FACS Analysis of Bacterial Responses to Extracts of Vatica diospyroides Fruit Show Dose and Time Dependent Induction Patterns

Chuthapond MUSIMUN; Merika CHUYSONGMUANG; Patima PERMPOONPATTANA; Parinuch CHUMKAEW; Yaowaphan SONTIKUL; Nittaya UMMARAT; Theera SRISAWAT

Authors

Chuthapond MUSIMUN Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani 84000
Merika CHUYSONGMUANG Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani 84000
Patima PERMPOONPATTANA Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani 84000
Parinuch CHUMKAEW Department of Applied Chemistry, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani 84000
Yaowaphan SONTIKUL Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani 84000
Nittaya UMMARAT Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani 84000
Theera SRISAWAT Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani 84000 http://orcid.org/0000-0002-5467-1606

Keywords:

Antibacterial activity, bacteria, FACS analysis, response, Vatica diospyroides

Abstract

Antibacterial activity of Vatica diospyroides fruit extracts were screened against both Gram-positive and Gram-negative bacteria using disc diffusion method. Minimum Inhibitory Concentrations (MICs) were performed using a macro broth dilution assay. Bacteria were treated with the extracts at various dose levels (0.5, 1, and 2 MIC) and incubation times (3 and 6 h), and their responses were monitored by FACS analysis. The response patterns were analyzed in the counts of 4 subpopulations: viable cells (PI-, SSC-), membrane-damaged cells (PI+, SSC-), injured cells (PI-, SSC+), and dead cells (PI+, SSC+). The fraction of Bacillus subtilis responders to fruit cotyledon and pericarp extracts increased in a dose dependent manner, whereas Staphylococcus aureus treated with pericarp extract increased in a time dependent manner. Cells initially lost granularity (PI-, SSC+) and membrane integrity (PI+, SSC+). These results demonstrate that the FACS method can reveal the stages of bacterial responses to chemotherapeutic agents.

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