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Bordetella bronchiseptica is a Gram-negative respiratory pathogen responsible for atrophic rhinitis and bronchopneumonia in swine. Several vaccines aimed at preventing B. bronchiseptica have been used, but a safe and efficient live vaccine for use in piglets remains elusive. In this study, we constructed an aroA-deleted B. bronchiseptica strain (QH0814) and evaluated its safety and protective efficiency in piglets. Lung lesion scores in QH0814-immunized piglets post-challenge were significantly lower than those in piglets immunized with the parent strain (P<0.05). Immunization with QH0814 induced a vigorous immune response, especially at the mucosal surface of the respiratory tract. IgA titers in bronchoalveolar lavage fluid (BALF) and serum were significantly higher in the QH0814-immunized group compared to the inactivated-vaccine-immunized group. Piglets immunized with QH0814 were better protected than those in the inactivated-vaccine and negative control groups. The clinical symptoms, histopathological changes and immune responses elicited in the piglets were recorded. The results of this study suggest that QH0814 was able to confer complete protection against B. bronchiseptica infection and could thus be used as a candidate attenuated live vaccine against B. bronchiseptica in piglets., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Between January 2003 and September 2008, 652 Bordetella bronchiseptica isolates were cultured from 3506 lung samples collected from pigs with respiratory disease. Over the 6-year period, the average isolation rate was 18.6%, making B. bronchiseptica the fourth most frequently isolated pathogenic bacterium from those lung samples. The isolation rates in different years and provinces ranged from 15.2% to 25.7% and 17.3% to 20.7%, respectively. There were significant influences of sampling month and pig age on bacterial isolation (P<0.05). Streptococcus suis (29.9%), Haemophilus parasuis (26.7%) and Escherichia coli (21.6%) were isolated most frequently in association with B. bronchiseptica. All 12 toxigenic Pasteurella multocida strains co-isolated with B. bronchiseptica from 63 cases of atrophic rhinitis were classified into serogroup D. The results suggest that B. bronchiseptica infection is highly prevalent in pig farms in China, and is often accompanied by co-infection with other bacteria., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Bordetella bronchiseptica is a Gram-negative respiratory pathogen that causes substantial disease in a variety of animals. Filamentous hemagglutinin (FHA) and pertactin are important attachment factors and protective immunogens, which serve as protective antigens in several animal models of infection with B. bronchiseptica. Here, we showed the efficacy of subcutaneous immunization of mice with a recombinant protein rF1P2, which consisted of the important immunodominant protective type I domain (F1) of FHA and the highly immunogenic region II domain (P2) of pertactin. Groups of mice tested, when challenged with different strains of B. bronchiseptica were fully protected, with long-lasting immunity to lethal B.bronchiseptica challenge, whereas mice immunized with Freund's adjuvant alone or PBS were not. In rF1P2-immunized mice, specific antibodies lasted for more than 120 days, and the IgG1/IgG2a ratio remained at a constant level till the end of the study. This suggests that rF1P2-induced a long-lasting balanced humoral immune responses and immunological memory in mice. rF1P2-specific antisera inhibited hemagglutination associated with full-length mature FHA. Furthermore, passive antiserum transfer from immunized animals completely protected naive mice from subsequent B. bronchiseptica challenge. These data may have implications for the development of safe and efficacious subunit vaccines for the prevention of bordetellosis, and may contribute to future acellular whooping cough vaccines.

Objective: We evaluated the efficacy of the recombinant pertactin (PRN)-specific active or passive immunization against Bordetella bronchiseptica (Bb) in an aerosol challenge model established by using BALB/c mice., Methods and Results: Mice, immunized subcutaneously with two doses of purified glutathione S-transferase (GST)-PRN protein mixed with an equal volume of Freund's adjuvant, produced robust PRN-specific IgG antibody activity. All 20 mice vaccinated with GST-PRN protein survived aerosol challenge with three times the 50% lethal dose (LD50) of virulent Bb HH0809 compared with 3 of 20 GST protein-treated controls and 4 of 20 PBS-treated controls that survived. Furthermore, we observed complete protection against intraperitoneal challenge with ten times the LD50 of virulent HH0809 strain in mice that were injected intraperitoneally with 0.5 ml rabbit anti-GST-PRN serum. No survivors were observed in mice that received either rabbit anti-GST serum or PBS alone., Conclusion: The recombinant PRN protein had strong immunogenicity against Bb infection.

Objective: We developed an indirect ELISA method for detecting Bordetella bronchiseptica (Bb) pertactin antibodies based on the recombinant pertactin protein expressed in Escherichia coli (DE3) strain., Methods and Results: The prn gene encoding Bb pertactin was fused to the downstream of glutathione S-transferase (GST) of pGEX-KG expression vector, resulting in the fusion expression plasmid pGEX-prn. SDS-PAGE showed that the GST-PRN fusion protein was expressed in high level in BL21 carrying pGEX-prn. The strong reactivity of the GST-PRN fusion protein, specifically with antiserum against porcine Bordetellosis caused by Bb HH0809, was identified by Western blot. The recombinant protein fragment of rPRN was purified from the GST-PRN fusion protein digested by protease thrombin with the purity of 93.1%. The rPRN-based indirect ELISA was developed for detecting antibodies against PRN. The ELISA could detect positive samples in experimentally infected pigs fourteen days post inoculation and the degree of sensitivity was over 4 times higher than the latex agglutination test with the coating antigen of killed Bb. Thirty-two point seven percent of positive samples were detected in 1,229 clinical samples while no false positive results were found in detecting 7 antisera against porcine bacterial diseases. Sera samples from two bordetellosis-positive pig fields were tested by the indirect ELISA method and the results indicated that pigs were infected by Bb during the nursery periods., Conclusion: The assay showed excellent specificity, sensitivity and reduplication, and can be useful for epidemiological survey and clinical diagnosis of swine bordetellosis.