Genetic characterization of Lactococcus garvieae isolated from farmed rainbow trout by random amplified polymorphic DNA-PCR in Iran


F. FADAEIFARD
M. RABIEI
M. F. SHARIFPOUR
Abstract

Lactococcosis is one of the main bacterial infections of fish around the world. Lactococcus garvieae has been a major cause of rainbow trout losses in freshwater farming. This study aimed to genotype and determine the variability of L. garvieae isolated from infected farmed rainbow trout in Iran by the RAPD-PCR method. Bacterial samples were collected from 12 farms located in the western part of Iran and suspect ted to carry Lactococcus infection. Two hundred bacterial cultures containing cocci shaped bacteria were cultured in Trypticase soy agar (TSA) and blood agar mediums. All bacterial cultures were tested by conventional microbiological and biochemical tests, and PCR assay to identify L. garvieae by 16S rDNA genes. The RAPD-PCR method was used to determine the genetic patternof all isolates. The sample strain pattern of the isolates was analyzed in the NTSYS program. According to a similarity coefficient index of 70%, all L. garvieae isolates were separated into two groups with four RAPD profile types. The highest and the lowest genetic pairwise similarity among the isolates were 98% and 54%, respectively. The results of the present study revealed that RAPD-PCR is an applicable method to describe the genetic diversity of different strains of L .garvieae among farmed fish.

Article Details
  • Sezione
  • Research Articles
Downloads
I dati di download non sono ancora disponibili.
Riferimenti bibliografici
Altun S, Buyukekiz A, Onuk EE, Duman M, and Ciftci A (2013) Phenotypic, genotypic characterisation and antimicrobial susceptibility determination of Lactococcus garvieae strains. Kafkas Univ Vet Fak 19 (3): 375-381.
Altun S, Diler O, Adiloglu AK (2004) Genotyping of Lactococcus garvieae strains from rainbow trout (Oncorhynchus mykiss) by 16S Rdna sequencing. Bull. Eur. Ass. Fish Pathol 24: 119-124.
Austin B, Austin DA (2012) Bacterial Fish Pathogens. Disease of Farmed and Wild Fish, Fifth Edition, Springer 88:99-106.
Belkum VA, Meis J (1994) Polymerase chain reaction mediated DNA fingerprinting in bacterial epidemiology. Clin Infect Dis 18: 1017-1018.
Brunt J, Austin B (2005): Use of a probiotic to control lactococcosis and streptococcosis in rainbow trout, Oncorhynchus mykiss (Walbaum). J Fish Dis 28: 693-701.
Chan JF1, Woo PC, Teng JL, Lau SK, Leung SS, Tam FC, Yuen KY (2011) Primary infective spondylodiscitis caused by Lactococcus garvieae and a review of human L. garvieae infections. Infection 39: 259−264.
Elliot JA, Collins MD, Pigott NE, Facklam RR (1991) Differentiation of Lactococcus lactis and Lactococcus garvieae from humans by comparison of whole-cell protein patterns. J Clin Microbiol 20: 2731-4.
Fadaeifard F, Momtaz H, Rahimi E, Mirzakhani A (2012) Detection of Streptococcus iniae and Lactococcus garvieae by multiplex polymerase chain reaction (PCR) in some rainbow trout farms of Iran. Afr J Biotechnol 11(2):260-263.
Ferrario C, Ricci G, Milani C, Lugli GA, Ventura M, Eraclio G, Eraclio G, Borgo F, Fortina MG (2013) Lactococcus garvieae: Where Is It From? A First Approach to Explore the Evolutionary History of This Emerging Pathogen. PLoS ONE 8(12): e84796.
Findik A, İça T, Onuk EE, Percin D, Kevenk TO, Ciftci A (2011) Molecular typing and cdt gene prevalence of Campylobacter jejuni isolates from various sources. Trop Anim Health Prod 43: 711-719.
Fortina MG, Ricci G, Foschino R, Picozzi C, Dolci P, Zeppa G, Cocolin L, Manachini PL (2007) Phenotypic typing, technological properties and safety aspects of Lactococcus garvieae strains from dairy environments. J Appl Microbiol 103: 445-453.
Foschino R, Nucera D, Volponi G, Picozzi C, Ortoffi M, Bottero MT (2008) Comparison of Lactococcus garvieae strains isolated in northern Italy from dairy products and fishes through molecular typing. J Applied Microbiol 105: 652-662.
Gibello A, Galán-Sánchez F, Blanco MM, Rodríguez-Iglesias M, Domínguez L, Fernández-Garayzábal JF (2016) The zoonotic potential of Lactococcus garvieae: an overview on microbiology, epidemiology, virulence factors and relationship with its presence in foods. Res Vet Sci 109:59−70.
Kusuda K, Kawai K, Salati F, Banner CR, Fryer JL (1991) Enterococcus seriolicida sp. nov., a fish pathogen. Int J Syst Bacteriol 41:406-409.
Li WK, Chen YS, Wann SR, Liu YC, Tsai HC (2008) Lactococcus garvieae endocarditis with initial presentation of acute cerebral infarction in a healthy immunocompetent man. Intern Med 47:1143-1146.
Magarinos B, Toranzo AE, Barja JL, Romalde JL (2000) Existence of two geographically-linked clonal lineages in the bacterial fish pathogen Photobacterium damselae subsp. piscicida evidenced by random amplified polymorphic DNA analysis. Epidemiol. Infect 125: 213-219.
Meyburgh CM, Bragg RR, Boucher CE (2017) Lactococcus garvieae: An emerging bacterial pathogen of fish. Dis Aquat Organ 123: 67-79.
Ravelo C, Magariños B, Lόpez-Romalde S, Toranzo AE, Romalde JL (2003) Molecular fingerprinting of fish-pathogenic Lactococcus garvieae strains by random amplified polymorphic DNA analysis. J Clin Microbiol 41: 751-6.
Sharifiyazdi H, Akhlaghi M, Tabatabaei M, Mostafavizadeh SM (2010) Isolation and characterization of Lactococcus garvieae from diseased rainbow trout (Oncorhynchus mykiss, Walbaum) cultured in Iran. Iran J Vet Res. 11: 342−350.
Soltani M, Jamshidi SH, Sharifpour I (2005) Streptococcosis caused by Streptococcus iniae in farmed rainbow trout (Oncorhynchus mykiss) in Iran:Biophysical characteristics and pathogenesis. Bull Eur AssAltun S, Buyukekiz A, Onuk EE, Duman M, and Ciftci A (2013) Phenotypic, genotypic characterisation and antimicrobial susceptibility determination of Lactococcus garvieae strains. Kafkas Univ Vet Fak 19 (3): 375-381.
Altun S, Diler O, Adiloglu AK (2004) Genotyping of Lactococcus garvieae strains from rainbow trout (Oncorhynchus mykiss) by 16S rDNA sequencing. Bull. Eur. Ass. Fish Pathol 24: 119-124.
Austin B, Austin DA (2012) Bacterial Fish Pathogens. Disease of Farmed and Wild Fish, Fifth Edition, Springer 88:99-106.
Belkum VA, Meis J (1994) Polymerase chain reaction mediated DNA fingerprinting in bacterial epidemiology. Clin Infect Dis 18: 1017-1018.
Brunt J, Austin B (2005): Use of a probiotic to control lactococcosis and streptococcosis in rainbow trout, Oncorhynchus mykiss (Walbaum). J Fish Dis 28: 693-701.
Chan JF1, Woo PC, Teng JL, Lau SK, Leung SS, Tam FC, Yuen KY (2011) Primary infective spondylodiscitis caused by Lactococcus garvieae and a review of human L. garvieae infections. Infection 39: 259−264.
Elliot JA, Collins MD, Pigott NE, Facklam RR (1991) Differentiation of Lactococcus lactis and Lactococcus garvieae from humans by comparison of whole-cell protein patterns. J Clin Microbiol 20: 2731-4.
Fadaeifard F, Momtaz H, Rahimi E, Mirzakhani A (2012) Detection of Streptococcus iniae and Lactococcus garvieae by multiplex polymerase chain reaction (PCR) in some rainbow trout farms of Iran. Afr J Biotechnol 11(2):260-263.
Ferrario C, Ricci G, Milani C, Lugli GA, Ventura M, Eraclio G, Eraclio G, Borgo F, Fortina MG (2013) Lactococcus garvieae: Where Is It From? A First Approach to Explore the Evolutionary History of This Emerging Pathogen. PLoS ONE 8(12): e84796.
Findik A, İça T, Onuk EE, Percin D, Kevenk TO, Ciftci A (2011) Molecular typing and cdt gene prevalence of Campylobacter jejuni isolates from various sources. Trop Anim Health Prod 43: 711-719.
Fortina MG, Ricci G, Foschino R, Picozzi C, Dolci P, Zeppa G, Cocolin L, Manachini PL (2007) Phenotypic typing, technological properties and safety aspects of Lactococcus garvieae strains from dairy environments. J Appl Microbiol 103: 445-453.
Foschino R, Nucera D, Volponi G, Picozzi C, Ortoffi M, Bottero MT (2008) Comparison of Lactococcus garvieae strains isolated in northern Italy from dairy products and fishes through molecular typing. J Applied Microbiol 105: 652-662.
Gibello A, Galán-Sánchez F, Blanco MM, Rodríguez-Iglesias M, Domínguez L, Fernández-Garayzábal JF (2016) The zoonotic potential of Lactococcus garvieae: an overview on microbiology, epidemiology, virulence factors and relationship with its presence in foods. Res Vet Sci 109:59−70.
Kusuda K, Kawai K, Salati F, Banner CR, Fryer JL (1991) Enterococcus seriolicida sp. nov., a fish pathogen. Int J Syst Bacteriol 41:406-409.
Li WK, Chen YS, Wann SR, Liu YC, Tsai HC (2008) Lactococcus garvieae endocarditis with initial presentation of acute cerebral infarction in a healthy immunocompetent man. Intern Med 47:1143-1146.
Magarinos B, Toranzo AE, Barja JL, Romalde JL (2000) Existence of two geographically-linked clonal lineages in the bacterial fish pathogen Photobacterium damselae subsp. piscicida evidenced by random amplified polymorphic DNA analysis. Epidemiol. Infect 125: 213-219.
Meyburgh CM, Bragg RR, Boucher CE (2017) Lactococcus garvieae: An emerging bacterial pathogen of fish. Dis Aquat Organ 123: 67-79.
Ravelo C, Magariños B, Lόpez-Romalde S, Toranzo AE, Romalde JL (2003) Molecular fingerprinting of fish-pathogenic Lactococcus garvieae strains by random amplified polymorphic DNA analysis. J Clin Microbiol 41: 751-6.
Sharifiyazdi H, Akhlaghi M, Tabatabaei M, Mostafavizadeh SM (2010) Isolation and characterization of Lactococcus garvieae from diseased rainbow trout (Oncorhynchus mykiss, Walbaum) cultured in Iran. Iran J Vet Res. 11: 342−350.
Soltani M, Jamshidi SH, Sharifpour I (2005) Streptococcosis caused by Streptococcus iniae in farmed rainbow trout (Oncorhynchus mykiss) in Iran:Biophysical characteristics and pathogenesis. Bull Eur Ass Fish Pathol. 25: 95-106.
Soltani M, Nikbatht GH, Mousavi HAE, Ahmadzadeh N (2008) Epizootic outbreak of lactococcosis in farmed rainbow trout (Oncorhynchus mykiss) in Iran. Bull Eur Ass Fish Pathol 8: 207-212.
Tsai MA, Wang PC, Liaw LL, Yoshida T, Chen SC (2012) Comparison of genetic characteristics and pathogenicity of Lactococcus garvieae isolated from aquatic animals in Taiwan. Dis Aquat Org 102: 43−51.
Vela AI, Vázquez J, Gibello A, Blanco MM, Moreno MA, Liébana P, Albendea C, Alcalá B, Mendez A, Domínguez L, Fernández-Garayzábal JF (2000) Phenotypic and genetic characterization of Lactococcus garvieae isolated in Spain from lactococcosis outbreaks and comparison with isolates of other countries and sources. J Clin Microbiol 38:3791-3795.
Vendrell D, Balcazar JL, Ruiz-Zarzuela I, de Blas I, Gironés O, Múzquiz JL (2006) Lactococcus garvieae in fish: a review. Comp Immun Microbiol Inf Dis 29: 177-198.
Villani F, Aponte M, Blaiotta G, Mauriello G, Pepe O, Moschetti G (2001) Detection and characterization of a bacteriocin, garviecin L1-5, produced by Lactococcus garvieae isolated from raw cow’s milk. J Appl Microbiol 90: 430-9.
Wang CYC, Shie HS, Chen SC, Huang JP, Hsieh IC, Wen MS, Lin FC, Wu D. (2007) Lactococcus garvieae infections in humans: possible association with aquaculture outbreaks. Int J Clin Pract 61: 68−73.
Welsh J, Mcclelland M(1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18: 7213-7218.
Woo PTK, Bruno DW (2011) Fish Diseases and Disorders III: Viral, Bacterial and Fungal Infections, 2nd edition, CABI publishing. Zlotkin A, Eldar A, Ghittino C, Ghittino C, Bercovier H (1998) Identification of Lactococcus garvieae by PCR. J Clin Microbiol 36: 983-5.