P. acnes project
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Transcript of P. acnes project
The effects of twelve commonly used herbal infusions on Propionibacterium acnes
04/04/2015The University of Edinburgh
Acne
Widely spread disease
Prevalence: 81%-95% boys 79%-82% girls 54% women 40% men
Propionibacterium acnes causes Acne by triggering inflammation in the skin.
Propionibacterium acnes
Preferential anaerobic
Aerotolerant
Diphtheroid
Gram-positive
Rod-shaped
Bruggemann (2010)
Acnes in Mass Media
Zu et al. (2010)
Project Aims
To determine, which of the commonly used herbal remedies are the most effective against the bacterium.
To test the most effective herbal infusions in a controlled manner.
To compare results and determine the most effective herbal infusion.
Project Methods
Making infusions
Disc diffusion test
Dilutions & growth rate test
1. Making Infusions
50 mL of water is poured over 5 g dried herbs.
Everything is heated in a microwave until water starts to boil.
The infusions are taken out of the microwave, cooled down for 10 minutes and then filtered.
2. Disc Diffusion Test
100 µL of P. acnes culture is spread over a blood agar plate
3 discs are impregnated with the prepared herbal infusion and placed on the blood agar.
The plate is incubated at 37 °C until growth can be seen. The diameters of the resulting areas of inhibition are measured.
By analysing the results, the most effective herbal infusions are selected for further testing.
0.5 mL of infusion
9.5 mL of broth
3. Dilutions & Growth Rate Test
1st experiment 2nd experiment Positive control
0.25 mL of infusion
9.75 mL of broth
10 mL of broth
3. Dilutions & Growth Rate Test
Incubation at 37 °C
Taking A600 measurements at regular time intervals.
Interpreting the results.
Disc diffusion test resultsSpecies Effective Diameter (mm)
Red Raspberry 18.3
Green Tea 23.3
Basil 10.0
Rosemary -
Bay Leaves -
Rosehip -
Thyme -
Camomile -
Ginger -
Dandelion -
Elder Flowers -
Lavender -
Control -
Green Tea
Raspberry Leaf
Basil
Disc diffusion test resultsAntibiotic Effective
Tetracycline (10µg)
Tetracycline (25µg)
Tetracycline (100µg)
Chloramphenicol (25µg)
Chloramphenicol (50µg)
Streptomycin (10µg)
Streptomycin (25µg)
Trimethoprim (1.25µg)
Trimethoprim (2.5µg)
Ampicillin (25µg)
Novobiocin (5µg)
Oxacillin (5µg)
Sulphamethoxazole (25µg)
Disc diffusion test resultsAntibiotic Effective
Nitrofurantoin (50µg)
Cephalexin (30µg)
Augmentin (30µg)
Ciprofloxacin (5µg)
Erythromycin (5µg)
Penicilin G (1 unit)
Clindamycin (2µg)
Gentamicin (10µg)
Fusidic acid (10µg)
Colistin sulphate (100µg)
Kanamycin (30µg)
Nalidixic acid (30µg)
Antibiotics
Dilutions & growth rate test results
Time (h) Green Tea (1) Green Tea (2) Raspberry leaves (1) Raspberry leaves (2) Basil (1) Basil (2)
0 0 0 0 0 0 0
21.083 0.503 0.543 0.506 0.585 0.384 0.111
27.917 0.34 0.339 0.453 0.483 0.488 0.534
44.833 3.476 0.514 0.698 0.724 0.506 0.476
52.083 0.864 1.156 0.769 0.763 0.584 0.534
69.083 1.102 0.525 0.914 0.909 0.567 0.582
76 2.19 0.658 0.933 1.101 0.581 0.647
92.866 2.487 0.499 0.883 0.932 0.551 0.721
Absorbance
1st experiment – Infusion : Broth ratio = 0.5 : 9.5
Dilutions & growth rate test results
Dilutions & growth rate test results
2nd experiment – Infusion : Broth ratio = 0.25 : 9.75
Time (h)Green Tea
(1)Green Tea (2) Raspberry leaves (1) Raspberry leaves (2) Basil (1) Basil (2)
0 0.072 0.025 0.048 0.072 0.017 0.017
22.17 0.001 0.245 0.769 0.744 0.523 1.596
29.42 0.113 0.119 1.58 1.694 0.786 0.618
46.33 0.085 0.093 1.65 1.657 0.658 0.628
70.08 0.081 0.096 1.438 1.69 0.62 0.699
94.08 0.469 0.416 1.782 1.952 0.813 1.19
100.92 2.308 1.744 1.638 1.606 0.762 0.746
Absorbance
Time (h) Positive (1) Positive (2)
0 0.015 0.016
22.17 0.977 0.99
29.42 1.127 1.368
46.33 1.116 1.226
70.08 1.336 1.083
94.08 1.78 2.032
100.92 2.002 1.854
Absorbance
Our Main Findings
Green Tea, Raspberry Leaf and Basil infusions seem to have antibacterial properties against Propionibacterium acnes.
At higher concentrations, Basil seems to be the most effective infusion, followed by Raspberry. Difficult to determine the effectiveness of Green Tea.
At lower concentrations, Green Tea seems to be the most effective infusion, followed by Basil. Raspberry Leaf infusion doesn’s seem to have any significant effect.
Green Tea
In the disc diffusion tests and low concentration growth rate tests Green Tea produced significant inhibition of P. acnes growth.
One possible active ingredient might be Polyphenon-60, as it is a compound found in Green Tea and it has displayed a therapeutic effect on acne. (Jung et al., 2012)
Raspberry Leaf
In the disc diffusion tests and low concentration growth rate tests Raspberry Leaf produced significant inhibition of P. acnes growth.
Previous research has demonstrated antimicrobial effects of Raspberry on other bacterial species, although a concrete active ingredient is currently unknown. (Furuhata et al., 2001; Ryan et al., 2001)
Basil
In the disc diffusion tests and low & high concentration growth rate tests Basil produced significant inhibition of P. acnes growth.
Previous research has demonstrated antimicrobial effects of Basil oils on P. acnes. It has been hypothesized that the active ingredients could be methyl chavicol and methyl eugenol. (Viyochet al., 2006)
References1. Aubin G.G, Portillo M.E, Trampuz A, Corvec S. Propionibacterium
acnes, an emerging pathogen: From acne to implant-infections, from phylotype to resistance. Médecine et Maladies Infectieuses; 2014, June; 44(6): 241–250.
2. Bruggemann H. 2010. Skin: Acne and Propionibacterium acnes Genomics. In: Timmis N.K. ed. Handbook of Hydrocarbon and Lipid Microbiology. London: Springer, pp. 3221.
3. Furuhata K, Dogasaki C, Hara M, Fukuyama M. Antibacterial activities of several herbs on Legionella pneumophila. Journal of Azabu University (Japan); 2001, March; 1-2:15-20
4. Goulden V, Stables G.I, Cunliffe W.J. Prevalence of facial acne in adults. Journal of the American Academy of Dermatology; 1999, October; 41(4): 577-780.
5. Hoeffler U. Enzymatic and hemolytic properties of Propionibacterium acnes and related bacteria. Journal of Clinical Microbiology; 1977, December; 6(6): 555-558.
References5. Jung M.K, Ha S, Son J, Song J.H, Houh Y, Cho E, Chun J.H, Yoon
S.R, Yang Y, Bang S.I, Kim M, Park H.J, Cho D. Polyphenon-60 displays a therapeutic effect on acne by suppression of TLR2 and IL-8 expression via down-regulating the ERK1/2 pathway. Arch Dermatol Res; 2012, June; 304:655-663
6. Perry A.L, Lambert P.A. Propionibacterium acnes. Letters in Applied Microbiology; 2006, February; 42(3): 185-188.
7. Purvis D, Robinson E, Merry S, Watson P. Acne, anxiety, depression and suicide in teenagers: A cross-sectional survey of New Zealand secondary school students. Journal of Paediatrics and Child Health; 2006, December; 42: 793–796.
8. Ryan T, Wilkinson J.M, Cavanagh H.M.A. Antibacterial activity of raspberry cordial in vitro. Research in Veterinary Science; 2001, December; 71(3):155-159
References9. Viyoch J, Pisutthanan N, Faikreua A, Nupangta K, Wangtorpol K,
Ngokkuen J. Evaluation of in vitro antimicrobial activity of Thai basil oils and their micro-emulsion formulas against Propionibacterium acnes. International Journal of Cosmetic Science; 2006; 28:125-133
10. Zu Y, Yu H, Liang L, Fu Y, Efferth T, Liu X, Wu N. Activities of ten essential oils towards Propionibacterium acnes and PC-3, A-549 and MCF-7 cancer cells. Molecules; 2010, April; 15(5): 3200-3210.
Acknowledgements
We would like to thank:
Edinburgh University Young Scientific Researchers Association (EUYSRA)
JCMB Biology Teaching Lab
Acknowledgements
The project team:
Paulius Eidukas
Agnietė Kampaitė
Raminta Kazlauskaitė
Tadas Andriuškevičius
Karolina Misevičiūtė
Thank you!