68 Marine Technology Society Journal AUTHORS James Lindholm Stellwagen Bank National Marine Sanctuary; Current address: Pfleger Institute of Environmental Research Sarah Fangman Channel Islands National Marine Sanctuary Les Kaufman Boston University Marine Program Steven Miller National Undersea Research Center, University of North Carolina at Wilmington TECHNICAL NOTE In Situ Tagging and Tracking of Coral Reef Fishes from the Aquarius Undersea Laboratory ABSTRACT We surgically implanted coded-acoustic transmitters in a total of 46 coral reef fish during a saturation mission to the Aquarius Undersea Laboratory in August 2002. Aquarius is located within the Conch Reef Research Only Area, a no-take marine re- serve in the northern Florida Keys National Marine Sanctuary. Over the course of 10 days, with daily bottom times of 7 hrs, saturation diving operations allowed us to col- lect, surgically tag, release, and subsequently track fishes entirely in situ. Fish were collected using baited traps deployed adjacent to the reef as well as nets manipulated on the bottom by divers. Surgical implantation of acoustic transmitters was conducted at a mobile surgical station that was moved to different sites across the reef. Each fish was revived from anesthetic and released as divers swam the fish about the reef. Short- term tracking of tagged fish was conducted by saturation divers, while long-term fish movement was recorded by a series of acoustic receivers deployed on the seafloor. Though not designed as an explicit comparison with surface tagging operations, the benefits of working entirely in situ were apparent. T INTRODUCTION he use of acoustic telemetry to track the movements of marine fishes is now a com- monly employed method (see other papers this issue), producing data on fish home ranges (Zeller, 1997; Bolden, 2001; Simpfendorfer et al., 2002), habitat-specific movement (Lindholm and Auster, 2003; Lowe et al., 2003; Cote et al., 2004), and movement relative to the boundaries of marine protected areas, or MPAs (Zeller and Russ, 1998; Meyer et al., 2000; O’Dor et al., 2001; Starr et al., 2001; Lowe et al., 2003). While the specific approaches to the use of acoustic telemetry vary widely depend- ing on the species targeted for study and the habitat in which the targeted species occurs, all projects share four common elements: 1) the collection of fishes, 2) the tagging of fishes, 3) the release of tagged fishes, and 4) the tracking of tagged fishes. Also common to most studies is the conduct of each of these elements from the surface, where the extraction of fishes from their natural envi- ronment can create difficulties. Fishes are often collected from the sur- face via baited traps, long-lines, trolling and traditional angling. Fish brought to the sur- face for tagging can experience barotrauma, or pressure-related stress. This is particularly true with deepwater fishes that have air blad- ders (Starr et al., 2000). Thermal shock can also result from bringing a fish to the sur- face and then aboard a surface vessel (Kelsch and Shields, 1996). Each of these stressors can kill a fish outright or have sub-lethal ef- fects that increase the fishes’ vulnerability to other stressors and may preclude a fish from being tagged. Telemetry projects on fishes typically involve either the attachment of an acoustic transmitter externally (e.g., Bradbury et al., 1995; Lindholm and Auster, 2003; Cartamil and Lowe, 2004), intragastric insertion of the transmitter down the pharynx into the stomach (Bridger and Booth, 2003), or sur- gical implantation of the transmitter inside the peritoneal cavity of a fish (e.g., Zeller, 1997; Starr et al., 2000; Bolden, 2001; pa- pers in this volume). Where surgical implan- tation is used, incisions can be closed with sutures (e.g., Thoreau and Baras, 1997), sur- gical staples (e.g., Mortensen, 1990) or an adhesive (e.g., Nemetz and MacMillan, 1988). Implantation is normally conducted on a padded, v-shaped surgical board (Win- ter, 1996) and may involve fresh flowing seawater over a fish’s gills, or covering the fish with a damp towel. Each approach in- volves a series of trade-offs that will vary depending on the species selected for tag- ging, though most approaches involve ex- posure of the fish to air and sunlight for some period of time during surgery. Following some period of observation in a live-well on board a research vessel, tagged fishes are either released directly over the side of the boat or are lowered to the seafloor in some form of a release device (e.g., Starr et al., 2000; Lindholm and Auster, 2003). Though the use of a release device can increase researcher confidence that the fish has indeed returned to the sea- floor or at the depth from which it was collected initially, both forms of release in- volve a high measure of uncertainty. The fate of these fish post-release is difficult to determine, i.e., did the transmitter mal- function, was the tagged fish consumed by a predator, is the fish dead and lying on the seafloor? This problem is lessened when data are collected indicating the tagged fish is moving. However, where the post-release data are limited or non-existent, it is often impossible to know the fate of the fish with- out in situ observation.
Transcript
68 Marine Technology Society Journal
A U T H O R SJames LindholmStellwagen Bank National Marine Sanctuary;Current address: Pfleger Institute ofEnvironmental Research
Sarah FangmanChannel Islands National Marine Sanctuary
Les KaufmanBoston University Marine Program
Steven MillerNational Undersea Research Center,University of North Carolina at Wilmington
T E C H N I C A L N O T E
In Situ Tagging and Tracking of Coral Reef Fishesfrom the Aquarius Undersea Laboratory
A B S T R A C TWe surgically implanted coded-acoustic transmitters in a total of 46 coral reef fish
during a saturation mission to the Aquarius Undersea Laboratory in August 2002.Aquarius is located within the Conch Reef Research Only Area, a no-take marine re-serve in the northern Florida Keys National Marine Sanctuary. Over the course of 10days, with daily bottom times of 7 hrs, saturation diving operations allowed us to col-lect, surgically tag, release, and subsequently track fishes entirely in situ. Fish werecollected using baited traps deployed adjacent to the reef as well as nets manipulatedon the bottom by divers. Surgical implantation of acoustic transmitters was conductedat a mobile surgical station that was moved to different sites across the reef. Each fishwas revived from anesthetic and released as divers swam the fish about the reef. Short-term tracking of tagged fish was conducted by saturation divers, while long-term fishmovement was recorded by a series of acoustic receivers deployed on the seafloor.Though not designed as an explicit comparison with surface tagging operations, thebenefits of working entirely in situ were apparent.
TI N T R O D U C T I O N
he use of acoustic telemetry to track themovements of marine fishes is now a com-monly employed method (see other papersthis issue), producing data on fish homeranges (Zeller, 1997; Bolden, 2001;Simpfendorfer et al., 2002), habitat-specificmovement (Lindholm and Auster, 2003;Lowe et al., 2003; Cote et al., 2004), andmovement relative to the boundaries ofmarine protected areas, or MPAs (Zeller andRuss, 1998; Meyer et al., 2000; O’Dor etal., 2001; Starr et al., 2001; Lowe et al.,2003). While the specific approaches to theuse of acoustic telemetry vary widely depend-ing on the species targeted for study and thehabitat in which the targeted species occurs,all projects share four common elements: 1)the collection of fishes, 2) the tagging offishes, 3) the release of tagged fishes, and 4)the tracking of tagged fishes. Also commonto most studies is the conduct of each ofthese elements from the surface, where theextraction of fishes from their natural envi-ronment can create difficulties.
Fishes are often collected from the sur-face via baited traps, long-lines, trolling andtraditional angling. Fish brought to the sur-face for tagging can experience barotrauma,or pressure-related stress. This is particularly
true with deepwater fishes that have air blad-ders (Starr et al., 2000). Thermal shock canalso result from bringing a fish to the sur-face and then aboard a surface vessel (Kelschand Shields, 1996). Each of these stressorscan kill a fish outright or have sub-lethal ef-fects that increase the fishes’ vulnerability toother stressors and may preclude a fish frombeing tagged.
Telemetry projects on fishes typicallyinvolve either the attachment of an acoustictransmitter externally (e.g., Bradbury et al.,1995; Lindholm and Auster, 2003; Cartamiland Lowe, 2004), intragastric insertion ofthe transmitter down the pharynx into thestomach (Bridger and Booth, 2003), or sur-gical implantation of the transmitter insidethe peritoneal cavity of a fish (e.g., Zeller,1997; Starr et al., 2000; Bolden, 2001; pa-pers in this volume). Where surgical implan-tation is used, incisions can be closed withsutures (e.g., Thoreau and Baras, 1997), sur-gical staples (e.g., Mortensen, 1990) or anadhesive (e.g., Nemetz and MacMillan,1988). Implantation is normally conductedon a padded, v-shaped surgical board (Win-ter, 1996) and may involve fresh flowingseawater over a fish’s gills, or covering the
fish with a damp towel. Each approach in-volves a series of trade-offs that will varydepending on the species selected for tag-ging, though most approaches involve ex-posure of the fish to air and sunlight for someperiod of time during surgery.
Following some period of observationin a live-well on board a research vessel,tagged fishes are either released directly overthe side of the boat or are lowered to theseafloor in some form of a release device(e.g., Starr et al., 2000; Lindholm andAuster, 2003). Though the use of a releasedevice can increase researcher confidencethat the fish has indeed returned to the sea-floor or at the depth from which it wascollected initially, both forms of release in-volve a high measure of uncertainty. Thefate of these fish post-release is difficult todetermine, i.e., did the transmitter mal-function, was the tagged fish consumed bya predator, is the fish dead and lying on theseafloor? This problem is lessened whendata are collected indicating the tagged fishis moving. However, where the post-releasedata are limited or non-existent, it is oftenimpossible to know the fate of the fish with-out in situ observation.
69Spring 2005 Volume 39, Number 1
Tracking of tagged fishes is often con-ducted manually from the surface (e.g.,Matthews, 1992; Zeller, 1997; Cartamil andLowe, 2004), recorded by acoustic receiversdeployed on seafloor (e.g., Arendt et al.,2001; Starr et al., 2001; Lindholm andAuster, 2003), or a combination of the twoapproaches (e.g., Simpfendorfer et al., 2002;Lowe et al., 2003; Parsons et al., 2003). Theapproach used to study the movement oftagged fishes will depend heavily on the spe-cies tagged, the region in which it was tagged,and the objectives of the study.
Obviously, the success of acoustic telem-etry approaches to tracking marine fishes iscontingent on tagging fish in a manner thatdoes not negatively affect their behavior andphysiology (Smolowitz and Wiley, 1998;Bridger and Booth, 2003). Thus any ap-proach that minimizes the potential stres-sors at each step of the process is preferable.In this paper we describe a study using acous-tic telemetry to track movements of coralreef fishes in which all aspects of the projectwere conducted in situ from the AquariusUndersea Laboratory (Figure 1).
Aquarius is currently located within theConch Reef Research Only Area, a no-takereserve designated by the Florida Keys Na-tional Marine Sanctuary. It provided a plat-form for a 10-day saturation diving missionin August 2002. A total of 70 hrs of bottomtime (~ 7 hr day-1) provided saturation diverswith sufficient time to tag and subsequentlyobserve 46 fish (Table 1), including blackgrouper (Mycteroperca bonaci), yellowtailsnapper (Ocyurus chrysurus), princessparrotfish (Scarus taeniopterus), blueparrotfish (Scarus coeruleus), and hogfish(Lachnolaimus maximus). We describe someof the lessons learned during the mission.
Fish CollectionCollection of fishes in situ provided us a
high degree of selectivity in the fish we choseto tag. The majority of fishes were caught inbaited traps (1.5 m2) deployed at two loca-tions on sand flats immediately adjacent toConch Reef during daylight hours (Figure 2).Both locations were sited within 300 m ofAquarius to ensure easy access to saturation
divers. Each trap was visited by saturationdivers at the beginning of a dive, with subse-quent visits depending upon the number andspecies of fishes caught by the trap. In casesin which multiple fishes within a trap wereselected for tagging, the opening to the trapwas blocked and each fish was tagged sequen-tially until the trap was empty. The trap wouldthen be baited again and re-deployed.
FIGURE 1The Aquarius undersea laboratory (bow and starboard view) at Conch Reef in the northern Florida Keys NationalMarine Sanctuary. Photo credit: James Lindholm
TABLE 1List of coral reef fish species tagged at Conch Reefduring the August 2002 Aquarius mission.
IndividualsSpecies Tagged
Yellowtail snapper 14Mangrove snapper 2*
Black grouper 1 (2*)Scamp grouper 1
Hogfish 10
Blue parrotfish 8Princess parrotfish
Initial Phase 4Terminal Phase 8
*These fish were tagged from the surfaceimmediately prior to the start of the mission.
Yellowtail snapper, blue and princessparrotfish, as well as black grouper were eachcaught in traps during the saturation mis-sion. The multiple yellowtail snapper cap-tured in traps appeared to be attracted tothe bait (punctured cans of cat food), whilethe single black grouper caught in a trapappeared to be attracted more to the varietyof fishes contained within the trap. Bothspecies of parrotfishes appeared to enter thetraps out of curiosity, often following con-specifics into the trap. Other species thatwere captured (but not tagged) includedchub (Kyphosus spp.), French grunt(Haemulon flavolineatum), reef butterflyfish(Chaetodon sedentarius), gray angelfish(Pomacanthus arcuatus), blue tang(Acanthurus coeruleus), doctorfish(Acanthurus chirurgus), and green moray(Gymnothorax funebris).
Our ability to capture fish in greaternumbers may have been reduced by the pres-ence of large predators immediately adjacent
70 Marine Technology Society Journal
to the traps. A nurse shark (Ginglymostomacirratum) was commonly observed swim-ming near one of the traps, and on one oc-casion was found resting on top of a trap.Though there were no observed acts of pre-dation within the trap, on two occasions agreen moray was found in the trap stationedadjacent to Aquarius. In each case the trapwas otherwise devoid of fish, though noobvious distensions of the eel’s stomach wereobserved so it was not clear whether trappedfish had been consumed. During the No-vember 2001 Aquarius mission, a greenmoray when disturbed egested a black grou-per similar to one that had previously beenseen in the trap (Lindholm et al., unpub-lished observations).
In addition to the selective tagging offishes from the traps, the extensive bottomtimes provided by saturation diving also al-lowed us to selectively target individual fishfor capture using alternative approaches. Astandard beach seine net was used to corralfish that were foraging on the sand flat adja-cent to Aquarius. In each case, one end ofthe net was anchored to the seafloor while
the opposing end was handled by a diver. Asecond diver would gradually herd fish to-ward the net as the other diver closed thenet (as per Eristhee et al., 2001). Using thisapproach, a total of 5 hogfish (which gener-ally avoided the traps) and a blue parrotfishwere captured. Though time consuming, theuse of the net allowed us to select particularfish for capture and tagging with some suc-cess. An additional 3 hogfish and a scampgrouper were herded directly into a diver’smesh bag by one of the Aquarius technicianswhile conducting maintenance on the lab’sexternal surfaces. The hogfish were attractedto the fouling being chipped off the Aquariussupport columns by the technician.
The selectivity provided by each of thesethree approaches (trap, net, bag) would ei-ther not have been possible with most sur-face tagging operations, or would only bepossible to a limited extent as surface diverswould be constrained to shorter bottomtimes. By effectively “pre-screening” thefishes prior to surgery (Starr et al., 2000),we were able minimize the collection of non-targeted species, a major consideration when
working within a fully-protected marine re-serve. While collection of non-targeted fisheswas common with the traps, all were releasedshortly after capture and with the exceptionof the possible consumption by the greenmorays, no by-catch mortality was observed.
Fish TaggingThe conduct of surgical tag insertion in
situ is not new (Starr et al., 2000), but is lesscommon than tagging from the surface. Inthe present study all fishes were surgicallytagged with acoustic transmitters at the samelocation and depth where they were col-lected. Protocols for surgery were developedduring the November 2001 Aquarius mis-sion (Stone, 2003). Results from previoustagging efforts at Conch Reef indicated thatthe number of signal detections for yellow-tail snapper and black grouper was signifi-cantly higher for those fish with intra-peri-toneal transmitters when compared toexternally attached transmitters (Lindholmet al., unpublished observations). We hy-pothesized that the externally attached trans-mitters, particularly for yellowtail snapper,served to attract predators and disrupt swim-ming ability.
Once a captured fish was selected forsurgical implantation of a transmitter, it wasisolated and removed from the trap using adiver’s mesh bag. While in the mesh bag,each fish was anesthetized using seawater-buffered MS-222 solution (100 mg l-1;Summerfelt and Smith, 1990) contained ina 120 ml syringe. Administering anestheticat the surface often involves submerging afish in an anesthetic bath until complete stu-por is achieved. This approach has provensuccessful, but may expose fish to higherdosages of anesthetic than are necessary.Further, the effective and toxic doses of MS-222 are similar, particularly in warmer wa-ters (Bridger and Booth, 2003). Using thesyringe, the anesthetic was introduced di-rectly into the mouth of the fish while stillin the mesh bag. Once signs of stupor wereevident, fish were removed from the meshbag and additional anesthetic was providedif necessary. In this way each fish was ex-posed only to the minimum amount of an-
FIGURE 2Map of VR2 acoustic receiver array at Conch Reef, including the estimated range of detection for each receiver,the boundary of the Conch Reef Research Only Area, the location of the Aquarius undersea laboratory, and thelocations where fish were collected and released. Five meter isobaths are provided. The complete receiver array(inset) is shown extending from Alligator Reef to Carysfort Reef in the northern Florida Keys.
71Spring 2005 Volume 39, Number 1
esthetic necessary to attain complete stupor.Stage IV anesthesia was achieved in 1-3 min-utes depending on species and fish size.
Following anesthesia, each fish was re-moved from the mesh bag and placed in amesh sling designed to support the fish dur-ing surgery (Figure 3). The sling was sup-ported by a PVC frame and was easily trans-ported by divers to different locations on thereef. A 25 mm incision was made just off theventral line on the left side of the fish, for-ward of the vent. Each fish was tagged with aV8SC-1H (69 kHz) coded-acoustic transmit-ter (VEMCO, Ltd., Shad Bay, Nova Scotia).Each transmitter, measuring 3.3 cm in lengthand weighing 3.5 gm in seawater, randomlyproduced a unique ID code at intervals be-tween 60 –180 seconds. Each transmitter wascoated with triple-antibiotic ointment andinserted into the peritoneal cavity. The inci-sion was closed using 1 to 3 black monofila-ment sutures (5-0 Ethilon) and coated withtriple-antibiotic ointment. Total length (TL)was measured for each fish and an externalflag tag was inserted into the musculature atthe base of the first dorsal fin.
The release of tagged fish in situ follow-ing surgery provided considerable advantages
over a release from the surface. A diver swameach tagged fish along the seafloor to flushits gills with “fresh” seawater until it com-pletely revived. Each fish required 2-5 min-utes to revive completely depending on thesize and species. Once fully revived, each fishwas released on the seafloor and was observedfor 5 minutes, or until the fish swam out ofvisual range. All 46 tagged fishes were ob-served to recover from the surgical proce-dure and resume normal swimming behav-ior following release. The single mortalitywe observed occurred when a tagged yellow-tail snapper was consumed by large blackgrouper shortly after release by a diver. Thestrike occurred in the water column as theyellowtail snapper swam toward a large ag-gregation of conspecifics.
Observation and Trackingof Tagged Fishes
Another distinct advantage of conduct-ing in situ operations was our ability to ob-serve and evaluate the condition of mosttagged fishes following release. A diver-heldVUR96 receiver (VEMCO Ltd., Shad Bay,Nova Scotia) was used to track a subset ofall tagged fishes. In addition to the coded
V8SC-1H transmitters, a total of 8 fisheswere also tagged with V8SC continuoustransmitters, including 2 black grouper, 4blue parrotfish, and 2 mangrove snapper(Lutjanus griseus). The 2 black grouper and2 mangrove snapper were tagged from thesurface immediately prior to the beginningof the mission. These transmitters were simi-larly dimensioned to the coded transmitters,but transmitted a signal each second. Usingthe diver-held receiver, each of the 8 fish waslocated over the course of the 10-day mis-sion on multiple occasions. Though satura-tion diving rules limited the distance diverswere permitted to venture away fromAquarius, fish located using the diver-heldreceiver were observed for periods of up to15 minutes at a time or until the fish swamout of visual range.
Additional tagged fishes were identifiedby their external tags and were observed forcomparable periods of time (Figure 4).Though the tag ID numbers were not pos-sible to read without re-capturing the fish,the combination of fish size, location onthe reef, and unique markings or scars al-lowed us to identify particular fish withhigh confidence. Each of the 10 taggedhogfish were observed in this manner, aswere each of the 8 blue parrotfish, severalof the princess parrotfish (both initial andterminal phase fish), and two of the 13tagged yellowtail snapper. In each case, theprimary observation was to confirm thefishes were alive and swimming freely. Ad-ditional observations included post-taggingfeeding behavior and microhabitat utiliza-tion. The behavior of tagged fishes wascompared to untagged control fish fromeach species to determine whether tagginghad altered fish behavior (Lindholm et al.,unpublished observations).
Long-term patterns in the movement oftagged fishes (up to 11 months) were re-corded by three, omni-directional, single-channel (69 kHz) VR2 acoustic receivers(VEMCO, Ltd., Shad Bay, Nova Scotia)deployed at Conch Reef (Figure 2) at 25 m(Site 1), 20 m (Site 2), and 25 m (Site 3),respectively. Each VR2 receiver operated incontinuous receiving mode, recording thepresence/absence of tagged fish within a
FIGURE 3Saturation divers conduct surgical implantation of an acoustic transmitter in a coral reef fish using a portablePVC surgical station. Photo credit: Rick Riera-Gomez/RSMAS
72 Marine Technology Society Journal
range of detection of approximately 300 m(based on preliminary testing). Receiverswere placed by divers from the surface im-mediately prior to the Aquarius mission.They were mounted on bars drilled into thereef that maintained the receiver heads ori-ented toward the surface at a height of 1 mabove the seafloor.
Placement of the receivers inside theConch Reef Research Only Area providedprotection from disturbance by recre-ational diving and fishing activity, bothof which are excluded from the area sur-rounding Aquarius. An additional 24 VR2receivers were placed along the reef tractfrom Alligator Reef in the south toCarysfort Reef in the north, encompass-ing 40 km (Figure 2 inset). Many of thesereceivers were also located within Sanctu-ary Preservation Areas (another type of no-take reserve within the Florida Keys Na-tional Marine Sanctuary). They were sitedto capture any movement of tagged fishesaway from Conch Reef.
Data were collected by the 3 VR2 re-ceivers deployed at Conch Reef for 49 ofthe 50 tagged fish during the first monthfollowing release, with data for selected in-dividual fish collected for up to 11 months.No data were collected on the yellowtail
consumed shortly after release. Further, nodata were collected at receivers to the northor south of Conch Reef (Lindholm et al.,unpublished information). All receivers weredeployed from August 2002 through July2003, three weeks following the last recordof a tagged fish.
ConclusionsThe conduct of a fish tagging project en-
tirely in situ from Aquarius proved highly suc-cessful. Though we did not design this studywith an explicit surface tagging componentfor a direct comparison, the experience wasinstructive. Most importantly, tagged fishesnever left the depths at which they occurrednaturally. The capture and surgical tagging ofcoral reef fishes without bringing them to thesurface minimized stress on the selected fishes,and minimized the capture of non-targetedspecies. The sub-surface release of fish bydivers and the subsequent observation of post-surgery fish behavior over the course of 10days provided further evidence of the ben-efits of working in situ. This provided a levelof confidence in the resulting movement datathat is not often possible with operations con-ducted from the surface.
FIGURE 4A tagged hogfish forages adjacent to Aquarius one day following surgical implantation of an acoustic transmit-ter. The surgical incision is visible just off the ventral line, forward of the anal fin, while the external flag tag isclearly visible at the base of the dorsal fin. Photo credit: Rick Riera-Gomez/RSMAS
Obviously, saturation diving is not pos-sible under all circumstances. Aquarius is cur-rently the only undersea laboratory in theworld dedicated to scientific research (Millerand Cooper, 2001). However, though ourability to conduct the project entirely in situderives largely from the extensive bottom timeprovided by saturation diving, saturation neednot occur to successfully conduct operationsunderwater. For instance, Starr et al. (2000)developed an approach for tagging deepwaterrockfishes using standard SCUBA to tag fishat a surgical station deployed 20 m below asurface vessel. Fish caught on hook-and-linewere reeled up to divers waiting at the sta-tion, and were returned to the seafloor post-tagging in a release device without ever reach-ing the surface.
Ultimately, the object of any acoustic te-lemetry project is to produce data in whichthe investigators have high confidence that therecorded patterns in movement are indeed re-flective of living fish behaving similarly to un-tagged conspecifics in the same area. To theextent that any component of a telemetryproject can be conducted in situ, our experi-ence suggests it will be worth the effort.
AcknowledgementsWe would like to thank Kim Benson
and William Ojwang for assistance fromthe surface throughout the mission. Wewould also like to thank Mike Feeley andRick Rierra-Gomez from RSMAS, ThorDunmire of NURC-UNCW for key as-sistance in catching fish, and the rest ofthe NURC-UNCW staff for assistance atall stages of the project. Support for theproject was provided by NOAA’s NationalMarine Sanctuary Program, the NationalMarine Fisheries Service, and privategrants to Steven Miller. The views ex-pressed herein represent those of the au-thors and do not necessarily reflect theviews of NOAA or any of its sub-agen-cies. This work was conducted under per-mit by the Florida Keys National MarineSanctuary (FKNMS-2002-053) and wasapproved by the Institutional Animal Careand Use Committee at Boston University(Protocol # 02-020).
