183 
Identification of larval sea basses 
( Centropristis spp.) 
using ribosomal DNA-specific molecular assays* 
Abstract — The identification of sea 
bass ( Centropristis ) larvae to spe- 
cies is difficult because of similar 
morphological characters, spawning 
times, and overlapping species ranges. 
Black sea bass ( Centropristis striata) 
is an important fishery species and 
is currently considered to be over- 
fished south of Cape Hatteras, North 
Carolina. We describe methods for 
identifying three species of sea bass 
larvae using polymerase chain reac- 
tion (PCR) and restriction fragment 
length polymorphism (RFLP) assays 
based on species-specific amplifica- 
tion of rDNA internal transcribed 
spacer regions. The assays were 
tested against DNA of ten other co- 
occurring reef fish species to ensure 
the assay’s specificity. Centropristis 
larvae were collected on three cruises 
during cross-shelf transects and were 
used to validate the assays. Seventy- 
six Centropristis larvae were assayed 
and 69 (91%) were identified success- 
fully. DNA was not amplified from 
5% of the larvae and identification 
was inconclusive for 3% of the larvae. 
These assays can be used to identify 
sea bass eggs and larvae and will help 
to assess spawning locations, spawn- 
ing times, and larval dispersal. 
Manuscript submitted on 10 October 2007. 
Manuscript accepted on 6 February 2008. 
Fish. Bull. 106:183-193 (2008). 
The views and opinions expressed or 
implied in this article are those of the 
author and do not necessarily reflect 
the position of the National Marine 
Fisheries Service, NOAA. 
Mark W. Vandersea (contact author ) 1 
R. Wayne Litaker 1 
Katrin E. Marancik 2 
Jonathan A. Hare 2 
Harvey J. Walsh 3 
Email address for Mark W. Vandersea: 
Mark.W. Vandersea @ noaa.gov 
1 National Ocean Service, NOAA 
Center for Coastal Fisheries and 
Habitat Research 
101 Pivers Island Road 
Beaufort, North Carolina 28516 
2 National Marine Fisheries Service, NOAA 
28 Tarzwell Drive Narragansett Laboratory 
Narragansett, Rhode Island 02882 
3 Woods Hole Oceanographic Institution 
Marine Research Facility 217, MS#50 
Woods Hole, Massachusetts 02543 
A fundamental requirement of early 
life stage studies is the ability to iden- 
tify individuals to species. Consider- 
able knowledge of the eggs and larvae 
of most fish species in temperate and 
boreal ecosystems has been achieved 
(Kendall and Matarese, 1994; Berrien 
and Sibunka, 1999). In contrast, in 
subtropical and tropical ecosystems, 
the larvae of many species cannot 
be identified (Kendall and Matarese, 
1994), even for groups of important 
fishery species. For example, of the 73 
species in the snapper-grouper complex 
that range within U.S. federal waters, 
larvae of only one-half can be identi- 
fied to species and eggs of most spe- 
cies are undescribed (Richards, 2006). 
As anthropogenic stress on marine 
ecosystems continues to increase, new 
approaches to early life stage identi- 
fication are needed to obtain critical 
information about exploited species 
and to improve the scientific basis 
for conservation. For example, im- 
munological methods have been used 
successfully to identify invertebrate 
and fish larvae (Miller et ah, 1991; 
Paugam et ah, 2000; Garland and 
Zimmer, 2002; Taylor, 2004). Simi- 
Siya Lem 4 
Melissa A. West 5 
David M. Wyanski 6 
Elisabeth H. Laban 1 
Patricia A. Tester 1 
4 Duke University Medical Center 
PO. Box 2628 
Durham, North Carolina 27710 
5 David Clark Laboratory 
North Carolina State University 
PO Box 7617 
Raleigh, North Carolina 27695-7617 
6 Marine Resources Research Institute 
South Carolina Department of Natural Resources 
PO. Box 12559 
Charleston, South Carolina 29422-2559 
larly, mitochondrial DNA sequences 
(Hare et al., 1994; Pegg et ah, 2006) 
and restriction fragment length poly- 
morphisms (RFLP) have been used to 
identify fish eggs, larvae, and adults 
(Daniel and Graves, 1994; Aranishi et 
al., 2005; Hyde et al., 2005). Though 
highly accurate, these methods have 
yet to be used widely as part of stan- 
dard protocols for identifying fish 
larvae collected during research and 
monitoring surveys. 
We examined the application of 
molecular techniques for identifying 
species within the genus Centropristis 
(sea bass). Five species of Centropris- 
tis are currently recognized in the 
western North Atlantic Ocean. In ad- 
dition to black sea bass (C. striata), 
there are the following: bank sea bass 
(C. ocyurus), rock sea bass (C. phila- 
delphica), twospot sea bass (C. fuscu- 
la), and C. rufus. Black sea bass is an 
important commercial and recreation- 
al species on the northeast and south- 
east U.S. continental shelf. Along 
* Contribution number 635 of the South Car- 
olina Marine Resources Center, Charles- 
ton, South Carolina 29422-2559. 
