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264 วารสารการแพทย์แผนไทยและการแพทย์ ทางเลือก ปีที่ 18 ฉบับที่ 2 พฤษภาคม-สิงหาคม 2563

assay, respectively. tifungal activities respectively. Filter discs
impregnated with 10 mL of distilled water were
…. (2) used as a negative control. Solvent control disc

(ethanol) was also placed with the test, positive
where CPM sample is the radioactive count and negative control.

rate of the assay with enzyme and CPM The minimum inhibitory concentration
background is the same but without enzyme. (MIC) of the plant extracts was determined by
CPM total count is the count rate of 25 ml of using sterile 2 mL (96-well plates). Clindamy-

substrate plus 2 mL of buffer 1. Sildenafil was cin (0.1 mg/mL) and ampicillin (0.1 mg/mL)
a positive control. were used as controls for the S. aureus, Ps.

6. Antimicrobial screening aeruginosa and E. coli assays respectively.
Antimicrobial activity of T. trique- The deep-wells were incubated for 24 hours at
trum has been reported by folk medicine. 37 °C. The resulting turbidity was observed,

Bacterial growth inhibition was examined in and after 24 hours, MIC was determined to be
Staphylococcus aureus, Escherichia coli, Pseu- where growth was no longer visible by assess-
domonas aeruginosa, and Candida albicans. ment of turbidity by optical density readings

They are the representative of gram positive, at 600 nm with a Thermo Scientific Evolution
negative bacterium and fungus. The extracts 200 series UV-Vis Spectrophotometer. At least
were tested using 6 mm sterilized filter paper three replications were run for each assay.

discs. Discs were impregnated with 25 mL
(100 mg/mL concentration) of the T. trique- Results

trum extracts, allowed to dry and placed onto We examined the bioactivity of extracts
inoculated plates. Plates inoculated with E. of T. triquetrum prepared by different solvents.
coli, Ps. aeruginosa, S. aureus and C. albicans Then we examined those results to suggest

were incubated at 37 C for 24 hours, then which types of extracts may be appropriate
o
the diameters of the inhibition zones were to develop into health applications.

measured in millimeters. Each antimicrobial 1. Cytotoxicity testing
assay was performed in triplicate and mean First, we examined the cytotoxic ef-
values were reported. Standard antibiotics, fects of T. triquetrum extracts drawn out by

clindamycin (10 mg/ disc), ampicillin (10 mg/ different solvents on human fibroblasts. We
disc) and amphotericin-B (10 mg/ disc) served determined cytotoxic effect on fibroblast cells
as positive controls of antimicrobial and an- by using MTT assay. We measured the survival