Isolation and Characterization of Lactococcus garvieae from Diseased Rainbow Trout (Oncorhynchus mykiss, Walbaum) Cultured in Northern Iran Based on the Nucleotide Sequences of the 16s rRNA Gene

Milad ADEL; Atefe Esmailian DEHKORDI; Zahra YAGHOUBZADEH; Elham Khalili SADRABAD; Alireza BABAALIAN

Authors

Milad ADEL Department of Aquatic Animal Health and Diseases, Caspian Sea Ecology Research Center, Sari
Atefe Esmailian DEHKORDI DVM Graduate, Faculty of Veterinary Medicine, University of Shahrekord, Shahrekord
Zahra YAGHOUBZADEH Department of Aquatic Animal Health and Diseases, Caspian Sea Ecology Research Center, Sari
Elham Khalili SADRABAD Department of Health and Food Quality Control, Faculty of Veterinary Medicine, University of Shahrekord, Shahrekord
Alireza BABAALIAN Department of Aquatic Animal Health and Diseases, Veterinary Organization, Sari

Keywords:

Lactococcus garvieae, Oncorhynchus mykiss, 16s rRNA, PCR, Iran

Abstract

This study was done to determine the molecular and biochemical identification of some causative agents of lactococcosis in farmed rainbow trout in Mazandaran provenience (northern Iran). A total of 200 moribund rainbow trout, suspected of lactococcosis from 10 rainbow trout farms in Mazandaran province, were collected during spring 2012 to winter 2012. Sampling was done from the kidney, spleen, liver and brain and cultured aseptically onto brain heart infusion (BHI) agar plates and incubated at 25 °C for 24 - 48 h. Results of bacteriological cultures of these organs showed 19 % Lactococcus garvieae (38 fish), 9 % Streptococcus spp., (18 fish), 17 % Yersinia spp. (36 fish), and 55 % of fish were culture negative. The PCR assay was developed based on the 16s rRNA gene of L. garvieae for the rapid and specific detection and identification of this pathogen from different sources. Two pairs of primers were designed based on the nucleotide sequences of the 16s rRNA gene of L. garvieae. After PCR assay on isolated bacterial colonies, DNAs extracted from 38 L. garvieae gave the expected 1107 bp PCR fragment of 16S rDNA sequences, which is specific for L. garvieae. The results of this study suggest the use of molecular methods along with current biochemical methods are effective diagnostic tools in the identification of L. garvieae. The combination of these methods for diagnosis of other bacterial disease is recommended.

doi:10.14456/WJST.2015.76

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

Author Biography

Milad ADEL, Department of Aquatic Animal Health and Diseases, Caspian Sea Ecology Research Center, Sari

Department of Aquatic Animal Health and Diseases

References

B Austin and DA Austin. Bacterial fish pathogens, diseases of farmed and wild fish. Vol I. Chichester, UK, Springer Praxis Publishing, 2007, p. 123-9.

R Russo, H Mitchell and RPE Yanong. Characterization of Streptococcus iniae isolated from ornamental cyprinid fishes and development of challenge models. Aquaculture 2006; 256, 105-10.

C Shoemaker, JJ Evans and PH Klesius. Density and dose: factors affecting mortality of Streptococcus iniae infected tilapia, Oreochromis niloticus. Aquaculture 2000; 188, 229-35.

M Akhlaghi and M Keshavarzi. The occurrence of streptococcosis in the cultured rainbow trout of Fars province. Iran. J. Vet. Res. 2002; 2, 183-9.

M Soltani, Sh Jamshidi and I Sharifpour. Streptococcosis caused by Streptococcus iniae in farmed rainbow trout (Onchorhynchus mykiss) in Iran: biophysical characteristics and pathogensis. Bull. Eur. Assoc. Fish. Pathol. 2005; 25, 95-107.

F Fadaeifard, H Momtaz, E Rahimi and A Mirzakhani. Detection of Streptococcus iniae and Lactococcus garvieae by multiplex polymerase chain reaction (PCR) in some rainbow trout farms of Iran. Afr. J. Biotechnol. 2012; 11, 260-3.

E Rahimi-Kia and Y Mehrabi. Detection and identification of different Streptococcosis strains in farmed rainbow trout in Boyerahmad and Dena regions (North South of Iran). World J. Fish & Marine Sci. 2013; 5, 315-21.

H Sharifiyazdi, M Akhlaghi, M Tabatabaei and SM Mostafavizadeh. Isolation and characterization of Lactococcus garvieae from diseased rainbow trout (Oncorhynchus mykiss, Walbaum) cultured in Iran. Iran. J. Vet. Res. 2010; 11, 342-50.

M Soltani, GH Nikbatht, H Mousavi and N Ahmadzadeh. Epizootic outbreak of lactococcosis caused by Lactococcus garvieae in farmed rainbow trout (Oncorhynchus mykiss) in Iran. Bull. Eur. Assoc. Fish. Pathol. 2008; 28, 207-12.

MG Carvalho, MC Vianni, JA Elliot, M Reeves, RR Facklam and LM Teixeira. Molecular analysis of Lactococcus garvieae and Enterococcus gallinarum isolated from water buffalos with subclinical mastitis. Adv. Exp. Med. Biol. 1997; 418, 401-10.

LA Devriese, J Hommez, H Laevens, P Baneadme and F Haesebrouck. Identification of aesculin hydrolyzing streptococci and enterococci from subclinical intramammary infections in dairy cows. J. Vet. Microbiol. 1999; 70, 87-94.

K Rantsiou, R Urso, L Iacumin, C Cantoni, P Cattaneo and G Comi. Culture-dependent and independent methods to investigate the microbial ecology of Italian fermented sausages. Appl. Environ. Microbiol. 2005; 71, 1977-86.

JJ Fefer, KR Ratzan, SE Sharp and E Saiz. Lactococcus garvieae endocarditic: report of a case and review of the literature. Microb. Infect. Dis. 1998; 32, 127-30.

M Soltani, M Hazeri, I Sharifpour, S Mirzargar and P Shohre. Study of Bacterial Diseases in Farmed Rainbow Trout (Oncorhynchus mikyss) in Mazandaran Province. Iran. J. Vet. Microbiol. 2012; 8, 1-12.

M Soltani and M Tarahomi. Study of streptococcosis/lactococcosis in some farmed rainbow trout in Fars province. In: Proceedings of the 1th International Congress on Aquatic Animal Health Management & Diseases, Tehran, Iran, 2009. p. 231-2.

M Soltani, M Hazeri, I Sharifpour, S Mirzargar and P Shohre. Study of bacterial diseases in farmed rainbow trout (Oncorhynchus mikyss) in Mazandaran province. Iran. J. Vet. Microbiol. 2012; 8, 1-12.