kinds of antibiotics. The diversity of the drug resistance spectrum of 90 strains was also discussed. Results show that 90 V. harveyi strain

12 6 2 0 1 6 1 2 South China Fisheries Science Vol. 12, No. 6 Dec., 2016 doi: 10. 3969/ j. issn. 2095-0780. 2016. 06. 013 2012 2014 1, 2, 1 1,, 3 1,, 1, 1, 2, 1 ( 1.,, 510300; 2., 201306; 3., 572018) : ( Vibrio harveyi), K-B 90, Euclidean,, 90 26, 28. 9%, ( 3 ) 21, 35, 38. 9%, 80. 8%, 2012 2013 2014 1018 8 ;, 3 17132, 90 Group Group, 11124, 16. 9% 57. 1%100%, Group Group 5 i v,,, : ; ; ; : S 948 : A : 2095-0780 - ( 2016) 06-0099 - 09 Analysis of antibiotic resistance spectrum of Vibrio harveyi strains isolated from maricultured fish in the South China Sea during 2012 2014 JIANG Kui 1, 2, XU Liwen 1, SU Youlu 1, WANG Yu 3, GUO Zhixun 1, XU Haidong 1, GAO Fang 1, 2, FENG Juan 1 ( 1. Key Lab. of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture; South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China; 2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 210306, China; 3. Tropical Aquaculture Research and Development Center of South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Sanya 572018, China) Abstract: Vibrio harveyi, a main pathogen of maricultured fish, causes great economic loss in aquaculture industry. We tested the temporal and spatial distribution of drug resistance of 90 V. harveyi strains isolated from maricultured fish in the South China Sea by Kirbybauer diffusion method. Phylogenetic of drug resistance was analyzed by squared Euclidean distance using bacteriostatic rings diameter of : 2015-12-11; : 2016-02 -23 : ( 2015XH03 ) ; ( A201501B14 ) ; ( 31502210 ) ; ( 2014 B040404057 ) : ( 1990 - ),,, E-mail: jiangkui0514 @ 163. com : ( 1973 - ),,,, E-mail: jannyfeng@ 163. com

100 12 12 kinds of antibiotics. The diversity of the drug resistance spectrum of 90 strains was also discussed. Results show that 90 V. harveyi strains formed 26 antibiogram types and antibiogramabundance was 28. 9%. Thirty-five strains, occuping 38. 9% of total strains, had 21 multi-drug antibiogram types ( resistant to 3 antibiotics), which was 80. 8% of spectral richness. The antibiogram types show significant difference of 90 V. harveyi strains in years and regions: 10, 18 and 8 in 2012, 2013 and 2014 years, respectively, while 17, 13 and 2 kinds in Hainan, Guangdong and Guangxi Provinces correspondently. Phylogenetic and cluster analysis indicate that 90 strains separated to Group, Groupand others clusters. Eleven antibiogram types with abundance of 16. 9% in Group, and 12 antibiogram types with 57. 1% in Group, while four kinds with 100% in other clusters. Groupand Groupcould be divided into five subgroups of i v. The results indicate that V. harveyi strains display complex antibiogram types in Hainan, and the resistance spectrum of V. harveyi changes and expands as time goes on. Key words: Vibrio harveyi; antibiogram; cluster analysis; maricultured fish Guangdong and Guangxi Provinces, ( Vibrio harveyi) NOR /AKDOX /AMP/ PEN/GEN/ CIP [ 19 ],,, DNA G + C ( Penaeus monodon) 46% 48%, 0. 44 1. 38 m, 3% NaCl ( TET) ( FUR ), TCBS, O/129 [ 1-2 ] ( SMZ) [ 20],,, [ 3-5 ], 2012 2014 90,,, 12, [ 6-7],, 4, [ 8-1 0] : 1), ; 2), ; 3), ; 4), 1, [ 1 1] [ 12-13 ],, 1. 1 [ 1 4-15], 90, ( clinical and ( 1), ( laboratory standards institute),, ) 56, ( S) ( I) ( R) 3, ( Formio niger) ( Epinephelus fuscoguttatus) (, ) ( Trachinotus ova-, tus) ( Siganus guttatus) [ 16], ( T. blochii) ( Plectropomus leopardus) ( KF) ( E. awoara) ( Rachycentron canadum) ( CEF) ( CEP) ( FOX) - ( AZT) [ 17-18 ] ) 31, DOX /AMP/PEN/ GEN/ CIP/, ( CHL) 96. 7%, 80%, 56. 7%,, 2012 7 2014 5 ( Lutjanus sebae) ; ( ( Nibea coibor)

6 : 2012 2014 101 1 Fig. 1 2. 1 Sampling sites of V. harveyi A Z 26, ( N. diacanthus) 28. 9%, 1, AI ( Hapalogenys nitens) J 2, 3 ( Epinephelus sp. ) ;, 45, 3 50%, 11. 5% ; C 1. 2 1, 9, ( K-B ), 10%, 3. 8% ; BDGHVY [ 2 1] 12 Z 3, Y Z, ( ) :, 12, ( CEF, 5 g - 1 ) ( GEN, 10 13. 3%, 27% ; K g - 1 ) ( TET, 30 g - 1 ), 1 ; 14 ( CIP, 5 g - 1 ) ( NOR, 10 g 3, 23, - 1 ) ( CHL, 30 g - 1 ) 25. 6%, 53. 8% ( T/S, 23. 7 /1. 25 g - 1 ) ( RIF, 5 g - 1 ) ( FUR, 100 g - 1 ) 10188,, ( AMO, 20 g - 1 ) ( ERY, 15 g - 1 ) ( ENR, 10 g - 1 ) 2012 7 12 22, ATCC25922, 10, 38. 5%, [ 22-24 ] ( clinical and laboratory standards institute) 20, 1. 3,, 69. 2%, SIR, 39. 1%,, 1 8, 12 FUR / T/ S/ CEFAMOFUR / RIF 2014,, 1 5 22, 8, 3 30. 8%, 36. 4% :, : 177 5 ; ; : 1. 4 SPSS 19. 0,, [ 2 5], Euclidean, 2 90,, 2012 2013 2014 1713 2 45. 5%, K, Z 8 2013 46, 18, : 1 3 ( 2)

南 方 水 产 科 学 102 图 2 90 株 哈 维 弧 菌 耐 药 谱 及 其 聚 类 分 析 CEF. 头 孢 克 肟 ; GEN. 庆 大 霉 素 ; TET. 四 环 素 ; R IF. 利 福 平 ; FUR. 呋 Fig. 2 CEF. C efixime; GEN. RIF. R ifampi c in; Gentami ci n; TET. 唑 酮; 环 丙 沙 星 ; NOR. CIP. AMO. 阿 莫 西 林 ; ER Y. 红 霉 素 ; 诺氟 沙星 ; ENR. CHL. 恩诺 沙 星; 氯 霉 素 ; T / S. 复 方 新 诺 明 ; 图 3, 表 1, 表 3 同此 A ntibiogram and cluster analysis of 90 V. harveyi strains Tetrac yc li ne; C IP. FUR. Furazolidone; 第 12 卷 Ciprofl oxac in; AMO. A moxi ci lli n; ERY. NOR. Norfloxac i n; C HL. c hloroampheni c ol ; T / S. Trimethoprim / Sulfamethoxazole; Erythromyc in ; ENR. Enrofl oxac in; The s ame c ase i n Fi g. 3, Tab. 1 and Tab. 3.

6 : 2012 2014 103 Tab. 1 1 Antibiogram types of V. harveyi strains number of type strain antibiogram number of type strain antibiogram A 42 FUR/ AMO N 1 T/ S/ CEF/ RIF/ TET B 1 FUR/ NOR/ AMO O 1 FUR/ T/ S/ CEF/ RIF/ TET C 9 AMO P 1 FUR/ T/ S/ CEF/ RIF/ TET/ ERY / AMO D 1 FUR/ CEF/ AMO Q 1 FUR/ T/ S/ CEF/ RIF/ ERY / AMO E 1 FUR/ RIF/ ERY / AMO R 4 FUR/ T/ S/ CEF/ RIF/ AMO F 1 FUR/ GEN/ ERY / AMO S 1 T/ S/ CEF/ RIF/ TET/ AMO G 5 FUR/ RIF/ AMO T 4 FUR/ T/ S/ CEF/ AMO H 2 FUR/ ERY / AMO U 1 FUR/ T/ S/ CEF/ ERY / AMO I 1 ERY / AMO V 1 FUR/ GEN/ AMO J 2 RIF/ AMO W 1 FUR/ T/ S/ CEF/ TET/ ERY / AMO K 1 X 1 FUR/ NOR/ RIF/CIP L 2 FUR/ T/ S/ CEF/RIF/ CHL/ TET/ ERY / AM Y 1 FUR/ NOR/ RIF M 3 FUR/ T/ S/ CEF/RIF/ CHL/ TET/ AMO Z 1 T/ S/ CEF/ RIF 3 2. 3 2 Fig. 3 Comparison of antibiotic resistance of V. harveyi strains, Group Group ( 3) 2 T/ SCEFRIFCHL 90 12 TET, Group ( 3 ), Group 2 FURNORCIPENR 93. 7%, 82. 3% ; 22% 30% ; AMO Group Group 2 FURTETCIP ENR 10% ;, Group Group, 2. 2 2. 4 90, 12 3 ( 2),, 21, 35 i v 5 ; 2, 38. 9%, 80. 8%,, iii Group, ; 3, iiiivv Group 5 Group Group 4, 90 Group Group, 1112 4, 16. 9% 57. 1%100%,,, ivv 20 12 3 ( 4),

104 12 21. 4% 25. 8% 100% FUR / T/ S/CEF RIF/ TET/ AMO FUR /GEN/ ERY /AMO FUR /NOR /RIF/ CIP T/ S/ CEF/ RIF/TETRIF/ ERY /AMO FUR /T/S/ CEF/ TET/ ERY / AMO FUR /T/ S/ CEF/ AMO 2 Tab. 2 Antibiotic resistance patterns of different V. harveyi subgroups group subgroup /% number of number of type rate strain types /% type abundance ( / ) antibiogram ( type / number of strains) Group i 58 8 30. 7 13. 8 C /9, A /40, B/1, D/1, E/1, F/1, G/3, H /2 ii 7 5 19. 2 71. 4 K/1, A /1, J/2, I/1, G/2 Group iii 6 4 15. 4 66. 7 L/1, M/3, N/1, O /1, iv 8 5 19. 2 62. 5 L/1, P/1, Q/1, R /4, S/1 v 7 4 15. 4 57. 1 T /4, U/1, V /1, W/1 other 4 4 15. 4 100 X /1, Y /1, Z/1, A /1 3 ( S) ( I) ( R) Tab. 3 Comparison of antibiotic resistance from susceptible, intermediate and resistance of V. harveyi results of resistant group /% resistance FUR T/ S GEN CEF NOR RIF CHL TET CIP ENR ERY AMO S Group 1. 54 96. 92 58. 46 92. 31 100 66. 15 100 95. 38 95. 38 96. 92 15. 38 1. 54 Group 9. 52 4. 76 66. 67 0 100 14. 29 71. 43 38. 1 80. 95 85. 71 14. 29 4. 76 25 75 100 75 50 0 75 100 75 50 50 75 I Group 18. 46 3. 08 40 6. 15 0 20 0 3. 08 4. 62 3. 08 78. 46 0 Group 0 0 33. 33 0 0 19. 05 4. 76 14. 29 19. 05 14. 29 57. 14 4. 76 0 0 0 0 0 25 25 0 0 50 50 0 R Group 80 0 1. 54 1. 54 0 13. 85 0 1. 54 0 0 6. 15 96. 92 Group 90. 48 95. 24 0 100 0 66. 67 23. 81 47. 62 0 0 28. 57 90. 48 75 25 0 25 50 75 0 0 25 0 0 25 3 3. 1,,, 28. 9%, ( [ 26-27], 3 ) 21, 35, 38. 9%, 80. 8%, 54 8, 42, 77. 8% [ 28 ] 2012 2014 90, 12,

6 : 2012 2014 105 4 3 Tab. 4 Three or more than three kinds of antibiotic resistance patterns in Guangdong, Hainan, Guangxi Provinces Guangdong antibiogram type number of strains Hainan antibiogram type number of strains Guangxi antibiogram type number of strains B 1 D 1 T 2 E 1 F 1 W 1 G 2 G 3 M 2 H 2 N 1 L 2 O 1 M 1 P 1 Q 1 R 1 R 3 S 1 T 2 Y 1 U 1 V 1 X 1 Z 1 3. 2 21,,,,,, 2012 2013 2014 ( 2, 1), 3 13 2, 3 ; ( 30% ) ;,, ( 90% ) ;,, 5, 3 18. 2% A ( FUR / AMO) C( AMO) R 32. 6% 13. 6%, 2013 2 ; ( FUR / T/S/ CEF/ RIF/AMO ) M ( FUR / T/ S/CEF/, ; RIF/ CHL/ TET/ AMO) J( RIF/ AMO) 02006 97. 2%, 83. 3% [ 19], 77. 7%,, [ 29],,,,, 2007 2012 2007 2012, 12, [ 30], 26, 2012 13, 3 17 50. 0% [ 30 ],,, 2007 2012, 2012 2014

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