Mode of Action and Antibacterial Activity of Ethanolic Ant Plant Tuber Extract Inhibiting Growth of Staphylococcus aureus and Escherichia coli

Theera  SRISAWAT; Narueparn  SUKKASAM; Jirawadee  UPPALA; Chananchita  NILAKE; Kanokrat  KEAWCHAI; Amita  CHUJAN; Chuthapond  MUSIMUN; Parinuch CHUMKAEW; Patima  PERMPOONPATTANA

doi:10.48048/wjst.2021.9255

Authors

Theera SRISAWAT Faculty of Science and Industrial Technology, Prince of Songkla University, Suratthani Campus, Surat Thani 84000, Thailand http://orcid.org/0000-0002-5467-1606
Narueparn SUKKASAM Wittayanusorn Suratthani School, Muang, Surat Thani 84000, Thailand
Jirawadee UPPALA Wittayanusorn Suratthani School, Muang, Surat Thani 84000, Thailand
Chananchita NILAKE Wittayanusorn Suratthani School, Muang, Surat Thani 84000, Thailand
Kanokrat KEAWCHAI Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani Campus, Surat Thani 84000, Thailand
Amita CHUJAN Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani Campus, Surat Thani 84000, Thailand
Chuthapond MUSIMUN Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani Campus, Surat Thani 84000, Thailand https://orcid.org/0000-0002-8307-3760
Parinuch CHUMKAEW Department of Applied Chemistry, Prince of Songkla University, Surat Thani Campus, Surat Thani 84000, Thailand https://orcid.org/0000-0002-3924-2301
Patima PERMPOONPATTANA Department of Agricultural Science and Technology, Faculty of Science and Industrial Technology, Prince of Songkla University, Surat Thani Campus, Surat Thani 84000, Thailand https://orcid.org/0000-0002-0320-7184

DOI:

https://doi.org/10.48048/wjst.2021.9255

Keywords:

Antibacterial activity, Flow cytometry, Hydnophytum formicarum, Bacterial response, Resazurin assay

Abstract

The present study aimed to investigate the activity of Hydnophytum formicarum Jack (Ant Plant) tuber extract against Staphylococcus aureus and Escherichia coli and the mode of action of the extract on bacterial responses. The antibacterial activity was determined using agar well diffusion method. Resazurin-based 96-well microdilution method was used to determine the Minimal Inhibitory Concentrations (MIC) of the extract. Mode of action on bacterial death and response patterns to the extract were assessed by flow cytometry using membrane integrity and granularity profiles. The results, based on serial dilution and zones of inhibition against the bacteria, showed that 2,000 µg/well was potent against S. aureus (16.33±1.53 mm) and E. coli (14.33±0.58 mm). For S. aureus, the MIC concentration was 8,000 µg/mL, while 4,000 µg/mL concentration exhibited activity against E. coli. Flow cytometric profiles confirmed loss of intracellular components, followed by death, for both bacteria. Bacterial responses to the extract had consistent dose- and time-dependent behavior. This is the first mode of action study on bacterial responses to H. formicarum tuber extract. The results suggest that the extract of H. formicarum tuber could serve as a source of an initial active ingredient for developing effective medicines to battle infections caused by pathogens.

HIGHLIGHTS

Ant plant tuber extract had a strong antibacterial activity against S. aureus and E. coli
Intracellular component and bacterial membrane were the remarkable target for action of the extract
Dose- and Time-response model for bacterial response to the extract was achieved using Flow cytometry

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References

CL Ventola. The antibiotic resistance crisis: Part 1: Causes and threats. Pharm. Therapeut. 2015; 40, 277-83.

A Lee and J O'Rourke. Gastric bacteria other than Helicobacter pylori. Gastroenterol. Clin. North Am. 1993; 22, 21-42.

A Mailles and JP Stahl. Infectious encephalitis in France in 2007: A national prospective study. Clin. Infect. Dis. 2009; 49, 1838-47.

B Li and TJ Webster. Bacteria antibiotic resistance: New challenges and opportunities for implant-associated orthopaedic infections. J. Orthop. Res. 2018; 36, 22-32.

B Chakraborty, A Nath, H Saikia and M Sengupta. Bactericidal activity of selected medicinal plants against multidrug resistant bacterial strains from clinical isolates. Asian Pac. J. Trop. Med. 2014; 7, S435-S441.

O Pederson, A Gurib-Fakim, H Subratty and A Adsersen. Pharmacological properties of seven medicinal plants of the Rubiaceae from Mauritius. Pharm. Biol. 1999; 37, 202-7.

L Bore, T Honda and GW Gribble. Partial synthesis of 23-hydroxyursolic acid isolated from medicinal plants of the Rubiaceae family. Nat. Prod. Lett. 2002; 16, 273-6.

S Sen and R Chakraborty. Meyna spinosa Roxb.: An unexplored ethnomedicinal plant. Int. J. Green Pharm. 2017; 11, S332-S337.

S Prachayasittikul, P Buraparuangsang, A Worachartcheewan, C Isarankura-Na-Ayudhya, S Ruchirawat and V Prachayasittikul. Antimicrobial and antioxidative activities of bioactive constituents from Hydnophytum formicarum Jack. Molecules 2008; 13, 904-21.

O Prajuabjinda, S Panthong and A Itharat. Antimicrobial activity of Thai medicinal preparation of Khampramong temple used for cancer treatment and its plant components. J. Med. Assoc. Thai. 2012; 95, S159-S165.

T Hertiani and SUT Pratiwi,. Hydnophytum formicarum Jack ethanol extract modulates quorum sensing-controlled pathogenicity in Pseudomonas aeruginosa. Pak. J. Pharm. Sci. 2015; 28, 1691-7.

M Elshikh, S Ahmed, S Funston, P Dunlop, M McGaw, R Marchant and IM Banat. Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants. Biotechnol. Lett. 2016; 38, 1015-9.

C Musimun, M Chuysongmuang, P Permpoonpattana, P Chumkaew, Y Sontikul, N Ummarat and T Srisawat. FACS analysis of bacterial responses to extracts of Vatica diospyroides fruit show dose and time dependent induction patterns. Walailak J. Sci. Tech. 2017; 14, 883-91.

Y Andriani, H Mohamad, MNI Kassim, ND Rosnan, DF Syamsumir, J Saidin, TST Muhammad and H Amir. Evaluation on Hydnophytum formicarum tuber from Setiu wetland (Malaysia) and Muara Rupit (Indonesia) for antibacterial and antioxidant activities, and anti-cancer potency against MCF-7 and HeLa cells. J. Appl. Pharm. Sci. 2017; 7, 030-037.

TJ Silhavy, D Kahne and S Walker. The bacterial cell envelope. Cold Spring Harb. Perspect. Biol. 2010; 2, 1-16.

N Malanovic and K Lohner. Gram-positive bacterial cell envelopes: The impact on the activity of antimicrobial peptides. Biochim. Biophys. Acta 2016; 1858, 936-46.

AJ Drummond and RD Waigh. The development of microbiological methods for phytochemical screening. Recent Res. Dev. Phytochem. 2000; 4, 143-52.

SD Sarker, L Nahar and Y Kumarasamy. Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods 2007; 42, 321-4.

DA Hudman and NJ Sargentini. Resazurin-based assay for screening bacteria for radiation sensitivity. SpringerPlus 2013; 2, 55.

MR Brown, S Camezuli, RJ Davenport, E Petelenz-Kurdziel, L vreas and TP Curtis. Flow cytometric quantification of viruses in activated sludge. Water Res. 2015; 68, 414-22.

J Hong, W Guan, G Jin, H Zhao, X Jiang and J Dai. Mechanism of tachyplesin I injury to bacterial membranes and intracellular enzymes, determined by laser confocal scanning microscopy and flow cytometry. Microbiol. Res. 2015; 170, 69-77.

R Pagan and B Mackey. Relationship between membrane damage and cell death in pressure-treated Escherichia coli cells: Differences between exponential- and stationary-phase cells and variation among strains. Appl. Environ. Microbiol. 2000; 66, 2829-34.

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

DJ Dwyer, DM Camacho, MA Kohanski, JM Callura and JJ Collins. Antibiotic-induced bacterial cell death exhibits physiological and biochemical hallmarks of apoptosis. Mol. Cell. 2012; 46, 561-72.

T Senawong, S Misuna, S Khaopha, S Nuchadomrong, P Sawatsitang, C Phaosiri, A Surapaitoon and B Sripa. Histone deacetylase (HDAC) inhibitory and antiproliferative activities of phenolic-rich extracts derived from the rhizome of Hydnophytum formicarum Jack.: Sinapinic acid acts as HDAC inhibitor. BMC Complement. Altern. Med. 2013; 13, 232.

NS Abdullah, WYW Ahmad and NA Sabri. New compounds from Hydnophytum formicarum young tubers. Malaysian J. Anal. Sci. 2017; 21, 778-83.