title: Bordetella pertussis Isolates from Argentinean Whooping Cough Patients Display Enhanced Biofilm Formation Capacity Compared to Tohama I Reference Strain.
abstract: Pertussis is a highly contagious disease mainly caused by Bordetella pertussis. Despite the massive use of vaccines, since the 1950s the disease has become re-emergent in 2000 with a shift in incidence from infants to adolescents and adults. Clearly, the efficacy of current cellular or acellular vaccines, formulated from bacteria grown in stirred bioreactors is limited, presenting a challenge for future vaccine development. For gaining insights into the role of B. pertussis biofilm development for host colonization and persistence within the host, we examined the biofilm forming capacity of eight argentinean clinical isolates recovered from 2001 to 2007. All clinical isolates showed an enhanced potential for biofilm formation compared to the reference strain Tohama I. We further selected the clinical isolate B. pertussis 2723, exhibiting the highest biofilm biomass production, for quantitative proteomic profiling by means of two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry, which was accompanied by targeted transcriptional analysis. Results revealed an elevated expression of several virulence factors, including adhesins involved in biofilm development. In addition, we observed a higher expression of energy metabolism enzymes in the clinical isolate compared to the Tohama I strain. Furthermore, all clinical isolates carried a polymorphism in the bvgS gene. This mutation was associated to an increased sensitivity to modulation and a faster rate of adhesion to abiotic surfaces. Thus, the phenotypic biofilm characteristics shown by the clinical isolates might represent an important, hitherto underestimated, adaptive strategy for host colonization and long time persistence within the host.
authors.affiliation: CINDEFI-Centro Científico Tecnológico CONICET La Plata, Facultad de Ciencias Exactas, Universidad Nacional de La Plata Buenos Aires, Argentina.
authors.forename: Laura
authors.initials: L
authors.lastname: Arnal
authors.affiliation: Functional Microbiology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine Vienna Vienna, Austria.
authors.forename: Tom
authors.initials: T
authors.lastname: Grunert
authors.affiliation: CINDEFI-Centro Científico Tecnológico CONICET La Plata, Facultad de Ciencias Exactas, Universidad Nacional de La Plata Buenos Aires, Argentina.
authors.forename: Natalia
authors.initials: N
authors.lastname: Cattelan
authors.affiliation: Laboratory of Pediatric Infectious Diseases, Department of Pediatrics, Radboud University Medical Centre Nijmegen, Netherlands ; Laboratory of Medical Immunology, Department of Laboratory Medicine, Radboud University Medical Centre Nijmegen, Netherlands.
authors.forename: Daan
authors.initials: D
authors.lastname: de Gouw
authors.affiliation: CINDEFI-Centro Científico Tecnológico CONICET La Plata, Facultad de Ciencias Exactas, Universidad Nacional de La Plata Buenos Aires, Argentina.
authors.forename: María I
authors.initials: MI
authors.lastname: Villalba
authors.affiliation: CINDEFI-Centro Científico Tecnológico CONICET La Plata, Facultad de Ciencias Exactas, Universidad Nacional de La Plata Buenos Aires, Argentina ; Mikrobiologie, Institut for Biologie, Humboldt-Universitat zu Berlin Berlin, Germany.
authors.forename: Diego O
authors.initials: DO
authors.lastname: Serra
authors.affiliation: Netherlands Centre for Infectious Disease Control, National Institute for Public Health and the Environment Bilthoven, Netherlands.
authors.forename: Frits R
authors.initials: FR
authors.lastname: Mooi
authors.affiliation: Functional Microbiology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine Vienna Vienna, Austria.
authors.forename: Monika
authors.initials: M
authors.lastname: Ehling-Schulz
authors.affiliation: CINDEFI-Centro Científico Tecnológico CONICET La Plata, Facultad de Ciencias Exactas, Universidad Nacional de La Plata Buenos Aires, Argentina.
authors.forename: Osvaldo M
authors.initials: OM
authors.lastname: Yantorno
keywords: Bordetella pertussis; biofilm; clinical isolates; proteomic; real time PCR; whooping cough