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Bioencapsulation and Colonization Characteristics of Lactococcus lactis subsp. lactis CF4MRS in Artemia franciscana: a Biological Approach for the Control of Edwardsiellosis in Larviculture


Predominance of beneficial bacteria helps to establish a healthy microbiota in fish gastrointestinal system and thus to reduce emerging pathogen. In this study, the colonization efficacy of Lactococcus lactis subsp. lactis CF4MRS in Artemia franciscana and its potential as a probiotic in suppressing Edwardsiella sp. infection were investigated in vivo. The colonization extent of the bioencapsulated L. lactis was established through visualization of gfp gene-transformed L. lactis in A. franciscana. Here, we demonstrate that when A. franciscana is administrated with L. lactis at 108 CFU mL−1 for 8 h, the highest relative percentage of survival (RPS = 50.0) is observed after inoculation with Edwardsiella sp. The total counts of L. lactis entrapped in Artemia were the highest (ranged from 3.2 to 5.1 × 108 CFU mL−1), when 108–109 CFU mL−1 of L. lactis was used as starting inoculum, with the bioencapsulation performed within 8–24 h. Fluorescent microscopy showed gfp-transformed L. lactis colonized the external trunk surfaces, mid-gut and locomotion antennules of the A. franciscana nauplii. These illustrations elucidate the efficiency of colonization of L. lactis in the gastrointestinal tract and on the body surfaces of Artemia. In conclusion, L. lactis subsp. lactis CF4MRS shows a good efficacy of colonization in Artemia and has the potential for biocontrol/probiotic activity against Edwardsiella sp. infection.

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The authors would like to express their gratitude to Dr. Paloma López (Centro de Investigaciones Biológicas, CSIC, Madrid, Spain) for her technical guidance in gfp gene insertion and bacterial electrotransformation. Special thanks to Professor Sharr Azni Harmin (Universiti Selangor, Malaysia) and Associate Professor Yau Yan Lim (Monash University, Malaysia) for technical consultation.


This study was funded by the Higher Degree Research (HDR) grant from Monash University, Malaysian campus.

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Correspondence to Jiun Yan Loh.

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Loh, J.Y., Kay, G.L. & Ting, A.S.Y. Bioencapsulation and Colonization Characteristics of Lactococcus lactis subsp. lactis CF4MRS in Artemia franciscana: a Biological Approach for the Control of Edwardsiellosis in Larviculture. Mar Biotechnol 20, 353–362 (2018).

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  • Artemia franciscana
  • Biocontrol
  • Edwardsiella sp.
  • Green fluorescent protein
  • Lactococcus lactis subsp. lactis