DIVING FOR SCIENCE… 2003

Proceedings of the American Academy of Underwater Sciences

22nd Annual Scientific Diving Symposium

March 14 – 15, 2003

Greenville, North Carolina

Stephen F. Norton Editor

American Academy of Underwater Sciences 430 Nahant Road

Nahant, MA 01909-1696 www.AAUS.org

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Proceedings of the American Academy of Underwater Sciences 22nd Annual Scientific Diving Symposium “Diving for Science… 2003” Copyright © 2003 by American Academy of Underwater Sciences 430 Nahant Road Nahant, MA 01909-1696 www.AAUS.org No part of this book may be reproduced in any form without written permission from the Publishers. Copies of these Proceedings can be purchased from AAUS at the above address. Cover Photo: Doug Kesling. Divers: Tane’ Casserley and Steve Sellers. 2001 NOAA USS Monitor Project

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TABLE OF CONTENTS Page Introduction 1

Stephen F. Norton The Kill Zone 2

Rick Allen Reproductive Biology of the Gorgonian Leptogorgia hebes (Verrill) 3

S.E. Beasley, M.R. Dardeau, and W.W. Schroeder In situ Scuba Diver Identification of Hatchery Released Red Snapper, Lutjanus campechanus, Using Visual Implant Elastomer Tags in the Gulf of Mexico 19

Brett Blackburn, Nathan Brennan, and Ken Leber Anguilla’s Spanish Shipwrecks 20

Frank Cantelas Oyster Sanctuaries Construction with the Aid of SCUBA Diving 22

Clay Caroon A Ram Bow in the Keys: Latest findings from the Investigation of the Steamer Queen of Nassau 23

Tane Casserley Coastal Global Observing Systems: A Beginning 25

Robert Christian Wrecked, Abandoned, and Re-used: Archaeological Exploration on the Great Lakes 26

Annalies Corbin and Bradley A. Rodgers Untangling Damselfish Mortality from Movement Through the Use of Spatially-Explicit Data Collection and Analysis Techniques 27

Will F. Figueira and Sean J. Lyman ScapaMap: the Scapa Flow Maritime Archaeology Project 28

Bobby Forbes Predator Suites and Flabellinid Nudibranch Nematocyst Complements in the Gulf of Maine 37

Kinsey Frick Compelled to Run His Majesty's Ship Ashore: The Story of the HMS Santa Monica as Historical, Cultural and Environmental Resource 38

Kelly Gleason

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Scientific Diving Training through an Aquarium Diving Program 39

Roy Houston Submerged Aquatic Vegetation Assessment in Florida Estuaries: Methodology 40

Marc D. Julian, Gil McRae, Howie Brown, and Kevin Madley The Ph.D. Program in Coastal Resource Management at East Carolina University 42

Russ Lewis Considerations for Scientific Technical Diving: An Overview of Logistics, Procedures, and Implications for Program Development 43

Michael Lombardi Using Diving and Remote Sensing Approach to Classify Benthic Habitats in Coral Reef Ecosystems in Belize 59

Joseph J. Luczkovich and Jason W. Rueter Noisy Fish and even Louder Divers: Recording Fish Sounds Underwater, with some Problems and Solutions using Hydrophones, Sonobuoys, Divers, Underwater Video and ROVs 60

Joseph J. Luczkovich and Mark W. Sprague Hampster Balls, Voyeurism, and Video: A Method of Monitoring Reproductive Behavior in Coral Reef Fish 62

Sean J. Lyman and Will F. Figueira Scientific Diving Training through an Aquarium Diving Program 63

Peter Pehl Rubber Duckies or Shipwrecks: A Submerged Cultural Resource Survey of Bath, North Carolina 64

Andrew Pietruszka Transthoracic Echocardiography (TTE) - A Tool to Monitor Unsafe Decompression Stress 65

Neal W. Pollock Spatial and Seasonal Trends for Microbial Community Structure and Biomass across a Pollutant Gradient in the Western Basin of Lake Erie 72

J.A. Porter and R.H. Findlay

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PRISM Topaz Mixed-Gas Closed Circuit System: Design and Development For Recreational, Scientific, and Commercial Users 74

Peter F. Readey Shipwrecks and Students: Discovering Maritime Heritage 75

Timothy Runyan Safe Harbors for our Future: An Overview of Marine Protected Areas 76

Derek Smith and Kathy Ann Miller Diving on the Queen Anne’s Revenge 77

Chris Southerly and Jelep Gillman-Bryan Use of Scientific Diving in the Coastal Ocean Research Monitoring Program (CORMP): Onslow Bay, North Carolina 78

Jason Souza and David Wells Underwater Crime Scene Investigations (UCSI), a New Paradigm 79

Greg Santon New Insights into Sea Urchin Recruitment in the Gulf of Maine 80

Rebecca L Toppin and Larry G. Harris To Catch a Flounder: the Potential for Escapement of Older Southern Flounder (Paralichthys lethostigma) from Fishing Pressure in North Carolina 81

Carter Watterson and John Alexander What does a Sponge Eat? Examining Variability in Sponge Nutrition in the Florida Keys 82

Jeremy B. Weisz, Melissa Southwell, Christopher S. Martens, Niels Lindquist, and Ute Hentschel

The Introduction and Dispersal of the Indo-Pacific Lionfish (Pterois volitans) Along the Atlantic Coast of North America 84

Paula Whitfield, Todd Gardner, Stephen P. Vives, Matthew R. Gilligan, Walter R. Courtney Jr., G. Carleton Ray, and Jonathan A. Hare

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Introduction

Stephen F. Norton Environmental Studies Program

University of Puget Sound 1500 N. Warner St.

Tacoma, WA 98416 SNorton@ups.edu

These proceedings - “Diving for Science… 2003,” contain the abstracts and/or papers presented at the 22nd Annual Scientific Diving Symposium of the American Academy of Underwater Sciences. The meeting was hosted by East Carolina University at the Greenville Hilton, Greenville, NC March 14th and 15th 2003. The local organizing committee consisted of Steven Sellers, Frank Cantelas, Reide Corbett, Annalies Corbin, Neal Pollock, Thomas Skalko, Joe Luczkovich, and Steve Norton. Valuable assistance was provided by Gwen Bibbs, Eric Diaddorio, and Jennifer Dorton. We would like to thank the North Carolina Sea Grant College Program for their support in the printing of these proceedings, Kathy Johnston for providing original artwork for scholarship fund raising, and the other members of the Academy who provided direction and guidance .

This year’s annual symposium highlighted the diversity and continued vitality of the American Academy of Underwater Sciences. Presentations and posters demonstrated valuable scientific insights made possible by the application of SCUBA as a research tool. Others explored the integration of innovative remote sensing technologies with traditional underwater science techniques. Several presentations showcased the critical importance of scientific diving in understanding the historical record of maritime activities. Others discussed advances in SCUBA technology and understanding of diving physiology that hold the promise of extending the depth and duration limits of underwater exploration while improving safety. Finally, several presenters shared their experiences on organizational and training issues surrounding scientific diving programs.

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The Kill Zone

Rick Allen P.O. Box 53269

Fayetteville, NC 28304 nautilusvideo@earthlink.net

Abstract

False Bay, South Africa. This idyllic location perched at the tip of the world may be the most dangerous place on the planet. Here lurks the most feared predator known to man . . . the Great White shark. No where else on Earth can you regularly witness 16 foot Great White sharks catapulting into the air while stalking and killing their prey. Get a sneak peak at a new documentary for National Geographic as you meet the unrivaled acrobats of the shark world and the kings of the deadliest place on earth in “The Kill Zone.”

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Reproductive Biology of the Gorgonian Leptogorgia hebes (Verrill)

S. E. Beasley Department of Biological Sciences

University of South Alabama Mobile, Alabama 36688

M. R. Dardeau1*

Dauphin Island Sea Lab 101 Bienville Boulevard

Dauphin Island, Alabama 36528 *mdardeau@disl.org

W. W. Schroeder

Marine Science Program The University of Alabama and Dauphin Island Sea Lab

Dauphin Island, Alabama 36528

1Corresponding Author Abstract

A population of the gorgonian, Leptogorgia hebes, was sampled monthly between April 1991 and April 1992 from a hardbottom area approximately 28 km south-southeast of Dauphin Island, Alabama in the northern Gulf of Mexico. L. hebes, a gonochoric species with a sex ratio biased in favor of female colonies (1:2), had gonads concentrated in distal branches of colonies. Histological analysis revealed an annual cycle of gonadal development; oogenesis began in January while spermatogenesis lagged behind, with primary spermatocytes not appearing until April and May. Gonads developed through the warm summer months, reaching maxima in numbers and diameter just before spawning in late August or early September. Both eggs and sperm were released between August and September collections. Age was determined by counting axial rings which appeared to exhibit annual periodicity. Age at first reproduction for female colonies was two years while male colonies delayed reproduction until six years old. Introduction

Natural hard substrates throughout the northern Gulf of Mexico support numerous hard bottom communities (Rezak et al. 1985, Putt et al. 1986, Schroeder et al.1988, Gittings et al. 1992a, Thompson et al. 1999). Temperate hard bottom communities, including those in the northern Gulf of Mexico, could not sustain fisheries without substantial levels of net production or organic matter disbursement (Hopkinson et al. 1991). In these areas, the aborescent branching morphology of gorgonians provides

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much of the physical relief and biotic cover attractive to epibenthic marine invertebrates and demersal fishes. Populations of Leptogorgia hebes (Verrill) dominate shallow inner shelf hard bottom communities in the northeastern Gulf of Mexico (Mitchell et al. 1993) and are also known to occur off the eastern coast of the United States (Wendt et al. 1985). Mitchell et al. (1992, 1993) provided information on secondary production, age and growth of L. hebes; however, little else is known about its life history.

Reproductive strategies among modular, colonial organisms may vary widely (Coma et al. 1995a). This study describes the annual cycle of sexual reproduction in L. hebes including population sex ratio, age of reproductive viability, onset of gametogenesis, spawning period and location of reproductive activity within colonies. Knowledge of basic life history traits of dominant organisms in hard bottom communities can lead to a better understanding of how they maintain their populations. Specifically, reproduction is the first step in successful colonization of substrate, particularly in areas where habitat is covered and uncovered intermittently by storm events (Mitchell et al. 1993). Methods

The study site, in the southern Southeast Banks Area, is located approximately 28 km south-southeast of Dauphin Island, Alabama in the northern Gulf of Mexico (30 o 00’05”N, 87o57’03”W) (Figure 1). It occupies a region of approximately 1.0 km2, with a depth range of 25 to 27 m, and is comprised of small (up to 2 m), low-relief (<0.25 m) slabs of shelly sandstone and sandy mudstone scattered across a flat sandy bottom (Schroeder et al. 1988, Mitchell et al. 1993). Of the two species of gorgonians at the study site, Leptogorgia hebes is the most abundant, however, L. virgulata is also common (Mitchell et al. 1993).

Utilizing SCUBA, up to thirty colonies of Leptogorgia hebes were randomly chosen and collected monthly at Southeast Banks for a period of 13 months from April 1991 to April 1992 except for November 1991. While in the field, the corals were stored in sea water. To examine the reproductive cycle, colonies were placed in Helly’s fixative for histological analysis (Yevich and Barszcz 1977). Colonies were washed several times in fresh water over a 24 hr period, then stored in 70% undenatured ethanol.

After fixation, colony height was determined by measuring from the basal plate to the tip of the longest branch. An electronic caliper was used to measure the diameter of each colony halfway between the basal plate the first branch. A geomorphological method of assigned stream orders (Strahler 1952) was first used to quantify branching networks in gorgonians by Brazeau and Lasker (1988). Mitchell et al. (1993) applied the same geomorphological method to L. virgulata and L. hebes. The most distal branches are defined as first order branches (1o). When two 1o branches come together they form a second order branch (2o). Higher order branches occur only when two lower branches of the same order come together. This method was applied to examine differences in fecundity between branch orders.

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For histological analysis, 10 one-centimeter-long sections of each branch order

were randomly sampled from each colony. When less than 10 cm of branch of a given branch order was present, then the entire branch was sampled. Longitudinal sections were made of each one-centimeter branch sample. Branch sections were first stained using Cason’s trichome stain technique (Cason 1950). Infiltration of samples was carried out with Spurr’s resin. Embedded samples were polymerized for eight hours in a dry heat oven at 70o Celsius. Using a low speed saw, longitudinal thin sections were made of each embedded branch sample. These sections were then mounted on petrographic slides by applying ultra-violet (UV) cement. A light (brightfield) binocular microscope was used to manually count oocytes and spermaries present. The diameter of oocytes was determined using a calibrated ocular micrometer. Data Analysis

The mean number and mean diameter of oocytes and spermaries per polyp per colony from histological sections prepared from each colony were compared with the basal growth ring counts of the corresponding colony using linear regression analysis. The mean number of oocytes per polyp, as well as diameters of oocytes from each branch order of the corresponding colonies, was compared using Kruskal-Wallis one-way analysis of variance (ANOVA) nonparametric test on ranks because assumptions for parameteric tests were violated. Subsequently, Dunn’s all pairwise multiple comparisons test was used to define which groups differed from each other (p <0.05). The mean number of spermaries per polyp from each branch order of corresponding colonies was compared by performing a Kruskal-Wallis one-way ANOVA. This was followed by the application of the Student-Newman-Keuls method of all pairwise multiple comparisons to determine which groups differed significantly from each other (p < 0.05). Comparison of the mean diameter of spermaries from each branch order was made by using Kruskal-Wallis one-way ANOVA nonparametric test on ranks because assumptions for parametric tests were violated. A Chi-square test was performed to determine if there was a significant deviation from a 1:1 sex ratio. Results Gender and reproductive cycle

Colonies of Leptogorgia hebes from the northern Gulf of Mexico were gonochoric. Oocytes and spermaries were attached by a short mesogleal stalk to the mesenterial filaments of the gastric cavity. Gonads were present from April 1991 through September 1991 and January 1992 through April 1992. A total of 113 colonies were examined during these months. Of these colonies, 51.3% (n = 58) were female, 22.1% (n = 25) were male, and 26.6% (n = 30) were indeterminate. Onset and duration of gametogenesis was determined by the presence or absence of gonads in colonies selected at random from each month from April 1991 through April 1992 (Figure 2).

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Primary oocytes, having diameters between 15 µm and 30 µm were first observed

in colonies in January 1992, indicating the onset of oogenesis. When the 1991 and 1992 reproductive seasons were considered together, oocytes continued to increase in size through the spring and summer months, reaching maximum diameters in August 1991 (Figure 2). Although the number of oocytes per polyp ranged from zero to 18 in August 1991 (Figure 3), only one to three of these per polyp were mature. The mature oocytes had a mean diameter of 340 µm (n = 30 oocytes). In August 1991, two different size classes of oocytes were present but only the smaller ones remained in September. No oocytes were present in samples from October 1991 through December 1991. Two size classes of oocytes were again observed in January and February 1992. Smaller oocytes were round, with diameters of 15 to 30 µm, while oocytes greater than 30 µm were oval in shape with the nucleus located eccentrically.

Spermatogenesis lagged behind oogenesis, with primary spermatocytes not appearing until April and May (Figure 3). Spermaries were only present from April to September; maturing through the warm summer months (Figures 2, 3).

Fewer gonads were present in the September 1991 collection than in the August 1991 collection (Figure 3; p = 0.95, n = 29 for August colonies and n = 6 for September colonies). Gonads were significantly smaller in the September 1991 collection than in the August 1991 collection (Figure 2).

To establish sex ratios, all colonies (n = 30) from the peak reproductive period in August 1991 were used. The August 1991 collection contained 19 female colonies, 10 male colonies, and one colony whose sex could not be determined. A Chi-square test revealed that the sex ratio was significantly biased (p < 0.05; n = 29) in favor of female colonies 1:2. Distribution of gonads within colonies

During the month of peak gamete abundance per colony, August 1991, gamete production was concentrated in distal branches. Mean number of oocytes per polyp of 1o, 2o, and 3o branches differed significantly (p < 0.05) from 5o branches for female colonies (Figure 4). The mean number of oocytes per polyp did not differ between 4o and all other branch orders. For male colonies, 1o, 2o, and 3o branches had significantly more spermaries per polyp (p < 0.05) than 4o and 5o branches (Figure 5). Oocytes from the August 1991 collection were of similar sizes in all branch orders. Spermaries were also of similar sizes in all branch orders. Age and colony height at spawning

Ages of Leptogorgia hebes in the northern Gulf of Mexico for all colonies colleted from April 1991 through April 1992 ranged from 2 to 17 years. Female colonies with mature oocytes had a minimum of 2 and a maximum of 17 basal growth rings present suggesting that female colonies continue to be reproductively active with

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increasing age. Regression analysis revealed that the number of oocytes present in each polyp tended to increase as the number of basal growth rings increased (n = 47; p < 0.001).

Spermaries were not detected in colonies with fewer than six basal growth rings. Regression analysis failed to detect a relationship between increasing number of spermaries per polyp and an increase in number of basal growth rings for corresponding male colonies (n = 21; p = 0.109).

Heights of colonies from April 1991 through April 1992 ranged from 53 to 673 mm. From the August collection, oocytes and spermaries occurred in colonies having a minimum height of 81 mm and 150 mm respectively. Discussion

Leptogorgia hebes, like many other gorgonians (Kinzie 1971, Goldberg and Hamilton 1974, Grigg 1977, Brazeau 1986, Brazeau and Lasker 1990, Gotelli 1991, Coma et al. 1995a), is gonochoric, iteroparous and appears to have an annual spawning cycle. Primary oocytes, first discernable in January and early February, matured and reached maximum size in August. Spermatocytes first appeared in April and May and took only four to five months to reach maturity. This pattern of delayed spermatogenesis, so that spawning by both sexes coincides, is a common feature of gonadal development in all gorgonian species for which this information is available (Coma et al. 1995a).

Oocytes and spermaries generally increased in size in the four months prior to the single, synchronous spawning event suggested by the sudden reduction in numbers of gametes, followed closely by several months with no gametes present. Mean numbers of gonads per polyp, on the other hand, remained nearly constant in the months prior to spawning. Not all of the gonads were mature at the time of spawning and it seems unlikely that these 100 µm gonads matured and spawned between the August and September sampling. The smaller gonads still present in September may have degenerated and been resorbed by the polyps. About 30% of the oocytes were in a state of deterioration just prior to spawning in the gorgonians Muricea californica and M. fruticosa (Grigg 1979). It is unclear why certain oocytes develop to maturity and others do not, however, Campbell (1974) suggests that resorption of oocytes may provide nutrients for the rapid growth of the remaining oocytes.

Although gonad development was concurrent in all branch orders of colonies of L. hebes, reproductive effort was concentrated in polyps of the upper and central branches of both male and female colonies. Gonads occurred in greater numbers in polyps of 2o branches, but their numbers were significantly reduced in polyps of older branches found near the base of the colony. Beiring and Lasker (2000) also found a near absence of gametes in polyps from the bases of colonies of Plexaura flexuosa, and Coma et al. (1995b) noted a significant decrease in number of gonads with an increase in branch order in Paramuricea clavata. Fecundity may have been lower in these lower branches

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because, as the colony grows, the polyps changed their role in response to more efficient food capture by upper branches. Reduced reproductive effort of polyps at the growth tips in the gorgonian Briareum asbestinum has been reported as well (Brazeau and Lasker 1990). Colony growth occurs at the tips of 1o branches, thus diversion of energy from reproduction to growth may contribute to reduced fecundity of polyps in 1o branches.

Colony age and size also influenced reproductive effort. L. hebes delays

reproduction until a minimum age of two years in females and six in males, presumably allocating energy for growth rather than for reproduction during this period. This delay is relatively short compared to temperate gorgonians (5-13 years) and more similar to that of tropical species (2-5 years) according to data compiled by Coma et al. (1995a). Gotelli (1988), however, noted that oocytes were present in 2 year old L. virgulata, a congener which also occurs in the northern Gulf of Mexico. L. hebes fit the general pattern among gorgonians of increased fecundity with increased size. Not only is there an increase in gamete production per polyp with an increase in colony size but, as polyps are added exponentially with growth, each is potentially capable of reproduction. Coma et al. (1995b) and Beiring and Lasker (2000) found that the larger individuals representing small percentages of populations of Paramuricea clavata and Plexaura flexuosa produced between 33-98% of the gametes, pointing up the importance of colony size structure in determining reproductive potential.

The six year delay in maturation of males and the sex ratio biased 1:2 in favor of females are characteristics not found in other gorgonian populations. With the exception of Briareum asbestinum, which has a population which favors males 2.2:1, all populations of gorgonians studied to date have 1:1 sex ratios (Coma et al. 1995a). No other studies have noted a significant delay in sexual maturity of male gorgonians. The female biased sex ratio and late maturing male population suggest that sperm limitation, the potential for unfertilized eggs due to dilution, is not a factor.

This study was not able to determine mode of reproduction or fertilization. Two modes of sexual reproduction occur among gonochoric corals: internal fertilization (only spermatozoa are released) followed by release of embryos after brooding of the planula larvae within the polyp, or broadcast spawning (both ova and spermatozoa are released) followed by external fertilization. Brooding corals have an extended breeding season (Harrison and Wallace 1990) and facilitate localized recruitment. Corals exhibiting broadcast spawning have a distinct spawning period (Brazeau and Lasker 1989, Harrison and Wallace 1990) and external development that encourages dispersal of gametes (Stimson 1978). Coma et al. (1995a) generalizes that tropical gorgonians tend to either broadcast spawning and external fertilization or asexual reproduction while temperate species tend toward internal fertilization and brooding.

Synchronous spawning, or compression of spawning into a short period around the same time each year, to maximize fertilization implies an environmental stimulus. Lunar patterns (Brazeau and Lasker 1989, 1990, Coma et al. 1995a) and sea water temperatures (Grigg 1977) are exogenous clues proposed to determine timing of spawning events in gorgonians and both could have played a role in initiating spawning

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(Hagman et al. 1998). Spawning by L. hebes occurred in late August or early September, the period of maximum seawater temperatures in the northern Gulf of Mexico (Thompson and Leming 1978). The August collection took place 10 days prior to the August full moon. The September collection was made 18 days after the August full moon and two weeks before the September full moon. The timing of this spawning event coincides with coral mass spawning events during the weeks following full moons in August and September in the northwestern Gulf of Mexico (Gittings et al. 1992b), Curaçao (Van Veghel 1993) and the Caribbean (Szmant 1991). Although spawning of L. hebes was not directly observed, the sudden disappearance of mature gametes and the timing of the spawn argue for broadcast spawning in this species.

Despite the obvious potential advantage of vegetative propagation in an environment where habitat is distributed in a very patchy manner, no evidence of either fragmentation or stolonization was observed. The early maturation of the colonies and the large number of gametes per polyp produced annually represent a heavy investment in sexual products that suggests sexual reproduction is important in maintaining the population. Acknowledgements

This research was supported in part by the NOAA Office of Sea Grant, Department of Commerce, under grant NA89AA-D-SG016 for project R/ER-19, the Mississippi-Alabama Sea Grant Consortium, the Dauphin Island Sea Lab, and the University of Alabama. We would like to thank John Dindo and Al Gunter for the long hours spent helping collect the gorgonians, Frederick Bayer for confirming the identification of Leptogorgia hebes, and the DISL vessel operations personnel. This publication is contribution 345 from the Dauphin Island Sea Lab, Dauphin Island, Alabama. References Beiring, E. and H. R. Lasker. 2000. Egg production by colonies of a gorgonian coral.

Mar. Ecol. Prog. Ser. 196:169-177. Brazeau, D. A. 1986. Male biased sex ratio in the Caribbean octocoral Briareum

asbestinum. Am. Zool. 26:7A. Brazeau, D. A. and H. R. Lasker. 1988. Inter- and intraspecific variation in gorgonian

colony morphology: quantifying branching patterns in arborescent animals. Coral Reefs 7:139-143.

Brazeau, D. A. and H. R. Lasker. 1989. The reproductive cycle and spawning in a

Caribbean gorgonian. Biol. Bull. 176:1-7.

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Brazeau, D. A. and H. R. Lasker. 1990. Sexual reproduction and external brooding by the Caribbean gorgonian Briareum asbestinum. Mar. Biol. 104:465-474.

Campbell, R. D. 1974. Cnidaria. Pp. 133-200, In: A. C. Giese and J. S. Pearse (Eds.),

Reproduction of marine invertebrates, Vol. I. Academic Press, New York. Cason, J. E. 1950. A rapid one-step Mallory-Heidenhain stain for connective tissue.

Stain Tech. 24:225-226. Coma, R., M. Ribes, M. Zabala, J-M. Gili. 1995a. Reproduction and cycle of gonal

development in the Mediterranean gorgonian Paramuricea clavata. Mar. Ecol. Prog. Ser. 117:173-183.

Coma, R., M. Zabala, J-M. Gili. 1995b. Sexual reproductive effort in the Mediterranean

gorgonian Paramuricea clavata. Mar. Ecol. Prog. Ser. 117:185-192. Gittings, S. R., T. J. Bright, W. W. Schroeder, W. W. Sager, J. S. Laswell and R. Rezak.

1992a. Invertebrate assemblages and ecological controls on topographic features in the northeast Gulf of Mexico. Bull. Mar. Sci. 50:435-455.

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Bright. 1992b. Mass spawning and reproductive viability of reef corals at the East Flower Garden Bank, northwest Gulf of Mexico. Bull. Mar. Sci. 51:420-428.

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University of Miami Rosenstiel School of Marine and Atmospheric Sciences, Miami, Florida. No. 1713, Pp 58-61.

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distribution of a shallow-water gorgonian. Ecol. 69:157-166. Gotelli, N. J. 1991. Demographic models for Leptogorgia virgulata, a shallow-water

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to vertical and geographical distribution. Pp. 41-59, In: S. E. Stancyk (Ed.), Reproductive ecology of marine invertebrates. University of S. Carolina Press, Columbia.

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scleractinian corals. Pp. 133-2707, In: Z. Dubinsky (Ed.), Ecosystems of the world, Vol. 25. Coral Reefs. Elsevier, Amsterdam.

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Hagman, D. K., S. R. Gittings and K. J. P. Deslarzes. 1998. Timing, species participation and environmental factors influencing mass spawning at the Flower Garden Banks (northwest Gulf of Mexico). Gulf of Mexico Science 16:170-179.

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Discovery Bay, Jamaica. Dissertation. Yale University, USA. Mitchell, N. D., M. R. Dardeau, W. W. Schroeder and A. C. Benke. 1992. Secondary

production of gorgonian corals in the northern Gulf of Mexico. Mar. Ecol. Prog. Ser. 87:275-281.

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structure, and relative growth of two gorgonian corals, Leptogorgia hebes (Verrill) and Leptogorgia virgulata (Lamarck), from the northern Gulf of Mexico. Coral Reefs 12:65-70.

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biota associated with natural hardbottom areas in the northwestern Gulf of Mexico. NE Gulf Science 8:51-63.

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Gulf of Mexico. John Wiley & Sons, New York. Schroeder, W. W. A. W. Shultz and J. J. Dindo. 1988. Inner-shelf hardbottom areas,

northeastern Gulf of Mexico. Trans. Gulf Coast Assoc. Geol. Soc. 38:535-541. Stimson, J. S. 1978. Mode and timing of reproduction in some common hermatypic

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annularis and M. cavernosa. Mar. Ecol. Prog. Ser. 74:13-25. Thompson, P. A. and T. D. Leming. 1987. Seasonal description of winds and surface

and bottom salinities and temperatures in the northern Gulf of Mexico, October 1972 to January 1976. NOAA Tech. Rep. NMFS SSRF-719. 44 p.

Thompson, M. J., W. W. Schroeder and N. W. Phillips. 1999. Ecology of live bottom

habitats of the northeastern Gulf of Mexico: A community profile. U. S. Dept. of the Interior, U. S. Geological Survey, Biological Resources Division, USGS/BRD/CR—

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1999-001 and Minerals Management Service, Gulf of Mexico OCS Region, New Orleans, LA, OCT Study MMS 99-0004. x pp. + 74 pp.

Van Veghel, M. L. J. 1993. Multiple species spawning on Curaçao reefs. Bull. Mar.

Sci. 52:1017-1021. Wendt, P. H., R. F. van Dolah and C. B. O’Rourke. 1985. A comparative study of the

invertebrate macrofauna associated with seven sponge and coral species collected from the South Atlantic Bight. Jour. Elisha Mitch. Sci. Soc. 101:187-203.

Yevich, P. and C. A. Barszcz. 1977. Preparation of aquatic animals for histopathological

examination. Manual E. P. A. Environmental Research Laboratory, Narragansett.

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Figure 1. Location of Southeast Banks Area.

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Figure 2. Mean diameter of gonads from April 1991 through April 1992 (n=number of

gonads measured, bars represent standard error of the mean. Oocytes were not present October – December. Spermaries were not present October – March.

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Figure 3. Mean number of gonads per polyp from April 1991 through April 1992

(n=number of colonies of each sex observed with gonads, bars represent standard error of the mean). Oocytes were not present October – December. Spermaries were not present October – March.

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Figure 4. Mean number of oocytes per polyp with respect to branch order (n=number of

polyps observed, bars indicate standard error of the mean).

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Figure 5. Mean number of spermaries per polyp with respect to branch order (n=number

of polyps observed, bars indicate standard error of the mean).

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In situ Scuba Diver Identification of Hatchery Released Red Snapper, Lutjanus campechanus, Using Visual Implant Elastomer Tags in the Gulf of Mexico

Brett Ramey Blackburn* Nathan Brennan

Ken Leber Mote Marine Laboratory

1600 Ken Thompson Parkway Sarasota, FL 34236 *Bdiver@mote.org

Abstract

Over the last 6 years approximately 3000 juvenile red snapper have been release on artificial reef habitats off the coast of Sarasota, FL. The releases were conducted to investigate the use of hatchery reared juvenile red snapper to supplement native populations in the Gulf of Mexico. Pilot releases were conducted in 2000 and 2002 to investigate release habitat preferences, stocking density effects, and in situ acclimation techniques to develop stocking and release protocols for red snapper. Post release assessments were based around identification and enumeration of released fish. Visual implant elastomer tags of different colors from Northwest Marine Technology, Lopez Is., WA were used to identify experimental treatments of released fish. Elastomer tags implanted between the caudal fin and anal fin rays provide a good short-term (6-8 months) identification mark for diver identifications. This tagging technique was only restricted by the number of highly contrasting colors available for comparison; however fluorescent colors were easiest to distinguish. Identification of tag lots under low light conditions was restrictive when comparing black, blue and purple. Yellow and green combinations were also difficult to distinguish under low natural lighting. The use of a low power dive light helped to differentiate the colors in these situations. Complicated experimental designs with multiple release sites and treatments required the use of the anal fin in conjunction with caudal tagging (16 tag codes). In these experiments, tags in the caudal fins were used to identify the release sites while anal tags implanted in the fins identified individual treatment types. The high number of tag codes could result in decreased abilities of the divers to identify correct codes; however, visual implant elastomer tags provide an excellent monitoring technique for in situ monitoring of released fish by divers.

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Anguilla's Spanish Shipwrecks

Frank Cantelas Maritime Studies Program

East Carolina University Greenville, NC 27858

Cantelasf@mail.edu.edu Abstract

In the early hours of July 8, 1772 the sounds of timber smashing, rigging falling and the shouts of men echoed on the desolate shore near the east end of Anguilla, a small island in the British West Indies. Two ships, the 70 gun man-of-war El Buen Consejo, and the 40 gun armed merchant vessel Prusiano, part of a larger Spanish supply fleet headed for Vera Cruz, made a terrible navigation error as they neared the end of a long trans-Atlantic voyage. While making for Anagada Passage to enter the Caribbean, they missed a course change that led to an abrupt collision with the island. Anchors were let go when a lookout heard the crashing surf but by then it was too late - they grounded on the rocky shore.

Fortunately for the crew and passengers, calm seas allowed everyone to escape without loss. But suddenly the small British island was overwhelmed by the unexpected influx of nearly one thousand Spaniards. El Buen Consejo also carried a contingent of Franciscan monks on their way to the Pacific. Among their belongings were many brass religious medallions for use in missionary activities. Anguilla is a small arid island and the additional people strained the inhabitant's meager resources. British troops were called in to keep the peace until the shipwreck survivors were moved off the island in November and taken to Puerto Rico.

The ships, however, did not survive. They settled after grounding in shallow water, allowing the Spaniards to salvage much of the cargo and armament. These supplies probably eased the living conditions of the survivors. Inevitably, as the hurricane season progressed, a storm approached the island and the vessels were pushed back into deeper water where they broke apart and sank.

The story of the shipwrecks is well known in Anguilla but the ships themselves were only discovered in recent years. In 1996 the Anguillian Government invited the Maritime Studies Program at East Carolina University (ECU) to examine the remains of the two vessels and offer suggestions for their future management as cultural resources. ECU completed archaeological documentation of El Buen Consejo and a reconnaissance of the Prusiano. This study discovered evidence of the unfolding drama of the crew trying to save the ships as they neared the island. The anchors failed to set on the rocky bottom and clusters of iron fasteners concreted on the rocky shore point to where one of the vessels struck. Other remains illustrate the destructive force of the hurricane that

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broke the ships apart. A trail of eighteen cannon and mounds of concreted cargo packages show how the bottom was torn out of El Buena Consejo as it was pulled off the shore.

The legacy of the Spanish shipwrecks is found in the island's waters and is an important component of Anguilla's heritage. The well-known story and its archaeological record is the subject of this presentation.

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Oyster Sanctuaries Construction with the Aid of SCUBA Diving

Clay Caroon North Carolina Division of Marine Fisheries

3441 Arendell St. P.O. Box 769

Morehead City, NC 28557 Clay.Caroon@ncmail.net

Abstract

In the mid-1990’s, the Artificial Reef Program and the Shellfish Rehabilitation Program cooperatively constructed five Oyster Sanctuaries/Artificial Reefs. The five sites are constructed of Class B limestone rip-rap (football-basketball sized rock) and are located in North Carolina’s estuaries. Initial colonization showed promise, but later recruitment success was below expectations. Initially recruitment surveys were conducted by scuba divers sending rip-rap material to the surface for a visual inspection. Species counts were made and the material was placed back on the sanctuaries.

Several years lapsed without any inspections of the sanctuaries. In 2002, relief money was available from the National Marine Fisheries Service Grant for Hurricane Floyd damages. Three of the existing oyster sanctuaries were to be enhanced by over-planting with three different cultch materials. However, preliminary site surveys conduced by scuba divers found oyster densities high enough to prohibit over-planting. Instead of over-planting existing material, a new “leg” will be constructed on each of the three sits. The new legs will be constructed by creating mounds with class B rip-rap. While each of the mounds is being constructed, the shape, size, and height will be monitored by scuba divers. This will create a base for the cultch material over-planting. The construction of a new leg also provides the ability to leave a bare control area of fresh Class B rip-rap to be compared with the three over-planted treatments. The adjacent bottom will also receive treatments of the three cultch materials. This will provide information on the advantages of high profile construction techniques and whether the application of a veneer of cultch material will increase recruitment. The project will also help determine differences in effectiveness of various cultch materials and if it varies regionally. Each of the above plantings, as well as the control areas, will be monitored at regular intervals by scuba divers. Biological data will be recorded and comparison will be made.

This project will provide protected habitat for oysters, allowing brood stock development while providing information on methods for future sanctuaries. The next phase of the grant will provide the material to construct three new sanctuaries – utilizing information gained from the study. The last phase will be to construct three sites for harvest in the area of the sanctuaries, which could gain the benefit of sanctuary brood stock spawn.

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A Ram Bow in the Keys: Latest findings from the Investigation of the Steamer Queen of Nassau

Tane Casserley Monitor National Marine Sanctuary

100 Museum Drive Newport News, VA. 23606

Tane.Casserley@NOAA.gov

Abstract

This paper presents a comprehensive examination of the steamer Queen of Nassau, former (Canadian Government Ship) CGS Canada. It will demonstrate that this twentieth century vessel was one of the most influential ships in Canadian history. Although the British Royal Navy guarded Canada’s coastal waters from colonial times, beginning in the mid-nineteenth century the two governments argued over who should ultimately be responsible for Canada’s naval defenses. A growing naval threat in Europe at the dawn of the twentieth century placed increasing stress on the Royal Navy, while at the same time, competition for the economic resources of the North Atlantic placed pressure on Canada’s small and aging Fisheries Protection fleet. The construction of the CGS Canada was a direct result of these dynamics. This paper will establish the importance of this multi-function vessel, which embodied the new nation’s need for fisheries protection, coastal defense, and police work.

The steamer CGS Canada was built in 1904 and became the first armed, steel-hulled cruiser owned and operated by the Canadian government. The Canada’s ram bow, 10-to-1 length-to-beam ratio, and steel hull were a departure from the previous style of Canadian armed vessels. Consequently, the Canada marked the transition from traditional wooden schooners to modern steel cruisers, playing a crucial role as Canada formulated its young navy. The Canada was the fastest ship in the Fisheries Protection fleet; it was Canada’s first successful naval training vessel, and the first Canadian naval vessel to train with the Royal Navy.

In 1924, the Canada was sold to Barron Collier, a wealthy Florida landowner. Collier renamed the vessel Queen of Nassau and used it as an inter-island cruise ship for the lucrative Nassau-Miami route. Failing financially in its new role, the vessel sank under mysterious circumstances on July 2, 1926.

Recreational divers discovered the wreck in 2001, approximately seven miles south of Lower Matecumbe Key within the Florida Keys National Marine Sanctuary, and reported their find to Sanctuary officials. The site is now the focus of an ongoing archaeological investigation by a NOAA team consisting of the Monitor National Marine Sanctuary, East Carolina University, and the National Undersea Research Center at the

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University of North Carolina-Wilmington. The vessel is in remarkably good condition, lying intact on top of the sand in 230 feet of water.

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Coastal Global Observing Systems: A Beginning

Robert R. Christian Biology Department

East Carolina University Greenville, NC 27858

ChristianR@mail.ecu.edu Abstract

The United Nations has begun addressing the detection and prediction of global and large-scale change through the development of observing systems. These observing systems build on the activities of government and non-government programs that make long-term measurements. The function of the observing systems is to link the data from these programs to organizations that could use the integrated data and data products. Products include models that help to predict what and why changes occur beyond single nation boundaries. The three observing systems are the Global Climate Observing System (GCOS); the Global Oceanographic Observing System (GOOS); and the Global Terrestrial Observing System (GTOS). The interface between the land and ocean provides complexities that affect the capabilities of the three global observing systems. Understanding the three-way and complex interactions between natural and human systems at various scales is central to successful management of the coast in the face of change. Thus, there is a clear and recognized need to address the coastal zone in an integrated fashion within the observing system framework. Both GOOS and GTOS have begun to organize initiatives for the coast, with the GOOS initiative being more developed. They will use the observing system philosophy to identify and improve access to data and information about coastal change; enhance the capacity of the developing world to collect and manage information; assist users to make that access systematically; ensure that appropriate measures are being or can be made; and integrate terrestrial observations with marine observations.

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Wrecked, Abandoned, and Re-used: Archaeological Exploration on the Great Lakes

Annalies Corbin* Bradley A. Rodgers

Maritime Studies Program East Carolina University

Greenville, NC 27858 *Corbina@mail.ecu.edu

Abstract

The Great Lakes of North America comprise nearly 100,000 square miles (160,934 km) of fresh water and stretch inland for nearly 1,000 miles (1,609 km). This is a fair sized sea by any definition. The unique properties of these inland seas have shaped their maritime history and helped builders on the lakes produce both unique craft as well as perfect and adapt ocean going technologies. For more than a decade, the Maritime Studies Program at East Carolina University (E.C.U.) has studied the maritime history of this inland sea by examining the shipwrecks, abandoned, and re-used vessels now scattered across the lake floors. Of primary interest to researchers at E.C.U. has been the everyday work horse or vernacular watercraft of the Great Lakes. From early coasting haulers to the giant bulk carriers it is through a careful examination of these “less than romantic” relics that we are able to understand and interpret the maritime culture of the Great Lakes.

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Untangling Damselfish Mortality from Movement Through the Use of Spatially-Explicit Data Collection and Analysis Techniques

Will F. Figueira Duke Marine Laboratory

135 Duke Marine Lab Rd. Beaufort, NC 28516

Wff@duke.edu

Sean J. Lyman Duke University Hyperbaric Service

0584 Clin Res II DS Durham, NC 27710

Abstract

Obtaining accurate estimates of mortality for relatively sedentary organisms can often be confounded by the movement of individuals into and out of the study area. Here we present the results of a mortality study on the bicolor damselfish, Stegastes partitus, in which these problems were addressed through the use of spatial data collection and analysis techniques. For 2 years we followed several cohorts of uniquely tagged fish located in 2 different habitat types of the Florida Keys reef tract. Underwater mapping techniques were used to track capture and recapture locations to within approximately 1 meter. Using several spatial analysis techniques we were able to estimate the degree of movement occurring in the different habitats and used this information to correct the mortality estimates using Jackson’s square within a square technique. This study highlights the use of spatially-explicit data collection techniques in underwater research and demonstrates their use in post-hoc corrections of mortality estimates to account for movement that may have been greater than expected.

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ScapaMap: the Scapa Flow Maritime Archaeology Project

Bobby Forbes Dive Unit

Heriot-Watt University Old Academy Back Road

Stromness, ORKNEY. U.K. KW16 3AW B.Forbes@hw.ac.uk

Abstract

The naval wrecks of the Scapa Flow form a unique underwater record of one of the great periods of British and German maritime history. The ships of the German High Seas Fleet, in particular, fought through the Great War, ending in Scapa Flow in internment. Under the misapprehension that the armistice talks were about to fail and the 74 vessels would be used against Germany in future conflicts, German officers and crew scuttled much of the fleet on 21st June 1919. Seven wrecks of major warships remain, up to 25,000 tons and over 200m in length, in an area of 8 km2 of relatively flat, muddy seabed, in 30-50m depth. In between them lie concentrations of other wreckage associated with salvage activities on the existing wrecks and the vessels, which were raised and subsequently scrapped.

The remains of the High Seas Fleet represent an archaeological and historical

resource of hitherto unrealised potential, having been the subject of a wide range of interests in the past from salvage to recreational diving.

In the early 90s, Historic Scotland was approached by a variety of people and

organisations, worried about souvenir hunting and the reported deterioration of the Scapa Flow wrecks:

• Scottish Museums Council, for instance. Lyness Museum was being inundated

with large trophies, too big for the divers to take home, dumped on the pier and euphemistically called 'donations';

• Orkney bodies, including the local Council, aware that the wrecks were becoming a diminishing asset;

• ex-naval and military bodies, wanting to see the remnants of the military history of the Flow conserved.

The sites themselves lie in an environment, which is itself constantly changing

and subject to a variety of natural and man-made impacts. Effective maritime archaeological management requires base maps of the archaeological resource and quantified data on associated marine environmental parameters such as habitats, corrosion potential, sediment characteristics and behaviour, water quality, and factors

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such as impacts from visitors and nearby development. Although the archaeological potential and sensitivity of the remains of the High Seas Fleet is regarded as being significant, basic maps of the area and the individual sites were not available. Secondly, the baseline data on which to build effective monitoring strategies of the condition of these sites in the future is similarly unavailable. Thirdly, no periodic monitoring programme has been put forward which would provide quantitative data on processes affecting the sites.

Consequently, the ScapaMAP project was launched with the aims of:

• collating all possible archive materials; • assess the current condition of the sites; • promote future protection; • encourage access by the widest possible community

In addition, it had been decided to schedule the wrecks under the Ancient

Monuments and Archaeological Areas Act 1979 and timed with the closing phase of the project. On the 23 May 2001 four cruisers and three battleships were entered into the Schedule of Monuments. The Act still allows access to the sites but it is an offence to cause any damage to a Scheduled Monument.

Due to the area over which the remaining wreck sites are located, the size of the

wrecks themselves, the depths involved, time and budget constrains a variety of techniques were employed to fulfil the original objectives of the project.

Initial research of the data archive revealed an abundance of pictorial material centred around the scuttling and initial salvage operations and several historic account of these events (George 1973, Bowman 1964, van der Vat 1982). Several dive guides have also been written about the remaining wrecks. However, reports suggested that in recent years the wrecks had undergone considerable deterioration and the information contained in these reports was inaccurate. (Ferguson, 1985, Ferguson 1988, Smith 1989, MacDonald 1990)

Remote sensing techniques were used to develop base maps of the sites and the area as a whole and of each wreck site. In 1999 an impromptu survey was carried out using a Klein 2000 side scan sonar. The following year a magnetometer and a side scan sonar survey was conducted using an IMAGENEX 858 system operating at 675 kHz. A number of images were also donated to the project by Halliburton SubSea Ltd, following a training course using the Simrad EM3000 shallow-water multibeam system. Preliminary site investigations were carried out by divers from the University Dive Unit and the Archaeological Dive Unit, which works in collaboration with Government bodies to help preserve our underwater heritage. Both SCUBA and Surface supplied techniques using Enriched Air Nitrox were employed. Divers obtained both stills images and video for subsequent mosaicing.

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In 2001, a high resolution swathe bathymetry survey was carried out using the Reson SeaBat 8125. This is a wide-sector, wide-band focused multibeam sonar utilizing 240 dynamically focused receive beams. The system measures a 120º swath across the seafloor (3.5 times the depth of the water column), detects the bottom, and delivers the measured ranges at a depth resolution of 6 mm. The backscatter intensity image is displayed in real time on the sonar display. The SeaBat 8125 can be controlled through its native graphical user interface, or from a multibeam data collection system, such as the SeaBat 6042 version 7.

The Data Capture Station comprised of a Reson 81P topside processor, 6042 data

capture system and TSS POS/MV320 positioning system. Survey planning and processing station comprised of laptops used for planning survey lines, feeding them to the helm for ship control and to post-process the data from the previous lane. Data was subsequently corrected for tidal variation. Corrected data could then be viewed using GeoZui3D (Georeferenced Zooming user interface 3D) developed at the Center for Coastal and Ocean Mapping, University of New Hampshire.

The remote sensing surveys provided a variety of base maps of varying degrees of

accuracy, the highest resolution obtained using the Reson SeaBat system and the associated positioning system. All provided a valuable tool in the planning diving operations optimising dive time. In addition, the data acquired by the SeaBat system, when view with 3D visualisation software, provides a valuable education tool for both divers and non-divers. Current work includes incorporation of this data into an updated divers guide and for interpretation displays at local museums to promote a wider appreciation for the conservation of our underwater heritage.

The final report on the ScapaMAP project was submitted to Historic Scotland

detailing possible management and monitoring strategise for the Scheduled Monuments. A development of the project is the training of recreational divers through the Nautical Archaeological Society’s “Dive With a Purpose Programme”. Data gathered from during this work will be incorporated into the Royal Commission for Archaeological and Historic Monuments of Scotland (RCAHMS) CANMORE and CANMAP data archive.

The author is particularly grateful to Brian Calder (University of New Hampshire), Richard Lear (Reson UK Ltd) and members of the Archaeological Dive Unit (St Andrews University). Funding for the ScapaMAP project was provided by Historic Scotland and the Carnegie Trust for the Universities. Literature Cited Bowman, G. 1998. The Man Who Bought a Navy. G.G. Harrap & Co Ltd. Ferguson, D.M. 1985. The Wrecks of Scapa Flow. The Orcadian, Kirkwall. Ferguson, D.M. 1988. Shipwrecks of Orkney, Shetland and Pentland Firth. David and

Charles, Devon.

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George, S. C. 1999. Jutland to Junkyard. Birlinn Ltd., Edinburgh. MacDonald, R. 1990. Dive Scapa Flow. Mainstream Publishing, Edinburgh. Smith, P.L. 1989. The Naval Wrecks of Scapa Flow. The Orkney Press. ISBN: Van Der Vat, D. 1988. The Grand Scuttle. The Sinking of the German Fleet at Scapa

Flow in 1919. Grafton Books.

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Figure 1: Interned German High Seas Fleet at anchor in Scapa Flow, Orkney. Photograph taken from Houton with the island of Cava in the centre of the image. Image courtesy of Kirkwall Library.

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Figure 2. IMAGENEX 858 screen image of the SMS Koln. Bow is at the top left of the image with the vessel lying on her starboard side.

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Figure 3. Simple hand crafted mosaic of the plate separation of the plates on the port side near the bow of the SMS Brummer. The image is composed of two stills images corrected for optical distortion at the edges of each frame

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Fbth

igure 4. Mosaiced image constructed from miniDV video shot of the area between the ridge and the bow on the SMS Koln. Image shows the deterioration of the plates along e port side where it joints to the deck.

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Figure 5. Captured image of the SMS Koln obtained using GeoZui3D visualiser looking from bow to stern.

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Predator Suites and Flabellinid Nudibranch Nematocyst Complements in the Gulf of Maine

Kinsey Frick Department of Zoology

University of New Hampshire 46 College Rd.

Durham, NH 03824 Kinseym@cisunix.unh.edu

Abstract

Nematocysts are a primary defense mechanism for combating predation in aeolid nudibranchs, and predator presence influences nematocyst uptake through nudibranch response to chemical cues from predators. However, the degree of predation pressure on individual nudibranchs within a population is not uniform across a species range, particularly for those that are widely distributed. Nematocyst selection in a given nudibranch species may differ latitudinally or based on predator suites at different collection locations. Given the geographic ranges of predatory species in the area and extreme changes in water temperature, nudibranch species are potentially exposed to different predator suites between northern and southern regions even within the relatively small Gulf of Maine.

I used subtidal band transects to assess predator suites in different regions within the Gulf of Maine and collected aeolid nudibranchs of the genus Flabellina from these sites to analyze nematocyst types conserved by the nudibranchs. Nudibranchs analyzed include both generalist and specialist predators with varying degrees of ability to respond to predator cues due to variation in the nematocysts available in their cnidarian prey sources. Predator suites between northern and southern regions were statistically independent, with differences in number, density, and presence of specific predators. Some collected nudibranch species showed correlative differences in their nematocyst complements. The degree of response depended on the feeding specificity of the nudibranch and associated limitations in available nematocysts. Nematocyst selection may relate to an inducible defense in response to the presence of predators.

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Compelled to Run His Majesty's Ship Ashore: The Story of the HMS Santa Monica as Historical, Cultural and Environmental Resource

Kelly Gleason Maritime Studies Program

East Carolina University 1794 East 3rd St.

Greenville, NC 27858 Kag1122@mail.ecu.edu

Abstract

In April of 1782, Captain John Linzee sailed the 36-gun frigate HMS Santa Monica towards Tortola in the British Virgin Islands following raids on the island by American Colonists. Captain Linzee was sailing the HMS Santa Monica, a prize of war captured by the HMS Pearl during the siege of Gibraltar in 1779. On April 1st, the 145 foot British frigate was deliberately run aground on the island of St. John to keep her from sinking. Captain Linzee salvaged what he could and the wreck was, for the most part, forgotten for about 188 years. The HMS Santa Monica is a reminder of the often-disregarded role that the Caribbean played during the American Revolution. Management of a submerged cultural resource is a delicate balance of cooperation, outreach and protection. Allowing the interested public access to the remains of the HMS Santa Monica and still treating the site with integrity as an ecosystem is a dilemma for managers. The story of the HMS Santa Monica should be told, but not without careful attention to the impacts the “hands-on” history of archaeological research will have. Research conducted on the site in June of 2002 attempted to determine the impact that archaeological research has upon a coral reef ecosystem, such as the one that the HMS Santa Monica has become a part of while it still remains largely intact on the ocean floor after over 200 years. The site is an excellent example of interdisciplinary management, as the wreck of the HMS Santa Monica is not only a valuable part of maritime history, but also an important cultural and natural resource to be managed with care.

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Scientific Diving Training through an Aquarium Diving Program

Roy Houston Seaver Science Hall

Loyola Marymount University One LMU Drive

MS 8220 Los Angeles, CA 90045-2659

Rhouston@lmu.edu Abstract

The Aquarium of the Pacific in Long Beach, California maintains an active and experienced diver program that includes both staff and volunteers. Within the AoP Dive program is the Scientific Diver Course, which is an introduction to basic underwater sampling protocols that are widely used in environmental monitoring. In addition, the student gains a working knowledge of scientific methodology including experimental design, data collection, analysis and interpretation. A research paper is a fundamental requirement for completion of the course. With this training these divers are an asset, which is readily utilized by university research programs. Peter Pehl II, Dive Safety Officer of the Aquarium of the Pacific and Roy S. Houston, Ph.D. of Loyola Marymount University, will give consecutive presentations on the scientific diving program.

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Submerged Aquatic Vegetation Assessment in Florida Estuaries: Methodology

Marc D. Julian* Gil McRae

Howie Brown Kevin Madley

Florida Marine Research Institute Florida Fish and Wildlife Conservation Commission

100 8th Avenue SE St. Petersburg, FL 33701-5095 USA

*marc.julian@fwc.state.fl.us Program Design

Florida’s Inshore-Marine Monitoring and Assessment Program (IMAP) is a five-year project, which samples 180 statewide sites annually. Florida's inshore marine resources are one of the state's most valuable assets. They are unique and diverse, ranging from major embayments and lagoons to smaller estuaries at river mouths to tidal marshes and mangrove forests, which merge directly with the sea. IMAP is a collaborative project between EPA and the FWC Florida Marine Research Institute (FMRI) designed to assess the ecological condition of Florida’s inshore waters using a set of ecological indicators. IMAP samples on two different scales: statewide and regional. The regions correspond to Florida’s five Water Management Districts (WMD’s). At both scales, sample sites are determined randomly within a network of hexagonal grids. IMAP does not have the resources to characterize the variation in each indicator over fine spatial and temporal scales. Because of this limitation, the IMAP sampling design focuses on characterizing spatial differences. IMAP samples annually, and covers several locations throughout the state during each sampling period. In order for this strategy to work, there must be an identifiable time window (an index period) within which inshore resources are under significant stress, and conditions are not changing dramatically. Although there are exceptions, for most of Florida, a late summer index period meets these criteria. Under this framework, IMAP samples 180 stations each year during a concentrated late-summer index period. IMAP Indicators

IMAP indicators are environmental measurements that are used to reflect the quality of the ecosystem from which they are sampled. It is critical that these indicators are 1) responsive to environmental problems (such as pollution or nutrient enrichment), 2) easily measured, and 3) representative of the area being sampled. Highly mobile organisms or those that are tolerant of environmental pollution are not good choices in this type of program. The indicators chosen for IMAP can be divided into two broad categories: physical/chemical and biological. Physical/chemical indicators chosen for

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IMAP include the traditional water quality measurements such as temperature, salinity, dissolved oxygen, and pH, in addition to nutrients and chlorophyll a, which are indicators of productivity. Most water quality measurements will be obtained using an automated sampler (Hydrolab®). Seagrass community composition will be assessed with two methods: 1) a visual examination of % cover occupied by each species, and 2) a core sample in which seagrasses are returned to the lab for detailed analyses. Submerged Aquatic Vegetation Analysis

This presentation will describe the rationale behind using submerged aquatic vegetation (S.A.V.) as an environmental indicator, as well as the equipment and processes involved in the assessment of seagrass communities. Methodology explanation will include site selection and targeting, measurement of light intensity above and below the water surface, water quality assessment, species identification, qualitative and quantitative enumeration of S.A.V., and collection of seagrass core samples.

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The Ph.D. Program in Coastal Resource Management at East Carolina University

Russ Lewis East Carolina University

3904B Palmer Dr. Greenville, NC 27858

Russ.Lewis@lycos.com Abstract

The Ph.D. program in coastal resource management emphasizes an integrated, interdisciplinary approach to coastal studies with a focus on science and public policy. The program is designed to nurtures skills in the acquisition, interpretation, and synthesis of scientific information on coastal environments and populations. The structure of the program fosters pursuit of individual interests in the context of a structured, but flexible, program of classroom instruction, field research, work experience, and a doctoral dissertation. The program draws on a supportive, collaborative faculty from some 16 academic departments, plus close ties to marine science faculty at the University of North Carolina-Chapel Hill and Wilmington; North Carolina State University, and Duke University. The ultimate goal of the program is to provide the academic basis for students seeking resource management careers in government agencies, private firms, non-profit organizations, and interdisciplinary educational programs Areas of Study

Students may concentrate in one of four areas, with complementary work in two others. Coastal and Estuarine Ecology focuses on near-shore and estuarine processes that are important for living marine resources and environmental quality. Coastal Geosciences emphasizes coastal processes, geomorphology, and hydrology as they affect use and development of the coastal margin. Social Science and Coastal Policy investigates natural resource economics, politics and public policy, demographic trends and social behavior as they relate to coastal management. Maritime Studies explores the cultural and historical dimensions of coastal resources with a focus on maritime history, nautical archaeology, and the role of maritime heritage in coastal use and development.

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Considerations for Scientific Technical Diving: An Overview of Logistics, Procedures, and Implications for Program Development

Michael R. Lombardi* Founder/President

Applied Subsea Technologies Inc. (508) 725-8122

appliedsubtech@juno.com

*Interim Dive Safety Officer PIMS-Caribbean Marine Research Center

Lee Stocking Island, Exuma, Bahamas Abstract

Recent interests in conducting science at depths beyond traditionally accepted manned-diving operation depth limits has challenged diving scientists to embrace new and slowly emerging techniques in mixed-gas diving. For decades, the exploration in scientific pursuits has been lost in a world of institutional bureaucracy and politics, thus limiting the potential that our ocean has to offer. By adopting and applying the ‘tools’ (open- and closed- circuit mixed-gas SCUBA) developed by the industry, a new era of true exploration will drive a rejuvenated trend in discovery through science. The Caribbean Marine Research Center is taking steps to support a group of scientists in these pursuits.

“…we have an opportunity to set our sights on a much broader horizon. The time has

come to take exploration farther west, and east, and south, to our submerged continents. In so doing, we can challenge and rekindle American’s spirit of exploration…”

-2000, US President William J. Clinton Introduction

Over the past several years, interest has been generated in conducting exploratory science in deep/extreme environments. Most of these (i.e., deep walls, caves) are well beyond the reach of conventional SCUBA and require substantial equipment, training, and self-discipline to appropriately venture into these areas. Shallow reefs (<130 fsw) and other subaquatic environments have been the focus of marine science for several decades (CMRC scientists made over 3000 dives <130fsw in 2001) (Figure1). The organisms’ life histories and environmental processes affecting organisms at these shallow depths are very well documented.

Previously, much of the effort to explore and study the deep has been in using manned submersibles, ROV’s, AUV’s, etc. (CMRC supported 5 submersible dives from 500 to 900fsw in 2001). These studies typically focus on very extreme habitats such as

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thermal vent communities, ocean trenches, and deep blue water…all of which are out of reach by an individual diver exposed to ambient pressures at this point in time. These efforts are very expensive, averaging upwards of $10,000/day for a full suite of equipment and support personnel. Due to this expense, much of the mid-water depths (greater than 130 fsw, less than 500 fsw) have been overlooked, as they are not considered ‘elaborate’ or ‘exciting’ to scientists exploring extreme depths. Subs capable of working at thousands of feet do so ‘because they can’. This pattern (Figure 1) of several thousand shallow SCUBA dives and only a handful of submersible dives is common at most marine facilities and research institutions. This is a clear representation that we have only just scratched the surface of ocean exploration. The two depth extremes commonly sought out by scientists have left us with the challenge of determining the most effective means of not only accessing, but also efficiently conducting science in intermediate depths.

Recent advancements (last 20 years) in open-circuit and closed-circuit SCUBA training and equipment, do provide the highly skilled scientist an affordable first hand look at these deep(er) communities for on the order of hundreds of dollars per day. However, the regular application of these technologies has not been accomplished. Actually placing divers at these depths allows for macro- and micro-scale surveys, and careful manipulation and collection of specimens; tasks that are not easily accomplished by large and awkward manipulator devices on submersibles. The maneuverability of an individual diver in a manned deep diving operation offers unparalleled capability to conduct science at every spatial scale.

Very few groups have conducted science at these depths, as there are several issues to be dealt with, especially with regard to diving program standards, procedures, and liability concerns. Several private sector exploration groups do conduct and support science (e.g. GUE/WKPP, USDCT’s Wakulla 2 Project, Cambrian Foundation, etc), however there are standing liability issues for federal and state funded institutionally-based diving beyond the current maximum AAUS’ depth limit of 190fsw. Rich Pyle (1992) is one individual who has paved the way for deep exploratory science with his taxonomic work on reef fishes in Hawaii and throughout the Pacific. His work over the past decade has proven that we possess the technology and knowledge to safely venture into new areas for scientific pursuits. At present, AAUS has no formal standards in place for advanced diving modes (mixed-gas, rebreathers, caves, etc), although each organizational member likely has an ‘emerging technologies’ clause in place which allows these activities under the discretion of the home institutions’ Diving Safety Officer and Diving Control Board.

Although challenging from several viewpoints, mixed-gas SCUBA is a logical first step to pursue science in these intermediate depths. Defining routine capabilities to access these mid-water depths will remain an ever-present challenge, as there clearly must be an expressed need to support these developments, however significant efforts have been recently made on a program-specific basis. With the interest of some scientists lying in deep reef environments, the Caribbean Marine Research Center (CMRC)

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recently addressed this issue and is taking a bold step to pursue dive program advancement and become a leader for supporting science in deep/extreme habitats. May 2002 technical diving program

In May 2002, CMRC hosted its first mixed-gas program to support science objectives to depths approaching 300 fsw. The program was broken down into 2 distinct phases. Phase 1 included an intensive 8 day IANTD training program. Phase 2 included a series of deep working dives to support the work of Drs. Michael P. Lesser (University of New Hampshire) and Marc Slattery (University of Mississippi) (Figure 2).

Going into the program, several goals were kept in mind for this project with particular attention towards implications for future work. These included:

1. assess the feasibility of conducting mixed-gas decompression dives at this remote site

2. establish and uphold an acceptable set of standards for this advanced mode of diving

3. successfully obtain workable data during science dives to pave the way for future deep exploratory science programs

4. determine the potential for CMRC becoming a formalized training ground for these activities within the scientific community

5. promote future applications of advanced scientific diving techniques Logistics Operations manual

The logistics involved in organizing a program of this magnitude at a remote location proved to be very challenging. Step one was to establish a training and working dive protocol to serve as program guidelines. Several months prior to the start of the project, the team collaborated to compile a draft of a standards and procedures manual for mixed-gas decompression diving activities. This was modeled very closely after the NOAA-NURC program in place at UNC-Wilmington which has been using advanced diving modes for several years, specifically working on the USS Monitor Project. Ops manual content covered everything from training and personnel roles, to on-site procedures and every aspect of safety and accident management. Once finalized, the manual was submitted for final review and approval by CMRC’s Diving Control Board. Project DSO Michael Lombardi made all final revisions based on DCB input and formatted the document as it is in its present state (2002). Safety

Prior to training and work activities, the Project DSO held safety meetings to review CMRC emergency protocols and to define personnel responsibilities. Emergency procedures included notifying the two closest hyperbaric chamber facilities (Miami and

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Nassau) and the USCG of our deep diving plan. Evacuation procedures were covered, and CMRC staff had a secondary internal meeting to reinforce and refresh emergency protocol. A detailed outline of CMRC emergency evacuation procedures and emergency phone numbers were readily available to all staff during the project, and were also carried by surface support on-site.

Concerns for on-site contingencies were discussed and included omitted decompression, symptomatic in-water DCS, oxygen toxicity, and being chased out of the water by sharks. Being at a remote location, there were discussions regarding in-water recompression as an alternative to a timely evacuation for mild symptomatic DCS. Presently, CMRC’s standard evacuation procedures remain in place for any symptomatic DCS incidents, regardless of whether or not they occur during normal dive ops or during staged decompression diving activities.

Dive site safety included use of 2 vessels. One primary and one secondary which served as a chase boat in case of drifting decompression or the need to make an emergency run. Each boat was equipped with an oxygen kit and at least one boat was equipped with a GPS. Radio checks were made at key points during the 4.33nm drive to the work site. A final radio check was made once the divers entered the water.

Several days before the project, two rope highways were laid out from the base of the permanent on-site mooring (~80fsw) to approximately 400fsw to provide a reference point for the dive teams. These rope highways blocked off an area such that divers could easily locate either line and use it as a direct link to their decompression cylinders and back to the surface. Additionally, two uplines were affixed to the bottom to provide a stable (i.e. no vertical movement) decompression platform. Equipment requirements

Gas (6, 337 ft3 bottles each of He and O2) was ordered 5 weeks in advance of the project. Gas was shipped to the Bahamas via barge, and eventually delivered to CMRC via truck and boat. CMRC has nitrox capabilities in place via a UBS membrane system. EANx40 was stored in large bank bottles to use during partial pressure filling in an effort to save oxygen. At this point in time, CMRC does not have a booster pump in place to maximize gas filling efficiency. Air quality tests indicated that our compressors were pumping air suitable for partial pressure blending, so no tertiary external hyperfiltration systems were used. Four oxygen analyzers were used during the project, all of which had new sensors installed.

Several cylinders were dedicated for technical diving use. All were oxygen serviced according to NOAA’s oxygen service guidelines and properly labeled for trimix, nitrox, or high FO2 decompression gases. CMRC presently has 2 sets of twin AL80’s and 1 set of twin LP AA95’s, all with DIN/K capable isolation manifolds. CMRC also has 6 dedicated nitrox decompression cylinders (AL80’s) and 6 dedicated high FO2 decompression cylinders (AL80’s), all also with DIN/K capability. This equipment in

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place is adequate for supporting research teams in deep and/or cave environments according to CMRC’s proposed standards.

All project divers provided their own personal diving equipment. Estimated individual cost for technical equipment is approximately $2500-3000. Detailed requirements (Lombardi ed. 2002) were issued prior to the project to ensure that the concept of ‘team compatibly’ was satisfactorily met. The team dove a fundamental hogarthian configuration, of course emphasizing streamlining and redundancy. Divers breathed the long hose (7’) off of the right post to conform with the current configuration theories in the dive industry. Fundamental configuration included a set of doubles with bottom mix, a wings/harness style BC, two decompression cylinders, and an assortment of safety items (cutting-devices, slates, reels, lift bags, etc). Each diver was required to gain familiarity with other team members specific units during training. Training

Brian Kakuk, present DSO at CMRC, conducted the IANTD course training. There was an emphasis on mission oriented dives and dive planning. The training consisted of several hours of confined water work, and a series of 8 dives using various combinations of trimix, nitrox, air, and oxygen to depths approaching 270 fsw. The classroom work for the course lasted 3-4 hours per day and thoroughly covered every aspect of technical diving. There was also informal training in gas blending, as that was likely the most time consuming portion of the project.

During confined water sessions, divers were required to share air in various circumstances, ditch and don the full technical equipment configuration underwater and at the surface, identify and isolate various valve/cylinder failure scenarios, manage decompression cylinders, and remove an injured diver from their gear. Once all divers were comfortable with those skills, several skills were repeated with the divers being blindfolded. This greatly increased awareness of personal equipment placement and how to assess and cope with difficulties in the worst possible scenario. During every training dive, divers were drilled on sharing air, decompression cylinder management, isolating various manifold/regulator failures, and lift bag deployment to simulate drifting decompression.

Procedures Personnel

During training, IANTD standards were followed with respect to personnel requirements. However, the approved CMRC operations manual called for a 7-man dive team on-site to support actual working scientific dives. This team-size was modeled after NOAA dive team requirements (NOAA Dive Manual 4th Ed.). These roles were broken down as follows:

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Research divers (2)- Equipped appropriately for diving to target depth, carrying all decompression gas to personally complete the dive, and carrying appropriate scientific equipment.

Standby diver (1)- Equipped to make a solo dive to target depth in response to an

emergency. Support divers (2)- One would dive with a technical equipment configuration and carry

extra decompression cylinders. The second would use standard SCUBA and was responsible for relieving the deep team of extraneous science equipment. Support divers were restricted to 110fsw and breathed air throughout the dive.

Vessel operator/dive supervisor (1)- Assisted topside and was responsible for completing

a dive plan worksheet for each training and work dive. Vessel operator/chase boat (1)- Assisted topside and coordinated deployment of support

divers. Also responsible for keeping an eye out for and following deployed lift-bags, potentially indicating an emergency.

Dive Limits Oxygen exposure limits- Previous CMRC oxygen exposure limits for nitrox diving include a PO2 of 1.4 ATA. In order to optimize decompression, the technical operations manual was written such that a PO2 of 1.4 ATA was the accepted maximum for the bottom exposure, with a contingency PO2 of 1.5 ATA. A PO2 of 1.6 ATA was acceptable for decompression. Although not generally a concern for short duration deep dives, pulmonary O2 toxicity was tracked using the standard OTU/REPEX method. Equivalent narcotic depths- An END of 120fsw was selected as the target maximum. This afforded the deep team with an extremely narcosis-free head while working at 300fsw. Gas selection and management- Gas selection for the 300fsw trimix dives was highly dependant on ease of blending various mixtures without a booster pump. For decompression, EANx40 was used, as we had an unlimited supply via the UBS membrane system. EANx75 was used as our second decompression gas, as this was also easily obtained using pure O2 with an EANx top-off. Trimix 14/50 was used for the bottom mix, as it met our requirements for a max PO2 of 1.4 ATA and an END of 120fsw at our maximum target working depth. Gas management included the rule-of-thirds for bottom mix, and the required volume of decompression gas was multiplied by a conservative factor of 1.2. This typically resulted in divers using twin AL80’s at a minimum for bottom mix and AL80’s for decompression cylinders, although AL30’s and AL40’s were used on some occasions. Based on the varying RMV’s of our project team, on a few occasions, travel cylinders were also carried throughout the dive and used to supplement the bottom mix.

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Decompression- Bottom times were limited to 15 minutes, including descent, for the scientific working dives. This was in an effort to minimize the decompression obligation, and in the event of a lost decompression cylinder, a reasonable profile could be resumed and appropriately carried-out using bottom mix, the remaining decompression gas, or an extra cylinder from a support diver. The short dive time eliminated the need to stage various gases at a number of depths at the work site. Bottom gas consumption would also become an issue if much more than 15 minutes were considered for work. Likewise, a maximum depth of 300fsw was arbitrarily selected. This depth seemed reasonable based on the varying experience of the project team, the complications in prepping for any dive much deeper, and this provided our group with access to more than twice the depth normally worked by scientists at this site. A fair amount of time was spent discussing contingencies, everything from equipment failure to aborted deco schedules or altered dive plans. Decompression schedules were determined using IANTD tables (Buhlmann Zhl16 algorithm) and were cross checked with decompression software (Proplanner, Dr. X, and Abyss were all used). Safety factors incorporated into the dive profile included a 2 minute deep stop at 170fsw, an additional 5 minutes in-water time at the 20fsw stop breathing EANx75, and five additional minutes were spent at the surface breathing EANx75. Total runtime for these dives was typically 100-120 minutes, depending on ascent rates and travel times between stops (Figure 3). An ascent rate of 60fpm was maintained until reaching the deep stop, followed by an ascent rate of no more than 30fpm for the remainder of the profile. ***These imposed limits on the dive operation did prove effective and were satisfactorily met. It is difficult to formally impose strict limits, as each dive project or objective differs in suitable dive plans, hence some factors vary slightly. In supporting future technical diving at CMRC, each project will likely have to submit a detailed dive plan, incorporating appropriate O2 exposure, gas management, and decompression scenarios which will then be subject to approval by the DSO and DCB. ***Although trimix diving affords divers with relatively simple and inexpensive access to depths beyond conventional air diving, the 15 minute bottom time is a cause for concern in executing considerable amounts of science. Both extended decompression obligations and gas management issues must be dealt with for much longer exposures at these depths. Closed-circuit rebreathers would alleviate these problems almost entirely, by providing a near unlimited gas supply, and maximizing efficiency of decompression by offsetting inert-gas uptake with a constant optimized PO2. Science Dives

Bock Wall (23˚ 49.920’ N 076˚ 09.158’ W) was selected as the primary research site, as it has been the target of several studies by this research team in the past. Two days of science dives were conducted to 300 fsw. Once each vessel arrived on-site, the Project DSO gave a safety briefing to finalize the dive plan and science objectives.

During descent, the divers breathed EANx40 as a travel gas to 80 fsw, at which point they switched to bottom mix (trimix 14/50). Divers also dropped off their ‘hot’ mix of EANx75 at the base of the site mooring. At 110 fsw, the nitrox cylinders were also

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clipped off. Divers made one final check, then commenced with descent to 300 fsw. 15 minutes were spent at depth collecting specimens, water samples, and shooting video (during the course of 2 dives). Divers then began their ascent and decompression process (Figure 4).

At 110 fsw during ascent, the deep team was met by two support divers. One was equipped in full technical gear and carried extra cylinders of EANx40 and EANx75 in case of a failure by either member of the deep team. The second support diver was equipped in standard SCUBA attire and was responsible for relieving the deep team of any extraneous science equipment. All four divers followed out the deep team’s decompression schedule. Support divers remained breathing air; however they were required to stay in-water to monitor the deep team for signs of oxygen toxicity, or in-water DCS. The Science

In accessing any new environment, several big-picture questions must be considered. This includes the biodiversity of the system, the role that this system plays in contributing to the function of the entire reef community, identifying new species, and further determining which new species may offer exploitable resources, and then ultimately preserving those resources. Having access to these depths clearly open the doors to the next century of marine science, and opportunities abound to unveil several significant discoveries. With respect to actual science, aside from being absolutely awe-struck from our first look at the benthic community (LOTS of work to do!), we were able to gain valuable information from this series of trimix dives. During one 300fsw dive (~8 minutes of workable dive time), 29 species of sponge were collected. Several of these posed problems in taxonomic identification, and others (~55%) were previously only known to be found in nearby marine cave systems. These species are of particular interest in the natural products/biomedical research field, as they leach antimicrobial compounds, being investigated for potential use in combating HIV, diabetes, cancer, among other diseases. Implications for future work

The work described above represents only a small first step towards applying advanced manned diving capabilities to science, and attempting to gain a better understanding of marine habitats in mid-water depths. Trimix affords the scientist with reduced narcosis, and reduced PO2 exposures, thus allowing ventures to depths beyond air diving limits with relative safety. However, this has presented a certain number of clear limitations and complications. Task loading associated with conducting science on top of carrying out a very specific dive profile is difficult. Also, the time restriction imposed on us by both decompression obligation and from a gas management standpoint greatly influences the amount of science we can effectively conduct.

With trimix or heliox being the obvious gas choice, the mode for its application may be arguable. While open-circuit mixed-gas diving has its place, the cost per dive is

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relatively high based on gas consumed for the amount of allowable bottom ‘working’ time. For extended durations, or further pushes of depth, closed circuit rebreathers are obviously the mode of choice. CCR’s use very little gas in comparison to the amount of time spent conducting science (Parrish and Pyle 2002). CCR’s also maximize decompression efficiency by maintaining a constant PO2, thus reducing inert gas partial pressures as compared to open circuit diving. Our research team is beginning to assess the appropriateness of incorporating CCR’s to conduct science more efficiently at mid-water depths approaching and eventually beyond 300fsw. Conclusion

We have only just broken the surface in exploring our oceans in their entirety. Due to various limits imposed on us as diving scientists, access to deeper unexplored areas is extremely challenging. Despite several significant exploratory achievements over the past two decades, application of mixed-gas diving is still only in its infancy, especially in the scientific community. It is difficult to determine how routine these types of dives will become, as forward progress in developing technology, procedures, and standards is still very much in the hands of pioneering divers and scientists who are ‘thinking outside the box’. For decades, development and application of diving technology has been the result of a demonstrated need from the scientific community. There is certainly a ‘science of diving and diving technology’, and its development over the next several years should be viewed as an opportunity for the science community to embrace new technologies as a tool to answer new questions. One might ask how we can successfully put a man on the moon, yet we don’t have near regular access to 97% of the ‘innerspace’ on our own planet. These mid-water depths are critically important in contributing to the functionality of our oceans and our planet, and we have fortunately taken the first step and just begun to investigate these areas. Extending humans’ limits from a technology standpoint, as well as in our pursuits of scientific truths are crucial for the future of ocean exploration by government, military, commercial, and scientific organizations. Some of the greatest discoveries of the 21st century likely lie in our ocean, and by further developing innovative diving apparatus and techniques, implementing new programs, and redefining our limits, we can begin to determine the best approach to routinely examine our ‘innerspace’. Mixed-gas technical diving, both open- and closed- circuit, is only the first step.

…to anyone who does not dare envision our abilities to progress beyond today’s limitations to a new tomorrow, I declare, ‘never say impossible’.

-Mr. John H. Perry Jr.

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Project Dive Statistics The following table summarizes 2002 CMRC technical dive training activities: Instructor-Brian Kakuk Students-Michael Lesser, Liz Kintzing, Marc Slattery, Michael Lombardi Support-Erin Rechisky, Kent Reinhold, Kevin Buch, Mette Blaesbjerg dive# date gases used max depth bottom time runtime #divers 1 5/17 air, N75 152 30 74 5 2 5/18 air, N40 181 25 91 5 3 5/19 air, N40 185 25 91 5 4 5/19 air, N40 130 20 60 5 5 5/20 TX16/40,N36, 237 20 97 5 N75 6 5/21 TX1640,N40 269 20 109 5 N77 7 5/22 TX19/12,N40 160 15 49 5 8 5/22 TX19/12,N40 102 15 39 5 total # person dives: 40 cumulative bottom time (5 divers) 850 min (14.2 hours) cumulative runtime (5 divers) 3050 min (50.8 hours) Gases used: air 20 sets of doubles (AL80’s, AA95’s, HP AA100’s) 4 single AL80’s

N75(77) 15 cylinders (AL80’s, AL30’s) N36 5 cylinders (AL 80’s) N40 25 cylinders (AL80’s, AL40’s) TX19/12 5 sets of doubles (AL80’s, AA95’s, HP AA 100’s) TX16/40 10 sets of doubles (AL80’s, AA95’s, HP AA 100’s)

Estimated time spent preparing gas: 4-5 hours/dive (~40 hours total)

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The following table summarizes 2002 CMRC scientific technical dives: Project DSO-Michael Lombardi Research divers-Michael Lesser, Marc Slattery, Michael Lombardi Standby divers-Michael Lesser, Marc Slattery Support divers-Kent Reinhold, Kevin Buch, Caleb Thibeault Vessel Operators-Erin Rechisky, Mette Blaesbjerg dive# date gases used max depth bottom time runtime divers 1 5/27 TX14/50 302 15 109 Lombardi N40,N75 Lesser Support 1 air 110 5 76 Buch Thibeault 2 5/28 TX14/50 308 15 97 Lombardi N40,N75 Slattery Support 2 air 110 5 81 Reinhold Thibeault Total # person deep dives: 4 Cumulative bottom time (3 divers) 60 min (1 hour) Cumulative runtime (3 divers) 412 min (6.9 hours) Total # person support dives: 4 Cumulative bottom time (3 divers) 20 min Cumulative runtime (3 divers) 314 min (5.23 hours) Gas used: Air 2 sets of doubles (AL80’s) 4 single AL80’s N70 7 cylinders (6 AL80’s, 1 AL40) N40 7 cylinders (6 AL80’s, 1 AL30) TX14/50 5 sets of doubles (2 AL80’s, 2 AA95’s, 1 HP AA100) Estimated time spent preparing gas: 4-5 hours/dive (~10 hours total) ------------------------------------------------------------------------------------------------------------ In demonstrating the capabilities and effectiveness of this training/science program, the following is a brief summary of dives made at CMRC deeper than 130 fsw in the past 3 years: 2001 11 dives 338 min total 157 fsw max D 2000 41 dives 1689 min total 160 fsw max D 1999 1 dive 92 min total 130 fsw max D total dives in past 3 years: 53 total dive time: 2119 min (36.3 hours) During the weeklong program in May 2002, the science and support group spent more in-water time during dives deeper than 130fsw than all totaled deep dives for the past 3 years at CMRC.

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Acknowledgements

The author would like to thank the members of the 2002 project team; Michael Lesser, Marc Slattery, Liz Kintzing, and Brian Kakuk. Also, thanks to the support group of Erin Rechisky, Kevin Buch, Mette Blaesbjerg, Kent Reinhold, and Caleb Thibeault. CMRC provided continuous professional support during all aspects of the program. Thanks to Odyssey Scuba for assistance with equipment. Finally, the entire group extends many thanks to John Marr, center director CMRC, for his continued support in advancing scientific diving and of our science objectives. Both the training and science components of this project were supported by a CMRC Program Development Grant to M. Lesser (Using Mixed Gas Technical Diving to Study the Ecology of Deep Water Sponge Communities, CMRC-02-PRML-03-02A).

The author can be contacted for information regarding technical diving programs and training inquiries.

The views expressed herein are exclusively those of the author and do not necessarily reflect the views of NOAA, NURP, PIMS-CMRC, their officers, employees, or project personnel. References Gilliam, B. 1995. Deep Diving. An Advanced Guide to Physiology, Procedures, and

Systems. San Diego, CA: Watersport Publishing Inc. 352pp. Lombardi, M.R., editor 2002. The next step: Mixed-gas Technical Diving ‘Science 300

feet deep’. Procedures for diving operations using mixed-gas, open-circuit SCUBA with staged, in-water decompression for conducting undersea science exploration. NOAA-National Undersea Research Program-Caribbean Marine Research Center internal document.

NOAA Diving Manual 4th Edition. 2002. James T. Joiner, Ed. US Department of

Commerce, National Oceanic and Atmospheric Administration. Parrish, FA, and Pyle, R. 2002. Field Comparison of Open-Circuit Scuba to Closed-

Circuit Mixed-Gas Diving Operations. Journal of the Marine Technology Society. Vol. 36, No. 2.

Pyle, R., and Sharkey, P. 1992. The Twilight Zone: The potential, problems, and theory

behind using mixed gas, surface-based SCUBA for research diving between 200 and 500 feet. In ‘Diving for Science 1992’ AAUS annual symposium proceedings. Pp173-187.

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Figures

0

0 to 130

130 to 300

130 to 300

300 to 500

500+

Dep

ths

(fsw

)

Figure 1. Schematic o<130fsw. Only 11 wersubmersible dives werconducted deeper thanclear area of our ocean

2001-11 dives, 157fsw max (Lesser/Lombardi)

2002-44 dives, 308fsw max (Slattery/Lombardi) 5 sub dives (Powell)

500 1000 1500 2000 2500 3000 3500

# Dives

f 2001 CMRC scientific dives. More than 3000 dives were made e made deeper than 130fsw, with a max D of 157fsw. 5 e made in 2001, all deeper than 300fsw. In 2002, 44 dives were 130fsw, with a max D of 308fsw. This is progress, yet there is a that has not been thoroughly explored.

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Figure 2. CMRC’s 2002 trimix program team (left to right): Marc Slattery (University of Mississippi), Michael Lesser (University of New Hampshire), Michael Lombardi (Applied Subsea Technologies), Brian Kakuk (Caribbean Marine Research Center), and (front) Liz Kintzing (University of New Hampshire).

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Figure 3.target depappropria

Runtime (min)

0

50

100

150

200

250

300

0 20 40 60 80 100 120

Dep

th (F

SW)

Gases Used: EANx40 travel to 80FSW TX14/50 bottom mix EANx40 deco from 110FSW EANx75 deco from 30FSW

300fsw mixed-gas profile for working dives. 15 minutes were spent at the th ‘working’, which required a total runtime of over 100 minutes after making te gas switches to optimize decompression.

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Figure 4. M. Lombardi (left) and M. Lesser (right) decompress after a 300fsw scientific dive.

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Using Diving and Remote Sensing Approach to Classify Benthic Habitats in Coral Reef Ecosystems in Belize

Joseph J. Luczkovich1,2

Jason W. Rueter1

1Department of Biology and 2Institute for Coastal and Marine Resources East Carolina University

Greenville, NC, USA 27858 luczkovichj@mail.ecu.edu

Abstract

We examined the use of multivariate cluster and discriminant analysis of ground truth data and LANDSAT 7 satellite imagery to classify tropical marine habitats at Turneffe Atoll, Belize, Central America. Ground truth data were obtained using SCUBA, digital video and GPS. Ground-truthing was done by swimming along eleven 150 to 300-meter transect lines located on the southeastern side of the atoll and videotaping bottom habitat from 1.5 m above the bottom at 30-meter intervals (126 sites). Still images were taken from the video for each location, overlaid with a 20-cell grid, and % cover of seven bottom habitat classes was computed. Using hierarchical cluster analysis of water depth and habitat % cover data, 78 of the 126 sites were grouped into three distinct habitat clusters (sand, seagrass, and coral reef habitats). A discriminant analysis of the sites was performed using LANDSAT digital values in TM Bands 1, 2, 3, and 4 (the only ones that penetrate water) as predictors of habitat cluster membership. Coral reef areas were correctly classified 100%, seagrass areas 80 %, and sand-dominated areas 77 % of the time with our training set; we will need to further validate this approach by ground truthing predictions of the discriminant function. Using the classification function from the discriminant analysis, we have recoded the LANDSAT 7 image to create a habitat map of the region surrounding Turneffe Atoll, showing coral reef, seagrass, and sand-dominated regions. Such habitat maps can be used to study change in habitat area over time.

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Noisy Fish and even Louder Divers: Recording Fish Sounds Underwater, with some Problems and Solutions using Hydrophones, Sonobuoys, Divers, Underwater Video and ROVs

Joseph J. Luczkovich1

Mark W. Sprague2

1Institute for Coastal and Marine Resources and Department of Biology 2Department of Physics

East Carolina University, Greenville, NC 27858 luczkovichj@mail.ecu.edu

Abstract The sea is a noisy place. Over 400 species of fishes produce species-specific sounds in response to disturbance, during agonistic encounters in defense of territories, and in spawning events. It is desirable to use the sounds produced by fishes in the study of their behavior and ecology and fisheries management; this is called the passive acoustic approach. The first step in the use of passive acoustics is to identify which species of fish makes a particular sound, and under which conditions such sounds are made. In the past, fishes have been recorded in captivity to obtain the “sound-truth” data; this provides an unequivocal and inexpensive result, but it has some problems – e.g., tank echoes, behavioral inhibition in artificial environments, and inability to study large fishes (e.g., marlins that may make sound). An obvious alternative is to use SCUBA diving or ROVs to approach and record fishes in natural environments. But divers and underwater vehicles create their own sound environments that mask or interfere with fish acoustic recording. In addition, determining the location of sound sources in acoustically complex ocean environments can present its own set of problems. We will discuss these problems and how they can be overcome. We will present technologies and approaches that have been used by our Fish Acoustics Research Team to record fishes in situ – hydrophones suspended from vessels, autonomous sonobuoys, underwater video recorded along with sounds using calibrated hydrophones, either diver-deployed or on an ROV. Using these methods, we have been able to identify and record fishes in the drum family (Sciaenidae), cusk-eel family (Ophiididae), and toadfish family (Batrachichoididae) in North Carolina and damselfish (Pomacentridae) and squirrelfish family (Holocentridae) on coral reefs in Belize, among many others. A sonobuoy that recorded on a timer 90 s every hour for 24 h was used successfully to study sound production in multiple locations over time. Sound production in the many of these fishes was greatest after sunset, due to increased activity of these fish and nocturnal mating behavior. Behavioral acoustic avoidance interaction of silver perch (Bairdiella chrysoura) occurred when bottlenose dolphin whistles were recorded in the area or played back experimentally. Misidentifications have occurred in the past and unknown sounds of biological origin have been detected on recordings that do not match any of the captive species recorded, so further work underwater will be required to positively identify such soniferous species. A fish sound catalogue from captive and in situ individuals is being collected for use in the

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identification of unknown sounds and to monitor the condition of reefs and ocean spawning areas remotely. Rebreather technology can provide an alternative to SCUBA for making in situ recordings.

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Hampster Balls, Voyeurism, and Video: A Method of Monitoring Reproductive Behavior in Coral Reef Fish

Sean J. Lyman Center for Hyperbaric Medicine and Environmental Physiology

Box 3823 Duke University Medical Center

Durham, NC 27710 Sjl3@duke.edu

Will. F. Figueira

Duke University Marine Laboratory 135 Duke Marine Lab Rd.

Beaufort, NC 28516 Abstract

In situ studies of fish behavior can be challenging. The expense and limited bottom time of underwater research make each observation precious, while the three-dimensional environment of a fish precludes the use of many terrestrial experimental techniques. For these reasons, many in situ behavioral studies have been just that—observational studies, rather than manipulative experiments. To overcome these difficulties, we developed a method of containing and presenting potential mates and predators to male damselfish and utilized underwater digital video to capture the reaction of the study fish. The requirements of the study were: a means to contain predators and potential mates in a way that visual and chemical cues would be undisturbed and enough fresh water would flow through the container to keep the fish alive; a means of presenting the fish to the subject male without the diver approaching the subject too closely; and, a means of recording subject reaction to the presented fish. The solution we developed was to place the fish in modified clear hamster exercise balls that could be attached to a long PVC pole, and to use video to monitor subject reaction. These video observations were backed up by diver observations, and the effect of diver and video camera on fish behavior was attenuated by an acclimation period. Save for a single unpleasant barracuda interaction, the method was successful in allowing for a manipulative experiment investigating the tradeoff between mating and risk of predation in a coral reef damselfish.

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Scientific Diving Training through an Aquarium Diving Program

Peter Pehl II Aquarium of the Pacific

100 Aquarium Way Long Beach, CA 90802

Ppehl@lbaop.org Abstract Aquaria such as the Aquarium of the Pacific in Long Beach California maintain an active and experienced diver force amongst the staff and volunteers. With scientific diving training that meets the standards of AAUS, these divers are an asset that can be readily utilized to support university research programs. A description and discussion of the AoP Scientific Diver Training course will be co-presented by Peter Pehl II, AoP Diving safety Officer and Professor Roy Houston of Loyola Marymount University.

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Rubber Duckies or Shipwrecks: A Submerged Cultural Resource Survey of Bath, North Carolina

Andrew Pietruszka Maritime Studies Program

East Carolina University 1704 E. 3rd St.

Greenville, NC 27858 Apt0823@mail.ecu.edu

Abstract

Bath, North Carolina is the oldest incorporated town in the state. It was incorporated in 1705 and soon there after became one of the five ports of entry for the colony. The Bath Creek Submerged Cultural Resource Survey was established to assess the potential of Bath Creek for holding cultural resources considered historically valuable. The survey is a three-part project consisting of a historical survey, remote sensing survey, and a planned ground truthing dive survey.

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Transthoracic Echocardiography (TTE) - A Tool to Monitor Unsafe Decompression Stress

Neal W. Pollock, Ph.D. Center for Hyperbaric Medicine and Environmental Physiology

Department of Anesthesiology Duke University Medical Center

Durham, NC 2710 Abstract

Decompression sickness (DCS) is associated with gas emboli (bubble) formation. While the lung is an effective bubble filter, several avenues exist for left ventricular gas emboli (LVGE) to arise. Since LVGE are believed to produce an elevated risk of neurological DCS, it is prudent to react conservatively if any are observed, regardless of symptom status. Two-dimensional transthoracic echocardiography (TTE) provides a technique to visualize circulating bubbles. TTE is being used to monitor the left heart during altitude decompression studies at a growing number of facilities. A training curriculum has been established at Duke University to credential technicians working with the devices. Background

The etiology of neurological decompression sickness (DCS) is uncertain. Although it is associated with gas emboli (bubble) formation, the manner in which bubbles arise is not fully understood. Mechanisms may include autochthonous (in situ) formation in central nervous tissue or translocation of venous gas emboli (VGE) to the arterial path. Translocation may occur through intracardiac shunts, across the pulmonary capillary bed (transpulmonary passage), or via any of the variable venous-arterial anastomoses.

The lungs are normally very efficient at filtering gas bubbles from the blood. Intracardiac shunts, or short cuts, however, can allow some blood, and any bubbles present at that time, to bypass pulmonary filtration and be disseminated throughout the body. There are various types and degrees of intracardiac shunts. Atrial septal defects (ASD) and ventricular septal defects (VSD) provide direct communication between right and left atria and ventricles, respectively. The direction of flow depends on the pressure gradient between the respective heart chambers. The gradient is from left to right for the most part and blood flow through shunts therefore usually follows this path. There are, however, conditions in which shunting can be reversed. In the case of a large ASD, this can occur within a normal breathing cycle. Transient reversals may be promoted by immersion, strain-release techniques (e.g., Valsalva or anti-G maneuvers) and elevated pulmonary arterial or right atrial pressure. Large VSDs are usually identified in the fetus

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or neonate and usually require surgical repair to prevent complications. Unobserved VSDs are more likely limited to physiologically insignificant left-to-right leaks.

One common form of ASD is a remnant of the fetal circulation – the patent foramen ovale (PFO). The valve-like communication between the atria is normally closed after birth as left atrial pressure rises. Tissue growth normally seals the flap within the first 24 months. However, ~25% of the adult population have some residual opening (Hagen et al., 1984; Lynch et al, 1984). While the degree of patency is negligible in the majority of individuals, 5-10% of the adult population may have PFOs that are functionally important (i.e., allowing a physiologically meaningful amount of shunting) (Fisher et al., 1995). Functional (or physiological) patency is thereby distinguished from anatomical patency. Problematically, the presence of a functional PFO appears to increase the risk of neurological DCS in divers (Wilmshurst et al., 1986; Moon et al., 1989; Wilmshurst et al., 1989; Bove, 1998; Schwerzmann et al., 2001).

While it is possible to screen individuals for PFO and exclude them from decompression exposure, this is not done by diving organizations for several reasons: 1) the risk of DCS, with or without PFO, is very small; 2) PFO is not the only means of developing arterial bubbles or DCS; 3) the risk associated with PFO can be reduced by employing decompression practices which minimize bubble formation; and 4) there is a small but measurable risk associated with the test used to identify PFO (i.e., bubbles are injected into the venous circulation to visualize any subsequent cardiac crossover) (James, 1990). Implications for Decompression Research

Valid research requires a subject pool that is representative of the population at large. For decompression research, this precludes the exclusion of volunteers based on the presence of PFO despite the potential for elevated risk. The challenge is to be inclusive while protecting the long-term health of all participants.

The primary protection from laboratory-induced DCS is the ability to rapidly initiate treatment in response to symptom development. This facilitates quick resolution and minimal risk of complications or residual effects. The ability to evaluate significant decompression stress independent of symptoms offers another avenue for protection. While Doppler ultrasound has been used for some time to estimate global stress, more recent developments in echocardiography are allowing more directed monitoring of potentially high-risk conditions. Ultrasound and Echocardiography

Doppler ultrasound was first observed to detect circulating gas bubbles more than 40 years ago (Franklin et al., 1961). Since that time it has become the principle tool for monitoring decompression stress outside of symptom development. The greatest

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strengths of Doppler assessment are that it is non-invasive and can be quick to complete. The chief limitation is that the relationship between observed gas emboli and symptomatic DCS is not consistently strong. While the correlation can likely be improved by methodological changes, most decompression studies continue to rely on symptom development as the study end-point.

Echocardiography combines ultrasound imaging and Doppler flow detection technology. The procedures rely on transducers that transform energy from one form into another. Ultrasound transducers contain piezoelectric elements that emit sound waves (acoustic energy) when excited electrically. Sound waves transmitted through the body generate echoes when changes in density are encountered. Echoes return to the transducer-receiver and the acoustic energy is transformed back into electrical energy, processed, and displayed graphically. 'Gain' controls adjust the amplification of returning echoes to optimize the displayed image. Originally, transducers transmitted and received the same frequency. More recently, transducers have been developed to transmit at one frequency and receive at another frequency to reduce interference. The transmitted frequency is known as the fundamental or first harmonic frequency. The received frequency (second harmonic) is a whole number multiple of the fundamental frequency. For example, a device may employ 2 MHz fundamental (transmit) and 4 MHz second harmonic (receive) frequencies. This 'harmonic processing' improves image resolution and is generally employed as a standard when available.

Two-dimensional transthoracic echocardiographic (TTE) imaging can be used to visualize the heart in cross-sectional view. This allows simultaneous visualization of multiple heart chambers and major vessels. Doppler monitoring, on the other hand, is generally restricted to the evaluation of a single major vessel or heart chamber. While TTE can be used to quantify the gas emboli loads (Eftedal and Brubakk, 1997) in a manner similar to Doppler scoring (Spencer and Johanson, 1974; Kisman et al., 1978; Eatock and Nishi, 1986), its greatest strength may be in detecting gas emboli on the left side of the heart (i.e., left ventricular gas emboli or LVGE) that can be distributed systemically. Since LVGE are implicated as causative agents in neurological decompression sickness, this in an important capability. While the absolute risk of DCS is not known, the conservative approach during altitude decompression research trials is to treat the presence of LVGE, regardless of symptom status, as a test-termination criterion. TTE Equipment and Use in Decompression Studies

Traditional two-dimensional clinical echocardiographic imaging systems are bulky in size and prohibitively expensive for most non-clinical applications (~$200-250KUS). The Brooks Air Force research facility was one of the few to employ these devices in altitude decompression studies. Fortunately, recent advances in miniaturization and technology have resulted in the availability of portable systems that provide adequate resolution for LVGE monitoring at approximately one-tenth the cost of the standard clinical systems.

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The SonoSite SonoHeart Elite portable ultrasound device is currently being used

at Duke University and will soon be used at collaborating labs (Defence Research Development Canada [DRDC, formally the Defence and Civil Institute of Environmental Medicine] and the Johnson Space Center). The unit weighs 5.7 lbs and has physical dimensions that are smaller than many laptop computers (13.3 in x 7.6 in x 2.5 in). Harmonics processing is available as an option (with 2 MHz transmit and 4 MHz receive frequencies).

The SonoHeart has been tested and certified for use in hypobaric conditions (at altitude equivalents up to 30,000 ft) in a development program conducted by NASA and Duke University. Testing is underway to certify the unit for use at shallow decompression stop depths. Standard precautions ensure the safety of the device in the closed hypo-/hyper-baric chamber environment. These include removal of the battery and location of the AC power supply outside the chamber (a DC power cable is brought into the chamber through a wall penetrator). In addition, a minimum 5 L·min-1 flow of nitrogen is directed into the underside of the device housing to eliminate oxygen from the warmest area and reduce the risk of combustion.

Pilot trials employing the injection of agitated saline and/or commercial ultrasound contrast agent (Optison®) have provided compelling evidence of the device's ability to visualize LVGE. The SonoHeart is currently being used for decompression safety monitoring as part of flying after diving and high altitude decompression studies. The device has been used in 122 subject-exposures in the flying after diving study between May, 2002 and February, 2003. One case of mild, pain only decompression sickness has been observed in these trials. The presence of VGE, monitored with Doppler ultrasound, was documented in two other subjects (Grade 1 VGE on the Spencer 0-4 grade scale [Spencer and Johanson, 1974]). No LVGE or neurological symptoms have been observed in the study to date. Implications for Future Research

The use of TTE monitoring during decompression studies is likely to become a standard as a growing number of research laboratories adopt the procedures. It is anticipated that the presence of LVGE will continue to stand as a test-termination criterion for altitude exposure. The procedures for diving decompression are complicated by the fact that the stressor cannot be simply removed by returning the subject to ground pressure as it works for altitude decompression. The conservative approach for LVGE observed during or following diving decompression is prophylactic recompression. It will be important to learn more about LVGE frequency and the true risk relationship between LVGE and DCS to determine if prophylactic recompression is warranted. Training Requirements for Local Credentialing

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Our initial studies relied on credentialed cardiac sonographers to provide training

and oversight of the monitoring program. Technicians inside the chamber established views and sent the signals outside for digital recording and projection. Sonographers located outside the chamber directed technicians and confirmed the presence or absence of gas emboli.

An in-house training program was developed once the monitoring protocols were established. The goal was to provide local credentialing of technicians trained to perform scans independent of sonographer oversight. The estimated training time is 34-40 hours depending on the background of the individual. The distribution of the training time is described in Table 1. Table 1. Estimated Time Required to Train TTE Technicians

Training Component Duration (hours)

Theory and Overview Orientation Lecture 1-2 Self-Study 1-6

Hands On Practice Small Group Training Sessions 10 Supervised Research Scans 15

Testing and Evaluation Written Competency Examination 2 Affirmation by credentialed cardiac sonographer 2

Continuing Education Round Table Recording Scan Reviews 3 Total 34-40

The orientation lecture covers ultrasound principles and equipment, cardiac anatomy, standard cardiac views and procedures, terminology and study protocols. Self-study relies on computer-based graphic reference material to improve visual orientation and identification skills. The time required for orientation and self-study varies with the experience of the trainee.

Hands-on practice involves both dedicated training sessions and research study experience. Trainees complete at least 10 separate, one-hour sessions of small group training. The maximum student to instructor ratio is 3:1 (either clinical sonographers or locally credentialed TTE technicians serve as instructors). Instructors direct student technicians verbally to improve imaging techniques and mastery of standard reference terminology. Students scan a minimum of three different subjects (employing all three standard views) per session.

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Student technicians deemed competent by small group instructors serve as intern technicians during research studies. A credentialed cardiac sonographer supervises intern activity. The supervisor may be physically located with the intern or present via radio/video links when scans are conducted in working pressure chambers. Interns serve as supervised research study technicians for a minimum of three complete research studies, monitoring a minimum of 12 different subjects with a minimum of three scans (one baseline and two trial scans).

The written examination covers cardiac anatomy, basic principles of ultrasonic monitoring, operating characteristics of the available devices, basic scanning techniques, and protocols to be followed during research studies. The criterion-based examination has a minimum passing score of 80%. Individuals failing to achieve passing grades are allowed to complete a revised examination in no less than 30 days.

Local credentialing is issued when the training requirements are met and the supervising sonographer affirms that the candidate can reliably establish appropriate views with acceptable speed and proficiency. Candidates must obtain a stable and clearly interpretable image via one of the three standardized views on a new subject within three minutes and follow this with a 90% continuously-interpretable 60 second recording including rest (30 seconds) and movement (30 seconds following three repetitive, maximal one-second isometric single limb contractions) phases.

Both credentialed and trainee technicians participate in continuing education activity to improve and/or reinforce competency and promote standardization of techniques. Recorded research scans from previous studies and/or training sessions are presented for group review and discussion. Credentialed technicians must participate in round table review sessions at least quarterly or as issues arise. Conclusions

The availability of relatively inexpensive, two-dimensional transthoracic echocardiographic devices which enable visual monitoring of the left heart can improve the safety of decompression studies by identifying LVGE before they are distributed systemically. Although we do not know the absolute risk, it is prudent to suspend altitude exposure if LVGE are observed, regardless of symptom status. Monitoring protocols and a training program have been established for decompression studies conducted at Duke University. Further research is required to determine the true risk relationship between LVGE and DCS. Literature Cited Bove AA. Risk of decompression sickness with patent foramen ovale. Undersea

Hyperbar Med 1998; 25(3): 175-178.

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Eatock BC, Nishi RY. Procedures for Doppler ultrasonic monitoring of divers for intravascular bubbles. Department of National Defence: Defence and Civil Institute of Environmental Medicine (DCIEM) Report No. 86-C-25, 1986, 28 pp.

Eftedal O, Brubakk AO. Agreement between training and untrained observers in grading

intravascular bubble signals in ultrasonic images. Undersea Hyperbar Med 1997; 24(4): 293-299.

Fisher DC, Fisher EA, Budd JH, Rosen SE, Goldman ME. The incidence of patent

foramen ovale in 1,000 consecutive patients. A contrast transesophageal echocardiography study. Chest 1995; 107(6): 1504-1509.

Franklin DL, Schlegel W, Rushmer RF. Blood flow measured by Doppler frequency shift

of back-scattered ultrasound. Sci 1961; 134(3478): 564-565. Hagen PT, Scholz DG, Edwards WD. Incidence and size of patent foramen ovale during

the first 10 decades of life: an autopsy study of 965 normal hearts. Mayo Clin Proc 1984; 59(1): 17-20.

James PB. Safety of contrast echocardiography in screening divers (letter). Lancet 1990;

336(8727): 1389-1390. Kisman KE, Masurel G, Guillerm R. Bubble evaluation code for Doppler ultrasonic

decompression data. Undersea Biomed Res 1978; 5(suppl 1): 28. Lynch JJ, Schuchard GH, Gross CM, Wann LS. Prevalence of right-to-left atrial

shunting in a healthy population: detection by Valsalva maneuver contrast echocardiography. Am J Cardiol 1984; 53(10): 1478-1480.

Moon RE, Camporesi EM, Kisslo JA. Patent foramen ovale and decompression sickness

in divers. Lancet 1989; II(8637): 513-514. Spencer MP, Johanson DC. Investigation of new principles for human decompression

schedules using the Doppler blood bubble detector. Office of Naval Research Tech Rep ONR Contract N00014-73-C-0094, 1974.

Schwerzmann M, Seller C, Lipp E, Guzman R, Lovbald KO, Kraus M, Kucher N.

Relation between directly detected patent foramen ovale and ischemic brain lesions in sport divers. Annals Internal Med 2001; 134(1): 21-24.

Wilmshurst PT, Byrne JC, Webb-Peploe MM. Relation between interatrial shunts and

decompression sickness in divers. Lancet 1989; II(8675): 1302-1306. Wilmshurst PT, Ellis BG, Jenkins BS. Paradoxical gas embolism in a scuba diver with an

atrial septal defect. Brit Med J 1986; 293(6557): 1277.

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Spatial and Seasonal Trends for Microbial Community Structure and Biomass across a Pollutant Gradient in the Western Basin of Lake Erie

J.A Porter* R.H. Findlay

Department of Microbiology Miami University

32 Pearson Hall Oxford, OH 45056

*Porterja@muohio.edu Abstract

The relationship between sedimentary microorganisms and lipophilic pollutants is complex in that many pollutants can serve as a resource at moderate concentrations but are toxic at higher concentrations. The western basin of Lake Erie has a natural pollutant gradient due to sedimentation patterns from the Detroit and Maumee Rivers with polychlorinated biphenyl (PCB) and polycyclic aromatic hydrocarbon (PAH) concentrations increasing northward toward Canada. Three stations were established along the gradient to elucidate the response of microbial communities to changing pollutant concentrations: low (PCB 36.83 ± 14.56 ng gdw-1, PAH 1.14 ± 0.39 µg gdw-1), intermediate (PCB 63.96 ± 4.06 ng gdw-1, PAH 1.62 ± 0.16 µg gdw-1) and high (PCB 135.23 ± 59.52 ng gdw-1, PAH 3.25 ± 0.60 µg gdw-1). Sediment samples were collected in May, July and September by SCUBA-assisted divers. Sediment samples collected in September were taken from within and between Dreissenid mussel beds (zebra and quagga mussels) to compare differences in microbial biomass and community structure as well as pollutant concentration. Sediment samples were extracted for microbial lipids, PCBs and PAHs using the modified, Bligh and Dyer lipid extraction. Microbial community structure was analyzed using phospholipid fatty acid analysis and pollutants by mass spectroscopy. Microbial biomass showed a trend of greater biomass at the low pollutant concentration station compared to the high pollution concentration station in May and July, with low and high pollutant stations having comparable microbial biomass in September. Microbial biomass was significantly higher between mussel beds compared to within beds at the low and high pollutant stations with no detectable difference at the intermediate pollutant station. Microbial community structure, analyzed using principal component analysis, showed a seasonal pattern with changes in dominance between phototrophic microeukaryotes and heterotrophic prokaryotes dependent on sampling date and independent of pollutant concentration. Stations showed significant differences in sediment surface areas; these changes did not correlate with microbial biomass, community structure or pollutant concentration. Once seasonal variation in community structure was removed, changes in microbial community structure correlated with pollutant concentration. Two marker fatty acids (i17:0 and a17:0; associated with gram-positive and anaerobic gram-negative bacteria) were enriched with increasing PCB and PAH concentration. These findings indicate that

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bacterial communities in Lake Erie sediments respond to pollutants and the presence of mussels within the sediments. This research is dependent upon SCUBA-based sediment collection techniques which facilitate the analysis of spatial affects on microbial communities in Lake Erie sediments.

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PRISM Topaz Mixed-Gas Closed Circuit System: Design and Development For Recreational, Scientific, and Commercial Users

Peter F. Readey Steam Machines Incorporated

Hermosa Beach, CA info@steammachines.com

Abstract

To effectively maximize underwater durations, many in the scientific, recreational and technical market are turning their attention to the field of closed circuit diving systems. Extended durations, reduced decompression obligations, and reduction/elimination of bubble exhaust contamination are just a few of the benefits offered by this mode of diving. The rebreathing concept pre-dates open circuit SCUBA, however, the material and physiological science at that time was not as advanced and this allowed simpler open circuit concepts and systems to predominate the diving market, despite their relative inefficiencies.

We will provide a brief overview of the different types of equipment and how such apparatus functions. We’ll discuss how technology has advanced to meet the rigorous demands of the underwater environment, including developments in scrubber design, sensors and electronics resulting in more efficient and reliable systems. Developments in physiology and a better understanding of how this equipment effects the diver has, through rigorous independent testing, lead to improved equipment designs, with emphasis placed on work of breathing effort, hydrostatic loading, oxygen control and ergonomics.

A review of training and logistical considerations is required for successful field applications of closed circuit systems, including an examination of project objectives. This critical element relies upon the educated diver matching appropriate units and finding an acceptable trade-off between system size, weight, and complexity versus training requirements and in-field support and maintenance.

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Shipwrecks and Students: Discovering Maritime Heritage

Timothy Runyan Maritime Studies Program

East Carolina University Greenville, NC 27858

Runyant@mail.ecu.edu Abstract Graduate students from across the United State and abroad come each year to begin their studies at East Carolina University at the MA and PhD level. Most of them are drawn to the Program by the opportunity to do underwater research using scuba and other systems. The Program conducts an extensive array of field experiences. This includes a summer field school for first year students, that follows the completion of a AAUS scientific diver training program offered by the Office of Diving and Boating Safety. These field schools are held in a variety of locations ranging from North Carolina to the Great Lakes and the US Virgin Islands. The MA degree is offered in Maritime History and Nautical Archaeology. This two-year program began in 1981 and has produced graduates in the role of responsible positions in the maritime field throughout the United States and abroad. Many are employed in federal and state government, maritime museums, by contract archaeology firms and teaching. Doctoral students in the Coastal Resources Management Program concentrate in one of the four major areas of ecology, geosciences, social sciences and maritime studies, while completing secondary fields in two of the remaining three areas. These students may be involved in diving and underwater research but have a focus on cultural resources management. This presentation will discuss the resources necessary to maintain the program and illustrate the opportunities in underwater research.

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Safe Harbors for our Future: An Overview of Marine Protected Areas

Derek Smith* Kathy Ann Miller

University of Southern California 1 Big Fisherman Cove

Avalon, CA 90704 *Dereks@usc.edu

Abstract

In the late 1800s and early 1900s, recreational fishers, especially in southern California, began to realize that populations of their favorite gamefish were in decline. Since 1952, 53 marine refuges, sanctuaries and ecological preserves, with varying levels of protection, have been established, totaling 2.2% of state waters. Of these, 10 are no-take reserves (0.2 % of state waters). In 1999, Californians voted to give the California Department of Fish & Game the responsibility for establishing a network of marine protected areas (MPAs) throughout the state. These portions of the coastline are set aside to protect and restore habitats, to conserve biological diversity, to serve as scientific baselines from which to judge future change in ocean ecosystems, to promote recreational and educational activities, and to enhance depleted fisheries. The science and politics of MPAs are hotly contested topics today. Although scientists agree that MPAs are important conservation and management tools, some stakeholders who exploit nearshore marine species for a living or for pleasure are not willing to set aside no-take reserves. The debate over their fairness to these stakeholders and also the efficacy of MPAs can derail or delay their establishment. Similarly, the site location and size of new MPAs depends not only on hydrographic and ecological criteria (the goal is to link reserves via larval dispersal) but also political and socioeconomic considerations. Once MPAs are established, their effect on nearshore marine community structure and on individual species population dynamics, compared to unprotected control sites, must be assessed by a comprehensive yet pragmatic monitoring program. A group of government and university biologists (CRANE) are currently developing state-wide protocols for subtidal monitoring using SCUBA and ROV surveys. Finally, adaptive management of MPAs and conscientious enforcement efforts are essential to the success of the reserve. Now is the time for intensive research to support the establishment of MPAs and demonstrate their efficacy for marine conservation.

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Diving on the Queen Anne’s Revenge

Chris Southerly* Jelep Gillman-Bryan

North Carolina Underwater Archaeology Branch 1608 Fort Fisher Blvd. South

P.O. Box 58 Kure Beach, NC 28449

*Chris.southerly@ncmail.net Abstract

First located in November 1996, the shipwreck of the Queen Anne's Revenge, the pirate Blackbeard's flagship, has become an international interdisciplinary research project. Archaeologists, scientists, and historians have collaborated under the direction of the North Carolina Underwater Archaeology Branch to analyze the shipwreck: identifying the extent of the site, assessing the condition of the cultural remains, studying the historical and modern environmental conditions of the area, evaluating potential impacts to site preservation, researching historical documentation, and collecting archaeological data.

Diving operations were an integral part of accomplishing or initiating much of the scientific research. Divers using open circuit SCUBA and wireless communication conducted detailed mapping, photography, videography, excavation, artifact recovery, and remote sensing.

A major educational initiative that was an overwhelming success was the QAR DiveLive. a distance education event primarily made possible thought the professional expertise of Nautilus Productions and Marine Grafics. For one week during both the fall 2000 and the fall 2001 field expeditions, live underwater video was transmitted to a shore facility that digitized the signal and uploaded it to the Internet.

While any Internet user could watch and listen to the divers at work underwater, registered school groups and museums could email questions that were relayed from shore to the site to be answered live by the support team on board the research vessel and/or the divers working underwater. DiveLive reached thousands of school children and countless viewers across the country and around the world, allowing them to participate firsthand in scientific diving as part of an underwater archaeological investigation.

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Use of Scientific Diving in the Coastal Ocean Research Monitoring Program (CORMP): Onslow Bay, North Carolina

Jason Souza* David Wells

University of North Carolina at Wilmington 5600 Marvin K. Moss Lane

Wilmington, NC 28409 *SouzaJ@uncwil.edu

Abstract

The Coastal Ocean Research Monitoring Program (CORMP) is a NOAA-funded research initiative aimed at better understanding the physical, chemical, and geological processes which shape the nearshore and offshore coastal ocean environment of Onslow Bay, North Carolina. Currently, CORMP consists of seven mooring sites (OB1-4, LB5-6, OB27). Each site is fitted with a bottom-mounted ADCP (acoustic doppler current profiler) and CT (conductivity/temperature) logger and mid-water CT loggers mounted on a taut-wire mooring which are diver-serviced every 3 months. The primary research site (OB27) employs additional instrumentation including a SCUFA and PC-ADP (pulse-coherent acoustic doppler profiler), which are all mounted on a stationary quadpod. Instruments at the OB27 site are diver-serviced every five to six weeks. Divers also take a number of different sediment samples for biological and geological research. Due to the nature of the project, CORMP investigators rely heavily on scientific diving. The varied scientific objectives coupled with the depths (60-140 FSW) and seasonal fluctuations in water temperature (40-82o F) require divers to utilize various techniques in completing their missions. We give a detailed summary of the scientific diving methods employed in the various aspects of the CORMP project.

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Underwater Crime Scene Investigations (UCSI), a New Paradigm

Gregg Stanton Florida State University at Panama City

4750 Collegiate Drive Panama City, FL 32405

Gstanton@pc.fsu.edu Abstract

A new paradigm in forensic investigation was recently launched in the School of Criminology and Criminal Justice at Florida State University in Panama City. Driven by the USS Cole terrorist attack and all events surrounding 9/11, a team of specialists from multiple disciplines such as underwater archeology, ocean engineering, mechanical engineering, marine biology, forensic science, and criminology was assembled to change underwater criminal investigation from a “snatch and grab mentality” to the rigors of a scientific study. The mission of this research "Alpha" Team was to identify and/or develop underwater criminal investigative protocols that follow national and international law. The Alpha Team was to evaluate, adapt and develop technology to quickly identify the suspects for a quick response. Once the protocols were developed, the Alpha Team members were to replicate themselves within a second or Beta Team structure. Funded by the Department of Defense and the Defense Threat Reduction Agency in support of a FEMA exercise, the Alpha Team instructors will ultimately create training modules for military and civilian law enforcement agencies, which will then carry the distinction of a Beta Team certification.

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New Insights into Sea Urchin Recruitment in the Gulf of Maine

Rebecca L Toppin* Larry G. Harris

Department of Zoology University of New Hampshire

46 College Rd. Durham, NH 03824

*Becca.toppin@unh.edu Abstract

Commercial harvesting of sea urchins has become a major industry in the Gulf of Maine following a population explosion of. Strongylocentrotus droebachiensis in the late 1970s. Recruitment studies begun in 1983 have continues through 2002. Recruitment of the green sea urchin, S. droebachiensis in the Gulf of Maine has decreased dramatically in recent years. New observations show that recruitment in the northern Gulf of Maine has remained low but predictable, while the decline in recruitment observed in the late 1990s at southern sites appears to have stabilized. Recently it has been noted that recruitment and survival are possible in the absence of adult S. droebachiensis. There have been observations of heavy recruitment of juvenile urchins into an algal dominated site with no adult urchins present. These findings are contrary to most previous studies that suggest the disturbance and spine canopy provided by adult urchins facilitates successful juvenile recruitment.

Growth studies in benthic and suspended cages showed differences in growth and mortality. Growth studies done with juvenile, hatchery-reared urchins showed large amounts of recruitment. Protection from potential predators was due to mesh size and suspension of the spat bags in the water column. Temperature may also play a role in sea urchin recruitment, growth and survival.

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To Catch a Flounder: the Potential for Escapement of Older Southern Flounder (Paralichthys lethostigma) from Fishing Pressure in North Carolina

Carter Watterson* John Alexander

North Carolina Division of Marine Fisheries 3441 Arendell Street, P.O. Box 769

Morehead City, NC 28557 *Carter.Watterson@ncmail.net

Abstract

Southern flounder (Paralichthys lethostigma) is the most economically important finfish species in North Carolina, USA. Landings for this species have averaged approximately 1,738 metric tons over the past eight years. The majority of the landings (93%) are comprised of one and two year-old fish. Considering the maximum age for the species is eight, this indicates either a truncated age structure for the population or escapement of the older fish from fishing pressure.

In North Carolina, southern flounder are highly susceptible to fishing pressure while inshore during the warmer months, primarily from gill nets and pound nets. As the temperature drops, the flounder move offshore into ocean waters to spawn. Once outside the inlets, very little exploitation of the stock occurs. It has been hypothesized that a portion of the population of older fish may remain offshore following spawning rather than returning inshore to the rivers and estuaries. To test this hypothesis, southern flounder have been collected on the natural and artificial reefs off of North Carolina throughout the year for aging. This presentation will focus primarily on the methodology surrounding the collection of the flounder in its natural habitat through the use of SCUBA, along with the inherent advantages and disadvantages to this collection method.

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What does a Sponge Eat? Examining Variability in Sponge Nutrition in the Florida Keys

Jeremy B. Weisz1* Melissa Southwell2

Christopher S. Martens2

Niels Lindquist1

Ute Hentschel3

1Institute of Marine Sciences University of North Carolina at Chapel Hill

3431 Arendell St Morehead City, NC 28557

*weisz@unc.edu

2Department of Marine Sciences University of North Carolina at Chapel Hill

CB# 3300 Chapel Hill, NC 27599

3Institut für Molekulare Infektionsbiologie

Universität Würzburg 97070 Würzburg, Germany

Abstract As filter-feeding organisms, sponges are intimately tied to their environment. Any variability in the environment should therefore be reflected in the tissue of the sponge. However, some sponges are known to host large microbial communities, and the influence of these microbes on the nutrition of their host is largely unknown. To examine the influences on variability in sponge nutrition, we collected tissue samples from eleven sponge species at thirteen ocean and bayside sites near Key Largo, Florida. We also collected sediment samples, suspended particulate organic matter, and seagrass samples where possible. All of these samples were then analyzed to determine the stable carbon and nitrogen isotope ratios, which are used as a tracer of nutrient flows between organisms. These data revealed significant inter- and intraspecific differences, such as low δ15N values that suggest N2 fixation occurs within some species, and spatially variable δ13C values that suggest variations in organic matter inputs between nearshore and offshore sites. To explore the potential connections between low δ15N values and the N2-fixing potential of sponge-associated microbes, and spatial correlations between δ13C values and variable sponge microbial communities, we performed molecular analyses on sponge tissue and water column samples to characterize the microbial communities of these samples. We also used fluorescein dye to trace the flow of water through the sponges, and were then able to collect excurrent water, which will also be analyzed to

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characterize the microbial community. These analyses indicate a correlation between variability in the stable isotopes and variability in the microbial communities. These data, therefore, suggest inherent differences in types and sources of nutrients available to sponge species that contain diverse and massive bacterial communities versus those with few bacteria. Future work includes a mission in Aquarius, the underwater habitat, to perform tracer experiments with sponges to determine rates of nitrogen fixation.

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The Introduction and Dispersal of the Indo-Pacific Lionfish (Pterois volitans) Along the Atlantic Coast of North America

Paula Whitfield1* Todd Gardner2

Stephen P. Vives3

Matthew R. Gilligan4

Walter R. Courtney Jr. 5G. Carleton Ray6

Jonathan A. Hare1

1National Oceanographic and Atmospheric Administration

101 Pivers Island Rd. Beaufort, NC 28516

*Paula.Whitfield@NOAA.gov

2Biology Department 130 Gittleson Hall Hofstra University

Hempstead, NY 11549-1140

3Department of Biology Georgia Southern University

Stateboro, GA 30460

4Marine Science Programs Savannah State University

P.O. Box 20467

5Florida Caribbean Science Center U.S. Geological Survey

7920 NW 71st St. Gainesville, FL 32653

6Department of Environmental Sciences

University of Virginia Charlottesville, VA 22904

Abstract

Pterois volitans is the first Pacific marine fish to become established in Atlantic waters. For the past 2.5 years, the Indo-Pacific species of lionfish have been collected, photographed and observed from southern Florida to Cape Hatteras, NC and juvenile lionfish have been collected from Long Island, NY and Bermuda. The large scale

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distribution of this invasive species over such a short time period (2000-2002), along with the presence of juveniles, suggests that lionfish are reproducing. Along the North Carolina coast both the number and spatial distribution of lionfish have increased. In August 2000, there were 3 lionfish reported in 3 locations. By October 2002, 49 lionfish were reported in 15 different locations. Dispersal of lionfish in 2002 appears to be both inshore and offshore of the lionfish locations in 2001. Lionfish have also been sighted in waters deeper than originally anticipated. The initial introduction was probably through aquarium releases off the Florida coast with dispersal of eggs and larvae through Gulf Stream transport. It is unknown what the ecosystem effects of this marine invasive fish will be along the Atlantic coast of the United States. There is cause for concern as they are predators and appear to be permanent residents.

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