# Mapping and Characterization of Deep Sea Coral Ecosystems off the Coast of Florida 

**[Total Funds Requested:]{.smallcaps}** \$101,571 ( FY2005)
[**Category:** Deep Corals]{.smallcaps}

**PI:** Sandra Brooke, Oregon Institute of Marine Biology, PO Box 5389,
Charleston, OR 97420. Phone: 541-888-2581 x318, Fax: 541-888-3250,
Email: sbrooke@darkwing.uoregon.edu

**Co-PI:** John Reed, Harbor Branch Oceanographic Institution, 5600 U.S.
1 North, Fort Pierce, FL 34946. Phone: 772-465-2400 x205, Fax:
772-461-2221, Email: <jreed@hboi.edu>

**Co-PI:** Charles Messing, Nova Southeastern University, 8000 North
Ocean Drive, Dania Beach, Florida 33004. Phone: 954-262-3658, Fax:
954-262-4098, Email: <messingc@nsu.nova.edu>

With the continuing global depletion of coastal fisheries, commercial
fishing vessels are moving into deeper water and exploiting fish and
crustacean species that are frequently associated with complex coral
structures. Deep-sea coral ecosystems (DSCE) exist at numerous locations
around the coast of Florida. These coral communities provide habitat for
relatively unknown but highly diverse assemblages of fish and
invertebrates, many of which are undoubtedly undescribed species. There
is therefore an urgent need to identify these coral habitats, map their
distribution and abundance, and document any potentially valuable
fishery species associated with them. Resource managers can use this
information to develop conservation policy and scientists can use it as
a foundation for hypothesis driven ecological and physiological
research. Since deepwater reefs are not visible to the general public,
dissemination of information through the media and the education system
is vital in order to create empathy for their protection. The
overarching objective of this proposal is to increase the current
understanding of deep-water coral habitats along Florida's coasts.
Primary objectives use shipboard bathymetry and an ROV to map coral
distribution and characterize communities (particularly commercial
fisheries' species) found in association with high-relief geological
features in four regions of interest: the base of the Florida-Hatteras
Slope, the Miami and Pourtalès Terraces, and the SW Florida shelf slope.
Secondary objectives address aspects of coral biology, diversity and
biogeography of associated fauna, and microbial ecology. The proposed
research will not only provide important information about the deepwater
coral ecosystems off the Florida coast, but also contains a significant
outreach element.

**[Identification of the problem]{.smallcaps}**

Over the last several years, deep-sea corals have been moving to the
forefront of scientific and public awareness. Unfortunately, much of the
interest in deepwater reefs has been stimulated by the realization that
many of these fragile and largely unknown resources are currently being
impacted by destructive fishing practices faster than science can
allocate funds to describe them. With the continuing depletion of
coastal fisheries, commercial fishing vessels are moving into deeper
water and exploiting fish and crustacean species that frequently
associate with complex coral structures (Koslow and Gowlett-Jones 1998).
There is a global need to identify potential coral habitats, map their
distribution and abundance, and document any potentially valuable
fishery species associated with them. Resource managers can use this
information to develop conservation policy, and scientists can use it as
a foundation for hypothesis-driven ecological and physiological
research. Unlike shallow water tropical reefs, which are popular
vacation venues for millions of tourists annually, deep-water reefs are
not visible to the public. Dissemination of information through the
media and the education system is therefore vital in order to create
empathy for their protection. The research outlined in this proposal
will provide a strong foundation of information on the deepwater coral
ecosystems off the Florida coast and contains a significant
education/outreach element.

Deep-water topographic features are abundant at various locations off
the southeastern United States, including the base of the
Florida-Hatteras Slope, the Blake Plateau, the Straits of Florida, the
SW Florida slope and the eastern Gulf of Mexico. Deep-sea corals are
major components of the benthic assemblages associated with these
features, either as framework-builders of the features themselves, or as
primary generators of complex habitat for dense and diverse assemblages
of invertebrates and fishes. Deep-sea coral structures and their
associated fauna are often referred to as bioherms or lithoherms
(Teichert 1958, Stetson et al. 1962, Neumann et al. 1977, Wilson 1979,
Reed 1980, Friewald et al. 1997, Paull et al. 2000). Bioherms are
defined as ancient organic reefs of mound-like form built by a variety
of marine invertebrates. Some consist of caps of living coral covering
unconsolidated mud and coral debris. Lithoherms however, are high
relief, lithified carbonate limestone (Neumann et al. 1977), which
provide hard substrate for a variety of benthic fauna, including corals.
These topographic features occur extensively throughout the world's
oceans and have great potential resource value. However, their benthic
invertebrate and fish populations have only been described for a small
percentage of these deep-sea coral ecosystems (DSCE).

Extensive work has been conducted on the relatively shallow ahermatypic
*Oculina varicosa* reefs (75-100 m) off the central east coast (Reed
1981, 1983, 1992, Brooke 2002, Brooke and Young 2003), but information
on other DSCEs in Florida waters is limited to a few trawl and dredge
records (e.g. Cairns 1979), and fewer submersible observations (Neumann
& Ball 1970). Recent research expeditions by Reed et al. from Harbor
Branch Oceanographic Institution (HBOI), using submersibles and ROVs,
together with previous research in the 1990s and 1980s, have provided
new information on the status, distribution, habitat, and biodiversity
of some of these poorly understood deep reef ecosystems. Although
certain deep coral habitats, such as the *Oculina* banks, have been
documented as principal spawning sites for a variety of fishery species
(Gilmore and Jones 1992), life histories of fish species associated with
the deeper *Lophelia* systems have not yet been examined. It is likely
that Lophelia may provide spawning habitat for deeper reef species such
as lower slope roughies, (*Geophyroberyx darwinii*), wreckfish
(*Polyprion americanus*), groupers (*Epinephelus mystacinus, E.
nigritus, E. flavolimbatus and E. niveatus*), and snappers (*Etelis
oculatus, Lutjanus campechanus, L. buccanella, L. vivanus* and
*Pristopomoides aquilonaris*).

Biomedical expeditions in 2002 and 2003 funded by the National Oceanic
and Atmospheric Administration's Office of Ocean Exploration (OE)
initially explored additional deep-water reef sites in the western
Atlantic and eastern Gulf of Mexico. These were the first submersible
and ROV dives ever to document the habitat and benthic biodiversity of
some of these deep-water reefs. The dominant sessile macrobenthos here
were azooxanthellate, colonial scleractinian corals *Lophelia pertusa*,
*Madrepora oculata*, and *Enallopsammia profunda*; various hydrocorals
of the family Stylasteridae, Bamboo Corals of the family Isididae,
Antipatharian Black Corals and a wide variety of sponges. These provide
complex habitat for biologically rich and diverse communities of
associated fish, crustaceans, mollusks, echinoderms, polychaete and
sipunculan worms, anemones and other sessile and motile macrofauna, many
of which are undoubtedly undescribed species. Preliminary studies
discovered new species of octocorals and sponges from some of these
sites (Reed et al. 2004, submitted). Considerable work remains to
analyze and prepare these data for scientific publications.

In 2004, during a State of Florida funded mission with the
Johnson-Sea-Link (JSL) submersible, Reed et al. discovered nearly 300
potential targets during echosounder transects that may be deep-water
reefs off the east coast of Florida at depths of 732 m (2400 ft). Some
of these are up to 168 m (550 ft) in height (Reed 2004 in prep, Reed and
Wright 2004). However, these are preliminary analyses based on only a
few submersible or ROV dives at the various sites. The resource
potential for fisheries exploitation and presence of novel compounds
that may produce pharmaceutical drugs is unknown. Although these areas
are not currently designated as Marine Protected Areas (MPAs) or Habitat
Areas of Particular Concern (HAPCs), they are incredibly diverse and
irreplaceable resources. Activities involving bottom trawling,
pipelines, or oil/gas production could negatively impact these
environments

**[Justification]{.smallcaps}**

NOAA and the South Atlantic Fishery Management Council have requested
information on the state of knowledge of DSCE in the South Atlantic
region. Information on the distribution of DSCEs is needed to designate
new areas for protected status, because the continuing depletion of
coastal fisheries may cause fishermen to move to deeper habitats in
search of valuable commercial species such as royal red shrimp
(*Hymenopenaeus robustus*), Wreckfish (*Polyprion americanus*) or deep
water snappers and groupers. One of these species, the Warsaw Grouper,
*Epinephelus nigritus*, is a candidate for designation as a threatened
and endangered species. NOAA is currently developing sites for priority
mapping, and designation of deep-sea coral Marine Protected Areas. Data
compiled during the proposed project will provide information on
potential areas for designation of MPAs and HAPC's as well as areas of
scientific interest for future research.

An increased understanding of the ecology and biology of these
deep-water corals will provide insight into the influences that create
the different communities found in each region of the Florida shelf. The
great variation in *Lophelia* colony morphology or the dominance of
hydrocorals vs. scleractinians may be driven by food availability,
current speed or other environmental and/or biological factors. The
population genetics of *Lophelia* will provide information on larval
dispersal patterns, which reflect hydrography and may help interpret
distributions of other dispersive species. Finally, these deep-water
systems are at a disadvantage in gaining public empathy because most
people will never see them. It is the responsibility of scientists to
educate the public about these valuable resources. This proposal
therefore also includes an element of education/outreach toward
teachers, students and the public.

# Statement of objectives

As its overarching objective, this proposal will increase current
understanding of DSCEs along Florida's coasts. Previous explorations
have documented that these habitats are widespread in the region, but we
still know very little about the extent or distribution of the corals,
their ecology or their role as essential fish habitat, which are all
substantial justification for further effort. Primary objectives explore
the distribution and characterization of communities found in
association with lithoherms and other high relief, live-bottom,
geological features in the regions of interest. Secondary objectives
address aspects of coral biology, diversity of associated fauna, and
microbial ecology. Education and public outreach efforts are also a
significant aspect of this proposal.

.

**Primary objectives:**

1.  Map selected deep-water, high relief ecosystems that support coral
    communities. These sites include lithoherms and high-relief
    escarpments on the southwest Florida shelf and the east coast of
    Florida from Jacksonville to Miami. Specific sites will be selected
    from the four proposed regions described below.

2.  Characterize selected sites within the four regions using *in situ*
    observations, ROV transects, digital images and collections of
    macrofauna. Collect and inventory dominant cnidarians and sponges.

3.  Identify dominant fish species associated with coral communities,
    especially those appropriate for current or future fisheries
    exploitation. Document aggregations or spawning behavior.

4.  Collect and inventory other associated fauna from coral communities,
    and identify key species common to all regions and endemic to each.

5.  Describe the geology (e.g., substrate type, topography) and
    hydrography (e.g., prevailing current, water temperature) at each
    site, and identify the characteristic features of each region.

**Secondary objectives:**

1.  Collect samples of *L. pertusa* from each study site for analysis of
    reproductive status, growth rates, and colony morphology.

2.  Collect samples of *L. pertusa* for inclusion in a larger scale
    population genetic analysis of this species, currently being
    conducted through USGS.

3.  Characterize the microbial communities associated with colonies of
    *Lophelia* *pertusa* and other habitat-forming coral species,
    ambient water and surrounding sediment.

4.  Conduct preliminary investigations of the chemical richness of
    deepwater octocorals by analyzing selected species for
    concentrations and types of marine natural products.

5.  Provide opportunities for area teachers to accompany the research
    cruise, to learn about these deep-water environments, to develop
    classroom materials and a video record that will be incorporated
    into classroom learning experiences throughout the state. Develop a
    video record of the expedition for public consumption.

##  [Proposed study sites]{.smallcaps}

The sites for the proposed study fall within four regions of the Florida
shelf as described below. We have GPS coordinates and topographic
descriptions of eleven specific sites within Region 1, seven sites in
Region 2, seven sites in Region 3 and four sites in Region 4. Site
details are not presented in this proposal because some of the
information is proprietary and the tabulated information occupies
several pages. Additional sites will be chosen on station based on
cruise logistics and site bathymetry. The results of bathymetry surveys
will determine precise locations for ROV deployment. During the ROV
dives, both sonar and visual observations will be employed to
ground-truth geological features.  In regions 1-3 (east Florida coast,
Miami and Pourtalès Terraces and Straits), \~53 species of deep-water
Scleractinia (Cairns and Chapman 2002), about \~15 species of
stylasterids (Cairns 1986) and an unknown number of octocorals have been
recorded.  In Region 4, there are 84 species of deep-water Scleractinia
(Cairns and Chapman 2002, Cairns in prep), 5 stylasterids, and 115
octocorals (Cairns in prep). Although the number of octocorals is high,
this is the least known of these three cnidarian groups, and many new
taxa are to be expected.

**Region 1: East Florida Coast *Lophelia* Pinnacles**

Along a 222-km stretch along north and central eastern Florida (from
Jacksonville to Jupiter), nearly 300 mounds from 8 to 168 m in height
(25-550 ft) have been partially mapped by using a single beam
echosounder (Fig. 1). Between 1982 and 2004, the presence of *Lophelia*
mounds and lithoherms was confirmed using JSL and ROV dives (Reed 2002,
Reed and Wright 2004). The northern sites off Jacksonville appear to
have more lithoherm structure (Paull et al. 2000), with exposed rock,
whereas the features from south of St. Augustine to Jupiter were
comprised of *Lophelia* coral pinnacles or mud mounds capped with dense
1-m-tall thickets of *Lophelia* *pertusa* and *Enallopsammia profunda*
with varying amounts of coral debris and live coral. Dominant
habitat-forming coral species were *Lophelia pertusa, Madrepora
oculata*, *Enallopsammia profunda*, bamboo coral (Isididae), black coral
(Antipatharia), and diverse populations of other octocorals and sponges
(Reed 2004 in prep).

Figure 1. Height of *Lophelia* pinnacles and lithoherms on transects
from Jacksonville to Jupiter, Florida at depths of 600 to 800 m (from
Reed 2004 in prep).

> ![](media/image1.wmf)

**Region 2: Miami Terrace Escarpment**

The Miami Terrace is a 65-km long carbonate platform that lies between
Boca Raton and South Miami at depths of 200-400 m in the northeastern
Straits of Florida. It consists of high-relief Tertiary limestone
ridges, scarps and slabs that provide extensive hard bottom habitat
(Uchupi 1966, 1969, Kofoed and Malloy 1965, Uchupi and Emery 1967,
Malloy and Hurley 1970, Ballard and Uchupi 1971, Neumann and Ball 1970).
At the eastern edge of the Terrace, a high-relief, phosphoritic
limestone escarpment of Miocene age with relief of up to 90 m at depths
of 365 m is capped with *Lophelia* *pertusa*, stylasterids, bamboo
coral, and various sponges and other octocorals (Reed 2004 in prep, Reed
and Wright 2004). Dense aggregations of 50-100 wreckfish were observed
here during 11 JSL submersible dives in May 2004 (Reed in prep).
Previous studies in this region include geological studies on the Miami
Terrace (Neumann and Ball 1970, Ballard and Uchupi 1971) and dredge- and
trawl-based faunal surveys in the 1970s primarily by the University of
Miami (e.g. Halpern 1970, Holthuis 1971, 1974, Cairns 1979). *Lophelia*
mounds are also present at the base of the escarpment (\~670 m) within
the axis of the Straits of Florida, but we know little of their
distribution abundance or associated fauna (Neumann and Ball 1970).

**Region 3: Pourtalès Terrace Lithoherms**

The Pourtalès Terrace provides extensive, high-relief, hard-bottom
habitat, covering 3,429 km^2^ (1,000 nm^2^) at depths of 200-450 m. The
Terrace parallels the Florida Keys for 213 km and has a maximum width of
32 km (Jordan and Stewart 1961, Jordan et al. 1964, Land and Paull
2000). Reed et al. (2004 submitted) surveyed several deep-water,
high-relief, hard-bottom sites including the Jordan and Marathon
deep-water sinkholes on the outer edge of the Terrace, and five
high-relief bioherms on its central eastern portion. The JSL and Clelia
submersibles were used to characterize coral habitat and describe the
fish and associated macrobenthic communities. These submersible dives
were the first to enter and explore any of these features. The upper
sinkhole rims range from 175 to 461 m in depth and have a maximum relief
of 180 m. The Jordan Sinkhole may be one of the deepest and largest
sinkholes known. The high-relief bioherms (the proposed study sites
within this region) lie in 198 to 319 m, with a maximum height of 120 m.
A total of 26 fish taxa were identified from the sinkhole and bioherm
sites. Species of potential commercial importance include tilefish,
sharks, speckled hind, yellow-edge grouper, warsaw grouper, snowy
grouper, blackbelly rosefish, red porgy, drum, scorpion fish, amberjack,
and phycid hakes. Many different species of Cnidaria were recorded,
including Antipatharia, stylasterid hydrocorals, octocorals, and a
scleractinian (*Solenosmilia variabilis*). The benthic communities of
the Pourtalès Terrace bioherms differ from those of the lithoherms along
the northeastern Straits of Florida (Messing et al. 1990) chiefly in
that the Pourtalès Terrace communities lack reef-forming scleractinian
corals, (such as *Lophelia)*, and stalked crinoids (Reed et al. 2004
submitted). Dense thickets of stylasterid corals form the dominant
communities on top of these lithoherms. In addition, trawl collections
suggest that the Terrace supports a unique benthic assemblage of
invertebrates rarely found elsewhere in the Straits of Florida, e.g.,
the hydroid *Cladocarpus sigma*, and the unstalked crinoids *Comatonia
cristata* and *Coccometra hageni* (Bogle 1975, Meyer et al. 1978).

**Region 4: Southwest Florida Shelf *Lophelia* Lithoherms**

This region consists of dozens and possibly hundreds of 5-15 m tall
lithoherms at depths of 500 m, which may be capped with thickets of
*Lophelia* coral and rubble. In 1987, Newton et al. described the area
from a limited dredge and seismic survey. In 2003, Seabeam topographic
mapping was conducted over a small portion of the region (Fig. 2) (Reed
2004, Reed 2004 in prep), but ROV dives were made to ground-truth only
three of the features: a 36-m tall escarpment and two of the lithoherms.
The lithoherms appear to consist of rugged black phosphorite-coated
limestone boulders and outcrops capped with 0.5-1.0 m tall thickets of
*Lophelia pertusa*, which were approximately 10-20% alive. Dominant
sessile macrofauna include stony corals, octocorals, stylasterid
hydrocorals, black corals and sponges. The high number of hard-bottom
lithoherms revealed by the (limited) Seabeam mapping effort indicate
tremendous potential for unexplored coral and fish habitat in this
region. Interestingly, in bottom samples taken on the outer edge of the
southwest Florida shelf (159 m), Messing (unpublished) found an
unstalked crinoid assemblage not found elsewhere in Florida but
characteristic of deeper Cuban and Bahamian waters (\>300 m).

Fig. 2: Seabeam image of a southward view of the SW Florida Slope
lithoherms (x2 vertical exaggeration). Image courtesy of NOAA's Ocean
Exploration website, September 2003.

![](media/image2.png)

**[Proposed methodology]{.smallcaps}**

**Characterization of coral habitat**

Selection of the ROV launch site/bottom target site will be based on
both previously reported coral locations and the ship's bathymetric
survey. During the ROV dive, visual examination of the surrounding
environment will be used in conjunction with the ROV sonar and the
ship's bathymetry data to determine the distribution and extent of the
corals. In areas of well-developed coral cover, the different reef
structure categories will be identified (for example, isolated colonies,
live thickets, senescent reef, surrounding benthos), and standardized
video transects will be taken for community characterization. Ten to
twenty minutes of stationary video will be taken for several (\>5)
examples of each category to document associated fauna, preferably with
a low light camera with a minimal external light source. Close-up
digital images and (wherever possible) live samples will also be taken
for identification purposes. At low-density coral or non-coral sites,
appropriate habitat descriptor categories will be created (e.g.
sponge-dominated pavement) and similar observations of
habitat-associated fauna will be made. At each video station,
hydrological conditions such as temperature, current speed and
direction, salinity, and turbidity will be noted. Video footage will be
analyzed for habitat-specific biodiversity and species' distributions,
noting especially those species with resource value. Any sign or
indication of habitat damage or anthropogenic impact will also be noted.
The community association data will be compared with information from
other *Lophelia* systems, through collaborators in the USA and Europe as
part of an investigation into latitudinal differences of deep-water
coral reef community ecology.

**Identification of dominant coral-associated fish species**

Very little is known about the species, distribution or abundance of
fish associated with DSCE in Florida waters. This is a critical
information gap since the exploitation of commercially viable fish
populations poses the highest potential threat to these DSCE. Fish
populations will be classified from extensive video footage made during
the coral transects and collections made when possible depending on the
ROV technology available. Fish abundance and species distributions will
be spatially analyzed relative coral habitat distribution so that the
statistical significance of habitat association can be examined. This
will begin a first order documentation of fish association with each
type of DSCE. In addition, any aggregation or spawning behaviors will be
documented as fully as possible. This work will provide the basis for
further research efforts in defining fish communities and fishery
species associated with deep coral reefs around the lower Florida
peninsula. Prime fishery species at the chosen study sites are likely to
be roughies (trachichthyidae), wreckfish (polyprionidae), groupers
(serranidae) and snappers (lutjanidae), which have been previously
observed at depths of 300 to 700 m in the Florida Straits (Gilmore and
Jones pers. observations from JSL submersibles 1975-1997).

**Collection of coral and associated fauna**

Small discrete samples of live and dead coral colonies and
coral-associated fauna will be collected using the ROV hydraulic
manipulators. The samples will be maintained in separate containers and
at ambient temperature until they reach the deck for processing.
However, if this is not possible, the samples will be removed as quickly
as possible to avoid degradation. Corals and associated fauna will be
photographed *in situ* (at long range and close-up) then sampled and
cataloged according to museum standards. Additional coral samples will
be processed for each objective according to protocols described in the
methods. Observations of behavior and habitat utilization of
coral-associated fauna will be recorded in order to augment
morphological and molecular-based taxonomic descriptions. As many
species as possible will be identified on site; others will be
distributed to recognized taxonomic experts for identification and
ultimate deposit in major museum collections. Sponge systematics and
distributional ecology will be documented by Dr. Klaus Ruetzler of the
National Museum of Natural History. Specimens will be deposited in the
National Collections under his curation. Dr Jerry Harasewych, also of
the NMNH will identify mollusks, and deposit voucher specimens in the
National Collections. If a particular molluskan genus or species is
common and can be readily collected, Dr Harasewych will investigate
its population genetic structure, in order to compare the biogeography
of deep and shallow water mollusks from around the Florida peninsula and
to determine the depth range of the east Florida ecotone (Wise et al.
2004). If samples warrant, a continuing study of the geographic
variation and population genetics of pleurotomariids (Slit shells) will
be expanded, and novel taxa will be incorporated into several current
broad scale projects on gastropod phylogeny.

**Geological characterization**

A four-tiered scheme will be used to characterize the surficial geology
at and adjacent to each of the study sites, as follows: 1)
regional-setting at a scale of 5 to 10 km; 2) local-geomorphology at a
scale of 1 to 2 km; 3) study-site landscape at a scale of \<1 km; and 4)
study-site micro-habitats (e.g. survey transects, physical collections).
Regional-setting attributes will consist of the general shape and
orientation of the along‑isobath trend and the nature of the dominant
geomorphology of the upper continental slope province. Study-site
landscape descriptions will provide detailed depictions of the
bathymetry, geomorphology, and hard substrate. Geological features will
be identified and classified using direct visual observations and
substrate samples. Sediment samples will be collected from the study
sites using 30-cm steel push cores operated by the ROV's manipulator arm
and stored in a holder mounted on the front of the ROV. On deck, cores
will be sub-divided for analysis. A small part of each core will be
allocated for microbial DNA extractions and the remainder will be dried,
sieved into size fractions and weighed. Any infauna will be removed and
identified.

### 

### Determination of Scleractinian reproductive status 

Samples of *Lophelia* and other stony corals will be fixed for 24 h in a
solution of 5% formalin, washed in distilled water and decalcified in
10% hydrochloric acid for 10-12 hours. Polyps will then be dehydrated
through a series of ethanol concentrations, embedded in paraffin wax,
cut into 8µm sections, and stained using Mayer's Haematoxylin/Eosin B
staining techniques. Images of oocytes will be taken using an Optronics
digital camera attached to an Olympus BX50 compound microscope. Image
analysis software (UTHSCSA) will be used to measure and record the area
of each oocyte. The 'feret' diameter will be calculated and used in
statistical analysis. This estimates the diameter of a hypothetical
circle with the same area as the object measured. These values will
ultimately be compared with other samples collected during different
months to describe the gametogenic cycle of *Lophelia pertusa.* Several
decalcified polyps from each colony will be transferred to 30% ethanol
for fecundity analysis. Oocytes will be removed using fine forceps and
needles under a dissecting microscope. The polyp length and total number
of eggs per polyp will be recorded. This data will be compared with
information available for the North East Atlantic (Waller and Tyler in
press), Northern Gulf of Mexico (Brooke in prep.) and the Trodhejm
Fjords in Norway (Brooke and Jarnegren in prep.).

**Comparison of growth bands and morphological characteristics of *L.
pertusa* from different study sites**

Sections will be cut through the skeletons of several different polyps
using a diamond saw. Longitudinal sections will be taken directly across
the calyx, and transverse sections approximately halfway down the
corallite. The sections will be etched with 3N HCl for 10 min, mounted
on stubs, sputter coated with gold and observed under a scanning
electron microscope. This technique has been used to observe skeletal
patterns in *Carophyllia smithii* (Nagelkerken et al. 1997). Sections
will also be mounted on a glass microscope slide with resin and observed
under a light microscope. Growth rates will then be extrapolated using
growth rings. Similar studies are currently underway in the Northern
Gulf of Mexico, where *Lophelia* colonies have been stained '*in situ*'
to calibrate growth rings. This information will assist in the
interpretation of banding patterns observed in other systems since it
will determine the frequency (e.g., annual, biannual) of growth band
deposition. The growth rates and morphology will then be compared
between coral colonies from each region.

**Analysis of population genetic structure of *Lophelia pertusa***

Genetic techniques applied to deep-sea coral samples from various
locations will provide insights into the biology of the habitat forming
corals, mechanisms regulating community structure, and will compliment
other parts of this study through the evolutionary aspect this approach
lends. We will compare phylogeographic structure of *Lophelia* along the
Southeastern U.S. to that in the Gulf of Mexico (using samples gathered
on previous and planned cruises) and the four areas outlined above to
provide perspective on evolutionary units that may exist. We plan to use
microsatellite markers currently under development for U.S. *Lophelia*
to assess fine-scale population genetic structure. Microsatellite use
allows a precise measure of population structure, including an
assessment of gene flow between populations, facilitating identification
of recruitment sources, estimation of effective population size (*Ne*),
and kinship analysis. This approach should be sensitive to detecting
genetic discontinuities between coral bank areas, if they exist, and
will complete sampling efforts from DSCEs from the coast of NC to FL,
and the Gulf of Mexico, allowing for a clear picture of population
structure of *Lophelia* occurring in U.S. EEZ waters. Such information
is critical to effective design of protected areas.

We will obtain tissue/skeleton samples from *Lophelia* from each of the
four study areas described above. Tissue/skeleton samples from other
corals encountered will also be collected to characterize coral
diversity (phylogeny). We will use DMSO and FTA cards for coral tissue
preservation. Digital images of samples will be taken on ship, and
skeletal voucher samples will be saved in either ethanol or dry.

Molecular data will be generated in Tim King's Conservation Genetics lab
(USGS, Leetown Science Center, WV). DNA extractions will be performed
using the PureGene kit (Gentra Systems), and regions of the nuclear
genome containing microsatellite repeats will be amplified via PCR
(markers presently under development in the King lab). Microsatellite
genotyping will take place using an ABI 3100 automated sequencer.
Appropriate computer software for genotype interpretation, phylogenetic,
phylogeographic, and population genetic analyses are available in the
lab.

**Examination of microbial community compositions**

Samples of each coral (living and dead), ambient water (\~4 L per site,
filtered through 0.2 μm final filter, filter retained as actual sample),
and adjacent sediment will be preserved in sterile Tris-EDTA buffer and
frozen at -20^o^C until use. DNA will be extracted from each sample
using commercially available DNA extraction kits. Bacterial DNA will be
amplified using GC-clamped 'universal' 16S rRNA gene primers and the
PCR. Temperature gradient gel electrophoresis (TGGE) will be performed
using the DCode System (BioRad Laboratories) in order to characterize
the bacterial communities associated with each sample. Banding patterns
obtained from the various samples will be compared and bands that are
distinctive to particular environments or samples will be excised,
re-amplified, cloned, and sequenced. Sequences will be used for
phylogenetic analyses and preliminary taxonomic identifications.

**Examination of natural products of selected octocorals**

Specimens of the most abundant species of octocorals collected in the
four regions will be immediately frozen for subsequent analysis of their
marine natural products. Dr. V. Paul will work closely with Dr. S.
Cairns who has experience in the identification of these octocorals.
Shallow water octocorals produce high concentrations of diverse types of
mostly terpenoid secondary metabolites. These have been found to vary
within and between colonies (reviewed by Paul and Puglisi 2004);
however, similar studies of the chemical diversity of deep-water
octocorals have not been conducted. Dr. Paul's research group will
isolate and characterize major metabolites in the octocorals that are
most abundant and examine within- and between-colony variation in the
compounds (by high-performance liquid chromatography (HPLC) or gas
chromatography/mass spectrometry (gc/ms) analytical methods)(e.g.
Slattery et al 2001). Isolation techniques include standard
chromatographic methods including HPLC and structural elucidation will
be conducted primarily by spectroscopic methods (mostly NMR). Results
for the deep-water octocorals will be compared with their closest
shallow water relatives to begin to understand the chemical diversity of
octocorals in these deep-water habitats.

**Education and outreach**

Since 1999, Nova Southeastern University and the School Board of Broward
County (Ft. Lauderdale and surroundings) have maintained a partnership
directed toward improving science education for both teachers and
students. A variety of programs has brought scientists into the
classroom, students, and teachers into the field and laboratory. Because
the expedition will run outside the school year, the 2-person video crew
will record the expedition for subsequent broadcast. We propose to make
the resulting video available on DVD to every school district in the
state of Florida. A separate video, modified in post-production for a
lay audience, will be marketed to public broadcasting outlets.
Supporting materials for teachers beyond those generated aboard ship
will include a CD-ROM modified from Dr. Messing's graduate course
"Deep-water Ecology of the Straits of Florida" (OCMB 6325). We will
select 2-3 teachers to participate in the expedition, document their
cruise experience (eg through interviews with scientists, photographs,
video, onboard lectures, personal notes) and incorporate it to the
science curriculum. One of the proposed participants is Ms. Tracy
Griffin who teaches science at the Space Coast Junior/Senior High School
in Cocoa Beach, and the other two teachers will be selected through the
NSU/Broward County School Board partnership.

Our second outreach effort will be located at the Smithsonian Marine
Ecosystems Exhibit, which is the public outreach arm of the Smithsonian
Marine Station at Fort Pierce. The Exhibit is a low-cost, public
aquarium that presents model ecosystem displays that recreate local
marine and estuarine environments; it is a regular destination for
school groups and a tourist and community attraction. Among the
Exhibit's ecosystem displays is the world's only living *Oculina* coral
reef aquarium and exhibit. We propose to include a new "Expedition
Station" in the exhibit. The station will feature signs that discuss the
objectives of each leg of the deepwater corals cruise and
compares/contrasts other groups and species of deep-sea corals. The
station would also include a mission map that would allow visitors to
track the progress of the research vessel and an expedition board that
will display daily dispatches from the science crew provided by the NOAA
Ocean Exploration web site. After the cruise, the update board would be
replaced by an expedition summary, and a push-button interactive that
highlights the distribution of deep-sea coral ecosystems would be added.
We are proposing three 2' x 1' signs hanging in a column, topped with a
2' x 4' title graphic. The top sign would have details on the cruise and
its objectives and an expedition map. The second sign would have notes
on the scientists and space for the daily dispatches. The bottom sign
would have information about groups/species of deep-sea corals. There
are two possibilities for interactive exhibits. The first is a simple
way for visitors to locate the research vessel\'s location based on the
longitude and latitude from the daily dispatches. After the cruise is
finished, this could be used as an interactive for school group
scavenger hunts in conjunction with the second interactive. The second
idea is a light board that would reflect the location of different
deep-sea coral species/communities found during the cruise. The cost
would include the design and fabrication of three signs, one title
graphic, the wiring for the interactive plus installation. This exhibit
could provide a public outreach effort for the entire duration of the
deep-sea corals expedition and beyond, not our proposed leg alone.

# Key Personnel

**Dr. Sandra Brooke**

Dr. Brooke will be the cruise PI and will be responsible for the
coordination of cruise logistics and management of the grant funds.
After the cruise, assist in the analysis of videotapes for habitat
characterization, process samples for determination of reproductive
status, measurement of growth rates and description of coral morphology.
She will also assist in coordination of outreach efforts and compilation
of data for reports and publications.

## 

## Mr. John Reed 

Mr. Reed is a Co-PI on the proposal and will select potential dive
target sites during the cruise, based on previous knowledge and dives,
and will assist with habitat mapping and characterization, including
analysis of video transects for benthic habitat, and identification of
sessile macrofauna including corals, octocorals and sponges.

## 

## Dr. Charles Messing

Dr. Messing is also a Co-PI on the proposal and will be responsible for
documentation of biozonation patterns of both sessile and vagile benthic
fauna, particularly relative to bathymetry and hydrodynamic regime, and
will contribute to identification of invertebrates. He will also play a
significant role in coordinating educational outreach and video
production.

**Dr. R. Grant Gilmore**

Grant Gilmore will classify and quantify all fish species documented
during video transects or captured with collection apparatus. He will be
responsible for determining relative habitat association of the species
documented.

## 

## Dr. Cheryl Morrison

Dr. Morrison will collect tissue samples of corals for population
genetics and phylogeny work to be completed at the USGS Leetown Science
Center. *Lophelia* samples will be incorporated into an ongoing
population genetic study covering the Southeastern U.S. and the Gulf of
Mexico.

**Dr. Julie Olson**

Dr. Olson will examine and characterize the microbial communities
associated with the dominant coral species, and determine whether
symbiotic bacteria exist within coral tissue.

### 

### Dr. Valerie Paul

Dr. Paul will investigate the ecology and chemical ecology of associated
fauna, inventory samples for the Smithsonian museum, and coordinate
public outreach through Smithsonian Marine Ecosystems Exhibit in Fort
Pierce, Florida.

**Dr Stephen Cairns**

Dr. Cairns of the Smithsonian Institution National Museum of Natural
History will be responsible for identification of coral species
(scleractinians, hydrocorals and gorgonians), and for deposition of
voucher specimens in the National Collections.

**Dr. J. Harasewych**

Dr Jerry Harasewych, also of the NMNH will identify mollusks, and
deposit voucher specimens in the National Collections. If a particular
molluskan genus or species is common and can be readily collected, Dr
Harasewych will investigate its population genetic structure, in order
to compare the biogeography of deep and shallow water mollusks from
around the Florida peninsula.

**Dr. William Schroeder**

Dr. Schroeder will be responsible for multi-scale geological
characterization of the different regions.

**[Proposed schedule]{.smallcaps}**

Day 1-5 Transit to Region 4 on the southwest Florida shelf and conduct
Multibeam (or mapping surveys and ROV dives.

Day 6-8 Transit to Region 3, Pourtalès Terrace sites (\~240 nm) and
conduct multibeam mapping surveys and ROV dives

Day 9-12 Transit to Region 2, Miami Terrace sites (\~60 nm) and conduct
multibeam mapping surveys and ROV dives

Day 13-17 Transit to Region 1, Florida Lophelia sites (\~120 nm), and
conduct multibeam mapping surveys and ROV dives.

**[Relevance to the goal of NOAA\'s Ocean Exploration
Program]{.smallcaps}**

Information from the literature and from past expeditions provides
strong evidence that each of the proposed study regions supports
well-developed coral habitats. The proposed research will expand our
limited understanding of these habitats, providing detailed descriptions
of coral distribution and abundance, and coral-associated fauna
(including fish) at representative sites within each region. This
research will significantly contribute to four of the preferred themes
designated under OE's Priority Objectives: mapping of deep-water coral
systems, characterization of benthic and pelagic habitats and
ecosystems, species inventories, and seafloor mapping. This information
can be used by the SFMC to determine appropriate protection measures and
by scientists as a basis for future hypothesis driven research.
Additionally, the outreach components will significantly and
substantially expand the understanding of these poorly known
environments by Florida's students and science teachers as well as the
public.

**[Literature Cited]{.smallcaps}**

Ballard R, Uchupi E (1971) Geological observations of the Miami Terrace
from the submersible Ben Franklin. Marine Tech Society Journal 5:43-48

Bogle MA (1975) A review and preliminary revision of the Aglaopheniinae
(Hydroida: Plumulariidae) of the tropical western Atlantic. Unpublished
M.S. Thesis, University of Miami

Brooke SD (2002) Reproductive ecology of a deep-water scleractinian
coral, *Oculina varicosa* from the South East Florida Shelf.
Dissertation, Southampton Oceanography Centre, Southampton

Brooke SD, Young CM (2003) Reproductive ecology of a deep-water
scleractinian coral, *Oculina varicosa* from the South East Florida
Shelf. Cont Shelf Research 23:847-858

Cairns SD (1979) The deep-water scleractinia of the Caribbean Sea and
adjacent waters. In Hummelinck, PW & LJ Van der Steen (eds) Studies of
the Fauna of Curacao and other Caribbean Islands 180, 341 pp

Cairns SD (l986) A revision of the Northwest Atlantic Stylasteridae
(Coelenterata: Hydrozoa). Smith Cont Zool 4l8, l3l pp, 53 pls, 25 maps

Cairns SD, Chapman RE  (2002)  Biogeographic affinities of the North
Atlantic deep-water Scleractinia. In Willison, JHM et al. (eds) Proc.
First Internat Symp Deep-Sea Corals. Ecology Action Centre and Nova
Scotia Museum, Halifax, pp 30-57

 

Freiwald A, Henrich R, Pätzold J (1997) Anatomy of a deep-water coral
reef mound from Stjernsund, west Finnmark, northern Norway. Soc sedim
Geol, SEPM spec. Pub 56: 141-161

Gilmore RG, Jones RS (1992) Color variation and associated behavior in
the epinepheline groupers, *Mycteroperca* *microlepis* (Goode and Bean)
and *M*. *phenax* Jordan and Swain. Bull Mar Sci 51(1): 83-103

Gomberg, D  (1976) Geology of the Pourtales Terrace, Straits of Florida.
Ph.D. Dissertation, Univ. Miami, Fl

Halpern JA (1970) Goniasteridae (Echinodermata: Asteroidea) of the
Straits of Florida. Bull Mar Sci 20(1):193-286

Holthuis LB (1971) The Atlantic shrimps of the deep-sea genus
*Glyphocrangon* A. Milne

Edwards, 1881. Bull Mar Sci 21(1):267-373

Holthuis LB (1974) The lobsters of the superfamily Nephropidea of the
Atlantic Ocean (Crustacea: Decapoda). Bull Mar Sci 24(4):723-871

Jordan GF, Stewart HB (1961) Submarine topography of the western Straits
of Florida. Geological Society of America Bulletin 72(7):1051-1058

Jordan GF, Malloy RM, Kofoed JW (1964) Bathymetry and geology of
Pourtales Terrace, Florida. Marine Geology 1(3):259-287

Kofoed JW, Malloy RM (1965) Bathymetry of the Miami Terrace.
Southeastern Geology 6(3):159-165

Koslow JA, Gowlett-Jones K (1998) The Seamount Fauna off Southern
Tasmania: Benthic Communities, Their Conservation and Impacts of
Trawling. Final Report to Environment Australia and Fisheries Research
Development Coorporation. Australia, 104 pp

Land L, Paull C (2000) Submarine karst belt rimming the continental
slope in the Straits of Florida.  Geo-Marine Letters 20: 123-132

Malloy RJ, Hurley R (1970) Geomorphology and geologic structure: Straits
of Florida. Geological Society of America Bulletin 81: 1947-1972 \[map
of Florida Straits\]

Messing CG, Neumann AC, Lang JC (1990) Biozonation of deep-water
lithoherms and associated hardgrounds in the northeastern Straits of
Florida. Palaois 5(1): 15-33

Meyer DL, Messing CG, Macurda Jr DB (1978) Zoogeography of tropical
western Atlantic Crinoidea. Bull Mar Sci 28:412-441

Nagelkerken I, Van der Velde G, Van Avessaath PH (1997) A description of
the skeletal development pattern of the temperate coral *Carophyllia
smithii* based on internal growth lines. J Mar Biol Ass UK 77: 375-387

Neumann AC, Ball MM (1970) Submersible observations in the Straits of
Florida: geology and bottom currents. Geol Soc Am Bull 81: 2861-2874

Neumann AC, Kofoed JW, Keller GH (1977) Lithoherms in the Straits of
Florida. Geology 5: 4-10

Paul VJ, Puglisi MP (2004) Chemical mediation of interactions among
marine organisms. Nat Prod Rep 21: 189-209

Paull CK, Neumann A, am Ende B, Ussler W, Rodriguez N (2000) Lithoherms
on the Florida-Hatteras slope.  Marine Geology 166: 83-101

Reed JK (1980) Distribution and structure of deep-water *Oculina
varicosa* coral reefs off central eastern Florida. Bull Mar Sci 30:
667-677

Reed JK (1981) *In situ* growth rates of the scleractinian coral
*Oculina varicosa* occurring with zooxanthellae on 6-m reefs and without
on 80-m banks. In Dogma Jr IJ (ed) Proc 4th Inter Coral Reef Symp 2:
201-206

Reed JK (1983) Nearshore and shelf-edge *Oculina* coral reefs: the
effects of upwelling on coral growth and on the associated faunal
communities. NOAA Symp Ser Undersea Res 1: 119-124

Reed JK (1992) Submersible studies of deep-water *Oculina* and
*Lophelia* coral banks off

southeastern U.S.A. In Cahoon LB (ed) Proc Am Acad Underwater Sci 12^th^
Ann Scien Diving Symp:143- 151

Reed JK, Wright A (2004) Final cruise report.  Submersible and scuba
collections on deep-water reefs off the east coast of Florida, including
the Northern and Southern Straits of Florida and Florida Keys National
Marine Sanctuary for biomedical and biodiversity research of the benthic

communities with emphasis on the Porifera and Gorgonacea, May 20- June
2, 2004.  Conducted by the Center of Excellence, HBOI and FAU, pp 54

Reed JK, Pomponi S, Wright A, Weaver D, Paull C  (2004,
submitted)  Deep-water sinkholes and lithoherms of South Florida and
Pourtales Terrace- Habitat and Biology

Reed JK (2004, in prep) Habitat characterization and comparison of five
deep-water coral reefs: Blake Plateau, Straits of Florida, and the Gulf
of Mexico

Reed K (2002) Comparison of deep water coral reefs and Lithoherms off
southeastern USA. Hydrobiologia 471(1-3):57-69

Slattery M, Starmer J, Paul VJ (2001)  Temporal and spatial variation in
defensive metabolites of the tropical Pacific soft corals Sinularia
maxima and S. polydactyla.  Mar Biol 138: 1183-1193

Stetson TR, Squires DF, Pratt RM (1962) Coral banks occurring in deep
water on the Blake Plateau. Am Mus Nov 2114: 1-39

Teichert C (1958) Cold- and deep-water coral banks. Bull Am Ass petrol
Geol 42: 1064-1082

Uchupi E (1966) Shallow structure of the Straits of Florida. Science
153: 529-531

Uchupi E (1969) Morphology of the continental margin southeastern
Florida. Southeastern Geology 11: 129-134

Uchupi E, Emery K  (1967) Structure of continental margin off Atlantic
coast of United States.  Amer Assoc of Petroleum Geologists Bull 51:
223-234

Waller RG, Tyler PA (in Press) The reproductive biology of two deep-sea,
reef-building scleractinians from the NE Atlantic Ocean. Coral Reefs

Wilson JB (1979a) The distribution of the coral *Lophelia pertusa* (L)
\[*L. prolifera* (Pallas)\] in the northeast Atlantic. J Mar Biol Ass UK
59: 149-164

Wilson JB (1979b) "Patch" development of the deep-water coral *Lophelia
pertusa* (L.) on Rockall Bank. J Mar Biol Ass UK 59: 165-177

Wise JB, Harasewych MG, Dillon Jr RT  (2004) Population divergence in
sinistral whelks of North America, with special reference to the east
Florida ecotone. Mar Biol 145(4): (available online - hard copy in
press)