Virulence and lienotoxicity of Bordetella bronchiseptica in mice

Whole-cell suspensions (WCSs) and cell-free sonicated extracts (SEs) of seven Bordetella bronchiseptica strains were studied for lethality and lienotoxicity in mice. Lethality was assessed after intravenous and intracerebral inoculation, and lienotoxicity by splenic atrophy after intravenous inoculation. The strains represented phase I isolates with or without cytotoxin production, their phase III subcultures and a phase IV variant. The lethality and lienotoxicity of the SEs were in close positive correlation with cytotoxin production. The WCSs of all phase I strains were lethal, irrespective of their cytotoxin- and lienotoxin-producing ability. The only difference was that cytotoxic phase I strains caused splenic atrophy while the noncytotoxic phase I strain induced splenic hypertrophy in the surviving mice. The WCSs of phase III and IV variants were non-lethal and caused splenic hypertrophy even though all but one of them showed some cyto- and lienotoxic activity when their SEs were tested. The results indicate that B. bronchiseptica possesses two different mouse lethal factors: one seems to be identical with the cytotoxin, the other is associated with cell integrity and viability and, presumably, propagation in vivo. It also follows from the results that only the SEs are suitable for accurate determination of the lienotoxin-producing ability of B. bronchiseptica.     

Effects of vaccine route and dosage on protection from rabies after intracerebral challenge in mice

OBJECTIVE: To evaluate the effect of various routes of administration and number of doses of 3 commercially produced rabies vaccines on serum antibody responses and protection in mice challenged by intracerebral injection with fixed-strain rabies virus. ANIMALS: 2,213 mice. PROCEDURE: Inactivated, adjuvanted rabies vaccines were administered to mice in either 2, 1, or 0 (control) doses via IP, IM, and SC routes, and mice were challenged intracerebrally with fixed-strain rabies virus. RESULTS: Vaccination route and dose number significantly influenced serum antibody responses and protection from rabies virus challenge, independent of vaccine strain origin and mouse strain, although mouse age significantly affected results. Extended challenge studies revealed that IM vaccination of mice resulted in the highest serum neutralizing antibody responses and protection levels equivalent to IP vaccination. Even multiple doses administered SC (a vaccination route used in dogs) resulted in poor serum anti-rabies neutralizing antibody responses in mice and were far less protective than other routes. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggest possible improvements for the current rabies vaccine potency test in mice by using 1 dose, the IM route, and a delayed time of challenge. These modifications would more closely model vaccination practices in target species and yield more accurate information regarding primary immunogenicity of a vaccine.     

Intranasal infection with Bordetella bronchiseptica in gnotobiotic piglets.

Nineteen gnotobiotic piglets, four to six days old, from three different litters, were inoculated intranasally on three consecutive days with a pure culture of Bordetella bronchiseptica. Necropsy 10 to 30 days after inoculation revealed atrophic rhinitis lesions in all and pneumonia in 13 (73 per cent)infected piglets. One died of septicaemia two days after inoculation. Agglutinating antibodies were detected in the sera of all piglets necropsied from 17 to 30 days after inoculation. Five control piglets showed no lesions or serological changes.     

Experimental respiratory disease in dogs due to Bordetella bronchiseptica.

Young dogs of two age groups, six weeks and 12 weeks respectively, were infected by aerosol with a strain of Bordetella bronchiseptica which had been isolated from a dog with pneumonia. Clinical respiratory disease characterised by coughing and in some cases purulent nasal discharge was induced in both groups of infected dogs and also in dogs kept in contact. B bronchiseptica was recovered from the nasal cavity, trachea, bronchi and lung parenchyma of infected and contact animals. At necropsy, masses of Gram-negative bacteria were found trapped in the cilia of the respiratory epithelia and there was an exudate containing neutrophils in the mucosae of the respiratory tract at all levels. A close similarity was noted between the lesions produced in the dog and those described in pertussis infection in man. Experimental respiratory disease in the dog due to B bronchiseptica may offer a model system for the study of the human disease.     

Immunogenicity and safety of an attenuated Bordetella bronchiseptica vaccine in pigs

The immunogenicity and safety of an attenuated Bordetella bronchiseptica vaccine for swine atrophic rhinitis (AR) was evaluated in 22 hysterectomy-produced, colostrum-deprived pigs and 18 conventional pigs. None of 8 pigs inoculated at 7 days of age intranasally with greater than or equal to 3 X 10(5) colony-forming units (CFU) of vaccinal strain/pig and 2 of 5 pigs inoculated at 7 days of age intranasally with 3 X 10(4) CFU of the vaccinal strain/pig developed AR after intranasal challenge exposure with a virulent strain at postinoculation week (PIW) 3. The remaining 3 vaccinated pigs and 4 nonvaccinated pigs developed AR. Thirteen pigs were inoculated intranasally with 3 X 10(6) to 3 X 10(9) CFU of the vaccinal strain at 7 days of age. At PIW 12, the pigs were killed and necropsied. None of the pigs had clinical signs of AR and/or pneumonia. Virulence was studied by transmission of vaccinal strain through 3 serial growing passages on the nasal mucosa of a litter of hysterectomy-produced colostrum-deprived pigs. Inoculum (nasal swab samples from 2 pigs 4 days after inoculation with 10(8) CFU of vaccinal strain at 5 days of age) was inoculated into the nasal cavity of 2 nonvaccinated pigs. This procedure was repeated 3 times. After the 1st passage, the vaccinal strain was recovered on postinoculation day 4, but after postinoculation day 4, the vaccinal strain was not recovered until the end of the 3rd passage. Turbinate atrophy or pneumonia was not recognized in these inoculated pigs. The vaccinal strain provided immunogenicity without ill effects.     

Inhibitory effect of sulfamonomethoxine on capsule formation of Bordetella bronchiseptica.

Bordetella bronchiseptica phase I organism possesses a capsule and has the ability to agglutinate with K antiserum, although phase III organism lacks both. The present study examines the effect of sulfamonomethoxine (SMMX) on capsule formation of B. bronchiseptica. I also investigated whether or not the organisms possessed a capsule by bacterial agglutination with K antiserum. Three SMMX-resistant strains of B. bronchiseptica phase I organisms showed loss of agglutinability with K antiserum by culturing them at a higher concentration of 1.56 micrograms/ml of SMMX. These results indicated that capsule formation of SMMX-resistant B. bronchiseptica is inhibited by SMMX.     

Serum haptoglobin concentration in growing swine after intranasal challenge with Bordetella bronchiseptica and toxigenic Pasteurella multocida type D.

The acute phase reaction, in association with progressive atrophic rhinitis (AR), was monitored for 3 wk using serum haptoglobin (HPT) quantification in thirty-six, 15 kg swine after intranasal challenge with varying doses of Pasteurella multocida type D (toxigenic strain) and Bordetella bronchiseptica. The challenge doses were administered alone or in combination with pigs divided into 9 isolated treatment groups. Increasing doses of B. bronchiseptica were associated with lower serum HPT (P < 0.05), whereas increasing doses of P. multocida tended to increase serum HPT (0.05 < P < 0.10). Significant and positive correlation of mean HPT and AR score was found in these pigs; increased AR scores were associated with elevated mean HPT concentration (r = 0.41, P < 0.01). A significant interaction between P. multocida and B. bronchiseptica dose indicated that increasing the dose of B. bronchiseptica, for a fixed P. multocida dose, was associated with less AR (P < 0.05). The AR scores were greater in pigs given P. multocida, than B. bronchiseptica alone. These results indicate that a complex interaction between Pasteurella multocida and Bordetella bronchiseptica causes progressive atrophic rhinitis and alters serum HPT concentration in swine.