101 DOI : 10.3831/KPI.2010.13.2.101 Received : 10. 05. 06 Accepted : 10. 05. 27 The Experimental Study on the Effect of Fel Ursi & Bovis Calculus Pharmacopuncture Solution in Bacterial Species which cause Keratitis Key Words: Fel Ursi, Bovis Calculus, Pharmacopuncture solution, anti-bacterial effect, keratitis. Na-young Han, Hyung-sik Seo Dept. of Ophthalmology, Otorhinolaryngology & Dermatology, Collego of Korean Medicine, Sangi University ABSTRACT Objectives : This experimental study was performed to investigate the effect of Fel Ursi & Bovis Calculus pharmacopuncture solution(fubcps) manufactured by using alcohol/water extraction method for identifying the use of it as eyedrops. Methods : FUBCPS was manufactured by using alcohol/water extraction method. Measure the size of inhibition zone and MIC(Minimum Inhibition Concentration) after administering FUBCPS on bacterial species, Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum and Candida albicans, which cause keratitis. Administering cravit(levofloxacin medicine) on bacterial species also performed to compare the anti-bacterial potency of this material, measured by using the size of inhibition zone. Results : After administering FUBCPS on bacterial species(staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, Candida albicans), there was no response to MIC and there was no anti-bacterial potency also. Conclusions : This study suggests that FUBCPS dose not have anti-bacterial effects on bacterial species which cause Keratitis. These study result recommends that we need to research more on herbal medicines of eye drop which have anti-bacterial effects on keratitis. I virus 65~90% Staphylococcus, Pseudomonas aeroginosa Fusarium Staphylococcus aureus Corresponding author : Hyungsik Seo. Department of Ophthalmology, Otorhinolaryngology and Dermatology, Korean Medicine Hospital, Sangji University, 283, Woosan-dong, Wonju-si, Kangwon-do, 220-955, South Korea. Tel: +82-33-741-9266 E-mail: aran99@sangji.ac.kr
102 Journal of Pharmacopuncture 13 2 2010 06 Staphylococcus epidermidis 6 Inhibition zone MIC II Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa Trypic Soy Agar Aspergillus niger Malt Extract Agar Fusarium oxysporum Potato Dextrose Agar Candida albicans Yeast Mold Agar 2 1 (1) (2) 90% alcohol alcohol 90% 90 10 magnetic stirrer 3 decompressor 90% alcohol alcohol 65 vacuum:15 5 (3) 80% alcohol 90% alcohol alcohol 80% 3 90% alcohol 200 alcohol 800 80% alcohol 1 1 (4) 70% alcohol 80% alcohol alcohol 70% 3 80%
103 alcohol 300 alcohol 700 (5) 70% alcohol 3 caramel alcohol 0.1 whatman, Germany (6) -30 PVTFD- 10A, ilshin lab, 100g 60~70g 2) (1) D.W 5 3 (2) (9) 100g 12g 3) (1) 50:50 0.006g/ 10cc vial 5cc chamber -50 20 4) Inhibition zone Minimum Inhibition Concentration, MIC (1) -80 37 24~48 20 Tryptic Soy broth 37 120rpm 18~24 OD Optical Density 1~2 10 8 25 0.2 Tween 80 10 1~2 10 8 (2) 100 filter disc cylinder cup 3 50 cylinder cup 150 sealing 37 1~2 25 3 1~3 Levofloxacin 5 Cravit III (2) ph 7.3 0.9 Nacl Na3PO4 Sodium phosphate 1. (MIC) 1) Staphylococcus aureus KCTC 1916
104 Journal of Pharmacopuncture 13 2 2010 06 Fig. 1 2) Staphylococcus epidermidis KCTC 1917 Fig. 2 3) Pseudomonas aeruginosa KCTC 2004 Fig. 3 4) Aspergillus niger KCTC 6906 Fig. 4 5) Fusarium oxysporum KCTC 16322 Fig. 5 6) Candida albicans KCTC 7965 Fig. 6 IV 5 Pseudomonas aeruginosa, Strep. pneumoniae, Moraxella, -hemolytic Streptococcus Fusarium Staphylococcus aureus S. aureus S. epidermidis Staphylococcus epidermidis 60 Pseudomonas 24
105 Fusarium spp. Aspergillus spp. Candida spp. Aspergillus Aspergillus Fusarium, Aspergillus, Candida Candida S.aureus S. pneumoniae Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa Aspergillus niger, Fusarium oxysporum, Candida albicans Cravit Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa 3 Cravit Staphylococcus aureus Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspe-
106 Journal of Pharmacopuncture 13 2 2010 06 rgillus niger, Fusarium oxysporum, Candida albicans 6 50 6 V Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, Candida albicans 1. Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, Candida albicans 50 6 VI 1. 2006:3,111,113. 2. 1999:75,81,82,97,100,101. 3. Park JH, Lee SB. Analysis on Inpatients With Infectious Keratitis: Causative Organisms, Clinical Aspects and Risk Factors. J Korean Ophthalmol Soc 2009;50(8):1152-66. 4. Jang YS, Hahn YH. Epidemiology of Staphylococcus epidermidis Keratitis. J Korean Ophthalmol Soc 2002;43(4):665-71. 5. 1985:104-8,429-30,432-34. 6. 3 1996:71,150,159,163. 7. Hahn YH, Hahn TW, Tchah HW, Choi SH, Choi KY, Kim KS, Wee WR, Kim JD, Kim HM, Chung JH, Lee HB, Kim JC, Jin KH, Yun YS, Myong YW, Chung SK, Joo CK, Kim MS, Ko MK, Kim EK, Lee JH, Kim HJ, Kim GB, Cho BJ, Kim WJ, Park WC, Lee JH. Epidemiology of Infectious Keratitis(II) : A Multi-center Study. J Korean Ophthalmol Soc. 2001;42(2):247-65. 8. Yang JH, Lee JH. The Antibacterial Activity of Domestic Topical Antibiotics -In Vitro-. J Korean Ophthalmol Soc. 1997;38(2):31-7. 9. Choi GD, Chae BY. The Effect of Coptidis Rhizoma, Fraxini, Jinpisan on Experimental Pseudomonas aeruginosa Keratitis. J Oriental Medical Surgery, Ophthalmology & Otolaryngology. 1999;12(1):18-35. 10. Smolin G, Thoft RA. The Cornea. 2nd e. Boston: Little Brown. 1987:205. 11. Lee JA, Kim HA, Lee YC. The literatual Study on Eye Drops. J Oriental Medical Surgery, Ophthalmology & Otolaryngology. 1993;12(2):155-70. 12. 1994:225. 13. 1982:477-8. 14. 2007:160,164,165,245, 257-9. 15. Ahn M, Jung YT, Han HJ. A Clinical Study on Infectious Corneal Ulcer. J Korean Ophthalmol Soc. 1996;37(9):126-31. 16. Hyon JY, Debasish Sinha, Terrence P. O' Brien, Lee JH, Wee WR. Expression of IRFs and
107 inos in the Mouse Cornea with Pseudomonas Aeruginosa Keratitis. J Korean Ophthalmol Soc. 2006;47(10):1654-61. 17. Hahn YH, Lee SJ, Hahn YW, Tchah HW, Wee WR, Kim KS. Epidemiology of Pseudomonas Keratitis: A Multi Center Study. J Korean Ophthalmol Soc. 1999;40(9):2411-22. 18. Davis SD: Pseudomonas:In Tasman W, Jaeger EA, eds. Duane's Foundations of Clinical Ophthalmology. Philadelphia. JB Lippincott,1990, :54. 19. Lee SJ, Lee JH. Three Cases of Pseudomonas Corneal Ulcer in Soft Contact Lens Wearer. J Korean Ophthalmol Soc. 1990;31(5):149-54. 20. Hahn YH, Byun KS. Efficacy of Fluconazole in The Treatment of Candida albicans Keratitis in Rabbits. J Korean Ophthalmol Soc. 1995;36(7):88-96. 21. Hahn YH, Lee DJ, Kim MS, Kim JD, Choi SH. Epidemiology of Fungal Keratitis in Korea : A Multi - center Study. J Korean Ophthalmol Soc. 2000;41(7):1499-508. 22. Kim YS, Song YS, Kim JC. Fungal Keratitis Caused by Chromomycetes. J Korean Ophthalmol Soc. 2003;44(3):755-9. 23. Hahn YH, Lee SJ, Hahn TW, Tchah HW, Kim JC. Antibiotic Susceptibilities of Ocular Isolates from Patients with Bacterial Ke ratitis :A Multi-center Study. J Korean Ophthalmol Soc. 1999;40(9):53-62. 24. Kim JC, Shyn KH, Hong NS. The Topical Effects of Antibiotics and Aprotinin in Experimental Pseudomonas Keratitis. J Korean Ophthalmol Soc. 1993;34(3):310-9. 25. Liu PV: Extracellular toxins of pseudomonas aeruginosa, J Infect Dis 1974; 9S4-99S:130 (spp). 26. Seo HS Lee YK, Roh SS. A Literaual Study on the External Treatment of Eye Diosease. The Journal of Oriental Medical Surgery, Ophthalmology & Otolaryngology. 1997;10(1):185-208. 27. Lee JA, Kim HA, Kee WC. Literatural Study on Eye-drop. The Journal of Oriental Medical Surgery, Ophthalmology & Otolaryngology. 1993;6(1):155-70. 28. Seo HS. The Experimental Study on Anti-bacterial Potency of Coptidis rhizoma extract on Staphylococcus aureus & Staphylococcus epidermidis. The Journal of Oriental Medical Surgery, Ophthalmology & Otolaryngology. 2006;19(2):71-76. 29. Seo HS. The Experimental Study on Anti-bacterial Potency of Coptidis rhizoma extract compare with quantity on Pseudomonas aeruginosa. Journal of Korean Pharmacopuncture institute. 2007;10(1):17-21. 30. Seo HS. The Experimental Study on Anti-bacterial Potency of Coptidis rhizoma extract compare with quantity on Staphylococcus epidermidis. Journal of Korean Pharmacopuncture institute. 2007;10(1):79-84 31. Seo HS. The Experimental Study on Anti-bacterial Potency of Coptidis rhizoma extract compare with quantity on Staphylococcus aureus. Journal of Korean Pharmacopuncture institute. 2007;10(3):47-52 32. Seo HS. The Experimental Study on the continuous Anti-bacterial Potency of Coptidis rhizoma extract for eye drop on Cultivation of Staphylococcus species(s. aureus, S. epidermidis). Journal of Korean Pharmacopuncture institute. 2007;10(3):71-76 33. Jung SY, Seo HS. The Experimental Study on Anti-bacterial Potency of Various Herbmedicine used for eye disease on Staphylococcus epidermidis Keratitis. The Journal of Oriental Medical Surgery, Ophthalmology & Otolaryngology. 2006;19(1):123-34. 34. Seo HS. The Experimental Study on Anti-bacterial Potency of Soyum Herbal-acupuncture on Staphylococcus aureus & Staphylococcus epidermidis. Journal of Korean Pharmacopuncture institute. 2006;9(2):73-77.
108 Journal of Pharmacopuncture 13 2 2010 06 35. Kang EK, Seo HS. The Experimental study of safety and efficacy in using Soyeom Pharmacopuncture solution as eyedrops. Journal of Korean Pharmacopuncture institute. 2009;12(1):21-33. 36. 3 2002:433-38. 37. Jeong YS, Lee GW, Gwon OH. In Vitro Activities of Levofloxacin and Other Commonly Used Antimicrobial Agents Aganist aerobic Bacteria. J Korean Infection Soc 1995;27(1):61-71. 38. Kang HJ. The Clinical Effect of CRAVIT (Levofloxacin) on Skin Infected Persons. Ewha Med J. 1996;19(3):359-64. Fig. 1. MIC of Fel Ursi & Bovis Calculus Pharmacopuncture Solutions and cravit on Staphylococcus aureus. Fel Ursi & Bovis Calculus Pharmacopuncture Solutions(3 /, 50 ) was not showed inhibition zone but cravit(levofloxacin, 5 ) was showed inhibition zone on Staphylococcus aureus. Fig. 2. MIC of Fel Ursi & Bovis Calculus Pharmacopuncture Solutions and cravit on Staphylococcus epidermidis. Fel Ursi & Bovis Calculus Pharmacopuncture Solutions(3 /, 50 ) was not showed inhibition zone but cravit(levofloxacin, 5 ) was showed inhibition zone on Staphylococcus epidermidis.
109 Fig. 3. MIC of Fel Ursi & Bovis Calculus Pharmacopuncture Solutions and cravit on Pseudomonas aeruginosa. Fel Ursi & Bovis Calculus Pharmacopuncture Solutions(3 /, 50 ) was not showed inhibition zone but cravit(levofloxacin, 5 ) was showed inhibition zone on Pseudomonas aeruginosa. Fig. 4. MIC of Fel Ursi & Bovis Calculus Pharmacopuncture Solutions and cravit on Aspergillus niger. Fel Ursi & Bovis Calculus Pharmacopuncture Solutions(3 /, 50 ) and cravit(levofloxacin, 5 ) were not showed inhibition zone on Aspergillus niger. Fig. 5. MIC of Fel Ursi & Bovis Calculus Pharmacopuncture Solutions and and cravit on Fusarium oxysporum. Fel Ursi & Bovis Calculus Pharmacopuncture Solutions(3 /, 50 ) and cravit(levofloxacin, 5 ) were not showed inhibition zone on Fusarium oxysporum.
110 Journal of Pharmacopuncture 13 2 2010 06 Fig. 6. MIC of Fel Ursi & Bovis Calculus Pharmacopuncture Solutions and cravit on Candida albicans. Fel Ursi & Bovis Calculus Pharmacopuncture Solutions(3 /, 50 ) and cravit(levofloxacin, 5 ) were not showed inhibition zone on Candida albicans.