Browsing by Author "Coenye, Tom"

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Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
(2021-07) Allkja, Jontana; van Charante, Frits; Aizawa, Juliana; Reigada, Ines; Guarch-Perez, Clara; Vazquez-Rodriguez, Jesus Augusto; Cos, Paul; Coenye, Tom; Fallarero, Adyary; Zaat, Sebastian A. J.; Felici, Antonio; Ferrari, Livia; Azevado, Nuno F.; Parker, Albert E.; Goeres, Darla M.
Microtiter plate methods are commonly used for biofilm assessment. However, results obtained with these methods have often been difficult to reproduce. Hence, it is important to obtain a better understanding of the repeatability and reproducibility of these methods. An interlaboratory study was performed in five different laboratories to evaluate the reproducibility and responsiveness of three methods to quantify Staphylococcus aureus biofilm formation in 96-well microtiter plates: crystal violet, resazurin, and plate counts. An inter-lab protocol was developed for the study. The protocol was separated into three steps: biofilm growth, biofilm challenge, biofilm assessment. For control experiments participants performed the growth and assessment steps only. For treatment experiments, all three steps were performed and the efficacy of sodium hypochlorite (NaOCl) in killing S. aureus biofilms was evaluated. In control experiments, on the log10-scale, the reproducibility SD (SR) was 0.44 for crystal violet, 0.53 for resazurin, and 0.92 for the plate counts. In the treatment experiments, plate counts had the best responsiveness to different levels of efficacy and also the best reproducibility with respect to responsiveness (Slope/SR = 1.02), making it the more reliable method to use in an antimicrobial efficacy test. This study showed that the microtiter plate is a versatile and easy-to-use biofilm reactor, which exhibits good repeatability and reproducibility for different types of assessment methods, as long as a suitable experimental design and statistical analysis is applied.
Minimum information guideline for spectrophotometric and fluorometric methods to assess biofilm formation in microplates
(2020-12) Allkja, Jontana; Bjarnsholt, Thomas; Coenye, Tom; Cos, Paul; Fallarero, Adyary; Harrison, Joe J.; Lopes, Susana P.; Oliver, Antonio; Pereira, Maria Olivia; Ramage, Gordon; Shirtliff, Mark E.; Stoodley, Paul; Webb, Jeremy S.; Zaat, Sebastian A. J.; Goeres, Darla M.; Azevedo, Nuno Filipe
The lack of reproducibility of published studies is one of the major issues facing the scientific community, and the field of biofilm microbiology has been no exception. One effective strategy against this multifaceted problem is the use of minimum information guidelines. This strategy provides a guide for authors and reviewers on the necessary information that a manuscript should include for the experiments in a study to be clearly interpreted and independently reproduced. As a result of several discussions between international groups working in the area of biofilms, we present a guideline for the spectrophotometric and fluorometric assessment of biofilm formation in microplates. This guideline has been divided into 5 main sections, each presenting a comprehensive set of recommendations. The intention of the minimum information guideline is to improve the quality of scientific communication that will augment interlaboratory reproducibility in biofilm microplate assays.
Minimum information guideline for spectrophotometric and ﬂuorometric methods to assess bioﬁlm formation in microplates
(Elsevier BV, 2020) Allkja, Jontana; Bjarnsholt, Thomas; Coenye, Tom; Cos, Paul; Fallarero, Adyary; Harrison, Joe J.; Lopes, Susana P.; Oliver, Antonio; Pereira, Maria Olivia; Ramage, Gordon; Shirtliff, Mark E.; Stoodley, Paul; Webb, Jeremy S.; Zaat, Sebastian A.J.; Goeres, Darla M.; Azevedo, Nuno Filipe
The lack of reproducibility of published studies is one of the major issues facing the scientific community, and the field of biofilm microbiology has been no exception. One effective strategy against this multifaceted problem is the use of minimum information guidelines. This strategy provides a guide for authors and reviewers on the necessary information that a manuscript should include for the experiments in a study to be clearly interpreted and independently reproduced. As a result of several discussions between international groups working in the area of biofilms, we present a guideline for the spectrophotometric and fluorometric assessment of biofilm formation in microplates. This guideline has been divided into 5 main sections, each presenting a comprehensive set of recommendations. The intention of the minimum information guideline is to improve the quality of scientific communication that will augment interlaboratory reproducibility in biofilm microplate assays.