Genotyping Of Lactococcus garvieae Isolated From Rainbow Trout By RAPD-PCR Using M13 And P5 Primers In Turkey
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1
Uludag University, Department of Aquatic Animal Diseases, Türkiye
Aquaculture production is considerably large and total aquaculture production was reached 260.000 tons in 2015, in Turkey. Rainbow trout (Oncorhynchus mykiss) production is about %50 of these total fish production. Especially, when water temperature is above 15 ° C, Lactococcosis infection occurs in rainbow trout farms (L. garvieae, synonym Enterococcus seriolicida). The aim of this study was to evaluate the efficiency of randomly amplified polymorphic DNA (RAPD) method and to compare the discriminatory powers of M13 and P5 primers for genotyping L. garvieae isolates.
In this study, L. garvieae isolated on rainbow trout farms in different regions (Marmara, Aegean, Central Anatolia, Southeast and Eastern Anatolia) of Turkey in 2013-2015 years. Isolated bacteria were identified by classic microbiologic tests (Gram staining, oxidase, catalase, Oxidation/Fermentation), rapid identification kits (API 32 STREPT) and molecular analysis (PCR). RAPD PCR analysis was also applied for determining epidemiologic and genotypic relationship of 137 L. garvieae.
137 strains have been identified by PCR analyses. Strains were separated into groups by analyzing RAPD patterns. While 5 different genotypes have been identified by using M13 primer which determined 95 isolates (69.3%) were found similar profiles, 4 different genotypes have been identified using P5 primer which determined 96 isolates (70 %) were found similar profiles. Aegean and Central Anatolia regions’ strains found genetically similar to L. garvieae strains. While there were five different genetic groups of L. garvieae isolates in 2013, two different genetic groups were found isolates in 2014-2015 with M13 primer.
As a result; similarity of L. garvieae strains which isolated from trout farms in Turkey have found at least 69% with M13 and P5 primers. When RAPD-PCR analysis results were compared with M13 and P5 primers, it was concluded that M13 primer had more binding sites on DNA and discriminatory power of the M13 primers was found to be higher than P5.
Keywords:
Lactococcus garvieae,
genotyping,
RAPD-PCR,
M13 primer,
P5 primer
Conference:
AquaEpi I - 2016, Oslo, Norway, 20 Sep - 22 Sep, 2016.
Presentation Type:
Poster
Topic:
Aquatic Animal Epidemiology
Citation:
DUMAN
M,
ALTUN
S,
SATICIOĞLU
IB and
BÜYÜKEKİZ
AG
(2016). Genotyping Of Lactococcus garvieae Isolated From Rainbow Trout By RAPD-PCR Using M13 And P5 Primers In Turkey.
Front. Vet. Sci.
Conference Abstract:
AquaEpi I - 2016.
doi: 10.3389/conf.FVETS.2016.02.00017
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Received:
30 May 2016;
Published Online:
14 Sep 2016.
*
Correspondence:
MD, PhD. Izzet B SATICIOĞLU, Uludag University, Department of Aquatic Animal Diseases, Bursa, Türkiye, iburcinsat@uludag.edu.tr