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.
KeywordsArtemia franciscana Biocontrol Edwardsiella sp. Green fluorescent protein Lactococcus lactis subsp. lactis
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.
- Amend DF (1981) Potency testing of fish vaccines. Dev Biol Stand 49:447–454Google Scholar
- Andani HRR, Tukmechi A, Meshkini S, Sheikhzadeh N (2012) Antagonistic activity of two potential probiotic bacteria from fish intestines and investigation of their effects on growth performance and immune response in rainbow trout (Oncorhynchus mykiss). J Appl Ichthyol 2012:1–7Google Scholar
- APHA (1985) Standard methods for the examination of water and wastewater, 16th edn. American Public Health Association, Washington, D.C.Google Scholar
- Badhul Haq MA, Vijayasanthi P, Vignesh R, Shalini R, Somnath Chakraborty R, Rajaram R (2012) Effect of probiotics against marine pathogenic bacteria on Artemia franciscana. J Pharmacol Sci 2(4):38–43Google Scholar
- Bullock GL, Herman RL (1985) Edwardsiella infections of fishes. US Fish and Wildlife Service, University of Nebraska-Lincoln, pp 6Google Scholar
- Cunningham F, Lees P, Elliott J (2010) In: F. Cunningham et al (Eds) Comparative and veterinary pharmacology. Handbook of experimental pharmacology. Springer-Verlag Berlin, HeidelbergGoogle Scholar
- FAO (Food and Agriculture Organization of the United Nations) & WHO (World Health Organization) (2001) Evaluation of health and nutritional properties of powder milk and live lactic acid bacteria. Food and Agriculture Organization of the United Nations and World Health Organization expert consultation report, FAO, Rome, ItalyGoogle Scholar
- Hai NV, Buller N, Fotedar R (2010) Encapsulation capacity of Artemia nauplii with customized probiotics for use in cultivation of western king prawns (Penaeus latisulcatus Kishinouye, 1968). Aquac Res 41:893–903Google Scholar
- Kim KW, Wang X, Choi SM, Park GJ, Koo JW, Bai SC (2003) No synergistic effects by the dietary supplementation of ascorbic acid, a-tocopheryl acetate and selenium on the growth performance and challenge test of Edwardsiella tarda in fingerling Nile tilapia, Oreochromis niloticus L. Aquac Res 34:1053–1058Google Scholar
- Mohanty BR, Sahoo PK (2007) Edwardsiellosis in fish: a brief review. J Biosci 32(7):1331–1344Google Scholar
- Padros F, Zarza C, Dopazo L, Cuadrado M, Crespo S (2006) Pathology of Edwardsiella tarda infection in turbot, Scophthalmus maximus (L.). J Fish Dis 29:87–94Google Scholar
- Sahoo PK, Mukherjee SC, Sahoo SK (1998) Aeromonas hydrophila versus Edwardsiella tarda: A pathoanatomical study in Clarias batrachus. Aquaculture 6:57–66Google Scholar
- Saravanan K, Nilavan SE, Sudhagar SA, Naveenchandru V (2013) Diseases of mariculture finfish species: a review. Isr J Aquacult Bamidgeh. IJA_64.2012.831Google Scholar
- Zheng D, Mai K, Liu S, Cao L, Liufu Z, Xu W, Tan B, Zhang W (2004) Effect of temperature and salinity on virulence of Edwardsiella tarda to Japanese flounder, Paralichthys olivaceus (Temminck et Schlegel). Aquac Res 35:494–500Google Scholar