Planta Med 2007; 73(12): 1275-1280
DOI: 10.1055/s-2007-981614
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

Investigation of Antibacterial Activity of Rosemary Essential Oil against Propionibacterium acnes with Atomic Force Microscopy

Yujie Fu1 , Yuangang Zu1 , Liyan Chen1 , Thomas Efferth2 , Huili Liang1 , Zhiguo Liu1 , Wei Liu1
  • 1Key Laboratory of Forest Plant Ecology, Ministry of Education, Northeast Forestry University, Harbin 150040, P. R. China
  • 2German Cancer Research Center, Heidelberg, Germany
Further Information

Publication History

Received: March 30, 2007 Revised: July 19, 2007

Accepted: July 26, 2007

Publication Date:
24 September 2007 (online)


In the present study, the antibacterial activity of rosemary (Rosmarinus officinalis L. Labiatae) essential oil against Propionibacterium acnes (P. acnes) was observed with atomic force microscopy (AFM). The MIC (minimal inhibitory concentration) value of rosemary essential oil against P.acnes was 0.56 mg/mL. Significant changes in morphology and size of P. acnes were observed by atomic force microscopy (AFM) in response to essential oil treatment. The essential oil first attached to the surface of P. acnes at low concentration, the width and height of the bacterial body became larger, whereas the length did not change considerably. With increasing concentration of the essential oil, the bacterial bodies were severely damaged. The length, width and height were all reduced, when the concentration was increased up to 64 × MIC, the length, width and height were reduced by 42.56 %, 92.00 % and 41.58 %, respectively. Furthermore, treated bacteria lost their native shape, the cell wall desquamated, and the cytoplasm leaked out of the bacterial body, finally leading to bacterial death. With the increasing time at MIC, the bacteria length was reduced at 8 h, the width and height gradually became smaller, the shape of the cell became distorted, and finally led to cell wall damage and bacterial death at 8 h. In conclusion, the AFM investigation of morphology and size of P. acnes treated with rosemary essential oil represents a powerful technique, which can generally be applied to reveal the biological changing mechanisms of bacteria induced by antibacterial agents at the nanometer level.


AFM: atomic force microscopy

BHI: brain heart infusion

MBC: minimal bactericidal concentration

MIC: minimal inhibitory concentration

P. acnes: Propionibacterium acnes


  • 1 Park J, Lee J, Jung E, Park Y, Kim K, Park B. et al . In vitro antibacterial and anti-inflammatory effects of honokiol and magnolol against Propionibacterium sp.  Eur J Pharmacol. 2004;  496 189-95.
  • 2 Cohmnawang M T, Surassmo S, Nukoolkarn V S, Gritsanapan W. Antimicrobial effects of Thai medicinal plants against acne-inducing bacteria.  J Ethnopharmacol. 2005;  101 330-3.
  • 3 Jappe U. Pathological mechanisms of acne with special emphasis on Propionibacterium acnes and related therapy.  Acta Derm Venerol. 2003;  83 241-8.
  • 4 Poulin Y. Practical approach to the hormonal treatment of acne.  J Cutan Med Surg. 2004;  4 16-21.
  • 5 Tan A W, Tan H H. Acne vulgaris: a review of antibiotic therapy.  Expert Opin Pharmacother. 2005;  6 409-18.
  • 6 Leyden J J. Antibiotic resistance in the topical treatment of acne vulgaris.  Cutis. 2004;  73 6-10.
  • 7 Yemisci A, Gorgulu A, Piskin S. Effects and side-effects of spironolactone therapy in women with acne.  J Eur Acad Dermatol Venerol. 2005;  19 163-6.
  • 8 Valero M, Salmeron M C. Antibacterial activity of 11 essential oil against Bacillus cereus in tyndallized carrot broth.  Int J Food Microbiol. 2003;  85 73-81.
  • 9 Butt H J, Wolff E K, Gould S A, Dixon Northern B, Peterson C M, Hansma P K. Imaging cells with the atomic force microscope.  J Struct Biol. 1990;  105 54-61.
  • 10 Binnig G, Quate C F, Gerber C. Atomic force microscope.  Phys Rev Lett. 1986;  56 930-3.
  • 11 Milhiet PE,Dosset P, Giocondi M C, Le Grimellec C. Atomic force microscopy: from cell imaging to molecular manipulation.  J Soc Biol. 2004;  198 169-74.
  • 12 Sullivan C J, Morrell J L, Allison D P, Doktycz M J. Mounting of Escherichia coli spheroplasts for AFM imaging.  Ultramicroscopy. 2005;  105 96-102.
  • 13 Bolshakova A V, Kiselyova O I, Filonov A S, Frolova O Y, Lyubchenko Y L, Yaminsky I V. Comparative studies of bacteria with an atomic force microscopy operating in different modes.  Ultramicroscopy. 2001;  86 121-8.
  • 14 Dufrene Y F. Application of atomic force microscopy to microbial surfaces: from reconstituted cell surface layers to living cells.  Micron. 2001;  32 153-65.
  • 15 Auerbach I D, Sorensen C, Hansma H G, Holden P A. Physical morphology and surface properties of unsaturated Pseudomonas putida biofilms.  J Bacteriol. 2000;  182 3809-15.
  • 16 Chada V G, Sanstad E A, Wang R, Driks A. Morphogenesis of Bacillus spore surfaces.  J Bacteriol. 2003;  185 6255-61.
  • 17 Yu J Q, Lei J C, Yu H D, Cai X, Zou G L. Chemical composition and antimicrobial activity of the essential oil of Scutellaria barbata .  Phytochemistry. 2004;  65 881-4.
  • 18 Sacchetti G, Maietti S, Muzzoli M, Scaglianti M, Manfredini S, Radice M. et al . Comparative evaluation of 11 essential oils of different origin as functional antioxidants, antiradicals and antimicrobials in foods.  Food Chem. 2005;  91 621-32.
  • 19 Wang Y, Song R, Li Y S, Shen J S. Understanding tapping-mode atomic force microscopy data on the surface of soft block copolymers.  Surf Sci. 2003;  530 136-48.
  • 20 Pang G K, Baba-Kishi K Z, Patel A. Topographic and phase-contrast imaging in atomic force microscopy.  Ultramicroscopy. 2000;  81 35-40.
  • 21 Dvorak J A. The application of atomic force microscopy to the study of living vertebrate cells in culture.  Methods. 2003;  29 86-96.
  • 22 Schaffer C, Messner P. The structure of secondary cell wall polymers: how Gram-positive bacteria stick their cell walls together.  Microbial Mol Biol Rev. 2005;  151 643-51.
  • 23 Navarre W W, Schneewind O. Surface proteins of gram-positive bacteria and mechanisms of their targeting to the cell wall envelope.  . 1999;  63 174-229.

Prof. Dr. Yuangang Zu

Key Laboratory of Forest Plant Ecology

Ministry of Education

Northeast Forestry University

Harbin 150040

People’s Republic of China

Phone: +86-451-8219-0535

Fax: +86-451-8210-2082