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Antimicrobial activity of natural products from the flora of Northern Ontario, Canada.

Pharmaceutical biology (2015-02-24)
Janique Vandal, Mamdouh M Abou-Zaid, Garry Ferroni, Leo G Leduc
ABSTRACT

The number of multidrug resistant (MDR) microorganisms is increasing and the antimicrobial resistance expressed by these pathogens is generating a rising global health crisis. In fact, there are only a few antimicrobial agents left that can be used against MDR bacteria and fungi. In this study, the antimicrobial activities of selected natural products from the flora of Northern Ontario against selected microorganisms are reported. Plants were collected from Sault Ste. Marie, Ontario, Canada, and ethanol extracts were prepared using EtOH:H2O (1:1, v/v). Fungal cultures used in this study were Candida albicans ATCC 10231 and Schizosaccharomyces octosporus. Bacterial cultures employed included Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Mycobacterium phlei ATCC 11758, and Streptococcus lactis ATCC 19435. The microplate resazurin assay was used to screen for antimicrobial activity. Extracts of four plant species Chimaphila umbellata L. (Pyrolaceae), Betula papyrifera Marshall (Betulaceae), Rhus typhina L. (Anacardiaceae), and Fraxinus pennsylvanica Marshall (Oleaceae), and six compounds (gallic acid, ethyl gallate, caffeic acid, sinapic acid, gentisic acid, and chlorogenic acid) demonstrated antibacterial or antifungal activities with MICs ranging from 62.5 to 1000 µg/mL, respectively, for a chemical fraction of an extract from Betula papyrifera against the bacterium S. aureus. The present study has shown that certain plant extracts and select fractions and standard chemical compounds exhibit antimicrobial effects. Prince's Pine, Chimaphila umbellate, White Birch, Betula papyrifera, Staghorn Sumac, Rhus typhina, and Green Ash, Fraxinus pennsylvanica were the principal extracts exhibiting notable antibacterial and/or antifungal activities; while gallic acid, ethyl gallate, and caffeic acid demonstrated antibacterial activities and sinapic acid, gentisic acid, and chlorogenic acid demonstrated antifungal activities.

MATERIALS
Product Number
Brand
Product Description

USP
Vanillin Melting Point Standard, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Vanillic acid, ≥97%, FG
Supelco
Ethanol solution, certified reference material, 2000 μg/mL in methanol
Sigma-Aldrich
Vanillin, ReagentPlus®, 99%
Sigma-Aldrich
Vanillin, ≥97%, FCC, FG
Sigma-Aldrich
Methyl 3,4,5-trihydroxybenzoate, 98%
Sigma-Aldrich
Vanillin, natural, ≥97%, FCC, FG
Sigma-Aldrich
trans-3-Hydroxycinnamic acid, 99%
Sigma-Aldrich
Vanillic acid, 97%
Supelco
Sinapic acid, matrix substance for MALDI-MS, ≥99.0% (T)
Sigma-Aldrich
Ethanol Fixative 80% v/v, suitable for fixing solution (blood films)
Supelco
Ethanol standards 10% (v/v), 10 % (v/v) in H2O, analytical standard
Sigma-Aldrich
Glycerin, meets USP testing specifications
Sigma-Aldrich
Chlorogenic acid, ≥95% (titration)
Sigma-Aldrich
Sinapic acid, ≥98%, powder
Sigma-Aldrich
p-Coumaric acid, ≥98.0% (HPLC)
Sigma-Aldrich
Vanillic acid, purum, ≥97.0% (HPLC)
Sigma-Aldrich
Glycerol solution, 83.5-89.5% (T)
Sigma-Aldrich
Ethyl gallate, antioxidant, ≥96.0% (HPLC)
Supelco
Mettler-Toledo Calibration substance ME 18555, Benzoic acid, analytical standard, for the calibration of the thermosystem 900, traceable to primary standards (LGC)
Sigma-Aldrich
Vanillin, tested according to Ph. Eur.
Sigma-Aldrich
3-(Benzyldimethylammonio)propanesulfonate, BioXtra, ≥99.0% (HPCE)
Sigma-Aldrich
Ellagic acid, ≥95% (HPLC), powder, from tree bark
Supelco
Vanillin melting point standard, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Glycerin, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Dehydrated Alcohol, Pharmaceutical Secondary Standard; Certified Reference Material
Chlorogenic acid, primary reference standard
Supelco
Mettler-Toledo Calibration substance ME 51143093, Vanillin, traceable to primary standards (LGC)
Supelco
Ellagic acid, analytical standard
Supelco
Vanillic acid, certified reference material, TraceCERT®