한수지 49(6), 838-844, 2016 Original Article Korean J Fish Aquat Sci 49(6),838-844,2016 Photobacterium damselae 의디스크확산법결과에대한 Epidemiological Cut-off Value 의결정 권문경 임윤진 1 김명석 2 서정수 김도형 1 * 국립수산과학원방역과, 1 부경대학교수산생명의학과, 2 국립수산과학원병리연구과 Epidemiological Cut-off Values Generated for Disc Diffusion Data from Photobacterium damselae Mun-Gyeong Kwon, Yun-Jin Lim 1, Myoung-Sug Kim 2, Jung-Soo Seo and Do-Hyung Kim 1 * Aquatic Life Disease Control Division, National Institute of Fisheries Science, Busan 46083, Korea 1 Department of Aquatic Life Medicine, Pukyong National University, Busan 48513, Korea 2 Pathology Division, National Institute of Fisheries Science, Busan 46083, Korea In this study, epidemiological cut-off values were estimated for 44 Photobacterium damselae isolates, since clinical breakpoints have not been established for this pathogenic bacterium. The susceptibility of the isolates to 10 antibiotics was evaluated using internationally standardized disc diffusion protocols. Normalized resistance interpretation was used to generate statistically valid epidemiological cut-off values for the susceptibility data. There were not enough strains exhibiting full sensitivity to ampicillin and amoxicillin to allow analysis of these antibiotics. Because there were only a marginally sufficient number of strains exhibiting full sensitivity to oxytetracycline, the cut-off value generated provided only a provisional estimate. The valid wild-type cut-off values were <13, 13, 9, 22, 25, 27, and 28 mm for gentamicin, cephalexin, erythromycin, oxolinic acid, flumequine, florfenicol, and sulfamethoxazole/ trimethoprim, respectively. The application of these cut-off values should greatly facilitate the rational selection of antibiotics for use in commercial fish farms. Key words: Photobacterium damselae, Antibiotic susceptibility, Disc diffusion, Epidemiological cut-off values, Antibiotic therapy 서론 (Wold Animal Health Organization, OIE) [Aquatic Animal Health Code (2016)] (http://www.oie.int/en/ international-standard-setting/aquatic-code/access-online/)., (Lim et al., 2016). CLSI (Clinical and Laboratory Standards Institute) CLSI M42-A (2006).,,, (interpretive criteria) Aeromonas salmonicida (CLIS, 2014). (interpretive criteria). clinical breakpoint., (pharmacokinetics; PK)/ (pharmacodynamics; PD) (susceptible) (resistant). http://dx.doi.org/10.5657/kfas.2016.0838 Korean J Fish Aquat Sci 49(6) 838-844, December 2016 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Received 5 October 2016; Revised 15 November 2016; Accepted 15 November 2016 *Corresponding author: Tel: +82. 51. 629. 5945 Fax: +82. 51. 629. 5938 E-mail address: dhkim@pknu.ac.kr These authors have contributed equally to this article. Copyright 2016 The Korean Society of Fisheries and Aquatic Science 838 pissn:0374-8111, eissn:2287-8815
Photobacterium damselae 의역학적 wild-type 기준값설정 839 PK/PD clinical breakpoint (Smith, 2012). Smith (2007). [European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2016a)] [minimum inhibitory concentration (MIC)] epidemiological cut-off values ( wild-type ; CO WT ). EUCAST wild-type (WT) ( ; ) non wild-type (NWT) ( ; ). clinical breakpoint., WT. CO WT clinical breakpoint. Kronvall (2003) normalized resistance interpretation (NRI) CO WT ; A. salmonicida (Ruane et al., 2007), Streptococcus phocae (Avendaño-Herreraa, 2011), Edwardsiella tarda Vibrio harveyi (Lim et al., 2016). Photobacterium damselae (Kwon et al., 2005) (Cho et al., 2013)., (Labella et al., 2011). P. damselae 50% Vibrio anguillarum V. harveyi (Kwon et al., 2005). P. damselae 44 NRI (Kronvall, 2003) wild-type non wild-type CO WT. 사용균주 재료및방법 P. damselae 2004 2014,,,,,,,,,,, 44 (Table 1). 1% NaCl tryptic soy agar (Difco, USA) 28, 18-24. 항생제감수성시험 P. damselae CLSI M42-A (2006) (disk diffusion test). CLSI M42-A (2006) EU- CAST (European committee on antimicrobial susceptibility testing, 2016b). 10 ; gentamicin (GEN), ampicillin (AMP), amoxicillin (AML), cephalexin (CFL), oxytetracycline (OTC), erythromycin (ERY), oxolinic acid (OA), flumequine (UB), florfenicol (FFC), trimethoprim/sulfamethoxazole (SXT) (Table 2)., 28 18-24 P. damselae 2 10 8 CFU/mL 1% NaCl Mueller-Hinton agar (MHA), 28 24-28 24. ( ). Quality control (QC) Escherichia coli ATCC 25922. Epidemiological cut-off value( 역학적 wild-type 기준값 ; CO WT ) 의설정 (Kronvall, 2003; Smith and Kronvall, 2014) NRI. NRI Bioscand AB, TA BY, Sweden (European Patent No. 1383913, US Patent No. 7,465,559). Wild type (normalized) zone probit. CO WT (Smith et al., 2007; Simth and Kronvall, 2014) 2.5. P. damselae. 억제대크기 결과 E. coli ATCC 25922 AMP,
840 권문경ㆍ임윤진ㆍ김명석ㆍ서정수ㆍ김도형 Table 1. Photobacterium damselae strains used in this study Isolate code Isolation source Isolation location Isolation year FP2235 Olive flounder (Paralichthys olivaceus) Pohang 2004 FP2237 Olive flounder (Paralichthys olivaceus) Pohang 2004 FP4137 Olive flounder (Paralichthys olivaceus) Jejudo 2004 FP2137 Olive flounder (Paralichthys olivaceus) Ulsan 2004 FP2244 Olive flounder (Paralichthys olivaceus) Pohang 2004 FP2261 Olive flounder (Paralichthys olivaceus) Jejudo 2005 FP2260 Olive flounder (Paralichthys olivaceus) Jejudo 2005 FP3092 Olive flounder (Paralichthys olivaceus) Ulsan 2005 FP3095 Olive flounder (Paralichthys olivaceus) Ulsan 2005 FP3099 Olive flounder (Paralichthys olivaceus) Ulsan 2005 FP3348 Olive flounder (Paralichthys olivaceus) Pohang 2007 FP8044 Olive flounder (Paralichthys olivaceus) Jejudo 2008 FP3737 Olive flounder (Paralichthys olivaceus) Pohang 2009 FP3738 Olive flounder (Paralichthys olivaceus) Pohang 2009 FP3739 Olive flounder (Paralichthys olivaceus) Pohang 2009 FP3741 Olive flounder (Paralichthys olivaceus) Pohang 2009 FP3755 Olive flounder (Paralichthys olivaceus) Ulsan 2009 FP4101 Olive flounder (Paralichthys olivaceus) Jejudo 2004 FP3047 Olive flounder (Paralichthys olivaceus) Ulsan 2004 FP3040 Olive flounder (Paralichthys olivaceus) Ulsan 2004 FP3348 Olive flounder (Paralichthys olivaceus) Pohang 2008 FP3525 Olive flounder (Paralichthys olivaceus) Pohang 2008 FP8482 Olive flounder (Paralichthys olivaceus) Wando 2010 FP8485 Olive flounder (Paralichthys olivaceus) Wando 2010 FP3392 Olive flounder (Paralichthys olivaceus) Pohang 2007 N8ATC-1 Olive flounder (Paralichthys olivaceus) Wild 2008 EP8-17 Olive flounder (Paralichthys olivaceus) Wando 2008 Dam 2-3-a Damselfish (Chromis motatus) Yeosu 2010 Gi 23-a Gizzard fish (Konosirus punctatus) Yeosu 2010 P.d4 mu Common mullet (Mugil cephalus) Tongyeong 2005 RSB Q3-2S Red sea bream (Pagrus major) Wando 2012 RSB R1-8cS Red sea bream (Pagrus major) Wando 2012 RB R2-2K Rock bream (Oplegnathus fasciatus) Wando 2012 SBe S3-3K Spottybelly greenling (Hexagrammos aframmus) Wando 2012 OF G2-1aS Olive flounder (Paralichthys olivaceus) Jejudo 2011 YT I4-1aS Yellow tail (Seriola quinqueradiata) Jejudo 2011 YT J1-3aS Yellow tail (Seriola quinqueradiata) Tongyeong 2011 PdRF-2nd Black rockfish (Sebastes schlegelii) Geomundo 2011 PdRF-3rd Black rockfish (Sebastes schlegelii) Geomundo 2011 PdSR Ib Shark ray (Rhina ancylostoma) Yeosu 2013 PdSR Ia Eagle ray (Myliobatis tobijei) Yeosu 2013 PdZe sh Zebra shark (Stegostoma fasciatum) Yeosu 2013 PdRF-02S Black rockfish (Sebastes schlegelii) Wild 2014 PdRF-04K Black rockfish (Sebastes schlegelii) Wild 2014
Photobacterium damselae 의역학적 wild-type 기준값설정 841 Table 2. Distribution of zone diameters (mm) for 44 isolates of Photobacterium damselae Zone diameters GEN AMP AML CFL OTC ERY OA UB FFC SXT 6 23 21 1 21 8 9 4 2 8 7 2 8 3 2 1 9 3 3 1 10 1 1 11 1 1 2 1 1 1 12 1 7 4 2 13 3 2 1 4 1 14 3 2 1 1 5 15 6 10 16 5 2 6 17 9 2 1 1 2 18 15 1 5 1 19 3 2 1 4 2 1 20 1 8 1 1 1 21 5 1 1 22 1 1 5 4 23 2 1 5 1 24 2 1 4 25 2 2 1 1 1 26 1 2 1 27 1 1 2 1 2 28 1 4 4 3 1 7 29 1 3 5 1 4 2 30 4 3 4 5 31 4 4 1 5 32 7 1 2 7 5 33 2 1 5 2 1 34 1 5 7 3 35 1 1 1 1 36 1 3 2 2 37 3 1 38 4 39 40 1 2 Abbreviations for antibiotic agent are as in the materials and methods. Areas highlighted in grey indicate the range of zones for isolates categorized as WT. GEN, gentamicin; AMP, ampicillin; AML, amoxicillin; CFL, cephalexin; OTC, oxytetracycline; ERY, erythromycin; OA, oxolinic acid; UB, flumequine; FFC, florfenicol; SXT, trimethoprim/sulfamethoxazole. GEN, ERY, SXT, OA, OTC, FFC CLSI M42-A (CLSI, 2006) quality control. P. damselae 44 Table 2. NRI 분석 10 data
842 권문경ㆍ임윤진ㆍ김명석ㆍ서정수ㆍ김도형 Table 3. Summary of results of NRI analysis Antibiotics Photobacterium damselae (n = 44) CO WT (mm) sd 1 (mm) WT 2 GEN 13 1.58 44 AMP AML Too few WT observations Too few WT observations CFL 13 3.14 37 OTC 28 1.28 20 ERY 9 1.81 36 OA 22 2.87 30 UB 25 3.67 33 FFC 27 2.78 37 SXT 28 2.16 27 Abbreviations for antibiotic agent are as in the materials and methods. 1 Standard deviation of the normalized distribution of WT observations. 2 Number of isolates categorized by the application of the relevant COwt value. GEN, gentamicin; AMP, ampicillin; AML, amoxicillin; CFL, cephalexin; OTC, oxytetracycline; ERY, erythromycin; OA, oxolinic acid; UB, flumequine; FFC, florfenicol; SXT, trimethoprim/sulfamethoxazole. sets AMP 9 NRI. AMP AML P. damselae 44 23 21 6 mm. NRI CO WT Table 3. 7, 6 5 NWT 3, 3 4. 고찰. P. damselae, ( non wild-type) ( wild-type). Cho et al. (2013) P. damselae CLSI (Enterobacteriaceae) clinical breakpoint. P. damselae, Photobacterium CLSI M100-S20 non- Enterobacteriaceae.. Thyssen and Ollevier (2001) P. damselae 145 Miller and Reimschuessel (2006) (eyeball technology) CO WT WT NWT. (Thyssen and Ollevier, 2001) P. damselae Chiu et al. (2013). (bimodal), ( ) NRI (Kronvall 2003). NRI (assumption) WT. Smith et al. (2012) Table 3 wild-type. AML 2.4 0.79 mm. AML 5.17 mm P. damselae AML. CLSI (2011) WT 30 WT MIC. NRI AMP AML P. damselae WT cut-off. NRI OTC WT 20 CLSI 30, OTC CO WT. P. damselae. Thyssen and Ollevier (2001) CO WT, AMP, AMX, OA, FFC, OTC, TE 6, 6, 24, 20, 24, 27 mm. AMP AMX 6 mm 23 21 non wild-type ( 80% ) Thyssen and Ollevier (2001) 10%. P. damselae. Thyssen and Ollevier (2001) ERY 6% WT, 44 82% WT. OTC, FFC, SXT P. damselae NWT (Thyssen and Ollevier, 2001) 10-30%,. P. damselae
Photobacterium damselae 의역학적 wild-type 기준값설정 843 Table 4. Antibiotic resistance patterns for Photobacterium damselae Type Antibiotic patterns No. of drugs Strains A1 AML, OTC, ERY, OA, UB, FFC, SXT 7 PdRF-2 nd (2011), PdRF-3 rd (2011) A2 AML, CFL, OTC, ERY, OA, UB, SXT 7 FP3755 (2009) A3 AML, OTC, OA, UB, FFC, SXT 6 FP2260 (2005), FP3040 (2004) A4 AML, OTC, ERY, OA, UB, FFC 6 FP4101 (2004) A5 AML, OTC, OA, UB, SXT 5 FP4137 (2004), FP3095 (2005) A6 AML, OTC, ERY, OA, UB 5 FP2261 (2005) A7 OTC, ERY, OA, UB, SXT 5 EP8-17 (2008) A8 AML, OTC, ERY, SXT 4 Gi 23-a (2010), RSB R1-8cS (2012) A9 AML, OTC, OA, SXT 4 FP8482 (2010) A10 AML, OTC, FFC, SXT 4 FP3739 (2009) R plasmid (Kawanishi et al., 2006; Nonaka et al., 2012),. 4 NWT 14, 7 NWT 2011 PdRF-2 PdRF-3 2009 FP 3755 P. damselae (Table 4). NWT. P. damselae. wild type WT NWT, Smith (2008) wild type WT NWT. WT 70% GEN, CFL, ERY, OA, UB, FFC P. damselae. P. damselae. SXT NWT 40%. P. damselae FFC SXT (Yasunaga and Yasumoto, 1988), Thyssen and Ollevier (2001) WT 99%, 15 NWT. P. damselae wild-type. Smith et al. (2007) EUCAST CLSI 3.. 사사 (R2016069). References Avendaño-Herreraa R, Molina A, Magariñosc B, Toranzoc AE and Smith P. 2011. Estimation of epidemiological cut-off values for disk diffusion susceptibility test data for Streptococcus phocae. Aquaculture 314, 44 48. http://dx.doi.org/10.1016/j. aquaculture.2011.01.049 Cho YA, Han HJ, Mun HE, Jung SH, Park MA and Kim JW. 2013. Characterization of Photobacterium damselae subsp. piscicida isolated from cultured starry flounder, Platichthys stellatus in Korea. J Fish Pathol 26, 77-88. http://dx.doi. org/10.7847/jfp.2013.26.2.077 Chiu TH, Kao LY and Chen ML. 2013. Antibiotic resistance and molecular typing of Photobacterium damselae subsp. damselae, isolated from seafood. J Appl Microbiol 114, 1184-1192. http://dx.doi.org/10.1111/jam.12104 CLSI. 2006. Methods for Antimicrobial Disk Susceptibility Testing of Bacteria Isolated From Aquatic Animals; Approved Guideline. CLSI document M42-A. Clinical and Laboratory Standards Institute, Wayne, Pennsylvania, U.S.A.
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