Laboratory Maintenance of Bordetella pertussis

Abstract

The causative agent of the respiratory disease whooping cough, Bordetella pertussis, is a nutritionally fastidious microorganism but can be grown with relative ease in research laboratories. Stainer‐Scholte synthetic broth medium and Bordet‐Gengou blood agar both support growth of B. pertussis and are commonly used. B. pertussis prefers aerobic conditions and a temperature range of 35° to 37°C. Appropriate laboratory safety protocols are required to prevent the generation of aerosols, which could potentially spread this highly infectious agent. Curr. Protoc. Microbiol. 15:4B.1.1‐4B.1.9. © 2009 by John Wiley & Sons, Inc.

A rapid method for capture and identification of immunogenic proteins in Bordetella pertussis enriched membranes fractions: A fast-track strategy applicable to other microorganisms

“…Briefly, the cells were disrupted in a ThermoElectron (Waltham, MA) French press with one passage at 20000 psi. Unbroken cells were removed by centrifugation at 8000 ×g for 20 min at 4°C [14]. The supernatant was diluted in ice-cold 0.1M sodium carbonate (pH 11) to a final volume of 50 mL and stirred slowly for 2 h at 4°C.…”

“…Briefly, Tohama I Bp was grown and prepared as previously described [14]. The bacterial strain was grown on BordetGengou agar and then subcultured into Stainer-Scholte (SS) liquid medium.…”

A versatile, non genetically modified organism (GMO)‐based strategy for controlling low‐producer mutants in Bordetella pertussis cultures using antigenic modulation

“…Spontaneously occurring bvg – mutants grow faster and accumulate over generations [15]. To obtain a quantitative view of the growth rate difference between bvg – and bvg + cells, nine bvg – (non‐haemolytic) and nine bvg + (haemolytic) isolates derived from B. pertussis Tohama I (a strain used for vaccine production) were randomly selected and their growth rate was determined in high‐throughput growth assays in a chemically defined medium containing low amounts of niacin.…”

“…In the present study, cells were categorized as bvg + or bvg – on the sole basis of their haemolytic phenotype, which is linked to the production of AC [15]. Hence, avirulence was considered an all‐or‐nothing characteristic, neglecting: (i) the underlying genetic cause (mutations in different loci can result in a non‐haemolytic phenotype: AC structural genes, bvgAS locus, etc.…”

“…Avirulent bvg – mutants appear at a frequency of 10 –3 to 10 –6 per generation [2], and can be detected within as little as ∼20 generations [3]. They grow faster than bvg + cells and are more resistant to adverse conditions such as cryopreservation or nutrient limitation [15, 16].…”