High-throughput quantitative method for assessing coaggregation among oral bacterial species

• DOI: 10.1111/lam.12622
• Publication Date: 2016-07-25T15:52:36Z
• Authors: E. Levin-Sparenberg; J.M. Shin; E.M. Hastings; M. Freeland; H. Segaloff; A.H. Rickard; B. Foxman
• Keywords: 0301 basic medicine; Mouth; Microscopy, Confocal; Streptococci; Streptococcus; Bacterial Adhesion; High-Throughput Screening Assays; 03 medical and health sciences; Microbiology and Immunology; Coaggregation; Biofilms; Actinomycetes; Health Sciences; Actinomyces; Biotechnology

This paper describes a high-throughput method that relies upon microplate reader to score coaggregation 60 min postmixing, and use of high-speed real-time imaging technology describe the rate over time. The results visual, microplate, FlowCam(™) aggregation scores for oral bacteria Streptococcus gordonii, oralis, Actinomyces oris, whose ability coaggregate are well characterized, compared. Following mixing all possible pairs, top fraction supernatant was added quantify cell-density. Pairs were also passed through flow cell within each pair. Results from both approaches correlated with corresponding visual microscopic observations. microplate-based assay enables screening, whereas -based validates quantifies extent autoaggregation occur. Together these assays open door future in-depth studies among large panels test strains.Coaggregation between bacterial species is integral multi-species biofilm development. Difficulties in rapidly reproducibly identifying quantifying have limited mechanistic studies. demonstrates two complementary quantitative methods screen coaggregation. first approach uses other device. rapid detection candidate coaggregating pairs strains simultaneously while controlling variation replicates. allows analysis rates size aggregates formed.