• Title/Summary/Keyword: Disk-agar diffusion method

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Resistance Patterns of Frequently Applied Antimicrobials and Occurrence of Antibiotic Resistance Genes in Edwardsiella tarda Detected in Edwardsiellosis-Infected Tilapia Species of Fish Farms of Punjab in Pakistan

  • Kashif Manzoor;Fayyaz Rasool;Noor Khan;Khalid Mahmood Anjum;Shakeela Parveen
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.668-679
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    • 2023
  • Edwardsiella tarda is one of the most significant fish pathogens, causes edwardsiellosis in a variety of freshwater fish species, and its antibiotic resistance against multiple drugs has made it a health risk worldwide. In this study, we aimed to investigate the antibiotic resistance (ABR) genes of E. tarda and establish its antibiotic susceptibility. Thus, 540 fish (299 Oreochromis niloticus, 138 O. mossambicus, and 103 O. aureus) were collected randomly from twelve fish farms in three districts of Punjab in Pakistan. E. tarda was recovered from 147 fish showing symptoms of exophthalmia, hemorrhages, skin depigmentation, ascites, and bacteria-filled nodules in enlarged liver and kidney. Antimicrobial susceptibility testing proved chloramphenicol, ciprofloxacin, and streptomycin effective, but amoxicillin, erythromycin, and flumequine ineffective in controlling edwardsiellosis. Maximum occurrence of qnrA, blaTEM, and sul3 genes of E. tarda was detected in 45% in the liver, 58%, and 42% respectively in the intestine; 46.5%, 67.2%, and 55.9% respectively in O. niloticus; 24%, 36%, and 23% respectively in summer with respect to fish organs, species, and season, respectively. Motility, H2S, indole, methyl red, and glucose tests gave positive results. Overall, E. tarda infected 27.2% of fish, which ultimately caused 7.69% mortality. The Chi-squared test of independence showed a significant difference in the occurrence of ABR genes of E. tarda with respect to sampling sites. In conclusion, the misuse of antibacterial agents has led to the emergence of ABR genes in E. tarda, which in association with high temperatures cause multiple abnormalities in infected fish and ultimately resulting in massive mortality.

Antimicrobial resistance patterns of Listeria species and Staphylococcus aureus isolated from poultry carcasses in Korea (계육에서 분리한 Listeria species 와 Staphylococcus aureus의 항생제 내성패턴)

  • Hur, Jin;Kim, Jun Man;Kwon, Nam Hoon;Park, Kun Taek;Lim, Ji Youn;Jung, Woo Kyoung;Hong, Soon Keun;Park, Yong Ho
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.217-224
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    • 2004
  • This study was carried out to investigate the antibiotic resistance pattern of Listeria spp. and Staphylococcus aureus. A total of 17 (14.8%) L. monocytogenes, 13 (11.3%) L. innocua, 7 (7%) L. welshimeri, and 83 (72.2%) S. aureus were isolated from commercial poultry carcasses in Seoul and Kyonggi province during the period between 2001 and 2003. Antibiotic susceptibility test of all Listeria strains isolated was performed by the disk agar diffusion method. Antibiotics used in the study were as follows; Amikacin (An), Ampicillin (Am), Cephalothin (Cf), Chloramphenicol (C), Ciprofloxacin (Cip), Erythromycin (E), Gentamicin (Gm), Imipenem (Ipm), Kanamycin (K), Minocycline (Mi), Neomycin (N), Norfloxacin (Nor), Ofloxacin (Ofx), Penicillin (P), Streptomycin (S), Tetracycline (Te), Tobramycin (Nn), Trimethoprim (Tmp), Trimethoprim/Sulfamethoxazloe (Sxt), and Vancomycin (Va). The antibiotic resistance pattern of S. aureus isolates was performed by the disk agar diffusion method. For the latter program, antibiotics used to the study were as follows; Cf, C, Cip, Clindamycin (Cc), E, Gm, Ipm, Nafcillin (Nf), Oxacillin (Ox), P, Te, Sxt, and Va. Of the 17 L. monocytogenes isolates, 94.1% were resistant to Te, 88.2% to Mi, 11.8% to Nor, 11.8% to S, 5.9% to Cip, and 5.9% to C. Of 13 L. innocua, 53.8% were resistant to Te, 23.1% to Mi, 23.1% to S, 7.7% to Cip, and 7.7% to Nor. Of 7 L. welshimeri, 57.1% were resistant to Te, and 14.3% to Am. Of 83 S. aureus, 100% were resistant to Te, 86.7% to Gm, 34.9% to P, 15.7% to Cip, 12% to Cc, 9.6% to E. The multiple antibiotic resistance patterns of L. monocytogenes isolates were observed in Te Mi Cip (5.9%), Te Mi Nor (5.9%), Te Mi (76.5%), and Te Nor (5.9%). Multiple antibiotic resistance was also found in L. innocua isolates. Resistant to Te Mi S Cip Nor was 7.7%, Te Mi S (7.7%), Te Mi (7.7%), and was 7.7% to Te S. Antibiotic resistance patterns for S. aureus isolats were demonstrated to Te Gm P Cip Cc E (6.0%), Te Gm Cip Cc E (3.6%), Te Gm P Cc (1.2%), Te Gm P (15.6%), Te Gm Cip (2.4%), Te P Cip (2.4%), Te Gm Cc (1.2%), Te Gm (56.6%), Te P (9.6%), and to Te Cip (1.2%). The results of this study suggest a high incidence of Lsteria spp. and S. aureus on poultry carcasses. The contaminated poultry carcasses may be a potential vehicle for foodborne infections due to multiple antimicrobial resistant organisms.

Comparative Efficacy for Dressing Methods of Iodine on Infected Full Thickness Skin Wound in the Mouse (흰 쥐의 감염된 전층피부 결손에서 Iodine의 드레싱 방법에 따른 창상치유 효과의 비교)

  • Lee, Jong-Hoon;Kim, Yo-Han;Min, Kyung-Hee;Hong, Sung-Hee;Lee, Won-Mi;Chae, Jeong-Don;Jun, Jin-Hyun
    • Archives of Plastic Surgery
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    • v.37 no.3
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    • pp.213-219
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    • 2010
  • Purpose: Iodine has been used for the prevention or management of wound infection as a topical agent. Although iodine was widely used mainly by Betadine$^{(R)}$ and cadexomer iodine, there was no comparative study on the efficacies of dressing methods of iodine. And also it's wound healing effect was not yet clear. The purpose of this study is to compare antibacterial effects and wound healing effects associated with various dressing methods of iodine on infected full thickness skin defect in the mouse. Methods: One full thickness skin defects in the mice (n=60) were developed on the back and left open for twenty-four hours. Sixty mice were divided into four groups : group S (dressing with Betadine$^{(R)}$ soaking, n=15), group T (dressing with Betadine$^{(R)}$ topping, n=15), group I (dressing with Iodosorb$^{(R)}$, n=15), group G (control group, dressing with dry gauze, n=15). The size of the wound defects and the grades of wound healing were evaluated in 4, 7, 10 days, and antibacterial effect was evaluated with restricted zone in Mueller Hinton agar by disk diffusion method. Results: After the wound was left open for twenty-four hours, many Staphylococcus aureus were cultured. The wound defect size was decreased in order of Betadine$^{(R)}$ soaking, Iodosorb$^{(R)}$, Betadine$^{(R)}$ topping and gauze dressing group in all days, but difference among experimental groups was not statistically significant. The grade score of wound healing was increased in order of Betadine$^{(R)}$ soaking, Iodosorb$^{(R)}$, Betadine$^{(R)}$ topping and gauze dressing group, and the difference was statistically significant. Antibacterial effect for S. aureus was increased in order of Iodosorb$^{(R)}$, Betadine$^{(R)}$ soaking, Betadine$^{(R)}$ topping and gauze dressing group, and the difference was statistically significant. Conclusion: Selection of the effective dressing method of iodine for infected wounds remains a controversial decision. According to this study, Iodosorb$^{(R)}$ may be most effective method for antibacterial effect and Betadine$^{(R)}$ soaking may be most effective method for infected wound healing. However, further study is necessary to evaluate the clinical efficacy of dressing methods of iodine and to search for the mechanisms that explain their effects.

Characters of motile aeromonads isolated from imported ornamental fish (수입 관상어에서 분리한 motile aeromonads의 특성)

  • Jin, Se-yun;Ko, Chang-yong;Lee, Ye-ji;Jung, Yun-hee;Ju, Seong-cheol;Kim, Eunheui
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.89-96
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    • 2017
  • The majority of freshwater ornamental fish are imported and distributed domestically, causing high risk of exposure to exotic pathogens and drug resistant bacteria in Korea. Aeromonas hydrophila is known as a common species of fresh water bacteria and opportunistic fish pathogen, as well as a species causing zoonotic infection. In this study, we isolated motile aeromonads from various imported freshwater ornamental fish and studied the characters of the isolates. Imported freshwater ornamental fish were purchased on day 1 after the fish were deposited in the aquarium. Bacteria were isolated from the liver, kidney and spleen of fish using 0.5% NaCl containing tryptic soy agar medium. Bacteria were grouped on the basis of their morphological characteristics. The colonies with clear zone on starch-ampicillin agar (SA agar) were tentatively identified as Aeromonas spp. Two hundred and twenty-six strains, about 70% of total isolates were assumed to be Aeromonas spp. Nine isolates were further identified based on the result of the API 20E test and PCR using primers specific for A. hydrophila 16S rRNA gene. The isolates were identified as A. hydrophila and the API 20E test showed differences in trisodium citrate, D-sucrose, D-melibiose, amygdalin and L-arabinose availability between the nine isolates and standard A. hydrophila. The susceptibilities of the isolated bacteria to 10 antibacterial agents were confirmed by the disk diffusion method. Isolated strains were found to be resistant to amoxicillin and ampicillin and sensitive to florfenicol. However, 7 isolates showed multiple drug resistances to erythromycin, oxytetracycline, nalidixic acid etc. Pathogenicity of the isolates was determined by the artificial challenge test on goldfish (Carassius auratus). Three isolates caused 60 ~ 80% mortality in goldfish within 5 days after the initiation of challenge. These results indicate that multiple drug resistant, highly pathogenic and exotic A. hydrophila can spread to domestic aquarium and the preventive treatment of fish before sale is necessary.

Isolation of Salmonella from the layer chickens reacting in pullorum-typhoid agglutination test (추백리 혈청검사 양성 산란계로부터 Salmonella속균 분리)

  • 류재윤;전무형;장경수;손현수;곽학구;박경재;우용구
    • Korean Journal of Veterinary Service
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    • v.22 no.3
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    • pp.221-237
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    • 1999
  • To investigate the specificity of rapid slide agglutination test for pullorum-gallinarum diseases and to obtain a basic data for avian salmonellosis control, salmonella isolation was peformed for the layer chickens positively reacted in pullonlm-typhoid agglutination test. The biochemical, serological and antimicrobial properties of the isolates were examined. The results obtained through this study were summarized as follows; 1. Of 2,384 chickens tested by the agglutination test, 606 chickens (25.4%) were positive reactors. 154 of 606 reactors and 49 of the non-reacting chickens were investigated for salmonella isolation, resulting in isolation of 68 strains of salmonellae from 27 chickens. 2. By organs, the isolation frequency from liver, cecum, spleen, ovary and gall bladder showed 8.9% (18 strains), 8.9% (18 strains), 7.4% (15 strains), 4.4% (9 strains) and 3.9% (8 strains), respectively. 3. By culture medium the combination of selenite broth and MacConkey agar revealed the highest isolation rate and the enrichment culture by delayed secondary enrichment culture method was found the most effective for salmonella isolation. 4. The serotypes of 68 salmonella isolates were identified as 3 strains of S pullorum, 24 strains of S gallinarum, 15 strains of S typhimurium, 8 strains of S enteritidis, 7 strains of S paratyphi A, 5 strains of S typhimurium and 6 strains of the other salmonellae. 5. The serotypes of 8 salmonella strains isolated from 49 chickens non-reacting in pullorum-typhoid agglutination test were identified as 3 strains of S typhimurium and 5 strains of S infantis. 6. When 24 chickens of which 68 strains of salmonellae isolated were examined by microplate agglutination test, the average antibody titer for pullorum antigen was $2^{5.25}$. The chickens at antibody titer between $2^3$ and $2^5$ showed the higher frequency of isolation as compared with the chickens at the other titers. 7. When salmonella isolates were tested the antimicrobial drug sensitivity by disk diffusion method, S paratyphi A were highly sensitive by 100% to ATM and GM, S typhimurium, by 88% to AM, CIP, IMP and TN, S infantis, by 100% to AM, CRO, ENR and PIP, S enteritidis,by 100% to IMP and PIP, S pullorum, by 100% to ATM, CRO, ENR and PIP and S gallinarum, by 92% to CRO, CIP and PIP.

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A Study of Antibiotic Resistance of Enterococci Isolated from Recycling Water of Floor Fountains in Gwangju Area (광주지역내 바닥분수 재이용수에서 분리한 장구균의 항생제 내성 연구)

  • Kim, Yeon-Hee;Song, Hyeong-myeong;Choi, Yeong-Seop;Lee, Yoon-Kook;Park, Jin-Hwan;Kim, Sun-Jung;Kim, Ha-Ram;Kang, Yu-Mi;Bae, Seok-Jin;Cho, Yong-Gwan
    • Journal of Environmental Health Sciences
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    • v.45 no.3
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    • pp.222-230
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    • 2019
  • Objectives: This study aims to evaluate water quality in terms of microorganisms and identify the antibiotic resistance of Enterococci isolated from the recycling water in floor fountains at three parks and one reservoir in the Gwangju area. Methods: Water samples were analyzed for Enterococci using membrane Enterococcus indoxyl ${\beta}$ d glucoside agar (mEI) as described in USEPA Method 1600. The vancomycin-resistant Enterococci with VanA and VanB were identified by PCR. An examination of the antibiotic resistance of isolates against 14 antibiotics was performed by the disk diffusion method. Results: The drinking water quality criterion was exceeded for total colony counts in 68% of all recycling water samples. The average concentration of total califorms and fecal coliforms was 139,325 and 413 CFU/100 mL, respectively. VanA and VanB were not detected from the isolates. We found the antibiotic resistant Enterococci strains to be E. faecalis, E. faecium, E. durans, E. mundtii, E. hirae, and E. thailandicus. The isolates were resistant to Rifampin (50%), Erythromycin (25.8%), Tetracycline (10.2%), Nitrofurantoin (8.1%), Minocycline (3.1%), Erythromycin (1.2%), Penicillin (0.7%), Norfloxacin (0.5%), and Teicoplanin (0.5%) among the 14 antibiotics tested. Antibiotic resistance tests for Enterococci from the recycling water of floor fountains resulted in 30.2% showing resistance to two or more antibiotics. Conclusions: These results showed that the multi-antibiotic resistance of Enterococci, E. coli, and others should be investigated continuously in each environment field.

Adhesion control of Campylobacter jejuni in chicken skin using emulsifiers (유화제를 이용한 계육 표면에서 Campylobacter jejuni의 부착 제어)

  • Oh, Do Geon;Kim, Kwang Yup
    • Korean Journal of Food Science and Technology
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    • v.52 no.6
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    • pp.670-677
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    • 2020
  • To prevent contamination by Campylobacter jejuni during chicken carcass processing, the effect of emulsifiers on C. jejuni inoculated on chicken skin was investigated using confocal laser scanning microscopy. Among the 8 emulsifiers (SWA-10D, L-7D, M-7D, S-1670, L-1695, P-1670, polysorbate 20, polysorbate 80) tested for antimicrobial activity by the paper disk method, 4 emulsifiers (L-7D, L-1695, polysorbate 20, polysorbate 80) were screened further. Emulsifier L-1695 showed the largest clear zone at a concentration of 200 mg/mL. The 4 emulsifiers subjected to primary screening were screened for heat and pH stability. In the contact surface test, emulsifier L-1695 showed the lowest log CFU/㎠ value on both stainless steel and ceramic surfaces. When emulsifier L-1695 was applied via general and electrostatic spray methods, the number of C. jejuni entrapped inside chicken skin follicles was significantly reduced in both methods. In conclusion, the emulsifier L-1695 could be employed as a microbial detachment agent in the chicken carcass processing industry.

Comparison of Antibacterial Ointments for Antibacterial and Wound Healing Effect on Infected Full Thickness Skin Defect in the Mouse (흰 쥐의 감염된 전층피부결손에 대한 항균연고들의 항균효과 및 창상치유 효과의 비교)

  • Jeon, So-Yeon;Lee, Jong-Hoon;Min, Kyung-Hee;Hong, Sung-Hee;Lee, Won-Mi;Chae, Jeong-Don;Jun, Jin-Hyun
    • Archives of Plastic Surgery
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    • v.37 no.2
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    • pp.115-121
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    • 2010
  • Purpose: Many topical agents had been used for contaminated wound treatment. Although antimicrobial ointments were widely used as topical agents, their comparative antibacterial and wound healing effects were largely unreported. The purpose of this study was to compare antibacterial effects and wound healing effects of common topical ointments on infected full thickness skin defect in the mouse. Methods: One full thickness skin defects in the mice (n=60) were developed on the back and left open for twenty-four hours. Sixty mice were divided into four groups: group B (dressing with Bactroban$^{(R)}$, n=15), group I (dressing with Iodosorb$^{(R)}$, n=15), group T (dressing with Terramycin$^{(R)}$, n=15), group G (control group, dressing with dry gauze, n=15). The size of wound defects and the grades of wound healing were evaluated at 4, 7, 10 days, and antibacterial effect was evaluated with restricted zone in Mueller Hinton agar by disk diffusion method. After the wound was left open for twenty-four hours, many Staphylococcus aureus was cultured. The wound defect size was decreased in order of Bactroban$^{(R)}$ (B), Iodosorb$^{(R)}$ (I), Terramycin$^{(R)}$ and gauze dressing group in all days, but difference among experimental groups was not statistically significant. The grade score of wound healing was increased in order of Bactroban$^{(R)}$, Iodosorb$^{(R)}$, Terramycin$^{(R)}$ and gauze dressing group, and the difference was statistically significant. Antibacterial effect for was increased in order of Bactroban$^{(R)}$, Iodosorb$^{(R)}$, Terramycin$^{(R)}$ and gauze dressing group, and the difference was statistically significant. Conclusion: Topical antimicrobial ointments application was effective against wound infections by S. aureus. Bactroban$^{(R)}$ may be an optimal topical treatment for infected wounds according to this study. However, further study is necessary to evaluate the clinical efficacy of antimicrobial ointments and to search for the mechanisms that explain their effects.

Molecular Characteristics of Extended-Spectrum β-Lactamase Genes in Clinical Isolates of Escherichia coli and Klebsiella pneumoniae (임상검체에서 분리된 Escherichia coli와 Klebsiella pneumoniae의 Extended-Spectrum β-Lactamase 유전자형 및 분자유전학적 특성)

  • Chung, Kyung-Suk
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.1
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    • pp.26-33
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    • 2006
  • Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

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Anti-microbial Activity of Grapefruit Seed Extract and Processed Sulfur Solution against Human Skin Pathogens (피부질환 원인균에 대한 자몽종자추출물과 법제유황수의 항균 효과)

  • Ha, Yu-Mi;Lee, Bo-Bae;Bae, Hee-Jung;Je, Kyoung-Mo;Kim, Soon-Rae;Choi, Jae-Suk;Choi, In-Soon
    • Journal of Life Science
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    • v.19 no.1
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    • pp.94-100
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    • 2009
  • This study was carried out to examine the antimicrobial effects of grapefruit seed extract (GSE) and processed sulfur solution (PSS) against human skin pathogens: Malassezia furfur, M. restricta, Propionibacterium arnes, Trichophyton mentagrophytes and T. rubrum. The antimicrobial effects of GSE and PSS were tested by agar diffusion method and micro broth dilution method. As the results, the MIC values of GSE against M. furfur, M. restricta, P. acnes, T. mentagrophytes and T. rubrum were 3.91, 3.91, 0.004, 0.024, and $0.012{\mu}l/ml$, respectively. The MIC values of PSS were 0.03, 0.03, 0.156, 0.003, and $0.012{\mu}l/ml$, respectively. Antimicrobial activity of skin care emulsion products containing 0.5% GSE and 0.5% PSS against human skin pathogens were 5.2, 4.3, 8.0, 9.5 and 12.8 mm, respectively. Refractive index, pH, viscosity and color value of skin care emulsions containing GSE and PSS were measured. According to these results, it was concluded that the GSE and PSS were the promising sources of antibacterial agent which could be useful for skin and hair care products as well as for the alternative medicine development in treatment of certain types of skin ailments.