Absence of mouse-lethal toxins in Australian isolates of Bordetella avium

A total of 18 Australian isolates of Bordetella avium and seven reference B. avium strains from Europe and the USA were tested for the presence of a mouse-lethal toxin. Five of the seven reference strains of B. avium, but none of the 18 Australian isolates, produced the toxin.     

Lack of protection against Bordetella avium in turkey poults exposed to B. avium-like bacteria

Six laboratory experiments were designed to determine whether poults infected with the nonpathogenic Bordetella avium-like (BAL) bacteria would develop immunity to B. avium (BA), the causative agent of turkey coryza. The BAL bacteria were isolated from poults given that organism, but few colonies were observed by 3 weeks postexposure. No serum-agglutinating antibody to the BAL bacterium was detected in poults exposed to that organism. Poults exposed to BAL bacteria either once or twice at different ages were not protected from infection or disease following experimental challenge between 1 and 7 weeks of age with pathogenic BA.     

Antimicrobial susceptibility testing of Australian isolates of Brachyspira hyodysenteriae using a new broth dilution method.

The antimicrobial susceptibilities of 76 field isolates of Brachyspira hyodysenteriae from different states of Australia were tested in a newly developed broth dilution procedure. The antimicrobial agents used were tiamulin, valnemulin, tylosin, erythromycin, lincomycin and clindamycin. The results from the broth dilution susceptibility testing of 39 of the isolates were compared with results obtained for the same isolates using the agar dilution method. Amongst the isolates tested by broth dilution, 17 were from three farms and had been collected over a number of years. Their pulsed field gel electrophoresis pattern previously had been determined. The broth dilution technique was simple to use, less labor intensive than agar dilution, and gave clear end points. The results obtained using the two methods generally corresponded well, although in a few cases the MIC obtained by broth dilution were lower than those with agar dilution. For the 76 isolates tested by broth dilution, the MIC(90) (mg/l) was: tiamulin, 1; valnemulin, 0.5; tylosin>256; erythromycin>256; lincomycin, 64 and clindamycin, 16. Only minor differences in susceptibility patterns were found amongst isolates from different Australian states. Over all the isolates, and also amongst the isolates obtained from different years on the three farms, there was no trend for the susceptibility of the isolates to alter with time.     

Pathogenicity of Australian isolates of Haemophilus paragallinarum and Haemophilus avium in chickens

Twenty-seven Australian avian Haemophilus isolates were tested for their ability to cause infectious coryza in specific pathogen-free chickens. All 15 isolates, identified as H. paragallinarum, produced infectious coryza, whereas all 12 H. avium isolates were nonpathogenic, but spread to in-contact chickens.     

Isolation and characterization of Bordetella avium plasmids

Experiments were conducted to study the plasmids of Bordetella avium, B. avium-like, and B. bronchiseptica isolates from turkeys and the plasmids of the Art-Vax commercial vaccine strain. Plasmids were observed in 6 of 20 B. avium isolates, in 6 of 20 B. avium-like isolates, in all 5 B. bronchiseptica isolates, and in the Art-Vax strain. Plasmids of B. avium correlated with resistance to antibiotics but not with pathogenicity, hemagglutination of guinea pig erythrocytes, or expression of pili.     

Isolation of Bordetella avium from poultry

Four Australian isolates of Gram-negative nonfermentative bacteria obtained from poultry were compared with reference strains of Bordetella avium, Bordetella bronchiseptica and Alcaligenes faecalis. The Australian isolates were identified as Bordetella avium. A routine procedure for the identification of this recently recognised poultry pathogen is described.     

Antimicrobial susceptibility of Bordetella bronchiseptica isolates from cats and a comparison of the agar dilution and E-test methods

One hundred and fifty-two predominantly feline isolates of Bordetella bronchiseptica were tested for their susceptibility to seven antimicrobial agents using an agar dilution method. The majority of isolates tested by the agar dilution method were resistant to trimethoprim (MIC₉₀ 500 μg/ml) and ampicillin (MIC₉₀ > 32 μg/ml) but sensitive to tetracycline, doxycycline and enrofloxacin (MIC₉₀ 2 μg/ml for all three agents). The isolates showed a spectrum of susceptibility to sulphadiazine and clavulanate potentiated amoxycillin. The MIC's of twenty-nine of the 152 isolates were then compared for five of the antimicrobial agents using the E-test (AB Biodisk, Sweden), a recently introduced method for measuring the MIC's of antimicrobial agents based on the diffusion of a pre-defined antibiotic gradient from a plastic strip. Comparisons with the E-test demonstrated an overall agreement (±1 log₂ dilution) with the agar dilution method of 79.4% and an agreement within ±2 log₂ dilutions of 96.2%.     

DNA fingerprinting of Australian isolates of Mycobacterium avium subsp paratuberculosis using IS900 RFLP

OBJECTIVES: To evaluate additional restriction enzymes for IS900 RFLP of Mycobacterium avium subsp paratuberculosis and examine the genetic diversity among Australian isolates for epidemiological studies of Johne's disease. DESIGN AND PROCEDURE: Seventy-one isolates of M paratuberculosis from cattle, sheep, goat, alpaca and rhinoceros in six Australian States and the Northern Territory, reference strains and reference DNA from previously characterised strains were tested for genetic variation. Bst EII, Pvu II and Pst I restriction enzymes were used, and four others (Bam HI, Alu I, Xho I and Dra I) were assessed for their ability to detect polymorphisms. Multiple isolates from some animals were tested. RESULTS: Bam HI, was the most effective enzyme for identifying polymorphisms (12 types), followed by Bst EII (11 types). Both Pvu II and Pst I were relatively ineffectual. Fifteen different types were identified, 12 in clinical isolates. Most isolates were cattle (C) strains and fell into the C1 (n = 28) and C3 (n = 32) groupings. All isolates from alpaca were type C1, and bovine isolates were commonly C1 (n = 15) or C3 (n = 28). All of the sheep were infected with sheep (S) strains; no S strains were identified in cattle. Two of six isolates from one animal had single band differences. CONCLUSION: The epidemiological features of M paratuberculosis in Australia are similar to those reported in New Zealand, where cattle and sheep are commonly infected with different strains. However, because of the lack of polymorphism identified within the major groups, it is unlikely that DNA fingerprinting will have a significant role in epidemiological studies of Johne's disease, unless an unusual strain in being studied.     

Antimicrobial susceptibility patterns and sensitivity to tulathromycin in goat respiratory bacterial isolates

Bacterial pneumonia is a common and often life-threatening respiratory problem in both meat and dairy goats. Options for approved antibiotic therapy in goats to combat these bacterial infections are severely limited and frequently drugs must be used in an extra-label manner. Tulathromycin, a triamilide macrolide antimicrobial drug shown to be effective against swine and cattle respiratory bacterial agents, has been identified as a potentially useful drug in caprines. The present study was conducted to determine the susceptibility of recognized bacterial respiratory pathogens to commonly prescribed antimicrobials, with a particular emphasis on the efficacy of tulathromycin against these agents. Minimum inhibitory concentration (MIC) testing using microbroth dilution was performed on a collection of 45 Mannheimia haemolytica, 11 Pasteurella multocida, and 11 Bibersteinia trehalosi isolates from the lungs of goats with clinical pneumonia. To further characterize efficacy of tulathromycin against these pathogens, minimum bactericidal concentration (MBC) testing and kinetic killing assays were conducted. Most isolates were susceptible to the antimicrobials tested; however, increased resistance as demonstrated by higher MIC values was seen in all species to penicillin, in P. multocida to sulfadimethoxine, and in B. trehalosi to the tetracyclines. All isolates were susceptible to tulathromycin, which demonstrated a high killing efficiency in both bactericidal assays. Results of this study indicate that most goat pneumonic bacterial pathogens remain susceptible to commonly prescribed antibiotics, although some evidence of resistance was seen to certain drugs; and that tulathromycin is highly effective against goat respiratory pathogens which could make it a valuable medication in this species.     

直接荧光抗体法检测禽波氏杆菌

制备了兔抗禽波氏杆菌特异性荧光抗体并建立了直接荧光抗体检测方法 ,对禽波氏杆菌在人工感染雏鸡内的致病机理作了初步研究。结果 ,该荧光抗体只与其相应菌株发生特异性荧光反应 ,而不与其他病原菌株发生反应。对人工感染发病雏鸡的检测结果表明 ,禽波氏杆菌主要定植于上呼吸道黏膜 ,并造成损害。该技术具有简便、快速、敏感和特异性强等优点     

Ultrastructural pathology of Bordetella avium infection in turkeys

One-day-old turkeys were infected intranasally with Bordetella avium, and tracheas were examined by scanning and transmission electron microscopy at 1 to 5 weeks post-inoculation (PI). The predominant ultrastructural lesions were progressive loss of ciliated epithelium with replacement by nonciliated cells, bacterial colonization of ciliated cells, membrane-bound crystalline inclusions in cytoplasma of epithelial cells, depletion of mucous granules, and distortion of tracheal rings and the mucosal surface. Tracheal surface exudates consisted of mucus, necrotic cells, heterophils, and fibrin. Ciliated cells were replaced by immature cuboidal cells characterized by abundant rough endoplasmic reticulum with small numbers of electron-dense mucous granules in the apical cytoplasm. Bacterial surfaces were rough and contained numerous pleomorphic, knob-like structures, 20-50 nm in diameter. Other changes included enlarged mucosal gland openings, cell extrusion marks, pleomorphic microvilli, and cells with small numbers of short cilia.     

Antimicrobial susceptibility testing of Spanish field isolates of Brachyspira hyodysenteriae

This study is the first conducted in Spain to evaluate antimicrobial susceptibility of field isolates of Brachyspira hyodysenteriae. One hundred and eight isolates of the bacterium, recovered from different Spanish swine farms between 2000 and 2007, were investigated. The minimum inhibitory concentrations (MIC) of erythromycin, tylosin, tiamulin, valnemulin, clindamycin and lincomycin were determined using a broth microdilution technique. Most of the isolates showed poor susceptibility to erythromycin (MIC₉₀ > 256 μg/ml), tylosin (MIC₉₀ > 256 μg/ml), clindamycin (MIC₉₀ > 4 μg/ml) and lincomycin (MIC₉₀ = 128 μg/ml). Reduced susceptibility to tiamulin and valnemulin was observed with a MIC > 2 μg/ml in 17.6% and 7.41% of the B. hyodysenteriae isolates, respectively. Moreover, a survival analysis permitted the detection of an increasing trend in the MIC values for almost all the antimicrobials used in the treatment of swine dysentery when comparing recent isolates (from 2006 to 2007) with those recovered in earlier years (between 2000 and 2004).     

Development of an enzyme-linked immunosorbent assay for Bordetella avium

A Bordetella avium enzyme-linked immunosorbent assay (ELISA) was developed to detect serum antibodies in 1-day-old poults, experimentally infected turkeys, and naturally infected turkeys. The optimized procedure included use of a suspension of whole bacteria coated onto plastic microtiter plates, a 1:200 serum dilution, a 1:3200 dilution of commercially available goat anti-turkey IgG (heavy and light chain) conjugated with horseradish peroxidase, and 0.04% orthophenylenediamine as substrate. A sample/negative (S/N) ratio method of analysis was used to estimate antibody titer from absorbance values. The regression equation used to estimate antibody titers was derived from the testing of naturally infected turkey sera. The equation was derived by plotting the log10 titer of the sera against the S/N ratio at a 1:200 serum dilution. The ELISA was an effective method for detecting antibody to B. avium, and the procedure should prove useful for laboratories equipped for high-volume ELISA testing.     

Filamentous forms of Bordetella avium: culture conditions and pathogenicity.

Difficulties in preparing suitable antigen for the microagglutination (MA) test led to the discovery of a filamentous form of Bordetella avium. Broth media high in nutrients, vigorous shaking, and incubation at 37 C appeared to promote the development of filamentous forms of the organism. Peptone broth did not induce the development of filamentous forms. Eleven different isolates having both smooth and rough colony types were tested and observed to form filaments of various lengths. Filamentous forms of B. avium hemagglutinated guinea pig erythrocytes were motile, had pili and flagella, and were stable up to the fourth or fifth passage in broth media, at which time a predominance of typical coccobacillus organisms were observed. Filamentous forms of B. avium originating from smooth-colony types were pathogenic in turkey poults, whereas the filamentous forms of B. avium originating from rough-colony types were not pathogenic.     

应用23S rRNA对禽波氏杆菌分离株的进化分析

通过PCR扩增分离保存的8株禽波氏杆菌、3株兔支气管败血波氏杆菌及1株猪支气管败血波氏杆菌菌株的23S rRNA基因的片段(710bp),分别克隆到载体pMD18-T后测序。利用BLAST工具进行同源性搜索;用DNAStar分析软件进行同源核苷酸序列的多重比较分析并构建禽波氏杆菌菌株的系统生物进化树。结果显示,8株禽波氏杆菌核苷酸序列之间同源性为99.2%～99.7%,与GenBank中收录的NC_010645株禽波氏杆菌核苷酸序列同源性为92.4%～92.5%,与兔支气管败血波氏杆菌和猪支气管败血波氏杆菌的核苷酸序列同源性均为98.5%～99.2%。国内分离的禽波氏杆菌菌株和支气管败血波氏杆菌菌株均与国外分离菌株在遗传基因上有一定的差异。结果表明,23S rRNA序列分析可以作为鉴定禽波氏杆菌的一种快速简便的方法。     

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.