Bioencapsulation and Colonization Characteristics of Lactococcus lactis
subsp. lactis CF4MRS in Artemia franciscana: a Biological Approach
for the Control of Edwardsiellosis in Larviculture
Jiun Yan Loh
Gemma L. Kay
Adeline Su Yien Ting
Received: 22 September 2016 /Accepted: 29 November 2017 /Published online: 13 April 2018
Springer Science+Business Media, LLC, part of Springer Nature 2018
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 10
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 × 10
), when 10
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.Inconclusion,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.
Keywords Artemia franciscana
Green fluorescent protein
Lactococcus lactis subsp. lactis
Aquaculture is the fastest growing food industry (Saravanan
et al. 2013). However, the rapid expansion of the industry has
resulted in massive occurrence of various fish diseases.
Aquaculture of fish, in particularly, turbot Scophthalmus
pia Oreochromis niloticus and catfish Clarias batrachus,of-
ten develop diseases such as edwardsiellosis (Sahoo et al.
1998; Kim et al. 2003; Zheng et al. 2004;Padrosetal.
2006). The Gram-negative pathogen Edwardsiella sp. is the
major cause of edwardsiellosis, which can cause hemorrhagic
septicemia including lesions on the skin, muscles and internal
organs in many aquatic species (Bullock and Herman 1985;
Mohanty and Sahoo 2007). The spread of Edwardsiella sp.
in fish culture systems can be transmitted via contaminated
equipment and facilities used in cultivation, or during the
harvesting process of the live feed (Mainous et al. 2010).
Common live feed include Artemia and rotifers and are
often used as natural food sources for post-larvae in finfish
or shrimp hatchery. Studies showed that Artemia could be a
potential bio-vehicle for the transmission of several fish
diseases; evidence indicates these live feeds can be in-
volved (directly or indirectly) in transmitting piscine cryp-
tosporidiosis infection caused by Cryptosporidium molnari
and Cryptosporidium scophthalmi (Méndez-Hermida et al.
Electronic supplementary material The online version of this article
(https://doi.org/10.1007/s10126-018-9813-9) contains supplementary
material, which is available to authorized users.
* Jiun Yan Loh
Faculty of Applied Sciences, UCSI University, No. 1, Jalan Menara
Gading, UCSI Heights, Cheras, 56000 Kuala Lumpur, Malaysia
School of Science, Monash University Malaysia, Jalan Lagoon
Selatan, 47500 Bandar Sunway, Selangor, Malaysia
Medical Microbiology Research Laboratory, Norwich Medical
School, University of East Anglia, Norwich NR4 7UQ, UK
Marine Biotechnology (2018) 20:353–362