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

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.


Keywords


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

Full Text:

PDF

References


VA Gant, G Warnes, I Phillips and GF Savidge. The application of flow cytometry to the study of bacterial responses to antibiotics. J. Med. Microb. 1993; 39, 147-54.

J Xu, F Zhou, BP Ji, RS Pei and N Xu. The antibacterial mechanism of carvacrol and thymol against Escherichia coli. J. Appl. Microbiol. 2008; 47, 174-9.

J Hong, W Guanc, G Jina, H Zhaoa, X Jianga and J Daia. Mechanism of tachyplesin I injury to bacterial membranes and intracellular enzymes, determined by laser confocal scanning microscopy and flow cytometry. J. Microbiol. Res. 2015; 170, 69-77.

F Silva, S Ferreira, JA Queiroz and FC Domingues. Coriander (Coriandrum sativum L.) essential oil: its antibacterial activity and mode of action evaluated by flow cytometry. J. Med. Microb. 2011; 60, 1479-86.

H Mathur, MC Rea, V Fallico, PD Cotter, C Hill and P Ross. Flow Cytometry as a tool to study the effects of bacteriocins on prokaryotic and eukaryotic cells. J. Mol. Diagn. 2016; S8, 013.

T Srisawat, P Chumkaew, W Heed-Chim, Y Sukpondma and K Kanokwiroon. Phytochemical screening and cytotoxicity of crude extracts of Vatica diospyroides Symington Type LS. Trop. J. Pharm. Res. 2013; 12, 71-6.

A Paparella, L Taccogna, I Aguzzi, CC Lopez, A Serio, F Marsilio and G Suzzi. Flow cytometric assessment of the antimicrobial activity of essential oils against Listeria monocytogenes. Food Control 2008; 19, 1174-82.

J Parekh and S Chanda. In vitro screening of antibacterial activity of aqueous and alcoholic extracts of various Indian plant species against selected pathogens from Enterobacteriaceae. Afr. J Microbiol. Res. 2007; 1, 92-9.

Y Jiang, N Wua, YJ Fu, W Wang, M Luo, CJ Zhao, YG Zua and XL Liu. Chemical composition and antimicrobial activity of the essential oil of rosemary. Environ. Toxicol. Pharm. 2011; 32, 63-8.

A Rezk, J Nolzen, H Schepker, DC Albach, K Brix and MS Ullrich. Phylogenetic spectrum and analysis of antibacterial activities of leaf extracts from plants of the genus Rhododendron. BMC Complement. Altern. Med. 2015; 15, 67.

MRA Largo, MG Nonato, EA Quinto and MA Tan. Antimicrobial polyphenol from the methanolic bark extract of Shorea contorta (Dipterocarpaceae). Asia-Pac J. Sci. Math. Eng. 2013; 1, 55-7.

K Lewis. Programmed death in bacteria. Microbiol. Mol. Biol. Rev. 2000; 64, 503-14.

KC Rice and KW Bayles. Molecular control of bacterial death and lysis. Microbiol. Mol. Biol. Rev. 2008; 72, 85-109.


Refbacks

  • There are currently no refbacks.




http://wjst.wu.ac.th/public/site/images/admin/image012_400

Online ISSN: 2228-835X

http://wjst.wu.ac.th

Last updated: 20 June 2019