International network for natural sciences – research journal
  • mendeley icon
  • linkedin icon
  • google plus icon
  • twitter icon
  • google scholar icon
  • facebook icon

Administration of live-attenuated vaccine of Vibrio harveyi to improve survival of gnotobiotic brine shrimp (Artemia salina) model against multipleVibrio infection

By: Y.K. Chin, Nurhidayu Al-saari, Aslizah Mohd Aris, Mohd Zamri-Saad, Ina- M.Y. Salwany

Key Words: Vibriosis, Artemia salina, Live-attenuated vaccine, Vibrio, Serine protease.

Int. J. Biosci. 14(4), 1-14, April 2019.


[Generate Certificate]


Newly developed live-attenuated protease derivative from pathogenic Vibrio harveyis train Vh1 as a live vaccine to against Vibriosis of aquatic animals. In the current study, we used the gnotobiotic A. salina as model to evaluate the safety and efficacy of the live-attenuated. This study was conducted by bacterial safety experiment and bacterial efficacy experiment. During the bacterial safety, the wild type and live-attenuated of V. harveyi (MVh-vhs) were tested for 48 hours on the Artemia larvae (instar II). During the efficacy experiment, the A. salina larvae were incubated with different concentration of live-attenuated V. harveyi (MVh-vhs), then challenged with V. harveyi, V. alginolyticus and V. parahaemolyticus. The result of safety experiment showed that the high concentration of live-attenuated mutant V. harveyi (MVh-vhs) at concentration of 109 CFU/mL is safe and had improved the A. salina larvae survival compared to other groups. On the other hand, pathogenic wildtype V. harveyi caused lethal effect on A. salina larvae by decreasing their survival. The surprising result of efficacy experiment showed that 107 CFU/mL of live attenuated MVh-vhs with 6 hours post incubation withA. salina larvae contributed higher survival while 109 CFU/mL of live attenuated MVh-vhs with 24 hours incubated A. salina larvae contributed higher survival against multiple Vibrio challenge. In this study, we concluded that the incubation time had affect bacterial concentration uptake by A. salina larvae and affect the effectiveness of Artemia bioencapsulation for targeted hosts.

| Views 259 |

| Views 259 |

Administration of live-attenuated vaccine of Vibrio harveyi to improve survival of gnotobiotic brine shrimp (Artemia salina) model against multipleVibrio infection

Aris AM. 2018. Development of Vibrio harveyi Protease Deletion Mutant as Live attenuated Vaccine against Vibriosis In Epinephelusfuscoguttatus (FORSSKAL, 1775) (Unpublished Doctoral Thesis). Universiti Putra Malaysia, Serdang, Selangor, Malaysia.

Aris AM, Ina-Salwany MY, Zamei-Saad M, Hassan MD, Norfarrah MA. 2016. Molecular characterization of Vibrio harveyi virulence- associated serine protease and outer membrane protein genes for vaccine development.  International Journal of Bioscience 8(3), 10-28, March 2016

Chatterjee S, Haldar S. 2012. Vibrio Related Disease in Aquaculture and Development of Rapid and Accurate Identification Methods. Journal of Marine Science: Research & Development 2012, S1, 002.

Destoumieux-Garzon D, Saulnier D, Garnier J, Jouffery C, Bulet P, Bachere E. 2001. Crustacean Immunity Antifungal Peptides are generated from C Terminus of Shrimp Hemcyanin in response to microbial challenge. The Journal of Biological Chemistry 276, 47070-47077.

Fazeli Z, Azari-Takami G. 2006. The Best time and Concentration for Yeast probiotic Enrichment of Artemia urmiana Nauplii. Pakistan Journal of Biological Sciences 9(11), 2159-2161. ISSN 1028-8880.

Gelabert R. 2002. Bioencapsulation in Artemia: II.  Influences of the particle concentration in the

enrichment process. Aquaculture 216 (2003), 143-153.

Gomez-Gil B, Herrera-Vega MA, Abreu-Grobois FA, Roque A. 1998. Bioencapsulation of Two Different Vibrio Species in Nauplii of the Brine Shrimp (Artemia franciscana). Applied and Environmental Microbiology 64(6), p 2318-2322 0099-2240/98/$04.00+0

Haenen OLM, Fouz B, Amaro C, Isern MM, Mikkelsen H, Zrncic S, Travers MA, Renault T, Wardle R, Hellstrom A, Dalsgaard I. 2014. Vibriosis in aquaculture.16th EAFP Conference, Tampere, Finland, 4th September 2013. Bulletin- European Association of Fish Pathologists 34(4), 138-148.

Hai NV, Buller N, Fotedar R. 2010. Encapsulation capacity of Artemia nauplii with customized probiotics for use in the cultivation of western king prawns (Penaeus latisuicatusKishinouye, 1896). Aquaculture Research, 41, 893-903.

Hauton C. 2012. The scope of the crustacean immune system for disease control. Journal of Invertebrate Pathology 110(2), June 2012, Pages 251-260.

Hu YH, Deng T, Sun BG, Sun L. 2012. Development and efficacy of an attenuated Vibrio harveyi vaccine candidate with cross protectivity against Vibrio alginoltyticus. Fish & Shellfish Immunology 32(2012), 1155-1161.

Huang YC, Jian JJ, Lu YS, Cai SH, Wang B, Tang JF, Pang HY, Ding Y, Wu ZH. 2012. Draft Genome Sequence of the Fish Pathogen Vibrio harveyi Strain ZJ0603. Journal of Bacteriology. December 2012. 194(23), 6644-6645.

Immanuel G. 2016. Bioencapsulation of Brine shrimp Artemia Nauplii with Probionts and Their Resistance Against Vibrio Pathogens. Journal of Fisheries and Aquatic science. ISSN 1816-4927.

Interaminense JA, Calazans NF, Caritas do Valle B, Vogeley JL, Peixoto S, Soares R, Lima Filho JV. 2014. Vibrio spp. Control at Brine Shrimp, Artemia, Hatching and Enrichment. Journal of the World Aquaculture Society.

Joosten PHM, Aviles-Trigueros M, Sorgeloos P, Rombout JHWM. 1995. Oral vaccination of juvenile carp (Cyprinus carpio) and gilthead seabream (Sparus aurata) with bioencapsulated Vibrio anguillarumbacterin. Fish and Shellfish Immunology (1995) 5, 289-299.

Kim SY, Chung HY, Lee DH, Lim JG, Kim, SK, Ku HJ, Kim YT, Kim H, Ryu S, Lee JH, Choi SH. 2016. Complete genome sequence of Vibrio parahaemolyticus strain FORC 008, a foodborne pathogen from a flounder fish in South Korea. Pathogens and Disease 74, 2016, FEMS.

Le Moullac G, Haffner P. 2000. Environmental factors affecting immune response in Crustacea. Aquaculture 191, 121-131.

Lee CY, Cheng MF, Yu MS, Pan MJ. 2002. Purification and characterization of a putative virulence factor, serine protease, from Vibrio parahaemolyticus. FEMS Microbiology Letters 209, 31-37.

Lee MN, Kim SK, Li XH, Lee JH. 2014. Bacterial virulence analysis using brine shrimp as an infection model in relation to the importance of quorum sensing and proteases. The Journal of General and Applied Microbiology 60, 169-174.

Lee SY, Soderhall K. 2002. Early events in crustacean innate immunity. Fish and Shellfish Immunology 12(5), May 2002, Pages 421-437.

Lin BC, Wang Z, Malanoski AP, O’Grady EA, Wimpee CF, Vuddhakul V, Alves N, Jr, Thompson FL, Gomez-Gil B, Vora GJ. 2010. Comparative genomic analyses identify the Vibrio harveyi genome sequenced strains BAA-1116 and HY01 as Vibrio campbellii. Wiley Environmental Microbiology Reports. 2010 Feb; 2(1), 81-89.

Liu L, Ge MF, Zheng XY, Tao Z, Zhou SM, Wang GL. 2016. Investigation of Vibrio alginolyticus, V. harveyi, and V. parahaemolyticus in large yellow croaker, Pseudosciaenacrocea (Richardson) reared in XiangshanBay, China. Aquaculture Report 3(2016), 220-224.

Liu YL, Zhao Y, Dai ZM, Chen HM, Yang WJ. 2009. Formation of Diapause Cyst Shell in Brine Shrimp, Artemia parthenogenetica, and its Resistance Role in Environmental Stresses. Journal of Biological Chemistry. 2009 jun 19, 284(25), 16931-16938.

Makridis P, Fjellheim AJ, Skjermo J, Vadstein O. 2000. Control of the bacterial flora of Brachiomusplicatilis and Artemia franciscana by incubation in bacterial suspensions. Aquaculture 185, 207-218.

Marques A, Thanh TH, Sorgeloos P, Bossier P, 2006. Use of microalgae and bacteria to enhance protection of gnotobiotic Artemia against different

pathogens. Aquaculture 258, 116-128.

Miyoshi SI. 2013. Extracellular proteolytic enzymes produced by human pathogenic species. Frontiers in Microbiology 4, 339.

Mutoloki S, Munang’andu HM, Evensen O. 2015. Oral vaccination of fish – antigen preparations, uptake, and immune induction. Frontier in Immunology 6, 519.

Nehlah R, Ina-Salwany MY, Zulperi Z. 2016. Antigenicity Analysis and Molecular Characterization of Two Outer Membrane Proteins of Vibrio alginolyticus Strain VA2 as Vaccine Candidates in Tiger Grouper Culture. Journal of Biological sciences 16.

Niu YF, Defoirdt T, Baruah K, Van de Wiele T, Dong SL, Bossier P. 2014. Bacillus sp. LT3 improves the survival of gnotobiotic brine shrimp (Artemia franciscana) larvae challenged with Vibrio campbellii by enhancing the innate immune response and by decreasing the activity of shrimp-associated Vibrios. Veterinary Microbiology 173, 279-283.

Patil PK, Gopel C, Panigrahi A, Rajababu D, Pillai SM. 2013. Oral administration of formalin killed Vibrio anguillarum cells improves growth and protection against challenge with Vibrio harveyi in banana shrimp. Letters in Applied Microbiology 58, 213-218.

Reis DB, Acosta NG, Almansa E, Navarro JC, Tocher DR, Andrade JP, Sykes AV, Rodriguez C. 2017. Comparative study on fatty acid metabolism of early stages of two crustacean species: Artemia sp. matanauplii and Grapsusadscensionis zoeae, as live prey for marine animals. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology. Volume 204, February 53-60.

Shahabi V, Maciag PC, Rivera S, Wallecha A. 2010. Live, attenuated strains of Listeria and Salmonella as vaccine vectors in cancer treatment Bioengineered Bugs. 2010 Jul-Aug; 1(4), 235-239.

Soltanian S, Thai QT, Dhont J, Sorgeloss P, Bossier P. 2007. The protective effect against Vibrio campbellii in Artemia nauplii by pure B-glucan and isogenic yeast cells differing in B-glucan and chitin content operated with a source-depended time lag. Fish and Shellfish immunology 23, 1003-1014.

Stappen GV. 1998. Morphology and life cycle. Biology and ecology of Artemia. Introduction, biology and ecology of Artemia. Manual on the Production and Use of Live Food for Aquaculture. FAO. Rome 1996. ISBN 92-5-1103934-8.

Steidler L. 2003. Genetically engineered probiotics. Best Practice & Research Clinical Gastroenterology. 17(5), 861-876.

Sung YY, Dhaena T, Defoirdt T, Boon N, MacRae TH, Sorgeloos P, Bossier P. 2009. Ingestion of bacteria overproducing DnaK attenuates Vibrio infection of Artemia franciscana larvae. Cell Stress and Chaperones 14, 603-609.

Tkavc R, Ausec L, Oren A, Gunde-Cimerman N. 2010. Bacteria associated with Artemia spp. along the salinity gradient of the solar salterns at Eilat (Israel). FEMS Microbiology Ecology 77(2).

Vadstein O, Bergh O, Gatesoupe F, Galindo-Villegas J, Mulero V, Picchietti S, Scapigliati G, Makridis P, Olsen Y, Dierckens K, Defoirdt, T, Boon N, De Schryver P, Bossier P. 2013. Microbiology and Immunology of fish larvae. Reviews in Aquaculture (2013) 5(1), S1-S25.

Vazzquez L, Alpuche J, Maldonado G, Agundis C, Pereyra-Morales A, Zenteno E. 2009. Review: Immunity mechanisms in crustaceans. Innate Immunity.

Verschuere L, Heang H, Criel G, Sorgeloos P, Verstraete W. 2000. Selected Bacterial Strains Protect Artemia spp. from the Pathogenic Effects of Vibrio proteolyticus CW8T2. Applied and Environmental Microbiology 66(3), 1139–1146.

Verschuere L, Rombaut G, Huys G, Dhont J, Sorgeloos P, Verstraete W. 1999. Microbial Control of the Culture of Artemia Juveniles through Preemptive Colonization by selected Bacterial Strains. Applied and Environmental Microbiology, June 1999. 65(6), p 2527-2533. PMCID: PMC91373

Interaminense JA, Calazans NF, do Valle BC, Vogeley JL, Peixoto S, Soares R, Lima JV. 2014. Vibrio spp. control at brine shrimp, Artemia, hatching and enrichment. Journal of the World Aquaculture Society 45(1), 65-74.

Y.K. Chin, Nurhidayu Al-saari, Aslizah Mohd Aris, Mohd Zamri-Saad, Ina- M.Y. Salwany.
Administration of live-attenuated vaccine of Vibrio harveyi to improve survival of gnotobiotic brine shrimp (Artemia salina) model against multipleVibrio infection.
Int. J. Biosci. 14(4), 1-14, April 2019.
Copyright © 2019
By Authors and International Network for
Natural Sciences (INNSPUB)
innspub logo
english language editing
    Publish Your Article
    Submit Your Article
Email Update