Abstract
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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References
Amend DF (1981) Potency testing of fish vaccines. Dev Biol Stand 49:447–454
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 2012:1–7
APHA (1985) Standard methods for the examination of water and wastewater, 16th edn. American Public Health Association, Washington, D.C.
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):38–43
Bullock GL, Herman RL (1985) Edwardsiella infections of fishes. US Fish and Wildlife Service, University of Nebraska-Lincoln, pp 6
Chang CI, Liu WY (2002) An evaluation of two probiotic bacterial strains, Enterococcus faecium SF 68 and Bacillus toyoi, for reducing edwardsiellosis in cultured European eel, Anguilla anguilla L. J Fish Dis 25:311–315
Colorni A (1985) A study on the bacterial flora of giant prawn, Macrobrachium rosenbergii, larvae fed with Artemia salina nauplii. Aquaculture 49:1–10
Cunningham F, Lees P, Elliott J (2010) In: F. Cunningham et al (Eds) Comparative and veterinary pharmacology. Handbook of experimental pharmacology. Springer-Verlag Berlin, Heidelberg
Dornan S, Collins MA (1990) High efficiency electroporation of Lactococcus lactis subsp. lactis LM0230. Letter Appl Microbiol 11:62–64
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, Italy
Fernández de Palencia P, Nieto C, Acebo P, Espinosa M, López P (2000) Expression of green fluorescent protein in Lactococcus lactis. FEMS Microbiol Lett 183:229–234
Fjellheim AJ, Klinkenberg G, Skjermo J, Aasen IM, Vadstein O (2010) Selection of candidate probionts by two different screening strategies from Atlantic cod (Gadus morhua L.) larvae. Vet Microbiol 144:153–159
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 41:893–903
Harikrishnan R, Balasundaram C, Heo MS (2010) Lactobacillus sakei BK19 enriched diet enhances the immunity status and disease resistance to streptococcosis infection in kelp grouper, Epinephelus bruneus. Fish Shellfish Immun 29:1037–1043
Holo H, Nes IF (1989) High-frequency transformation, by electroporation, of Lactococcus lactis subsp. cremoris grown with glycine in osmotically stabilized media. Appl Environ Microbiol 55(12):3119–3123
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 34:1053–1058
Lamari F, Sadok K, Bakhrouf A, Gatesoupe FJ (2013) Selection of lactic acid bacteria as candidate probiotics and in vivo test on Artemia nauplii. Aquacult Int 22:699–708
Lauzon HL, Gudmundsdottir S, Pedersen MH, Budde BB, Gudmundsdottir BK (2008) Isolation of putative probionts from cod rearing environment. Vet Microbiol 132:328–339
Loh JY, Lim YY, Harmin SA, Ting ASY (2014) In vitro assessment on intestinal microflora from commonly farmed fishes for control of the fish pathogen Edwardsiella tarda. Turk J Vet Anim Sci 38:257–263
Loh JY, Ting ASY (2016) Effects of potential probiotic Lactococcus lactis subsp. lactis on digestive enzymatic activities of live feed Artemia franciscana. Aquacult Int 24:1341–1351
Mainous ME, Smith SA, Kuhn DD (2010) Effect of common aquaculture chemicals against Edwardsiella ictaluri and E. tarda. J Aquat Anim Health 22:224–228
Marques A, François JM, Dhont J, Bossier P, Sorgeloos P (2004) Influence of yeast quality on performance of gnotobiotically grown Artemia. J Exp Mar Biol Ecol 310:247–264
Méndez-Hermida F, Gómez-Couso H, Ares-Mazás E (2007) Possible involvement of Artemia as live diet in the transmission of cryptosporidiosis in cultured fish. Parasitol Res 101(3):823–827
Mohanty BR, Sahoo PK (2007) Edwardsiellosis in fish: a brief review. J Biosci 32(7):1331–1344
Motlagh HRZ, Farhangi M, Rafiee G, Noori F (2012) Modulating gut microbiota and digestive enzyme activities of Artemia urmiana by administration of different levels of Bacillus subtilis and Bacillus licheniformis. Aquacult Inter 20:693–705
Padros F, Zarza C, Dopazo L, Cuadrado M, Crespo S (2006) Pathology of Edwardsiella tarda infection in turbot, Scophthalmus maximus (L.). J Fish Dis 29:87–94
Phatarpekar PV, Kenkre VD, Sreepada RA, Desai UM, Achuthankutty CT (2002) Bacterial flora associated with larval rearing of the giant freshwater prawn, Macrobrachium rosenbergii. Aquaculture 203:279–291
Ruiz-Cruz S, Solano-Collado V, Espinosa M, Bravo A (2010) Novel plasmid-based genetic tools for the study of promoters and terminators in Streptococcus pneumoniae and Enterococcus faecalis. J Microbiol Methods 83:156 ̶ 163
Ruiz-Masό JA, Lόpez-Anguilar C, Nieto C, Sanz M, Burόn P, Espinosa M, del Solar G (2012) Construction of a plasmid vector based on the pMV158 replicon for cloning and inducible gene expression in Streptococcus pneumonia. Plasmid 67:53–59
Sahul Hameed AS (1993) A study of the aerobic heterotrophic bacterial flora of hatchery-reared eggs, larvae and post-larvae of Penaeus indicus. Aquaculture 117:195–204
Sahoo PK, Mukherjee SC, Sahoo SK (1998) Aeromonas hydrophila versus Edwardsiella tarda: A pathoanatomical study in Clarias batrachus. Aquaculture 6:57–66
Saravanan K, Nilavan SE, Sudhagar SA, Naveenchandru V (2013) Diseases of mariculture finfish species: a review. Isr J Aquacult Bamidgeh. IJA_64.2012.831
Situmorang ML, Dierckens K, Mlingi FT, Van Delsen B, Bossier P (2014) Development of a bacterial challenge test for gnotobiotic Nile tilapia Oreochromis niloticus larvae. Dis Aquat Org 109:23–34
Sivakumar VK, Sarathi M, Venkatesan C, Sivaraj A, Sahul Hameed AS (2009) Experimental exposure of Artemia to Hepatopancreatic parvo-like virus and subsequent transmission to post-larvae of Penaeus monodon. J Invertebr Pathol 102:191–195
Touraki M, Karamanlidou G, Koziotis M, Christidis I (2013) Antibacterial effect of Lactococcus lactis subsp. lactis on Artemia fransciscana and Dicentrarchus labrax larvae against the fish pathogen Vibrio anguillarum. Aquacult Inter 21:481–495
Villamil L, Figueras A, Planas M, Novoa B (2010) Pediococcus acidilactici in the culture of turbot (Psetta maxima) larvae: administration pathways. Aquaculture 307:83–88
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 35:494–500
Ziaei-Nejad S, Rezaei MH, Takami GA, Lovett DL, Mirvaghefi AR, Shakouri M (2006) The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activitiy, survival and growth in the Indian white shrimp (Fenneropenaeus indicus). Aquaculture 252:516–524
Acknowledgements
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.
Funding
This study was funded by the Higher Degree Research (HDR) grant from Monash University, Malaysian campus.
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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). https://doi.org/10.1007/s10126-018-9813-9
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DOI: https://doi.org/10.1007/s10126-018-9813-9