Antibiotic susceptibility of canine Bordetella bronchiseptica isolates

The antimicrobial sensitivities of 78 recent (1995-1998) canine isolates of Bordetella bronchiseptica from 13 separate sources were determined. Minimum inhibitory concentrations were assessed using the E-test method or by agar dilution. All 78 isolates were sensitive to tetracycline, doxycycline, enrofloxacin, and amoxycillin/clavulanic acid; the majority were sensitive to ampicillin (63/78; 81%), trimethoprim (57/78; 73%), and sulphadiazine (63/78; 81%). Plasmids were detected in 14 out of the 24 isolates tested. There was no correlation between the presence of plasmids and antibiotic resistance, but there was some correlation between the presence of plasmids and the origin of the isolates. Three sizes of plasmid were found: 20, 14, and 5.5 kb. Eight of the isolates contained all three plasmids, the remainder one or two, Thirteen isolates demonstrated beta-haemolysis, of which six produced a soluble haemolysin. Except for one isolate, haemolysin production correlated with plasmid carriage. Pulsed-field gel electrophoresis showed that all except one isolate could be grouped in the same genotype. Within this genotype isolates could be divided into three subtypes, generally corresponding to their place of origin.     

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.     

Studies on the antigenic relationships of six adherent isolates of Bordetella bronchiseptica

Six isolates of Bordetella bronchiseptica recovered from swine with atrophic rhinitis were studied. All hemagglutinated swine red blood cells, autoagglutinated in saline and showed fimbriae by electron microscopy. Hyperimmune sera against each were produced in rabbits and the antigenic relationships between the isolates were studied by cross-absorption and by the determination of the cross-reactivity indices of pairs of sera. Three isolates seemed to be identical by both methods, while 2 others showed close antigenic relationships. Hemagglutination titers with heterologous antigens and cross-reactivity indices greater than 0 suggest some degree of cross-immunity among the isolates studied, even when antigenic heterogeneity was demonstrated.     

Detection of type III secretion system genes in animal isolates of Bordetella bronchiseptica

A cosmid clone bank of Bordetella bronchiseptica genomic DNA was screened for the presence of type III secretion (TTS) genes using a probe derived from the TTS system genes of Ralstonia solanacearum. A 3.35kb PstI fragment, sub-cloned from a hybridising cosmid clone, was sequenced and found to contain a 97bp overlap with the previously reported B. bronchiseptica bscIJKLNO TTS gene cluster. DNA and predicted protein homology analysis suggests that a bscPQRST cluster lies immediately downstream of bscIJKLNO. A PCR amplification assay indicated that the bscT locus was present in 27 B. bronchiseptica animal isolates tested (100%). Dot-blot DNA hybridisation using probes for bscT and bscP confirmed the presence of these loci in six canine isolates associated with a variety of clinical signs. Although TTS has been implicated in the pathogenicity of B. bronchiseptica, it is likely that different clinical manifestations may be due to variations in gene expression or host factors, rather than the absence or presence of TTS genes.     

猪支气管败血波氏杆菌PCR诊断方法的建立及应用

根据支气管败血波氏杆菌（Bordetella bronchiseptica）鞭毛蛋白基因的上游序列设计了1对引物flal和fla2，扩增出大小为237bp的目的基因片段，建立了快速检测支气管败血波氏杆菌的PCR方法。特异性和敏感性试验表明，该方法对大肠埃希氏菌、金黄色葡萄球菌和D型多杀性巴氏杆菌均无交叉性反应；最低能够检出112．5pg的模板DNA。用该PCR法检测了从延边州采集的48份表现不同临床症状的猪鼻黏液；结果，检出支气管败血波氏杆菌阳性42例，阳性率为87．5％。同时与传统的细菌检查法、微量凝集法进行了比较；结果显示，该PCR法的检出率是细菌检查法的5．25倍，是微量凝集法的1．75倍。     

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)     

Detection of urease-negative Bordetella bronchiseptica from the field

Four urease-negative Bordetella bronchiseptica isolates originating from pigs were examined by phenotypic and molecular methods. The phenotypic properties of the isolates were in harmony with the data of the literature, except for the lack of urease activity in conventional tube test, API 20 NE and Diatabs? assays. Using genotypic methods, the urease-negative isolates did not differ from the urease-positive reference strain. They were positive in species-specific and ureC PCR, and all strains showed uniform bands in PCR-RFLP studies of flaA genes. The reason for the lack of urease activity, a characteristic considered species specific for B. bronchiseptica, needs to be studied further. The finding underlines the significance of genotyping when the phenotypic identification of B. bronchiseptica seems questionable.     

Prevalence and in-vitro antimicrobial sensitivity of Bordetella bronchiseptica in the nasal cavity of pigs

The prevalence of Bordetella bronchiseptica in the nasal cavity of pigs and the in-vitro sensitivity of isolates to a variety of antimicrobial agents was investigated. B.Bronchiseptica was recovered from 372 nasal swabs collected from 1000 (37.2%) pigs slaughtered at 20-30 weeks old at an abattoir. The swabs were collected from groups of 5-206 pigs derived from 25 herds. All isolates tested against bacitracin, clindamycin, furazolidone, penicillin, spectinomycin, streptomycin and tylosin were found to be resistant. Of the 372 isolates tested against ampicillin and erythromycin 22 (6%) were sensitive to the former and 365 (98%) were moderately sensitive to the latter, the remainder were resistant. All isolates tested against neomycin and tetracycline were sensitive and with few exceptions, (2%), they were also sensitive to chloramphenicol. Overall, 259 of the 372 (70%) isolates were sensitive to sulphonamides, identical results being obtained with sulphadiazine, sulphafurazole and a trimethoprim-sulphamethoxazole combination. An association between in-vitro resistance to sulphanomides and extensive use of this group of drugs was demonstrated on three of eight farms investigated.     

Strain- and growth condition-dependent variability in outer membrane protein expression by Bordetella bronchiseptica isolates from dogs.

OBJECTIVE: To characterize strain-dependent and growth condition-dependent variability in outer membrane protein (OMP) expression of Bordetella bronchiseptica isolates from dogs and evaluate the systemic immune response to OMP of B bronchiseptica among infected dogs. SAMPLE POPULATION: 8 strains of B bronchiseptica isolated from dogs, including a historic reference strain, 2 commercially available vaccine strains, and 5 field strains, and serum samples collected from 3 specific-pathogen-free (SPF) dogs before and 1 month after infection with B bronchiseptica. PROCEDURE: OMP were isolated from cultures in the late exponential phase of growth and compared among strains and, within strains, among growth conditions by means of polyacrylamide gel electrophoresis and immunoblotting. Serum samples were probed with OMP from 1 of the field strains. RESULTS: Strain-dependent variability in OMP profiles and growth condition-dependent and strain-dependent variability in expression of filamentous hemagglutinin (FHA) and pertactin was found, along with heterogeneity of the pertactin proteins produced by these B bronchiseptica strains. All 3 SPF dogs seroconverted to proteins with estimated molecular masses of 200 and 66 kDa, suggesting that FHA and pertactin were involved in the immunologic response of these dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that there is growth condition and strain variability in expression of OMP, FHA, and pertactin proteins produced by B bronchiseptica. This information could be useful in the improvement of vaccines for prevention of bordetellosis in dogs.     

Bordetella bronchiseptica infection in the cat

Feline Bordetella bronchiseptica infection had received little consideration until recent years when it has been increasingly documented in association with respiratory disease. This article reviews current knowledge on the organism; its epidemiology, pathogenesis, and clinical, diagnostic and therapeutic features.     

动物园野生动物支气管败血波氏杆菌的分离鉴定与系统发育树的构建

采用Biolog和16SrRNA基因序列分析法对分离自北京动物园的8株支气管败血波氏杆菌进行了鉴定。菌株经纯化培养,用Biolog微生物鉴定系统进行了鉴定,结果表明,8株菌株为支气管败血波氏杆菌。菌株经纯化培养,采用菌落PCR方法进行16SrRNA基因序列扩增,扩增产物纯化后直接进行测序。序列经人工校对后用ClustalX1.83软件进行比对分析,最后用Mega3.1软件构建系统发育树,系统发育分析结果表明,8株菌株与支气管败血波氏杆菌DSM10303的同源性达到了99.9%,并在同一个分支内。     

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.     

兔波氏杆菌病的诊断与综合防治

兔波氏杆菌病是由支气管败血波氏杆菌(B.bronchiseptica)引起的兔的呼吸道传染病。该病以发生慢性鼻炎和支气管肺炎为主要特征,近几年来,在我省的多个兔场中常有发生。本文对某兔场波氏杆菌病的诊断及综合防治措施简报如下。1 发病情况及临床症状 广州某兔场饲养近1 000只种兔和肉兔,2001年1月中旬,部分兔只出现打喷嚏、咳嗽、流鼻涕