An evaluation in pigs of Nobi-Vac AR and an experimental atrophic rhinitis vaccine containing P multocida DNT-toxoid and B bronchiseptica

The trial involved eight large white sows obtained from a closed experimental specific pathogen free herd. Four sows (two each for an experimental vaccine and for Nobi-Vac AR) were vaccinated twice (eight weeks and two weeks before parturition) with 2 ml of vaccine administered intramuscularly. Two unvaccinated sows were used as an infected control group and two unvaccinated sows served as an uninfected control group. Forty-six piglets (28 from vaccinated sows and 18 from unvaccinated sows) were challenged by intranasal instillation of Bordetella bronchiseptica at two days of age and Pasteurella multocida type D, dermonecrotic toxin at seven days of age. Among the infected control group some piglets died and there were clinical signs of pneumonia and severe turbinate atrophy. In the vaccinated groups the results showed that immunisation of the pregnant sows had provided a good level of antibodies, which were transmitted to their offspring. There was a significant reduction in the clinical signs and no lesions were observed in the group vaccinated with the experimental vaccine and only moderate atrophy of the turbinates in the Nobi-Vac AR group. B bronchiseptica and P multocida were never recovered from the lungs of the vaccinated groups and in the nasal cavities their frequency declined with age.     

Toxigenic type A Pasteurella multocida as a causative agent of nasal turbinate atrophy in swine.

Although no clinical signs of atrophic rhinitis (AR) were recognized in 2- and 5-week-old pigs, approximately 60% of 2- to 6-month-old pigs showed clinical signs of AR in an affected pig farm. None of the pigs had normal turbinate at slaughter. Bordetella bronchiseptica was not isolated from any of the pigs before onset and incipient stage of the outbreak (2-week to 2-month-old). Pasteurella multocida of capsular type D was not isolated from any of those pigs. However, toxigenic P. multocida of capsular type A was isolated from a number of the pigs immediately before onset and incipient stage of the outbreak. Thirty-six-day-old primary specific-pathogen-free pigs were inoculated intranasally with a toxigenic type A P. multocida isolated from a 5-week-old pig. Severe nasal turbinate atrophy was observed in those pigs which were necropsied at 3 weeks post-inoculation. This is the first report on outbreak of severe nasal turbinate atrophy induced by toxigenic type A P. multocida in Japan.     

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.     

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.     

Effects of intranasal inoculation with Bordetella bronchiseptica, porcine reproductive and respiratory syndrome virus, or a combination of both organisms on subsequent infection with Pasteurella multocida in pigs

OBJECTIVE: To determine effects of intranasal inoculation with porcine reproductive and respiratory syndrome virus (PRRSV) or Bordetella bronchiseptica on challenge with nontoxigenic Pasteurella multocida in pigs. ANIMALS: Seventy 3-week-old pigs. PROCEDURE: In experiment 1, pigs were not inoculated (n= 10) or were inoculated with PRRSV (10), P. multocida (10), or PRRSV followed by challenge with P. multocida (10). In experiment 2, pigs were not inoculated (n = 10) or were inoculated with B. bronchiseptica (10) or PRRSV and B. bronchiseptica (10); all pigs were challenged with P. multocida. Five pigs from each group were necropsied 14 and 21 days after initial inoculations. RESULTS: Pasteurella multocida was not isolated from tissue specimens of pigs challenged with P. multocida alone or after inoculation with PRRSV. However, in pigs challenged after inoculation with B. bronchiseptica, P. multocida was isolated from specimens of the nasal cavity and tonsil of the soft palate. Number of bacteria isolated increased in pigs challenged after coinoculation with PRRSV and B. bronchiseptica, and all 3 agents were isolated from pneumonic lesions in these pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Infection of pigs with B. bronchiseptica but not PRRSV prior to challenge with P. multocida resulted in colonization of the upper respiratory tract and tonsil of the soft palate with P. multocida. Coinfection with PRRSV and B. bronchiseptica predisposed pigs to infection of the upper respiratory tract and lung with P. multocida. Porcine reproductive and respiratory syndrome virus and B. bronchiseptica may interact to adversely affect respiratory tract defense mechanisms, leaving pigs especially vulnerable to infection with secondary agents such as P. multocida.     

Adherence of Bordetella bronchiseptica and Pasteurella multocida to porcine nasal and tracheal epithelial cells.

The ability of 19 different Bordetella bronchiseptica isolates and 25 Pasteurella multocida isolates to adhere in vitro to porcine nasal and tracheal epithelial cells was examined. It was found that B. bronchiseptica adhered well to upper respiratory tract cells. In contrast the number of P. multocida organisms which adhered was four to six times less than the number of B. bronchiseptica adherent organisms. This difference was statistically significant (p less than 0.0001). Both microorganisms adhered in greater numbers to nasal cells than to tracheal cells (p less than 0.005). The data indicated that B. bronchiseptica possesses a greater ability than P. multocida to attach to porcine upper respiratory tract cells.     

Interactions between Bordetella bronchiseptica and toxigenic Pasteurella multocida in atrophic rhinitis of pigs

Three strains of Bordetella bronchiseptica were compared for their ability to assist colonisation of the nasal cavity of gnotobiotic pigs by toxigenic Pasteurella multocida. Toxigenic P multocida (counted in nasal washings) colonised the cavity in large numbers in pigs previously infected with a cytotoxic phase I strain of B bronchiseptica (B58), whereas it colonised only in small numbers in those previously infected with B65, a phenotypic phase III variant of B58. Toxigenic P multocida colonised pigs infected with a non-cytotoxic phase I strain of B bronchiseptica (PV6) in fewer numbers than were seen in pigs infected with the cytotoxic phase I strain but in greater numbers than in pigs infected with the phase III strain. The turbinates of pigs infected with the cytotoxic phase I strain of B bronchiseptica and toxigenic P multocida were most severely affected and those in pigs infected with the non-cytotoxic phase I strain and toxigenic P multocida were moderately reduced in size. The turbinates of pigs infected with the phase III strain and toxigenic P multocida were slightly reduced in size except for one piglet whose turbinates were severely affected. Pigs infected with the non-cytotoxic phase I strain of B bronchiseptica alone showed no signs of atrophy and their turbinates were used to calculate reductions (per cent) in those infected with P multocida. The reduction (per cent) in size of turbinates and total numbers of P multocida isolated from the nasal washings of each pig were linearly related.(ABSTRACT TRUNCATED AT 250 WORDS)     

规模化猪场预防萎缩性鼻炎方法的比较试验

本试验对单独应用猪萎缩性鼻炎、多杀性巴氏杆菌二联苗和疫苗接种结合药物防制进行了效果比较。结果表明,7日龄首免1 mL/头,28日龄二免2 mL/头,可产生较强的免疫保护力。结合饲料中添加药物、哺乳仔猪药水滴鼻等药物防治措施,可使猪群的临床发病率由51.39％降至1.39％,生长肥育猪的综合性能指标得到较大改善。     

Experimental atrophic rhinitis in 2 and 4 month old pigs infected sequentially with Bordetella bronchiseptica and toxigenic type D Pasteurella multocida.

Experimental infections with Bordetella bronchiseptica and/or toxigenic type D Pasteurella multocida were studied in 2- and 4-month-old primary specific-pathogen-free pigs. None of the 2-month-old pigs inoculated with B. bronchiseptica or P. multocida alone developed turbinate atrophy. All the pigs inoculated with B. bronchiseptica (10(7) CFU/head) and P. multocida (10(9) CFU/head for 5 consecutive days) together, however, developed clinical and post-mortem signs of atrophic rhinitis (AR) similar to the naturally occurring disease. Slight to severe turbinate atrophy was observed in the 4-month-old pigs inoculated with B. bronchiseptica and P. multocida (at the same concentration as above) at necropsy.     

Expression of a truncated Pasteurella multocida toxin antigen in Bordetella bronchiseptica

Mild or subclinical respiratory infections caused by Bordetella bronchiseptica are widespread in pigs despite multiple control efforts. Infection with virulent B. bronchiseptica strains is a common risk factor in the establishment of toxin-producing strains of Pasteurella multocida in the nasal cavity of pigs leading to the disease, atrophic rhinitis (AR). This study was designed to explore the possibility of expressing a protective epitope of P. multocida toxin (PMT) in B. bronchiseptica to create single-component mucosal vaccine to control atrophic rhinitis in pigs. To achieve this, a P. multocida toxin fragment (PMTCE), that was non-toxic and protective against lethal challenge in mice, was cloned into a broad-host-range plasmid, PBBR1MCS2, and introduced into B. bronchiseptica by electroporation. The Pasteurella gene construct was placed under the regulatory control of a promoter region that was separately isolated from B. bronchiseptica and appears to be part of the heat shock protein gene family. B. bronchiseptica harboring the plasmid under antibiotic selection expressed the 80kDa PMTCE as determined by PAGE and Western blot with a PMT-specific monoclonal antibody. When introduced into the respiratory tracts of mice, B. bronchiseptica harboring the plasmid construct was reisolated in declining numbers for 72h post-inoculation. Antibody responses (IgM, IgA and IgG) to B. bronchiseptica were detected in serum and respiratory lavage, but PMTCE-specific antibodies were not detected. While further refinements of PMT expression in B. bronchiseptica are necessary, this study provides a basis for the development of a single-component, live-attenuated vaccine against atrophic rhinitis.     

Protection of piglets against atrophic rhinitis by vaccinating the sow with a vaccine against Pasteurella multocida and Bordetella bronchiseptica

The efficacy of a new vaccine against atrophic rhinitis in pigs was tested in the Netherlands and Denmark. The vaccine contained protein dO (a truncated Pasteurella multocida toxin which is immunogenic and non-toxic), inactivated Bordetella bronchiseptica whole cells, and an adjuvant. The sows were either vaccinated intramuscularly with 2 ml of the vaccine at six to eight and two to four weeks before expected farrowing or left unvaccinated as controls. All the piglets were challenged intranasally with B bronchiseptica when three to seven days old and with P multocida three to four days later. Pigs born to the vaccinated sows performed significantly better than pigs born to the control sows when judged on growth, average daily weight gain and snout scores. The challenge organisms were reisolated more frequently from the control pigs than from the pigs in the vaccinated group. The vaccinated sows and their progeny developed high titres of antibodies against B bronchiseptica and P multocida toxin.     

Induced atrophic rhinitis in rats

Infections with Bordetella bronchiseptica and Pasteurella multocida were inducted in newborn specific-pathogen-free rats. Turbinate atrophy was quantified by measuring the length of the osseous core of the ventral turbinates. Bordetella bronchiseptica readily colonized the nasal cavity. Inoculated rats developed serum agglutinating antibodies to B bronchiseptica. Turbinate atrophy, correlating with a severe inflammatory reaction, was observed after dual inoculation with B bronchiseptica or after a single inoculation when the inoculum contained relatively large numbers of B bronchiseptica. Pasteurella multocida only rarely colonized the nasal cavity, even after prior instillation of weak acetic acid solution or B bronchiseptica.     

Toxigenic Pasteurella multocida in rabbits with naturally occurring atrophic rhinitis

In a commercial rabbitry nasal swabs were taken from 36 animals with enzootic upper respiratory disease resembling porcine atrophic rhinitis. 35 Pasteurella multocida strains were isolated from 17 rabbits. Among 30 strains tested for dermonecrotic toxin production 3, derived from 3 animals, were positive in the guinea pig skin test. 15 Bordetella bronchiseptica strains were recovered from 14 rabbits. No toxigenic strains were found among 6 isolates tested using the same method.     

Clinical and pathological effects of the dermonecrotic toxin of Bordetella bronchiseptica and Pasteurella multocida in specific-pathogen-free piglets

The role of dermonecrotic toxin (DNT) of Bordetella bronchiseptica and Pasteurella multocida, purified by repeated chromatography in Sephacryl S-200 gel, in the pathogenesis of atrophic rhinitis (AR) of swine was studied bacteriologically, clinically and pathologically. Two-week-old specific pathogen-free (SPF) piglets were parenterally treated with 30 micrograms of DNT 3 times at 2-day interval and 7-week-old piglets were treated with 15 micrograms of DNT twice a week for 5 weeks. In 2- to 3-week-old piglets, both B. bronchiseptica DNT and P. multocida DNT produced nasal turbinate lesions with similar severity, characterized by damage of the cilia, epithelial metaplasia, intensive proliferation of osteoblasts, regressive changes, and diffuse osteocytic osteolysis. In 7- to 12-week-old piglets, treatment with B. bronchiseptica DNT failed to produce progressive changes in the nasal turbinates. Histopathological examination revealed osteogenic processes and osteoid synthesis besides the proliferation of osteoblasts and mild osteocytic osteolysis. Moreover, severe gross pathological lesions developed in the stomach, liver, kidneys, and lymphoid organs. The piglets' appetite and body weight gain gradually decreased during the DNT treatment and in the last week when the toxic signs appeared. Treatment of 7- to 12-week-old piglets with P. multocida DNT resulted in progressive AR. Histopathologically, diffuse osteocytic osteolysis was observed in the nasal turbinates. Neither clinical signs nor pathological lesions of the visceral organs developed in these piglets. The authors emphasize that the DNT of B. bronchiseptica basically differs from that of P. multocida in biological properties, though there are certain similarities between the DNTs.     

Prevention and treatment of atrophic rhinitis in pigs with Getroxel, chlorquinaldol and oxytetracycline.

The sensitivity of ten Bordetella bronchiseptica and ten Pasteurella multocida strains, each isolated from cases of atrophic rhinitis (AR), was examined in tube dilution test. Getroxel, chlorquinaldol and oxytetracycline and the former two ones combined with trimethoprim inhibited the growth of both species in vitro. The minimum inhibitory and the minimum bactericidal concentration was less than 0.5 microgram/ml. When efficacy was tested in SPF in the group fed a combination of Getroxel, chlorquinaldol and oxytetracycline (60 mg, 240 mg and 360 mg/kg of feed, respectively), P. multocida disappeared from the nasal cavity by the end of a 30-day treatment. B. bronchiseptica was reisolated in low numbers from 2 out of 9 piglets. The daily body mass gain was by 7.9% higher and the feed conversion rate was by 19% better than in the control group. After slaughter, only mild signs of AR were seen in 3 out of 9 piglets treated with the above-mentioned drug combination, while in the control group severe lesions were observed in 8 out of 9 pigs. In treated commercial herds P. multocida disappeared from the nasal cavity of the piglets by the end of the treatment (42nd day of life), but the B. bronchiseptica strains could not be completely eliminated. Due to the treatment, mortality between 2 and 6 weeks of age decreased by 0.8-7.6%. Daily body mass gain was, on the average, 16.4% higher, the amount of feed needed for 1 kg body mass gain was by 15.3% lower and the duration of fattening was by 30.8 days shorter than in the control groups.     

Associations between pathogens in healthy pigs and pigs with pneumonia

The aim of this study was to evaluate the associations between different pathogens in the development of pneumonia and bronchopneumonia in pigs. Samples of bronchoalveolar lavage fluid from 100 pigs showing no clinical signs and 239 pigs with clinical signs of respiratory disease were examined for Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, US-type porcine respiratory and reproductive syndrome virus (PRRSV), EU-type PRRSV, porcine circovirus type 2 (pcv-2), influenza virus type A, alpha-haemolytic Streptococcus species, beta-haemolytic Streptococcus species, Pasteurella multocida, Bordetella bronchiseptica, Haemophilus parasuis and Actinobacillus pleuropneumoniae. These potential pathogens were detected more frequently in the pigs with respiratory problems than in the pigs with no clinical signs. pcv-2 and alpha-haemolytic streptococci were the pathogens most frequently detected; A pleuropneumoniae was isolated in only two cases. There were more often associations between the organisms in the pigs with clinical signs than in the healthy pigs. In particular, alpha-haemolytic streptococci and M hyopneumoniae were both associated with the presence of M hyorhinis, EU-type PRRSV, P multocida and B bronchiseptica, and alpha-haemolytic streptococci also occurred more often in pigs that were already infected with other pathogens. P multocida and B bronchiseptica were both significantly associated with M hyopneumoniae, alpha-haemolytic streptococci, EU-type PRRSV and US-type PRRSV.     

An agglutination test for the detection of Bordetella bronchiseptica infection in swine.

An agglutination test with the use of formalin-killed antigen of the cell carrying the capsule was developed and used for the detection of antibody in swine naturally infected with Bordetella bronchiseptica. Under optimum antigen concentration and reaction temperature 210 or 60% of 342 serum samples tested from 42 conventional swine herds were positive for Bordetella infection. In contrast, only 34 or 10% of 342 nasal swabs from the same animals were positive for Bordetella by culture technique. The test was relatively free of cross-reactivity to related organism. However, 2.7 and 13.0% of sera from growing pigs and mature hogs, respectively, reacted with antigen of Pasteurella multocida. Because of this, only agglutinin reactions in 1:20 dilutions or higher to Bordetella were considered positive. The bulk of the antibody activity of selected sera tested from various age ranges of swine was mercaptoethanol sensitive, suggesting that serum antibody in Bordetella infection may be associated with immunoglobulin IgM. Because of the high agglutinability and stability of formalin-killed antigen the test may be useful as an auxiliary aid for the diagnosis of Bordetella infection where the organism cannot be identified by culture means.     

Induction of nasal turbinate atrophy in germ-free pigs, using Pasteurella multocida as well as bacterium-free crude and purified dermonecrotic toxin of P multocida

To establish the role of the dermonecrotic toxin (DNT) of Pasteurella multocida in the cause and pathogenesis of atrophic rhinitis, germ-free pigs were inoculated with several strains of P multocida, crude DNT, or purified DNT. In some experiments, the aforementioned inocula were combined with Bordetella bronchiseptica. All DNT-producing P multocida strains induced severe turbinate atrophy. Histologic examination of the remnants of the nasal turbinates revealed intact, but undulated, ciliated epithelium and numerous osteoclasts. Inflammation was minimal or absent. A DNT-producing B bronchiseptica strain induced only mild turbinate atrophy. The lesions were characterized histologically by loss of cilia and ciliated cells and by an infiltration of predominantly mononuclear cells. Bone formation seemed impaired. Turbinate lesions were most severe in pigs infected with a combination of B bronchiseptica and a DNT-producing P multocida strain. Intranasal administration of sterile DNT-containing culture filtrate of P multocida or purified DNT of P multocida did not result in turbinate atrophy. In contrast, turbinate atrophy developed when these preparations were injected IM or when intranasal administration of DNT was preceded by inoculation of B bronchiseptica.     

Evaluation of an atrophic rhinitis vaccine under controlled conditions.

A vaccine containing inactivated cultures of Bordetella bronchiseptica, toxigenic Pasteurella multocida type D and dermonecrotic P multocida type D toxoid in an oil-in-water adjuvant was given to seven sows, with seven others acting as controls. Half the piglets in each litter were exposed intranasally when four days old to B bronchiseptica and when eight days old to toxigenic P multocida type D. There was considerably less sneezing in the litters of the vaccinated sows and when the piglets were 10 weeks old, only 18 per cent had deformed snouts compared with 74 per cent in the litters of the control sows. The average liveweight gain of the piglets born to vaccinated sows was significantly better (P less than 0.05) between two and 10 weeks of age than that of the piglets born to unvaccinated sows, although there were no significant lower respiratory tract lesions in either group. The conchal atrophy scores were significantly lower (P less than 0.001) in the piglets from the vaccinated sows and were negatively correlated (r = -0.37) with increasing liveweight gain. In the liters of the vaccinated sows, P multocida was not isolated from the nasal passages of the in-contact piglets and from only 7 per cent of those deliberately exposed compared with 65 per cent and 79 per cent, respectively, in the litters of the control sows. P multocida was isolated post mortem from the tonsils of 23 per cent of the piglets of vaccinated sows and from 87 per cent of those from unvaccinated sows.     

The synergistic activity of tiamulin and chlortetracycline: in-feed treatment of bacterially complicated enzootic pneumonia in fattening pigs

The antibacterial effects of a combination of tiamulin and chlortetracycline in vitro against a number of field isolates of Pasteurella multocida, Haemophilus pleuropneumoniae and Bordetella bronchiseptica were examined. There was a marked synergism between the two antibiotics against all eight isolates of P multocida, against seven of nine isolates of H pleuropneumoniae and against the single strain of B bronchiseptica tested. Two field trials were carried out on a herd with a history of complicated enzootic pneumonia where the presence of Mycoplasma hyopneumoniae and P multocida had been established and subsequently the presence of H pleuropneumoniae was discovered. Feed containing tiamulin at 100 ppm combined with chlortetracycline at 300 ppm was given for seven days to pigs affected with pneumonia, and the results were compared with untreated controls and pigs receiving chlortetracycline at 300 ppm. There was a follow-up observation period of three weeks when all groups received unmedicated feed. During the medication period the combination treated groups showed a statistically significant increase in average daily weight gain of 156 g (20.4 per cent) and in feed conversion efficiency of 0.576 (20.8 per cent) and a numerical improvement in average disease score in comparison with the untreated controls. These improvements were approximately double those observed in the groups treated with 300 ppm chlortetracycline which showed improvements of 93 g (12.2 per cent) in average daily gain and 0.301 (10.9 per cent) in feed conversion efficiency. During the following three weeks most of the initial gains were lost, probably owing to the reinfection of the treated groups by the untreated controls.