DAO 76:151-161 (2007)  -  doi:10.3354/dao076151

Use of the 16S-23S rDNA internal transcribed spacer of Roseovarius crassostreae for epizootiological studies of juvenile oyster disease (JOD)

Aaron P. Maloy1,3, Bruce J. Barber2,4, Katherine J. Boettcher1,5,*

1Department of Biochemistry, Microbiology and Molecular Biology, and 2School of Marine Sciences, University of Maine, Orono, Maine 04469, USA
3Present address: Centre of Applied Marine Biotechnology, Letterkenny Institute of Technology, Letterkenny, County Donegal, Ireland
4Present address: Terra Environmental Services, Inc., 5215 W. Laurel Street, Suite 110, Tampa, Florida 33607-1728, USA
5Present address: 11208 Beechwood Pointe, Smithfield, Virginia 23430, USA
*Corresponding author. Email:

ABSTRACT: Juvenile oyster disease (JOD) in Crassostrea virginica is caused by the marine bacterium Roseovarius crassostreae. Although the 16S rRNA genes of the bacterial isolates exhibit little variation, 2 genetic signatures (GSI and GSII) may be discerned by Ava I digestion of the 16S-23S internal transcribed spacer (ITS). In this study we analyzed isolates from JOD epizootics throughout the northeastern USA (including affected adults for the first time) to better understand how oyster populations encounter and become affected by the pathogen. Isolates from a given epizootic usually had the same ITS signature; however, the involvement of both genetic signatures was occasionally detected, even within the same oyster. Sequencing was used to localize the variable Ava I site to a 100 bp region of low sequence identity, and detection of additional base changes resulted in the identification of 11 distinct genotypes. One genotype was found only in Martha’s Vineyard, Massachusetts, USA and persisted in JOD survivors. Two genotypes were associated with Maine epizootics, and both were believed to be unique to that region until 2004, when one was detected in Martha’s Vineyard among oysters that had survived colonization by the local genotype. Apparent competition between those 2 genotypes was also detected among a population of juveniles. Five genotypes were found only in New York, and the other 3 were isolated from both New York and from around Cape Cod, Massachusetts. Relationships between the geographic occurrence and phylogenetic relatedness of genotypes were compared with regional current patterns to identify possible mechanisms controlling their distribution.

KEY WORDS: Juvenile oyster disease · JOD · Roseovarius oyster disease · ROD · Crassostrea virginica · Roseovarius crassostreae · Internal transcribed spacer · ITS · Epizootiology

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