, Volume 74, Issue 8, pp 1055–1062 | Cite as

Antibiotic susceptibility pattern of bacteria isolated from freshwater ornamental fish, guppy showing bacterial disease

  • Prasannan Geetha PreenaEmail author
  • Arathi Dharmaratnam
  • Nithianantham Sundar Raj
  • Thaliyil Veetil Arun Kumar
  • Shanmuganathan Arun Raja
  • Thangaraj Raja Swaminathan
Original Article


Antimicrobial resistance is one of the major threats faced in aquaculture systems. Hence the present study is mainly focused on to determine the antimicrobial susceptibility associated with pathogens derived from diseased freshwater ornamental guppy fishes. Around fifteen isolates were resolved from the infected fishes and subjected to antimicrobial susceptibility testing, phenotypic and genotypic characterization. Disc diffusion method was adopted for checking the antibiotic susceptibility using 17 antibiotic discs belonging to different classes. Dendrogram generated 5 clusters based on the biochemical tests and representative isolates were sequenced and identified as Aeromonas hydrophila, Aeromonas sobria, Pseudomonas putida, Acinetobacter soli and Kurthia gibsonii. The Shannon wiener diversity index of the resolved isolates was found to be 1.395 as determined by Primer-E software. Among the isolates, the majority of them was found to be Pseudomonas putida and exhibited higher antibiotic resistance towards antibiotics of 10 classes including third generation Cephalosporin and others showed the same against antibiotics of at least 5 classes tested. All of the recovered isolates possessed the MAR index of greater than 0.2, indicating the heavier dose of antibiotics in the farm. The detection of plasmid-mediated class I integron in Aeromonas hydrophila, Aeromonas sobria and Acinetobacter soli indicated the possibility of heavier dissemination of antimicrobial resistant genes among the ornamental fishes. Gentamycin and Ciprofloxacin were significantly effective against all the isolates and can be successfully applied in aquaculture. The occurrence of antibiotic resistance reminds the proper surveillance and continuous monitoring programmes in the fish farms and usage of other effective alternatives.


Antibiotic resistance Guppy fish Ornamental fish Aeromonas Pseudomonas Integron 



Antimicrobial Resistance


Antimicrobial Resistant Gene


Multiple Antibiotic Resistance


National Center for Biotechnology Information


Basic Local Alignment Search Tool



The corresponding author acknowledges Department of Science and Technology (DST) for the Fellowship under National Postdoctoral Fellowship Scheme with Grant number PDF/2017/000378 and National Bureau of Fish Genetic Resources for the support.

Compliance with ethical standards

Conflict of interest

The authors declare that we have no conflict of interest.


  1. Akinbowale OL, Peng H, Grant P, Barton MD (2007) Antibiotic and heavy metal resistance in motile aeromonads and pseudomonads from rainbow trout (Oncorhynchus mykiss) farms in Australia. Int J Antimicrob Agents 30:177–182. CrossRefPubMedGoogle Scholar
  2. Carnevia D, Letamendia M, Perretta A (2013) Pathogenic gram negative bacteria isolated from ornamental fish in Uruguay: characterization and antibiotic resistance. Bull Eur Assoc Fish Pathol 33:181–186Google Scholar
  3. Clarke KR, Gorley RN (2015) PRIMER v7: user manual/tutorial. PRIMER-E, Plymouth, p 296Google Scholar
  4. FAO (2003) The state of world fisheries and aquaculture 2002. FAO, Rome, ItalyGoogle Scholar
  5. Gao P, Mao D, Luo Y, Wang L, Xu B, Xu L (2012) Occurrence of sulfonamide and tetracycline-resistant bacteria and resistant genes in aquaculture environment. Water Res 46:2355–2364. CrossRefPubMedGoogle Scholar
  6. Hettiarachchi DC, Cheong CH (1994) Some characteristics of Aeromonas hydrophila and Vibrio species isolated from bacterial disease outbreaks in ornamental fish culture in Sri Lanka. J Natl Sci Counc Sri Lanka 22:261–269. CrossRefGoogle Scholar
  7. Jacobs L, Chenia HY (2007) Characterization of integrons and tetracycline resistance determinants in Aeromonas spp. isolated from south African aquaculture systems. Int J Food Microbiol 114:295–306. CrossRefPubMedGoogle Scholar
  8. Krumperman PH (1983) Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of fecal contamination of foods. Appl Environ Microbiol 46:165–170PubMedPubMedCentralGoogle Scholar
  9. Lee CR, Cho IH, Jeong BC, Lee SH (2013) Strategies to minimize antibiotic resistance. Int J Environ Res Public Health 10:4274–4304. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Lewbart GA (2001) Bacteria and ornamental fish. Semin Avian Exot Pet Med 10:48–56. CrossRefGoogle Scholar
  11. Luo Y, Mao D, Rysz M, Zhou Q, Zhang H, Xu L, Alvarez PJJ (2010) Trends in antibiotic resistance genes occurrence in the Haihe River, China. Environ Sci Technol 44:7220–7225. CrossRefPubMedGoogle Scholar
  12. Miller SA, Dyke DD, Polesk HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16:12–15Google Scholar
  13. Nabi N, Jabeen M, Hasnain A (2000) Recovery of multi drug resistant pseudomonads associated with an ulcerative condition in an air breathing murrel, Channa gachua. Asian Fish Sci 13:105–115Google Scholar
  14. Negrete RP, Romero JJ, Villegas LG, Vázquez SVC (2003) Presencia de Plasmidos en Pseudomonas aisladas de peces de ornato. Vet México 34:289–295Google Scholar
  15. O'Brien TF (2002) Emergence, spread and environmental effect of antimicrobial resistance: how use of an antimicrobial anywhere can increase resistance to any antimicrobial anywhere else. Clin Infect Dis 34(Suppl):3S78–3S84. CrossRefGoogle Scholar
  16. Okonko I, Donbraye-Emmanuel O, Ijandipe L, Ogun A, Adedeji A, Udeze A (2009) Antibiotics sensitivity and resistance patterns of uropathogens to nitrofurantoin and nalidixic acid in pregnant women with urinary tract infections in Ibadan, Nigeria. Middle- East. J Sci Res 4:105–109Google Scholar
  17. Osundiya OO, Oladele RO, Oduyebo OO (2013) Multiple antibiotic resistance (MAR) indices of Pseudomonas and Klebsiella species isolates in Lagos University teaching hospital. Afr J Cln Exper Microbiol 14(3):164–168. CrossRefGoogle Scholar
  18. Padrós F, Furones D (2004) Patología bacteriana en piscicultura. Boletin de la Sociedad Española de Microbiología 34:13–21Google Scholar
  19. Piotrowska M, Popowska M (2014) The prevalence of antibiotic resistance genes among Aeromonas species in aquatic environments. Ann Microbiol 64:921–934. CrossRefGoogle Scholar
  20. Schmidt AS, Bruun MS, Dalsgaard I, Larsen JL (2001) Characterization of class 1 integrons associated with R-plasmids in clinical Aeromonas salmonicida isolates from various geographical areas. J Antimicrob Chemother 47:735–743. CrossRefPubMedGoogle Scholar
  21. Sørum H (2006) Antimicrobial drug resistance in fish pathogens. In: Aarestrup FM (ed) Antimicrobial resistance in bacteria of animal origin. ASM Press, Washington, DC, pp 213–238Google Scholar
  22. Stalder T, Barraud O, Casellas M, Dagot C, Ploy MC (2012) Integron involvement in environmental spread of antibiotic resistance. Front Microbiol 3:119. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Torsvik V, Sorheim R, Gokseyr J (1996) Total bacterial diversity in soil and sediment communities-a review. J Ind Microbiol 17:170–l 78Google Scholar
  24. Verner-Jeffreys DW, Welch TJ, Schwarz T, Pond MJ, Woodward MJ, Haig SJ, Rimmer GS, Roberts E, Morrison V, Baker-Austin C (2009) High prevalence of multidrug-tolerant bacteria and associated antimicrobial resistance genes isolated from ornamental fish and their carriage water. PLoS One 4:e8388. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Walczak N, Puk K, Guz L (2017) Bacterial flora associated with diseased freshwater ornamental fish. J Vet Res 61:445–449. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Weir M, Rajic A, Dutil L, Cernicchiaro N, Uhland FC, Mercier B, Tusevljak N (2012) Zoonotic bacteria, antimicrobial use and antimicrobial resistance inornamental fish: a systematic review of the existing researchand survey of aquaculture-allied professionals. Epidemiol Infect 140:192–206. CrossRefPubMedGoogle Scholar
  27. Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Yonar ME, Karahan M, Kan N, Yonar S, Sağlam N (2010) A study of Acinetobacter sp. infection in some cultured rainbow trout (Oncorhynchus mykiss) in Kahramanmaras. J Fisheries Sciencescom 4:287–293Google Scholar

Copyright information

© Institute of Molecular Biology, Slovak Academy of Sciences 2019

Authors and Affiliations

  • Prasannan Geetha Preena
    • 1
    Email author
  • Arathi Dharmaratnam
    • 1
  • Nithianantham Sundar Raj
    • 1
  • Thaliyil Veetil Arun Kumar
    • 1
  • Shanmuganathan Arun Raja
    • 1
  • Thangaraj Raja Swaminathan
    • 1
  1. 1.Peninsular and Marine Fish Genetic Resources Centre of ICAR-NBFGRKochiIndia

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