Determination of virulence associated immunogenic proteins in some of Lactococcus garvieae strains

Document Type : Original Article


Department of Fisheries Technology Engineering, Surmene Faculty of Marine Sciences, Karadeniz Technical University, Trabzon, Turkey


Lactococcosis disease incident caused by Lactococcus garvieae has been increased with increasing aquaculture productions and outbreaks of the disease have become a threat on farmed species. To prevent lactococcosis, inactivated vaccine has been used, however, it only provides protection when given by injection. Other than inactivated vaccine, various vaccines such as subunit vaccines can be developed. In the present study, total protein profile of 43 strains of L. garvieae isolated from fish, milk and cheese by SDS-PAGE and virulence associated immunogenic proteins of L. garvieae strains using western blot with hyper-immune rabbit sera were determined. After analyzing whole-cell lysate protein of L. garvieae strains with SDS-PAGE, protein bands were ranged between 8.00 and 140.00 kDA. Among strains, variable protein bands were ranged between 17.00 and 48.00 kDa with some variability in the staining intensity of the protein bands and formed in 6 clusters. The immunogenic protein bands were ranged between 25.00 - 75.00 kDa. Only a variable and highly immunogenic protein band was observed between 40.00 and 45.00 kDa. Most of the strain including Lgper had 44.00 kDa immunogenic protein while nonvirulent ATCC strain had 42.50 kDA immunogenic protein. Predominant immuno-reactive proteins encoded by genes can be used as a subunit vaccine.


Main Subjects


    1. Ozturk RC, Altinok I. Bacterial and viral fish diseases in Turkey. Turk J Fish Aquat Sci 2014; 14: 275-297.
    2. Austin B, Austin DA. Bacterial fish pathogens; diseases of farmed and wild fish. 5th ed. London, UK: Springer2012; 17-58.
    3. Plumb JA, Hanson LA. Health maintenance of cultured fishes: Principal microbial diseases. 3rd ed. Iowa, USA: Wiley-Blackwell 2011; 483.
    4. Ture M, Altinok I, Capkin E. Comparison of pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus PCR and biochemical tests to characterize Lactococcus garvieae. J Fish Dis 2015; 38(1): 37-47.
    5. Park KH, Matsuoka S, Nakai T, et al. A virulent bacteriophage of Lactococcusgarvieae (formerly Enterococcusseriolicida) isolated from yellowtail Seriola quinqueradiata. Dis Aquat Organ 1997; 29: 145-149.
    6. Chang PH, Lin CW, Lee YC. Lactococcus garvieae infection of cultured rainbow trout, Oncorhynchus mykiss, in Taiwan and associated biophysical characteristics and histopathology. B Eur Assoc Fish Pat 2002; 22(5): 319-327.
    7. Kang SH, Shin GW, Shin YS, et al. Experimental evaluation of pathogenicity of Lactococcus garvieae in black rockfish (Sebastes schlegeli). J Vet Sci 2004; 5(4): 387-390.
    8. Ghasemi SM, Bouzari M, Yoon BH, et al. Comparative genomic analysis of Lactococcus garvieae phage WP-2, a new member of Picovirinae subfamily of Podoviridae. Gene 2014; 551(12): 222-229.
    9. Diler O, Altun S, Adiloglu AK, et al. First occurrence of Streptococcosis affecting farmed rainbow trout (Oncorhynchus mykiss) in Turkey. B Eur Assoc Fish Pat 2002; 22(1): 21-26.
    10. Eldar A, Goria M, Ghittino C, et al. Biodiversity of Lactococcus garvieae strains isolated from fish in Europe, Asia, and Australia. Appl Environ Microbiol 1999; 65(3): 1005-1008.
    11. Ravelo C, Magarinos B, Herrero MC, et al. Use of adjuvanted vaccines to lengthen the protection against lactococcosis in rainbow trout (Oncorhynchus mykiss). Aquaculture 2006; 251(2-4): 153-158.
    12. Altinok I. Multiplex PCR assay for detection of four major bacterial pathogens causing rainbow trout disease. Dis Aquat Organ 2011; 93(3): 199-206.
    13. Soltani M, Mohamadian S, Ebrahimzahe-Mousavi HA, et al. Shirazi thyme (Zataria multiflora) essential oil suppresses the expression of the epsD capsule gene in Lactococcus garvieae, the cause of lactococcosis in farmed fish. Aquaculture 2014; 433: 143-147.
    14. Kawanishi M, Yoshida T, Yagashiro S, et al. Differences between Lactococcus garvieae isolated from the genus Seriola in Japan and those isolated from other animals (trout, terrestrial animals from Europe) with regard to pathogenicity, phage susceptibility and genetic characterization. J Appl Microbiol 2006; 101(2): 496-504.
    15. Ooyama T, Hirokawa Y, Minami T, et al. Cell-surface properties of Lactococcus garvieae strains and their immunogenicity in the yellowtail Seriola quinqueradiata. Dis Aquat Organ 2002; 51(3): 169-177.
    16. Ture M, Altinok I. Detection of putative virulence genes of Lactococcus garvieae. Dis Aquat Organ 2016; 119(1): 59-66.
    17. Sanchez L, Martinez P, Vinas A, et al. Analysis of the structure and variability of nucleolar organizer regions of Salmo-Trutta by C-, Ag-, and restriction endonuclease banding. Cytogenet Cell Genet 1990; 54(1-2): 6-9.
    18. Wickstrom D, Wagner S, Baars L, et al. Consequences of depletion of the signal recognition particle in Escherichia coli. J Biol Chem 2011; 286(6): 4598-4609.
    19. Ding Y, Fujimoto LM, Yao Y, et al. Solid-state NMR of the Yersinia pestis outer membrane protein Ail in lipid bilayer nanodiscs sedimented by ultracentrifugation. J Biomol NMR 2015; 61(3-4): 275-286.
    20. Kobayashi A, Kobayashi Y, Shiomi K. Fish allergy in patients with parvalbumin-specific immunoglobulin E depends on parvalbumin content rather than molecular differences in the protein among fish species. Biosci Biotechnol Biochem 2016; 80(10): 2018-2021.
    21. Altun S, Adiloglu A, Kubilay A, et al. Immunogenic and antigenic profiles of nine Lactococcus garvieae strains from different rainbow trout farms. Isr J Aquacult-Bamid 2007; 59(2): 111-116.
    22. Huang HY, Chen YC, Wang PC, et al. Efficacy of a formalin-inactivated vaccine against Streptococcus iniae infection in the farmed grouper Epinephelus coioides by intraperitoneal immunization. Vaccine 2014; 32(51): 7014-7020.
    23. Winkelstroter LK, Tulini FL, De Martinis ECP. Identification of the bacteriocin produced by cheese isolate Lactobacillus paraplantarum FT259 and its potential influence on Listeria monocytogenes biofilm formation. Lwt-Food Sci Technol 2015; 64(2): 586-592.
    24. Hunter PR, Gaston MA. Numerical Index of the discriminatory ability of typing systems - an application of simpsons index of diversity. J Clin Microbiol 1988; 26(11): 2465-2466.
    25. Descheemaeker P, Pot B, Ledeboer AM, et al. Comparison of the Lactococcus lactis differential medium (Dcl) and SDS-Page of whole-cell proteins for the identification of Lactococci to subspecies level. Syst Appl Microbiol 1994; 17(3): 459-466.
    26. Dos Santos O, De Resende MCC, De Mello AL, et al. The use of whole-cell protein profile analysis by SDS-PAGE as an accurate tool to identify species and subspecies of coagulase-negative staphylococci. APMIS 2012; 120(1): 39-46.
    27. Raissy M, Ansari M. Antibiotic susceptibility of Lactococcus garvieae isolated from rainbow trout (Oncorhynchus mykiss) in Iran fish farms. Afr J Biotechnol 2011; 10(8): 1473-1476.
    28. Ture M, Boran H. Phenotypic and genotypic antimicrobial resistance of Lactococcus sp, strains isolated from rainbow trout (Oncorhynchus mykiss). Bull Vet Inst Pulawy 2015; 59: 37-42.
    29. Madureira P, Baptista M, Vieira M, et al. Streptococcus agalactiae GAPDH is a virulence-associated immunomodulatory protein. J Immunol 2007; 178(3): 1379-1387.
    30. Tsai MA, Wang PC, Cao TT, et al. Immunoprotection of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Lactococcus garvieae against Lactococcosis in tilapia. J Gen Appl Microbiol 2013; 59(6): 437-449.
Volume 10, Issue 2
June 2019
Pages 101-107
  • Receive Date: 10 July 2018
  • Revise Date: 05 September 2018
  • Accept Date: 25 September 2018
  • First Publish Date: 15 June 2019