Characterization of Bordetella bronchiseptica strains using phenotypic and genotypic markers

Thirty-five strains of Bordetella bronchiseptica, recovered primarily from pigs, rabbits, dogs, cats and humans, were characterized by phenotypic and genotypic markers. Biochemical typing only showed variation in the ability to reduce nitrate to nitrite. OMP profiles from virulent strains showed variations in the region of 85-95kDa, which lead us to describe five OMP-types alpha, beta, gamma, delta and varepsilon. Genotypic markers included the presence of IS1001, and polymorphisms in the flagellin gene (flaA) and pertussis toxin (PT) promoter region. The IS1001 was detected in 16 isolates (2 from humans and 10 from pigs) but was absent in rabbit isolates. The restriction profiles of the flaA gene allowed us to differentiate the strains into types A-C. The PT types were characterized by an RFLP assay and could be typed through patterns III-V. There was no apparent association between the flaA or PT types and the origin of the isolates. Eleven groups of isolates were identified on the basis of specific combinations of the analyzed markers. The combination of phenotypic and genotypic tests used could be useful in characterizing isolates and differentiating between certain clonal types of B. bronchiseptica.     

Crystal violet staining of Bordetella bronchiseptica colonies for differentiation of phase-I strains from variant strains in degraded phases.

After 2 days of growth on Brain heart infusion agar (BHIA) at 38 degrees C, phase-I colonies and degraded-phase colonies of Bordetella bronchiseptica could be differentiated by their ability to take up crystal violet (CV). Phase-I colonies in X mode, but not colonies in degraded phases (phases II, III, and rough) bound CV. Phenotypically-altered C-mode colonies (grown at 32 degrees C or lower temperatures) also lacked this ability. CV staining offers an easy method for the recognition of different colony types that appear identical when observed on BHIA.     

The differentiation of Bordetella (B.) bronchiseptica strains. 8. The protective behavior of B. bronchiseptica strains in mice in relation to the loss of K factors

K factors are phase-dependent, instable antigen components of B. bronchiseptica. Loss of K factors was induced by passaging on 10% horse blood agar. All K factors were not simultaneously eliminated. Cultures obtained from various passages were checked on white mice by Lienotox testing and in protectivity experiments. Culturing of B. bronchiseptica on 10% horse blood agar in Phase I probably had an adverse impact on expression of virulence-dependent factors. K factor expression in B. bronchiseptica was found to depend on both phase condition and culturing medium.     

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.     

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.     

猪波氏杆菌病的诊治

1998年 3— 7月份我县某集约化种猪繁殖场 60～ 90日龄生猪发生了一种慢性支气管肺炎为主要特征的传染病。临床上以咳嗽、喘气为主 ,尤其咳嗽较为明显 ,像咽喉或气管内有异物咳出来的连声长咳 ;对抗生素耐药性强 ,一旦发病病情顽固 ,很难彻底治愈 ;经流行病学、临床症状、剖检变化、病原分离、鉴别诊断等 ,确诊为猪败血波氏杆菌病。该病在我国生猪发病病例少见报道 ,介绍如下。1　流行情况　 1 998年 3月某集约化种猪繁殖场引进 40头长白母猪 ,推广经济杂交组合 ,改良生猪品种。不久猪场部分断奶仔猪和小架子猪出现咳嗽、喘气症状 ,经抗生素…     

猫源支气管败血波氏菌的分离与鉴定

支气管败血波氏菌是引起猫呼吸道疾病的主要病原之一,而对猫源支气管败血波氏菌的分离鉴定目前国内鲜有报道。本研究对上海市1例有咳嗽症状的德文猫鼻拭子样品进行呼吸系统相关病原PCR检测,并对鼻拭子样品进行细菌分离培养,对分离菌株采用K-B纸片法进行药敏试验,同时检测分离菌株对小鼠的致病性。结果显示：猫杯状病毒、猫疱疹病毒、猫衣原体、猫支原体均为阴性,支气管败血波氏菌为阳性,并分离到1株支气管败血波氏菌;分离菌株对头孢拉定、氨苄西林、强力霉素等9种抗生素耐药,对阿莫西林、卡那霉素、氟哌酸、复方新诺明、美罗培南等15种抗生素敏感;用分离菌株接种ICR小鼠,发现小鼠出现精神沉郁症状,但未出现死亡。结果表明：上海市猫群中存在支气管败血波氏菌感染,需关注免疫力低下人群被感染的风险;分离株对小鼠致病力不强,但对首选治疗支气管败血波氏菌病的强力霉素耐药,建议选用敏感药物进行防治。本研究为猫源支气管败血波氏菌病的诊断与防治提供了参考。     

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.     

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