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Vaccination studies against experimental bovine Pasteurella pneumonia.

Vaccination-challenge experiments were conducted in colostrum-deprived calves to evaluate the efficacy of Pasteurella bacterins and vaccines against experimental pneumonic pasteurellosis. Calves were vaccinated with formalin-killed bacterins and live vaccines, then challenge exposed intratracheally with P. haemolytica or P. multocida. Infectious bovine rhinotracheitis virus was inoculated intranasally three to four days prior to P. haemolytica challenge-exposure. All calves were examined for macroscopic and microscopic lesions after being found dead or following euthanasia four to seven days after challenge exposure with the bacterial pathogen. Clinical, hematological, and pathological responses to challenge exposure in aluminum hydroxide absorbed P. haemolytica and P. multocida bacterin-treated calves were consistent with the pneumonic lesions of pulmonary pasteurellosis in the control calves. An oil-adjuvanted P. haemolytica bacterin limited clinical and pathological responses in the affected calves whereas a P. multocida oil-adjuvanted bacterin did not. Both clinical and pathological responses to challenge exposure in calves vaccinated with live Pasteurella vaccines were less severe than those of the control calves. Vaccine effectiveness appeared to be dose dependent.

Animals↗

A live Pasteurella haemolytica vaccine efficacy trial.

A live Pasteurella haemolytica serotype 1 vaccine was used in an efficacy trial conducted on 100 lightweight feeder calves purchased from a Florida ranch. Forty-one calves were inoculated with the vaccine intradermally in the neck. Fifty-nine calves served as nonvaccinated controls. Fourteen days later, the calves were shipped to an order buyer in eastern Tennessee, where the calves were mixed with 60 local calves in a community sale barn for 72 hours. After 3 additional days, the calves were shipped to a research feedlot in Bushland, Tex. They remained in the feedlot for 56 days, and the test was concluded 76 days after vaccination. The P haemolytica vaccine had no significant effect on performance, morbidity, or mortality. There was no significant difference between the vaccinated and nonvaccinated calves in the number of times Pasteurella was isolated. The calves became seropositive to bovine viral diarrhea virus, respiratory syncytial virus, and infectious bovine rhinotracheitis (IBR) virus during the 76-day experiment. All calves initially were seropositive to parainfluenza-3 virus. A virulent outbreak of IBR occurred 30 days after the calves arrived at the feedlot. Before the onset of IBR, the isolation of P haemolytica serotype 1 from nasal turbinates was rare (2 of 500 nasal swabs). After the IBR outbreak, P haemolytica serotype 1 was isolated from 40 of 92 calves.

Animals↗

An enzyme-linked immunosorbent assay to detect serum IgG to Pasteurella multocida in naturally and experimentally infected rabbits.

An enzyme-linked immunosorbent assay (ELISA) was evaluated for efficacy in detecting serum IgG against Pasteurella multocida in both naturally and experimentally infected rabbits. Blood samples and nasal cultures were taken concurrently from 58 rabbits from four conventional rabbitries. Nine rabbits from a pasteurella-free colony served as negative controls. Fifty-six rabbits were ELISA positive. Of these, 46 were P. multocida culture positive, 10 were culture negative. Two rabbits were ELISA negative, culture negative. There were no ELISA negative, culture positive animals. Serotyping by the gel diffusion precipitin test demonstrated that of the 44 typed P. multocida isolates, 57% were serotype 4, 27% were serotype 12 and 16% were serotype 3. In rabbits experimentally infected intranasally with P. multocida, serum IgG against P. multocida began to rise 21 to 33 days after infection and remained elevated until the animals were euthanized 90 days post infection. Two enzyme-linked immunosorbent assays were compared which used potassium thiocyanate extracts of different serotypes of P. multocida as antigen. The results obtained were similar, suggesting the presence of antigens common to both serotypes.

Animals↗

[Local and systemic immunization against experimental infection of mice with Pasteurella multocida].

Model experiments with mice are presented in which the defense against challenge infections has been checked after immunization with various living and killed vaccines of Pasteurella. A temperature sensitive Pasteurella mutant has proved to be superior to inactivated bacteria. It has been established that different routes of administration result in different effects. In this context, intraperitoneal, intramuscular and intratracheal administration have resulted in the highest protection rates, even when combined with oral and aerogeneous immunizations, which are of little efficacy when used exclusively. The importance of a change in the route of administration during repeated immunizations is discussed.

Administration, Intranasal↗

Experimental pneumonia in rabbits inoculated with strains of Pasteurella multocida.

Domestic rabbits were inoculated with either a 3:A or 3:D serotype of Pasteurella multocida by aerosol, intravenous, or intratracheal inoculation. Different colony forming units of P. multocida were used. Animals which died or were killed after the 14 day observation period were examined macroscopically and microscopically for lesions in the lower respiratory tract. Pneumonic lesions were most consistently produced in rabbits inoculated intratracheally with serotype 3:A. Pulmonary and pleural lesions were observed in some animals inoculated intravenously with serotype 3:A. Lesions were minimal in rabbits inoculated with serotype 3:D. Of the three routes of inoculation evaluated, the intratracheal route appeared to be the best method to produce Pasteurella-associated lesions in the lower respiratory tract.

Animals↗

[Antigenic structure of strains of Pasteurella multocida].

Studied was strain Pasteurella multocida-547 isolated from pigs affected with an acute Pasteurella infection. Serologically and immunologically its antigen was most closely related to the antigen of strain R-473. The main somatic antigen was 0:6, and the antigenic formula was 6:B. Homologous sera were used to demonstrate the presence of subsidiary antigens by which the newly isolated strain differed from strains R-473 (6:B), 1235 (6:E), and 989 (11:B). The immunodiffusion test with proper sera and heat-resistant water-soluble antigens could yield results that correlate with the typing of P. multocida strains by capsule and somatic antigens as established through other methods.

Acute Disease↗

Immunologic response to Pasteurella haemolytica and resistance against experimental bovine pneumonic pasteurellosis, induced by bacterins in oil adjuvants.

Immunogenicity of and protection afforded by Pasteurella haemolytica bacterins were studied in calves. Bacterins contained an aluminum hydroxide in gel (ALH) adjuvant or one of the following oil-in-water adjuvants: Freund's complete adjuvant (FCA), Freund's incomplete adjuvant (FIA), and trehalose dimycolate (TDM). On days 0 and 7, calves were vaccinated with phosphate-buffered saline solution (PBSS), a bacterin, or live P haemolytica. Transthoracic intrapulmonic challenge exposure was done on day 21. In 3 experiments, there were no significant (P greater than 0.05) differences between lung lesions induced in PBSS-or ALH bacterin-vaccinated calves. Both FCA and FIA bacterins significantly (P less than 0.05) enhanced resistance against challenge exposure. Resistance induced by FCA and FIA bacterins was comparable with that induced by vaccination with live P haemolytica. Calves vaccinated with FIA bacterin and challenge-exposed to P haemolytica at a concentration of 4.5 X 10(9) colony-forming units (4.5 times greater than used in the first 3 experiments) resisted challenge exposure similar to calves given live organisms. The TDM bacterin failed to enhance resistance. All bacterins caused a significant increase (P less than 0.05) in serum antibody to P haemolytica somatic antigens, as measured by a quantitative fluorometric immunoassay. Pasteurella haemolytica leukotoxin neutralizing antibody titers did not increase significantly (P greater than 0.05) in sera after vaccination with any bacterin. Vaccination with FCA and FIA bacterins resulted in a significant increase (P less than 0.001) in serum antibody to a carbohydrate-protein subunit of P haemolytica, as measured by an enzyme-linked immunosorbent assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗

Laboratory and field trials with formalin-inactivated Escherichia coli (O78)-Pasteurella anatipestifer bacterin in white pekin ducks.

A combination Escherichia coli serotype O78 and Pasteurella anatipestifer bacterin was developed and tested in white pekin ducks in laboratory and field trials. Inoculations with bacterin at 2 and 3 weeks of age provided significant protection against challenge with virulent E. coli O78 and Pasteurella anatipestifer serotypes 1, 2, and 5. No significant cross-protection was observed against heterologous E. coli serotypes, although there was a slight reduction in mortality in ducklings challenged with E. coli serotypes O2a and O119. In field trials, the E. coli-P. anatipestifer bacterin produced significant reduction of mortality in commercial white pekin ducks compared with P. anatipestifer bacterin.

Animals↗

Non-specific resistance of mice to Pasteurella haemolytica.

A technique for challenge infection of mice with Pasteurella haemolytica is described. Mice were challenged intraperitoneally with P haemolytica in gastric mucin, and viable counts were performed 6 h later on liver suspensions. Viable counts of P haemolytica in the livers of unvaccinated control mice had increased 6 h after infection. Mice were injected subcutaneously with a commercial pasteurella vaccine at various time intervals before challenge. In those given vaccine two weeks, one week, 48 h and 1 h before challenge, viable counts of P haemolytica in the liver increased as for control mice. However, if the time interval between administration of vaccine and challenge was 12 or 24h, viable counts of P haemolytica in the liver decreased after challenge. Subcutaneous inoculation with the nonbacterial components of the vaccine 24 h before challenge did not cause such an effect. Inobulation with the vaccine 24h before challenge with Escherichia coli resulted in an increase in the LD50 for that organism, indicating that the effect is probably non-specific.

Animals↗

Pasteurella multocida infection in the domestic rabbit: immunization with a streptomycin-dependent mutant.

Fourteen Pasteurella multocida-free rabbits were inoculated intranasally with a streptomycin-dependent mutant of P. multocida serotype 12:A. Vaccinations with approximately 10(8) colony forming units were done on days 0, 14 and 28. Two weeks later the animals were separated into groups, which included 12 rabbits divided into two control groups of six unvaccinated Pasteurella-free animals. Seven vaccinated rabbits were challenged intranasally with the homologous virulent parent strain and the other seven vaccinates were challenged with a virulent strain of serotype 3:A. Rabbits were necropsied two weeks later. The vaccinated group challenged with the parent strain showed a more rapid nasal clearance of the organism than the vaccinated group challenged with the heterologous strain. However, the number of positive cultures of P. multocida recovered from tissues post-challenge were similar in vaccinated and control animals. In a significant number of animals, vaccination with serotype 12:A induced detectable antibody production to somatic antigens of both 12:A and heterologous strain 3:A.

Animals↗

Calfhood immunization against an experimental Parainfluenza and Pasteurella challenge.

An inactivated vaccine including Parainfluenza-3, PASTEURELLA MULTOCIDA and PASTEURELLA HEMOLYTICA stimulated good immunity in calves against an experimental shipping fever challenge. Calves were first vaccinated when 4 to 10 weeks old and revaccinated at weaning age. There were higher HI titers, lower temperatures, and less lung lesions recorded in the vaccinated group when compared to the control animals.

Animals↗

In vitro killing of Pasteurella multocida: the effect of rabbit granulocyte and specific antibody source.

In vitro granulocyte-killing assays were performed to examine the ability of granulocytes from pasteurella-free or immunized rabbits, the in combination with specific immune serum, to kill Pasteurella multocida. Granulocytes from healthy rabbits and from rabbits with P multocida infections were equally competent. Granulocyte source, serum source, and specific antibody titer had no effect on granulocyte phagocytic activity. Moreover, serum containing specific antibody and complement supported the growth of the bacterium. These data suggest that chronic P multocida infections are not attributable to defective granulocytes or lack of serum antibody production.

Animals↗

Bovine herpesvirus-1 vaccination against experimental bovine herpesvirus-1 and Pasteurella haemolytica respiratory tract infection: onset of protection.

The onset of protection offered by intranasal vaccination with attenuated bovine herpesvirus-1 (BHV-1) was studied in 18 calves given a virulent BHV-1 aerosol challenge inoculum and an aerosol challenge exposure to Pasteurella haemolytica. Calves challenge exposed with virus 3, 7, 11, 15, or 19 days after vaccination and challenge exposed 4 days later with Pasteurella haemolytica did not develop viral-bacterial pneumonia, whereas 2 of 3 control calves died of fibrinous bronchopneumonia 40 and 60 hours after the bacterial aerosol and the 3rd control calf had similar lesions. All vaccinated and control calves had detectable amounts of interferon at the time of viral challenge exposure. Protection was observed before detection of neutralizing antibodies to BHV-1 in nasal secretions or in serum. Protection was therefore present from day 3 through day 19 after vaccination, but the mechanism could not be explained completely by neutralizing antibody or interferon.

Animals↗

The usefulness of the API 20 E classification system in the identification of Actinobacillus actinomycetem comitans, Actinobacillus seminis and Pasteurella haemolytica.

The prepuces of lambs aged 6--8 months and semen of 2 adult rams were found to be infected with gram negative, non-motile, non-haemolytic, pleomorphic bacilli. These organisms were compared with those of known strains of actinobacillus actinomycetem comitans. Actinobacillus seminis and Pasteurella haemolytica, using the API 20 E classification system. Applying the principles of numerical taxonomy, the majority of suspected strains of A. seminis could be classified as A. actinomycetem comitans and 3 examples as Histophilus ovis. Although some of the suspected strains of A. seminis could be classified as P. haemolytica, obvious differences between the genera Actinobacillus and Pasteurella were evident.

Actinobacillus↗

Bovine pneumonic pasteurellosis: effect of vaccination with live Pasteurella species.

Experimental bovine pneumonic pasteurellosis was induced in beef calves by a transthoracic challenge exposure with Pasteurella haemolytica serotype 1 or P multocida type 3. Challenge exposure lesions were quantified by a lesion scoring system based on size and extension of lesions with larger scores assigned to the more severe lesions. Calves inoculated with live Pasteurella sp by aerosol or parenteral routes developed high serum antibody titers to the homologous organism, as determined by a quantitative fluorometric procedure. Mean lesion scores were approximately 2 to 20 times higher in control than those in vaccinated calves. There was a significant correlation (P less than 0.05) between high serum antibody titers at the time of challenge exposure and a low lesion score in 4 of 6 experiments.

Aerosols↗

Response of young chickens and turkeys to virulent and avirulent Pasteurella multocida administered by various routes.

Groups of 20 young chickens and turkeys obtained from commercial flocks and placed in pens with pine shavings over concrete floors or from the hatchery and grown on wire, were exposed to avirulent Pasteurella multocida serotype 3,4 cross or virulent P. multocida serotype 3 by drinking water, ocular, subcutaneous, intracutaneous, or palatine cleft routes. A secondary exposure to virulent P. multocida serotype 1 by the palatine cleft route was given 14 days later, except that birds exposed by palatine cleft on the initial exposure were reexposed by drinking water. There was a group of each species for each floor type and each exposure route except palatine cleft, which was performed on litter only. Chickens were more resistant to initial exposure to virulent P. multocida by all routes than turkeys. Chickens showed much less resistance to subsequent exposure to P. multocida serotype 1 than turkeys after exposure to the avirulent organism by the drinking-water route. Avirulent P. multocida administered to chickens or turkeys by subcutaneous or intracutaneous routes and turkeys by drinking water route produced a high degree of resistance to subsequent exposure to P. multocida serotype 1. In a second experiment it was found that wire floors caused many more breast blisters than pine shavings on Pasteurella-exposed or unexposed chickens.

Animals↗

Isolation of toxigenic strains of Pasteurella multocida from lungs of pneumonic swine.

Lungs from 113 pneumonic pigs were examined for Pasteurella multocida. The lungs were smeared directly onto blood agar and homogenized in brain-heart infusion broth and then inoculated intraperitoneally in mice. Pasteurella multocida isolates were typed for serotypes A (by hyaluronidase inhibition of capsule) and D (by acriflavine autoagglutination). Strains were tested for toxin production by intradermal injection of 0.2 ml of filtered 24-hour culture supernatants into guinea pigs. Most lungs (70.8%) yielded isolations. Most isolants (87.5%) were type A and 12.5% were type D. Of the type D strains, 80% were toxigenic. Of the type A isolants, 18.2% were toxigenic.

Animals↗

Comparison of enzyme-linked immunosorbent assay and indirect hemagglutination test for quantitating antibody responses in chickens against Pasteurella multocida.

An indirect enzyme-linked immunosorbent assay (ELISA) was developed to measure humoral antibody responses of chickens against Pasteurella multocida. A standard indirect hemagglutination (IHA) test was used to compare serologic results with those of ELISA. The ELISA was also used following challenge with P. multocida to compare the efficacy of three commercial fowl cholera vaccination regimens. Although antibody titers measured by ELISA and IHA were highly correlated, ELISA was at least twice as sensitive as IHA. Antibody measured by ELISA and IHA also correlated significantly with protection against P. multocida challenge. No mortality occurred in any of the three vaccinated challenged groups. However, control unvaccinated chickens experimentally infected with P. multocida developed signs of acute pasteurellosis and died by the 10th day post-challenge. Impression smears made of hepatic tissue from all chickens were stained (Wright's stain), and typical bipolar rods characteristic of Pasteurella were identified in smears from unvaccinated challenged controls only.

Animals↗