Aquaculture International

, Volume 27, Issue 5, pp 1503–1512 | Cite as

Retrospective diagnosis of archived marine fish experienced unexplained mortality reveals dual infections of Nocardia seriolae and Streptococcus iniae

  • Vuong Viet Nguyen
  • Channarong Rodkhum
  • Saengchan SenapinEmail author
  • Ha Thanh DongEmail author


Due to the limitation of accessible laboratory facilities for disease diagnosis at the time of the events, clinical samples of snubnose pompano (Trachinotus blochii) and orange-spotted grouper (Epinephelus coioides) were frozen preserved for retrospective investigation. This study employed a simple challenged test using crude homogenate prepared from the long-term frozen preserved tissues (~ 16 months) to infect Asian sea bass (Lates calcarifer) model. The experimental fish became sick and exhibited visceral white spots similar clinical signs previously observed in natural cases. Two morphologically different Gram-positive bacteria were concurrently recovered from the experimentally sick fish but not directly from the archived frozen tissue. Based on sequence homology of 16S rRNA, the bacterial strains were identified as Nocardia seriolae and Streptococcus iniae, respectively. Subsequent specific PCR assays confirmed co-infections of N. seriolae and S. iniae in the majority of naturally diseased fish preserved. Concurrent infection of N. seriolae and S. iniae is new to both pompano and grouper. This study also suggests that frozen preservation combined with challenged assay might be useful for retrospective diagnosis and recovery of infectious agents in under-equipped laboratories in Southeast Asia or elsewhere.


Concurrent infection Nocardia seriolae Streptococcus iniae PCR Retrospect 



The National Center for Genetic Engineering and Biotechnology (BIOTEC) under its human resource development program provided a research fellowship to VV Nguyen. The authors thank C. Techatanakitarnan for technical assistance.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and institutional guidelines for the care and use of animals were followed by the authors.


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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of Veterinary Microbiology, Faculty of Veterinary ScienceChulalongkorn UniversityBangkokThailand
  2. 2.National Broodstock Center for Mariculture SpeciesResearch Institute for Aquaculture No. 1Tu SonVietnam
  3. 3.Fish Infectious Diseases Research Unit (FIDs RU), Faculty of Veterinary ScienceChulalongkorn UniversityBangkokThailand
  4. 4.Fish Health Platform, Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of ScienceMahidol UniversityBangkokThailand
  5. 5.National Center for Genetic Engineering and Biotechnology (BIOTEC)National Science and Technology Development AgencyPathum ThaniThailand
  6. 6.Faculty of Science and TechnologySuan Sunandha Rajabhat UniversityBangkokThailand

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