response to the various antigens as detected by an enzyme-linked immunosorbent assay. Significant protection (P less than 0.05) against challenge with virulent P. haemolytica was seen in mice injected with antigen Groups 2 and 4. Six calves were immunized with saline extract. These calves had greater resistance to experimental pneumonic pasteurellosis than did 6 non-vaccinated calves. A serum antibody response to the crude extract and to each antigen group was detected in vaccinated calves by an enzyme-linked immunosorbant assay.

Animals↗
PubMed1986

Immunogenicity of potassium thiocyanate extract of type A Pasteurella multocida.

The immunogenicity and cross-protectivity of potassium thiocyanate (KSCN) extracts from Type A Pasteurella multocida strains were evaluated in mice. Mice administered KSCN extracts showed signs of depression for several hours following immunization. These extracts induced protection against challenge with the virulent homologous bacterium although many of the surviving mice were clinically ill up to 72 h after challenge exposure. There was no consistent reciprocal protection between different strains of the serotype indicating lack of correlation between serotype and cross-immunogenicity. Antigenic analysis of KSCN extracts by crossed immunoelectrophoresis techniques revealed greater than or equal to 25 different antigenic components. When the antigenic content of P-2383 and P-1062 KSCN extracts were compared, most antigenic components were common to both; however, two strain-specific and antigenic components were identified with homologous and heterologous antisera. One component present in each of the extracts gave a precipitation line similar to that produced by a crude lipopolysaccharide extracted from the organism. This component was isolated by sucrose density gradient centrifugation. The component isolated from P-2383 KSCN extract sedimented rapidly, indicating a high molecular weight material, and contained 12% carbohydrate and 27% protein. Immunization of mice with this component induced resistance to challenge with the homologous strain and some degree of protection against certain heterologous strains.

Animals↗
PubMed1986

The effect of parainfluenza virus type 3 and Pasteurella haemolytica on oxygen-dependent and oxygen-independent bactericidal mechanisms of ovine pulmonary phagocytic cells.

Groups of caesarean-derived, colostrum-deprived lambs were inoculated by the intratracheal route with Pasteurella haemolytica, either alone or 4 or 6 days after the inoculation of parainfluenza virus type 3 (PI3). Other groups were inoculated with PI3 followed by veal infusion broth, or with uninfected cell culture fluid followed by veal infusion broth (controls). All lambs were killed 24 h after the second inoculation. Pulmonary phagocytic cells were recovered by lavage and separated into alveolar macrophage (AM) and neutrophil fractions by density gradient centrifugation. Bacterial proliferation was detected in the lungs of all five lambs inoculated with P. haemolytica 6 days after PI3 but in only one of five inoculated with P. haemolytica 4 days after PI3 and one of five inoculated with P. haemolytica alone. The number of phagocytic cells recovered from the lungs was highest in animals inoculated with P. haemolytica 6 days after PI3 and, overall, a greater number of both AM and neutrophils was recovered from the lungs of animals where bacterial proliferation occurred (greater than 10(5.0) P. haemolytica 100 g-1 lung) than from those that controlled the bacterial infection. Oxygen-dependent bactericidal activity of AM and neutrophils was measured by chemiluminescence. Infection with PI3 and P. haemolytica increased the chemiluminescence responses. The highest responses were recorded from lambs inoculated with P. haemolytica 6 days after PI3, the group where pulmonary clearance was poorest. Overall, responses were higher in lambs in which bacterial proliferation occurred than in those that controlled the infection. On the other hand, oxygen-independent bactericidal activity, measured by the direct effects of neutrophil lysates on Escherichia coli, was lowest in lambs inoculated with P. haemolytica 6 days after PI3 and was lower in lambs where bacterial proliferation occurred.

Animals↗
PubMed1986

Measurement of Pasteurella haemolytica-specific lung and serum antibodies by ELISA.

The modified enzyme-linked immunosorbent assay (ELISA) was used to determine the relative quantities of class-specific antibodies to Pasteurella haemolytica. IgG1, IgG2 and IgA were present in significantly higher quantities in bronchoalveolar washings (BAW), but in decreasing quantities, respectively; IgM was present in very low amounts. IgM, IgG1 and IgG2 were present in serum, again in decreasing quantities, respectively. IgA antibody quantities were lowest in serum. The indirect antibody ELISA was found to be superior to the indirect bacterial agglutination (IBA) technique for determining antibody titres against P. haemolytica.

Agglutination Tests↗
PubMed1987

Bordetella bronchiseptica and toxigenic type D Pasteurella multocida as agents of severe atrophic rhinitis of swine.

Bordetella bronchiseptica and toxigenic type-D Pasteurella multocida were cultured from pigs in each of five herds diagnosed as having severe atrophic rhinitis (AR). B. bronchiseptica alone, P. multocida alone, or both organisms isolated from four herds were inoculated intranasally into 1-week-old gnotobiotic pigs which were necropsied 4 weeks post-inoculation (PI). Nasal turbinate atrophy in B. bronchiseptica-inoculated pigs was moderate to severe, while P. multocida-inoculated pigs had slight to severe atrophy. Pigs inoculated with both organisms had moderate to complete turbinate atrophy. P. multocida was reisolated at necropsy from all pigs receiving the organism except those having no turbinate damage. B. bronchiseptica and P. multocida from a fifth herd were simultaneously inoculated into six naturally farrowed 6-day-old SPF pigs. Necropsy performed 4 weeks PI revealed severe to complete turbinate atrophy. Nasal turbinates were normal for control pigs in both experiments.

Animals↗
PubMed1988

A rapid micro-method for the study of antibody-mediated killing of bacteria, with specific application to infection of sheep with Pasteurella haemolytica.

A micro-titration plate bactericidal assay was developed to measure complement-dependent antibody-mediated killing of Pasteurella haemolytica. Sera and lung washings from specific pathogen-free (SPF) lambs convalescent from a challenge with live, virulent P. haemolytica were bactericidal in the presence of complement. Similar samples from naive SPF lambs had no such activity. Purified IgG derived from a convalescent lamb serum was as bactericidal as the whole serum. Absorption of convalescent serum with lipopolysaccharide from P. haemolytica abolished bactericidal activity, suggesting that this antigen may be a target for antibody in the bactericidal complex.

Animals↗
PubMed1989

Comparing different isolates of Pasteurella haemolytica from beef calves using their in vitro antimicrobial sensitivity patterns.

In vitro studies, using disc diffusion and agar dilution techniques, were carried out to compare susceptibilities to selected antimicrobial agents of 30 isolates of Pasteurella haemolytica from healthy calves and 30 isolates from calves with transit fever. There was no difference in susceptibility patterns between isolates from healthy calves and isolates from diseased calves or between isolates of serotype A1 and isolates of serotype A2. Penicillin resistance was associated with production of beta-lactamase.

Animals↗