Abstract
The in vitro susceptibility of 63 isolates of Tenacibaculum maritimum from four fish farms to eight chemotherapeutic agents used for the treatment of bacterial diseases in fish were assessed. The results indicated that all strains were resistant to oxolinic acid and susceptible to amoxicillin, nitrofurantoin, florfenicol, oxytetracycline and trimethoprim-sulphamethoxazole. However, some isolates presented resistance to enrofloxacin and flumequine, ranging from 10 to 30%, and from 25 to 60%, respectively, depending on the farm sampled. These data were used in an attempt to predict whether the resistance to enrofloxacin was static or evolved during the time of sampling from 2003 to 2004. A relationship between the use of enrofloxacin and levels of resistance was detected in the studied farm, increasing significantly from no resistant isolates in 2003 to 44.8% resistant strains in 2004, the year in which this drug was commonly employed. This result was accompanied by a marked decline of about 29.2% of the inhibition zone sizes for the T. maritimum strains in comparison to the initial values (average 21.5 mm). Minimum inhibitory concentration (MIC) of enrofloxacin for 100 T. maritimum strains was determined by the microdilution method. Twenty isolates were resistant to enrofloxacin (> 256 μg ml−1), while the remaining strains showed a bimodal distribution, which ranged from 0.5 to 32 μg ml−1. Our interpretation of the enrofloxacin MIC data suggests that the breakpoint for T. maritimum should be 4 μg ml−1. However, similar studies in other laboratories are necessary to validate this breakpoint value.
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References
Alderman DJ, Hastings TS (1998) Antibiotic use in aquaculture: development of antibiotic resistance-potential for consumer health risks. Int J Food Sci Technol 33:139–155
Avendaño-Herrera R, Magariños B., López-Romalde S, Romalde JL, Toranzo AE (2004a) Phenotypic characterization and description of two major O-serotypes in Tenacibaculum maritimum strains isolated from marine fishes. Dis Aquat Org 58:1–8
Avendaño-Herrera R, Rodríguez J, Magariños B, Romalde JL, Toranzo AE (2004b) Intraspecific diversity of the marine fish pathogen Tenacibaculum maritimum as determined by randomly amplified polymorphic DNA-PCR. J Appl Microbiol 96:871–877
Avendaño-Herrera R, Núñez S, Magariños B, Toranzo AE (2004c) A non-destructive method for rapid detection of Tenacibaculum maritimum in farmed fish using nested PCR amplification. Bull Eur Assoc Fish Pathol 24:280–284
Avendaño-Herrera R, Irgang R, Núñez S, Romalde JL, Toranzo AE (2005) Recommendation of an appropriate medium for in vitro drug susceptibility testing of the fish pathogen Tenacibaculum maritimum. Antimicrob. Agents Chemother 49:82–87
Avendaño-Herrera R, Toranzo AE, Magariños B (2006) Tenacibaculosis infection in marine fish caused by Tenacibaculum maritimum: a review. Dis Aquat Org 71:255–266
Baquero F (1990) Resistance to quinolones in Gram-negative microorganisms: mechanisms and prevention.Eur Urol 17:3–12
Baxa DV, Kawai K, Kusuda R (1986) Characteristics of gliding bacteria isolated from diseased cultured flounder, Paralichthys olivaceous. Fish Pathol 21:251–258
Bernardet JF, Kerouault B, Michel C (1994) Comparative study on Flexibacter maritimus strains isolated from farmed sea bass (Dicentrarchus labrax) in France. Fish Pathol 29:105–111
Bowser PR, Wooster GA, Hsu HM (1994) Laboratory efficacy of enrofloxacin for the control of Aeromonas salmonicida infection in rainbow trout. J Aquat Anim Health 6:288–291
Bruun MS, Schmidt AS, Madsen L, Dalsgaard I (2000) Antimicrobial resistance patterns in Danish isolates of Flavobacterium psychrophilum. Aquaculture 187:201–212
Burka JF, Hammell KL, Horsberg TE (1997) Drugs in salmonid aquaculture-a review. J Vet Pharmacol Ther 20:333–349
Cepeda C, Santos Y (2002) First isolation of Flexibacter maritimus from farmed Senegalese sole (Solea senegalensis, Kaup) in Spain. Bull Eur Assoc Fish Pathol 22:388–391
Dalsgaard I, Bjerregaard J (1991) Enrofloxacin as an antibiotic in fish. Acta Vet Scand, Suppl 87:300–302
della Rocca G, Di Salvo A, Malvisi J, Sello M (2004) The disposition of enrofloxacin in seabream (Sparus aurata L.) after single intravenous injection or from medicated feed administration. Aquaculture 232:53–62
Devesa S, Barja JL, Toranzo AE (1989) Ulcerative skin and fin lesions in reared turbot, Scophthalmus maximus (L). J Fish Dis 12:323–333
Handlinger J, Soltani M, Percival S (1997) The pathology of Flexibacter maritimus in aquaculture species in Tasmania, Australia. J Fish Dis 20:159–168
Hansen JG, Sørheim R (1991) Improved method for phenotypical characterization of marine bacteria. J Microbiol Methods 13:231–241
Holt JG, Kieg NR, Sneath P, Staley JT, Williams ST (1994) Bergey’s Manual of Determinative Bacteriology, 9th edn. Williams and Wilkins, Maryland, USA, pp. 753
Hsu HM, Wooster GA, Bowser PR (1994) Efficacy of enrofloxacin for the treatment of salmonids with bacterial kidney disease, caused by Renibacterium salmoninarum. J Aquat Anim Health 6:220–223
Intorre L, Cecchini S, Bertini S, Cognetti-Varriale AM, Soldani G, Mengozzi G (2000) Pharmacokinetics of enrofloxacin in the seabass (Dicentrarchus labrax). Aquaculture 182:49–59
McVicar AH, White PG (1979) Fin and skin necrosis of cultivated Dover sole, Solea solea (L.). J Fish Dis 2:557–562
Miller RA, Walker RD, Baya A, Clemens K, Coles M, Hawke JP, Henricson BE, Hsu HM, Mathers JJ, Oaks JL, Papapetropoulou M, Reimschuessel R (2003) Antimicrobial susceptibility testing of aquatic bacteria: quality control disk diffusion ranges for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 at 22 and 28 °C. J Clin Microbiol 41:4318–4323
National Committee for Clinical Laboratory Standards. (2002) Performance standard for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals. Approved standard M31-A2. NCCLS, Wayne, Pennsylvania USA
National Committee for Clinical Laboratory Standards. (2003) Methods for Antimicrobial Disk Susceptibility Testing of Bacteria Isolated from Aquatic Animals; A Report. NCCLS document M42-R [ISBN 1–56238-501-1]. NCCLS, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA
Pazos F, Santos Y, Núñez S, Toranzo AE (1993) Increasing occurrence of Flexibacter maritimus in the marine aquaculture of Spain. FHS/AFS Newsletter 21:1–2
Pazos F, Santos Y, Macias AR, Nuñez S, Toranzo AE (1996) Evaluation of media for the successful culture of Flexibacter maritimus. J Fish Dis 19:193–197
Pursell L, Samuelsen OB, Smith P (1995) Reduction in the in-vitro activity of flumequine against Aeromona salmonicida in the presence of the concentrations of Mg2+ and Ca2+ ions found in sea water. Aquaculture 135:245–255
Santos Y, Pazos F, and Barja JL (1999) Flexibacter maritimus, causal agent of flexibacteriosis in marine fish. In: “ICES Identification Leaflets for Diseases and Parasites of Fish and Shellfish”. No. 55. International Council for the Exploration of the Sea. (ICES) (eds.). Denmark
Smith P, Hiney MP, Samuelsen OB (1994) Bacterial resistance to antimicrobial agents used in fish farming: a critical evaluation of method and meaning. Ann Rev Fish Dis 4:273–313
Sokal R, Rohlf J (1980) Introducción a la bioestadística. De. Reverte S.A., Barcelona
Soltani M, Shanker S, Munday BL (1995) Chemotherapy of Cytophaga/Flexibacter-like bacteria (CFLB) infections in fish: studies validating clinical efficacies of selected antimicrobials. J Fish Dis 18:555–565
Stoffregen DA, Chako AJ, Backman S, Babish JG (1993) Successful therapy of furunculosis in Atlantic salmon, Salmo salar L., using the fluoroquinolone antimicrobial agent enrofloxacin. J Fish Dis 16:219–228
Suzuki M, Nakagawa Y, Harayama S, Yamamoto S (2001) Phylogenetic analysis and taxonomic study of marine Cytophaga-like bacteria: proposal for Tenacibaculum gen. nov. with Tenacibaculum maritimum comb. nov. and Tenacibaculum ovolyticum comb. nov., and description of Tenacibaculum mesophilum sp. nov. and Tenacibaculum amylolyticum sp. nov. Int. J Syst Evol Microbiol 51:1639–1652
Toranzo AE, Novoa B, Romalde JL, Núñez S, Devesa S, Mariño E, Silva R, Martínez E, Figueras A, Barja JL (1993) Microflora associated with healthy and diseased turbot (Scophthalmus maximus) from three farms in northwest Spain. Aquaculture 114:189–202
Toranzo AE, Romalde JL, Dopazo CP, Magariños B, Barja JL (2004) Disease trends in the primary marine fish species cultured in Spain: A 20-year study. World Aquac 35:35–38
Toranzo AE, Magariños B, Romalde JL (2005) A review of the main bacterial fish diseases in mariculture systems. Aquaculture 246:37–61
Toyama T, Kita-Tsukamoto K, Wakabayashi H (1996) Identification of Flexibacter maritimus, Flavobacterium branchiophilum and Cytophaga columnaris by PCR targeted 16S Ribosomal DNA. Fish Pathol 31:25–31
Tsoumas A, Alderman DJ, Rodgers CJ (1989) Aeromonas salmonicida: development of resistance to 4-quinolone antimicrobials. J Fish Dis 12:493–507
Williams PJ, Courternay SC, Vardy C (1997) Use of enrofloxacin to control atypical Aeromonas salmonicida in Atlantic Tomcod. J Aquat Anim Health 9:216–222
Acknowledgements
This work was supported in part by grant AGL2004-07037 from the Ministerio de Ciencia y Tecnología (Spain). We would like to thank R. Irgang for her help in the MIC determination. R. Avendaño-Herrera also thanks Programs CONICYT-BID, Chile for a research fellowship.
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Avendaño-Herrera, R., Núñez, S., Barja, J.L. et al. Evolution of drug resistance and minimum inhibitory concentration to enrofloxacin in Tenacibaculum maritimum strains isolated in fish farms. Aquacult Int 16, 1–11 (2008). https://doi.org/10.1007/s10499-007-9117-y
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DOI: https://doi.org/10.1007/s10499-007-9117-y