Use of supplemented Stainer-Scholte broth for the isolation of Bordetella pertussis from clinical material

Koenig, C. H. W. Van; Tacken, Angelika; Finger, H

doi:10.1128/jcm.26.12.2558-2560.1988

Cited by 20 publications

(5 citation statements)

References 10 publications

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“…Bordetella strains were grown at 37°C for 2 to 3 days on Bordet-Gengou (BG) agar plates (Becton, Dickinson) containing 1% Hipolypeptone (Nihon Pharmaceutical), 1% glycerol, 15% defibrinated horse blood, and 10 μg/mL ceftibuten. The bacteria grown on BG plates were suspended in Stainer-Scholte (SS) broth ( 40 , 41 ) to make a value of optical density at 650 nm (OD 650 ) of 0.1 ( B. pertussis ) or 0.02 ( B. bronchiseptica ) and incubated at 37°C with shaking for 16 h. The CFU was estimated from the OD 650 values of fresh cultures according to the following equation: 1 OD 650 unit = 3.3 × 10 9 CFU/mL. Escherichia coli was grown at 37°C on Luria-Bertani (LB) agar or in LB broth.…”

Section: Methodsmentioning

confidence: 99%

Survival of Bordetella bronchiseptica in Acanthamoeba castellanii

et al. 2023

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The pathogenic bacterium Bordetella bronchiseptica , which causes respiratory diseases in various mammals, exhibits distinct Bvg + and Bvg − phenotypes. The former represents the virulent phase, in which the bacteria express a set of virulence factors, while the role of the latter in the bacterial life cycle remains to be understood.

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Section: Methodsmentioning

confidence: 99%

Survival of Bordetella bronchiseptica in Acanthamoeba castellanii

et al. 2023

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“…To carry out the planktonic culture of B. pertussis , the biomass obtained after replating for 24 h on BGA was used as inoculum for the Stainer-Scholte ( 37 , 38 ) liquid medium supplemented with 1% casaminoacids (SSC). This inoculated medium was incubated at 36.5°C with shaking at 160 rpm until the optical density at 650 nm reached 1.2.…”

Section: Methodsmentioning

confidence: 99%

Pertussis Vaccine Candidate Based on Outer Membrane Vesicles Derived From Biofilm Culture

et al. 2021

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Outer membrane vesicles (OMV) derived from Bordetella pertussis—the etiologic agent of the resurgent disease called pertussis—are safe and effective in preventing bacterial colonization in the lungs of immunized mice. Vaccine formulations containing those OMV are capable of inducing a mixed Th1/Th2/Th17 profile, but even more interestingly, they may induce a tissue-resident memory immune response. This immune response is recommended for the new generation of pertussis-vaccines that must be developed to overcome the weaknesses of current commercial acellular vaccines (second-generation of pertussis vaccine). The third-generation of pertussis vaccine should also deal with infections caused by bacteria that currently circulate in the population and are phenotypically and genotypically different [in particular those deficient in the expression of pertactin antigen, PRN(-)] from those that circulated in the past. Here we evaluated the protective capacity of OMV derived from bacteria grown in biofilm, since it was observed that, by difference with older culture collection vaccine strains, circulating clinical B. pertussis isolates possess higher capacity for this lifestyle. Therefore, we performed studies with a clinical isolate with good biofilm-forming capacity. Biofilm lifestyle was confirmed by both scanning electron microscopy and proteomics. While scanning electron microscopy revealed typical biofilm structures in these cultures, BipA, fimbria, and other adhesins described as typical of the biofilm lifestyle were overexpressed in the biofilm culture in comparison with planktonic culture. OMV derived from biofilm (OMVbiof) or planktonic lifestyle (OMVplank) were used to formulate vaccines to compare their immunogenicity and protective capacities against infection with PRN(+) or PRN(-) B. pertussis clinical isolates. Using the mouse protection model, we detected that OMVbiof-vaccine was more immunogenic than OMVplank-vaccine in terms of both specific antibody titers and quality, since OMVbiof-vaccine induced antibodies with higher avidity. Moreover, when OMV were administered at suboptimal quantity for protection, OMVbiof-vaccine exhibited a significantly adequate and higher protective capacity against PRN(+) or PRN(-) than OMVplank-vaccine. Our findings indicate that the vaccine based on B. pertussis biofilm-derived OMV induces high protection also against pertactin-deficient strains, with a robust immune response.

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“…In addition, these authors demonstrated that PTX and FHA produced in the presence of CDX were immunochemically and biologically identical to those produced in CDX-free culture. Furthermore, CDX addition to growth medium facilitates the isolation of B. pertussis from clinical specimens significantly (47).…”

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confidence: 99%

Surface-associated filamentous hemagglutinin induces autoagglutination of Bordetella pertussis

et al. 1994

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Filamentous hemagglutinin (FHA) is a major adhesin produced by BordeteUla pertussis, the etiologic agent of whooping cough. FHA has been shown to be surface associated but is also secreted by virulent bacteria. Microscopic observations of lungs of mice infected with B. pertussis showed that the bacteria grow as clusters within the alveolar lumen. When B. pertussis was cultivated in vitro with chemically defined medium, bacteria grew as aggregates, mimicking growth observed in vivo. This aggregation was abolished by the addition of cyclodextrin (CDX) to the growth medium and depended on the production of FHA, because a mutant lacking the FHA structural gene failed to form aggregates in a CDX-free medium. Western blot (immunoblot) analyses revealed that, in the absence of CDX, FHA was attached to the bacterial surface and was not efficiently released into the growth medium. Hydrophobic chromatography of FHA showed that CDX drastically reduced the hydrophobicity of FHA, suggesting a direct binding of CDX to FHA, which was further supported by the partial protection of FHA from trypsin digestion in the presence of CDX. In addition, free FHA can interact in a CDX-inhibitable manner with solid phase-immobilized FHA. It can therefore be postulated that the B. pertussis aggregates are most likely due to direct FHA-FHA interaction. Bordetella pertussis, the etiologic agent of whooping cough, produces several virulence-associated adhesion factors such as

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Use of supplemented Stainer-Scholte broth for the isolation of Bordetella pertussis from clinical material

Cited by 20 publications

References 10 publications

Survival of Bordetella bronchiseptica in Acanthamoeba castellanii

Survival of Bordetella bronchiseptica in Acanthamoeba castellanii

Pertussis Vaccine Candidate Based on Outer Membrane Vesicles Derived From Biofilm Culture

Surface-associated filamentous hemagglutinin induces autoagglutination of Bordetella pertussis

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