TY - JOUR

T1 - A requirement for flow to enable the development of Ureaplasma parvum biofilms in vitro

AU - Rowlands, R. S.

AU - Kragh, K.

AU - Sahu, S.

AU - Maddocks, S. E.

AU - Bolhuis, A.

AU - Spiller, O. B.

AU - Beeton, M. L.

N1 - Publisher Copyright: © 2021 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.

PY - 2021

Y1 - 2021

N2 - Aims: To use a flow-based method to establish, quantify and visualize biofilms of Ureaplasma parvum. Methods and Results: Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour changing units (CCUs). Biofilms were established using the DTU flow-cell with a flow rate of 0·01 ml min−1 and compared to the static control. Titres of bacteria were determined by CCU and biofilm biomass was quantified by Syto9 staining and COMSTAT analysis. High-resolution images were obtained by scanning electron microscopy (SEM). Flow resulted in significantly more biofilm and higher cell titre (0·599 µm3/µm2 ± 0·152 and 4 × 108 CCU per ml, respectively) compared with static conditions (0·008 µm3/µm2 ± 0·010 and no recoverable cells, respectively). SEM revealed pleomorphic cells, with signs of budding and possible membrane vesicle formation. Conclusions: Flow is an essential requirement for the establishment of U. parvum biofilms. Significance and Impact of the Study: This is the first quantification of biofilm biomass formed by U. parvum. It is now possible to establish viable biofilms of U. parvum which will allow for future testing of antimicrobial agents and understanding of virulence-associated with adhesion.

AB - Aims: To use a flow-based method to establish, quantify and visualize biofilms of Ureaplasma parvum. Methods and Results: Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour changing units (CCUs). Biofilms were established using the DTU flow-cell with a flow rate of 0·01 ml min−1 and compared to the static control. Titres of bacteria were determined by CCU and biofilm biomass was quantified by Syto9 staining and COMSTAT analysis. High-resolution images were obtained by scanning electron microscopy (SEM). Flow resulted in significantly more biofilm and higher cell titre (0·599 µm3/µm2 ± 0·152 and 4 × 108 CCU per ml, respectively) compared with static conditions (0·008 µm3/µm2 ± 0·010 and no recoverable cells, respectively). SEM revealed pleomorphic cells, with signs of budding and possible membrane vesicle formation. Conclusions: Flow is an essential requirement for the establishment of U. parvum biofilms. Significance and Impact of the Study: This is the first quantification of biofilm biomass formed by U. parvum. It is now possible to establish viable biofilms of U. parvum which will allow for future testing of antimicrobial agents and understanding of virulence-associated with adhesion.

KW - biofilm

KW - flow cell

KW - quantification

KW - scanning electron microscopy

KW - Ureaplasma parvum

U2 - 10.1111/jam.15120

DO - 10.1111/jam.15120

M3 - Journal article

C2 - 33899996

AN - SCOPUS:85105328894

VL - 131

SP - 2579

EP - 2585

JO - Proceedings of the Society for Applied Bacteriology

JF - Proceedings of the Society for Applied Bacteriology

SN - 0266-8254

IS - 5

ER -