Coralmania by Eric Borneman

Coral Culture for Disease Research and Reef Restoration


Many of you have written or spoken to me about the presentation I gave recently at the International Marine Aquarium Conference (IMAC 2004) in Chicago last June. Several of you requested a summary of what I spoke about, so I am providing a manuscript submitted to the Coral Disease and Health Consortium that documents our progress in the culture of Caribbean species, that fully explains the subject of my IMAC 2004 lecture.

To use skills learned as an aquarist to benefit science and coral reef conservation has been a long-time goal of mine as an aquarist. I have found myself in a somewhat special position, drawing on decades of experience as a diver, well over a decade of experience with the husbandry of corals in reef aquaria, and now a few amazing years as a coral reef scientist. It is equally special how these fields overlap and complement each other, but unfortunately few participants in each field are aware of the intricacies and specialized knowledge of the other disciplines. I think this is changing, as more and more scientists come in contact with successful coral displays, and as more and more aquarists are themselves scientists in various disciplines. Diving? Well, diving puts everything into perspective!

As aquarists, we have experienced around 20 years of success with keeping some corals alive, 10-15 years of success keeping almost all zooxanthellate corals alive, and only limited success with most of the azooxanthellate species. Certainly, the prolific growth of Aiptasia sp. anemones gave some clue that "farming" cnidarians was possible. In 1999, I wrote a paper (Borneman and Lowrie 1999) for which a friend of mine, Deborah Lang, did some serious legwork in helping me determine all the species of corals that were being propagated at that time (with some obviously questionable species that are difficult to identify). We were able to list some 400 species as available, although most were Acropora species and soft corals. Over the years, cultured corals have been made available to the hobby from mariculture operations, aquaculture facilities, and local sources (aquarium clubs, local trading, etc.). Entire businesses are now devoted solely to the propagation of corals, books have been written on the subject, and now some websites even offer "propagation tools," such as forceps, snippers, glues and mounting materials. Frags.org, a relatively new website and concept, is a loosely formed organization of some 250+ private individuals and businesses who offer cultured corals for sale or trade. Organizations like this are truly remarkable, having stemmed from earlier efforts such as the "League of Coral Farmers." Some coral farms, such as Tropicorium, Inc. have been around longer than many aquarists have kept corals, and the pioneering propagation work by Dick Perrin (and son) are models of the industry's potential.

Coral farming has other beneficial or economically valuable uses beyond the aquarium trade (Table 1). I have also compiled a table of sources of aquacultured corals (Table 2). I apologize to any people or companies who aquaculture but are not listed, and would encourage everyone I have omitted to contact me so that I can add their names to the list. While in most cases cultured coral is ecologically favorable to wild-collected coral, it is generally acknowledged that the economically preferable option is to allow coral collectors to maintain their employment by teaching them to become coral mariculturists, rather than supplanting resource country opportunities by displacing wild collection with land-based aquaculture outside those resource countries. The numerous attempts to begin coral mariculture have not, unfortunately, lessened the impact of wild collection to date. However, more and more mariculture operations are now forming, becoming viable, and substituting larger proportions of cultured corals for wild harvest. I wrote about such a coral farm, that was attempting to bring Caribbean cultured corals to the aquarium trade, several years ago.

Some aquarists have rightfully noted that maricultured corals are not as "hardy" as those grown in tanks. They are often just as demanding in terms of acclimation as wild colonies, and tend to suffer higher mortality than tank-cultured colonies. But, mariculture facilities are able to selectively culture the "best of the best" under conditions that, while still subject to environmental stochasticity (random variability), are far less variable than those affecting collections taking place over vast regions and very different habitats. Culture stock is selected for desirable traits such as color, shape, growth, and survivability. Much less selection is possible with wild harvest. Furthermore, maricultured corals are grown near the coast and near logistical transport, likely decreasing transport- and shipping-related stress and mortality. Mariculture facilities now exist or are planned in many resource countries (Table 3).

Outside the aquarium trade, managers, government, conservation-minded groups and scientists are also beginning to take note of coral farming's potential uses for various purposes. Several recent publications highlight and acknowledge the desirability of coral farming. A passage from the U.S. Coral Reef Task Force's, "Proposed National Action Strategy: Proposed Actions and Strategies to Address Key Threats states:

"The following actions are offered to address concerns:
Expand and strengthen capacity building efforts in countries with coral reefs to enforce relevant laws and regulations, collect trade data, and develop and implement sustainable management plans, certification schemes, and alternative harvest practices such as aquaculture and coral farming."

The use of novel methods of restoration, rehabilitation and replenishment using propagated corals has also been mentioned in statements from the International Coral Reef Initiative, the Proceedings of the Caribbean Acroporid Workshop, and numerous other publications. I have recently reviewed several articles that directly relate to the aquarium-based culture of corals for restoration, and other articles have appeared sporadically throughout the years, many of them by well-known professional aquarists of public aquaria (most notably, Walter Adey and staff at the Smithsonian Institution, Jean Jaubert and scientists at the Monaco aquarium and related facilities, and Bruce Carlson and staff at the Waikiki aquarium). The idea that growing corals is possible is still, however, a somewhat novel idea to many in the scientific community. I recently submitted a grant application to supplement funding for a project involving coral culture, and received a comment from a reviewer who remarked, "While this proposal is a novel idea, I do not think it is feasible or even possible to grow Acropora in closed system aquaria." Clearly, there is still a wide gap in the sharing of information between many scientists and aquarists.

Despite such comments, there is clearly a strong trend in science to begin approaching, experimenting with and utilizing various techniques and forms of coral culture to benefit research and reef restoration. Why is there a sudden interest? There are two major reasons. The first is that coral reefs are dying or degrading rapidly on a global scale. Notably, the near extirpation of the primary hermatypic Caribbean species, Acropora cervicornis and A. palmata, has created a desire for new ways to re-establish the species throughout their range. The second reason is the number of reefs that are candidates for reef replenishment or restoration due to ship groundings and other environmental impacts.

Reef restoration is big business and can involve large remediation sums and settlements, and private corporations offer various technologies to help restoration efforts. The use of artificial substrates, such as Reef-Balls™, and technologies such as mineral accretion, are often employed. The downsides to the use of many current restoration techniques are numerous.

  • They are both costly and not usually cost-effective.

  • They tend to be very labor intensive,

  • They are questionably effective (high mortality or low diversity/abundance of restored fragments), and

  • They are complicated.

Many restorations also involve a "borrow from Peter to pay Paul" ideology, whereby corals are removed from a donor site and replaced in a receiver site. When forests are restored from fire or logging, trees are not uprooted from other forests and taken to the damaged areas. Rather, small seedlings are planted from existing nurseries. Similarly, groups in Israel, Fiji, the Solomon Islands, Japan, and other places currently have coral nurseries in place using varying techniques, many of which are products of aquarium propagation methods. It is very encouraging to see these efforts taking form and being utilized. Coral nurseries, albeit ex situ (or aquaculture) rather than in situ (mariculture), are also where my role in this article comes about.

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An oft-stated ideal among reefkeepers is that, as aquarists, we can grow corals and help save reefs by putting them back out on the reef. The fact is that this is currently not, and may never be, a workable option. Any coral that has been placed into a reef aquarium has undoubtedly come into contact with many species that are not native to any putative area of reintroduction. In mixed displays containing Pacific and Caribbean species, this is even more of an issue since little mixing of benthic species occurs between the coral reefs of the tropical Atlantic and Indo-Pacific regions. Genome mixing by plasmid transfer is possible and even likely; such chimeral species with genes from several potential sources should not be released. Well-meaning aquarists often forget that although Free Willy was a great and inspiring movie, "releasing" clownfish, lionfish, corals, algae, and (more importantly) possibly invasive species or pathogens (likely invisible to the aquarist) is a very real danger and a very bad idea. Strict controls must be maintained to even consider such "home projects" as a viable option, and even then permission for reintroduction to any wild community would not likely be given.

To that end, an effort is now underway to develop a coral health certification whereby corals in culture would be designated as "healthy" and posing minimal risk for reintroduction or for transfer to other facilities. Certification would apply only to the target projects, but would also have many implications for other aspects of aquaculture, mariculture, and even wild harvest. Perhaps certification could even eventually be used to ensure healthy corals within the scheme of the export to retail aquarium trade chain. It would certainly be of great interest and benefit to the aquarium trade to know that corals offered for sale are known to be healthy through a systematic checklist of factors. I worked on the development of these guidelines with a large committee of experts in July 2004. Goals of the certification include: developing a best practice management for aquacultured corals from collection to quarantine and culture conditions; assessments of coral health throughout the processes; and the possible pretreatment/surface sterilization of colonies prior to reintroduction. These goals will help ensure that only healthy corals are returned to reefs. When completed, these guidelines will establish the first health standards for corals.

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While the major focus of our efforts is for the culture of clonal fragments for coral disease research, we have received funding and permission for test reintroductions to a ship grounding site in the Florida Keys. This study will compare the growth and survival of corals removed from the Truman Annex (see article below), and then grown in our facility, other aquaculture facilities in Florida, and a live rock mariculture site. The corals will then be explanted onto the test restoration site. Questions to be answered from this work are numerous. Is aquaculture a feasible way to generate coral fragments for restoration and/or rehabilitation? Can tank-raised corals compete and survive on the reef? Will tank-raised corals act as a vector for disease to wild communities?

Together with the owners of the coral farm discussed below, we also have plans for, or are involved in expansions of this work for other areas of research and conservation activities. All in all, this project is truly groundbreaking in many ways and has thus far been extraordinarily successful. What gives me great hope and pride is the enthusiastic reception from outside parties with nearly constant inquiries as to the further development of the project. We have what I believe to be the largest inventory of Caribbean corals in the country, and we do not plan to stop anytime soon. It is our goal to continue expanding our diversity and inventory to other areas and other species until we have a carefully organized, genetically diverse and characterized stock of corals available for use by all interested parties. Included in our plans are techniques to settle sexually produced coral larvae from spawning events to rapidly and sustainably increase production of coral fragments on a largescale. I attempted to induce settlement of larvae resulting from spawning events from the Flower Gardens in 2003, and my effort was a dismal failure and an obvious learning event. Now, I have contacted those who have utilized spawn settlement techniques to great success in Japan and Germany, and plan to try again this year during spawning events. Additionally, we will be receiving later this year a large quantity of aquarium-settled and grown colonies from Rotterdam as part of a joint effort to increase the use of sexually spawned and settled corals for various purposes (Dirk Peterson, pers. comm.).

Many of you have asked whether or not any of the Caribbean corals will be available to the aquarium trade. This answer is, we have no such plans at present. The "black market" corals I acquired from local Houston stores could technically be propagated and sold. The other collections are under strict permit requirements and can be used only for reef research and the goals outlined by the CDHC. Eventually, I hope that propagules from original broodstock or future acquisitions will be available, but we have no plans to approach this subject in the near future. The coral farm, Reef Savers Inc., will, however, be offering Pacific species on a large scale to the trade as broodstock is developed.

Currently in progress, we are continuing fragmentation of the many colonies that must be managed to maximize growth from the margins and produce appropriately sized specimens. We are beginning procedures to measure both linear growth and weight of corals to determine calcification rates. Because of the scale of the project, we are developing practical, logistical genotype separations so that we can track parent and resultant daughter colony propagules. We are setting up more systems, including special quarantine systems required in other large acquisitions, since several hundred or thousand pounds of coral entering a facility at any one time can be constrained by the extraordinarily large space requirements necessary to ensure protocol adherence; a situation not usually experienced by traditional aquarium trade acquisitions even by large wholesale facilities. Cost- and function-effective bases and mounting techniques are required for end-use purposes. We are also working on "microfragmentation" to increase the colonies' growth potential and to meet the very small size requirements for typical molecular research needs. The "fine-tuning" for large-scale water flow, plankton, calcification, and lighting needs of these species are constantly being "tweaked" to ensure the best progress of our efforts. Finally, we are building Acropora palmata- specific systems for future acquisitions of this highly demanding, but critically important, species.

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In terms of our current and future needs, we would like to dramatically expand our production of the Caribbean acroporids because of their ecological importance and the potential for their culture. The health certification must be developed and put into place. The rapid progress and unexpected success of the project, to date, have produced a somewhat more urgent need to develop formal arrangements for supply and demand within the CDHC plan, and to establish logistics for specimen transfer. While the corals produced for research will be sold in the same manner as "lab rats" provided by biological supply sources, the costs have not yet been determined because of the novelty of culturing Caribbean species on a large scale, and the break-even points to become a self-sustaining venture are still unknown. Efforts are underway to establish a non-profit division of Reef Savers that will be responsible for all research and conservation-related activities. This will, in turn, give us better access to other grant-funded opportunities to cover further development costs of the facility in current and future projects. Efforts to obtain supplemental funding support are currently underway.

Future plans of this project include cooperative work with researchers and managers for specific research, restoration and rehabilitation needs, including those outside the goals of the CDHC. We are working with interested parties and a new aquarium-trade conservation initiative, Reef Protection, International, to develop educational and outreach materials. We plan to work with other labs and facilities to develop redundant and specific systems for other interested parties and ensure widespread dispersal of clonal libraries and subsequent maintenance of existing genotypes. With future acquisitions, we will establish wider Caribbean location-specific systems and genotypes so that separation or mixing of genotypes for location-specific needs can be achieved. We will begin environmental tolerance tests and attempt to develop "high-tolerance" strains using corals' natural acclimatization abilities, focusing on environmental tolerances to the most common stressors facing natural communities, such as temperature and increased nutrient levels. Finally, we will work with all appropriate parties toward the formation of a coral "gene bank" that will require partial or total sequencing of genotypes in culture. Together, these efforts should be able to help move coral reef science forward to an appreciable extent, and it has been a great pleasure and honor to have been given such an opportunity - one that has exceeded our wildest expectations in a very short period of time. I also thank all of you who have subsequently offered products, financial support, or advice toward this project subsequent to my presentation at IMAC, 2004.

Acknowledgements

I would like to thank all of the following people for their contributions and help in this project: the Florida Keys National Marine Sanctuary, and especially Laurie McLaughlin, Joanne Delaney, Billy Causey, Harold Hudson, and Bill Keller; the Coral Disease and Health Consortium, and especially Cheryl Woodley, Esther Peters, Cindy Hunter, Andy Bruckner, Pam Parnell, Roy Yanong, and Ilze Berzins; the Tropical Aquaculture Center, especially Craig Watson; Ken Nedimyer; the Flower Gardens National Marine Sanctuary, and especially Emma Hickerson and G.P. Schmahl; Walt Smith; Dirk Peterson; Drew Weiner; the Florida Aquarium; Mote Marine Laboratory; Dennis Tagrin; Roger Vitko (Tunze USA); and especially my collaborative "partners," Eric and Kim Koch of Reef Savers, Inc.

Table 1. Potential uses of cultured corals:
Medical research (bone grafts, pharmaceuticals)
Coral reef research
Restoration
Reef replenishment
Aquarium trade
Jewelry
Curios
Fisheries
Coastal management
Tourism and diving
Employment in coral mariculture or aquaculture field

Table 2. Sources of cultured corals:
Frags.org - coral fragment trading group of individuals and businesses
Getting Tanked
Blowfish Aquatics
Reef Savers
Coral Planet
CoralSandBar
Gooch's Corals
Captive Reef Corals
Live Aquaria
Invert-ual Realities
eCorals
Tropical Paradise
Tampa Bay Saltwater
Reefdweller/Reefaholics
Harbor Aquatics
CoralFragz
The Coral Reef
Coral Reef Productions
AquariumCity
Atlantis Aquatics
Inland Aquatics
AquaTouch Livestock
Coral Dynamics
REEFlections
Premium Aquatics
Reefer Madness
Indo-Pacific Sea Farms
Splash of Life
Tropicorium
Dr. Mac & Sons Corals
Joe Kelley
Reeffarmers
The Reef Web
Rod's Reef
Marine Ecosystems
Captive Raised Corals
Inland Reef Aquaria
Midland Marine
GARF
Greg Hiller
Dynamic Ecomorphology
Salt Water Connections
Coral Reef Aquarium
Coral Connection
J & B's Marine Propagations
SeaCrop
Mermaid's Reef
TheSea.org
SeaCare
Reef Creations
Something Fishy
Captive Bred Corals
eTropicals.com
Lonestar Corals
Shawn & Michael Bennett's
Adrian's Coral Reef
Soutas Saltwater
AquaCorals
Monaghan's Corals
The Saltwater Reef
Ken's Reef
Amphibious
The Cultured Reef
South Pacific Aquatics
5th Day Aquatics
Bama Saltwater
The Frag Store
Tridacna Reef
Ocean Aquatics
Belau Aquaculture
Walt Smith International
CV Dinar
Blaine Perun's Farms at the Sea

Table 3. Countries with existing or
planned coral mariculture facilities:
Palau
Dominica
Australia
Puerto Rico
Tonga
USA (Florida)
Solomon Islands
Red Sea
American Samoa
Philippines
Marshall Islands
Indonesia
Fiji
Israel
Japan



Model Systems and Coral "Lab Rats" for the Coral Disease and Health Consortium: Large-Scale Culture of Reef Corals for Disease Research

Eric Borneman1, Eric Koch and Kim Koch2

1Department of Biology, University of Houston, Houston TX eborneman@uh.edu
2Reef Savers, Inc., Stafford, TX eric@reeftanks.org

Draft manuscript prepared for the CHDC Workshop on coral disease diagnostics. Madison, Wisconsin. April 27-29, 2004.

Introduction

This project implements one of the Coral Disease and Health Consortium's high priority recommendations: to identify and develop model coral species (analogous to "laboratory rats") that are well characterized and suitable for laboratory culture, and to establish a culture facility able to supply model species for scientific research.

To investigate normal coral biology and disease states using modern scientific techniques, it is necessary to identify and develop model species, and make them routinely available for research. Model laboratory species share well-known desirable characteristics, including ease of culture, high growth and fecundity rates, and relatively simple genetics. Model corals will be analogous to "lab rats," and enable rapid advances by focusing research on fundamental biological concepts broadly applicable across the taxa. Model corals must be representative of coral diversity, and include Indo-Pacific and Caribbean species, autotrophs and heterotrophs, branching and boulder growth forms, species with different calcification rates, and with different algal symbionts. They must be susceptible to bleaching and disease, and also include taxa that are resistant to bleaching and disease. Developing a living stock collection for model corals will provide infrastructure critical for basic research by providing well-characterized and documented experimental organisms to domestic and international researchers.

The Coral Disease and Health Consortium (CDHC) has determined coral culture of key species to be integral in furthering the study of coral disease. At the first CDHC workshop (Charleston, South Carolina 2001), the following were deemed essential among the numerous high priority action items: 1) to be able to keep corals alive for study in controllable conditions, and 2) to establish clonal lines and the equivalent of coral "lab rats" that can be used to establish baseline data. These items were deemed imperative in the study of coral disease in order to gain efficacious results to help ameliorate pathologies causing rapid and severe coral mortality and resultant ecosystem damage. Limitations in the abilities to provide research corals and to maintain colonies in captivity were seen as hampering research progress, and it was acknowledged that most animal and human research depended on the existence of such basic needs. The U.S. Coral Reef Task Force, the International Coral Reef Initiative, NURP/CRMC, and the NOAA Coral Conservation Program (in conjunction with the National Marine Fisheries Service) have all identified the needs of this project as one of several key items to be addressed as part of their national action plans designed for the conservation of and addressing the global decline of coral reefs, including efforts to reduce impacts from overfishing, pollution, habitat destruction, disease, coral bleaching, and damaged reefs. Additionally, among the proposed solutions are novel strategies to reduce these impacts to coral reefs, including the use of restoration and culture-based technologies, the development of new techniques and technologies for marine research, and the culture of organisms to develop new value from the sea through aquaculture.

The goals of maintaining coral cultures are attainable using widespread techniques that, unfortunately, have not been widely disseminated amongst the coral research community, and such abilities have largely and erroneously been thought to be the exclusive domain of a few unusual persons or facilities (Carlson 1999). To date, research culture attempts in aquaria have met with very limited success, but are thought to have potential (Becker and Mueller 1999). Mariculture efforts are hampered by the fact that conditions in the ocean are subject to many of the same conditions that wild colonies face, and disturbances can destroy such efforts entirely (Bak and Criens 1981, Sakai et al. 1989, Yap and Gomez 1985). In particular, laboratory or aquarium culture attempts of Caribbean Acroporids have been largely unsuccessful (see for example, Becker and Mueller 1999). However, those attempting to culture Acropora in the past have failed to meet the requirements of the species in culture facilities, or lacked the skills required to maintain and grow these decidedly sensitive species (Gaines, pers comm.). Under proper conditions, growth rates easily match or exceed those found in the wild, and organisms can be raised in controlled areas free of storm damage, predation, encroachment, disease, and other variables inherent to in situ study (Borneman and Lowrie 1999). These techniques have allowed the long-term maintenance, growth and reproduction of every zooxanthellate and hermatypic coral made available, encompassing most species of Scleractinia and including several hundred species of other taxa (Octocorallia, corallimorpharians, etc.).

Corals grown by asexual means are of the same genotype, a characteristic of great benefit in coral research. Corals grown in aquariums can be acclimated to changing conditions, and could be reintroduced to increase the diversity or number of damaged, declining, or impoverished reef areas. Finally, species grown in captivity have been overwhelmingly reported to be exceptionally durable, and may, in fact, be more likely to establish themselves and survive adverse conditions as they mature (Carlson 1999). The ability of corals grown in captivity to adapt to environmental variations outside their normal range is well known.

The objectives of the CHDC committee charged with developing the goals were to determine which species are most suitable for use as coral "lab rats." Requirements for species were decided as follows: able to be maintained in captivity; susceptible, resistant, or appropriate to conditions being studied; availability; preferably fast growth; existing database within literature or other sources; able to be propagated, sexually or asexually; ecological important species. For the Caribbean, the following species were chosen: Acropora cervicornis and A. palmata, Siderastrea siderea, Montastraea annularis/faveolata/franksi, Porites astreoides and P. porites, Agaricia agaracites, and Gorgonia ventalina.

Facility

The facility being used to develop clonal lines is Reef Savers, Inc., a large coral farm in Stafford, Texas. Several systems were set aside and designated exclusively for the project, with numerous systems available for expansion, if required. The systems are composed of 250-gallon acrylic tank modules, with a total system volume of approximately 5000 gallons. Each module tank shares the same system water volume, although each tank is capable of being isolated by shut off-valves from every other tank in the module. Each tank is illuminated by various combinations of metal halide lamps, ranging from 250 - 400 watts utilizing different spectrums from 6500K to 20,000K, depending on the depth and irradiance levels that are being emulated. Additionally, very high output actinic fluorescent lamps are used to simulate dawn and dusk and allow for a more gradual and natural increase and decrease in photosynthetic rates of coral colonies.

The tanks are designed to maximize gas exchange and ensure saturated or supersaturated levels of oxygen. Water flow is provided by a number of methods. Primary flow is accomplished large diameter outflows of commercial grade centrifugal pumps. By themselves, they provide unidirectional flow along the length of the tank that is between 5-20 cm/second depending on the distance from the pump outflow. This alone provides water flow equivalent to areas of maximal coral diversity. In addition, large outflow "propeller" powerheads (Tunze Stream pumps) provide turbulence against the laminar flow with total flow per powerhead of approximately 7,000 - 10,000 gallons/hour. Finally, surge tanks can be used in systems designed to emulate to high-energy environment, allowing an environment conducive to rapid growth of shallow water species.

Water quality is supplemented on each system by the use of large protein skimmers, each powered by a designated high-pressure commercial pump. A sand bed is used in each tank of fine-grained oolitic aragonite sand to provide habitat for decomposing meiofauna and microbial flora that is a primary source of nutrient processing. Additionally, live rock is used in the sump of each stack to increase habitat and biodiversity for meiofauna and the production of demersal zooplankton. Pacific systems utilize Pacific live rock and Caribbean systems utilize Atlantic aquacultured live rock. Water for the systems is artificial, utilizing Crystal Seas BioAssay formula (Marine Enterprises, Inc.). Source water is purified through a commercial mixed bed deionization and reverse osmosis system, and mixed seawater additionally filtered though a 2 micron cartridge filter to further remove impurities.

Calcium and carbonate is replenished by several means; through the addition of reagent grade calcium chloride and sodium carbonate, and by custom-made calcium reactors. The reactors operate by bubbling carbon dioxide through a specially designed chamber containing various natural marine sources of calcium carbonate that we have mixed to maximize various minor and trace element constituents. No other elements are added as supplements to the systems.

There are many other very special and unique aspects of design and husbandry involved at this facility, but which are beyond the scope of this article.

Coral Collections

The collection of corals for this project include both Caribbean and Pacific species. Pacific species are acquired from a number of sources, including Fiji, Indonesia, Tonga, and the Solomon Islands. Additionally, a number of previously cultured genotypes have been acquired from various sources and their origin is unknown. Although the current livestock is limited to the species presented in Table 2, the acquisition of additional species is ongoing and can include virtually any species that is requested by the research community. Access to most tropical Indo-Pacific species is almost limitless and corals are easy to acquire. The majority of the information below is in regard to the Caribbean species since the Indo-Pacific species are so readily available. Their culture encompasses the majority of the facility and it is almost "second nature" for us to grow and propagate these species.

Caribbean species were initially slow to acquire. A limited sampling of a few colonies was permitted at the Flower Gardens National Marine Sanctuary. This provided an initial small broodstock of Porites astreoides, P. divaricata, and Montastraea faveolata. Attempts were made to acquire Acropora from Belize, but permitting constraints on the amount of material made the effort unfeasible. In November 2003, the Florida Keys National Marine Sanctuary made available many corals from the Truman Annex in Key West where a Navy dredging effort for the expansion of the port threatened a seawall covered by corals. These colonies were removed for relocation and many were provided, in part, for the CDHC culture systems. In January 2004, a second collection effort was made at the Truman Annex site and involved the acquisition of several hundred colonies. Small colonies of Acropora cervicornis that had recruited onto a live rock aquaculture lease site were provided by Ken Nedimyer (Sea Life, Inc.).

In March 2004, we received word that numerous Caribbean corals had been sold to several aquarium stores in the Houston area, likely the result of black market sales by an unscrupulous collector. The corals were immediately collected from the stores and, after two weeks of quarantine and surface sterilization using Lugol's solution and gentamycin, were also placed into the culture system. These colonies were reportedly obtained in the Bahamas. Finally, from February to April, 2004, several species were sent to us from aquarists who had obtained them as settlements on aquacultured live rock (Tampa Bay Saltwater) and subsequently propagated them by fragmentation. These corals were also subjected to the quarantine and sterilization procedure. Future plans will involve collections from Puerto Rico, Panama, and Dominica. Long-term plans include increasing the genetic diversity through acquisitions from throughout the Greater Caribbean region.

Coral Acclimation and Fragmentation

While some of the colonies acquired have been small (< 10cm), some colonies from the Truman Annex site were quite large (>50cm). Initially, stabilization of the colonies was essential, and they were placed immediately into the systems after removal of potentially fouling organisms (tunicates, sponges, and large iron deposits on the underside of the skeleton). Some colonies were flushed well with seawater to remove excess mucus and limit the organic loading on the tank systems. Ozone was employed by injection into the protein skimmer for several days while the corals acclimated to tank conditions and lowered their stress responses (increased mucus production, some mesenterial filament extrusion). This also reduced organic loading from epibionts, some of which were in the process of dying as a result of the collection and transport.

Lighting was provided on a reduced photoperiod (six hours versus the normal 12 hour period), and the initial Truman Annex collection proved that even this level of irradiance was too much initially for these turbid water collections. Several colonies of Agaricia agaracites, Favia fragum, Diploria strigosa, and Montastraea faveolata displayed various degrees of partial bleaching. Upon reduction of irradiance using only the fluorescent lights, all but two F. fragum and one A. agaracites recovered quickly. At that time, the 250-watt metal halides (6500K) were started on an increasing photoperiod, starting at 2 hours per day and increasing by 2 hours every week thereafter until the full photoperiod was reached. At this point, some colonies were moved under 400 watt metal halide lights (6500K and 20,000K, depending on observations of their responses and the species) to increase growth rates. All subsequent collections have followed this general protocol, and no further bleaching has occurred except in several colonies of Agaricia humilis from the second Truman Annex collection. These corals were immediately shaded and they completely recovered their symbionts within a week.

Most colonies are placed on top of eggcrate that is raised above the substrate by a few inches using 2" PVC pipe as "legs." Larger colonies, irradiance sensitive species (Agaricia spp., F. fragum), shade-loving (Scolymia spp.), and free-living species (Manacina areolata) are placed either directly on the sand substrate or are placed on eggcrate that sits directly on the sand. The species are located close or far from water flow depending on the species requirements or tolerances. Genotypes are segregated, where known, as are collections from the various regions.

Following stabilization, and once growth over any broken margins had occurred, fragmentation has been initiated. We have used lapidary and tile saws fitted with very thin diamond lapidary blades, and using seawater in the wet saw reservoir, to fragment the massive species. Fragment sizes may vary, but around 2-5 cm fragments are an ideal size to encourage rapid spread from multiple margins. Branching species are fragmented with wire snippers. All cut pieces are rinsed thoroughly in seawater prior to returning them into the systems.

Because of the numbers of corals involved, we have only begun the fragmentation process to date. Many of the fragments still require attachment to a base, the design and production of which is still being developed. Some fragments have been attached to small pieces of live rock, and others to a molded base made of unsanded, uncolored tile grout (calcium carbonate) using calcium chloride to speed the curing and provide an added boost of calcium in the substrate to potentially encourage rapid calcification. The molds are fitted with an acrylic block in the center to create a square hole while the grout cures, and is then removed leaving an indentation for cut and trimmed fragments to be mounted with live tissue touching the edges of the cured grout. Any space around the fragment is filled with aragonite sand and then hardened using a few drops of thin-set cyanoacrylate. This method was developed for these fragments and is proving to be effective for the massive and plating species. Branching corals are affixed using either high-density cyanoacrylate or fast curing epoxy putty, depending on the size and shape of the fragment. They are also mounted sideways to encourage basal spread by allowing maximal healthy tissue contact with the substrate, and allowing the broken edge and injured tissue to be exposed to strong water flow that speeds recoverage of skeleton by tissue and rapid recovery by exposure to strong water flow.

Mortality and Growth

Of the several hundred original colonies, three small colonies have been lost from a failure to recover from bleaching induced by the systems' intense lighting (F. fragum [2], A. agaracites [1]). One colony (Colpophyllia natans) was lost from a "brown jelly" infection that occurred after damage to the colony during the transportation process. Approximately sixty small fragments of M. faveolata were lost after the second Truman Annex collection. The colonies had been previously collected and amassed in plastic bins that were placed underneath a dock in the harbor. The fragments were piled on top of each other, and the local conditions (low water flow, shading) as well as physical damage from abrasion resulted in very weak fragments prior to transport. There was also a white material surrounding and affecting many polyps on the fragments, and this condition spread to affect adjacent polyps. These fragments were temporarily housed in a separate tank for observation and recovery, if it was going to occur. The fragments continued to decline, and were dead within a week. These were never added to the main culture systems. Several fragments were removed and fixed prior to their loss for further analysis of the white material.

The other 630+ colonies from the Truman Annex site have absolutely thrived in the system. They appear to be healthier now than on the marginal habitat where they were originally located. All fragments are displaying extensive polyp expansion and reproduction, with growth occurring rapidly on all colonies. Several first "generation" fragments were ready for a second division after a period of only four months. Notably, the A. cervicornis colonies are thriving and growing rapidly, and all have formed new branches and many new axial corallites. As we thought, the husbandry of this species does not appear to be difficult at all when the proper techniques and skills are employed. Also surprising is the rapid growth of massive species, known to be slow-growing in the wild. It appears the slow growth is a result of their colony formation, with propagation that enhances marginal expansion allowing for rapid growth. The corals are being fed multiple times per day using various zooplankton substitutes, including newly hatched Artemia nauplii, Golden Pearls (Brine Shrimp Direct), and CyclopEze. Additional live culture systems that will provide a constant drip of prey items are being constructed to increase colony growth and health.

Future Plans

Special treatment protocols will be initiated on subsamples of clonal lines to enhance tolerance of numerous environmental stressors currently impacting the species habitats; data will be collected regarding the tolerance levels of the species to various environmental parameters. Findings will be communicated to the scientific community, management authorities, and the public through publications and workshops, with the culture techniques being taught to any interested parties as part of the CDHC goal of maintaining "coral guinea pigs" through no-impact culturing systems for study and research. The benefits of establishing a stock of these critically important species is an essential step in understanding the requirements of these species and the conservation of coral reefs.

One of the primary goals of future efforts is to acquire larger stock populations of the critically important A. cervicornis and A. palmata species. We are confident that large-scale propagation of A. cervicornis will be possible. This species is among the most important to Caribbean reefs, but because of its rarity today, is difficult to acquire and is carefully guarded by managers. We would urge resource managers to allow collections of this species to enable us to rapidly reproduce fragments for research (or rehabilitation efforts). We are equally confident that we can grow the notoriously sensitive and demanding A. palmata. We believe the key to successful culture of this species will be in the efficient and proper transport of collected fragments, and that once in systems specifically designed to meet the species requirements, they will thrive just as every other Acroporid does when proper husbandry is provided.

In summary, a continuous source of fragments for use in research, reef rehabilitation and restoration projects will soon be available, with each system allowing fragments to be available for each collection region. Additionally, clonal coral specimens will be available for research that do not impact the wild, and do not require time consuming or expensive collection trips to obtain. Any corals with signs of disease or other pathology, if it occurs while in culture, will be isolated immediately and available for study by interested research parties. Valuable data will be gained from the treatment acclimated corals and their ability to withstand continued and unmitigated stressors present in wild communities. The ability of corals to adapt to local conditions is critical in assessing management strategies and in forecasting the potential of reefs to recover from disturbances. Tolerances of all clonal cultures to a variety of environmental parameters will be quantified. This information will provide necessary data to address most of the unknown stressor-response characteristics of corals in the Caribbean.

The results of this project to date have been extraordinarily successful. We have, through the opportunities afforded by the Truman Annex collections, acquired broodstock of nearly all zooxanthellate hermatypic genera in the Caribbean. This far exceeds the original proposal of focusing on a few key species. It is anticipated that the success of this project will ultimately result in significant progress in disease research, reduction of the collection of wild colonies that impacts genotypic diversity and coral coverage on a reef, and the eventual return of large numbers of viable colonies to the reef. These colonies would then be able to contribute to future settlements of colonies through spawning and natural fragmentation, enhancing the function of the reef as habitat and increasing biodiversity at the site.

Literature Cited:

Bak, R. P. M., and S. R. Criens. 1981. Survival after fragmentation of colonies of Madracis
mirabilis, Acropora palmata
, and A. cervicornis (Scleractinia) and the subsequent impact of a coral disease. Proc 4th Int Coral Reef Sym 2: 221-7.

Becker, L. C., and E. Mueller. 1999. The culture, transplantation and storage of Montastraea
faveolata, Acropora cervicornis
, and Acropora palmata: what we have learned so far. Bull Mar Sci 69: 881-896.

Borneman, E. H., and J. Lowrie. 1999. Advances in captive husbandry and propagation: an
easily utilized reef replenishment means from the private sector. Bull Mar Sci 69: 897-914.

Carlson, B. A. 1999. Organism responses to rapid change: what aquaria tell us about nature.
American Zoologist 39: 44-55

Sakai, K., Nishihiri, M., Kakinuma, Y. and Song, J. 1989. A short-term field experiment on the effect of siltation on survival and growth of transplanted Pocillopora damicornis branchlets. Galaxea 8: 143 - 156.

Yap, W. and Gomex, E. 1985. Growth of Acropora pulchra . III. Preliminary observations on the effects of transplantation and sediment on the growth and survival of transplants. Marine Biology 87: 203 - 209.

Table 1. Physico-chemical parameters of Reef Savers, Inc.
Variable Range Average

pH

8.2 - 8.4

8.2

Alkalinity (meq/l)

3.5 4.5

4.2

Calcium (ppm)

375 450

420

Ammonia*

unmeasurable

unmeasurable

Nitrite*

unmeasurable

unmeasurable

Nitrate*

unmeasurable

unmeasurable

Phosphate*

unmeasurable

unmeasurable

Oxygen (% saturation)

89 (night) 116 (day)

102

Salinity (ppt)

35-37

35.5

Temperature (oC)

24-28

26.5

Irradiance (E/m2/s-1)

250  - 700

 

* The test kits used are low range colorimetric field kits without the power to resolve the low levels of nutrients in the systems.

Table 2.  Pacific Species in Culture, full production

Name

Number of colonies

Fragments possible

Pocillopora damicornis

200

5,000

Acropora spp. (many)

1,000

20,000

Stylophora pistillata

200

5,000

Totals

1,400

30,000


Table 3.  Atlantic Species in Culture, current production
Locations possible Name Number of colonies Fragments

KN

Acropora cervicornis

7

tbd*

TA, AC

Agaricia spp.

49

tbd*

TA

Colpophyllia natans

5

tbd*

TA, AS

Dichocoenia stokesi

14

tbd*

TA

Diploria strigosa

75

tbd*

TA

Diplora clivosa

1

tbd*

TA

Diploria labyrinthiformis

4

tbd*

TA

Eusmilia fastigiata

25

tbd*

TA

Favia fragum

11

tbd*

TA

Isophyllia sinuosa

4

tbd*

TA, AS

Madracis sp.

24

tbd*

AC

Manacina areolata

4

tbd*

TA, AS, AC

Millepora alcicornis

12

tbd*

TA, AS

Montastraea cavernosa

74

tbd*

TA, AS

Montastraea annularis

3

tbd*

TA, FG

Montastraea faveolata

92

tbd*

TA

Montastraea franksi

3

tbd*

TA

Mussa angulosa

1

tbd*

TA

Mycetophyllia sp.

8

tbd*

TA, AC, AS

Oculina diffusa

54

tbd*

TA

Oculina varicosa

1

tbd*

TA, FG

Porites astreoides

39

tbd*

TA, AS

Porites porites

12

tbd*

TA

Scolymia sp.

10

tbd*

TA

Siderastrea radians

9

tbd*

TA

Siderastrea siderea

21

tbd*

AS

Solenastrea hyades

1

tbd*

TA

Stephanocoenia micheleni

72

tbd*

Totals

 

635

> 7,000

KN = Ken Nedimyer  AS = Aquarium store  TA = Truman Annex  FG = Flower Gardens AC = Aquacultured
*tbd (to be determined): The fragmentation of whole colonies is not yet completed; however, a general estimate is given for stock on hand.

 




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Coral Culture for Disease Research and Reef Restoration by Eric Borneman - Reefkeeping.com